Clonal haematopoiesis - a novel entity that modifies pathological processes in elderly

Abstract Progress in the development of new sequencing techniques with wider accessibility and higher sensitivity of the protocol of deciphering genome particularities led to the discovery of a new phenomenon – clonal haematopoiesis. It is characterized by the presence in the bloodstream of elderly people a minor clonal population of cells with mutations in certain genes, but without any sign of disease related to the hematopoietic system. Here we will review this recent advancement in the field of clonal haematopoiesis and how it may affect the disease’s development in old age

A novel bifunctional hybrid with marine bacterium alkaline phosphatase and Far Eastern holothurian mannan-binding lectin activities.

A fusion between the genes encoding the marine bacterium Cobetia marina alkaline phosphatase (CmAP) and Far Eastern holothurian Apostichopus japonicus mannan-binding C-type lectin (MBL-AJ) was performed. Expression of the fusion gene in E. coli cells resulted in yield of soluble recombinant chimeric protein CmAP/MBL-AJ with the high alkaline phosphatase activity and specificity of the lectin MBL-AJ. The bifunctional hybrid CmAP/MBL-AJ was produced as a dimer with the molecular mass of 200 kDa. The CmAP/MBL-AJ dimer model showed the two-subunit lectin part that is associated with two molecules of alkaline phosphatase functioning independently from each other. The highly active CmAP label genetically linked to MBL-AJ has advantaged the lectin-binding assay in its sensitivity and time. The double substitution A156N/F159K in the lectin domain of CmAP/MBL-AJ has enhanced its lectin activity by 25 ± 5%. The bifunctional hybrid holothurian's lectin could be promising tool for developing non-invasive methods for biological markers assessment, particularly for improving the MBL-AJ-based method for early detection of a malignant condition in cervical specimens

Data supporting functional diversity of the marine bacterium Cobetia amphilecti KMM 296

Data is presented in support of functionality of hyper-diverse protein families encoded by the Cobetia amphilecti KMM 296 (formerly Cobetia marina KMM 296) genome (“The genome of the marine bacterium Cobetia marina KMM 296 isolated from the mussel Crenomytilus grayanus (Dunker, 1853)” [1]) providing its nutritional versatility, adaptability and biocontrol that could be the basis of the marine bacterium evolutionary and application potential. Presented data include the information of growth and biofilm-forming properties of the food-associated isolates of Pseudomonas, Bacillus, Listeria, Salmonella and Staphylococcus under the conditions of their co-culturing with C. amphilecti KMM 296 to confirm its high inter-species communication and anti-microbial activity. Also included are the experiments on the crude petroleum consumption by C. amphilecti KMM 296 as the sole source of carbon in the presence of sulfate or nitrate to ensure its bioremediation capacity. The multifunctional C. amphilecti KMM 296 genome is a promising source for the beneficial psychrophilic enzymes and essential secondary metabolites

Recombinant Fusion Protein Joining E Protein Domain III of Tick-Borne Encephalitis Virus and HSP70 of Yersinia pseudotuberculosis as an Antigen for the TI-Complexes

Domain III (DIII) of the tick-borne encephalitis virus (TBEV) protein E contains epitopes, which induce antibodies capable of neutralizing the virus. To enhance the immunogenicity of this protein, which has a low molecular weight, the aim of the present work was to express, isolate, and characterize a chimeric protein based on the fusion of the bacterial chaperone HSP70 of Yersinia pseudotuberculosis and EIII (DIII + stem) as a prospective antigen for an adjuvanted delivery system, the tubular immunostimulating complex (TI-complex). The chimeric construction was obtained using pET-40b(+) vector by ligating the respective genes. The resulting plasmid was transformed into DE3 cells for the heterologous expression of the chimeric protein, which was purified by immobilized metal affinity chromatography (IMAC). ELISA, differential scanning calorimetry, intrinsic fluorescence, and computational analysis were applied for the characterization of the immunogenicity and conformation of the chimeric protein. Mice immunization showed that the chimeric protein induced twice the number of anti-EIII antibodies in comparison with EIII alone. In turn, the incorporation of the HSP70/EIII chimeric protein in the TI-complex resulted in a twofold increase in its immunogenicity. The formation of this vaccine construction was accompanied by significant conformational changes in the chimeric protein. Using HSP70 in the content of the chimeric protein represents an efficient means for presenting the main antigenic domain of the TBEV envelope protein to the immune system, whereas the incorporation of this chimeric protein into the TI-complex further contributes to the development of a stronger immune response against the TBEV infection

The homology model of <i>Cm</i>AP monomer with binding sites for two Zn²⁺ and one Mg²⁺ ions.

<p>The homology model of <i>Cm</i>AP monomer with binding sites for two Zn²⁺ and one Mg²⁺ ions.</p

The homology model of MBL-<i>AJ</i> monomer (A), the superimposition of MBL-<i>AJ</i> model with template CEL-IV (B) and the structural model of MBL-<i>AJ</i> dimer (C).

<p>(a) - Graphic representation of MBL-<i>AJ</i> monomer with disulfide bonds (Cys24-Cys35, Cys52-Cys168) and free Cys60 and Cys172 group at C-terminus of lectin displayed as sticks. (b) - Superimposition of the MBL-<i>AJ</i> structural model (orange) and template CEL-IV (purple). Loop L of MBL-<i>AJ</i> (orange) with significant difference is indicated. (c) – The structural model of MBL-<i>AJ</i> dimer build from monomer M1 (orange) and M2 (purple) with two intermonomer disulfide bonds Cys′60-Cys172 and Cys60-Cys′172.</p

SDS-PAGE of <i>Cm</i>AP/MBL-<i>AJ</i> eluted from Superdex 200: 1 –molecular weight marker (Bio-rad); 2 –<i>Cm</i>AP/MBL-<i>AJ</i>.

<p>SDS-PAGE of <i>Cm</i>AP/MBL-<i>AJ</i> eluted from Superdex 200: 1 –molecular weight marker (Bio-rad); 2 –<i>Cm</i>AP/MBL-<i>AJ</i>.</p

2D-Diagram of the oligosaccharide molecular contacts with A155N/F159K MBL-AJ mutant and contacts of model oligosaccharide with A155N/F159K MBL-AJ mutant.

<p>2D-Diagram of the oligosaccharide molecular contacts with A155N/F159K MBL-AJ mutant and contacts of model oligosaccharide with A155N/F159K MBL-AJ mutant.</p