Concentration of Apoptotic Factors in Bronchoalveolar Lavage Fluid, as Potential Brain-Lung Oxygen Relationship, Correspond to the Severity of Brain Injury.

BACKGROUND: The mechanism of acute brain injury initiates a cascade of consequences which can directly cause lung damage, and this can contribute to poor neurological outcomes. The aim of this study was to evaluate concentration of different apoptotic molecules in the bronchoalveolar lavage fluid (BALF) in patients after severe brain injury and to correlate them with selected clinical variables and mortality. METHODS: Patients with brain injury receiving BALF operation were included in the study. BALF samples were collected within the first 6-8 hours after traumatic brain injury (A) and at days 3 (B) and 7 (C) after admission to the intensive care unit (ICU). Changes in the BALF nuclear-encoded protein (Bax), apoptotic regulatory protein (Bcl-2), pro-apoptotic protein (p53) and its upregulated modulator (PUMA), apoptotic protease factor 1 (APAF-1), Bcl-2 associated agonist of cell death (BAD) and caspase-activated DNase (CAD) were analysed. These values were correlated with the selected oxygenation parameters, Rotterdam computed tomography (CT) score, the Glasgow Coma Score and 28-day mortality. RESULTS: We found a significant increase in the concentration of selected apoptotic factors at admission (A), at day 3 (B) and day 7 (C) after severe brain damage contrasted with baseline level A (p < 0.001, separately). That concentration of selected apoptotic factors was significantly correlated with the severity of the injury and mortality. CONCLUSIONS: Activation of different apoptotic pathways seems to be an important process occurring in the lungs of patients in the early phases after severe brain trauma. Levels of apoptotic factors in the BALF correlates with the severity of brain injury.

Humulus lupus extract rich in xanthohumol improves the clinical course in critically ill COVID-19 patients.

BACKGROUND: The systemic inflammatory response following severe COVID-19 is associated with poor outcomes. Several anti-inflammatory medications have been studied in COVID-19 patients. Xanthohumol (Xn), a natural extract from hop cones, possesses strong anti-inflammatory and antioxidative properties. The aim of this study was to analyze the effect of Xn on the inflammatory response and the clinical outcome of COVID-19 patients. METHODS: Adult patients treated for acute respiratory failure (PaO2/FiO2 less than 150) were studied. Patients were randomized into two groups: Xn - patients receiving adjuvant treatment with Xn at a daily dose of 4.5 mg/kg body weight for 7 days, and C - controls. Observations were performed at four time points: immediately after admission to the ICU and on the 3rd, 5th, and 7th days of treatment. The inflammatory response was assessed based on the plasma IL-6 concentration, neutrophil-to-lymphocyte ratio (NLR), platelet-to-lymphocyte ratio (PLR), C-reactive protein (CRP) and D-dimer levels. The mortality rate was determined 28 days after admission to the ICU. RESULTS: Seventy-two patients were eligible for the study, and 50 were included in the final analysis. The mortality rate was significantly lower and the clinical course was shorter in the Xn group than in the control group (20% vs. 48%, p < 0.05, and 9 ± 3 days vs. 22 ± 8 days, p < 0.001). Treatment with Xn decreased the plasma IL-6 concentration (p < 0.01), D-dimer levels (p < 0.05) and NLR (p < 0.01) more significantly than standard treatment alone. CONCLUSION: Adjuvant therapy with Xn appears to be a promising anti-inflammatory treatment in COVID-19 patients.

Mechanical ventilation in neurocritical care setting: A clinical approach.

Neuropatients often require invasive mechanical ventilation (MV). Ideal ventilator settings and respiratory targets in neuro patients are unclear. Current knowledge suggests maintaining protective tidal volumes of 6-8 ml/kg of predicted body weight in neuropatients. This approach may reduce the rate of pulmonary complications, although it cannot be easily applied in a neuro setting due to the need for special care to minimize the risk of secondary brain damage. Additionally, the weaning process from MV is particularly challenging in these patients who cannot control the brain respiratory patterns and protect airways from aspiration. Indeed, extubation failure in neuropatients is very high, while tracheostomy is needed in one-third of the patients. The aim of this manuscript is to review and describe the current management of invasive MV, weaning, and tracheostomy for the main four subpopulations of neuro patients: traumatic brain injury, acute ischemic stroke, subarachnoid hemorrhage, and intracerebral hemorrhage.

Neuroprotective Effect of Subdural Infusion of Serp-1 in Spinal Cord Trauma.

Spinal cord injury (SCI) initiates a severe, destructive inflammation with pro-inflammatory, CD68+/CD163-, phagocytic macrophages infiltrating the area of necrosis and hemorrhage by day 3 and persisting for the next 16 weeks. Inhibition of macrophage infiltration of the site of necrosis that is converted into a cavity of injury (COI) during the first week post-SCI, should limit inflammatory damage, shorten its duration and result in neuroprotection. By sustained subdural infusion we administered Serp-1, a Myxoma virus-derived immunomodulatory protein previously shown to improve neurologic deficits and inhibit macrophage infiltration in the COI in rats with the balloon crush SCI. Firstly, in a 7 day long study, we determined that the optimal dose for macrophage inhibition was 0.2 mg/week. Then, we demonstrated that a continuous subdural infusion of Serp-1 for 8 weeks resulted in consistently accelerated lowering of pro-inflammatory macrophages in the COI and in their almost complete elimination similar to that previously observed at 16 weeks in untreated SCI rats. The macrophage count in the COI is a quantitative test directly related to the severity of destructive inflammation initiated by the SCI. This test has consistently demonstrated anti-inflammatory effect of Serp-1 interpreted as neuroprotection, the first and necessary step in a therapeutic strategy in neurotrauma.

Effectiveness of Haemophilus influenzae type b vaccination after splenectomy - impact on selected immunological parameters.

Splenectomy is a surgery indicated in case of splenic rupture after injury, when there are tumors in the spleen, or as a treatment for certain diseases, such as idiopathic thrombocytopenic purpura and spherocytosis. The aims of the study were to assess the immunological response to the Haemophilus influenzae type b (Hib) vaccine and the post-vaccination changes in lymphocyte subsets and cell activation markers in splenectomized patients and healthy volunteers. Blood samples were collected from 25 patients that had undergone splenectomy and from 15 healthy, non-splenectomized volunteers. All participants received a single dose of Hib vaccine. The concentration of specific Hib antibodies was assessed by an enzyme-linked immunosorbent assay. Selected immune cell populations were evaluated using flow cytometry. The analysis of the antibody titers against Hib showed statistically significant differences in both groups. There was a significantly higher percentage (p = 0.0012) and absolute value (p = 0.0003) of natural killer T (NKT)-like cells (CD3+/CD16+ CD56+) in the study group, compared to the control group. The levels of natural killer (NK) and NKT cells did not change relative to the cause and age of splenectomy. The quantity and percentage of regulatory T (Treg) cells were higher in the study group compared to the control group (p < 0.0001). No significant correlations were found between the time elapsed since splenectomy, the age of the patients, and the Treg levels. Our study showed that spleen resection results in an important deterioration of Treg cells and Th17 cell balance which may contribute to an incomplete immunological response.

Cytotoxicity of Iron (III), Molybdenum (III), and their Mixtures in BALB/3T3 and HepG2 Cells.

INTRODUCTION: Iron and molybdenum are essential trace elements for cell metabolism. They are involved in maintaining proper functions of enzymes, cell proliferation, and metabolism of DNA. MATERIAL AND METHODS: BALB/3T3 and HepG2 cells were incubated with iron chloride or molybdenum trioxide at concentrations from 100 to 1,400 µM. The cells were also incubated in mixtures of iron chloride at 200 µM plus molybdenum trioxide at 1,000 µM or iron chloride at 1,000 µM plus molybdenum trioxide at 200 µM. Cell viability was determined with MTT reduction, LHD release, and NRU tests. RESULTS: A decrease in cell viability was observed after incubating both cell lines with iron chloride or molybdenum trioxide. In cells incubated with mixtures of these trace elements, a decrease in cell viability was observed, assessed by all the used assays. CONCLUSION: Iron (III) and molybdenum (III) decrease cell viability in normal and cancer cells. A synergistic effect of the mixture of these elements was observed.

Cell Viability in Normal Fibroblasts and Liver Cancer Cells After Treatment with Iron (III), Nickel (II), and their Mixture.

INTRODUCTION: Nickel and iron are very commonly occurring metals. Nickel is used in industry, but nowadays it is also used in medical biomaterials. Iron is an element necessary for cell metabolism and is used in diet supplements and biomaterials, whence it may be released along with nickel. MATERIAL AND METHODS: BALB/3T3 and HepG2 cells were incubated with iron chloride or nickel chloride at concentrations ranging from 100 to 1,400 µM. The following mixtures were used: iron chloride 200 µM plus nickel chloride 1,000 µM, or iron chloride 1,000 µM plus nickel chloride 200 µM. The cell viability was determined with MTT, LHD, and NRU tests. RESULTS: A decrease in cell viability was observed after incubating the BALB/3T3 and HepG2 cells with iron chloride or nickel chloride. A synergistic effect was observed after iron chloride 1,000 µM plus nickel chloride 200 µM treatment in all assays. Moreover, the same effect was observed in the pair iron chloride 200 µM plus nickel chloride 1,000 µM in the LDH and NRU assays. CONCLUSIONS: Iron (III) and nickel (II) decrease cell viability. Iron chloride at a concentration of 200 µM protects mitochondria from nickel chloride toxicity.

Subpopulations of natural killer-T-like cells before and after surgical treatment of laryngeal cancer.

INTRODUCTION: Tumours connected with head and neck comprise about 5% of all tumours. The most frequent histological type of laryngeal carcinoma is squamous cell carcinoma. Different research projects suggest that the role of T lymphocytes might be significant in tumour development. iNKT cells are a new subpopulation of T cells and show cytotoxic activity against tumours. iNKT cells participate in modulating the function of other cells which have anti-tumour properties and secrete cytokines, which have pro-inflammatory and anti-inflammatory effects. In animal models the significance of iNKT cells in various diseases including cancer was shown. AIM OF THE STUDY: The aim of this study was to determine the percentages of iNKT cells, CD161+ cells, CD161- cells, iNKT CD4+ cells, and iNKT CD8+ cells, NK cells, NKT-like cells, and T cells subsets present in peripheral blood of patients with laryngeal cancer before and two months after the tumour resection, in comparison to healthy volunteers. MATERIALS AND METHODS: This study included material from laryngeal patients who were treated at the Department of Otolaryngology and Laryngological Oncology (Medical University of Lublin) between 2012 and 2013. A total of 50 patients (40 men and 10 women) aged between 45 and 77 years (median age: 60 years) were enrolled. Based on the TNM classification, the patients were classified as having stage I-IV laryngeal cancer. The control group was composed of 15 healthy volunteers (12 men and three women) aged between 43 and 82 years (median age: 61 years). The protocol of the study was approved by the Local Bioethical Committee at the Medical University of Lublin.Peripheral blood samples (15 ml) from the basilic vein were collected by venipuncture using sterile, sodium heparin-treated tubes (20 units per ml of blood) and used for cytometric analyses. RESULTS: iNKT cells were analysed among T CD3+ cells. The percentage of CD3+ and CD3+CD4+ T cells before tumour resection was higher than in the control group, but the increase of CD3+ T cells was not significant. The T CD3+CD4+ / T CD3+CD8+ cell ratio was significantly higher than in healthy donors. After tumour resection a decreased percentage of CD3+CD4+ T cells but an increased percentage of CD8+CD3+T cells was noted. The T CD3+CD4+ / T CD3+CD8+ cell ratio was significantly higher in patients before and after the surgery than in the control group. The amount of NKT-like cells increased after resection and was significantly higher than in the control group. CONCLUSIONS: Our study exhibited the change in percentage of iNKT, NK, NKT-like cells, and T lymphocytes after tumour resection in patients with laryngeal cancer. The research explains the contribution of those cells in immunological response against tumour.

Myeloid-derived suppressor cells in ovarian cancer: friend or foe?

Although previous decades contributed to major progress in targeted therapy of many malignancies, the treatment of gynaecological cancers remains a challenging task. In the evidence of rising cancer mortality, the search for new methods of treatment is a dire need. Exploring the mechanisms of interaction between tumour cells and host immune response may allow the introduction of new, effective therapies - not as toxic and far more efficient than conventional methods of cancer treatment. Epithelial ovarian cancer (EOC) is typically diagnosed at advanced stages. Its incidence and mortality rate is high. Powerful diagnostic tools for this kind of cancer are still under investigation. Multiple mechanisms existing in the ovarian tumour network create a specific immunosuppressive microenvironment, in which accumulation of myeloid-derived suppressor cells (MDSCs) may be a critical component for diagnosis and treatment. This review attempts to verify current knowledge on the role of MDSCs in EOC.