[The impact of electronic cigarettes usage on the endothelial function and the progression of atherosclerosis]. / Wplyw uzywania papierosów elektronicznych na funkcje sródblonka naczyniowego oraz rozwój miazdzycy.

The exponetial growth in popularity of electronic cigarettes in the world markets intensifies the debate about their health effects. The smoking of traditional tabacoo products is a factor associated with the endothelium damage and progression of atherosclerosis. The elimination of the combustion process in electronic cigarettes allows to conclude that they are less harmful to a vascular endothelium than traditional tobacco products. E-cigarette aerosol contains many compounds that have an influence on initiation and progression of atherosclerosis. Nicotine protherogenic action is not fully explained. On one hand, nicotine modifies metabolic pathways leading to atherosclerosis, whereas epidemiological studies do not show an increased risk of cardiovascular disease in the population using nicotine replacement therapy or snuff. Acrolein, formaldehyde and the ultrafine particles generated during e-liquid heating have an impact on initiation and progression of atherosclerosis, but their level is lower than that of tobacco smoke. In order to assess accurately the longterm effects of e-cigarettes, it is necessary to conduct epidemiological studies measuring the effects of using e-cigarettes. It is claimed that the use of electronic cigarettes has a potential impact on the development of atherosclerosis, but is significantly lower than that of traditional cigarettes.

Extended magnetic resonance imaging studies on the effect of classically activated microglia transplantation on white matter regeneration following spinal cord focal injury in adult rats.

Spinal cord injuries are still a serious problem for regenerative medicine. Previous research has demonstrated that activated microglia accumulate in spinal lesions, influencing the injured tissues in various ways. Therefore, transplantation of activated microglia may have a beneficial role in the regeneration of the nervous system. The present study examined the influence of transplanted activated microglial cells in adult rats with injured spinal cords. Rats were randomly divided into an experimental (M) and control (C) group, and were subjected to non-laminectomy focal injury of spinal cord white matter by means of a high-pressured air stream. In group M, activated cultured microglial cells were injected twice into the site of injury. Functional outcome and morphological features of regeneration were analyzed during a 12-week follow-up. The lesions were characterized by means of magnetic resonance imaging (MRI). Neurons in the brain stem and motor cortex were labeled with FluoroGold (FG). A total of 12 weeks after surgery, spinal cords and brains were collected and subjected to histopathological and immunohistochemical examinations. Lesion sizes in the spinal cord were measured and the number of FG-positive neurons was counted. Rats in group M demonstrated significant improvement of locomotor performance when compared with group C (P<0.05). MRI analysis demonstrated moderate improvement in water diffusion along the spinal cord in the group M following microglia treatment, as compared with group C. The water diffusion perpendicular to the spinal cord in group M was closer to the reference values for a healthy spinal cord than it was in group C. The sizes of lesions were also significantly smaller in group M than in the group C (P<0.05). The number of brain stem and motor cortex FG-positive neurons in group M was significantly higher than in group C. The present study demonstrated that delivery of activated microglia directly into the injured spinal cord gives some positive effects for the regeneration of the white matter.

Exenatide and metformin express their anti-inflammatory effects on human monocytes/macrophages by the attenuation of MAPKs and NFκB signaling.

Metformin and exenatide are effective antidiabetic drugs, and they seem to have pleiotropic properties improving cardiovascular outcomes. Macrophages' phenotype is essential in the development of atherosclerosis, and it can be modified during antidiabetic therapy, resulting in attenuated atherogenesis. The mechanism orchestrating this phenomenon is not fully clear. We examined the impact of exenatide and metformin on the level of TNF alpha, MCP-1, reactive oxygen species (ROS), and the activation of mitogen-activated protein kinases (MAPK), nuclear factor kappa B (NFκB), and CCAAT/enhancer-binding protein beta (C/EBP beta) in human monocytes/macrophages. We found that both drugs reduced levels of TNF alpha, ROS, and NFκB binding activity to a similar extent. Compared to metformin, exenatide was more effective in reducing MCP-1 levels. We noted that Compound C (AMPK inhibitor) reduced the impact of exenatide on cytokines, ROS, and NFκB in cultures. Both drugs elevated the C/EBP beta phosphorylation level. Experiments on MAPKs showed effective inhibitory potential of exenatide toward p38, JNK, and ERK, whereas metformin inhibited JNK and ERK only. Exenatide was more effective in the inhibition of JNK than metformin. Interestingly, an in vitro setting additive effect of drugs was absent. In conclusion, here, we report that metformin and exenatide inhibit the proinflammatory phenotype of human monocytes/macrophages via influence on MAPK, C/EBP beta, and NFκB. Exenatide was more effective than metformin in reducing MCP-1 expression and JNK activity. We also showed that some effects of exenatide relied on AMPK activation. This shed light on the possible mechanisms responsible for pleiotropic effects of metformin and exenatide.

Exenatide (a GLP-1 agonist) expresses anti-inflammatory properties in cultured human monocytes/macrophages in a protein kinase A and B/Akt manner.

BACKGROUND: Incretin-based therapies in the treatment of type 2 diabetes mellitus are associated with significant improvements in glycemic control, which are accompanied by a beneficial impact on atherosclerosis. Macrophages are essential in the development of atherosclerotic plaques and may develop features that accelerate atherosclerosis (classically activated macrophages) or protect arterial walls against it (alternatively activated macrophages). Therefore, we explored whether beneficial actions of exenatide are connected with the influence on the macrophages' phenotype and synthesis of inflammatory and anti-inflammatory cytokines. METHODS: Monocytes/macrophages were harvested from 10 healthy subjects. Cells were cultured in the presence of exenatide, exendin 9-39 (GLP-1 antagonist), LPS, IL-4, PKI (PKA inhibitor) and triciribine (PKB/Akt inhibitor). We measured the effects of the above-mentioned compounds on markers of macrophages' phenotype (inducible nitrous oxide (iNOS), arginase 1 (arg1) and mannose receptors) and concentration of nitrite, IL-1ß, TNF-α and IL-10. RESULTS: Exenatide significantly increased the level of IL-10 and decreased both TNF-α and IL-1ß in LPS-treated monocytes/macrophages. Furthermore exenatide increased the expression of arg1-a marker of classical activation and reduced the LPS-induced expression of iNOS-a marker of classical activation. According to experiments with protein kinases inhibitors we found that proinflammatory markers were protein kinase A dependent, whereas the activation of alternative activation was similarly reliant on protein kinase A and B/Akt. CONCLUSIONS: We showed that exenatide skewed the macrophages phenotype toward anti-inflammatory phenotype and this effect is predominantly attributable to protein kinase A and to a less extent to B/Akt activation.

[Novel strategies of ovarian cancer treatment]. / Nowe strategie leczenia raka jajnika.

Ovarian cancer is the most lethal gynecological malignancy. Due to scarce specific symptoms, women usually seek medical help once the disease is highly advanced, with distant metastases. Currently no screening is available, making this particular cancer hard to detect in the early stage. Standard treatment is insufficient for many patients, especially in the recurrent disease. This fact explains the tremendous need to search for novel therapeutic approaches. Inhibition of angiogenesis and destruction of cancer stem cells are attempts that affect the tumor microenvironment. There is a lot of potential in inhibiting poly(ADP-rybose)polymerase (PARP) or I class histone deacetylase. Drug repositioning may also be beneficial, as it gives old drugs new purposes. Metformin, a well-known antidiabetic agent, is an example of this phenomenon. Constant progress in medicine and science makes us hope for positive outcomes while treating this highly dangerous disease.

[Oncolytic viruses as a new way of treatment of neoplastic diseases]. / Wirusy onkolityczne najnowszym sposobem walki z chorobami nowotworowymi.

Despite the unceasing progression in chemotherapy, radiotherapy and surgery, neoplasms are still the second, after cardiovascular diseases, cause of death in the world. The creation of oncolytic viruses gives hope for increase of anticancer therapy effectiveness. Oncolytic viruses are the type of viruses that selectively infect and cause the lyse of tumor cells excluding normal cells. This mechanism allows to avoid the consequences of the possible replication of the virus, which having entered to the organism, replicates in organism's cells by using the DNA of host cells. The development of genetic engineering and molecular biology has enabled the creation of this kind of genetically modified viruses, which deprive them of their virulence. Currently, there are many clinical trials in progress including the use of oncolytic viruses in head and neck squamous cell carcinoma, thyroid cancer, colorectal cancer, liver cancer, melanoma and glioblastoma multiforme treatment. There are parallel studies in animals using the subsequent viruses. Oncolytic viruses treatment is generally well tolerated, without significant side effects. It is worth to point out that this method combined with chemotherapy and radiotherapy allows to reduce the use of therapeutic doses, which significantly reduces the toxicity of conventional treatment. Further clinical studies evaluating the efficacy and safety of oncolytic viruses will develop more effective and better tolerated therapeutic protocols in the future.

Exenatide (a GLP-1 agonist) improves the antioxidative potential of in vitro cultured human monocytes/macrophages.

Macrophages are dominant cells in the pathogenesis of atherosclerosis. They are also a major source of reactive oxygen species (ROS). Oxidative stress, which is particularly high in subjects with diabetes, is responsible for accelerated atherosclerosis. Novel antidiabetic drugs (e.g., glucagon-like peptide-1 (GLP-1) agonists) were shown to reduce ROS level. Therefore, we conceived a study to evaluate the influence of exenatide, a GLP-1 agonist, on redox status in human monocytes/macrophages cultured in vitro, which may explain the beneficial effects of incretin-based antidiabetic treatment. Human macrophages obtained from 10 healthy volunteers were in vitro subjected to the treatment with GLP-1 agonist (exenatide) in the presence of lipopolysaccharide (LPS), antagonist of GLP-1 receptors (exendin 9-39), or protein kinase A inhibitor (H89). Afterwards, reactive oxygen species, malondialdehyde level, NADPH oxidase, and antioxidative enzymes [superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), and catalase] expression was evaluated. Finally, we estimated the activity of the abovementioned enzymes in the presence of H89. According to our findings, exenatide reduced ROS and malondialdyhyde (MDA) level by decreasing the expression of ROS-generating NADPH oxidase and by increasing the expression and activities of SOD and GSH-Px. We also showed that this effect was significantly inhibited by exendin 9-39 (a GLP-1 antagonist) and blocked by H89. Exenatide improved the antioxidative potential and reduced oxidative stress in cultured human monocytes/macrophages, and this finding may be responsible for the pleiotropic effects of incretin-based therapies. This effect relied on the stimulation of GLP-1 receptor.

A Novel, Highly Selective RT-QPCR Method for Quantification of MSRV Using PNA Clamping Syncytin-1 (ERVWE1).

HERV-W is a multi-locus family of human endogenous retroviruses (HERVs) that has been found to play an important role in human physiology and pathology. Two particular members of HERV-W family are of special interests: ERVWE1 (coding syncytin-1, which is a glycoprotein essential in the formation of the placenta) and MSRV (multiple sclerosis-associated retrovirus that is thought to play a significant role in human pathology as a result of its increased expression in the brain tissue and blood cells derived from patients with multiple sclerosis (MS)). Both ERVWE1 and MSRV mRNA share high level of similarity and hence a method that allows to exclusively quantify the MSRV expression in clinical samples would be desirable. We developed a quantitative polymerase chain reaction (QPCR) technique for the detection and quantification of the multiple sclerosis-associated retrovirus. The assay utilises fluorescently labelled oligonucleotide probe, which is complementary to the conservative fragment of MSRV env gene and a peptide nucleic acid (PNA) probe, fully complementary to the ERVWE1 sequence fragment that efficiently blocks the polymerase action on ERVWE1 templates. The PNA molecule, if used parallel with hydrolysis probe in QPCR analysis, greatly facilitates the detection efficiency of MSRV even if ERVWE1 is present abundantly in respect to MSRV in the analysed sample. We achieved a wide and measurable range from 1 × 10 e(2) to 1 × 10 e(8) copies/reaction; the linearity of the technique was maintained even at the low MSRV level of 1% in respect to ERVWE1. Using our newly developed method we confirmed that the expression of MSRV takes place in normal human astrocytes and in human umbilical vein endothelial cells in vitro. We also found that the stimulation of human monocytes did not influence the specific expression of MSRV but it caused changes in mRNA level of distinct HERV-W templates.

[Antiangiogenic therapy of malignant pheochromocytoma and paraganglioma with the view to the recent scientific developments]. / Antyangiogenna terapia zLoSliwej postaci pheochromocytoma i paraganglioma w Swietle najnowszych doniesieN naukowych.

Pheochromocytoma is a very rare tumor that stems from chromaffin cells and usually develops in the adrenal glands. Its equivalent, which exists outside of the adrenal glands, is paraganglioma. Approximately 10-26% of pheochromocytoma is malignant, what poses a significant therapeutical problem, as its presence, together with an abundant production of catecholamines, may lead to a number of perilous complications, such as spinal cord oppression or the damage of organs, what is responsible for producing catecholamines. Due to the risk that the tumor is, it is essential to event new and effective ways of treatment. In case of malignant tumors stemming from chromaffin cells, much is expected from antiangiogenic medicine. Its functioning consists of stopping of the process of neovascularization, which is indispensable for the development of the tumor. Sunitinib - a tyrosine kinase inhibitor - is perhaps the most promising antiangiogenic medicine, whose effectiveness is currently being evaluated in 2nd phase clinical trials. Attempts are also being made to conduct treatment with the use of other medicine of similar functioning, such as: thalidomide, imatinib or evrolimus.

[Drugs malabsorption after bariatric surgery]. / Zaburzenia wchlaniania leków u chorych po operacjach bariatrycznych.

Along with the increasing prevalence of obesity surgical methods of treatment are becoming more accessible. The success of bariatric surgery results from the failure of conservative treatment of obesity through diet, exercises and behavioral changes. Such behaviors, including less effective drug therapy, are responsible for rather small and unstable weight reduction. In contrast, surgical treatment restrictions limiting the amount of food intake, excluding treatments and mixed treatments usually give a spectacular effect, though it is not always consistent with the previous assumptions. Most of us, however, do not realize the risk of the consequences that these treatments bring. The change of the absorption of oral medications, vitamins, microelements, often change the way they are metabolized and it forces ones to change the form of the drug, the way they are taken, which undoubtedly is associated with higher costs of later treatment. Many clinicians are unaware of the need for lifelong supplementation of some vitamins and microelements, as well as the need to modify pharmacotherapy in patients who have undergone this type of operations. This article presents examples of the best known and described changes in the absorption of drugs and micronutrients and explains how well oriented the internist ought to be when qualifying a patient to undergo this type of treatment. The described topic is interesting, but also controversial due to the fact that it corresponds closely with the surgical specialties, quite spectacular in their effects and behavioral specializations, that in many years time will be looking after the patient who underwent the surgery, but is still sick.

[The newest perspectives on the treatment of glioblastoma multiforme]. / Najnowsze perspektywy w leczeniu glejaka wielopostaciowego.

Glioblastoma multiforme is the most common and most malignant primary brain tumor, with a yearly incidence of 2.5 in 100,000. It has a very dismal prognosis, since the medium overall survival of untreated patients is as low as 3 months. Location in the central nervous system, high aggressiveness, spreading alongside blood vessels and white matter, cause a lot of therapeutic challenges. The blood-brain barrier unables most of the systemic drugs to reach the tumor and complete resection is usually impossible. Because of that, effects of the standard treatment remain unsatisfying. It forces to search for novel treatment options. Regarding pharmacotherapy a lot of attention is brought to antiangiogenic therapies, where the most common drug is bevacizumab. In Europe it is registered to use in diffuse breast cancer, non-small cell lung cancer, colon and rectal cancer with metastases, but for glioblastoma it's use is still considered to be experimental. Inhibition of integrins, extracellular matrix metalloproteinases and EGFR are among other therapeutic goals. There is a broad range of studies on breaking the resistance of cancer stem cells, modifying the niche of cancer cells, active immunotherapy and the use of microRNAs. The field of stereotactic radiosurgery is also under constant improvement. Methods of both genetic and biomedical engineering, such as nanotubes or liposomes, can be helpful to overcome the blood-brain barrier and insert the drugs directly and even selectively into the tumor.

[Hyperthermic intraperitoneal chemotherapy as a new way of peritoneal metastases treatment]. / Dootrzewnowa chemioterapia perfuzyjna w hipertermii jako nowy sposób leczenia przerzutów nowotworowych do otrzewnej.

Peritoneal neoplastic disease may originate primarily from neoplasms developing in peritoneum or metastases of cancers mostly arising in digestive or female reproductive systems. Peritoneal neoplasia leads to a much poorer prognosis and in most cases the palliative care, which focus on gastrointestinal motility and patency, adequate analgesia and psychological care, is being introduced. Systemic chemotherapy is ineffective as a result of existence of physiological blood-peritoneal barrier that blocks hydrophilic chemotherapeutics. Nowadays there are attempts to eliminate this problem by performing cytoreductive surgery accompanied by hyperthermic intraperitoneal chemotherapy (HIPEC). Undoubtedly the possibility to gain high local concentration of chemotherapeutic directly in peritoneal cavity is primary advantage of this method. In the carried out clinical trials the efficiency of this procedure in prolonging patient life in a relatively good comfort was proven. This method is highly recommended in case of peritoneal metastases from colorectal cancer treatment. There are ongoing studies focused on proving the effectiveness of HIPEC in case of other types of neoplasms and use of that therapy as a prophylactic treatment before metastases occur.

Comparison of chosen activation markers of human monocytes/macrophages isolated from the peripheral blood of young and elderly volunteers.

BACKGROUND: The immune system of humans is strongly affected by the processes of aging and what is called immunosenescence and inflammaging. Aging processes are also associated with altered macrophage functions and their ability to undergo differential activation. As a result, the risk of macrophage-related disorders like atherosclerosis is increased in the elderly. METHODS: Human monocyte-derived macrophages obtained from young and elderly healthy volunteers were stimulated with either lipopolysaccharide (LPS) or interleukin-4 (IL-4), and the expression of classical and alternative activation markers was assessed. The concentrations of nitric oxide (NO), reactive oxygen species (ROS) and IL-1ß were measured in addition to the expression of genes and relevant proteins of inducible nitric oxide synthase, IL-1ß, arginase-1 and suppressor of cytokine signaling-1. RESULTS: We showed that the macrophages isolated from the young generally demonstrated higher responsiveness to introduced stimuli and balanced the classical activation state. The cells from the elderly showed stronger generation of nitric oxide (NO) and reactive oxygen species (ROS), which contribute to stress and damage reactions. CONCLUSIONS: The changes observed in the macrophages isolated from the elderly indicate that these cells could contribute to the development of metabolic disorders like atherosclerosis and diabetes. The cells from the young volunteers are less likely to present such properties.

Metformin affects macrophages' phenotype and improves the activity of glutathione peroxidase, superoxide dismutase, catalase and decreases malondialdehyde concentration in a partially AMPK-independent manner in LPS-stimulated human monocytes/macrophages.

BACKGROUND: Diabetic patients experience accelerated atherosclerosis. Metformin is a cornerstone of the current therapy of type 2 diabetes. Macrophages are the key cells associated with the development of atherosclerotic plaques. Therefore, our aim was to assess the in vitro effects of metformin on macrophages and its influence on the mechanisms involved in the development of atherosclerosis. MATERIALS AND METHODS: Peripheral blood mononuclear cells were obtained from the group including 16 age-matched healthy non-smoking volunteers aged 18-40 years. Monocytes were further incubated with metformin, LPS and compound C--a pharmacological inhibitor of AMPK. The impact of metformin on oxidative stress markers, antioxidative properties, inflammatory cytokines and phenotypical markers of macrophages was studied. RESULTS: We showed that macrophages treated with metformin expressed less reactive oxygen species (ROS), which resulted from increased antioxidative potential. Furthermore, a reduction in inflammatory cytokines was observed. We also observed a phenotypic shift toward the alternative activation of macrophages that was induced by metformin. All the aforementioned results resulted from AMPK activation, but a residual activity of metformin after AMPK blockade was still noticeable even after inhibition of AMPK by compound C. CONCLUSIONS: Authors believe that metformin-based therapy, a cornerstone in diabetes therapy, not only improves the prognosis of diabetics by reducing blood glucose but also by reducing oxidative stress, inflammatory cytokine production and the shift toward alternative activation of macrophages.

A peptide nucleic acid (PNA)-mediated polymerase chain reaction clamping allows the selective inhibition of the ERVWE1 gene amplification.

A new peptide nucleic acid (PNA) mediated QPCR technique for the detection and quantification of the Multiple Sclerosis-Associated Retrovirus (MSRV) belonging to the human endogenous retrovirus-W (HERV-W) family has been developed. The assay utilizes a PNA probe which is fully complementary to the ERVWE1 sequence, another member of the HERV-W family which is closely related to MSRV. Due to its excellent affinity to a completely matched template, PNA probe selectively blocks the amplification of ERVWE1 thus allowing the specific and exclusive detection and quantification of the MSRV as PNA does not interfere with the amplification of MSRV. Using this newly developed method we found that MSRV is predominantly expressed over ERWVE1 in astrocyte-derived U-87 MG cell line but not in human umbilical vein endothelial cells or human placental cells.

The influence of ezetimibe on classical and alternative activation pathways of monocytes/macrophages isolated from patients with hypercholesterolemia.

Macrophages are crucial for the development of atherosclerotic plaques. Classically activated macrophages contribute to plaque growth and destabilization, while alternatively activated macrophages increase plaque stability. Here, we assessed the influence of ezetimibe on the activation of monocyte-derived macrophages isolated from patients with hypercholesterolemia (total cholesterol 263.4 ± 12.5 mg/dl, low-density lipoprotein cholesterol 179.7 ± 11.3 mg/dl, triglycerides 123.9 ± 11.4 mg/dl). Cells were stimulated with 1 µg/ml lipopolysaccharide (LPS) or 1 µg/ml LPS plus 22 ng/ml ezetimibe. Control cells were left unstimulated. The expression of classical activation markers (interleukin-1ß (IL-1ß), nitric oxide (NO), and inducible nitric oxide synthase (iNOS)) and alternative activation markers (mannose receptor (MR) and arginase-1 (Arg1)) was determined after 48 h. The employed analytical methods included enzyme-linked immunosorbent assay, Griess reaction, real-time polymerase chain reaction, and Western blotting. LPS increased the secretion of IL-1ß and NO and the expression of iNOS mRNA, iNOS protein, and Arg1 protein. It did not affect the expression of MR or Arg1 mRNA. In comparison to LPS stimulation, co-stimulation with ezetimibe decreased the secretion of IL-1ß and the expression of iNOS mRNA and protein, while it increased MR mRNA and protein expression. Co-stimulation with ezetimibe did not change the secretion of NO or the expression of Arg1. The results suggest that ezetimibe in inflammatory in vitro conditions contributes to the suppression of classical and promotion of the alternative macrophage activation.

[Metformin as a key to alternative activation of microglia?]. / Metformina kluczem do alternatywnej aktywacji mikrogleju?

The results of recent studies suggest that metformin, in addition to its antihyperglycemic efficacy, may also attenuate neuroinflammation and directly act on the central nervous system. However, the molecular mechanisms by which metformin exerts its anti-inflammatory effects in the brain remain largely unknown. Adenosine-monophosphate-activated protein kinase (AMPK) activation is the most well-known mechanism of metformin action. However, some of the biological responses to metformin (e.g. the release of cytokines and the expression of arginase I or PGC-1α) are not limited to AMPK activation but also are mediated by AMPK-independent mechanisms. This article reviews current evidence supporting the hypothesis that the shift of microglia toward alternative activation may underlie the beneficial effects of metformin observed in animal models of neurological disorders.

Eplerenone mimics features of the alternative activation in macrophages obtained from patients with heart failure and healthy volunteers.

Alternative activation of macrophages plays protective role in cardiac remodelling in heart failure and the activity of mineralocorticoid receptor may determine the phenotype of these cells. We examined the influence of eplerenone, aldosterone, and IL-4 on descriptors of alternative activation in blood monocytes collected from 19 patients with heart-failure and 20 healthy volunteers. "Heart failure" macrophages in comparison with "healthy" macrophages had increased mineralocorticoid activity, NO and reactive oxygen species production, expression of iNOS mRNA and protein, but decreased expression of arginase I and mannose receptor proteins, and activity of MnSOD and CuZnSOD. Aldosterone increased mineralocorticoid activity, NO and reactive oxygen species production, iNOS mRNA and protein expression, MnSOD and CuZnSOD activity. Eplerenone attenuated the effects of aldosterone on all but MnSOD and CuZnSOD variables. Eplerenone alone increased the production of NO, MnSOD and CuZnSOD activity, arginase I gene and protein expression, and mannose receptor gene and protein expression, but decreased mineralocorticoid activity only in "heart failure" macrophages. The latter suggests altered function of mineralocorticoid receptor in heart failure. Increased mineralocorticoid activity accounts for increased NO production, iNOS gene and protein expression but does not explain the increased basal reactive oxygen species production and decreased markers of alternative activation in "heart failure" macrophages. In the lack of change in basal mineralocorticoid activity, eplerenone increases markers of alternative activation in a mineralocorticoid receptor-independent manner. Because of changes in iNOS and NO variable, eplerenone induced qualitatively different activation of macrophages from that obtained with IL-4.