Volume 9 (2021): Issue 2 (June 2021)

The mature human placenta and umbilical cord are rich sources of perinatal mesenchymal stem cells (MSCs). Both cell populations have similar characteristics and cellular properties. Each population can differentiate into multiple mesenchymal subpopulations and retain their self-renewal capacities. Perinatal stem cells can be isolated from tissues obtained from the planned cesarean sections and vaginal deliveries. Their isolation is relatively easy, making them readily available for implementation in various in vitro studies and clinical trials. Their differentiation abilities could be used in advanced regenerative medicine protocols to form new bone, cartilage, or tendons. Moreover, their unique anti-inflammatory and immunomodulatory properties have been implemented in the experimental treatment of multiple autoimmune and degenerative diseases. Numerous phase I/II clinical trials confirmed the safety of perinatal MSCs injections and infusions, albeit the efficacy of those cellular therapies should be investigated in the subsequent large-scale randomized trials.

Running title: Clinical applications of the perinatal mesenchymal stem cells

Congenital toxoplasmosis is a rare, non-curable parasite infection, that affects approximately 242 children in Europe each year. Poland has one of the highest rates of congenital toxoplasmosis in Europe. Transmission of Toxoplasma gondii to the fetus results in numerous medical conditions, such as developmental delay, intellectual disabilities, seizures, hearing loss, and blindness. Chorioretinitis is a serious manifestation of congenital toxoplasmosis that can recur even after 25 years from the primary infection, which poses a significant therapeutic challenge. A 41-year-old female reported to the Ophthalmology Emergency Room due to blurred vision and pain in the right eye, accompanied by a constant headache. The patient suffered from congenital toxoplasmosis with two relapses in the past. On examination, the best-corrected visual acuity was 1,0 in both eyes, and the intraocular pressure was significantly increased. Slit-lamp examination showed vitritis and an active retinochoroidal lesion in the right eye. In the left eye, there was a retinochoroidal scar. A relapse of toxoplasmosis was suspected. Serology for Toxoplasma gondii was positive. Pyrimethamine with sulfadiazine, clindamycin, topical corticosteroids, and intraocular pressure-lowering drugs were implemented. During the treatment, the patient developed corticonuclear cataract in both eyes and reported psychotic symptoms. Clinical condition improved after the treatment with corticosteroids at a lower dose. Treatment of ocular manifestations of congenital toxoplasmosis is challenging. The clinical benefit of treatment should be weighed against side effects for each patient.

Running title: Congenital toxoplasmosis treatment

Cell migration is an essential process for wound healing, metastasis and inflammation. Focal adhesions (FA) are local regions of plasma membrane consisting of multiprotein complexes providing adhesive contact between the cell and the extracellular matrix (ECM). FA turnover regulates different signalling pathways implicated in various cellular responses (e.g. cell migration). Endocytosis, specifically the dynamin and clathrin pathways, is known to regulate cell migration by modulating FA dynamics. In this study, we investigated whether NO activity regulates cell migration, FA dynamics and early endosome trafficking in MDA-MB-231 cells. The assessment of cell migration showed a slowing down of cell migration and an increased duration of FA turnover in cells treated with inhibitors of NO synthase (NOS) such as L-NAME or 1400W. In addition, these treatments were found to exhibit no effect on transferrin and dextran uptake mediated by endocytosis and micropinocytosis, respectively. The number of early endosome antigen 1 (EEA1)-positive endosomes was reduced while their sizes were found to increase in cells treated with L-NAME or 1400W. In contrast, these inhibitors did not affect the number nor the size of Rab5-positive endosomes. Furthermore, we demonstrated that EEA1, endothelial NO synthase (eNOS) and inducible NO synthase (iNOS) were colocalised. Using the biotin switch assay followed by western blot, we showed that early endosome proteins such as APPL1, EEA1, Rab5 were found to be S-nitrosylated. These results were further supported by the sequence analysis performed with the GPS-SNO algorithm which predicted the S-nitrosylation of these endosomal proteins. Taken together, our findings suggest that NO might be involved in cell migration and FA turnover through early endosome trafficking in MDA-MB-231 cells.

Running title: Nitric oxide in MDA-MB-231 breast cancer cells

Stem cells are an important subject of research, and are increasingly used in the treatment of various diseases. Due to the development of advanced in vitro techniques, they have become an integral part of modern medicine. The sources of human stem cells are primarily bone marrow and adipose tissue, although non – embryonic stem cells are also scattered throughout the body. Notably, recent research has focused on stem cells found in the oral cavity, both in the dental pulp and oral mucosa. Furthermore, isolation of stem cells from umbilical cord blood is also becoming increasingly popular, while wharton’s jelly and amniotic fluid also seem to be an interesting source of stem cells. The safety and efficacy of stem cells use can be established by animal studies, which are a key element of preclinical research. Mouse, rat and pig models allow for testing of stem cell therapies. Recent studies primarily use mesenchymal stem cells such as mouse – adipose derived mesenchymal stem cells and mouse and rat hematopoietic stem cells. Great hope for future therapies is the use of bioengineering to program cells into induced stem cells, which have the biggest ability for differentiation and transdifferentiation, which carries no risk of teratogenesis. Stem cells are used in many areas of medicine, especially in regenerative medicine, with a growing interest in orthopedics and in the treatment of heart failure. Mesenchymal stem cells are the most used stem cell type, which despite their limited ability to differentiate, give great therapeutic results, mainly due to their immunomodulating effect. Recent studies have even shown that the use of mesenchymal stem cells may be useful in the treatment of COVID-19. Moreover, Research on the use of mesenchymal stem cells in the treatment of Crohn’s disease, acute-graft-versus-host disease and type I diabetes are also promising. The aim of the current review is to present and systematize current knowledge about stem cells, their use and related in vitro research.

Running title: Research and use of human stem cells

The methicillin-resistant Staphylococcus aureus (MRSA) is a significant human pathogenic bacterium that is endemic within hospitals around the world. The identification and inspection of MRSA in clinical samples is quite helpful both in advising individual patients about the required care and in tracking these species. The goal of this study was to present a modern, faster, and more accurate diagnostic technique to operate on the real-time duplex PCR applicable to S. aureus/MRSA monitoring in Iraqi patients. For this reason, the S. aureus-specific nuc gene sequence and the mecA gene sequence were checked simultaneously. To estimate the assay efficiency, a set of six target strains, 34 non-target strains, and 296 clinical specimens were used. The findings obtained from the diagnosis of a total of 296 isolates based on phenotypic characteristics and biochemical tests showed that 146 (49.32%) were classified as individuals with respiratory tract infections of S. aureus with a total male to female ratio of 1.47, and 142 isolates demonstrated methicillin resistance. 142 MRSA isolates were investigated in the molecular analysis, all MRSA isolates had positive results for the nuc gene and 138 isolates were positive for the mecA gene. The current real-time PCR assay has 97% sensitivity, 100% specificity, and 98% accuracy.

Running title: Identification of the MRSA by real time PCR

Immobilization of antibodies has a number of promising applications, including detection of biomolecules and cells. Well-oriented antibodies are required to bind them effectively. To eliminate the problem of random antibodies’ orientation, the surface of the device can be modified with silanes. This study aimed at elucidating if selected aminosilanes were able to bind antibodies in the appropriate orientation and thus retain their binding activity. Silanization of glass slides was performed using three amino-functional trialkoxysilanes – A, AE, and AEE. The immunofluorescent reaction was used to evaluate the potential of the silanized glass surface to bind anti-EpCAM antibodies. The affinity of selected anti-EpCAM HEA125 antibodies labeled with fluorochrome to tested silanized surfaces was evaluated by measuring the mean fluorescence intensity (MFI) in each analyzed area. The presented silanes effectively bound antibodies. Higher fluorescence intensity was noticed in the case of silane-coated glass slides in comparison to unmodified ones. The differences in the contact angles also confirmed this result. In the case of silane A, the fluorescence intensity reflected the amount of bound antibodies. However, there was no such a relation in the case of the silanes AE and AEE. Although our research gave promising results, the usefulness of selected silanes needs to be confirmed by further studies using cancer cells.

Running title: Aminosilanes as enhancers of antibody immobilization