Inulin-based emulsion-filled gel as a fat replacer in prebiotic- and PUFA-enriched dry fermented sausages

In order to produce fermented sausages with prebiotic fibre and improved fatty acid composition, 16% of pork back fat was replaced with inulin gelled suspension (I) and inulin linseed oil gelled emulsion (IO). Physico-chemical analysis, fatty acid profiles, lipid oxidation, microbiological, textural, colour and sensory analysis were carried out. The fat content was lower in I (31.38%) and IO (35.36%) modified sausages compared to control (44.37%) (P < 0.05). IO sausages had lower SFA and MUFA and higher PUFA content with an improved n-6/n-3 ratio (2.23) (P < 0.05) and α-linolenic acid increment (5.74 g per 100 g). Reformulation led to decrease in springiness, chewiness and hardness and increase in adhesiveness of the sausages. Modified sausages had lower L* and higher a* values, while b* values of I sausages did not differ compared to control sausages. Modified sausages were acceptable regarding all sensory attributes. Lipid oxidation parameters showed higher susceptibility to oxidation and lipolysis in IO sausages.     

Assessing the performance of a spectral reflectance estimation method based on a diffraction grating and a spectroradiometer

Constant improvements in digital cameras have made it possible to use these devices for colour acquisition and reproduction at an advanced level. It is known that a colour match for all observers when changing illumination can only be achieved by matching spectral data. Obtaining spectral data from digital camera RGB values could provide a new way of using the digital camera as a spectrophotometric tool or as a high‐quality colour‐capturing system that is required in more demanding applications. In the present study, two spectral reflectance estimation methods were examined – the Imai–Berns method (ImaiBerns) and the spectral‐sensitivity‐based method (SpecSens). The purpose of the research was to simplify the procedure of the SpecSens method by using a diffraction grating and a spectroradiometer instead of a monochromator, and to compare the results with those of the ImaiBerns approach. Obtained spectral reflectance estimates were evaluated using the root‐mean‐square error and ?E₀₀ metrics. Results of the research show that the ImaiBerns method was superior to SpecSens, most likely because the former method does not require knowledge of the camera spectral sensitivities, which often introduces errors into reflectance estimation calculations. Both methods were successful in predicting black, brownish, and dark patches, as indicated by a low root‐mean‐square error, as well as unsaturated pastel, pink, or skin colours, which produced low ?E₀₀ values. On the other hand, many of the patches with a low root‐mean‐square error also exhibited high ?E₀₀ values, while bright, nearly‐white patches were characterised by a high root‐mean‐square error.     

Differential localization of VE- and N-cadherins in human endothelial cells: VE-cadherin competes with N-cadherin for junctional localization

The two major cadherins of endothelial cells are neural (N)-cadherin and vascular endothelial (VE)- cadherin. Despite similar level of protein expression only VE-cadherin is located at cell-cell contacts, whereas N-cadherin is distributed over the whole cell membrane. Cotransfection of VE-cadherin and N-cadherin in CHO cells resulted in the same distribution as that observed in endothelial cells indicating that the behavior of the two cadherins was not cell specific but related to their structural characteristics. Similar amounts of alpha- and beta-catenins and plakoglobin were associated to VE- and N-cadherins, whereas p120 was higher in the VE-cadherin complex. The presence of VE-cadherin did not affect N-cadherin homotypic adhesive properties or its capacity to localize at junctions when cotransfectants were cocultured with cells transfected with N-cadherin only. To define the molecular domain responsible for the VE-cadherin-dominant activity we prepared a chimeric construct formed by VE-cadherin extracellular region linked to N-cadherin intracellular domain. The chimera lost the capacity to exclude N-cadherin from junctions indicating that the extracellular domain of VE-cadherin alone is not sufficient for the preferential localization of the molecule at the junctions. A truncated mutant of VE-cadherin retaining the full extracellular domain and a short cytoplasmic tail (Arg621-Pro702) lacking the catenin-binding region was able to exclude N-cadherin from junctions. This indicates that the Arg621-Pro702 sequence in the VE-cadherin cytoplasmic tail is required for N-cadherin exclusion from junctions. Competition between cadherins for their clustering at intercellular junctions in the same cell has never been described before. We speculate that, in the endothelium, VE- and N-cadherin play different roles; whereas VE-cadherin mostly promotes the homotypic interaction between endothelial cells, N-cadherin may be responsible for the anchorage of the endothelium to other surrounding cell types expressing N-cadherin such as vascular smooth muscle cells or pericytes.     

Cytotoxicity of some catalysts commonly used in the synthesis of copolymers for biomedical use

The cytotoxicity of four catalysts commonly used for the synthesis of copolymers for biomedical use, such as segmented polyurethanes, was evaluated towards two types of cells, the first being the well-characterized cell line Swiss 3T3 mouse fibroblasts, the second the actual living system that faces any device in contact with blood, i.e. human endothelial cells (HEC). The catalysts were two tertiary aliphatic amines: TMBDA (tetramethylbutanediamine), and DABCO (1–4 diazo (2,2,2) octane); two alkyl tin compounds: DBTDL (dibutyl-tin-dilaurate), and SnOct (stannous octoate). Cytotoxicity tests were carried out by adding to the culture medium, after cell adhesion, different concentrations of each catalyst in dimethylsulphoxide, and keeping them in contact with the monolayer for 72 h. All the catalysts proved to be cytotoxic, although at different extent (in the order: DABCO < TMBDA < SnOct < DBTDL); their dose inhibiting 50% of cell growth (IC50) came out to be lower for 3T3 fibroblasts than for HEC, with the exception of DBTDL, which showed a similar toxicity for both the cell lines. As an example, the cytotoxicity of a polyurethane-amide, laboratory synthesized with DBTDL as catalysts, was checked with fibroblasts. By using both the method of the extract, and that of the direct contact (through a microporous membrane), a moderate to severe cell growth inhibition, related to the Sn content in the material, was observed.     

Effects of Trichoderma harzianum on Photosynthetic Characteristics and Fruit Quality of Tomato Plants

The beneficial role of fungi from the Trichoderma genus and its secondary metabolites in promoting plant growth, uptake and use efficiency of macronutrients and oligo/micro-nutrients, activation of plant secondary metabolism and plant protection from diseases makes it interesting for application in environmentally friendly agriculture. However, the literature data on the effect of Trichoderma inoculation on tomato fruit quality is scarce. Commercially used tomato cultivars were chosen in combination with indigenous Trichodrema species previously characterized on molecular and biochemical level, to investigate the effect of Trichoderma on photosynthetic characteristics and fruit quality of plants grown in organic system of production. Examined cultivars differed in the majority of examined parameters. Response of cultivar Gružanski zlatni to Trichoderma application was more significant. As a consequence of increased epidermal flavonols and decreased chlorophyll, the nitrogen balance index in leaves has decreased, indicating a shift from primary to secondary metabolism. The quality of its fruit was altered in the sense of increased total flavonoids content, decreased starch, increased Bioaccumulation Index (BI) for Fe and Cr, and decreased BI for heavy metals Ni and Pb. Higher expression of swolenin gene in tomato roots of more responsive tomato cultivar indicates better root colonization, which correlates with observed positive effects of Trichodrema.     

N-Terminal Modification of Gly-His-Tagged Proteins with Azidogluconolactone

Site-specific protein modifications are vital for biopharmaceutical drug development. Gluconoylation is a non-enzymatic, post-translational modification of N-terminal HisTags. We report high-yield, site-selective in vitro α-aminoacylation of peptides, glycoproteins, antibodies, and virus-like particles (VLPs) with azidogluconolactone at pH 7.5 in 1 h. Conjugates slowly hydrolyse, but diol-masking with borate esters inhibits reversibility. In an example, we multimerise azidogluconoylated SARS-CoV-2 receptor-binding domain (RBD) onto VLPs via click-chemistry, to give a COVID-19 vaccine. Compared to yeast antigen, HEK-derived RBD was immunologically superior, likely due to observed differences in glycosylation. We show the benefits of ordered over randomly oriented multimeric antigen display, by demonstrating single-shot seroconversion and best virus-neutralizing antibodies. Azidogluconoylation is simple, fast and robust chemistry, and should accelerate research and development.     

Adenosine A1 receptor agonist GR79236 suppresses apnea during all sleep stages in the rat

We tested the hypothesis that N-[(1S, trans)-2-hydroxycyclopentyl]adenosine (GR79236), a novel adenosine A1 receptor agonist, would suppress sleep-related apnea in the rat at doses not associated with hypotension or hypothermia. Nine adult Sprague-Dawley rats were instrumented for chronic recording of sleep by electroencephalographic and electromyographic monitoring. Respirations were measured by single chamber plethysmograph, and blood pressure and heart period were transduced by a telemetric implant. Each rat was polygraphically recorded for 6 hours on four occasions in random order, with recordings for an individual animal separated by at least 3 days. Fifteen minutes prior to each recording (0945 hours) each animal received a 1 ml/kg intraperitoneal bolus injection of one of four injectates: saline (control) or 0.03 mg/kg, 0.3 mg/kg, or 3 mg/kg of GR79236. The study was a repeated-measures balanced design such that each animal was recorded exactly once for each injectate. The rate of spontaneous apneas (pauses > 2.5 seconds) was significantly reduced during all sleep stages by all doses of GR79236. At the highest dose, apnea index was reduced by over 70% in both non-rapid eye movement (NREM) and rapid eye movement (REM) sleep. In contrast, GR79236 had no effect on sleep stage volumes or blood pressure at any dose tested. Heart rate and core temperature were reduced only at the highest dose (3 mg/kg). We conclude that the adenosine A1 receptor agonist GR79236 significantly suppresses apnea expression in all sleep stages at doses not associated with significant changes in sleep architecture, blood pressure, heart rate, or core temperature.