Concentrations Of Tumor Necrosis Factor Research Articles

Tumor necrosis factor-α, TNF receptor, and soluble TNF receptor responses to aerobic exercise in the heat

The purpose of this study was to evaluate the effects of aerobic exercise in the heat on circulating concentrations of tumor necrosis factor (TNF)-α, soluble TNF receptors (STNFR1&2), and surface expression of TNFR1&2 on monocyte subpopulations. Twelve recreationally active Caucasian men (24.4 ± 3.4 yrs.; 180.0 ± 6.8 cm; 81.5 ± 8.0 kg; 47.2 ± 4.8 mL·kg−1·min−1) completed an exercise protocol in three environmental conditions: high temperature/low humidity [HTLH; 35 °C, 20% relative humidity (RH)]; high temperature/moderate humidity (HTMH; 35 °C, 45%RH); and moderate temperature/moderate humidity (MTMH; 22 °C, 45%RH). Each protocol consisted of a 60-minute cycling trial at 60% VO2max, a 15-minute rest, and a time-to-exhaustion trial at 90% VO2max (TTE). Blood was sampled before (PRE), immediately after (POST) the 60-minute trial, immediately post-TTE (PTTE), and one-hour post-TTE (REC). Circulating TNF-α and STNFR1&2 were assayed. TNFR1&2 expression on monocyte subsets was measured by flow cytometry on a subset of participants (n = 8). TNF-α area under the curve with respect to increase (AUCi) was greater during HTMH compared to MTMH and HTLH. STNFR1 concentration was greater during HTMH compared to MTMH. With all trials combined, STNFR1 concentration increased from PRE to POST, PTTE, and REC. TNFR1 expression on non-classical monocytes was greater during HTMH compared to HTLH while TNFR2 expression was lower during HTLH compared to both MTMH and HTMH. Data suggest that exercise in the heat increases circulating TNF-α and STNFR1 concentration concomitantly. Furthermore, non-classical monocyte expression of TNFRs are impacted by temperature and humidity during exercise.

Subthalamic Deep Brain Stimulation Affects Plasma Corticosterone Concentration and Peripheral Immunity Changes in Rat Model of Parkinson\u2019s Disease

Deep brain stimulation of the subthalamic nucleus (DBS-STN) is an effective treatment for advanced motor symptoms of Parkinson’s disease (PD). Recently, a connection between the limbic part of the STN and side effects of DBS-STN has been increasingly recognized. Animal studies have shown that DBS-STN influences behavior and provokes neurochemical changes in regions of the limbic system. Some of these regions, which are activated during DBS-STN, are involved in neuroimmunomodulation. The therapeutic effects of DBS-STN in PD treatment are clear, but the influence of DBS-STN on peripheral immunity has not been reported so far. In this study, we examined the effects of unilateral DBS-STN applied in male Wistar rats with 6-hydroxydopamine PD model (DBS-6OHDA) and rats without nigral dopamine depletion (DBS) on corticosterone (CORT) plasma concentration, blood natural killer cell cytotoxicity (NKCC), leukocyte numbers, lymphocyte population and apoptosis numbers, plasma interferon gamma (IFN-γ), interleukin 6 (IL-6), and tumor necrosis factor (TNF-α) concentration. The same peripheral immune parameters we measured also in non-stimulated rats with PD model (6OHDA). We observed peripheral immunity changes related to PD model. The NKCC and percentage of T cytotoxic lymphocytes were enhanced, while the level of lymphocyte apoptosis was down regulated in 6OHDA and DBS-6OHDA groups. After DBS-STN (DBS-6OHDA and DBS groups), the plasma CORT and TNF-α were elevated, the number of NK cells and percentage of apoptosis were increased, while the number of B lymphocytes was decreased. We also found, changes in plasma IFN-γ and IL-6 levels in all the groups. These results suggest potential peripheral immunomodulative effects of DBS-STN in the rat model of PD. However, further studies are necessary to explain these findings and their clinical implication.Graphical Influence of deep brain stimulation of the subthalamic nucleus on peripheral immunity in rat model of Parkinson’s disease.

Characteristics of autologous protein solution and leucocyte-poor platelet-rich plasma for the treatment of osteoarthritis of the knee

Recently, platelet-rich plasma (PRP) has received attention as a treatment for patients with osteoarthritis of the knee (OAK), a chronic degenerative disease, to bridge the gap between conservative and surgical treatments. Here, we investigated the differences in the humoral factors present in two types of PRP purified using the Autologous Protein Solution (APS) kit (group Z; leucocyte-rich PRP) or the Cellaid Serum Collection Set P type (group J; leucocyte-poor [LP]-PRP). Differences in humoral factors between healthy subjects (n = 10) and OAK patients (n = 12; group Z = 6, group J = 6), and the relationship between humoral factors and clinical outcome scores were investigated. Both anti-inflammatory and inflammatory cytokines were highly enriched in APS. The concentrations of tumour necrosis factor (TNF)-α, platelet-derived growth factor, fibroblast growth factor, soluble TNF-receptor 2, soluble Fas and transforming growth factor-β1 were higher in group Z, while the total amounts were higher in group J. The concentration of interleukin-1 receptor antagonist was positively correlated with the magnitude of change in the clinical outcome score and may contribute to improving knee-joint function. This is the first description of the humoral factors in APS and LP-PRP prepared from healthy subjects or OAK patients of Asian descent.

Bu-Shen-Fang-Chuan formula attenuates cigarette smoke-induced inflammation by modulating the PI3K/Akt-Nrf2 and NF-κB signalling pathways

Ethnopharmacological relevanceChronic obstructive pulmonary disease (COPD) is a respiratory inflammatory disease. Unlike asthma, COPD is insensitive to glucocorticoid treatment; thus, it is of great importance to find alternative medications, including Chinese medicine, to suppress inflammation. Bu-Shen-Fang-Chuan formula (BSFCF) is commonly used for the treatment of COPD in China. However, the mechanisms of BSFCF in COPD treatment are still unclear. Aim of the studyTo verify the anti-inflammatory efficacy of BSFCF in COPD and to explore the possible mechanisms underlying its anti-inflammatory efficacy based on the phosphatidylinositol 3 kinase (PI3K)/protein kinase B (Akt)-Nuclear factor erythroid 2-related factor 2 (Nrf2) and Nuclear factor (NF)-κB signalling pathways. Materials and methodsA rat model of COPD was established by chronic exposure to cigarette smoke (CS) for 6 months. Bronchoalveolar lavage fluid (BALF) and blood were obtained to detect inflammatory cytokines. Lung samples were harvested, and part of each sample was fixed for subsequent H&E staining and immunohistochemical (IHC) analysis. The remaining lung tissues were used for RNA sequencing analysis and western blotting. ResultsBSFCF significantly reduced inflammatory infiltration in the lungs of CS-exposed rats and decreased the concentrations of tumor necrosis factor (TNF)-α and interleukin (IL)-6 in both the BALF and serum. Additionally, BSFCF evidently attenuated NF-κB activation and downregulation of glucocorticoid receptor (GR) caused by CS. Furthermore, BSFCF increased the activation of PI3K/Akt-Nrf2 signalling in response to CS. ConclusionsBSFCF attenuated CS-induced inflammation in COPD, which was partially achieved through the PI3K/Akt-Nrf2 and NF-κB signalling pathways.

4221 Lower Serum TWEAK Concentration is a Biomarker for Mortality in Community Acquired Pneumonia

OBJECTIVES/GOALS: To determine the relationship among serum concentration of tumor necrosis factor (TNF)-like weak inducer of apoptosis (TWEAK) and mortality in community-acquired pneumonia (CAP) patients. METHODS/STUDY POPULATION: This is a multicenter 2-year cohort study in Spain, designed to better understand the role of sTWEAK concentrations in CAP patients. CAP patients were prospectively enrolled in two University hospitals and sTWEAK was measured within the first 24 hours of ICU admission. Samples were collected and stored for laboratory analyses. To detect sTWEAK in human samples, we used a commercially available ELISA kit following manufacturer’s instructions. Demographic patients’ characteristics and ICU mortality were prospectively collected. Descriptive statistics and logistical regressions were used to assess the proposed aims. RESULTS/ANTICIPATED RESULTS: A total of forty-three patients were included in the study (10 healthy users, 10 uninfected controls and 23 CAP patients). In comparison to healthy volunteers, patients admitted to the hospital (both, infected and non-infected) had lower level of sTWEAK. During hospital admission, 7 (17%) patients died. Patients whom died during ICU stay due to CAP, had significantly lower levels of sTWEAK when comparing with patients whom survived (Median [IQR]; 509.35 [357.49, 953.92] Vs 1103.03 [716.93, 1663.16]; p = 0.015). In contrast, patients that developed shock did not have different concentrations of sTWEAK (Median [IQR]; 1008.04 [531.87, 1390.80] Vs 1062.29 [575.24, 1598.83], p = 0.84). DISCUSSION/SIGNIFICANCE OF IMPACT: Community-acquired pneumonia (CAP) is the first cause of death in underdeveloped countries. CAP is a pulmonary infection that creates a proinflammatory environment not just locally but also systemically, secondary to upregulation of molecular cascades with a wide variety of proteins being released perpetuating this inflammation and tissue damage. Several of these molecules have been described and linked to a greater risk of inhospital complications, longer length of hospital stay and mortality. TNF-like weak inducer of apoptosis (TWEAK) is a member of the TNF-alpha superfamily, involved in immune response, cell growth, angiogenesis, NF-kB activation and apoptosis induction in tumor cells. It is known that serum-TWEAK plays a role in inflammatory processes, however, its behavior is unknown in patients with CAP. Therefore, this study aims to identify whether there is a relationship between serum concentration of TWEAK and prognosis in CAP patients. To our knowledge, this is the first study to shown that concentration of sTWEAK within the first 24 hours of ICU admission is lower in patients with CAP. Moreover, patients whom died during ICU admission due to CAP, have lower sTWEAK levels. This biomarker may identify patients at higher risk of dying due to CAP and may represent severe CAP. However, further studies are needed to confirm these findings.

О влиянии нового производного пиримидина PIR-4 на концентрацию маркеров внешнего и внутреннего путей апоптоза в условиях экспериментальной церебральной ишемии

Исследовано антиапоптотическое действие нового производного пиримидина под шифром PIR-4 в условиях экспериментальной фокальной церебральной ишемии у крыс линии Вистар, разделенных на 5 групп (n = 10). В первую группу включены ложнооперированные особи, вторая группа — крысы негативного контроля, получавшие взвесь очищенной воды и твина-80. Третья и четвертая группы представлены животными, получавшими препараты сравнения кавинтон (3,2 мг/кг) и глиатилин (60 мг/кг), соответственно. Пятой группе крыс вводили соединение PIR-4. Все соединения вводили сразу после моделирования ишемии и в течение 3 сут ежедневно. Модель фокальной ишемии мозга — окклюзия левой средней мозговой артерии (наркоз — хлоралгидрат, 350 мг/кг). В ходе проведенного исследования установлено, что на фоне моделируемой патологии возрастает концентрация фактора некроза опухоли (TNF-α), фактора, индуцирующего апоптоз (AIF), p53-зависимого модулятора апоптоза (PUMA), снижается уровень Jun-N-концевой киназы (JNK). Препараты сравнения кавинтон и глиатилин корректировали возникшие изменения, препятствуя развитию внешнего и внутреннего путей апоптоза. На фоне получения крысами вещества PIR-4 также уменьшал концентрацию маркеров каспаза-зависимого и каспаза-независимого путей апоптоза, при этом по антиапоптотическому эффекту, превосходя кавинтон и не уступая глиатилину.

Maternal Vitamin D Sufficiency Is Associated with Lower Circulating TNF-α in Breastfeeding Infants

Abstract Objectives Exclusively breastfeeding (BrF) infants are at risk for vitD deficiency if they or their mothers are not supplemented. However, it is unknown if maternal or direct infant vitD supplementation could have differential effects on the BrF child. We hypothesized that maternal vitD status may affect concentrations of pro-inflammatory cytokines in infants. Methods Exclusively BrF infants and mothers were enrolled in a clinical study in which infant vitD sufficiency was achieved by either direct supplementation with 400 IU/d vitD3 or mothers were supplemented with 6400 IU/d. Infant and maternal plasma 25-hydroxyvitamin D (25-D) values were determined by radioimmunoassay at enrollment (1-mo infant) and completion (5-mo). Concentrations of 10 pro-inflammatory cytokines in infant plasma were determined by electrochemiluminescence. ANOVA, t-tests and linear correlations were used to calculate relationships between vitD and circulating cytokines in 49 infant-mother pairs at study enrollment and 39 pairs at completion. Results Plasma concentrations of 10 pro-inflammatory cytokines measured were very low (pg/ml range). Neither infant nor maternal vitD status (sufficiency defined as plasma 25-D > 30 ng/ml) were associated with 9 measured cytokines in infant blood (IFN-γ, IL-1b, –2, –4, –6, –8, –10, –12, –13). In contrast, concentrations of tumor necrosis factor (TNF)-α in infant plasma were inversely correlated to maternal vitD status (r = −0.37; P = 0.02 at study completion). At study enrollment, BrF infants of vitD-sufficient mothers had mean 20% lower plasma TNF-α concentration than infants of vitD-insufficient mothers (P = 0.02), regardless of the infant's own vitD status. VitD sufficiency in infants per se was associated with mean 5% lower plasma TNF-α concentration, non-significantly different to that of vitD-deficient infants. Conclusions These results indicate that vitD-mediated changes in breastmilk composition can affect the immune system of BrF infants. Circulating concentrations of the major pro-inflammatory cytokine TNF-α were significantly lower in infants with vitD-sufficient mothers regardless of the infant's own vitD status. Studies are currently investigating vitD-associated effects on responses of BrF infants’ immune cells to antigenic challenge. Funding Sources NIH/NCATS, SC Translational Research Institute, MUSC Pediatrics.

Ulinastatin alleviates mitochondrial damage and cell apoptosis induced by isoflurane in human neuroglioma H4 cells.

Isoflurane has been demonstrated to induce mitochondrial damage and cell apoptosis. The isoflurane-induced inflammation may be an important reason for this phenomenon. Studies have shown that ulinastatin (UTI) has an anti-inflammatory effect. Our aim was to investigate whether UTI could attenuate isoflurane-induced mitochondrial damage and cell apoptosis by inhibiting inflammation. Human neuroglioma H4 cells were exposed to isoflurane with or without UTI. The ratio of cell apoptosis was evaluated by flow cytometry. β-Amyloid (Aβ) peptide and cleaved caspase 3 expression were evaluated by Western blot analysis. The concentrations of tumor necrosis factor α (TNF-α) and interleukin 1β (IL-1β) were detected by sandwich enzyme-linked immunosorbent assays. Mitochondrial structural changes were detected by transmission electron microscopy. Mitochondrial membrane potential (Δψm) was determined by 5,5',6,6'-Tetrachloro-1,1',3,3'-tetraethyl-imidacarbocyanine iodide (JC-1). The activity of the mitochondrial electron transport chain (ETC) complexes I, II, III, and IV was determined by assay kits. UTI attenuated the TNF-α and IL-1β release induced by isoflurane. UTI could also reduce mitochondrial structure damage, mitigate the decrease in Δψm, and improve ETC complexes dysfunction. Furthermore, it decreased cell apoptosis induced by isoflurane in H4 cells. UTI had no effect on isoflurane-induced Aβ expression. UTI may mitigate isoflurane-induced mitochondrial damage and cytotoxicity by inhibiting inflammation.

Protective effects of dietary mannan oligosaccharide on heat stress-induced hepatic damage in broilers.

Heat stress is a major concern in broiler's production, which can damage liver of broilers. This study investigated the protective effects of mannan oligosaccharide (MOS) on heat stress-induced hepatic injury in broilers. A total of 144 day-old male chicks were allocated into three treatment groups. Broilers raised under normal ambient temperature were fed a basal diet (control group), and broilers under heat stress (32-33°C for 8h daily) were given the basal diet supplemented without MOS (heat stress group) or with 1g/kg MOS (MOS group) for 42days. Compared with the control group, heat stress reduced liver weight, whereas increased aspartate aminotransferase (AST) and alanine aminotransferase (ALT) activities in the serum. It also reduced glutathione peroxidase (GSH-Px) activity in the serum and liver, GSH content, and superoxide dismutase (SOD) activity in the liver, but increased malondialdehyde (MDA) concentration in the serum and liver. Dietary MOS decreased serum ALT activity in heat-stressed broilers. MOS inclusion also decreased serum MDA content, but elevated hepatic GSH-Px and SOD activities, with MDA content and GSH-Px activity still being different from the control group, and SOD activity being similar to the control group. Heat stress increased concentrations of tumor necrosis factor α (TNF-α) in the serum and liver, interleukin-1β (IL-1β) in the liver, and mRNA abundances of HSP70, TLR4, MyD88, TNF-α, and IL-1β in the liver of broilers. Serum TNF-α content and mRNA abundances of hepatic TLR4 and TNF-α in MOS group were lower than the heat stress group, whereas these indexes were still higher than the control group. Our results indicated that dietary MOS ameliorated hepatic damage in heat-stressed broilers through alleviation of oxidative stress and inflammation.

Histological, metabolic, and inflammatory changes in the renal tissues of high-fat diet-induced obese rats after vitamin D supplementation

PurposeObesity is a major risk factor for chronic renal fibrosis and kidneys’ structural and inflammatory impairments. This study aims to examine the possible therapeutic effects of vitamin D supplementation against renal inflammatory and kidney’s structural fibrosis and degeneration.Design/methodology/approachForty male Wistar rats were divided into two groups for 16 weeks: normal diet (ND) and high-fat diet (HFD); then, each group was subdivided into two groups including ND, ND + vitamin D and HFD, HFD + vitamin D. Vitamin D supplementation was done for five weeks at 500 IU/kg dosage. Renal tissue concentrations of tumor necrosis factor (TNF)-α, interleukin 6, interleukin 1 beta, monocyte chemoattractant protein (MCP)-1 and transforming growth factor-beta (TGF-β), serum values of lipids, markers of glucose homeostasis and urea, creatinine and uric acid and renal tissue histological and structural changes were determined.FindingsHFD feeding caused remarkable histological and structural changes including higher TNF-α, MCP-1 and TGF-βconcentrations in renal tissues of rats, whereas vitamin D has potent anti-inflammatory effects (P= 0.036, 0.047 and 0.02, respectively). Vitamin D administration also reduced urea and uric acid concentrations (P= 0.023 and 0.049, respectively). Moreover, vitamin D reduced glomerulomegaly, reduced lipid accumulation and limited dilated Bowman’s space in rats and improved glycemic status by increasing insulin (P= 0.04) and reducing insulin resistance (P= 0.006).Research limitations/implicationsThe current study has some limitations. It was better to measure the level of inflammatory cytokines’ expression in the kidney tissues. Additionally, the measurement of baseline values of inflammatory cytokines was not possible because of the possibility of animals’ drop-out.Practical implicationsAccording to the study findings, vitamin D treatment in the current report showed a significant therapeutic role in reducing inflammation, improving glycemic and lipid abnormalities and structural and histological modifications in renal tissues of rats. These findings have a great value because after confirming in a human model, vitamin D can be suggested as a potential therapeutic tool in clinical practice.Social implicationsAfter being confirmed by other animal or human researches, the results of the current work could have great social implications by reducing the prevalence of obesity-related renal complications and highlighting the beneficial roles of vitamin D.Originality/valueTo the best of the authors’ knowledge, this is the first study to investigate the histological and inflammatory changes in the kidneys and metabolic parameters in the HFD induced rats and also clarified the therapeutic roles of vitamin D in ameliorating the inflammatory, histological, metabolic and functional changes in the kidneys of obese rats.

MiR-599 regulates LPS-mediated apoptosis and inflammatory responses through the JAK2/STAT3 signalling pathway via targeting ROCK1 in human umbilical vein endothelial cells.

MicroRNA plays an integral role in the development of atherosclerosis. Our study aimed to investigate the roles of miR-599 in lipopolysaccharide (LPS)-induced endothelial damage in human umbilical vein endothelial cells (HUVECs). HUVECs were transfected with a miR-599 mimic and negative control, and then exposed to LPS. The expression of miR-599 was detected by quantitative real time-polymerase chain reaction (RT-qPCR). Cell viability was analyzed by CCK-8 assay and trypan blue exclusion assay; the formation of DNA fragments was tested by Cell Death Detection ELISA Plus kit; the incidence of apoptosis was detected by flow cytometry; the expression of p53 and cleaved-caspase 3 (c-caspase 3) was evaluated by western blot. Moreover, the mRNA levels and concentrations of tumour necrosis factor (TNF)-α, interleukin (IL)-6, ICAM-1 and VCAM-1 were assayed by RT-qPCR and ELISA. The results showed that overexpression of miR-599 increased cell viability, reduced DNA fragments, the incidence of apoptosis, as well as the protein levels of p53 and c-caspase 3 in the presence of LPS. TNF-α, IL-6, ICAM-1 and VCAM-1 mRNA levels and concentrations were also decreased upon miR-599 upregulation. In addition, the dual luciferase reporter assay demonstrated that ROCK1 is a direct target of miR-599. MiR-599 overexpression inhibited ROCK1 expression. Induced expression of ROCK1 reversed the roles of miR-599 in apoptosis and inflammation. The gain function of miR-599 function inhibited activation of the JAK2/STAT3 signalling pathway, which was abrogated by overexpression of ROCK1. Taken together, our results indicate that miR-599 attenuates LPS-caused cell apoptosis and inflammatory responses through the JAK2/STAT3 signalling pathway via targeting ROCK1.

Effects of delayed intraventricular TLR7 agonist administration on long-term neurological outcome following asphyxia in the preterm fetal sheep

In the preterm brain, accumulating evidence suggests toll-like receptors (TLRs) are key mediators of the downstream inflammatory pathways triggered by hypoxia-ischemia (HI), which have the potential to exacerbate or ameliorate injury. Recently we demonstrated that central acute administration of the TLR7 agonist Gardiquimod (GDQ) confers neuroprotection in the preterm fetal sheep at 3 days post-asphyxial recovery. However, it is unknown whether GDQ can afford long-term protection. To address this, we examined the long-term effects of GDQ. Briefly, fetal sheep (0.7 gestation) received sham asphyxia or asphyxia induced by umbilical cord occlusion, and were studied for 7 days recovery. Intracerebroventricular (ICV) infusion of GDQ (total dose 3.34 mg) or vehicle was performed from 1–4 hours after asphyxia. GDQ was associated with a robust increase in concentration of tumor necrosis factor-(TNF)-α in the fetal plasma, and interleukin-(IL)-10 in both the fetal plasma and cerebrospinal fluid. GDQ did not significantly change the number of total and immature/mature oligodendrocytes within the periventricular and intragyral white matter. No changes were observed in astroglial and microglial numbers and proliferating cells in both white matter regions. GDQ increased neuronal survival in the CA4 region of the hippocampus, but was associated with exacerbated neuronal injury within the caudate nucleus. In conclusion, our data suggest delayed acute ICV administration of GDQ after severe HI in the developing brain may not support long-term neuroprotection.

Evaluation of Tumor Necrosis Factor Alpha (TNF‐α) in Sleep‐Deprived Menopausal‐Induced Rats and the Impact on Bone Health

Post‐menopausal osteoporosis as a consequence of estrogen depletion is a growing concern for women in the United States. As more women take on executive positions and experience sleep deprivation, there is the potential for up regulation of pro‐inflammatory cytokines, such as tumor necrosis factor alpha. It follows that the homeostatic imbalance of osteoclastic and osteoblastic activity leads to a greater risk of disease. Bisphosphonates generally, and Zoledronate specifically works by decreasing the number of osteoclasts. This current study investigated the impact of Zoledronate on the concentrations of tumor necrosis factor alpha‐type (TNF‐α) in 32 ovariectomized Wistar rats that were ovariectomized to simulate post‐menopausal estrogen‐deficiency. After one week of surgical recovery, rats were separated into 4 groups. The control group (C, n=4) was given an intraperitoneal (IP) injection of 0.45 mL of 0.9% saline, were housed in standard conditions permitting 12 hours light and 12 hours dark. The Zoledronate group (Z, n=8) were given an IP injection of 50 ug/kg of 10% Zoledronate and housed in standard conditions with a 12 hour light/dark cycle. The sleep‐deprived group (SD, n=8) were given the same IP injection as the rats in C, but were housed in chambers that did not permit sleep for 18 hours, then moved to sleep chambers for 6 hours. The sleep‐deprived Zoledronate group experienced the same sleep deprivation as the SD group, but received an IP injection of 50ug/kg body weight of 10% Zoledronate. Throughout the five‐week period of sleep deprivation cycles, the concentrations of TNF‐α were collected and examined. It was originally hypothesized that the sleep‐deprived group of rats would have the highest concentration of TNF‐α due to the biological stress associated with insomnia. However, TNF‐α levels were significantly higher in the Zoledronate group than both the control and sleep‐deprived groups, as well as the sleep‐deprived with Zoledronate groups (p<0.01). We ascribe this to bisphosphonate induced transient fever seen in Zoledronate usage in previous studies (Dicuonzo, 2003). It is also suspected that the low concentrations of TNF‐α in the sleep‐deprived groups are seen due to the short time frame of this experiment along with a challenged immune system in the animals. With a longer period of sleep deprivation, it is possible that the hypothesized cytokine levels would be reached.Support or Funding InformationThis study was supported in part by Novartis, Basel, Switzerland

Histochemical Effects of Aloe Vera Gel (Aloe Barbadensis Miller) on Puncture-Induced Intervertebral Disc Degeneration in Rabbits

Aim: This study is aimed at investigating the effects of Aloe Vera Gel (AVG) on the histochemical parameters of an annular-punctured rabbit. Methods: Twenty-five rabbits weighing 1.0-3.5 kg were subjected to annular puncture of the L3/L4, L4/L5 and L5/L6 Intervertebral Discs (IVD) using an 18G needle. Rabbits were assigned into five groups (A, B, C, D, and E) with five animals per group. Group A was not punctured. Group B was punctured. Groups C, D, and E were punctured and received AVG. The administration was via oral cannula for 6 weeks. At the end of the experiment, samples were obtained for biochemical and immunochemical analysis with tissue samples excised for histological procedures Results: The results showed a significant increase (F=7.08; P=0.012) in the serum level of Superoxide Dismutase [SOD] and decrease (F=10.20; P=0.0001) in the concentration of Malondialdehyde [MDA]. We found a significant increase in concentrations of both interleukin 1β (F=7.36; P=0.0008) and Tumor Necrosis Factor (TNF) α (F=37.46; P=0.0001). AVG was also able to repair damaged IVD by restoring the histoarchitecture of the nucleus pulposus and annulus fibrosus. Conclusion: We conclude that AVG can repair damaged intervertebral disc and has a strong anti-oxidative and immune modulatory properties in an annular-punctured rabbit.

CCL3/Macrophage Inflammatory Protein-1α Is Dually Involved in Parasite Persistence and Induction of a TNF- and IFNγ-Enriched Inflammatory Milieu in Trypanosoma cruzi-Induced Chronic Cardiomyopathy.

CCL3, a member of the CC-chemokine family, has been associated with macrophage recruitment to heart tissue and parasite control in the acute infection of mouse with Trypanosoma cruzi, the causative agent of Chagas disease. Here, we approached the participation of CCL3 in chronic chagasic cardiomyopathy (CCC), the main clinical form of Chagas disease. We induced CCC in C57BL/6 (ccl3+/+) and CCL3-deficient (ccl3−/−) mice by infection with the Colombian Type I strain. In ccl3+/+ mice, high levels of CCL3 mRNA and protein were detected in the heart tissue during the acute and chronic infection. Survival was not affected by CCL3 deficiency. In comparison with ccl3+/+, chronically infected ccl3−/− mice presented reduced cardiac parasitism and inflammation due to CD8+ cells and macrophages. Leukocytosis was decreased in infected ccl3−/− mice, paralleling the accumulation of CD8+ T cells devoid of activated CCR5+ LFA-1+ cells in the spleen. Further, T. cruzi-infected ccl3−/−mice presented reduced frequency of interferon-gamma (IFNγ)+ cells and numbers of parasite-specific IFNγ-producing cells, while the T. cruzi antigen-specific cytotoxic activity was increased. Stimulation of CCL3-deficient macrophages with IFNγ improved parasite control, in a milieu with reduced nitric oxide (NOx) and tumor necrosis factor (TNF), but similar interleukin-10 (IL-10), concentrations. In comparison with chronically T. cruzi-infected ccl3+/+ counterparts, ccl3−/− mice did not show enlarged heart, loss of left ventricular ejection fraction, QTc prolongation and elevated CK-MB activity. Compared with ccl3+/+, infected ccl3−/− mice showed reduced concentrations of TNF, while IL-10 levels were not affected, in the heart milieu. In spleen of ccl3+/+ NI controls, most of the CD8+ T-cells expressing the CCL3 receptors CCR1 or CCR5 were IL-10+, while in infected mice these cells were mainly TNF+. Lastly, selective blockage of CCR1/CCR5 (Met-RANTES therapy) in chronically infected ccl3+/+ mice reversed pivotal electrical abnormalities (bradycardia, prolonged PR, and QTc interval), in correlation with reduced TNF and, mainly, CCL3 levels in the heart tissue. Therefore, in the chronic T. cruzi infection CCL3 takes part in parasite persistence and contributes to form a CD8+ T-cell and macrophage-enriched cardiac inflammation. Further, increased levels of CCL3 create a scenario with abundant IFNγ and TNF, associated with cardiomyocyte injury, heart dysfunction and QTc prolongation, biomarkers of severity of Chagas' heart disease.

TREATMENT RESPONSE PREDICTION IN PATIENTS WITH RHEUMATOID ARTHRITIS. Review

Relevance. A variety of targeted therapies for rheumatoid arthritis (RA) treatment exist. Therefore, reliable predictors are needed that could be used to accurately predict the efficacy or inefficacy of these therapies in individual patients. This could allow clinicians to improve diagnosis and prognosis, to make the treatment personalized and to reduce healthcare expenses.
 Objectives: to analyze and systemize the predictors of response to treatment in patients with RA.
 Materials and Methods. We analyzed the recently discovered predictors of treatment response in RA patients using papers cited on PubMed, Lilacs, and EMBASE databases from Jan 2005 until Jan 2020. Predictive factors were grouped into four categories: methotrexate (MTX)-treated RA, tumor necrosis factor (TNF)-α inhibitors-treated RA, interleukin (IL)-6 inhibitors-treated RA, and rituximab (RTX)-treated RA.
 Results. Based on the results of several studies, predictors of response to methotrexate were high Disease Activity Score (DAS), concentration of myeloid-related proteins 8/14, high P-glycoprotein levels, low serum calprotectin and leptin levels, baseline serum concentration of tumor necrosis factor (TNF)-α, TNF receptor I, interleukin (IL)-1β, soluble CD163, numbers of CD14+highCD16, vascular cell adhesion molecule, lower expression of hsa-miR-132-3p, hsa-miR-146a-5p, and hsa-miR-155-5p. A positive response to biological therapy was determined by male gender, younger age, lower health assessment questionnaire, erythrocyte sedimentation rate or C-reactive protein, neutrophil-to-lymphocyte ratio, platelet-to-lymphocyte ratio, tender joint count (or swollen joint count) scores, absence of comorbidities, baseline albumin, IL-34, IL-1β, D-dimer, fibrinogen, matrix metalloproteinase 3, DAS 28 and Simplified Disease Activity Index (SDAI). The plasma interferon (IFN) activity and the IFN beta/alpha ratio, IL-1Ra level were predictive in TNF antagonist-treated patients. Predictors of response to IL-6 inhibitors were anti–citrullinated protein antibody (ACPA)+, baseline Sharp/van der Heijde score, myeloid soluble intercellular adhesion molecule 1, serum levels of sIL-6R, IL-8, calprotectin, and lymphoid activation and bone remodeling markers. The prediction of the best response for rituximab was determined to be a combination of IL-33, rheumatoid factor or ACPA, IgG, and also lower number of previous biological therapies. Genetic factors, such as single-nucleotide polymorphisms at gene locus rs10919563, rs11541076, rs12083537, rs11265618, and rs1801274, and rs396991 can also be used to predict a response to treatment.
 Conclusions. One of the leading problems in the development of predictors remains the collection of high-quality and complete information from a large number of patients. For this, it is necessary to develop an digital program for collecting specific data (depending on the specific disease) and developing new algorithms for predicting the response to treatment.

Waterpipe Tobacco Smoke Inhalation Triggers Thrombogenicity, Cardiac Inflammation and Oxidative Stress in Mice: Effects of Flavouring.

The consumption of water-pipe smoking (WPS) has been promoted by the use of flavoured tobacco. However, little is known about the impact of flavouring on the cardiovascular toxicity induced by WPS inhalation. Here, we compared the cardiovascular effects and underlying mechanism of actions of plain (P) (unflavoured) versus apple-flavoured (AF) WPS (30 minutes/day, 5 days/week for 1 month) in mice. Control mice were exposed to air. Both P- and AF-WPS inhalation induced an increase in systolic blood pressure, thrombogenicity and plasma concentration of fibrinogen and von Willebrand factor. In heart homogenates, AF-WPS inhalation caused an increase of 8-isoprostane and a decrease in the levels of reduced glutathione (GSH) and superoxide dismutase (SOD). Nevertheless, P-WPS decreased only the activity of SOD. The concentrations of tumour necrosis factor α and interleukin 1β were increased only in heart homogenates of mice exposed to AF-WPS. Although both P- and AF-WPS increased the concentration of troponin I in heart homogenates and induced DNA damage, the concentration of cleaved caspase 3 was only increased in mice exposed to AF-WPS. Immunohistochemical analysis of the hearts showed that both P- and AF- WPS inhalation decreased the expression of SOD. Moreover, the expression of nuclear factor erythroid-derived 2-like 2 at nuclear level in the heart was higher in both AF-WPS and P-WPS compared with control group, and the effect observed in AF-WPS group was more significant than that seen in P-WPS group. Likewise, the concentration of heme oxygenase-1 was significantly increased in both P-WPS and AF-WPS groups compared with control group, and the effect seen in AF-group was higher than that observed in P-WPS group. In conclusion, our findings showed that both P- and AF-WPS induce thrombogenicity and cardiac injury, and that this toxicity is potentiated by the presence of flavouring.

Abnormal Cannabidiol Affects Production of Pro-Inflammatory Mediators and Astrocyte Wound Closure in Primary Astrocytic-Microglial Cocultures.

Abnormal cannabidiol (abn-CBD) exerts neuroprotective effects in vivo and in vitro. In the present study, we investigated the impact of abn-CBD on the glial production of proinflammatory mediators and scar formation within in vitro models. Primary astrocytic-microglial cocultures and astrocytic cultures from neonatal C57BL/6 mice and CB2 receptor knockout mice were stimulated with lipopolysaccharide (LPS), and the concentrations of tumor necrosis factor α (TNFα), interleukin-6 (IL-6) and nitrite were determined. Furthermore, we performed a live cell microscopy-based scratch-wound assay. After LPS stimulation, TNFα, IL-6 and nitrite production was more strongly increased in cocultures than in isolated astrocytes. Abn-CBD treatment attenuated the LPS-induced production of TNFα and nitrite in cocultures, while IL-6 production remained unaltered. In isolated astrocytes, only LPS-induced TNFα production was reduced by abn-CBD. Similar effects were observed after abn-CBD application in cocultures of CB2 knockout mice. Interestingly, LPS-induced TNFα and nitrite levels were far lower in CB2 knockout cultures compared to wildtypes, while IL-6 levels did not differ. In the scratch-wound assay, treatment with abn-CBD decelerated wound closure when microglial cells were present. Our data shows a differential role of abn-CBD for modulation of glial inflammation and astrocytic scar formation. These findings provide new explanations for mechanisms behind the neuroprotective potential of abn-CBD.

Spatial dynamics of the insulin receptor in living neurons.

Insulin signaling through the insulin receptor has long been studied in classic target organs, such as adipose tissue and skeletal muscle, where one of its effects is to increase glucose uptake. Insulin and insulin receptor are present in many areas of the brain, but the functions of brain insulin signaling outside feeding circuits are not well defined. It has been proposed that hippocampal insulin signaling is important for memory, that brain insulin signaling is deficient in Alzheimer's disease, and that intranasal insulin treatment improves cognition, but the mechanisms remain unclear and do not seem to involve increased glucose uptake by neurons. The molecular behavior of the insulin receptor itself is not well known in living neurons; therefore, we investigated the spatial dynamics of the insulin receptor on somatodendritic membranes of live rat hippocampal neurons in culture. Using single-molecule tracking of quantum dot-tagged insulin receptors and single-particle tracking photoactivation localization microscopy, we show that the insulin receptor is distributed over both dendritic shafts and spines. Using colocalization with synaptic markers, we also show that in contrast to the glutamate receptor subunit glutamate receptor subunit A1, the dynamics of the insulin receptor are not affected by association with excitatory synapses; however, the insulin receptor is immobilized by components of inhibitory synapses. The mobility of the insulin receptor is reduced both by low concentrations of the pro-inflammatory cytokine tumor necrosis factor α and by cholesterol depletion, suggesting an association with sphingolipid-rich membrane domains. On the other hand, the insulin receptor dynamics in hippocampal neurons are not affected by increased excitatory signaling. Finally, using real-time single-event quantification, we find evidence of strong insulin receptor exocytosis on dendritic shafts. Our results suggest an association of the neuronal insulin receptor with specific elements of the dendritic shaft, rather than excitatory synapses.

β-Sitosterol Attenuates High Grain Diet-Induced Inflammatory Stress and Modifies Rumen Fermentation and Microbiota in Sheep.

Simple SummaryOver the years, rumen acidosis is considered as the most common disease in ruminants, which affects animal health and brings huge economic losses of the ruminant industry. Antibiotic have been considered as effective to alleviate the occurrence of ruminal acidosis. However, antibiotic usage in food animals has been absolutely banned by European Union and strictly restricted in other countries. It is necessary to search for safe, effective and healthy additives alternative to antibiotics for ruminants. β-sitosterol is a plant compound known as phytosterol, which has many biological activities. In this study, sheep were feed four levels with β-sitosterol supplementation (0, 0.25, 0.5, and 1.0 g/kg). We concluded that β-sitosterol could attenuate high grain diet-induced inflammatory response and modify ruminal fermentation. These findings provide updated insight for preventing the occurrence of rumen acidosis.β-sitosterol (BSS) is a plant-derived natural bioactive compound, its cellular mechanism of anti-inflammatory activity has been proven recently. Little information is available regarding the application of BSS on ruminants under high grain diet. The objective of this study was to evaluate the effects of dietary BSS supplementation on inflammatory response, ruminal fermentation characteristics and the composition of the ruminal bacterial community under high grain diet. Eight rumen-cannulated Hu sheep (59.7 ± 4.8 kg of initial body weight) were randomly assigned into a replicated 4 × 4 Latin square design trial. Sheep were fed a high grain diet (non-fiber carbohydrate: neutral detergent fiber = 2.03) supplemented either with 0.25 (LBS), 0.5 (MBS), 1.0 (HBS) or without (CON) g BSS /kg dry matter diet. On day 21 of each period, rumen content samples were obtained at 6 h postfeeding, and blood samples were obtained before morning feeding. The data showed that compared with control group, Dietary BSS supplementation decreased serum concentrations of tumor necrosis factor, interleukin (IL)-6, and IL-1β. The ruminal pH and acetate concentration for BSS treatment were improved, while concentration of propionate, butyrate and lactate was decreased. The result of Illumina MiSeq sequencing of 16S rRNA gene revealed that BSS addition can increase the proportion of Prevotella_1, Rikenellaceae_RC9_gut_group, Prevotella_7, and Selenomonas_1, and decrease the proportion of Lachnospiraceae_NK3A20_group. These results indicated that BSS attenuates high grain diet-induced inflammatory response and modifies ruminal fermentation. In addition, the BSS dietary supplementation at the level of 0.5 g/kg is recommended in sheep.