Comparison of Cytokine Hemadsorption as an Immunomodulator Therapy in COVID-19 Patients with and without Bacterial Sepsis.

BACKGROUND: In this retrospective study, we aimed to compare the laboratory and clinical results of cytokine hem-adsorption as an immunomodulation therapy in COVID-19 ICU patients with or without sepsis. METHODS: The levels of PCT, CRP, and ferritin were determined as indicators of infection/sepsis; the levels of in-terleukins (IL-6, IL-8 and IL-10, and TNF-α) were determined as indicators of cytokine storm were compared. APACHE score, SOFA score, and mortality rates were compared for the progression of the disease in 23 COVID-19 patients. RESULTS: The therapy was generally successful in reducing the levels of IL-6, IL-8, IL-10, and TNF-α but the levels measured after the procedure did not differ among the patients with or without sepsis, suggesting that the presence of sepsis did not affect the efficacy and function of the cytokine hemadsorption procedure in COVID-19 patients. All parameters were reduced after the procedure except the levels of PCT and ferritin and mortality rates of patients diagnosed with sepsis. The level of PCT was significantly higher in these patients compared with the patients without sepsis while the ferritin and mortality did not show any significant difference between the two groups, suggesting that the cytokine hemadsorption may be safe in the treatment of critical COVID-19 patients. CONCLUSIONS: As a result, the progression of sepsis in COVID-19 may be avoided with cytokine hemadsorption applied as an immunomodulator therapy. However, this therapy should be further explored and validated prior to its introduction to everyday clinical practice when the epidemic conditions end.

Effects of somatostatin, curcumin, and quercetin on the fatty acid profile of breast cancer cell membranes.

Breast cancer is a worldwide commonly found malignancy in women and effective treatment is regarded as a huge clinical challenge even in the presence of several treatment options. Extensive literature is available demonstrating polyphenols as phytopharmaceutical anticancer agents. Among the polyphenols, quercetin and curcumin have been reported to have a strong potential against breast cancer. However, so far, no comprehensive study has been performed to demonstrate the anticarcinogenic effects of curcumin, quercetin, and their combinations with somatostatin on the fatty acid profile of breast cancer cell membranes. We used MCF-7 and MDA-MB231 breast cancer cells incubated with curcumin and quercetin for 24 h, in the absence and presence of somatostatin, at their EC50 concentrations to evaluate membrane fatty acid based functional lipidomics together with the followup of EGFR and MAPK signaling pathways. The two cell lines gave different membrane free fatty acid reorganization. In MCF-7 cells, the following changes were observed: an increase of ω6 linoleic acid in the cells incubated with somatostatin + quercetin and quercetin and a decrease of ω3 acids in the cells incubated with somatostatin + curcumin compared to somatostatin and significant increases of monounsaturated fatty acid (MUFA), mono-trans arachidonic acid levels and docosapentaenoic acid for the cells incubated with somatostatin + quercetin compared to the control cells. In MDA-MB231 cells, incubations with curcumin, quercetin, and somatostatin + quercetin induced the most significant membrane remodeling with the increase of stearic acid, diminution of ω6 linoleic, arachidonic acids, and ω3 (docosapentaenoic and docosahexaenoic acids). Distinct signaling pathway changes were found for these cell lines. In MCF-7 cells, separate or combined incubations with somatostatin and quercetin, significantly decreased EGFR and incubation with curcumin decreased MAPK signaling. In MDA-MB231 cells, incubation with curcumin decreased AKT1 and p-AKT1 (Thr308) levels. Incubation with curcumin and quercetin decreased the EGFR levels. Our results showed that cytostatic and antioxidant treatments can be combined to induce membrane fatty acid changes, including lipid isomerization as specific free radical driven process, and to influence signaling pathways. This study aimed to contribute to the literature on these antioxidants in the treatment of breast cancer to clarify the effects and mechanisms in combination with somatostatin.

Increased plasma soluble tumor necrosis factor receptor-1 and myeloperoxidase activity in patients with obstructive sleep apnea syndrome.

PURPOSE/AIM: Obstructive sleep apnea (OSA) is characterized by recurrent respiratory disorders associated with increased cardiovascular morbidity and mortality. The increment of systemic inflammation in OSA has been considered as the major pathogenic mechanism leading to cardiovascular diseases. There is limited and conflicting information in the literature investigating myeloperoxidase (MPO) activity and soluble tumor necrosis factor receptor-1 (sTNF-R1) levels in OSA patients. The aim of our study is to assess the clinical utility of plasma MPO activity and sTNF-R1 levels as risk markers for systemic inflammation and development of cardiovascular diseases in OSA patients. MATERIALS AND METHODS: 59 OSA patients diagnosed with polysomnograhpy for Apnea-Hypopnea index (AHI), and 26 healthy volunteers enrolled into the study. Plasma MPO activity was measured using a spectrophotometric method. An enzyme-linked immunosorbent assay (ELISA) method was used to detect plasma sTNF-R1 levels. RESULTS: Plasma MPO activity and sTNF-R1 levels were significantly higher (43.2 ± 21.65 vs. 30.44 ± 8.05 p = .0046; 2.379 ± 1.2 vs. 1.086 ± 0.86 p < .0001, respectively) in the total OSA patients compared to the control group. There was a significant weak correlation between MPO activity and disease severity indicator AHI (p = .03 r = .27). CONCLUSIONS: Elevated plasma MPO activity and sTNF-R1 levels in the OSA patients indicate increased systemic inflammation and oxidative stress which might contribute to the higher incidence of cardiovascular diseases. Therefore, we recommend measurement of plasma MPO activity and sTNF-R1 levels in the OSA patients as potential risk predictors for cardiovascular diseases.

Advanced oxidation protein products and ischaemia-modified albumin in obstructive sleep apnea.

BACKGROUND: Several studies have shown that obstructive sleep apnea increases incidence of cardiovascular morbidity and mortality. The high systemic oxidative stress in obstructive sleep apnea has been considered as a major pathogenic mechanism leading to cardiovascular disease. Oxidative stress-related lipid and DNA oxidation in obstructive sleep apnea have been reported in the previous studies. In contrast, there is limited and contradictory information regarding protein oxidation in obstructive sleep apnea patients such as ischaemia-modified albumin and advanced oxidation protein products. Therefore, we aimed to investigate plasma ischaemia-modified albumin and advanced oxidation protein products and their correlation with total oxidative status and total antioxidative capacity in the obstructive sleep apnea patients. METHODS: Plasma ischaemia-modified albumin, advanced oxidation protein products, total oxidative status and total antioxidative capacity were measured in 25 healthy volunteers and 59 obstructive sleep apnea patients diagnosed with polysomnography. RESULTS: Plasma total antioxidative capacity was significantly lower (P = 0·012) and total oxidative status was significantly higher (P < 0·001) in the patients compared to the controls demonstrating increased oxidative stress in the patients. Plasma advanced oxidation protein products were significantly higher in the patients than the controls (P = 0·024). Plasma ischaemia-modified albumin levels were not statistically different between the obstructive sleep apnea patients and controls (P = 0·74). CONCLUSIONS: We conclude that high systemic oxidative stress in obstructive sleep apnea is reflected by increased advanced oxidation protein products without causing an increase in ischaemia-modified albumin.

Low serum copeptin levels in patients with obstructive sleep apnea.

PURPOSE: Copeptin, the C-terminal fragment of antidiuretic hormone (ADH), is a new biomarker that has been found to be elevated in several cardiovascular disorders and is related with prognosis. Patients with obstructive sleep apnea demonstrate a tendency to develop coronary and cerebral atherosclerotic disease. Our aim was to investigate copeptin levels in untreated new diagnosed obstructive sleep apnea patients without manifest cardiovascular disorders in order to determine whether copeptin could be used as a biomarker in this group. METHODS: A total of 60 patients with obstructive sleep apnea, diagnosed with polysomnography, and 23 healthy volunteers were enrolled into this study. Blood samples were collected after overnight fasting, and copeptin level was measured with an enzyme immunoassay method. RESULTS: Patients with obstructive sleep apnea had a higher incidence of hypertension and body mass index but lower serum copeptin level (0.48 ± 0.24. vs. 0.64 ± 0.28 ng/ml, p = 0.007) compared with the healthy controls. There was no significant difference regarding to serum copeptin levels between the moderate (n = 13) and severe (n = 47) obstructive sleep apnea patients (0.42 ± 0.18 vs. 0.49 ± 0.26 ng/ml, p = 0.409). CONCLUSIONS: Rather than reflecting a reduced risk for cardiovascular disorders, we consider that reduced copeptin level is related with disturbed ADH secretion in obstructive sleep apnea patients. Therefore, it would not be advisable to measure copeptin levels in obstructive sleep apnea patients to determine cardiovascular risk, while this marker could be valuable to demonstrate impairment in ADH regulation in this patient group.

Investigating the Activity of Indole-2-on Derivative Src Kinase Inhibitors Against Chronic Myeloid Leukemia Cells.

BACKGROUND: Src family tyrosine kinases play a potential role in Bcr-Abl-induced leukemogenesis. Src kinase inhibitors are reported as selective inhibitors of chronic myeloid leukemia. OBJECTIVE: Since Src kinase inhibitors have an inhibitive effect on chronic myeloid leukemia, indole derivatives (C-1, C-2, C-3) previously found as potent inhibitors of Src kinase were tested against chronic myeloid leukemia in this study. METHODS: Cell viability of K562 and R/K562 cells, antiproliferative and antioxidant effects, and inhibition profiles of Bcr-Abl kinase of indole derivatives were determined compared to dasatinib and imatinib. RESULTS: The results showed that compounds affected cell proliferation and decreased the levels of Bcr/Abl. These results confirmed that the antileukemic activity of compounds was related to Bcr/Abl expression. Docking studies also presented that compounds are inhibitors of both Src and Abl kinases. Calculation of drug-like properties showed that compounds could be potential drug candidates. CONCLUSION: Among indole-2-on derivatives, previously identified as Src kinase inhibitors, C-2 has been discovered to be a strong anticancer drug that is active against susceptible and resistant K562 cell lines and induces apoptosis.

Can the Reduction of Cytokines Stop the Progression of Sepsis?

Objective In this study, we aimed to analyze the laboratory and clinical results of cytokine hemadsorption as an immunomodulation therapy in ICU patients diagnosed with sepsis or septic shock. Methods The levels of procalcitonin (PCT) and C-reactive protein (CRP), determined to be indicators of infection/sepsis, and the levels of interleukins (IL-6, IL-8, and IL-10) and tumor necrosis factor α (TNFα), deemed as indicators of the cytokine storm, were compared among 32 patients before and after the hemadsorption procedure. Results The hemadsorption significantly reduced the levels of IL-6, IL-8, IL-10, TNFα, PCT, CRP, Acute Physiology and Chronic Health Evaluation (APACHE) scores, mortality rate, and Sequential Organ Failure Assessment (SOFA) scores (p<0.05). APACHE scores and the mean predicted mortality rate (PMR) of the non-survivors measured before the procedure was significantly higher than those of survivors (p=0.002 for both). IL-10, APACHE scores, and the mortality rates determined before the hemadsorption procedure were deemed significant parameters to predict the mortality among all ICU patients (p<0.05). IL-10 levels ≤125.3 ng/L, APACHE score >30, and PMR >70.33 were significantly associated with the mortality rates of all patients, indicating that these three parameters determined before the hemadsorption may be good predictors of mortality among ICU patients with sepsis. Conclusion The progression of sepsis in ICU patients may be prevented with cytokine hemadsorption applied as an immunomodulator therapy.

Hybrid Compounds & Oxidative Stress Induced Apoptosis in Cancer Therapy.

Elevated Reactive Oxygen Species (ROS) generated by the conventional cancer therapies and the endogenous production of ROS have been observed in various types of cancers. In contrast to the harmful effects of oxidative stress in different pathologies other than cancer, ROS can speed anti-tumorigenic signaling and cause apoptosis of tumor cells via oxidative stress as demonstrated in several studies. The primary actions of antioxidants in cells are to provide a redox balance between reduction-oxidation reactions. Antioxidants in tumor cells can scavenge excess ROS, causing resistance to ROS induced apoptosis. Various chemotherapeutic drugs, in their clinical use, have evoked drug resistance and serious side effects. Consequently, drugs having single-targets are not able to provide an effective cancer therapy. Recently, developed hybrid anticancer drugs promise great therapeutic advantages due to their capacity to overcome the limitations encountered with conventional chemotherapeutic agents. Hybrid compounds have advantages in comparison to the single cancer drugs which have usually low solubility, adverse side effects, and drug resistance. This review addresses two important treatments strategies in cancer therapy: oxidative stress induced apoptosis and hybrid anticancer drugs.

Effects of Somatostatin and Vitamin C on the Fatty Acid Profile of Breast Cancer Cell Membranes.

BACKGROUND: Vitamin C (Vit C) is an important physiological antioxidant with growing applications in cancer. Somatostatin (SST) is a natural peptide with growth inhibitory effect in several mammary cancer models. OBJECTIVE: The combined effects of SST and Vit C supplementation have never been studied in breast cancer cells so far. METHODS: We used MCF-7 and MDA-MB231 breast cancer cells incubated with SST for 24h, in the absence and presence of Vit C, at their EC50 concentrations, to evaluate membrane fatty acid-profiles together with the follow-up of EGFR and MAPK signaling pathways. RESULTS: The two cell lines gave different membrane reorganization: in MCF-7 cells, decrease of omega-6 linoleic acid and increase of omega-3 fatty acids (Fas) occurred after SST and SST+Vit C incubations, the latter also showing significant increases in MUFA, docosapentaenoic acid and mono-trans arachidonic acid levels. In MDA-MB231 cells, SST+Vit C incubation induced significant membrane remodeling with an increase of stearic acid and mono-trans-linoleic acid isomer, diminution of omega-6 linoleic, arachidonic acid and omega-3 (docosapentaenoic and docosadienoic acids). Distinct signaling pathways in these cell lines were studied: in MCF-7 cells, incubations with SST and Vit C, alone or in combination significantly decreased EGFR and MAPK signaling, whereas in MDA-MB231 cells, SST and Vit C incubations, alone or combined, decreased p- P44/42 MAPK levels, and increased EGFR levels. CONCLUSION: Our results showed that SST and Vit C can be combined to induce membrane fatty acid changes, including lipid isomerization through a specific free radical-driven process, influencing signaling pathways.

Natural Product Inhibitors of Histone Deacetylases as New Anticancer Agents.

Acetylation and deactylation of histones are important determinants of gene expression. Histone deacetylases (HDACs) remove acetyl groups from histones leading to suppression and regulation of epigenetic gene expression. Current studies have demonstrated that HDAC-inhibitors (HDACIs) inducing histone hyperacetylation are promising novel agents in cancer treatment. HDACIs have been shown to have significant anticancer effects with negligible toxicity in the preclinical studies. Ongoing clinial trials are being performed to investigate the efficiency of HDACIs in human cancers. We have reviewed the current knowledge about the molecular mechanisms of action of HDACIs and the outcome of clinical studies using HDACIs in the therapy of several cancers.

Dynamic thiol/disulphide homeostasis before and after radical prostatectomy in patients with prostate cancer.

Thiol groups are important anti-oxidants and essential molecules protecting organism against the harmful effects of reactive oxygen species (ROS). The aim of our study is to evaluate thiol-disulphide homeostasis with a novel recent automated method in patients with localized prostate cancer (PC) before and six months after radical prostatectomy (RP). 18 patients with PC and 17 healthy control subjects were enrolled into the study. Blood samples were collected from the controls subjects and patients before and six months after RP. Thiol-disulphide homeostasis was determined using a recently developed novel method. Prostate-specific antigen (PSA), albumin, total protein, total thiol, native thiol, disulphide and total antioxidant status (TAS) were measured and compared between the groups. Native thiol, total thiol and TAS levels were significantly higher in the control group than the patients before RP (p < .001). There was a non-significant increase in the native thiol, total thiol and TAS levels in the patients six months after RP in comparison to the levels before RP (p values .3, .3 and .09, respectively). We found a significant negative correlation between PSA and thiol levels. Our study demonstrated that the decreased thiol and TAS levels weakened anti-oxidant defence mechanism in the patients with PC as indicated. Increased oxidative stress in prostate cancer patients may cause metabolic disturbance and have a role in the aetiopathogenesis of prostate cancer.

Epoxomicin Sensitizes Resistant Osteosarcoma Cells to TRAIL Induced Apoptosis.

Osteosarcoma (OS) is the second most common primary malign bone neoplasm after multiple myeloma. Despite systemic chemotherapy, OS may give rise to local recurrences and metastases. Resistance to chemotherapy is not rare and is likely to occur in a high number of patients. Novel therapeutic approaches are required in order to efficiently treat osteosarcoma. Tumor necrosis factor (TNF)-related apoptosis inducing ligand (TRAIL) and proteasome inhibitors (epoxomicin, MG132, bortezomib) represent new promising approaches in cancer treatment. The aim of our study is to elucidate the effects of epoxomicin alone or in combination with TRAIL in two TRAIL-resistant OS cell lines, Saos-2 and MG-63 namely. We determined the cytotoxic effects of epoxomicin and/or TRAIL on these two types of OS cells using dimethylthiazolyl 2,5 diphenyltetrazolium bromide (MTT) test and measured apoptosis markers such as pro-apoptotic Bax levels and caspase-3, -8, -9 activities. We used TUNEL assay to demonstrate apoptosis. We investigated dose and time dependent survival rates of OS cells and determined LD50 doses of epoxomicin and TRAIL on OS cell viability after 24, 48, and 72 hour incubations. Concurrent incubation with TRAIL and epoxomicin for 24 hour significantly increased caspase-3, caspase-8, caspase-9 activities and Bax protein levels. Our study demonstrated that the combination of TRAIL with epoxomicin enhances apoptosis, and overcomes TRAIL resistance, denoting promising results for OS therapy in the future.