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Application of Lactobacillus reuteri B1/1 (Limosilactobacillus reuteri) improves immunological profile of the non-carcinogenic porcine-derived enterocytes
Viera Karaffová, Jana Teleky, Maša Pintarič, Tomaž Langerholc, Dagmar Mudroňová, Erik Hudec, Zuzana Ševčíková, 2023, original scientific article

Abstract: In our previous studies, Lactobacillus reuteri B1/1, which was renamed Limosilactobacillus reuteri (L. reuteri), was able to modulate the production of pro-inflammatory cytokines and other components of the innate immune response in vitro and in vivo. In this study, we evaluated the effect of Lactobacillus reuteri B1/1 in two concentrations (1 x 10(7) and 1 x 10(9) CFU) on the metabolic activity, adherence ability and relative gene expression of pro-inflammatory interleukins (IL-1 beta, IL-6, IL-8, IL-18), lumican and olfactomedin 4 produced by non-carcinogenic porcine-derived enterocytes (CLAB). CLAB cells were cultured in a 12-well cell culture plate at a concentration of 4 x 10(5) cells/well in DMEM medium in a controlled humidified atmosphere for 48 h. A 1 mL volume of each probiotic bacterial suspension was added to the CLAB cells. Plates were incubated for 2 h and 4 h. Our results revealed that L. reuteri B1/1 was able to adhere to CLAB cells in sufficient numbers in both concentrations. In particular, the concentration of 109 L. reuteri B1/1 allowed to modulate the gene expression of pro-inflammatory cytokines, as well as to increase the metabolic activity of the cells. In addition, administration of L. reuteri B1/1 in both concentrations significantly stimulated gene expression for both proteins in the CLAB cell line after 4 h of incubation.
Keywords: Lactobacillus reuteri B1/1, CLAB cells, pro-inflammatory cytokine, metabolic activity
Published in DKUM: 20.02.2024; Views: 43; Downloads: 2
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Morphological and immunocytochemical characterization of tumor spheroids in ascites from high-grade serous carcinoma
Simona Miceska, Erik Škof, Gorana Gašljević, Veronika Kloboves-Prevodnik, 2023, original scientific article

Abstract: Tumor spheroids in the ascites of high-grade serous carcinoma (HGSC) are poorly described. Our objective was to describe their morphological features, cellular composition, PD-1 and PD-L1 expression, and survival correlation of these parameters. The density and size of spheroids were assessed in Giemsa-stained smears; the cell composition of spheroids, including tumor cells, immune cells, capillaries, and myofibroblasts, as well as PD-1 and PD-L1 expression on tumor and immune cells was assessed in immunocytochemically stained cell block sections. Forty-seven patients with primary HGSC and malignant ascites were included. A cut-off value for a spheroid density of 10% was established, which significantly predicted overall survival. However, spheroid size did not correlate with survival outcomes. Spheroids were primarily composed of tumor cells, but the presence of lymphocytes and macrophages was also confirmed. Moreover, capillaries were present in the spheroids of three patients, but the presence of myofibroblasts was not confirmed. PD-1 was expressed on lymphocytes but not on tumor cells. PD-L1 expression was seen on both tumor and immune cells, assessed by 22C3 and SP263 antibody clones but not by the SP142 clone. Our results highlight the potential of routine cytopathological techniques to analyze spheroids in HGSC ascites as a valuable tool to investigate their potential as prognostic markers.
Keywords: ascites, cell blocks, high-grade serous carcinoma, ovarian carcinoma, spheroids, spheroid-associated immune cells, PD-1, PD-L1
Published in DKUM: 14.02.2024; Views: 64; Downloads: 3
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Efficiency of laser-shaped photovoltaic cells
Ewa Korzeniewska, Mariusz Tomczyk, Łukasz Pietrzak, Miralem Hadžiselimović, Bojan Štumberger, Klemen Sredenšek, Sebastijan Seme, 2020, original scientific article

Abstract: The main aim of this paper is to analyze the influence of laser shaping of the photovoltaic cell based on its efficiency. The authors described both process of the monocrystalline photovoltaic cell manufacturing, its efficiency, and the possibilities of usage in architecture and the process of creating the photovoltaic cells of unconventional shapes by using laser technology. A method for cutting photovoltaic cells using a fiber laser was presented as well as the parameters of the laser cutting process. The described method allows cutting the massively produced silicon cells according to the predetermined trajectory. Using the proposed process parameters, satisfactory cutting edge quality, and negligible impact of the laser beam on changes in the structure of the photovoltaic cell active layers were achieved. In each cycle of structure cutting, only a small part of the material is removed (from 1 to 2 mikro), and depending on the thickness, the process is repeated from 50 to 300 times. It has been shown that the efficiency of the modified cells depends on the ratio of their surface area to the laser cutting line.
Keywords: photovoltaic cells, efficiency, photovoltaic cell modification, laser modification, silicon cells
Published in DKUM: 15.11.2023; Views: 212; Downloads: 11
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Phenotyping of mammalian CHO cell line in fed-batch bioprocess by intensified and classical procedure : master's thesis
Matic Košir, 2023, master's thesis

Abstract: Nowadays, pharmaceutical industry is striving to find more biologically relevant solutions and to move from synthetic to biological products. Biopharmaceuticals represent an emerging sector in the pharmaceutical industry and are useful for a wide range of indications, including oncology and rheumatology. The development and optimisation of bioprocesses at laboratory scale is crucial for process understanding and translation into production. Bioprocess development often begins with comparison of several potential clones to be characterized, tested in low-volume cultures with different media or other variables, and then evaluated for growth, productivity, and consistency. The purpose of this master's thesis is to review changes in growth curves, substrate consumption, yield, and product quality of a mammalian cell line when substituting glucose as the main carbon source with alternative energy sources. This master thesis presents an attempt to adapt a mammalian CHO cell line to alternative energy sources. A comparison of the cultivation with maintenance of the CHO mammalian cell line by an intensified and a classical process is also shown. Substrate requirements at different time points of the process were monitored using phenotypic microarrays. Based on the obtained and analyzed results of the phenotypic microarrays, we tried to optimize the cell culture supplementation strategy. We have tried to use small molecules to facilitate adaptation to alternative energy sources. Our results showed that growing CHO cell culture on an alternative energy source-maltose and cell reprogramming with small molecules affects the quality of the product. Trends in cell growth, substrate consumption, by-product excretion and final product concentration are similar between the reference cell cultures and those pre-treated with small molecules. We found that adapting a mammalian CHO cell line to an alternative energy source is a challenging process that would need to be carried out over a long period of time.
Keywords: CHO cells, bioprocess, phenotyping, alternative energy sources, glycan mapping
Published in DKUM: 05.10.2023; Views: 182; Downloads: 73
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Phosphonate-modified cellulose nanocrystals potentiate the Th1 polarising capacity of monocyte-derived dendritic cells via GABA-B receptor
Marina Bekić, Miloš Vasiljević, Dušica Stojanović, Vanja Kokol, Dušan Mihajlović, Dragana Vučević, Petar S. Uskoković, Miodrag Čolić, Sergej Tomić, 2022, original scientific article

Abstract: Purpose: Phosphonates, like 3-AminoPropylphosphonic Acid (ApA), possess a great potential for the therapy of bone tumours, and their delivery via cellulose nanocrystals (CNCs) seems a promising approach for their increased efficacy in target tissues. However, the immunological effects of CNC-phosphonates have not been investigated thoroughly. The main aim was to examine how the modification of CNCs with phosphonate affects their immunomodulatory properties in human cells. Methods: Wood-based native (n) CNCs were modified via oxidation (ox-CNCs) and subsequent conjugation with ApA (ApA-CNCs). CNCs were characterised by atomic force microscopy (AFM) and nanoindentation. Cytotoxicity and immunomodulatory potential of CNCs were investigated in cultures of human peripheral blood mononuclear cells (PBMCs) and monocyte-derived dendritic cells (MoDCs)/T cells co-cultures by monitoring phenotype, cytokines production, allostimulatory and Th/Treg polarisation capacity. Results: AFM showed an increase in CNCs’ thickens, elasticity modulus and hardness during the modification with ApA. When applied at non-toxic doses, nCNCs showed a tolerogenic potential upon internalisation by MoDCs, as judged by their increased capacity to up-regulate tolerogenic markers and induce regulatory T cells (Treg), especially when present during the differentiation of MoDCs. In contrast, ox- and ApA-CNCs induced oxidative stress and autophagy in MoDCs, which correlated with their stimulatory effect on the maturation of MoDCs, but also inhibition of MoDCs differentiation. ApA-CNC-treated MoDCs displayed the highest allostimulatory and Th1/CTL polarising activity in co-cultures with T cells. These effects of ApA-CNCs were mediated via GABA-B receptor-induced lowering of cAMP levels in MoDCs, and they could be blocked by GABA-B receptor inhibitor. Moreover, the Th1 polarising and allostimulatory capacity of MoDCs differentiated with ApA-CNC were largely preserved upon the maturation of MoDCs, whereas nCNC- and ox-CNC-differentiated MoDCs displayed an increased tolerogenic potential. Conclusion: The delivery of ApA via CNCs induces potent DC-mediated Th1 polarisation, which could be beneficial in their potential application in tumour therapy.
Keywords: cellulose nanocrystals, phosphonates, dendritic cells, regulatory T-cell subsets, immunomodulation, GABA-B receptor
Published in DKUM: 17.08.2023; Views: 195; Downloads: 14
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The importance of the correct choice of Franz diffusion cell volume for in vitro drug release testing of wound dressings
Tina Maver, Uroš Maver, 2022, original scientific article

Keywords: in vitro drug release testing, Franz diffusion cells, simulation, naproxen, alginate
Published in DKUM: 16.02.2023; Views: 500; Downloads: 38
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Endometrial cancer in young woman
Alenka Repše-Fokter, 2020, short scientific article

Keywords: Pap smear, atypical endometrial cells, young patients
Published in DKUM: 23.01.2023; Views: 503; Downloads: 12
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Suitability of Hydrogen Fuel Cell Powertrain for High-Performance Sports Cars : diplomsko delo
Dominik Ozmec, 2021, undergraduate thesis

Abstract: This bachelor´s thesis describes the development of Hydrogen Fuel Cell powertrain based on the case of the Tushek TS900 hybrid car. We wanted to see if a hydrogen fuel cell-powered vehicle could achieve comparable performance compared to an existing hybrid vehicle. We have built a Fuell Cell propulsion model for the high-performance sports car Tushek TS900, then we presented the basics and principles of the powertrains itself, and its connections. We performed vehicle acceleration simulations and compared them with a hybrid powertrain powered vehicle.
Keywords: hydrogen fuel cells, sports cars, powertrain design, car characteristics, driving simulation
Published in DKUM: 16.02.2022; Views: 585; Downloads: 62
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Saviour siblings - current overview, dilemmas and possible solutions?
Sandra O. Samardžić, 2019, original scientific article

Abstract: In certain cases, when an already born child is affected with a fatal disease and needs transplantation, the only possible solution could be the creation of saviour sibling. Saviour sibling is a term that refers to a child born through a procedure in which an embryo, created in vitro, is being tested in order to determine whether such an embryo could provide stem cells or tissue for an already born, ill child. If the embryo is both, a matching donor and free of the disease, it could be implanted and after the birth of a child, the umbilical cord stem cells or tissue could be used for treatment of a sick sibling. However, this procedure poses a number of dilemmas. This paper aims to give a brief analysis of these issues, to address some of the main concerns and to provide possible solutions for future regulation of this technique.
Keywords: saviour sibling, right of the child to health, medically assisted reproduction, bone marrow, stem cells
Published in DKUM: 15.01.2021; Views: 807; Downloads: 25
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