Investigation of the effects of urotensin II receptors in LPS-induced inflammatory response in HUVEC cell line through calcineurin/NFATc/IL-2 pathway.

PURPOSE: The effect of urotensin II (U-II), a powerful endogenous vasoconstrictor substance, on the immune system and its mediators is very important. It was herein aimed to demonstrate the possible relationship between the calcineurin/nuclear factor of activated T-cells cytoplasmic 1/interleukin-2 (CaN/NFATc/IL-2) pathway and urotensin receptors (UTRs) in inflammatory response due to lipopolysaccharide (LPS). METHODS: An LPS-induced inflammation model was used on the human umbilical vein endothelial cells (HUVEC) cell line and drugs were applied accordingly, forming the following groups: Control Group, LPS Group, Agonist Group (10-8 â€‹M U-II), Antagonist Group (10-6 â€‹M palosuran), Tacrolimus (TAC) Group (10 â€‹ng/mL FK-506), Agonist â€‹+ â€‹TAC Group, and Antagonist â€‹+ â€‹TAC Group. Gene expression analyses were performed using real-time polymerase chain reaction (RT-PCR). RESULTS: In the analysis of the cell viability at 48 and 72 â€‹h, there was a decrease in the Agonist Group, while in the Agonist â€‹+ â€‹TAC Group, the cell viability increased. In the Antagonist Group, cell viability was maintained when compared to the LPS Group, while in the TAC Group, this effect was reduced. The mRNA expression levels of UTR, CaN, NFATc, IL-2 receptor (IL-2R), IL-6 and nuclear factor kappa B (NF-κB) were higher in the LPS Group than in the Control Group, and even the UTR, CaN, NFATc, IL-2R were higher with agonist administration. This effect of the agonist was shown to be completely mitigated in the presence of the CaN inhibitor. CONCLUSION: U-II and its receptors can perform key functions regarding the endothelial cell damage via the CaN/NFATc/IL-2 pathway.

Relationship Between Angiotensin II Type 1 Receptor Antibody Positivity and Cytokine Gene Polymorphism in Renal Transplant Patients When Organ Rejection Occurs.

OBJECTIVES: Kidney transplant remains the gold standard for the treatment of end-stage renal disease. Relationships between the presence of non-HLA antibodies, antibodies to AT1R, and cytokine gene polymorphisms with rejection have recently been shown. We sought to determine whether the presence of antibodies to AT1R and cytokine gene polymorphisms affected the development of rejection in pediatric and adult patients, whether a relationship is present between cytokine polymorphism and level of antibodies to AT1R, and whether their presence can be a biomarker pretransplant. MATERIALS AND METHODS: Our study included 100 pediatric and adult kidney transplant patients plus 50 healthy controls. Levels of AT1R antibodies (by enzyme-linked immunosorbent assay) and gene polymorphisms of the cytokines transforming growth factor ß, tumor necrosis factor α, interleukins 6 and 10, and interferon gamma cytokines (by sequence- specific primer-polymerase chain reaction) were studied retrospectively and evaluated with the SPSS statistical program. RESULTS: We found no statistically significant relationship between levels of antibodies to AT1R and gene polymorphisms among the studied cytokines in patients with rejection compared with the healthy controls and patients with uneventful courses posttransplant. However, higher levels of antibodies to AT1R were observed in pediatric compared with adult transplant recipients (P < .001). A statistically significant relationship was also observed between transforming growth factor ß1 C/C G/C low-release and interleukin 6 G/C high-release gene polymorphism and levels of antibodies to AT1R (P < .001). CONCLUSIONS: Because we observed that some gene polymorphisms among the studied cytokines may affect AT1R antibody levels, future studies are needed to understand the mechanism of the relationship. In addition, studies with larger groups are required to sufficiently confirm that higher antibody levels are present in pediatric versus adult patients.

Caspase-1 and interleukin-18 in children with post infectious bronchiolitis obliterans: a case-control study.

The exact immunological mechanisms of post infectious bronchiolitis obliterans (PIBO) in childhood are not fully known. It has been shown that the inflammasome and IL-18 pathway play important roles in the pathogenesis of lung fibrosis. We aimed to investigate the role of caspase-1, IL-18, and IL-18 components in PIBO. From January to May 2020, children with PIBO, children with history of influenza infection without PIBO, and healthy children were asked to participate in the study in three pediatric pulmonology centers. Serum caspase-1, IL-18, IL-18BP, IL-18R, and INF-γ levels were measured by ELISA and compared between the 3 groups. There were 21 children in the PIBO group, 16 children in the influenza group, and 39 children in the healthy control group. No differences in terms of age and gender between the 3 groups were found. IL-18 and IL-18BP levels were higher in the healthy control group (p = 0.018, p = 0.005, respectively). IL-18R was higher in the PIBO group (p = 0.001) and caspase-1 was higher in the PIBO and influenza group than the healthy control group (p = 0.002). IFN-γ levels did not differ between the 3 groups. IL-18BP/IL-18 was higher in the influenza group than the PIBO group and the healthy control group (p = 0.003). CONCLUSIONS: Caspase-1 level was increased in patients with PIBO which suggests that inflammasome activation may have a role in fibrosis; however, IL-18 level was found to be low. Mediators other than IL-18 may be involved in the inflammatory pathway in PIBO. Further immunological studies investigating inflammasome pathway are needed for PIBO with chronic inflammation. WHAT IS KNOWN: • Post infectious bronchiolitis obliterans (PIBO) is a rare, severe chronic lung disease during childhood which is associated with inflammation and fibrosis which lead to partial or complete luminal obstruction especially in small airways. • The exact immunological mechanisms of PIBO in childhood are not fully known. WHAT IS NEW: • Inflammasome activation persists even years after acute infection and may play a role in fibrosis in PIBO. • Mediators other than IL-18 may be involved in these inflammatory pathway.

Evaluation of systemic inflammatory response in cardiovascular surgery via interleukin-6, interleukin-8, and neopterin.

AIM: The aim of this study was to evaluate the serum levels of interleukin-6 (IL-6), IL-8, and neopterin as a sign of systemic inflammatory response syndrome after open-heart surgery. In this study, we evaluated the influences on the levels of IL-6, IL-8, and neopterin of coronary artery bypass grafting (CABG) and valve replacement surgeries with and without the use of extracorporeal circulation (ECC). MATERIALS AND METHODS: This prospective study was performed in 30 patients. In this study, we evaluated patients who underwent valve replacement surgery (group 1, n = 10), CABG with ECC (group 2, n = 10), or CABG using the beating-heart technique (group 3, n = 10). With the Human Investigation Ethics Committee consent, blood samples were obtained from the patients before the surgery (T0) and after 1 hour (T1), 4 hours (T2), 24 hours (T3), and 48 hours (T4) of protamine injection. IL-6, IL-8, and neopterin levels were measured using commercial enzyme-linked immunosorbent assay kits. RESULTS: The demographic data and preoperative and operative characteristics of the patients were similar. Neopterin IL-6 and IL-8 levels significantly increased first at the fourth hour after the surgery. When compared to the levels before the surgery, this increase was statistically significant. Unlike the other 2 groups of patients, those who experienced CABG with the beating-heart technique (group 3) had decreased neopterin levels at the first hour after the surgery, but this decrease was not statistically significant. Neopterin levels increased later in the OPCAB group, but these increased levels were not as high as the neopterin levels of groups 1 and 2. Neopterin reached maximum levels at the 24th hour and, unlike groups 1 and 2, in group started to decrease at the 48th. CONCLUSIONS: Complement activation, cytokine production, and related cellular responses are important factors during open-heart surgery. It is certain that ECC activates the complement systems, and activated complement proteins cause the production of several cytokines. In our study, neopterin levels in patients who underwent beating-heart method surgery were lower than those in the other groups, and these levels started to decrease at the 48th hour. These data suggest that the systemic inflammatory response was less activated in that patient group. The beating-heart method might be an important alternative in CABG surgery to minimize the complications and mortality related to surgery.

Effects of different medical treatments on serum copper, selenium and zinc levels in patients with rheumatoid arthritis.

The aim of the present study was to measure the changes in serum selenium, zinc, and copper in patients being treated for rheumatoid arthritis. Thirty-two patients and 52 healthy controls were included in the study. The copper level was higher and those of selenium and zinc were lower in the patients relative to controls. Treatment with methotrexate elevated the zinc levels, but not zinc and selenium. Treatments with salazopyrin, corticosteroids, chloroquine, and non-steroidal anti-inflammatory drugs did not change the levels of any of the elements studied. The decrease in zinc and selenium levels and elevation in copper levels observed in the patients probably resulted from the defense response of organism and are mediated by inflammatory-like substances.

Neopterin levels and dexamethasone suppression test in obsessive-compulsive disorder.

Neopterin, a biopterin precursor that is released by macrophages, is an important immunological marker in psychiatric disorders. It has been reported that glucocorticoids may cause suppression of cell-mediated immunity and consequently result in decreased neopterin levels. In the present study, we evaluated whether dexamethasone suppression test (DST) and neopterin findings were associated with pure obsessive-compulsive disorder (OCD) patients (OCD-D group) and the concomitant OCD and depression (OCD+D group). The sample comprised 44 patients with OCD (27 with OCD-D and 17 with OCD+D) and 30 control subjects. There was significantly higher DST nonsuppression in the OCD+D group than in the OCD-D group. With regard to mean neopterin levels, there was no significant difference between the OCD-D group and the control group, but there was a statistically significant difference between the OCD+D group and the control group. The OCD+D group had significantly lower neopterin levels than the 20 OCD-D group. We suggest that this distinction may reflect the fact that glucocorticoids can lead to suppression of cell-mediated immunity and consequently can result in decreased neopterin levels. In conclusion, our results suggest that not the OCD-D group had normal neopterin levels and DST results, and also that OCD may be a heterogeneous subtype characterized by some biological indicators or anxiety and affective disorders.

Effect of chronic exercise on immunoglobin, complement and leukocyte types in volleyball players and athletes.

BACKGROUND: The objective of this study was to investigate the effect of various sports disciplines on basic elements of the acquired and natural defense systems and to compare their results with that of non-sporting controls. MATERIALS/METHODS: The study included 15 sedentary (Group 1), 15 volleyball players (Group 2) and 15 long distance running athletes (Group 3). The VO2 Max for the groups was determined by subjecting the individuals to a 20 meter Shuttle run test. Immunoglobin and complement levels were measured by the turbidometric method. Leukocyte counts were made through peripheral blood smear examination. RESULTS: The IgA levels in sedentary living individuals was found to be lower than that in volleyball and athlete groups while that in athletes was in turn lower than that in volleyball players (p<0.05). In volleyball players the IgG level was significantly higher than both the sedentary living individuals and the athletes (p<0.05). IgM however, showed a significantly higher level in the sedentary group than in both athletes and volleyball players. The C3 levels in athletes was significantly lower than that of volleyball players and sedentary individuals (p<0.05), while the C4 levels were higher in the volleyball group (p<0.05). The leukocyte, eosinophil, and monocyte counts in the athletes were also significantly lower than those in the volleyball and sedentary groups (p<0.05). The values for the sedentary group on the other hand however, was found to be lower than those of the volleyball players (p<0.05). The neutrophil counts was lower in the sedentary group than in the volleyball group (p<0.05). The levels in the athlete group was lower than that in the volleyball group, however (p<0.05). The lymphocyte counts in volleyball players was higher than that in the sedentary group (p<0.05). CONCLUSIONS: In conclusion, changes in the immune parameters were found to be dependent on the type of physical exercise performed on regular basis. Considering the protective antibody response in those performing regular sports, moderate exercise carried out on regular basis can be said to affect positively mononuclear and polymorphonuclear phagocytic cells--the basic elements of the natural immune system.

The effects of exercise on the immune system and stress hormones in sportswomen.

BACKGROUND: Despite the numerous studies controversial results exist in specific immune response to exercise. The aim of this study was to determining the differences in the humoral immune parameters, serum ACTH and cortisol levels existing between sportswomen and sedentary subjects and the effect of acute aerobic and anaerobic exercise on these parameters. MATERIALS/METHODS: 40 sportswomen (Groups 1 and 2) and 20 sedentary women (Group 3) were enrolled and Group 1 performed aerobic exercise on a treadmill for 30 minutes while the Group 2 was subjected to the Wingate effort test for 30 seconds. Before exercising (at 8.30 a.m), immediately after the exercise, and 4 hours, days 2 and 5 days after exercising blood samples were obtained and the levels of IgA, IgG, IgM, complement 3 (C3) and complement 4 (C4) were determined turbidometrically. RESULTS: Before exercise: the means of IgA and IgG values in the G1 and G2 groups were higher than the Group G3, and the mean cortisol levels in the sedentary group was significantly higher (p<0.05). After Exercise: Whereas the C3 and C4 levels decreased significantly at the end of the exercise in Group 1 and 2 (p<0.05), the IgA, IgG (p<0.05) and IgM levels (p<0.01) at the 4th and 5th determinations were observed to be significantly higher in only the Group 1. The cortisol and ACTH levels were found to have increased significantly (p<0.05) in the Group 1. In Group 2, no changes were observed in the IgA, IgG and IgM levels. CONCLUSIONS: We conclude that regular and moderate exercise has favorable effects on the immune system by increasing immunoglobulins which are potent protective factors.

Procalcitonin, c-reactive protein and neopterin levels in patients with coronary atherosclerosis.

OBJECTIVE: Recent studies demonstrate that the serum inflammatory markers increase in patients with atherosclerosis. We aimed to assess whether a difference exists between patients with acute coronary syndrome and patients with stable angina pectoris in respect to serum neopterin, procalcitonin (PCT) and C-reactive protein (CRP) levels. METHODS AND RESULTS: A total of 52 patients (42 male, 10 female) who had atherosclerosis confirmed by angiography and were being followed up for an acute coronary syndrome were recruited and for control group, 45 patients with stable angina pectoris (SA) (35 male and 10 female) who underwent coronary angiography, were examined. Serum concentrations of neopterin, CRP and PCT in the study group (acute coronary syndrome) were compared to control group (stable angina pectoris). The mean neopterin level of the study group was 22.47 +/- 2.93 nmol/l, the mean CRP level was 30.40 +/- 8.05 mg/l and the mean PCT level was 0.40 +/- 0.04 ng/ml. In control group these levels were 12.26 +/- 0.61 nmol/l (p < 0.05), 5.26 +/- 0.64 mg/l (p < 0.001) and 0.19 +/- 0.02 ng/ml (p < 0.001), respectively. CONCLUSION: In the presented study our results showed that these markers can be useful for the assessment of inflammation related to atherosclerosis.

Determination of intracellular cytokines produced by Th1 and Th2 cells using flow cytometry in patients with brucellosis.

The aim of the study was to evaluate intracellular interferon-gamma (IFN-gamma), and interleukin-4 (IL-4) levels in pre- and post-treatment periods of brucellosis patients and to determine the relationship between these parameters and patients' clinical findings. Twenty-five patients diagnosed as brucellosis and 11 aged-matched healthy volunteers were included in the study. CD3+CD4+ T lymphocytes levels were significantly lower in patients with brucellosis as compared to the control group. CD3+CD8+ T lymphocytes and CD3+IFN-gamma+ levels were increased in brucellosis patients compared with the control group. CD4+IFN-gamma+ and CD4+IL-4+ levels were no different between patients and healthy individuals. CD3+IL-4+ levels decreased in patients compared with healthy controls. Pre-treatment CD3+IFN-gamma+ levels dramatically increased in patients responsive to management compared with the unresponsive ones. In responsive cases, CD3+IFN-gamma+ levels decreased statistically after the treatment while in unresponsive cases no meaningful change was observed with respect to treatment. Adding IFN-gamma to the treatment for improving the depleted levels of IFN-gamma can be beneficial in patients with brucellosis who shows a tendency to chronicity or patients who do not respond to the treatment.

The role of allergy in the etiology of otitis media with effusion; immune system and cytokines.

OBJECTIVES: Otitis media with effusion (OME) is one of the major causes of hearing loss in childhood. In this study, the role and the importance of allergy in the etiology of OME was evaluated with reference to the immune system and the cytokines. PATIENTS AND METHODS: Seventy-eight ears of 59 patients who had undergone myringotomy with the diagnosis of OME were included in this study. Serum samples of 26 healthy children who were in the same age group and were living in the same region under similar socioeconomic circumstances were examined as the control group. RESULTS: Interferon (IFN)-gamma levels were significantly lower and IL-4 and IgE levels were significantly higher in the serum samples of the patient group when compared with those of the control group (p<0.05). A positive correlation was found between IL-4 and IgE in the serum samples of patients (p<0.01). However, there were negative correlations between IFN-gamma and both IgE and IL-4 levels, that were not statistically significant. CONCLUSION: T-helper polarization suggests that allergy may have a role in the etiology of OME. Therefore, it should be kept in mind in the treatment of OME.

Antimonial therapy induces circulating proinflammatory cytokines in patients with cutaneous leishmaniasis.

The objective of this study was to evaluate the association between antimonial therapy and circulating levels of proinflammatory cytokines in patients with cutaneous leishmaniasis (CL). Patients were treated with conventional chemotherapy by using pentavalent antimonium salts (Glucantime) for 3 weeks. Circulating plasma levels of the proinflammatory cytokines interleukin-1beta (IL-1beta), IL-6, IL-8, and tumor necrosis factor alpha (TNF-alpha) were determined for CL patients and healthy subjects before and 3 weeks after the treatment was started. Plasma IL-1beta, IL-6, IL-8, and TNF-alpha levels were significantly higher for pretreatment CL patients than for healthy subjects. Proinflammatory cytokines significantly increased after 21 days postinfection compared to levels for the pretreatment patients. These increments were approximately 3-fold for IL-1beta and TNF-alpha levels, 10-fold for IL-6 levels, and 20-fold for IL-8 levels in patients with CL. Taken together these results indicate that circulating proinflammatory cytokine levels were increased in patients with CL as a consequence of host defense strategies, and antimonial therapy may induce these cytokines by affecting the macrophage or other components of the host defense system.

Trace elements in viral hepatitis.

In this study, serum trace elements, including selenium (Se), zinc (Zn), copper (Cu), were determined by using Atomic Absorption Spectrophotometer (SpectrAA 250 Plus Zeeman, Varian, Australia) in sera of patients with viral hepatitis (A, B, C, D, E) cases (n = 102), and statistically compared with the controls (n = 52). In viral hepatitis, Cu levels were found as 3.23 +/- 1.02 mg/L, and this value was significantly higher than the control group (1.13 +/- 0.21) (p < 0.01). Both, Se and Zn levels found to be significantly low in viral hepatitis cases (p < 0.01). While Se level was 81.4 +/- 26.01 microg/L in viral hepatitis (n = 101), it was found to be 166.15 +/- 4.58 microg/L in healthy individuals. Meanwhile, Zn levels were 0.230 +/- 0.081 mg/L and 0.748 +/- 0.392 mg/L in hepatitis cases (n = 101) and the control group, respectively. There was no difference amongst viral hepatitis groups classified in regard with agents and clinical manifestation, such as A, acute hepatitis B, chronic hepatitis B, C, D and E. Previously, it was indicated that absorption disorders in gastrointestinal system, especially in chronic cases, were not main causes of decrease of trace elements by iron and several other parameters in sera of the cases. Therefore, we suggest that decrease in Zn and Se levels and elevation in Cu levels are probably resulted from defence strategies of organism and induced by the hormone-like substances.

Immunological follow-up of hydatid cyst cases

Hydatid disease is caused by Echinococcus granulosus. In this study, we aimed to investigate the benefit of monitoring cases with hydatid cyst by means of immune components in patients in a long-term follow-up after surgery. Eighty-four preoperative and postoperative serum samples from 14 cases undergoing surgery for hydatid disease were evaluated in terms of immune parameters, such as total and specific IgE, IgG, IgM, IgA and complement. Total and specific IgE were determined by ELISA. Specific IgG levels were measured by indirect hemaglutination.Total IgG, IgM, IgA and complement (C3 and C4) were detected by nephelometry. Imaging studies were also carried out during the follow-up. In none of the patients hydatid cysts were detected during the follow-up. Total IgE levels in the sera of the patients decreased to normal six months after surgery. Although specific IgE against echinococcal antigens decreased one year after operation, levels were still significantly high. There were no changes in the levels of anti-Echinococcus IgG and total IgG in follow-up period. Additionally, other parameters, such as IgA, IgM, C3 and C4, were not affected

Adenosine deaminase activities in sera, lymphocytes and granulocytes in patients with cutaneous leishmaniasis

Adenosine deaminase (ADA) activities in sera, lymphocytes and granulocytes in patients with cutaneous leishmaniasis were investigated and compared with control groups. Fifty patients and 50 healthy individuals were studied. The clinical diagnosis was parasitologically confirmed by culture and Giemsa stain. ADA activities were measured by colorimetric method. Serum ADA activities 37.80 ñ 11.90, 18.28 ñ 6.08 IU/L (p<0.0001), lymphocyte specific ADA activities 14.90 ñ 7.42, 8.38 ñ 7.42 U/mg protein (p=0.04), granulocyte specific ADA activities 1.15 ñ 0.73, 1.09 ñ 0.67 U/mg protein (p>0.05) were found in patients and control groups, respectively. ADA activity increases in some infectious diseases were cell mediated immune mechanisms are dominant. In cutaneous leishmaniasis, lymphokine-mediated macrophage activity is the main effector mechanism. Increase in serum and lymphocyte ADA activities in patients with cutaneous leishmaniasis may be dependent on and reflects the increase in phagocytic activity of macrophages and maturation of T-lymphocytes.