Novel aqueous chitosan-based dispersions as efficient drug delivery systems for topical use. Rheological, textural and release studies.

The use of a novel cross-linked thiolated chitosan (CTS) was investigated as the main component of aqueous dispersions (at 1% and 3% w/v) for topical drug delivery systems. The nonionic theophiline (Th) and the cationic diltiazem(.)HCl (Dt) (at 0.5% w/v concentration) were used as model drugs. All aqueous dispersions behaved as viscoelastic fluids. The CTS 1% dispersions showed predominance of viscous component and low viscosity. However, in the CTS 3% dispersions, both the elastic component and high viscosities prevailed. So, texture parameters improved from CTS 1% to 3% dispersions and CTS 3%-Dt showed greater cohesion and adhesion than CTS 3%-Th, but always below CTS alone. All dispersions showed a Fickian diffusion mechanism. Despite release profiles of both drugs almost fully overlapped at 1% CTS, diffusion coefficients confirmed Dt released faster than Th at 3% CTS. The rheological behavior and the chemical nature of the drugs explained these results.

A new biodegradable polythiourethane as controlled release matrix polymer.

The main aim of this paper is the synthesis and characterization of a new linear functional biodegradable polythiourethane-d,l-1,4-dithiothreitol-hexamethylene diisocyanate [PTU(DTT-HMDI)]. The SeDeM diagram has been obtained to investigate its suitability to be processed through a direct compression process. Furthermore, the ability of this polymer to act as controlled release matrix forming excipient has been studied. Four batches of matrices containing 10-40% of polymer and theophylline anhydrous as model drug have been manufactured. Release studies have been carried out using the paddle method and the polymer percolation threshold has been estimated. The principal parameters of the SeDeM Expert system, such as the parametric profile (mean radius) and the good compression index (IGC=4.59) for the polymer are very close to the values considered as adequate for direct compression even with no addition of flow agents. Furthermore, the results of the drug release studies show a high ability of the polymer to control the drug release. The excipient percolation threshold has been estimated between 20% and 30% w/w of polymer.

Stabilization of apoptotic cells: generation of zombie cells.

Apoptosis is characterized by degradation of cell components but plasma membrane remains intact. Apoptotic microtubule network (AMN) is organized during apoptosis forming a cortical structure beneath plasma membrane that maintains plasma membrane integrity. Apoptotic cells are also characterized by high reactive oxygen species (ROS) production that can be potentially harmful for the cell. The aim of this study was to develop a method that allows stabilizing apoptotic cells for diagnostic and therapeutic applications. By using a cocktail composed of taxol (a microtubule stabilizer), Zn(2+) (a caspase inhibitor) and coenzyme Q10 (a lipid antioxidant), we were able to stabilize H460 apoptotic cells in cell cultures for at least 72 h, preventing secondary necrosis. Stabilized apoptotic cells maintain many apoptotic cell characteristics such as the presence of apoptotic microtubules, plasma membrane integrity, low intracellular calcium levels and mitochondrial polarization. Apoptotic cell stabilization may open new avenues in apoptosis detection and therapy.

Synthesis and characterization of a novel chitosan-N-acetyl-homocysteine thiolactone polymer using MES buffer.

We report a new "green" approach to synthesize a novel thiolated chitosan conjugate, chitosan-N-acetyl-homocysteine thiolactone (chitosan-AcHcys) using a "Good's buffers", 2-(N-morpholino)ethanesulfonic acid (MES). After that, the crosslinked Xr-chitosan-AcHcys was obtained only in the presence of air, without other reactants. The chitosan-AcHcys spectrum shows a partial incorporation of the thiolactone onto the polymer backbone. The derivative thermogravimetric analysis confirmed that chitosan-AcHcys is slightly less stable than starting chitosan; however, the peak profile is broadened which is indicative of deeper changes in the thermal degradation process. Also, aqueous dispersions with different concentrations of the crosslinked material (Xr-chitosan-AcHcys) were prepared and rheologically characterized. All aqueous dispersions are viscoelastic fluid with shear-thinning behavior. The viscosity of the dispersions (1-7% of chitosan-AcHcys) increases as a function of polymer concentration. So, we have achieved to disperse a high concentration of thiolated-chitosan derivative in water with different rheological characteristics, which could affect the drug release.

Apoptotic microtubules delimit an active caspase free area in the cellular cortex during the execution phase of apoptosis.

Apoptotic microtubule network (AMN) is organized during apoptosis, forming a cortical structure beneath plasma membrane, which has an important role in preserving cell morphology and plasma membrane permeability. The aim of this study was to examine the role of AMN in maintaining plasma membrane integrity during the execution phase of apoptosis. We demonstrated in camptothecin-induced apoptosis in H460 cells that AMN delimits an active caspase free area beneath plasma membrane that permits the preservation of cellular cortex and transmembrane proteins. AMN depolymerization in apoptotic cells by a short exposure to colchicine allowed active caspases to reach the cellular cortex and cleave many key proteins involved in plasma membrane structural support, cell adhesion and ionic homeostasis. Cleavage of cellular cortex and plasma membrane proteins, such as α-spectrin, paxilin, focal adhesion kinase (FAK), E-cadherin and integrin subunit ß4 was associated with cell collapse and cell detachment. Otherwise, cleavage-mediated inactivation of calcium ATPase pump (PMCA-4) and Na(+)/Ca(2+) exchanger (NCX) involved in cell calcium extrusion resulted in calcium overload. Furthermore, cleavage of Na(+)/K(+) pump subunit ß was associated with altered sodium homeostasis. Cleavage of cell cortex and plasma membrane proteins in apoptotic cells after AMN depolymerization increased plasma permeability, ionic imbalance and bioenergetic collapse, leading apoptotic cells to secondary necrosis. The essential role of caspase-mediated cleavage in this process was demonstrated because the concomitant addition of colchicine that induces AMN depolymerization and the pan-caspase inhibitor z-VAD avoided the cleavage of cortical and plasma membrane proteins and prevented apoptotic cells to undergo secondary necrosis. Furthermore, the presence of AMN was also critical for proper phosphatidylserine externalization and apoptotic cell clearance by macrophages. These results indicate that AMN is essential to preserve an active caspase free area in the cellular cortex of apoptotic cells that allows plasma membrane integrity during the execution phase of apoptosis.

Variation in scale shape among alternative sympatric phenotypes of Arctic charr Salvelinus alpinus from two lakes in Scotland.

Landmark-based geometric morphometric analysis was used to detect differences in scale shape between ecologically distinct phenotypes of Arctic charr Salvelinus alpinus coexisting in the same lake. Relative warp analysis and standard multivariate analyses of the partial warps, obtained after a Procrustes superimposition, showed that scale landmarks were efficient in discriminating among two closely related alternative phenotypes within each of the two lakes. In Loch Tay, S. alpinus exhibited a bimodal body size-frequency distribution among sexually mature fish, whereas in Loch Awe, S. alpinus are unimodal in body size but segregated into two distinct spawning phenotypes. In both lakes, alternative phenotypes showed significant differences in foraging ecology, habitat use and life history. It is probable that differences in scale shape reflect differences in ecology of these forms.

[Secondary serologic responses to dengue epidemic in 1998 in Salta, Argentina, where other flavivirus co-circulate]. / Respuestas serológicas secundarias en la epidemia de dengue de 1998 en Salta, Argentina, donde co-circulan otros flavivirus.

In 1998, a dengue outbreak (serotype 2) occurred in Salta province in Northern Argentina, following the first detection of dengue in the same area in 1997. We classified the serologic response of cases from 1998 as primary or secondary, since the risk of severe disease is greater for secondary cases. We studied 154 cases by plaque reduction neutralization and hemagglutination inhibition tests. Thirty-eight cases (25%) were classified as primary serologic responses and 84 cases (54%) as secondary responses. Thirty-two cases (21%) with borderline IgG titers could not be classified. Previous exposure to potentially cross-reacting flaviviruses (Saint Louis Encephalitis [SLE] and Yellow Fever [YF] viruses) was analyzed, as a possible cause of the secondary response pattern. Our results indicated that among cases classified as dengue secondary response, 83% could be attributed to previous SLE or YF exposure or serologic cross-reactivity. Vaccination against YF virus was at most a minor contributor to the secondary response pattern. The finding of a positive YF serologic result among persons not vaccinated may indicate silent circulation of YF in a region that can support both urban and jungle cycles. Other cases showing dengue secondary responses remained unexplained, suggesting the unrecognized occurrence of a previous infection with other dengue serotypes or of flaviviruses other than SLE or YF.

Respuestas serológicas secundarias en la epidemia de dengue de 1998 en Salta, Argentina, donde co-circulan otros flavivirus. / [Secondary serologic responses to dengue epidemic in 1998 in Salta, Argentina, where other flavivirus co-circulate]

In 1998, a dengue outbreak (serotype 2) occurred in Salta province in Northern Argentina, following the first detection of dengue in the same area in 1997. We classified the serologic response of cases from 1998 as primary or secondary, since the risk of severe disease is greater for secondary cases. We studied 154 cases by plaque reduction neutralization and hemagglutination inhibition tests. Thirty-eight cases (25

) were classified as primary serologic responses and 84 cases (54

) as secondary responses. Thirty-two cases (21

) with borderline IgG titers could not be classified. Previous exposure to potentially cross-reacting flaviviruses (Saint Louis Encephalitis [SLE] and Yellow Fever [YF] viruses) was analyzed, as a possible cause of the secondary response pattern. Our results indicated that among cases classified as dengue secondary response, 83

could be attributed to previous SLE or YF exposure or serologic cross-reactivity. Vaccination against YF virus was at most a minor contributor to the secondary response pattern. The finding of a positive YF serologic result among persons not vaccinated may indicate silent circulation of YF in a region that can support both urban and jungle cycles. Other cases showing dengue secondary responses remained unexplained, suggesting the unrecognized occurrence of a previous infection with other dengue serotypes or of flaviviruses other than SLE or YF.