Thermal Stability Threshold for Amyloid Formation in Light Chain Amyloidosis

Light chain (AL) amyloidosis is a devastating disease characterized by amyloid deposits formed by immunoglobulin light chains. Current available treatments involve conventional chemotherapy and autologous stem cell transplant. We have recently concluded a phase III trial comparing these two treatments. AL amyloidosis patients who achieve hematological complete response (CR) do not necessarily achieve organ response regardless of the treatment they received. In order to investigate the possible correlation between amyloid formation kinetics and organ response, we selected AL amyloidosis patients from the trial with kidney involvement and CR after treatment. Six patients were selected and their monoclonal immunoglobulin light chains were characterized. The proteins showed differences in their stability and their kinetics of amyloid formation. A correlation was detected at pH 7.4, showing that less stable proteins are more likely to form amyloid fibrils. AL-T03 is too unstable to form amyloid fibrils at pH 7.4. This protein was found in the only patient in the study that had organ response, suggesting that partially folded species are required for amyloid formation to occur in AL amyloidosis.     

β-Cyclodextrin Inclusion Complex to Improve Physicochemical Properties of Pipemidic Acid: Characterization and Bioactivity Evaluation

The aptitude of cyclodextrins (CDs) to form host-guest complexes has prompted an increase in the development of new drug formulations. In this study, the inclusion complexes of pipemidic acid (HPPA), a therapeutic agent for urinary tract infections, with native β-CD were prepared in solid state by kneading method and confirmed by FT-IR and ¹H NMR. The inclusion complex formation was also characterized in aqueous solution at different pH via UV-Vis titration and phase solubility studies obtaining the stability constant. The 1:1 stoichiometry was established by a Job plot and the inclusion mechanism was clarified using docking experiments. Finally, the antibacterial activity of HPPA and its inclusion complex was tested on P. aeruginosa, E. coli and S. aureus to determine the respective EC_50s and EC_90s. The results showed that the antibacterial activity of HPPA:β-CD against E. coli and S. aureus is higher than that of HPPA. Furthermore, HPPA and HPPA:β-CD, tested on human hepatoblastoma HepG2 and MCF-7 cell lines by MTT assay, exhibited, for the first time, antitumor activities, and the complex revealed a higher activity than that of HPPA. The use of β-CD allows an increase in the aqueous solubility of the drug, its bioavailability and then its bioactivity.     

Multi‐phosphorylation of the Intrinsically Disordered Unique Domain of c‐Src Studied by In‐Cell and Real‐Time NMR Spectroscopy

Intrinsically disordered regions (IDRs) are preferred sites for post‐translational modifications essential for regulating protein function. The enhanced local mobility of IDRs facilitates their observation by NMR spectroscopy in vivo. Phosphorylation events can occur at multiple sites and respond dynamically to changes in kinase–phosphatase networks. Here we used real‐time NMR spectroscopy to study the effect of kinases and phosphatases present in Xenopus oocytes and egg extracts on the phosphorylation state of the “unique domain” of c‐Src. We followed the phosphorylation of S17 in oocytes, and of S17, S69, and S75 in egg extracts by NMR spectroscopy, MS, and western blotting. Addition of specific kinase inhibitors showed that S75 and S69 are phosphorylated by CDKs (cyclin‐dependent kinases) differently from Cdk1. Moreover, although PKA (cAMP‐dependent protein kinase) can phosphorylate S17 in vitro, this was not the major S17 kinase in egg extracts. Changes in PKA activity affected the phosphorylation levels of CDK‐dependent sites, thus suggesting indirect effects of kinase–phosphatase networks. This study provides a proof‐of‐concept of the use of real‐time in vivo NMR spectroscopy to characterize kinase/phosphatase effects on intrinsically disordered regulatory domains.     

Controlled release matrices and micro/nanoparticles of chitosan with antimicrobial potential: development of new strategies for microbial control in agriculture

The control of micro‐organisms responsible for pre‐ and postharvest diseases of agricultural products, mainly viruses and fungi, is a problem that remains unresolved, together with the environmental impact of the excessive use of chemicals to tackle this problem. Current efforts are focused on the search for efficient alternatives for microbial control that will not result in damage to the environment or an imbalance in the existing biota. One alternative is the use of natural antimicrobial compounds such as chitosan, a linear cationic biopolymer, which is biodegradable, biocompatible and non‐toxic, has filmogenic properties and is capable of forming matrices for the transport of active substances. The study of chitosan has attracted great interest owing to its ability to form complexes or matrices for the controlled release of active compounds such as micro‐ and nanoparticles, which, together with the biological properties of chitosan, has allowed a major breakthrough in the pharmaceutical and biomedical industries. Another important field of study is the development of chitosan‐based matrices for the controlled release of active compounds in areas such as agriculture and food for the control of viruses, bacteria and fungi, which is one of the least exploited areas and holds much promise for future research. © 2013 Society of Chemical Industry     

Macroporous calcium phosphate ceramic implants for sustained drug delivery

Limited blood circulation to the skeletal tissue is a major cause of reduced therapeutic effects seen with drugs administered by conventional systemic ways. To resolve this issue and obtain a sufficiently high local concentration to induce therapeutic effects, several implanted drug delivery systems have been developed for hard tissues using biomaterials.We have designed a drug delivery device based on hydroxylapatite (HA) and tested it in vitro using metronidazol and chloramphenicol as model compounds. Porous HA ceramics were prepared with two different shapes (cylindrical and spherical). Known amounts of drugs were introduced inside a drilled hole and sealed with wax. The ceramics were then suspended in stirred distilled water in closed polypropylene vials. Drug release was observed during several weeks.A mathematical model used to describe drug release from HA was elaborated based on the expressions of Fick's laws. The experimental kinetic results could be related to ceramic constitution and to drug localization.     

Enhanced Mechanical Properties and Corrosion Behavior of Biodegradable Mg-Zn/HA Composite

Metallurgical and Materials Transactions A - Magnesium (Mg) and its alloys have shown potential for use in the biomedical industry due to their excellent biological performance and biodegradability...     

Effect of the curing régime on compressive strength

In a previous issue the Instituto Eduardo Torroja in Spain described the development of lightweight panels made from ordinary Portland cement and rice husk ash (RHA). This paper discusses the crucial effect of the curing régime on compressive strength with temperatures maintained at 30C and relative humidity at 80 percent.     

Antioxidant Effects of Berry Phenolics Incorporated in Oil-in-Water Emulsions with Continuous Phase β-Lactoglobulin

The purpose of this study was to evaluate the effects of berry phenolics, in this case, black currant (Ribes nigrum) anthocyanins and raspberry (Rubus idaeus) ellagitannins, in the presence of continuous phase β-lactoglobulin (β-Lg), on the oxidative stability of Brij 35-stabilized corn oil-in-water emulsions. The extent of lipid oxidation in emulsions was measured by determining the formation of lipid hydroperoxides and hexanal, and extent of protein oxidation by monitoring the loss of tryptophan and cysteine residues in the continuous phase β-Lg. Berry phenolics at concentration levels of 20 and 50 μM were able to prevent lipid oxidation with and without β-Lg in the aqueous phase. The results show that a combination of β-Lg and berry phenolics was more efficient in inhibiting hexanal formation than either component alone. Synergistic effects on antioxidant activity toward hexanal were observed only at the 20 μM levels of berry phenolics in combination with continuous phase β-Lg. The berry phenolics were also able to inhibit the oxidation of tryptophan and cysteine residues of β-Lg. The results show that the amino acid residues were oxidized prior to the propagation of lipid oxidation. This suggests that these amino acids were able to inhibit fatty acid scission. The information gained from this study would be useful in protecting emulsion-based food products from oxidative deterioration.