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Symposium Socratic Lectures is primarily devoted to the students. For 15 years, excellent scientists donate their lectures to the students who are the agens movens, for their dedica-tion and interest. The students were actively involved in discussions. Recently developed technologies enabled creation of documents by simultaneous writing in real time during the symposium. In this contribution, the reflections of the students on the content of symposium are given in terms of statements. Keywords: Education; Open book; Social networks

The gut microbiota is believed to have a major impact on human health and disease. It is involved in barrier functions and maintenance of homeostasis. It provides nutrients and metabolites, participates in a signaling network, regulates epithelial development, and influences the immune system as well as protects the intestinal mucosa from the aggression of pathogenic microorganisms. There is growing evidence that physical activity has an impact on the gut microbiota. Recent studies in animals and humans suggest that regular physical activity increases the presence of beneficial microbial species and improves host health. However, some specific differences should be noted: different forms of physical activity, frequency or intensity, aerobic or resistance training, and benefits and consequences for amateur or competitive athletes. Because the positive role of physical activity can have an impact on health and various types of diseases, the results of research studies in this area are increasingly becoming the focus of scientific interest. In addition, probiotic supplements modulate intestinal microbial flora, and the ability of probiotics to modulate perturbations in immune function after exercise highlights their potential for use in individuals exposed to high levels of physical activity.

Trehalose is a nontoxic disaccharide and a promising cryoprotection agent for medically applicable cells. In this study, the efficiency of combining trehalose with reversible electroporation for cryopreservation of two types of human mesenchymal stromal cells was investigated: adipose-derived stromal cells, and umbilical-cord-derived stromal cells. Comparable results to standard dimethyl sulfoxide cryopreservation protocols were achieved, even without extensive electroporation parameters and protocol optimization. The presence of high extracellular trehalose resulted in comparable cell viabilities without and with electroporation. According to the determination of trehalose concentrations, 250 mM extracellular trehalose resulting in, 20 mM to 50 mM intracellular trehalose were sufficient for successful cryopreservation of cells. With electroporation, higher (i.e. 50 mM to 90 mM) intracellular trehalose was achieved after cryopreservation, although cell survival was not improved significantly. To evaluate the impact of electroporation and cryopreservation on cells, stress and immune-activation-related gene expression were analyzed. Electroporation and/or cryopreservation resulted in increased SOD2 and HSPA1A expression. Despite the increased stress response, the high up-regulation by mesenchymal stromal cells of immunomodulatory genes in the inflammatory environment was not affected. Highest expression was seen for the IDO1 and TSG6 genes. In conclusion, cryopreservation of mesenchymal stromal cells in trehalose results in comparable characteristics to their cryopreservation using dimethyl sulfoxide.

COVID-19 presentations range from cold-like symptoms to severe symptoms with the development of acute respiratory distress syndrome (ARDS). We report on a severe COVID-19 patient who was mechanically ventilated and who developed ARDS and bacterial infection. Because of rapid clinical deterioration and the exhaustion of other treatment options, the family and attending physicians requested a compassionate use of adult allogeneic bone marrow-derived mesenchymal stem cells (MSC) in addition to commonly used immunosuppressive, antiviral, and supportive therapy. The clinical course is discussed thoroughly, with a special emphasis on the safety and effect of MSC therapy. Compassionate MSC treatment, given in three rounds, affected ARDS regression. The patient was discharged from the intensive care unit after 31 days and from hospital after 49 days in a good general condition. MSC treatment was not associated with any side effects and was well tolerated in a three-week period; therefore, it should be studied in larger trials and considered for compassionate use

To document clinical, radiologic, and cellular data of a prospective patient series treated by a tri-layer collagen-hydroxyapatite biomimetic osteochondral scaffold (CHAS) intra-operatively seeded with cultivated autologous chondrocytes (AC) or with filtered bone marrow stem/stromal cells (fBMSC) to address chronic osteochondral knee lesions. Thirty-six consecutive patients (15 to 59 years) with chronic osteochondral lesions (1.8–10 cm2) in the condylar or patellofemoral knee surfaces were enrolled. Lesions were covered with CHAS fixed with a fibrin glue. The superficial layer of CHAS was intra-operatively injected with active cells: in initial five patients, ACs were put directly onto dry CHAS (dry-AC); next, eight AC patients had CHAS moistened with cell culture media (media-AC), while the tourniquet was released allowing blood soaking of CHAS in the rest (14 blood-AC, 9 blood-fBMSC). Seventeen (50%) patients required different concomitant procedures. All patients were followed for serious adverse events (SAE) or graft failures; clinical, radiographic, and MRI evaluation was conducted. Cellular data on the injected cells were assessed. At a follow-up of 39 months (16–81), 17 patients required an additional surgical intervention: seven graft-related SAE (early post-operative synovitis and/or arthrofibrosis) were registered (3 dry-AC, 3 media-AC, 1 blood-fBMSC). There were two graft failures (1 dry-AC, 1 blood-fBMSC) for secondary reasons. All clinical scores significantly improved from pre- to post-operative values: IKCD subjective 44 to 65; IKDC examination (9/17/5/5) to (20/10/5/1); KOOS (P61/S59/ADL67/Sp32/QoL31) to (P79/S75/ADL84/Sp55/QoL51); Tegner activity scale 3.3 to 4.4. There was evidence of radiographic osteoarthritis progression—Kellgren-Lawrence 1.0 to 1.5. MOCART scores at the final follow-up averaged 71 (10 to 95). Graft-type analysis demonstrated an increased rate of graft-related SAE in dry-AC and media-AC, but their final outcomes were equivalent. Cellular data of AC at the implantation were as follows: cells in suspension 9.2 × 106, viability 95%. In blood-fBMSC group, a cell suspension with 87% viability was injected, which contained 1156 CFU-Fs. CHAS with intra-operative seeding of active cells, either AC or fBMSC, led to an overall successful outcome for the treatment of chronic osteochondral lesions in the knee. Blood soaking of CHAS in situ before cell seeding significantly decreased early post-operative adverse events, such as synovitis and arthrofibrosis.

BACKGROUND: Osteochondral injury is a very common orthopaedic pathology, mainly affecting young, active population, with limited current treatment options. Herein we are presenting cellular and early clinical data of a patient series treated for chronic osteochondral lesions in the knee with a filter-based intra-operative bone marrow aspirate (BMA) separation device. METHODS: Fifteen patients with chronic knee osteochondral lesions (60% females, 19–59 years) were included in this prospective case series. Filtered BMA (f-BMA), containing mesenchymal stem/stromal cells (MSCs), was combined with a biomimetic collagen-hydroxyapatite scaffold (CHAS) and implanted into the site of the lesion. Harvested BMA and post-separation f-BMA were analysed for blood cell counts, flow cytometry, and fibroblast colony forming units (CFU-Fs). Patients were followed for serious adverse events and graft failures. Clinical evaluation was assessed using the knee injury and osteoarthritis outcome score (KOOS). In 8 patients a magnetic resonance imaging (MRI)/arthroscopy were performed. RESULTS: Cell suspension contained 0.027% CD271(+) CD45(−) 7-AAD(−) cells, 0.15% CD73(+) CD90(+) CD105(+) cells and 0.0012% CFU-Fs of all nucleated cells with 86% viability. Filtration process resulted in 12.8 (4.0–40.8) fold enrichment in terms of CFU-F content in comparison to initial BMA. No serious adverse events related directly to the osteochondral treatment were reported. After an average follow-up of 20 months (14–25) all KOOS subscales (Symptoms/Pain/Daily activities/Sport and recreation/Quality of life) increased significantly from pre-operative 55/56/67/30/30 to post-operative 73/76/79/51/52 (p values < 0.05), respectively. MRI or arthroscopic evaluation revealed nearly normal to normal overall International Cartilage Repair Society assessment in 7/8 patients. CONCLUSION: The filter-based BMA separation procedure significantly increased the frequency of mesenchymal stem/stromal cells (MSCs), however their concentration was not increased. The clinical evaluation revealed high safety profile of the treatment and resulted in improved clinical status of the patients.

Cryopreservation is the universal technology used to enable long-term storage and continuous availability of cell stocks and tissues for regenerative medicine demands. The main components of standard freezing media are dimethyl sulfoxide (hereinafter Me2SO) and fetal bovine serum (FBS). However, for manufacturing of cells and tissue-engineered products in accordance with the principles of Good Manufacturing Practice (GMP), current considerations in regenerative medicine suggest development of Me2SO- and serum-free biopreservation strategies due to safety concerns over Me2SO-induced side effects and immunogenicity of animal serum. In this work, the effect of electroporation-assisted pre-freeze delivery of sucrose, trehalose and raffinose into human umbilical cord mesenchymal stem cells (hUCMSCs) on their post-thaw survival was investigated. The optimal strength of electric field at 8 pulses with 100 μs duration and 1 Hz pulse repetition frequency was determined to be 1.5 kV/cm from permeabilization (propidium iodide uptake) vs. cell recovery data (resazurin reduction assay). Using sugars as sole cryoprotectants with electroporation, concentration-dependent increase in cell survival was observed. Irrespective of sugar type, the highest cell survival (up to 80%) was achieved at 400 mM extracellular concentration and electroporation. Cell freezing without electroporation yielded significantly lower survival rates. In the optimal scenario, cells were able to attach 24 h after thawing demonstrating characteristic shape and sugar-loaded vacuoles. Application of 10% Me2SO/90% FBS as a positive control provided cell survival exceeding 90%. Next, high glass transition temperatures determined for optimal concentrations of sugars by differential scanning calorimetry (DSC) suggest the possibility to store samples at −80 °C. In summary, using electroporation to incorporate cryoprotective sugars into cells is an effective strategy towards Me2SO- and serum-free cryopreservation and may pave the way for further progress in establishing clinically safe biopreservation strategies for efficient long-term biobanking of cells.

Gut microbiota is the name given today to the bacterial population living in our intestine. It provides nutrients, metabolites and affects the immune system. Recent animals and human studies suggest that regular physical activity increases the presence of beneficial microbial species of gut microbiota and improves the health status of the host. When gut bacteria diversity reduces, there are systemic consequences leading to gastrointestinal, physiological and psychological distress. This review describes the communication pathway of the microbiota-gut-brain axes and other possible mechanisms by which physical activity causes changes in microbiota composition. Furthermore, it provides the latest evidence of the beneficial role of exercise, which in turn can affect health and various disease processes. The results of research studies in this area are increasingly becoming a focus of scientific attention.

Helichrysum italicum (HI) is a Mediterranean plant with well-reported use in traditional medicine for a wide range of applications, including digestive and liver disorders, intestinal parasitic infections, wound healing, stomach ache and asthma. However, little is known about the global mechanism behind its pleiotropic activity.The aim of this study was to explain the mechanism behind the previously demonstrated effects of HI and to justify its use in traditional medicine.A microarray-based transcriptome analysis was used to discover the global transcriptional alterations in primary colon fibroblasts after exposure to HI infusion for 6 h and 24 h. In addition, quantitative real-time PCR was used to verify the microarray results.Altogether we identified 217 differentially expressed genes compared to non-treated cells, and only 8 were common to both treatments. Gene ontology analysis revealed that 24 h treatment with HI infusion altered the expression of genes involved in cytoskeletal rearrangement and cell growth, whereas pathway analysis further showed the importance of interleukin signaling and transcriptional regulation by TP53. For the 6 h treatment only the process of hemostasis appeared in the results of both enrichment analyses. In functional assays, HI infusion increased cell migration and decreased blood clotting and prothrombin time.With the careful evaluation of the role of individual genes, especially SERPING1, ARHGAP1, IL33 and CDKN1A, represented in the enriched pathways and processes, we propose the main mode of HI action, which is wound healing. In addition to its indirect prevention of diseases resulting from the impaired barrier integrity, HI also effects inflammation and metabolic processes directly, as it regulates genes such as LRPPRC, LIPA, ABCA12, PRKAR1A and ANXA6.

Magistrsko delo obravnava sodelovanje zaposlenih pri upravljanju v Radioteleviziji Slovenija, ki predstavlja posebnost v slovenskem prostoru kot edini javni zavod, katerega ustanovni zakon uveljavlja določbe Zakona o sodelovanju delavcev pri upravljanju. Slednji namreč velja le za zaposlene v gospodarstvu, kar postavlja vse preostale zaposlene v neenakopraven položaj, na kar je že večkrat opozorilo tudi Ustavno sodišče. Delo obravnava področji sodelovanja zaposlenih pri upravljanju in delovanja javnih zavodov v Sloveniji in Evropski uniji ter raziskuje in primerja, kako je delavska participacija uveljavljena v Sloveniji, v slovenskih javnih zavodih, na ravni Evropske unije ter v nekaterih evropskih državah, hkrati pa tudi v Radioteleviziji Slovenija in eni izmed nemških televizij. Rezultat preučevanja je ugotovitev, da so zaposleni v slovenskih javnih zavodih kljub izrecni ustavni določbi v neenakopravnem položaju v primerjavi z zaposlenimi v gospodarstvu, saj lahko sodelujejo pri upravljanju le preko svojih predstavnikov v svetu zavoda, ne pa tudi preko svojih predstavnikov v svetu delavcev. Nasprotno se je v tem pogledu Radiotelevizija Slovenije izkazala za pomembno izjemo, ki ne dokazuje samo, da je veljavna zakonodaja na tem področju primerna za vse preostale javne zavode, ampak predvsem uspešno uveljavlja zakonsko določbo, da lahko vodstvo zavoda in svet delavcev razširita področje sodelovanja zaposlenih pri upravljanju, kar je zaposlenim v Radioteleviziji omogočilo celo širši nabor pravic zakonsko določenih za zaposlene v gospodarstvu. Magistrsko delo je prvo delo, ki podrobno obravnava sodelovanje zaposlenih pri upravljanju v Radioteleviziji Sloveniji in lahko pomembno prispeva k argumentaciji pri urejanju zakonodaje na tem področju tudi za vse preostale slovenske javne zavode. The Master thesis addresses the participation of employees in management of The Radio and Television Corporation of Slovenia which is unique in Slovenia as it is the only public institution whose Founding Law enables provisions of the Worker Participation in Management Act. Such conditions usually apply only to employees in the corporate field putting all other public company employees in an unequal position, which has been repeatedly pointed out by the Constitutional Court. The areas of employee participation in the management and operation of public institutions in Slovenia and the European Union are presented in the thesis. The thesis investigates and compares how employee participation is established in Slovenia, Slovenian public institutions, the European Union as a whole and some separate European countries, as well as Radio and Television Corporation of Slovenia and one of the German televisions. The study findings indicate that employees in Slovenian public institutions, despite the explicit constitutional provision, are in an unequal position compared to employees in the corporate field, as public institution employees can only participate in management through their representatives in the council of the institution, but not through their representatives in the works council. In this respect, Radio and Television Corporation of Slovenia proved to be an important exception, which not only proves that the current legislation in this area is suitable for other public institutions, but it also successfully enforced the legal provision that the management of the institution and the works council can expand the scope of employee participation in management, which in return enabled the employees of Radio and Television Corporation of Slovenia to have an even wider range of rights legally determined by the Worker Participation in Management Act than in the corporate field. This Master's thesis is the first work that addresses the participation of employees in the management of Radio and Television Corporation of Slovenia in detail and can significantly contribute to the application of such legislation in this area for other public institutions in Slovenia.