Breast cancer (BC) is one of the leading causes of death in Canadian women, with an average survival rate of 5 years after diagnosis. Early detection of BC can greatly improve patient outcomes and survival. However, a non-invasive BC detection method is not contemporarily available in clinics. Recent studies suggest that proteins in small extracellular vesicles (sEVs) could be promising biomarkers for non-invasive BC early-stage diagnosis. sEVs are membrane-enclosed vesicles secreted by cells, which drive different stages of carcinogenesis in BC. The purpose of this work is to analyse different published proteomics data in order to identify enzymes that can be potentially used as diagnostic biomarkers for BC. For proteomics analyses, sEVs were derived from different metastatic breast cancer cell lines and a non-cancerous epithelial breast cell line. The results were generated from three proteomics approaches: quantitative proteomics, phosphoproteomics, and protein acetylation analysis. The enzymes with high abundances in cancerous cell lines were extracted from the quantitative proteomic data. Similarly, phosphorylated and acetylated enzymes in the cancer cell lines were extracted. Among these approaches, we proposed a list of enzymes, including their metabolic pathways, that can be explored as potential BC biomarkers. Future validation of enzymes using both cancer cell lines and blood from BC patients remains to be determined.

The age-related neurodegenerative diseases drew the interest of the scientific community, due to the gradual increase of the average age in the World’s population. Recent studies indicated the altered cell cycle in the triggering of neurodegenerative diseases such as Alzheimer’s disease (AD). This process seems to involve the nuclear tau, a protein which we previously shown to have a central role in the neuronal in vitro differentiation. In this work, we studied the role of the nuclear tau protein, specifically of the AT8 epitope, in the onset of AD, to evaluate its possible use as an early molecular marker. The immunolocalization in neurons of CA1 region of the human hippocampus from normal, senile and AD subjects, shown that AT8 epitope decreases in senile neurons respect to youngers, indicating its possible role in the ectopic activation of the cell cycle in differentiated cells. Here we show data that improve the knowledge on the role of nuclear tau in neuronal differentiation and cell degeneration in AD, involving the presence/absence of AT8 in the nucleolus of neurons related to a re-entering in the cell cycle. The molecular mechanisms related to the start of AD are not yet clear, so their understanding is very relevant if we consider the social impact of this disease in the human populations.

Understanding the mechanisms of immunosuppression development has formed the basis for modern cancer immunotherapy. However, some immunotherapies, including immune checkpoint inhibitors, have demonstrated limited efficacy in solid tumors. It is noted that additional activation of the immune system after immunotherapy increases the risk of developing autoimmune diseases. Reprogramming T-cell metabolism is critical for enhancing the antitumor immune response. We have developed an approach to reprogramming immune cells by inhibiting the MAPK/ERK pathway through MEKi and the PD-1/PD-L1 immune checkpoint signaling pathway. This approach shown high efficiency on immune cells isolated from mouse bone marrow. We hypothesized that reprogramming of spleen CD8⁺ T-cells could also create a population of immune cells with high antitumor activity. We reprogrammed CD8⁺ T-cells isolated from the spleen of C57BL/6 mice (rsCD8⁺T-cells). The rsCD8⁺T-cells actively migrated into the lungs of mice with Lewis lung carcinoma (LLC) after intravenous injection and in vitro demonstrated high survival, and targeted elimination of cancer stem cell (CSC). In orthotopic LLC model, cell therapy with rsCD8⁺T-cells increased the amount of proliferating CD8⁺ and CD4⁺ T-cells in blood and lung tissue of C57BL/6 mice. In addition, the amount of CSC decreased in the blood and lung of mice treated with rsCD8-T-cells. A morphological study revealed a decrease in the number of metastases in the lung tissue. Thus, reprogramming increases the antitumor activity of spleen CD8⁺ T-cells; the antitumor effects of rsCD8⁺T-cells in vivo are based on the activation of the host immune response by increasing the populations of CD8⁺ and CD4⁺ T-cells and apoptosis of CSCs.

Abstract: Integral membrane proteins, known as Transient Receptor Potential (TRP), channels are cellular sensors for various physical and chemical stimuli in the nervous system, respiratory airways, colon, pancreas, bladder, skin, cardiovascular system, and eyes. TRP channels with their nine subfamilies are classified by sequence similarity, resulting in this superfamily's tremendous physiological functional diversity. Because TRP channels also play essential roles in cellular signaling and allow the host cell to respond to benign or detrimental environmental changes, understanding how each TRP channel responds to its unique forms of activation energy is critical and crucial as its impairment may lead to several diseases, especially carcinogenesis. One of them is pancreatic cancer, one of the deadly types increasing in prevalence without adequate treatment. Scientific research has witnessed steady development in the past few years in understanding the molecular mechanisms that underlie TRP channel disturbance. TRP channels have been reported to be associated with several cancers. This brief review summarizes current knowledge of the molecular role of TRP channels in the development and progression of pancreatic ductal carcinoma to identify potential therapeutic interventions.

Chronic wound resulting from Diabetes Mellitus is a significant cause of amputation worldwide. Secondary infections, lowering of nitric oxide synthase level, reduction of glucose-6-phosphate dehydrogenase levels, improper extracellular matrix remodelling, neuropathy, abnormality of endothelial cell function, and vasculopathy impedes the normal wound healing cycle during diabetes. Multiple studies have concluded that Ser9 phosphorylation causes inhibition of the glycogen synthase kinase-3β (GSK3-β) protein, which is essential for faster diabetic wound healing. Hence this protein could be a potential target for molecular interactions with prospective wound-healing molecules. Verbascoside, martynoside, echinacoside, crenatoside, and salidroside are a few phenylethanoid glycosides that have the potential wound-healing ability by increasing extracellular matrix synthesis, angiogenesis, keratinocyte migration, and the functioning of macrophages and neutrophils. Thus, the five glycosides were subjected to molecular docking with GSK3-β protein (PDB ID: 1I09). This study revealed strong binding interactions with GSK3-β (between -10.2 to -7.3 kcal/mol) and inhibition constants (between 0.032 to 4.397 µM) which suggested potent inhibition of the target protein even at lower concentrations of these compounds. Further, the docked complexes were visualized to find the interaction of the ligands with the amino acid residues. However, further in-vivo and in-vitro studies are required to validate the activity of these phenylpropanoid glycosides in diabetic wound healing.

"Neuroimmunomodulation" defines the modulatory role of the nervous system in relation to immune functions, which reflects part of the bidirectional communication between the nervous system and the immune system. This phenomenon is possible due to the existence of receptors for neurotransmitters (for example norepinephrine or acetylcholine) on immune cells. In fact, these molecules are able to direct and modulate many of the events of the wound healing process. Among these we find nitric oxide (NO): neuromodulator involved in the control of vascular tone and blood pressure, which has a vasodilator and antimicrobial effect. Photodynamic treatments in venous leg ulcers have shown how this treatment stimulates the activity of immune cells involved in healing, which have, among their functions, that of releasing NO into the extracellular space. The experimental results, showed an increase in the expression of iNOS in PDT-treated lesions, underlining its central role in improving the clinical condition of the wound. Furthermore, these results can be associated with the same studies carried out on other neuromediators, confirming the neuroimmunomodulatory role of NO. Since INOS is almost ubiquitous, its over-expression can be observed in almost all tissues: this not only makes it easily detectable, but any treatments based on its capabilities could be valid on many tissues. In the light of such evidence, the versatility of this protein would therefore assume a key role in the definition of new clinical therapies, as well as in the study of the process of wound healing itself.

Environmental pollution impairs cognitive performance and accelerates its associated age decline. Intranasal (IN) administration constitutes a route for xenobiotics delivery to the brain. The main purpose of the present study was to investigate the long-term consequences of IN administration of a chlorpyrifos formulation (fCPF) (10 mg/kg, every other day, for 15-30 days) in mice. We used a set of behavioral tests and evaluated markers of oxidative stress and the activities of enzymes acetylcholinesterase and glutamate transaminases in different brain areas. After 8 months fCPF-free washout period, fur appearance improved, and the body´s injury number decreased in the fCPF group. Scores in locomotion, recognition memory, and anxiety were similar between control and fCPF-treated mice. Notably, improvements in spatial learning and memory were observed in the fCPF group. Several changes in neurochemical markers of oxidative stress and the activities of enzymes from cholinergic and glutamatergic pathways were observed in different brain areas from fCPF-treated mice. Neurochemical disturbances in the intranasally fCPF group were modeled using the human neuroblastoma cell line SH-SY5Y, treated with fCPF and CPF. Here we show that a previous IN exposure to fCPF induces long-term changes in brain biochemistry. These disturbances, acting in concert, could be responsible for the described behavioral observations. Our results emphasize the importance of the IN pathway on the access of xenobiotics to the brain.

Diabetes mellitus (DM) is a chronic metabolic disorder and is associated with impaired wound healing. Non-healing leg and foot ulcers are a frequent significant consequence of diabetes and are caused by a combination of inadequate tissue perfusion, suppression of re-epithelialization, and poor collagen production. Receptor for Advanced Glycation Endproducts (RAGE) is a multiligand cell surface molecule that belongs to the immunoglobulin superfamily is crucial in the pathophysiology of poor wound healing in diabetics. By inhibiting RAGE, a chronic non-healing wound is more likely to undergo angiogenesis, enhance blood supply to hypoxic areas of the wound and decrease in the pro-inflammatory reaction and pro-apoptotic signaling. Phenylethanoid glycosides (PhGs) are a class of natural glycosides, which possesses anti-diabetic, wound healing, antimicrobial, anti-inflammatory, and antioxidant properties. Echinacoside, a phenylethanoid glycoside has a promising role in wound healing by enhancing angiogenesis, promoting keratinocyte migration and proliferation, and enhancing neutrophil and macrophage activity. Consequently, a molecular docking was performed to assess the interaction between Echinacoside and the RAGE receptor (PDB ID: 6VXG). The ligand and receptor had a strong binding interaction, as indicated by the lowest binding energy, which was found to be -6.1 kcal/mol. To further assess the activity of Echinacoside in diabetic wound healing, in-vitro and in-vivo studies are needed.

Lung Cancer is one of the deadliest cancers, responsible for more than 1.80 million deaths annually worldwide, and it is on the priority list of WHO. In the current scenario, when cancer cells become resistant to the drug, making it less effective leaves the patient in vulnerable conditions. To overcome this situation, researchers are constantly working on new drugs and medications that can help fight drug resistance and improve patients’ outcomes. In this study, we have taken five main proteins of lung cancer, namely RSK4 N-terminal kinase, guanylate kinase, cyclin-dependent kinase 2, kinase CK2 holoenzyme, tumour necrosis factor-alpha and screened the prepared Drug Bank library with 1,55,888 compounds against all using three Glide-based docking algorithms namely HTVS, standard precision and extra precise with a docking score ranging from -5.422 to -8.432 Kcal/mol. The poses were filtered with the MM\GBSA calculations, which helped to identify Imidazolidinyl urea C11H16N8O8 (DB14075) as a multitargeted inhibitor for lung cancer, validated with advanced computations like ADMET, interaction pattern fingerprints. Further, it is proposed to optimise the compound with Jaguar and MD Simulation for at least 100ns with NPT ensemble class to analyse the deviation and fluctuations and possible interactions for stability.

Background and aims

Primary biliary cholangitis (PBC) is a slowly progressive cholestatic, autoimmune liver disease, which leads to fibrosis, cirrhosis and liver failure. PBC is characterized by the presence of specific serum antimitochondrial (AMA) and antinuclear (ANA) antibodies. Oxidative stress plays an important role in the pathogenesis of chronic liver diseases. The serum level of 8-isoprostane, a product of lipid peroxidation, is a marker of oxidative stress in vivo. Oxidative stress caused production of interleukin 8 (IL-8), which belongs to pro-inflammatory cytokines. In that way it can cause the recruitment of inflammatory cells and induces a further increase in oxidant stress mediators.

The aim of the present study was to determine whether the degree of lipid peroxidation determined by measuring the serum level of 8-isoprostane influences the progression of PBC. We also evaluated the serum concentration of IL-8 , and studied the correlation between the level of IL-8, 8-isoprostane and specific autoantibodies.

Methods

In the study, 72 PBC patients, 15 pathological controls (patients with other autoimmune liver diseases), and 15 healthy blood donors were enrolled. AMAs and ANAs, Il-8 and 8-isoprostane were detected using commercially-available ELISA kits.

Results

Elevated levels of IL-8 were detected in 58% of patients with PBC. They were significantly higher compared with the control groups: 91.1 ± 20.1 vs. 4.8 ± 0.6 pg/mL, p = 0.0077. In AMA-positive and ANA-positive PBC groups we found that 70% of patients had a higher concentration of IL-8. Serum 8-isoprostane was also significantly elevated in PBC (238.9 [3.8–500.0] pg/mL), in comparison to healthy controls (12.3 [1.6–22.1] pg/mL, p < 0.001). Serum 8-isoprostane values were positively correlated with a higher concentration of IL-8, total cholesterol, bilirubin concentration and severe liver fibrosis (determined by liver biopsy). 8-Isoprostane concentration was not significantly associated to sex, age and biochemical data. Type III procollagen peptide (PIIINP), a serum marker of hepatic fibrogenesis was increased in PBC patients compare to healthy group (p<0.001). Serum 8-isoprostane correlated positively also with PIIINP.

Conclusions

The results show that IL-8 may be an important factor in liver pathologies in patients with PBC, especially in the development of inflammatory processes. A correlation between the concentration of IL-8, 8-isoprostatne and specific autoantibodies was observed. Furthermore, oxidative stress caused by increased lipid peroxidation was determined to be involved in the pathogenesis of PBC. Remarkably, the obtained results indicate that serum 8-isoprostane might be a promising marker for prediction of the degree of liver fibrosis.