Inhibitory Monoclonal Antibodies and Their Recombinant Derivatives Targeting Surface-Exposed Carbonic Anhydrase XII on Cancer Cells

Monoclonal and recombinant antibodies are widely used for the diagnostics and therapy of cancer. They are generated to interact with cell surface proteins which are usually involved in the development and progression of cancer. Carbonic anhydrase XII (CA XII) contributes to the survival of tumors under hypoxic conditions thus is considered a candidate target for antibody-based therapy. In this study, we have generated a novel collection of monoclonal antibodies (MAbs) against the recombinant extracellular domain of CA XII produced in HEK-293 cells. Eighteen out of 24 MAbs were reactive with cellular CA XII on the surface of live kidney and lung cancer cells as determined by flow cytometry. One MAb 14D6 also inhibited the enzymatic activity of recombinant CA XII as measured by the stopped-flow assay. MAb 14D6 showed the migrastatic effect on human lung carcinoma A549 and renal carcinoma A498 cell lines in a ‘wound healing’ assay. It did not reduce the growth of multicellular lung and renal cancer spheroids but reduced the cell viability by the ATP Bioluminescence assay. Epitope mapping revealed the surface-exposed amino acid sequence (35-FGPDGENS-42) close to the catalytic center of CA XII recognized by the MAb 14D6. The variable regions of the heavy and light chains of MAb 14D6 were sequenced and their complementarity-determining regions were defined. The obtained variable sequences were used to generate recombinant antibodies in two formats: single-chain fragment variable (scFv) expressed in E. coli and scFv fused to human IgG1 Fc fragment (scFv-Fc) expressed in Chinese Hamster Ovary (CHO) cells. Both recombinant antibodies maintained the same specificity for CA XII as the parental MAb 14D6. The novel antibodies may represent promising tools for CA XII-related cancer research and immunotherapy.     

Biodegradability of biodiesel fuel of animal and vegetable origin

One of the positive features of biofuel concerning environmental protection is its high biodegradability. Fuel is considered to be biodegradable if not less than 90% of it degrades within 21 days. The aim of this work was to determine the biodegradability of various kinds of fatty acid methyl esters and their mixtures with fossil diesel fuel in natural environments. It was determined that fatty acid methyl esters meet the requirements for biodegradability set by the EU. Of rapeseed oil fatty acid methyl esters (RME), 91.2% degraded within 21 days, compared to 94.2% of rapeseed oil fatty acid ethyl esters, 98.3% of linseed oil fatty acid methyl esters (LSME), 90% of tallow fatty acid methyl esters, and 92.5% of pork lard fatty acid methyl esters (LME), while the amount of degraded fossil diesel fuel reached only 57.3%. The biodegradability of multi‐component biofuels containing RME, LSME and LME is similar; the best is of a mixture of 70% RME, 6% LSME and 24% LME. It was determined that more than 90% of multi‐component biofuel and fossil diesel fuel mixtures degrade within 21 days when they contain 35% and more of multi‐component biofuel.     

A Petri-Net-based modelling approach to railway bridge asset management

Management of a large portfolio of infrastructure assets is a complex and demanding task for transport agencies. Although extensive research has been conducted on probabilistic models for asset management, in particular bridges, focus has been almost exclusively on deterioration modelling. The model being presented in this study tries to reunite a disjointed system by combining deterioration, inspection and maintenance models. A Petri-Net modelling approach is employed and the resulting model consists of a number of different modules each with its own source of data, calibration methodology and functionality. The modules interconnect providing a robust framework. The interaction between the modules can be used to provide meaningful outputs useful to railway bridge portfolio managers.     

Bioconversion of agro‐industrial by‐products to lactic acid using Lactobacillus sakei and two Pediococcus spp. strains

The aim of this study was to evaluate the bioconversion efficiency of rich in cellulose agro‐industrial by‐products such as wheat bran (WB), spent distiller's grain with solids (DGS), brewer's spent grain (BSG) and lupin (Lupinus angustifolius L.) wholemeal fraction (LF) to lactic acid (LA) using acid tolerant lactic acid bacteria (LAB) strains Lactobacillus sakei KTU05‐06, Pediococcus acidilactici KTU05‐7 and P. pentosaceus KTU05‐9. Carbohydrase preparation Depol^? 692L was used for the hydrolysis of non‐starch polysaccharides. Analysed raw materials were suitable substrates for LAB propagation and L‐lactic acid production. The lowest pH (3.6) was found in LF medium after 48 h fermentation with P. acidilactici and P. pentosaceus strains. The lowest pH (3.86) was measured in WB fermented with L. sakei, and in DGS and BSG (pH 3.8 and 3.9 respectively) fermented with P. acidilactici. The highest endoxylanase activity was excreted by the P. acidilactici and P. pentosaceus (84 and 69 XU g^?1 respectively), and the highest α‐amylase activity was of L. sakei (255.6 AU g^?1) after 24 h incubation in WB medium. The L‐lactic acid concentration of 86.11 g kg^?1 was reached after the bioconversion of hydrolysed WB in combination with 48 h fermentation by P. pentosaceus KTU05‐9 strain. LA contents between 222 and 282 mg kg^?1 was produced from lupin processing residues via fermentation using P. acidilactici and P. pentosaceus KTU05‐9 strains. The major challenge within the presented study is the viability of tested LAB in cereal waste media and effective LA production at a low pH (3.6–3.8).     

Nutraceuticals in gummy candies form prepared from lacto‐fermented lupine protein concentrates,as high‐quality protein source,incorporated with Citrus paradise L. essential oil and xylitol

As an addition to a normal diet, the use of food enriched with functional compounds has become increasingly popular. Alternatively, functional ingredients can be incorporated into nutraceuticals in gummy candies (GC) form. This study aimed to adapt lacto‐fermented lupine protein concentrates, as high‐quality protein and genistein sources, with Citrus paradise essential oil (EO), an antimicrobial and bean taste masking agent, and xylitol, a low glycaemic index sweetener, for preparation of nutraceuticals in GC form. For GC preparation, submerged fermentation with Lactobacillus sakei lupine variety Vilniai protein concentrate was selected, which had the highest protein content (90.11%), protein digestibility (89.94%) and genistein content (30.93 μg g^?1) and the lowest trypsin inhibitors activity (19.40%), among the concentrates assessed. GC formulated with xylitol, ascorbic acid, grapefruit EO (up to 0.2%) and lupine protein concentrate (up to 13.0%) allowed obtaining products that exhibited a good texture, high overall acceptability and contained desirable functional compounds.     

Technology and characterisation of whole hemp seed beverages prepared from ultrasonicated and fermented whole seed paste

The aim of this study was to apply fermentation with Pediococcus acidilactici LUHS29, P. pentosaceus LUHS183, Lactobacillus casei LUHS210 and L. uvarum LUHS245 strains, as well as ultrasonication (US) for whole hemp (Cannabis sativa L.) seed paste (HSP) treatment and preparation of emulsions (HSE). Additionally, the biogenic amine (BA) content and antimicrobial properties of the prepared products were evaluated. It was found that US and fermentation reduced total bacteria count in HSP. The highest BA content was found in US-treated HSP (639.87 mg kg⁻¹). HSE_US, HSE_LUHS29, HSE_LUHS183, HSE_LUHS210 and HSE_LUHS245 samples showed lower ζ-potential. The highest overall acceptability (OA) was established for HSE_LUHS245, and correlations were established between the emotions induced by the different HSE products and OA. Pure LAB showed a broad variety of pathogen inhibition; however, the antimicrobial activity of HSP was very low, and HSE did not show any antimicrobial properties. Finally, treatment with selected LAB can be recommended for preparation of stable emulsions, and the most acceptable beverages can be obtained by using LUHS245 strain.     

Development of antimicrobial gummy candies with addition of bovine colostrum,essential oils and probiotics

The aim of this study was to develop antimicrobial properties of gummy candies based on bovine colostrum (BC), essential oils (EOs), lactic acid bacteria (LAB) strains and their combinations. In addition, the heteropolysaccharide (agar), as a multifunctional polymer, was used for the antimicrobial candies preparation. The antimicrobial activities of BC, EOs (C. reticulata L., Eugenia caryophyllata, C. paradisi L., Thymus vulgaris), LAB strains (Lactobacillus plantarum LUHS135 and Lactobacillus paracasei LUHS244) and their combinations against pathogenic bacteria (Pseudomonas aeruginosa, Proteus mirabilis, Escherichia coli, Salmonella enterica, Staphylococcus aureus, Enterococcus faecalis, Streptococcus mutans) were investigated. The highest antimicrobial activities were demonstrated by Thymus vulgaris and Eugenia caryophyllata EOs and their emulsions (12%), and the best formulation of components for antimicrobial gummy candies production would incorporate the BC fermented with L. paracasei LUHS244 in combination with Thymus vulgaris or Eugenia caryophyllata EOs, which inhibited growth of all the tested pathogenic microorganisms (except Pseudomonas aeruginosa). Gummy candies formula consisting of the fermented BC (up to 3%) and thyme EO (up to 0.2%) with mandarin or grapefruit EOs (up to 0.2%) for taste‐masking, allowed obtaining good texture and high overall acceptability products containing desirable antimicrobials, thus antimicrobial gummy candies could be consumer preferred form of nutraceuticals.     

The effects of ultrasonication,fermentation with Lactobacillus sp., and dehydration on the chemical composition and microbial contamination of bovine colostrum

The aim of this study was to evaluate the influence of ultrasonication, fermentation with Lactobacillus plantarum LUHS135 and Lactobacillus paracasei LUHS244, and different methods of dehydration on the chemical composition of bovine colostrum (BC), including the fatty acid and free amino acid profile and the content of micro- and macroelements. In addition, we analyzed the changes in lactic acid bacteria count, microbial contamination (aerobic mesophilic spore-forming bacteria, enterobacteria including Escherichia coli, and fungi/yeasts), the abundance of biogenic amines, and the concentration of nucleotide monophosphates. Significant effects of different treatments on the free amino acid profile were established, and an increase of lysine concentration by 1.2 to 95.9% was observed in treated BC. All of the treatments reduced the concentration of cadaverine, histamine, and tyramine in BC. The concentrations of macro- and microelements in BC followed the following order Ca > Na > K > Mg and Zn > Fe > Sr > Ba > Mn > Cu > Al > Se > Mo > Cr > Ni > Sn > Co > Pb > Cd. By combining the fermentation with Lactobacillus plantarum strain LUHS135 and vacuum drying, it was possible to increase the abundance of nucleotide monophosphates by more than 100%. All of the treatments reduced the microbial contamination of BC. Thus, the combination of ultrasonication, fermentation, and dehydration can be used for improving the properties and safety of BC.     

Evaluation of the use of lactic acid bacteria and Thymus vulgaris essential oil on Suffolk and Ile de France lamb breed (MuscuIus gluteus) quality parameters

In the present study, Lactobacillus plantarum (LUHS135), Thymus vulgaris essential oil (0.1% v/v) and their combination were used to treat Suffolk and Ile de France lamb meat. Changes in the microbiological profile and physicochemical parameters of meat were evaluated after 24 h of treatment at 4 °C. Treatments resulted in significantly lower mould/yeast, total enterobacteria (by up to 45%) counts, water-holding capacity (by 21–63%) and cooking loss (by 17–35%) in all samples. Moisture content increased, on average, by 14.5% in the Suffolk meat. Polyunsaturated fatty acid content increased after all treatments. Treatments and breed significantly affected the content of malondialdehyde and some biogenic amines. Sensory analysis showed that raw lamb meat colour was more acceptable after all treatments, while the odour acceptability was ranked highest after combined treatment. These findings suggest that the applied treatments could be used to improve microbiological safety and some quality characteristics and to increase the PUFA content of lamb meat.