Proteomic analysis in cerebrospinal fluid of patients with atypical nonketotic hyperglycinemia and pulmonary hypertension - A pilot study

A variant phenotype of nonketotic hyperglycinemia has been described by our group associated with pulmonary hypertension. The aim of this study is to investigate the cerebrospinal fluid proteomes to get an insight into this neurodegenerative process producing leukoencephalopathy with white matter spongiform degeneration. DIGE and MALDI-TOF-TOF analyses were performed to carry out the proteomic study of four patients against three normal controls and one additional control of a classical nonketotic hyperglycinemia. The differential proteomic analysis showed a displacement of some series of spots toward the acidic side. The shifted proteins showed a high degree of carbonylation and increased methionine sulfoxidation was found in cystatin C and in vitamin-D-binding protein. These findings in addition to the increase of serum malondialdehyde concentration provide evidence of an oxidative stress in the patients under study, which is probably systemic rather than mainly confined to the CNS. The similarities of our findings with those found in other neurodegenerative diseases suggest that oxidative damage is commonly involved in these pathologies. DIGE technology improves the 2-D PAGE differential analysis and it is suitable in proteomic studies with a small number of cases.     

Phase and morphology evolution of calcium carbonate precipitated by carbonation of hydrated lime

Phase and morphology evolution of CaCO₃ precipitated during carbonation of lime pastes via the reaction Ca(OH)₂ + CO₂ → CaCO₃ + H₂O has been investigated under different conditions (pCO₂ ≈ 10^−3.5 atm at 60 % RH and 93 % RH; pCO₂ = 1 atm at 93 % RH) using XRD, FTIR, TGA, and SEM. Simulations of the pore solution chemistry for different stages and conditions of carbonation were performed using the PHREEQC code to investigate the evolution of the chemistry of the system. Results indicate initial precipitation of amorphous calcium carbonate (ACC) which in turn transforms into scalenohedral calcite under excess Ca²⁺ ions. Because of their polar character, { 21[`3]4 } \left\{ {21\bar{3}4} \right\} scalenohedral faces (type S) interact more strongly with excess Ca<sup>2+</sup> than non-polar { 10[`1]4 } \left\{ {10\bar{1}4} \right\} rhombohedral faces (type F), an effect that ultimately favors the stabilization of { 21[`3]4 } \left\{ {21\bar{3}4} \right\} faces. Following the full consumption of Ca<sup>2+</sup> ions and further dissolution of CO<sub>2</sub> leading to a pH drop of the pore solution, { 21[`3]4 } \left\{ {21\bar{3}4} \right\} scalenohedra are subjected to dissolution. This eventually results in re-precipitation of { 10[`1]4 } \left\{ {10\bar{1}4} \right\} rhombohedra at close-to-neutral pH. This crystallization sequence progresses through the carbonated depth with a strong dependence on the degree of exposure to CO₂, which is controlled by the carbonated pore structure governing the diffusion of CO₂. Both the carbonation process and the scalenohedral-to-rhombohedral transformation are kinetically favored under high RH and high pCO₂. Supersaturation plays a critical role on the nucleation density and size of CaCO₃ crystals. These results have important implications in understanding the behavior of ancient and modern lime mortars for applications in architectural heritage conservation.     

Synergistic Effect on Wettability of Mixtures of Amine Oxides,Alkylpolyglucosides, and Ethoxylated Fatty Alcohols

The mixture proportions of three surfactants, i.e. an alkylpolyglucoside (APG), an ethoxylated fatty alcohol, and an amine oxide (AO), were optimized by applying Response Surface Methodology in order to achieve the surfactant ratio that produces the highest wetting power. The synergistic effect of the binary samples of AO with the other surfactants was firstly verified. An improvement in this synergistic effect on wettability was found in the ternary mixture. In the experimental range analyzed, the surfactant concentration ratio that produced the highest wettability was a composition with 19.3 % APG, 30.1 % ethoxylated alcohol, and 50.6 % lauramine oxide.     

Experimental Design for Optimizing the Cleaning of Starch Adhering to Stainless-Steel Surfaces Using Nonionic Surfactants and Silica Microparticles

The statistical design of experiments has been used to assess the detergency of starch adhering to stainless steel considering three factors: pH, flow rate, and concentration of silica particles in the cleaning solution. The cleaning tests were carried out in a continuous-flow device that simulates the behavior of a Cleaning-In-Place washing system. Different statistical designs were used to evaluate the detergency of cleaning solutions in the absence of surfactants and with two nonionic surfactants, i.e., an ethoxylated alcohol, and an alkylpolyglucoside (APG). Expressions were developed to simulate detergency levels as a function of the variables assayed, determining the optimal detergency of each cleaning solution studied. The results indicate that the variable most influential on detergency was pH. Cleaning solutions with high alkalinity are required to achieve a significant cleaning efficiency. On the other hand, the silica-particle concentration did not influence the detergency results using cleaning solutions without surfactants. Nevertheless, the addition of either the ethoxylated alcohol or the APG to the cleaning solution with silica particles has a significant effect on detergency: it diminished with the ethoxylated alcohol and increased with the APG. Maximum detergency was found at the highest level of pH, particle concentration, and flow rate when the APG was added to the cleaning solution.     

Chromosome instability and unusual features of some widely used strains of Candida albicans

Electrophoretic karyotyping of the Candida albicans revealed a different migration pattern of ChR in three different stocks of the sequencing strain SC5314. In one stock, the high instability of ChR size prevented the migration of ChR as a compact band; ChR appeared, instead, as a smear. In some stocks, ChR and/or Ch1 ploidy diminished, suggesting mixed populations of disomic and monosomic cells. Similarly, some stocks of widely used derivatives CAI4 and BWP17 contained smearing of ChR. In addition, the most manipulated strain in the lineage of SC5314, the last derivative, BWP17, acquired an increase in the size of Ch7b and revealed an unusual property. BWP17 did not tolerate a well-established procedure of telomere-mediated fragmentation of a chromosome; the remaining intact homologue always duplicated. We suggest that some stocks of SC5314 are unstable and that BWP17 may not be appropriate for general studies. Instead of BWP17 or CAI4, we recommend using for general research CAF4-2, which is a relatively stable Ura- derivative, and which has been successfully used for more than a decade in our laboratory.     

Assessing methods for blood cell cytotoxic responses to inorganic nanoparticles and nanoparticle aggregates

Inorganic nanoparticles (NPs) show great potential for medicinal therapy. However, biocompatibility studies are essential to determine if they are safe. Here, five different NPs are compared for their cytotoxicity, internalization, aggregation in medium, and reactive oxygen species (ROS) production, using tumoral and normal human blood cells. Differences depending on the cell type are analyzed, and no direct correlation between ROS production and cell toxicity is found. Results are discussed with the aim of standardizing the procedures for the evaluation of the toxicity.