Enzymatic Hydrolysis of Sunflower Oil: Characterisation of Interface

The results of a study of the enzymatic hydrolysis of sunflower oil are presented. The influence of the enzyme and substrate concentrations has been investigated. The interfacial area was modified by changing the volume fraction of oil, while keeping other variables constant. A phase inversion is indicated, which influenced the reaction kinetics by the possible formation of enzyme multilayers, because of steric hindrance. By means of the initial hydrolysis rate, the lipase concentration for achieving interface saturation was estimated. A Lewis cell type contactor was used for the determination of the interface surface, and droplet diameters were deduced. © 1997 SCI.     

BaGdF5 Nanophosphors Doped with Different Concentrations of Eu3+ for Application in X-ray Photodynamic Therapy

X-ray photodynamic therapy (XPDT) has been recently considered as an efficient alternative to conventional radiotherapy of malignant tissues. Nanocomposites for XPDT typically consist of two components—a nanophosphor which re-emits X-rays into visible light that in turn is absorbed by the second component, a photosensitizer, for further generation of reactive oxygen species. In this study, BaGdF₅ nanophosphors doped with different Eu:Gd ratios in the range from 0.01 to 0.50 were synthesized by the microwave route. According to transmission electron microscopy (TEM), the average size of nanophosphors was ~12 nm. Furthermore, different coatings with amorphous SiO₂ and citrates were systematically studied. Micro-CT imaging demonstrated superior X-ray attenuation and sufficient contrast in the liver and the spleen after intravenous injection of citric acid-coated nanoparticles. In case of the SiO₂ surface, post-treatment core–shell morphology was verified via TEM and the possibility of tunable shell size was reported. Nitrogen adsorption/desorption analysis revealed mesoporous SiO₂ formation characterized by the slit-shaped type of pores that should be accessible for methylene blue photosensitizer molecules. It was shown that SiO₂ coating subsequently facilitates methylene blue conjugation and results in the formation of the BaGdF₅: 10% Eu³⁺@SiO₂@MB nanocomposite as a promising candidate for application in XPDT.     

Acid Sphingomyelinase Deficiency: A Clinical and Immunological Perspective

Acid sphingomyelinase deficiency (ASMD) is a lysosomal storage disease caused by deficient activity of acid sphingomyelinase (ASM) enzyme, leading to the accumulation of varying degrees of sphingomyelin. Lipid storage leads to foam cell infiltration in tissues, and clinical features including hepatosplenomegaly, pulmonary insufficiency and in some cases central nervous system involvement. ASM enzyme replacement therapy is currently in clinical trial being the first treatment addressing the underlying pathology of the disease. Therefore, presently, it is critical to better comprehend ASMD to improve its diagnose and monitoring. Lung disease, including recurrent pulmonary infections, are common in ASMD patients. Along with lung disease, several immune system alterations have been described both in patients and in ASMD animal models, thus highlighting the role of ASM enzyme in the immune system. In this review, we summarized the pivotal roles of ASM in several immune system cells namely on macrophages, Natural Killer (NK) cells, NKT cells, B cells and T cells. In addition, an overview of diagnose, monitoring and treatment of ASMD is provided highlighting the new enzyme replacement therapy available.     

The Role of Pectobacterium atrosepticum Exopolysaccharides in Plant–Pathogen Interactions

The phytopathogenic bacterium Pectobacterium atrosepticum (Pba), one of the members of the soft rot Pectobacteriaceae, forms biofilm-like structures known as bacterial emboli when colonizing the primary xylem vessels of the host plants. The initial extracellular matrix of the bacterial emboli is composed of the host plant’s pectic polysaccharides, which are gradually substituted by the Pba-produced exopolysaccharides (Pba EPS) as the bacterial emboli “mature”. No information about the properties of Pba EPS and their possible roles in Pba-plant interactions has so far been obtained. We have shown that Pba EPS possess physical properties that can promote the maintenance of the structural integrity of bacterial emboli. These polymers increase the viscosity of liquids and form large supramolecular aggregates. The formation of Pba EPS aggregates is provided (at least partly) by the acetyl groups of the Pba EPS molecules. Besides, Pba EPS scavenge reactive oxygen species (ROS), the accumulation of which is known to be associated with the formation of bacterial emboli. In addition, Pba EPS act as suppressors of the quantitative immunity of plants, repressing PAMP-induced reactions; this property is partly lost in the deacetylated form of Pba EPS. Overall, our study shows that Pba EPS play structural, protective, and immunosuppressive roles during Pba–plant interactions and thus should be considered as virulence factors of these bacteria.     

Sensitive Immunofluorescent Detection of the PRAME Antigen Using a Practical Antibody Conjugation Approach

Bioconjugation of antibodies with various payloads has diverse applications across various fields, including drug delivery and targeted imaging techniques. Fluorescent immunoconjugates provide a promising tool for cancer diagnostics due to their high brightness, specificity, stability and target affinity. Fluorescent antibodies are widely used in flow cytometry for fast and sensitive identification and collection of cells expressing the target surface antigen. Nonetheless, current approaches to fluorescent labeling of antibodies most often use random modification, along with a few rather sophisticated site-specific techniques. The aim of our work was to develop a procedure for fluorescent labeling of immunoglobulin G via periodate oxidation of antibody glycans, followed by oxime ligation with fluorescent oxyamines. Here, we report a novel technique based on an in situ oxime ligation of ethoxyethylidene-protected aminooxy compounds with oxidized antibody glycans. The approach is suitable for easy modification of any immunoglobulin G, while ensuring that antigen-binding domains remain intact, thus revealing various possibilities for fluorescent probe design. The technique was used to label an antibody to PRAME, a cancer-testis protein overexpressed in a number of cancers. A 6H8 monoclonal antibody to the PRAME protein was directly modified with protected-oxyamine derivatives of fluorescein-type dyes (FAM, Alexa488, BDP-FL); the stoichiometry of the resulting conjugates was characterized spectroscopically. The immunofluorescent conjugates obtained were applied to the analysis of bone marrow samples from patients with oncohematological diseases and demonstrated high efficiency in flow cytometry quantification. The approach can be applied for the development of various immunofluorescent probes for detection of diagnostic and prognostic markers, which can be useful in anticancer therapy.     

New Insights into the Structure of Kappa/Beta-Carrageenan: A Novel Potential Inhibitor of HIV-1

New insights into the structure of the hybrid κ/β-carrageenan (κ/β-CRG) of the red alga Tichocarpus crinitus have been obtained. Carrageenan oligosaccharides were prepared through the chemical and enzymatic depolymerization of κ/β-CRG with κ-carrageenase and its the enzyme-resistant fraction. The composition and distribution of the repetition units of κ/β- CRG were investigated by using the negative ion tandem MALDI-TOFMS and ESIMS method, which made it possible to prove and characterize the hybrid structure of this polysaccharide. An analysis revealed the blockwise distribution of the long β-blocks along the polysaccharide chain, with the inclusion of κ/β, μ/ν-blocks and some ι-blocks. Furthermore, the desulfated κ/β-CRG was shown to contain of –G–D– repeating units up to 3.5 kDa. Previous studies have demonstrated that CRGs suppress the replication of several viruses. Here, we established that κ/β-CRG and its oligosaccharides significantly inhibit the transduction efficiency of replication-defective lentiviral particles pseudotyped with the envelope proteins of three different viruses. We found that the polysaccharide and its oligosaccharides strongly reduced the transduction efficiency of lentiviral particles pseudotyped with GP160—the envelope protein of the human immunodeficiency virus HIV-1—when added to T-lymphocyte Jurkat cells. The CRG oligosaccharides displayed significantly higher antiviral activity.     

Visualization and quantification of forced in-plane flow through deformed porous media

Fluid flow in liquid molding processes and in other applications involving porous media is often characterized with a permeability tensor and modeled by d'Arcy's law. The permeability is a sensitive function of pore structure, which, in deformable materials, is influenced by compression and extension. The majority of previous permeability measurements on composite reinforcement fabrics did not account for deformations imposed on them by corners and curves in the mold. In the present study, transparent molds were designed with a single 90° bend in which the gap between plates was held constant throughout the flow path. Thus, the effects of fabric curvature on permeability were investigated independently of the effects of fabric compression in the thickness direction. A new experimental system was developed to visualize and quantify fluid flow in fabrics mounted in transparent molds. The reported measurements were conducted with fluid flowing through a (vertical) flat region of fabric, around the 90° curve, and then along a second (horizontal) flat region. Permeability was found to be reduced by the imposed curvature for a nonwoven polyester and three-dimensional woven glass fabric. We were able to quantify the effect of curvature on permeability for the former, but not for the latter because of an enduring, dominant nonlinear entrance resistance. For the glass, comparison of two flow rates indicates that the faster flow is characterized by a two stage filling process, whereas, at the slower rate, the liquid front fills all pores at the same time.     

Synthesis and Calculation of Properties of N-Difluoroaminoazoles,the Novel Type of Energetic Materials

A novel class of energetic materials, N-difluoroaminonitroazoles was synthesized. One molecule of these compounds contains simultaneously a nitro group and difluoroamine one. A method of synthesizing N-difluoroaminonitroazoles incorporates the interaction of N-unsubstituted nitroazoles with O-fluorosulfonyl-N,N-difluorohydroxylamine (F₂NOSO₂F) in the presence of bases. To estimate possible prospects of these compounds, energy content, molecular crystal density, some parameters of detonation and combustion were calculated using some well-known methods of computational chemistry, as well as some of our original schemes. In addition, thermostability (by the DTA method) and density were identified experimentally for a cross-section row of the studied compounds. The reasons for relatively low stability of this type of substances were given.     

Prolonging Shelf-Life of Carp by Combined Ionising Radiation and Refrigeration

Experiments were performed using ionisating radiation in order to prolong the shelf-life of two groups of refrigerated fish ( Cyprinus carpio ). After irradiation fish were kept at 0–2°C. Non-irradiated fish reached the non-acceptability point in 16 days and irradiated fish reached that point in 31 days. No difference was found between shelf-life of whole or eviscerated fish. If fish were immediately cooled to 0°C after death, their shelf-life was prolonged considerably. From this study it is clear that chemical tests of freshness such as TVB-N and K value determination are not appropriate for the study of irradiated fish. Organoleptic estimations and a new determination using an odour concentration meter seem to fit best the objective determination of freshness.