Detection of the gangliosideN-glycolyl-neuraminyl-lactosyl-ceramide by biotinylatedEscherichia coli K99 lectin

K99 lectin fromEscherichia coli was purified and biotinylated via its carboxyl groups using biocytin hydrazide and a water soluble carbodiimide. Biotinylation of two out of the nine carboxyl groups was sufficient to permit detection of the lectin by avidin and did not cause any loss of the haemagglutinating activity. It was demonstrated that the biotinylated K99 lectin retained other important properties of native K99 and that it will probably become a very sensitive detecting reagent. Indeed, it was able to bind to HeLa cells, as do intact bacteria carrying K99 fimbriae, and also to recognizeN-glycolyl-neuraminyl-lactosyl-ceramide in an overlay binding assay. Abbreviations: NeuAc,N-acetylneuraminic acid; NeuGc,N-glycolylneuraminic acid; PBS, phosphate buffered saline (0.9% NaCl containing 150mm sodium phosphate, pH 7.2); LPS, lipopolysaccharide; BCHZ, biocytin hydrazide; EDC, 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide; BSA, bovine serum albumin; SDS-PAGE, sodium dodecyl sulfate-polyacrylamide gel electrophoresis; DMEM, Dulbecco's modified Eagle medium. For the gangliosides, trivial names and structures are given according to the recommendations in [43]. NeuAc2-3Gal1-4Glc1-1Cer (NeuAc-GM₃); NeuGc2-3Gal1-4Glc1-1Cer (NeuGc-GM₃); GalNAc1-4(NeuAc2-3)Gal1-4Glc1-1Cer (GM₂); NeuAc2-8NeuAc2-3Gal1-4Glc1-1Cer (GD3); Gal1-3GalNAc1-4(NeuAc2-3)Gal1-4Glc1-1Cer (GM₁); NeuAc2-3Gal1-3GalNAc1-4(NeuAc2-3)Gal1-4Glc1-1Cer (GD_1a); Gal1-3GalNAc1-4(NeuAc2-8NeuAc2-3)Gal1-4Gle1-1Cer (GD_1b); NeuAc2-3Gal1-3GalNAc1-4(NeuAc2-8NeuAc2-3) Gal1.-4Glc1-1Cer (GT_1b). NeuGc2-3Gal1-4GleNAc1-4Gal1-4Glc1-1Cer (NeuGc-SPG).     

Theoretical study of Escherichia coli peptide deformylase inhibition by several drugs

Because peptide deformylase (PDF) is essential for the initiation of translation in eubacteria but not in eukaryotes, it is a potentially interesting target for antibiotics. Computer simulation using docking software can be used to model protein-ligand interactions, and in this brief report we describe its use in optimizing the design in PDF-directed inhibitors. PDF was used as target for a set of five inhibitors with substantial structural differences. Docking results show that the compound 1BB2 (actinonin) binds with high affinity to the enzyme and produces the most stable complex, forming nine hydrogen bonds with the enzyme active site. Its binding energy is DeltaG = -31.880 kJ/mol. The modeling study shows that when the methyl group of 1BB2 is replaced with an amine group, the binding energy is increased to -35.316 kJ/mole. This enhancement is more marked (DeltaG = -41.141 kJ/mol) when the propyl group and the five-membered ring of 1BB2 are replaced by an amide group and a phenyl ring, respectively. We describe an attempt to design better antibiotics on the basis of a computer-aided simulation of the interaction between a drug and its target molecule.     

Constitutive overexpression of immunoidentical forms of PCP-induced Euglena gracilis CYP-450

Environmental pollutants are classically associated with increased drug metabolism. In this report, antibodies that are able to detect mammalian CYP proteins, namely the CYP1A1, CYP1A2, CYP2B1/B2, and CYP3A4 proteins, were used to investigate the expression of CYP-related proteins in Euglena gracilis (EG) cells under normal and PCP-treated conditions and in a EG-cell line adapted to PCP. Compared to normal conditions, the presence of PCP in the culture medium induced elevated levels of EG CYP-like proteins. With the exception of CYP3A4, this overexpression was correlated with expression of additional forms of CYP proteins having, respectively, the same molecular weight but slightly different pIs. Even in EG cells which had lost their PCP-adapted property after having been cultured without PCP, these additional forms were continuously expressed. This observation raised the question about the definition of a biomarker of pollution.     

Trace elements removal ability and antioxidant activity of Phragmites australis (from Algeria)

Abstract

In this study, roots, stems and leaves of the worldwide distributed macrophyte Phragmites australis (common reed) were tested as potential removal and biomonitors of trace elements contamination in sediment. In particular, the concentrations (100, 200, and 500?mg/kg) of the following elements were analyzed: Zn, Cu, Pb, and Fe. Results showed that the amount of concentrations in plant tissues is significantly (p?≤?0.01) dependent on the kind of organ and element. Trace element concentrations decreased according to the pattern of Fe^a (Roots^a > Stems^b > Leaves^b) > Zn^b (Root^a > Leaves^b > Stems^c) > Cu^c (Roots^a > Leaves^b > Stems^c) > Pb^c (Roots^a > Stems^b > Leaves^c), as well as the roots acted as the main centers of bioaccumulation for all elements studied, and stems as the transit organs for translocation from roots to leaves. The major mechanisms employed by the plant were probably phytostabilization on the basis of the calculated Biological Concentration Factor (BCF – metal concentration ratio of plant root to soil); and Translocation Factor (TF – metal concentration ratio of plants roots to above ground part). Finally, due to the low scavenger effect of the radical DPPH, we excluded the hypothesis of the use of antioxidant mechanism in the tolerance of metals.     

CXCR4 inhibitors selectively eliminate CXCR4-expressing human acute myeloid leukemia cells in NOG mouse model

The chemokine receptor CXCR4 favors the interaction of acute myeloid leukemia (AML) cells with their niche but the extent to which it participates in pathogenesis is unclear. Here, we show that CXCR4 expression at the surface of leukemic cells allowed distinguishing CXCR4^high from CXCR4^neg/low AML patients. When high levels of CXCR4 are expressed at the surface of AML cells, blocking the receptor function with small molecule inhibitors could promote leukemic cell death and reduce NOD/Shi-scid/IL-2Rγ^null (NOG) leukemia-initiating cells (LICs). Conversely, these drugs had no efficacy when AML cells do not express CXCR4 or when they do not respond to chemokine CXC motif ligand 12 (CXCL12). Functional analysis showed a greater mobilization of leukemic cells and LICs in response to drugs, suggesting that they target the interaction between leukemic cells and their supportive bone marrow microenvironment. In addition, increased apoptosis of leukemic cells in vitro and in vivo was observed. CXCR4 expression level on AML blast cells and their migratory response to CXCL12 are therefore predictive of the response to the inhibitors and could be used as biomarkers to select patients that could potentially benefit from the drugs.     

SHARE-ABE: an efficient and secure data sharing framework based on ciphertext-policy attribute-based encryption and Fog computing

Attribute-based encryption (ABE) is an access control mechanism that ensures efficient data sharing among dynamic groups of users by setting up access structures indicating who can access what. However, ABE suffers from expensive computation and privacy issues in resource-constrained environments such as IoT devices. In this paper, we present SHARE-ABE, a novel collaborative approach for preserving privacy that is built on top of Ciphertext-Policy Attribute-Based Encryption (CP-ABE). Our approach uses Fog computing to outsource the most laborious decryption operations to Fog nodes. The latter collaborate to partially decrypt the data using an original and efficient chained architecture. Additionally, our approach preserves the privacy of the access policy by introducing false attributes. Furthermore, we introduce a new construction of a collaboration attribute that allows users within the same group to combine their attributes while satisfying the access policy. Experiments and analyses of the security properties demonstrate that the proposed scheme is secure and efficient especially for resource-constrained IoT devices.