Computer-based engineering of thermostabilized antibody fragments

We used the molecular modeling program Rosetta to identify clusters of amino acid substitutions in antibody fragments (scFvs and scAbs) that improve global protein stability and resistance to thermal deactivation. Using this methodology, we increased the melting temperature (T_m) and resistance to heat treatment of an antibody fragment that binds to the Clostridium botulinum hemagglutinin protein (anti-HA33). Two designed antibody fragment variants with two amino acid replacement clusters, designed to stabilize local regions, were shown to have both higher T_m compared to the parental scFv and importantly to retain full antigen binding activity after 2 hr of incubation at 70°C. The crystal structure of one thermostabilized scFv variants was solved at 1.6 Å and shown to be in close agreement with the RosettaAntibody model prediction.     

Antifungal Activity of Lactic Acid Bacteria Isolated from Peanuts,Gari, and Orange Fruit Juice against Food Aflatoxigenic Molds

This study aims to evaluate the antifungal activity of lactic acid bacteria (LAB) from some Cameroonian food commodities against mycotoxigenic and spoilage molds. Following LAB isolation, the antifungal activity of the isolates was assessed. The organic acids were quantified using high-performance liquid chromatography and the ability of the LAB to reduce mold biomass and aflatoxin production was evaluated. The LAB were identified and the biopreservative potential of strain LO3 was evaluated on tomato paste. Nine percent of the strains isolated showed broad antifungal activity. The activity was due to the effect of organic acids comprising lactic, acetic, 4-hydroxy-3-phenyllactic and 3-phenyllactic acids. Lactobacillus plantarum LO3 exhibiting the highest and broadest antifungal activity was selected and showed the capacity to inhibit fungal growth and aflatoxin production in vitro. Moreover, this strain and its cell-free supernatant showed the ability to prevent aflatoxigenic mold growth in tomato paste without altering its physico-chemical and organoleptic properties.     

Foam-like scaffolds for bone tissue engineering based on a novel couple of silicate-phosphate specular glasses: synthesis and properties

Glass–ceramic scaffolds mimicking the structure of cancellous bone were produced via sponge replication technique by using a polyurethane foam as template and glass powder below 30 μm as inorganic phase. Specifically, a SiO₂-based glass of complex composition and its corresponding P₂O₅-based “specular” glass were used as materials for scaffolding. The polymeric sponge was thermally removed and the glass powders were sintered to obtain a replica of the template structure. The scaffolds were investigated and compared from a structural, morphological and mechanical viewpoint by assessing their crystalline phases, volumetric shrinkage, pores content and interconnection, mechanical strength. In addition, the scaffolds were soaked in acellular simulated body fluid to investigate their in vitro behaviour. The produced scaffolds have a great potential for bone reconstructive surgery because their features, such as shape, strength, bioactivity and bioresorption, can be easily tailored according to the end use.     

Characterisation of optically driven microstructures for manipulating single DNA molecules under a fluorescence microscope

Optical tweezers are powerful tools for manipulating single DNA molecules using fluorescence microscopy, particularly in nanotechnology‐based DNA analysis. We previously proposed a manipulation technique using microstructures driven by optical tweezers that allows the handling of single giant DNA molecules of millimetre length that cannot be manipulated by conventional techniques. To further develop this technique, the authors characterised the microstructures quantitatively from the view point of fabrication and efficiency of DNA manipulation under a fluorescence microscope. The success rate and precision of the fabrications were evaluated. The results indicate that the microstructures are obtained in an aqueous solution with a precision ∼50 nm at concentrations in the order of 10⁶ particles/ml. The visibility of these microstructures under a fluorescence microscope was also characterised, along with the elucidation of the fabrication parameters needed to fine tune visibility. Manipulating yeast chromosomal DNA molecules with the microstructures illustrated the relationship between the efficiency of manipulation and the geometrical shape of the microstructure. This report provides the guidelines for designing microstructures used in single DNA molecule analysis based on on‐site DNA manipulation, and is expected to broaden the applications of this technique in the future.Inspec keywords: DNA, molecular biophysics, fluorescence, optical microscopy, radiation pressure, biological techniquesOther keywords: optically driven microstructures, single DNA molecule analysis, fluorescence microscopy, optical tweezers, nanotechnology‐based DNA analysis, manipulation technique, aqueous solution, fine tune visibility, yeast chromosomal DNA molecules, geometrical shape, on‐site DNA manipulation     

Cancer Stem Cells: Biological Functions and Therapeutically Targeting

Almost all tumors are composed of a heterogeneous cell population, making them difficult to treat. A small cancer stem cell population with a low proliferation rate and a high tumorigenic potential is thought to be responsible for cancer development, metastasis and resistance to therapy. Stem cells were reported to be involved in both normal development and carcinogenesis, some molecular mechanisms being common in both processes. No less controversial, stem cells are considered to be important in treatment of malignant diseases both as targets and drug carriers. The efforts to understand the role of different signalling in cancer stem cells requires in depth knowledge about the mechanisms that control their self-renewal, differentiation and malignant potential. The aim of this paper is to discuss insights into cancer stem cells historical background and to provide a brief review of the new therapeutic strategies for targeting cancer stem cells.     

Interfacial polymerization of an epoxy resin and carboxylic acids for the synthesis of microcapsules

BACKGROUND: Microencapsulation technology promises new applications such as intelligent microstructures, phase change materials and self‐healing composites. Microcapsule synthesis and characterization have been researched extensively; however, the well‐known polymerization between epoxy resins and carboxylic acids has not been used to prepare microcapsules. RESULTS: Microcapsules were prepared by interfacial polymerization of an oil‐in‐water emulsion which contained a commercial epoxy resin and multifunctional carboxylic acids. The microcapsules obtained were characterized using optical microscopy and scanning electron microscopy. Experiments performed at lower stirring rates led to larger microcapsules, in the range 100–400 µm, while higher stirring rates resulted in microcapsules in the range 10–50 µm. CONCLUSIONS: Microcapsules can be prepared by interfacial polymerization of epoxy resins, an extensively studied and widely used class of polymers. By means of NMR characterization we gained insight into the mechanism of polymerization at the interface wherein products coming from the more hindered ring opening as well as from intermolecular transesterification are identified. The presence of a crosslinker affects the morphology of the external microcapsule surface. Copyright © 2008 Society of Chemical Industry     

Action of some organophosphonic compounds on a diglycidyl ether-bisphenol-A epoxy resin

The effect of three organophosphonic compounds on the thermooxidative degradation of the uncured diglycidyl ether-bisphenol-A epoxy resin was investigated, using infrared spectroscopy, thermogravimetry, differential thermogravimetry, static pyrolysis, pyrolysis-gas chromatography and elemental analysis. The results obtained indicate that additives with phosphorus and chlorine or nitrogen in molecules exercise an influence upon the degradative process of uncured resin by catalysing its decomposition in the lower temperature region. At the same time, the tested additives promote the formation of highly thermostable and non-flammable structures in the solid residue.