TNF-alpha Concentrations Research Articles

A Rediscovered Immunomodulatory Action of Cancer Chemotherapy to Pilot the Cytokine Network in an Antitumor Way

The recent proposal of cancer immunotherapy with anti-checkpoint inhibitor monoclonal antibodies, as well as the previous immunotherapy with IL-2, would require an interpretation of cancer chemotherapy not only in terms of therapeutic strategy carried out to destroy cancer cells, but also as an approach potentially able to influence and modulate the cytokine network in an attempt to correct cancer-related cytokine alterations responsible for tumor progression itself. This statement is justified by the fact that the efficacy of cancer immunotherapy is depending at least in part on the cytokine secretions of patients. Despite the great number of cancer-related cytokine alterations, the main alterations provided by a physiopathological and prognostic significance would consisting of low blood levels of IL-2 and IL-12 in association with abnormally high concentrations of IL-1, IL-6, TNF-alpha, IL-10 and TGF-beta. The effects of chemotherapy on cytokine secretions have appeared to depend on the type of agent, as well as on its dosage and schedule of administrarion. At present, the main effects of chemotherapy provided by a potential clinical application would be represented by the stimulatory action of adriamycin on IL-2 secretion, the inhibitory effect of cisplatin on IL-6 secretion, the stimulatory effect of gemcitabine on IL-12 production, and the inhibitory action of both cyclophosphamide and gemcitabine on regulatory T cell system. Then, the future chemo-immunotherapeuticregimenswouldhavenot to be elaborated not only on the bases of empiristic criteria and on the cytotoxic effects of che various chemotherapeutic agents, but also by taking into consideration their specific activity on the secretion of those cytokines, whose anomalous production has to be corrected and neutralized.

Effects of Betulinic Acid Derivative on Lung Inflammation in a Mouse Model of Chronic Obstructive Pulmonary Disease Induced by Particulate Matter 2.5.

BackgroundChronic obstructive pulmonary disease (COPD) is mainly induced by the increased content of particulate matter 2.5 (PM2.5) in the atmosphere. This study aimed to evaluate the effects of betulinic acid derivative on lung inflammation in a mouse model of chronic obstructive pulmonary disease induced by particulate matter 2.5.Material/MethodsThe mice were given a PM2.5 (25 μl) suspension for 7 days by the intranasal route to establish a COPD model. The content of TNF-α and IL-6 in the BALF samples was measured by commercially available ELISA kits.ResultsThe PM2.5-induced higher LDH and ACP levels were significantly alleviated in mouse lung tissues by treatment with betulinic acid derivative. Treatment with betulinic acid derivative also suppressed PM2.5-induced increase in AKP and ALB levels in mouse lung tissues. Betulinic acid derivative reversed PM2.5-mediated suppression of SOD activity and elevation of NOS level in mouse BALF. Moreover, the PM2.5-induced excessive NO and MDA levels in mouse BALF were significantly reduced (P<0.05) by treatment with betulinic acid derivative. Treatment with betulinic acid derivative improved TNF-α and IL-6 levels in BALF induced by PM2.5 exposure. Betulinic acid derivative inhibited PM2.5-induced acute inflammatory exudate, alveolar septae damage, and inflammatory cell infiltration in lungs of mice.ConclusionsIn the mouse model of PM2.5-induced COPD, betulinic acid derivative reduced the degree of lung inflammation and downregulated inflammatory mediators. These findings support the need for further in vivo studies and clinical evaluation of betulinic acid derivative in patients with COPD.

Exercise and/or Genistein Treatment Impact Gut Microbiota and Inflammation after 12 Weeks on a High-Fat, High-Sugar Diet in C57BL/6 Mice.

Genistein (Gen) and exercise (Exe) have been postulated as potential strategies to ameliorate obesity, inflammation, and gut microbiota (GM) with promising results. However, the impact of the combination of both Exe and Gen is yet to be investigated. We aimed to analyze the impacts of Exe, Gen, and their combined effects on GM and inflammation in mice after a 12-week high-fat, high-sugar diet (HFD). Eighty-three C57BL/6 mice were randomized to control, HFD, HFD + Exe, HFD + Gen, or HFD + Exe + Gen. The V4 region of the 16S rRNA gene was analyzed with Illumina MiSeq. Serum samples were used to analyze interleukin (Il)-6 and Tumor Necrosis Factor alpha (TNF-alpha). The HFD + Exe and HFD + Exe + Gen treatments resulted in significantly greater microbial richness compared to HFD. All the treatments had a significantly different impact on the GM community structure. Ruminococcus was significantly more abundant after the HFD + Exe + Gen treatment when compared to all the other HFD groups. Exe + Gen resulted in serum Il-6 concentrations similar to that of controls. TNF-alpha concentrations did not differ by treatment. Overall, Exe had a positive impact on microbial richness, and Ruminococcus might be the driving bacteria for the GM structure differences. Exe + Gen may be an effective treatment for preventing HFD-induced inflammation.

Human defensins and Th-1 cytokines in hepatitis C viral infection.

Introductionactive or chronic exacerbated forms of hepatitis C virus (HCV) infection subsequently progress to liver disease and human defensins has been determined to have some level of anti-viral properties invitro whilst the expression of T helper-1 cytokines is known to promote complete recovery from acute HCV infection. The study sought to determine relationship between these immune responses.Methodsa cross sectional descriptive study design was employed. Hundred and thirty-two individuals were assessed were assessed for to anti-HCV, HCV RNA, serum levels of human alpha defensins 1 (HAD-1) and human beta defensins 1 (HBD-1). T helper 1 cytokines (IL-2, IFN gamma, TNF alpha) secreted in serum were also analyzed using commercial ELISA assay. The study was conducted in Kumasi, Obuasi and Daboya in Ghana.Resultsthe serum mean concentrations of HAD-1, HBD-1, IL-2, IFN gamma and TNF alpha showed no significant difference in concentrations among participants with chronic, spontaneously recovered or negative to HCV infection (p>0.05). Persons with hepatitis B co-infection were more likely to develop chronic HCV infection (p=0.039). HAD-1 and HBD-1 showed significant positive association with IL-2 (p=0.000) whilst only HAD-1 positively correlated with IL-2 (p<0.000).Conclusionthe immunological markers determined had no association with the status of HCV infection. HAD-1 increased with increasing levels of IL-2. These findings suggest that during HCV infection, inflammatory response through the production of cytokines by IL-2 cells may affect the release of HAD-1 and HBD-1.

Serum biomarkers differentiating Kawasaki disease from febrile infections: A pilot case-control study.

Although some serum biomarkers are elevated in both Kawasaki disease (KD) and infections, these conditions have not been compared by individual or combined biomarkers. The aim of this study, undertaken between January 2016 and May 2018 in a large teaching hospital, was to compare the serum concentration of cytokines, metalloproteinases (MMP) and heat shock protein (HSP) between cases defined as children with Kawasaki disease (KD) and those with febrile infections (controls). Serum concentrations of tumour necrosis factor-alpha (TNF-alpha), interleukins (IL 1beta, 6, and 8), heat shock proteins (HSP 60 and 70) and matrix metalloproteinase (MMP 9) were measured on admission in 17 children under six years of age with a temperature >38.5 °C for ≥five days, and compared between the two groups. The median age was 25 months and the median duration of fever eight days. Seven children were diagnosed with KD and ten had a febrile infection. Only the serum concentrations of IL-6 and TNF-alpha were significantly higher in the former than in the latter group (P = 0.01 and 0.04 respectively). To differentiate between the two groups with the best sensitivity and specificity, the optimal cut-off value for IL-6 was 12.6 pg/mL, and for TNF-alpha 47.9 pg/mL. Their combined increase, however, outperformed their individual concentrations. The characteristic diagnostic “signature” of the combined elevation of IL-6 and TNF-alpha serum levels has the potential, in febrile children, to differentiate early KD from febrile infections, allowing the institution of appropriate therapy.

HMGB1 and inflammatory cytokines in experimental acute lung injury induced in rabbits

ABSTRACT The aim of this work was to measure HMGB1, TNF-alpha, and IL-8 in bronchoalveolar lavage (BAL), serum and TLR2 and TLR4mRNA expression in lung tissue of rabbits with two grades of acute lung injury (ALI). The animals were randomly assigned to groups with severe (S) and mild/moderate (MM) ALI, induced with warm saline, and a control group. HMGB1, TNF-alpha, IL-8, TLR2mRNA and TLR4mRNA were measured after ALI induction. The results showed increased levels of IL-8, TNF-alpha, HMGB1 and TLR4mRNA in the ALI groups. HMGB1, IL-8 and TNF-alpha concentrations in BAL were higher in S compared MM. Increased TLR4mRNA was observed in S and MM versus control. The results suggest an early participation of HMGB1 in ALI together with IL-8 and TNF-alpha and association with severity. TLR4 has early expression and role in ALI pathophysiology but is not associated with severity.

Commentary: Obesity: The “Achilles heel” for COVID-19?

Commentary: Obesity: The “Achilles heel” for COVID-19?

Are adipokines associated with atrial fibrillation in type 2 diabetes?

The potential effect of adipokines on the development of AF is yet to be established. The aim of this study was to investigate the association of baseline serum adipokines with 1) the presence of AF at baseline and 2) future risk of AF development. The current study is a sub-analysis of the prospective, randomised AVOCADO (Aspirin Vs./Or Clopidogrel in Aspirin-resistant Diabetics inflammation Outcomes) trial. The AVOCADO study included patients with type 2 DM burdened with at least two additional cardiovascular risk factors and receiving acetylsalicylic acid. In patients included in the current analysis adipokines and inflammatory biomarker levels were measured. Information on the subsequent AF diagnosis was collected after a median of 5.4 years of follow-up. A total of 273 patients with type 2 DM (median age 68 years; 52% male) were included in the initial analysis comparing patients with and without AF at baseline. Patients with diagnosed AF (12%) had higher levels of serum resistin [8.5 (5.8-10.5) vs. 6.9 (5.6-8.7) ng/mL; p = 0.034], adiponectin [6.9 (5.6-8.7) vs. 2.7 (1.8-4.2) ng/mL; p = 0.032], and N-terminal pro-B-type natriuretic peptide [336 (148-473) vs. 108 [45-217]; p < 0.001) than non-AF patients. There were no significant differences in serum leptin, IL-6, and TNF-alpha concentrations between the two groups. From subjects without known AF at study entry, 19% developed AF at follow-up. In logistic regression analysis, baseline adipokine levels did not predict AF development. In type 2 DM, patients with AF have higher resistin and adiponectin concentrations than patients with no AF. None of the studied adipokines proved a predictor of future AF development.

MicroRNAs in bovine milk exosomes are bioavailable in humans but do not elicit a robust pro-inflammatory cytokine response

BackgroundBovine milk exosomes are studied for their roles as bioactive food compounds and as vehicles for drug delivery. Both lines of investigation converge on immune function, e.g., immune regulation by absorption of microRNAs encapsulated in milk exosomes across species boundaries, and the possibility of exosomes and their cargos triggering an immune response if used in drug delivery. This study assessed the bioavailability of immune-related microRNAs from bovine milk and changes in plasma cytokine concentrations after milk consumption in humans, and the secretion of cytokines by human peripheral blood mononuclear cells (PBMCs) cultured with milk exosomes transfected with immune-relevant microRNAs.ResultsHuman plasma samples were collected before and at timed intervals after a milk meal and analyzed for concentrations of six immune-relevant microRNAs and nine cytokines. The peak plasma concentrations of miR-15b-5p, miR-21-5p, miR-106b-5p, and miR-223-3p were 60 ± 9.80% to 162 ± 31.80% higher after milk consumption (Ct values 23 ± 1.2 to 26 ± 1.1 cycles) compared to baseline values (P < 0.05). Plasma concentrations of TNF-alpha were not significantly different before versus after milk consumption; eight other cytokines were below detection limit. PBMCs were collected before and six hours after milk consumption and cultured with or without concanavalin A (ConA). TNF-alpha, IL-1β, IL-6 and IL-10 were detectable in culture media, but concentrations did not depend on milk consumption prior to PBMC isolation (P > 0.05). When PBMC cultures from fasted subjects were supplemented with milk exosomes that had been transfected with immune-relevant microRNAs, the concentrations of IL-1β, IL-6, IL-10 and TNF-alpha were 29 ± 12% to 220 ± 33% higher than controls cultured with non-transfected exosomes (P < 0.05), but cytokine concentrations were not different compared with control exosomes transfected with scrambled microRNA (P > 0.05).ConclusionsMicroRNAs in bovine milk exosomes are bioavailable. Milk exosomes do not elicit an increase of plasma cytokines following oral administration.Trial registrationISRCTN registry ID: 16329971. Retrospectively registered on February 7th, 2019.

Protective effect of alpha-lipoic acid and omega-3 fatty acids against cyclophosphamide-induced ovarian toxicity in rats.

Background and Aim:Cyclophosphamide therapy is known to be associated with the risk of female infertility as a result of ovarian toxicity. Alpha-lipoic acid (LA) and omega-3 fatty acids are known for their antioxidant and anti-inflammatory activities. The present study investigated the potential protective effect of alpha-LA, omega-3 fatty acids, and its combination against cyclophosphamide-induced ovarian toxicity in rats.Materials and Methods:Thirty rats were equally divided into Groups I, II, III, IV, and V. Group I was normal control, wherein the rats were fed with normal feed and water ad libitum. Group II served as cyclophosphamide-induced group, wherein the rats were injected with cyclophosphamide at 75 mg/kg through intraperitoneal route once a week to induce ovarian toxicity. Groups III and IV were treated with alpha-LA at the rate of 25 mg/kg and omega-3 fatty acids at the rate of 400 mg/kg, respectively, in parallel to cyclophosphamide induction as in Group II. Group V animals were coadministered with alpha-LA (25 mg/kg) and omega-3 fatty acids (400 mg/kg) along with cyclophosphamide induction as in Group II. The respective treatments were administered daily through oral route for a period of 30 days. Regularity of estrous cycle was evaluated by vaginal cytology. Post-treatment period, the animals were humanely sacrificed, and the blood samples were subjected to the estimation of follicle-stimulating hormone (FSH) and estrogen. The ovarian tissue was weighed and subjected to histopathology, transmission electron microscopy, estimation of decreased glutathione (GSH), and tumor necrosis factor (TNF)-alpha.Results:Rats treated with cyclophosphamide alone manifested irregularity in estrous cycle, increased FSH, and reduced estrogen levels. The ovaries showed decreased GSH and increased TNF-alpha concentrations. Histopathological and transmission electron microscopic analysis of the ovarian follicles revealed degenerative changes. Administration of alpha-LA and omega-3 fatty acids as well as the combination of both the treatments demonstrated significant normalization of the estrous cycle and antioxidant defense mechanism as well as ameliorated the hormonal profile and histological architecture of the ovarian follicles. However, appreciable synergistic efficacy of the combination therapy (alpha-LA+omega-3 fatty acids) with respect to the monotherapies was not observed in the present study.Conclusion:The efficacy of alpha-LA and omega-3 fatty acids against cyclophosphamide-induced ovarian toxicity could be attributed to its antioxidant and anti-inflammatory activities that prevented the oxidative damage to the ovaries caused by cyclophosphamide. Hence, our findings suggest that dietary supplementation of alpha-LA and omega-3 fatty acids in women receiving cyclophosphamide therapy could carry potential benefits in preventing cyclophosphamide-induced infertility in childbearing women.

High serum concentration of dipeptidyl peptidase 4 at early stage of obesity – preliminary study

Background: Adipose tissue has been recognized as an endocrine organ of considerable complexity, able to secrete adipose-derived factors named adipokines. The secretion of adipokines depends greatly on the volume of body fat, which in turn significantly changes their activity towards a diabetogenic, proinflammatory, and atherogenic pattern. One of the discovered adipokines is dipeptidyl peptidase 4 (DPP4).&lt;br&gt;Objectives: The aim of this preliminary study was to establish an association between serum concentration of DPP4 and obesity at early stage.&lt;br&gt;Material and methods: A total of 32 obese adult volunteers and 40 lean controls were studied. Total cholesterol, triglycerides, HDL (high-density lipoprotein), LDL (low-density lipoprotein) and glucose concentrations were assessed in serum/plasma samples by using commercial tests. Body mass index (BMI) and waist-hip ratio (WHR) were determined. Serum concentrations of DPP4, leptin, visfatin, CRP (C-reactive protein), and TNF-alpha (tumor necrosis factor alpha) were measured using commercial ELISA immunoassay tests.&lt;br&gt;Results: Serum concentrations of DPP4, leptin and visfatin were significantly higher in obese than in lean subjects. The concentration of DPP4 positively correlated with BMI and body mass. Serum CRP and TNF-alpha were increased in obese compared to non-obese, and had a positive correlation with BMI, WHR and body mass.&lt;br&gt;Conclusions: We showed that there is an association between the DPP4, leptin and visfatin concentration in serum and elevated body weight and BMI even at early stage of obesity (I stage of obesity). It suggest the importance of adipose tissue reduction to prevent rise of adipokines levels and further negative metabolic and inflammatory changes.

Circulating Wnt1-inducible signaling pathway protein-1 (WISP-1/CCN4) is a novel biomarker of adiposity in subjects with type 2 diabetes.

Wnt1-inducible signaling pathway protein 1, or cellular communication network factor 4 (CCN4), a member of CCN family of secreted, extracellular matrix associated signaling proteins, recently was validated as a novel adipose tissue derived cytokine. To assess the relationships between circulating CCN4, adipose tissue distribution and function, and chronic low-grade inflammation in subjects with type 2 diabetes. We observed 156 patients with type 2 diabetes and 24 healthy controls. Serum levels of CCN4, hsCRP and alpha1-acid glycoprotein (alpha1-AGP) were measured by ELISA. Serum concentrations of leptin, resistin, visfatin, adipsin, adiponectin, IL-6, IL-8, IL-18 and TNF-alpha were determined by multiplex analysis. Fat mass and distribution was assessed by DEXA. Mean diameter of adipocytes was estimated in samples of subcutaneous adipose tissue. Patients with diabetes had higher levels of circulating CCN4, leptin, resistin, adipsin, visfatin, hsCRP, alpha1-AGP, and IL-6 (all p<0.02). The CCN4 concentration correlated positively with percentage of fat mass in central abdominal area, as well as with leptin, resistin and visfatin levels; negative correlation was found between CCN4 and mean adipocyte diameter. In multiple regression analysis fat mass in central abdominal area was independent predictor for CCN4 concentration. In subjects with type 2 diabetes serum levels of CCN4 are associated with central abdominal fat mass and adipose tissue dysfunction.

Functional ability of neutrophils in patients with ischemic heart disease

Purpose. The study of protein-producing and lipid-releasing ability of neutrophils and its clinical significance in patients with IHD.Materials and methods. 25 patients with arterial hypertension without clinical and ultrasound manifestations of atherosclerosis were examined; 47 patients with coronary artery disease with stable angina, as well as 19 patients with unstable primary angina. The comparison group — 33 healthy persons. In the serum, enzyme-linked immunosorbent assay was used to evaluate the concentration of C-reactive protein (CRP), lipoprotein a (LPa), VII coagulation factor VII (VIIf), defensins-alpha (1–3), interleukin-6 (IL-6), interleukin-8 (IL-8), tumor necrosis factor alpha (TNF-alpha). Leukocytes, predominantly represented by neutrophils, which were further cultivated, were excreted from peripheral blood in the presence of dextrans; the levels of CRP, LPa, defensins-alpha (1–3), VIIf, and lipid-releasing ability of leukocytes (LRAL) were assessed according to the method of A. V. Tuev and V. Yu. Mishlanov. An angiographic study, duplex scanning arteries, and echocardiography were performed.Results. It has been established that in patients at the stage of preclinical atherosclerotic lesion, the ability of leukocytes to form protein-lipid complexes (PLC), marked by LRAL, practically does not differ from healthy individuals, but already at this stage of atherogenesis an increase in their ability to produce antimicrobial peptides — defensins-alpha (1–3). In patients with coronary artery disease with stable angina pectoris, an increase in leukocyte production of a wide range of proteins — defensins-alpha (1–3), VIIf, LPA and CRP in combination with signs of a systemic inflammatory reaction, marked by high concentrations of CRP, interleukin-6 and tumor necrosis factor alpha in serum, was noticed. At this stage of atherogenesis, the value of LRAL is significantly different from the values of healthy individuals. It is shown that the higher the LRAL value, the higher the functional class of chronic heart failure in these patients (R = 0.4; p = 0.02). In patients with coronary artery disease with primary unstable angina, the maximum LRAL value was detected, and in patients with ultrasound signs of plaque instability, high concentrations of CRP in neutrophilic supernatants were found. Correlation analysis revealed that, in patients with coronary artery disease, the average percentage of coronary artery stenosis, according to angiography, is directly related to the LRAL value (R = 0.7; p = 0.03) and the serum concentration of TNF-alpha (R = 0.5; p = 0.03), and the number of clinically significant stenoses — with the magnitude of LRAL (R = 0.4; p = 0.04) and the concentration in supernatants VIIf (R = 0.5; p = 0.04).Conclusion. The LRAL value, CRP and defensin-alpha content in leukocyte supernatants in IHD patients are interconnected with the severity and activity of atherosclerotic lesion of arteries, which is confirmed by the results of examination of patients at the stage of preclinical, clinically manifest stable and unstable atherosclerotic lesion.

Circulating Leptin, Adiponectin, and Tumor Necrosis Factor-Alpha in Patients Undergoing Surgery Due to Colorectal Cancer

Background: Adipocytokines have been proposed as factors mediating associations between obesity and inflammation in patients with colorectal cancer (CRC). Thus, the aim of this study was to determine the clinical relationships between blood concentrations of leptin (LEP), adiponectin (ADP), and tumor necrosis factor alpha (TNF-alpha) and the outcomes measured in patients with CRC undergoing surgery. Patients and Methods: History, body composition, and blood concentrations of LEP, ADP, and TNF-alpha were determined in 107 patients undergoing surgery due to CRC. The patients were followed up for 619.72 ± 371.65 days. Results: Compared to patients with stage II CRC, individuals with clinical stage I CRC had significantly lower ADP and higher TNF-alpha blood concentrations. We found significant correlations between the clinical stage of CRC (early vs. localized vs. metastatic) and the following: crude blood ADP concentration (R = 0.25; p = 0.015), ADP-to-TNF-alpha ratio (R = 0.31; p = 0.002), and ADP when indexed to body surface area (R = 0.25; p = 0.008) and to fat mass (R = 0.25; p = 0.016). The risk of death during the long-term follow-up period was independently related to the clinical stage of CRC, impairment of the patient’s functional status, and higher blood carcinoembryonic antigen concentration. In Kaplan-Meier survival analysis, patients with blood LEP concentrations adjusted to a visceral adipose tissue score of ≥0.47 had a significantly better likelihood of surviving than their counterparts. Conclusions: In patients with CRC undergoing surgery, blood ADP and TNF-alpha concentrations were associated with the clinical stage of the cancer, likelihood of radical tumor excision, occurrence of nonsurgical postoperative complications, and long-term survival, which suggests the role of dysregulation in the endocrine function of adipose tissue in response to the neoplasmatic process.

Abstract #4294 Plasma concentrations of CRP, IL-6, IL-10, and TNF-alpha in patients with depression before and after treatment with electroconvulsive therapy

Abstract #4294 Plasma concentrations of CRP, IL-6, IL-10, and TNF-alpha in patients with depression before and after treatment with electroconvulsive therapy

Pharmacokinetics of Recombinant Human Tumor Necrosis Factor Alpha in the Delivery System

The main problems of using TNF-alpha in antitumor therapy are its rapid degradation in the bloodstream and the limited selectivity of accumulation in the tumor tissue. The SRC VB «Vector» developed a biodegradable molecular construct that provides protection against proteases and ensures targeted delivery of proteins to the tumor tissue. This construct was used to create an antitumor drug containing recombinant human TNF-alpha (rhTNF-alpha).The aim of the study was to analyse rhTNF-alpha pharmacokinetics in the delivery system after a single administration.Materials and methods: the rhTNF-alpha drug carried by the delivery system was intravenously administered to female outbred ICR (СD-1) mice only once at two effective antitumor doses, 2.55 μg and 5.1 μg / 20 g of body weight. The concentration of TNF-alpha in the serum and supernatants of organ homogenates, obtained at different time points after administration, was analysed by immunoenzyme assay.Results: the obtained curves of TNF-alpha concentration in the blood were satisfactorily described by the equation for the twocompartment model without absorption. The rapid phase of elimination from the blood took 0–4 h, the slow one — 4–24 h. The highest specific content of protein was observed in the skin, spleen, and kidneys tissue. The calculation of pharmacokinetic parameters demonstrated that the highest values of tissue availability fT were obtained for the kidneys and skin; the drug was retained for longer periods of time in the kidneys, liver and skin (according to the MRT data). As a rule, complete elimination of the drug was observed by the end of the first day after administration.Conclusions: rhTNF-alpha carried by the delivery system was quickly eliminated from the blood and distributed in the internal organ tissues after a single intravenous administration to mice in the effective doses range. The main organs in which rhTNF-alpha was distributed were skin, kidneys, and spleen. The elimination of the drug from the blood was a two-phase process which was generally over by the end of the first day.

FOXP3+ T-REGULATORY LYMPHOCYTES IN HYPERTENSIVE PATIENTS

Objective: Development of coronary atherosclerosis in patients with arterial hypertension (AH) is associated with the state of chronic low-grade inflammation, but the role and function of T-regulatory (Treg) lymphocytes in this process is not yet fully elucidated. Design and method: In total 24 patients with AH of high and very high cardiovascular risk (8 men and 16 women; 61.0 (55.5; 64.00) y.o.) have been recruited in the study. All the patients underwent coronary angiography to verify the presence of coronary stenosis. Among all the recruited patients 16 individuals were characterized by at least 50% coronary stenosis. Numbers of Treg-lymphocytes were evaluated by flow cytometry according to the expression of CD4, CD25 and FoxP3 molecules. Concentrations of high-sensitive C-reactive protein (hsCRP) were measured by ELISA. Concentrations of inflammatory and anti-inflammatory cytokines were evaluated in serum and in supernatants of intact and lipopolysaccharide (LPS)-stimulated peripheral blood mononuclear cells’ culture by multiplex analysis. Results: Patients with coronary stenosis higher than 50% (Gr. 1) were characterized by elevated numbers of CD4+FoxP3+ Treg lymphocytes compared to the patients with minor coronary changes (Gr. 2) (3.67 (2.61; 5.37) % vs. 2.33 (1.56; 3.68) %; p = 0.047), as well as by elevated inflammatory markers, such as hsCRP (6.24 (1.98; 6.95) mg/l in Gr. 1 vs. 1.79 (1.32; 2.51) mg/l in Gr. 2; p = 0.066) and LPS-stimulated secretion of TNF-alpha (9126 (6908; 14108) pg/ml in Gr. 1 vs. 2517 (2512; 6198) pg/ml in Gr. 2; p = 0,030). Both parameters are typical to the chronic low-grade inflammation. In total group of patients concentration of serum TNF-alpha was directly related to the concentration of fasting and postprandial glucose (Rs = 0,536; p = 0,032 and Rs = 0,726; p = 0,001, respectively) as well as to the levels of HbA1c (Rs = 0,588; p = 0,035). We have revealed inverse relationships between numbers of CD25hiFoxP3+ Treg-lymphocytes and fasting levels of insulin (Rs = -0,565; p = 0,035) and HOMA (Rs = -0,666; p = 0,009). Conclusions: We have revealed the increase of Treg-lymphocytes’ numbers in patients characterized with 50% stenosis of coronary arteries, which may have compensatory role to counterbalance the state of the chronic low-grade inflammation and may be associated with the poor glycemic control in these patients.

Role of adipose tissue inflammation in fat pad loss induced by fasting in lean and mildly obese mice

Inflammation induced by obesity contributes to insulin resistance and atherosclerosis. Indeed, high levels of proinflammatory cytokines trigger chronic low-grade inflammation and promote detrimental metabolic effects in the adipose tissue. On the other hand, inflammation seems to control fat pad expansion and to have important functions on lipolysis and glucose metabolism. Thus, it is possible that inflammation may also drive fat pad loss, as seen during long-fast periods. Herein, we have used fasting as a strategy to induce weight loss and evaluate the possible role of inflammation on adipose tissue remodeling. Male BALB-c mice were fed with chow diet (lean mice) or with high-carbohydrate refined diet (mildly obese mice) for 8 weeks. After that, animals were subjected to 24 h of fasting. There was a 63% reduction of adiposity in lean mice following fasting. Furthermore, the adipose tissue was enriched of immune cells and had a higher content of IL-6, TNF-alpha, IL-10, TGF-β and CXCL-1. Interestingly, mildly obese mice, subjected to the same 24-h fasting period, lost only 33% of their adiposity. Following fasting, these mice did not show any increment in leukocyte recruitment and cytokine levels, as did lean mice. Our findings indicate that inflammation participates in fat mass loss induced by fasting. Although the chronic low-grade inflammation seen in obesity is associated with metabolic diseases, a lower inflammatory response triggered by fasting in mildly obese mice impairs fat pad mobilization.

A preliminary study assessing cytokine production following ex vivo stimulation of whole blood with diet in dogs with chronic enteropathy

BackgroundEx vivo whole blood stimulation assays (WBSA) have been used to characterize the cytokine response to diet in cats. The present study aimed to use this assay to determine the cytokine response to diets being fed at the time of diagnosis to dogs with chronic enteropathy (CE) and to compare this to a control group of dogs presented for non-gastrointestinal (GI) causes.ResultsDogs with chronic GI signs and dogs presented for non-GI causes were prospectively recruited. For each case, residual blood following diagnostic sampling was placed into heparin. WBSAs were performed using crude extracts of the diet currently being fed and provided by the owner. Supernatants were collected and analyzed for tumor necrosis factor (TNF)-alpha, interleukin (IL)-10 and IL-4 using an enzyme-linked immunosorbent assay. The case group consisted of 22 dogs with CE diagnosed on histopathology of GI biopsy and 9 with suspected CE. The non-GI group consisted of 18 dogs. Of the diets being fed at or prior to diagnosis, hydrolyzed protein diets elicited significantly lower IL-10 and TNF-alpha concentrations compared to commercial intact protein diets in dogs with confirmed or suspected CE (P-value 0.004 and < 0.001, respectively). Six out of 15 dogs with detectable IL-4 concentrations in the confirmed CE group had IL-4 to IL-10 ratios that exceeded the 95% confidence interval (CI) of the mean of the non-GI group (non-GI: 95% CI of IL-4:IL-10 = 0.64–2.71; confirmed CE: IL-4:IL-10 in 6 dogs = mean 22.40, range 2.77–89.11).ConclusionsHydrolyzed protein diets elicited a significantly reduced cytokine response when incubated with patient whole blood ex vivo compared to commercial intact protein diets in dogs with CE. The IL-4 to IL-10 ratio as a marker of dietary responsiveness warrants further investigation, together with assessment of the cytokine response to diet at the intestinal mucosal surface.

High Serum Uric Acid Is Associated with Tubular Damage and Kidney Inflammation in Patients with Type 2 Diabetes.

Background Uric acid presents different roles in an organism. High serum uric acid concentrations may induce inflammatory pathways and promote kidney damage through different mechanisms. Therefore, this study investigated the association among high serum uric acid concentrations, renal tubular damage, and renal inflammation assessed via estimation of urinary kidney injury molecule-1 (KIM-1) and inflammatory cytokines in patients with type 2 diabetes (T2D). Methods Urinary concentrations of KIM-1, IL-1, IL-6, IL-10, and TNF-alpha, as well as other biochemical parameters, were assessed in 125 patients with T2D who were grouped into two groups based on the serum uric acid levels (<6.0 mg/dL and ≥6.0 mg/dL). Patients were also stratified according to the tertiles of serum uric acid concentrations. Results Urinary KIM-1, IL-1, IL-6, and TNF-alpha were higher in patients with serum uric acid concentrations ≥ 6.0 mg/dL. However, the differences between the groups were not statistically significant when the urinary values of KIM-1 and cytokines were normalized by the urinary creatinine concentration. Serum uric acid concentrations were significantly associated with urinary KIM-1 (values normalized by urinary creatinine concentration) and urinary TNF-alpha (absolute values and values normalized by urinary creatinine concentration), independent of the body mass index (BMI) and estimated glomerular filtration rate (eGFR). Conclusions High serum uric acid concentrations were associated with high urinary KIM-1 levels accompanied by the increase of urinary proinflammatory cytokines in patients with T2D. However, normalization of urinary markers by urine creatinine concentration seems to influence the profile of the results.