The gene expression profiles of canine mammary cancer cells grown with carcinoma-associated fibroblasts (CAFs) as a co-culture in vitro

<p>Abstract</p> <p>Background</p> <p>It is supposed that fibroblasts present in tumour microenvironment increase cancer invasiveness and its ability to metastasize but the mechanisms have not been clearly defined yet. Thus, the current study was designed to assess changes in gene expression in five various cancer cell lines grown as a co-culture with the carcinoma-associated fibroblasts (CAFs) <it>in vitro</it>.</p> <p>Results</p> <p>A carcinoma-associated fibroblast cell line was isolated from a canine mammary cancer. Then, a co-culture of cancer cells with the CAFs was established and maintained for 72 hrs. Having sorted the cells, a global gene expression in cancer cells using DNA microarrays was examined. The analysis revealed an up-regulation of 100 genes and a down-regulation of 106 genes in the cancer cells grown as a co-culture with the CAFs in comparison to control conditions. The PANTHER binomial statistics tool was applied to determine statistically over-manifested pathways (<it>p </it>< 0.05). Bulk of the up-regulated genes are involved in the adhesion, the angiogenesis, the epithelial-mesenchymal transition (EMT) and generally take part in the developmental processes. These results were further confirmed using real-time qPCR. Moreover, a wound-healing assay and growth characteristics on Matrigel matrix showed that CAFs increase cancer cell migration and matrix invasion.</p> <p>Conclusion</p> <p>The results of the current study showed that the co-culturing of cancer cells and the CAFs caused significant changes to the cancer gene expression. The presence of the CAFs in a microenvironment of cancer cells promotes adhesion, angiogenesis and EMT.</p

CSF-1R as an inhibitor of apoptosis and promoter of proliferation, migration and invasion of canine mammary cancer cells

BACKGROUND: Tumor-associated macrophages (TAMs) have high impact on the cancer development because they can facilitate matrix invasion, angiogenesis, and tumor cell motility. It gives cancer cells the capacity to invade normal tissues and metastasize. The signaling of colony-stimulating factor-1 receptor (CSF-1R) which is an important regulator of proliferation and differentiation of monocytes and macrophages regulates most of the tissue macrophages. However, CSF-1R is expressed also in breast epithelial tissue during some physiological stages i.g.: pregnancy and lactation. Its expression has been also detected in various cancers. Our previous study has showed the expression of CSF-1R in all examined canine mammary tumors. Moreover, it strongly correlated with grade of malignancy and ability to metastasis. This study was therefore designed to characterize the role of CSF-1R in canine mammary cancer cells proliferation, apoptosis, migration, and invasion. As far as we know, the study presented hereby is a pioneering experiment in this field of veterinary medicine. RESULTS: We showed that csf-1r silencing significantly increased apoptosis (Annexin V test), decreased proliferation (measured as Ki67 expression) and decreased migration (“wound healing” assay) of canine mammary cancer cells. Treatment of these cells with CSF-1 caused opposite effect. Moreover, csf-1r knock-down changed growth characteristics of highly invasive cell lines on Matrigel matrix, and significantly decreased the ability of these cells to invade matrix. CSF-1 treatment increased invasion of cancer cells. CONCLUSION: The evidence of the expression and functional role of the CSF-1R in canine mammary cancer cells indicate that CSF-1R targeting may be a good therapeutic approach

Density of Gr1-positive myeloid precursor cells, p-STAT3 expression and gene expression pattern in canine mammary cancer metastasis

The very recent studies on human and mice models have indicated an important role of myeloid precursor cells (progenitors or not fully differentiated cells that express the Gr1 antigen also called Gr1-positive myeloid suppressor cells) in the tumor progression and metastasis. They are thought to suppress the immune system and promote angiogenesis via Signal transducer and activator of transcription 3 (STAT3) activation. As of now there is no data available on the correlation of Gr1-positive cell number, phosphorylated STAT3 (p-STAT3) expression and cancer ability to metastasis. Thus, we counted the myeloid precursor cell number and analyzed p-STAT3 expression in 50 canine mammary tumors that gave local/distant metastases and did not metastasize. We showed that the number of Gr1-positive cells and p-STAT3 expression are significantly higher (p < 0.001) in the metastatic tumors than in the non-metastatic ones. We also observed higher expression of p-STAT3 in the canine mammary cancer cell lines with metastatic potential than in other cell lines (p < 0.001). Moreover, the number of myeloid precursors and p-STAT3 expression in metastatic tumors correlate strongly. The tumor infiltrating myeloid precursor cells may invigorate the STAT3 activity (probably via vascular endothelial growth factor – VEGF) that contributes to the tumor angiogenesis and furthermore tumor`s ability to metastasize. The analysis of gene expression in canine mammary cancer cell lines with metastatic potential indicated that semaphorin 3B (SEMA3B) and neuropilin receptors (NRP) may also be important elements in this process. Thus, we discuss the possible interactions within the tumor that may be required for cancer metastatis

Neurological symptoms in hospitalised patients with COVID-19 and their association with in-hospital mortality

Objectives. To evaluate the spectrum of neurological symptoms in patients with COVID-19 during the first 14 days of hospitalisation and its association with in-hospital mortality. Material and methods. We included 200 patients with RT-PCR-confirmed COVID-19 admitted to University Hospital in Krakow, Poland. In 164 patients, a detailed questionnaire concerning neurological symptoms and signs was performed prospectively within 14 days of hospitalisation. In the remaining 36 patients, such questionnaires were completed retrospectively based on daily observations in the Department of Neurology. Results. During hospitalisation, 169 patients (84.5%) experienced neurological symptoms; the most common were: fatigue (62.5%), decreased mood (45.5%), myalgia (43.5%), and muscle weakness (42.5%). Patients who died during hospitalisation compared to the remainder were older (79 [70.5–88.5] vs. 63.5 [51–77] years, p = 0.001), and more often had decreased level of consciousness (50.0% vs. 9.3%, p &lt; 0.001), delirium (33.3% vs. 4.4%, p &lt; 0.001), arterial hypotension (50.0% vs. 19.6%, p = 0.005) or stroke during (18.8% vs. 3.3%, p = 0.026) or before hospitalisation (50.0% vs. 7.1, p &lt; 0.001), whereas those who survived more often suffered from headache (42.1% vs. 0%, p = 0.012) or decreased mood (51.7% vs. 0%, p = 0.003). Conclusions. Most hospitalised patients with COVID-19 experience neurological symptoms. Decreased level of consciousness, delirium, arterial hypotension, and stroke during or before hospitalisation increase the risk of in-hospital mortality

Regulation of apoptosis: involvement of Bcl-2-related proteins

International audienc

Oględziny i dokumentacja zdarzeń o charakterze upadku z wysokości w terenie eksponowanym i jaskiniach z udziałem ratowników Grupy Jurajskiej Górskiego Ochotniczego Pogotowia Ratunkowego

The article is devoted to issues related to the use of forces and resources of mountain rescue teams during visual inspection of incident areas in exposed areas and caves. The article presents general assumptions about the organization and coordination of this type of inspection from the moment of receiving information about the event and the start of rescue operations by GJ GOPR, until the completion of collecting and securing all infor-mation and evidence as part of the inspection of the incident scene. The article contains issues related to the exchange of information between the involved entities, preparation of documentation meeting formal and legal requirements by trained GJ GOPR rescuers, securing items that may constitute evidence in subsequent proceedings, and the handling of human corpses and remains.The article is devoted to issues related to the use of forces and resources of mountain rescue teams during visual inspection of incident areas in exposed areas and caves. The article presents general assumptions about the organization and coordination of this type of inspection from the moment of receiving information about the event and the start of rescue operations by GJ GOPR, until the completion of collecting and securing all infor-mation and evidence as part of the inspection of the incident scene. The article contains issues related to the exchange of information between the involved entities, preparation of documentation meeting formal and legal requirements by trained GJ GOPR rescuers, securing items that may constitute evidence in subsequent proceedings, and the handling of human corpses and remains

Uproszczona diagnostyka układu napędowego w eksploatacji pojazdu szynowego

Szczególnym przypadkiem niesprawności pojazdów szynowych, często dotyczącym poziomów utrzymania z częściowym demontażem podzespołów, są uszkodzenia elementów układu napędowego, które objawiają się jedynie podczas jazdy. Są one trudne do wykrycia według standardowych metod stacjonarnych. Wykorzystanie metod selekcji negatywnej lub zaawansowanych metod diagnozowania lokalizacji i identyfikacji tych niesprawności, może przyczynić się do znacznego wzrostu kosztów obsługi, które wymiernie wpływają na całkowity koszt eksploatacji pojazdu. W takim przypadku konieczne staje się ich zminimalizowanie, przez określenie jedynie stopnia zakłócenia funkcjonowania poszczególnych elementów układu napędowego pojazdu. Umożliwi to lokalizację obszaru wystąpienia niesprawności i podjęcie dalszych decyzji obsługowych. W artykule przedstawiono wyniki testowego wdrożenia funkcjonalnej diagnostyki kontrolnej układów napędowych w eksploatacji wybranego typu pojazdu szynowego. Przedstawiono wyniki badań eksperymentalnych, wykorzystujących pomiary drgań elementów układu napędowego. Na ich podstawie możliwe jest opracowanie wzorcowych wskaźników zakłócenia funkcjonowania elementów układu napędowego pojazdu do zastosowania w diagnostyce kontrolnej taboru