Glucosamine (GlcN) Suppresses the Cytotoxic Activity of NK-92 Cells Against K562 Cells.

<p>(A) Cytotoxicity of NK-92 cells against K562 cells, following treatment of the cultures overnight (ON) with different concentrations of GlcN or <i>N</i>-acetyl GlcN (GlcNAc). (B) Cytotoxicity of NK-92 cells against K562 cells after ON pretreatment with GlcN, followed by 5-mM GlcN supplementation during the assay. All experiments were performed independently five times. Error bars represent standard deviations.</p

Glucosamine (GlcN) Decreases Phosphorylation of FOXO1 and Paxillin in NK-92 Cells.

<p>(A) Representative immunoblots show the immunoprecipitation of whole-cell lysates, using anti-FOXO1 antibodies, followed by immunoblotting with anti-phospho-Ser antibodies (upper), and reblotting with anti-<i>O</i>-GlcNAc (middle) antibodies, of the NK-92 cells untreated or treated with GlcN. (B) Whole-cell lysates were subjected to immunoprecipitation with anti-paxillin antibodies. Immunoprecipitates were analyzed by immunoblotting with anti-phospho-Thr antibodies (upper), and anti- <i>O</i>-GlcNAc (middle). Total paxillin levels were analyzed using the anti-paxillin antibodies (lower).</p

Glucosamine (GlcN) Triggers Nuclear Translocation of Phosphorylated ERK.

<p>(A) Representative images of phospho-ERK (green) expression in NK-92 cells treated with either GlcN and IL2 or IL2 alone. DAPI was used to stain the NK-92 cell nuclei. The merged image shows the overlapping DAPI and phospho-ERK signals. (B) Ratios of the nuclear and cytoplasmic concentrations of p-ERK in the GlcN-treated and untreated NK-92 cells. Sixty individual cells were analyzed in three independent microscopic slides. (C) Imunoblot shows the amount of p-ERK in the nuclei and cytoplasm of control and IL-2 activated (for 60 minutes) cells, treated with GlcN or left untreated. Histone H3 and GAPDH are showing that nuclear and cytosolic fractions were clear.</p

Glucosamine (GlcN) Treatment Affects Cathepsin C and E Intracellular Levels and Localization in NK-92 cells.

<p>(A) Activities of specific cysteine cathepsins (substrate, Z-Phe-Arg-AMC), cathepsin E (KYS-1), and cathepsin C (H-Gly-Phe-AMC) determined in NK-92 cells treated with or without GlcN. Enzyme activities are presented as fold changes relative to the activity of these enzymes in the untreated cells. (B) Secreted cathepsin C activity following GlcN treatment. Results are presented as fold changes relative to the values obtained for the controls samples. Error bars represent standard deviations of the results obtained from five independent experiments. (C) Cathepsin E and C levels in the lysate of the untreated and GlcN-treated NK-92 cells. β-actin levels were used for normalization. (D) Cathepsin C <i>N</i>-glycosylation type in the GlcN-treated and untreated cells determined using EndoH (EH) and PNGaseF (P). (E) Subcellular localization of cathepsins C (white) and E (green) in NK-92 cells cultured with K562 cells, untreated or treated with GlcN. Cell nuclei were stained with DAPI, the right panel represent bright- filter (BF). Merged images show the overlapping signals. (F) Cathepsin C (green) and perforin (red) colocalization in GlcN-treated and untreated cells. The colocalized area panel shows colocalized area calculated by the LAS AF software.</p

Glucosamine (GlcN) Prolongs ERK Phosphorylation in NK-92 cells.

<p>(A) Representative western blots showing the expression of phosphorylated P38, JNK, and ERK, and total MAPK protein levels in NK-92 cells treated with IL2 alone or in combination with GlcN. (B) Quantification of phosphorylated P38, JNK, and ERK levels in cells treated with IL2 alone or in combination with GlcN, normalized to the total protein levels. Error bars represent standard deviations.</p

Glucosamine (GlcN) Prevents Lytic Granule Polarization in NK Cells.

<p>(A) Polarization of granules expressing perforin (green) in the NK-92 and K562 cell conjugates, in presence or absence of GlcN (upper panels). Representative images of at least 60 NK-92/K562 conjugates are shown. Polarization of granules expressing perforin (green) in the mouse primary NK cell and 4T1 cell conjugates, in presence or absence of GlcN (lower panels). Representative images of at least 20 NK/4T1 conjugates are presented. Conjugates were considered polarized when the perforin signal was located in the quarter of the NK cell nearest to the target cell (merged image). Right, bright-filter images, for detection of the conjugates. (B) Distribution analysis, showing the percentage of NK-92 and K562 conjugates with polarized lytic granules in the untreated and GlcN-treated cells. (C) Western blots of NK-92 cell lysates untreated and treated with GlcN, showing the amount of perforin.</p

VALUE CHAIN GENERATOR

&lt;p&gt;The circular bioeconomy can significantly reduce waste and lower the need for mining of primary new materials, but many companies are unable to find and connect with businesses that can utilize their bioresources. In this context, VCG.ai plays a vital role. The Value Chain Generator (VCG) is a digital solution that matches companies based on their resources, including waste, within the scope of the circular bioeconomy. Leveraging artificial intelligence, big data, and machine learning, VCG empowers clusters, business support organizations, and companies to explore and develop new value chain designs, fostering knowledge transfer and enhancing overall knowledge in this economic field. It also connects stakeholders in the regions and beyond.&lt;/p&gt;&lt;p&gt;VCG finds unexploited business opportunities for bio-based value chains based on available data to overcome the existing information gaps. It matches companies according to their resources, including waste, and on established good practices in value chain design. New value chain designs are done by matching production inputs and outputs across industrial sectors. VCG also provides a knowledge transfer network by facilitating the sharing of good (bio)links with other clusters and organisations to increase the overall knowledge. The good practices shared within the system can be applied to individual companies, enabling them to find the right partners to implement selected circular value chain models.&lt;/p&gt;&lt;p&gt;For example, in a new circular business model, waste, particularly bio-waste, can be transformed into bio-plastics through a process of bioconversion or chemical recycling, offering a sustainable alternative to traditional, petroleum-based plastics. This approach not only gives a second life to waste materials, reducing landfill and environmental pollution, but also aids in the creation of a circular economy, where waste is viewed as a valuable resource not as an endpoint.&nbsp;&lt;/p&gt

Viscosity of plasma as a key factor in assessment of extracellular vesicles by light scattering

Extracellular vesicles (EVs) isolated from biological samples are a promising material for use in medicine and technology. However, the assessment methods that would yield repeatable concentrations, sizes and compositions of the harvested material are missing. A plausible model for the description of EV isolates has not been developed. Furthermore, the identity and genesis of EVs are still obscure and the relevant parameters have not yet been identified. The purpose of this work is to better understand the mechanisms taking place during harvesting of EVs, in particular the role of viscosity of EV suspension. The EVs were harvested from blood plasma by repeated centrifugation and washing of samples. Their size and shape were assessed by using a combination of static and dynamic light scattering. The average shape parameter of the assessed particles was found to be ρ ~ 1 (0.94–1.1 in exosome standards and 0.7–1.2 in blood plasma and EV isolates), pertaining to spherical shells (spherical vesicles). This study has estimated the value of the viscosity coefficient of the medium in blood plasma to be 1.2 mPa/s. It can be concluded that light scattering could be a plausible method for the assessment of EVs upon considering that EVs are a dynamic material with a transient identity

Applicability of platelet- and extracellular vesicle-rich plasma in medicine

Platelet-rich plasma is a blood-derived product with proven favourable effects after a local application in various healing disorders. It is also rich with extracellular vesicles - a heterogeneous group of nano- to micro-sized membranous structures -that are considered as the main mediators of regenerative effects. Hence, the prepared blood product can be suitably named »platelet- and extracellular vesicle-rich plasma«. Platelets and platelet- derived extracellular vesicles are not only important in haemostasis, but also in the immune response. Platelets are the most numerous blood immune cells. They are also the main source of blood-derived extracellular vesicles. Extracellular vesicles play an important role in intra- and intercellular communication, therefore they can be utilised in diagnosis and treatment. Platelet- and extracellular vesicle-rich plasma is being used for almost three decades in different fields of medicine, especially in surgery, due to its favourable regenerative properties. However, extracellular vesicles are seldom described in clinical studies that consider the platelet-rich plasma. Based on the molecular mechanisms of the healing process, functions of platelets and platelet-derived extracellular vesicles, platelet- and extracellular vesicle-rich plasma offers an important therapeutic solution in different diseases. Application of platelet- and extracellular vesicle-rich plasma is inexpensive and safe, however its preparation requires advanced laboratory skills. An article contains a description of this blood product and reported experiences on its use. We also present our recent advances which are a product of a collaboration of researchers from medical and biomedical fields. This collaboration leads to an advancement in the treatment modalities in different fields of medicine, also otorhinolaryngology and cervicofacial surgery