Plasma TNF-α Research Articles

Restoring physiological parameters of the pancreas and kidney through treatment with a polymeric nano-formulation of C-peptide and lisofylline combination in diabetic nephropathy.

Diabetic nephropathy (DN) is a progressive kidney disorder that develops as a complication of diabetes due to long-term exposure to elevated blood glucose levels (BGLs). In this case, an intervention of therapeutic moieties is needed to target the specific elements involved in diabetes to prevent/delay the deterioration of kidney function. Therefore, the present study focused on designing and evaluating a potent nano-formulation of a combination of C-peptide (CPep) and the anti-diabetic drug lisofylline (LSF) to prevent streptozotocin (STZ)-induced DN. As a strategic intervention, an LSF-oleic acid prodrug (LSF-OA) was initially synthesized and further encapsulated in an in-house-synthesized cationic polymer [(mPEG-b-P(CB-{g-DMDP}-co-LA)); mPLM] to prepare polymeric nano-complexes of CPep via electrostatic interaction, possessing a size of 218.6 ± 14.4 nm and zeta potential of +5.2 mV together with stability for 30 days at 25 °C. mPLM-LSF-OA-CPep nanoparticles demonstrated hemocompatibility with RBCs and exhibited potent anti-oxidant activity by reducing nitrite levels, inducing the release of anti-oxidant GSH and protecting metabolically stressed rat kidneys and murine insulinoma cells from apoptosis. In vivo pharmacokinetics depicted an increase in t½ and mean residence time in rats, which further improved the BGL and renal conditions and reduced plasma IL-6 and TNF-α levels in the STZ-induced DN animal model when treated with mPLM-LSF-OA-CPep compared to free LSF and CPep. Moreover, an increase in the plasma insulin level and detection of proliferative marker cells in pancreatic islets suggested the regeneration of β-cells in diabetic animals.

Exploring the efficacy of laser speckle contrast imaging in the stratified diagnosis of rosacea: a quantitative analysis of facial blood flow dynamics across varied regions.

Rosacea has a high incidence, significantly impacts quality of life, and lacks sufficient diagnostic techniques. This study aimed to investigate the feasibility of laser speckle contrast imaging (LSCI) for measuring facial blood perfusion in patients with rosacea and to identify differences in blood flow among various facial regions associated with different rosacea subtypes. From June to December 2023, 45 patients were recruited, with 9 excluded, leaving 36 subjects: 12 with erythematotelangiectatic rosacea (ETR), 12 with papulopustular rosacea (PPR), and 12 healthy controls. The Think View multispectral imaging analyzer assessed inflammation via gray reading values across the full face and five facial areas: forehead, nose, cheeks, and chin. LSCI measured and analyzed blood perfusion in the same areas. Plasma biomarkers interleukin-6 (IL-6), IL-1β, and tumor necrosis factor-α (TNF-α) were tested in different groups. Both ETR and PPR groups showed increased average blood perfusion and facial inflammation intensity by gray values compared to controls, with statistically significant differences. Average blood perfusion of ETR and PPR groups showed increased values in the forehead, cheeks, and nose, compared to controls, and the values in the cheeks were statistically different between ETR and PPR. The facial inflammation intensity of the ETR group showed increased values in the forehead and cheeks, and the PPR group showed increased gray values in the forehead, cheeks, nose, and chin compared to controls, and the values for the cheeks, nose, and chin were statistically significantly different between ETR and PPR. Plasma biomarkers IL-6, IL-1β, and TNF-α were significantly elevated in both ETR and PPR groups compared to controls. LSCI is a valuable, non-invasive tool for assessing blood flow dynamics in rosacea, providing a data foundation for clinical research. Different rosacea subtypes exhibit distinct lesion distribution and blood flow patterns, and both ETR and PPR could affect all facial areas, particularly the cheeks in ETR and the forehead, nose, and chin in PPR.

Restorative effects of (+)-epicatechin in a rodent model of aging induced muscle atrophy: underlying mechanisms.

Sarcopenia is a progressive and generalized age-related skeletal muscle (SkM) disorder characterized by the accelerated loss of muscle mass (atrophy) and function. SkM atrophy is associated with increased incidence of falls, functional decline, frailty and mortality. In its early stage, SkM atrophy is associated with increased pro-inflammatory cytokine levels and proteasome-mediated protein degradation. These processes also link to the activation of atrophy associated factors and signaling pathways for which, there is a lack of approved pharmacotherapies. The objective of this study, was to characterize the capacity of the flavanol (+)-epicatechin (+Epi) to favorably modulate SkM mass and function in a rat model of aging induced sarcopenia and profile candidate mechanisms. Using 23 month old male Sprague-Dawley rats, an 8 weeks oral administration of the +Epi (1 mg per kg per day in water by gavage) was implemented while control rats only received water. SkM strength (grip), treadmill endurance, muscle mass, myofiber area, creatine kinase, lactate dehydrogenase, troponin, α-actin, tumor necrosis factor (TNF)-α and atrophy related endpoints (follistatin, myostatin, NFκB, MuRF 1, atrogin 1) were quantified in plasma and/or gastrocnemius. We also evaluated effects on insulin growth factor (IGF)-1 levels and downstream signaling (AKT/mTORC1). Treatment of aged rats with +Epi, led to significant increases in front paw grip strength, treadmill time and SkM mass vs. controls as well as beneficial changes in makers of myofiber integrity. Treatment significantly reversed adverse changes in plasma and/or SkM TNF-α, IGF-1, atrophy and protein synthesis related endpoints vs. controls. In conclusion, +Epi has the capacity to reverse sarcopenia associated detrimental changes in regulatory pathways leading to improved SkM mass and function. Given these results and its recognized safety and tolerance profile, +Epi warrants consideration for clinical trials.

Expanding the VEXAS diagnostic workup: the role of peripheral blood cytological analysis.

VEXAS syndrome is a newly described autoinflammatory entity characterized by somatic mutations in the UBA1 X-linked gene in hematopoietic progenitor cells. Several studies have demonstrated that the presence of vacuoles in progenitor cells from bone marrow aspirates is a hallmark finding for this syndrome. Therefore, this study aimed to characterize leukocytes from VEXAS patients versus patients with ANCA-associated vasculitis (AAV), familial Mediterranean fever (FMF), and healthy donors (HD) to define a specific cytological pattern that can support VEXAS diagnosis. Twelve VEXAS patients were included in the study. Blood samples from FMF (n = 16), AAV (n = 16) and HDs (n = 20) acted as controls. May-Grünwald Giemsa (MGG) staining was used for studying cellular morphology, including cytoplasm, granules, and vacuoles and to perform a cytogenic evaluation of leucocytes. Plasma IL-1β, IL-1α, TNFα, IL-18 and IL-8 were measured using ELISA assay. The cytological analysis from blood smears confirmed the presence of immature neutrophils in VEXAS patients. We found a greater number of vacuoles in VEXAS patients vs. FMF, AAV and HD. Micronuclei (MNi) and cell death rate were higher in VEXAS patients vs. HD. Cell death correlated with IL-1β and IL-8 levels. MNi were positively associated with IL-8 and IL-1β levels, and with the percentage of immature neutrophils and vacuoles. In conclusion, our findings suggested that cytological test may be supportive for VEXAS diagnosis, despite genetical analysis is mandatory for confirming the disease. Finally, we identified several cytological hallmarks that may distinguish the VEXAS "cytotype" not only from HD but also from other inflammatory diseases.

Malondialdehyde and TNF-α lowering effects of purified gambier (Uncaria gambir Roxb.) in diabetic rats

BackgroundMalondialdehyde (MDA) is one of a dominat marker in oxidative stress condition, and when inflammation occurred tumor necrosis factor- α (TNF-α) played a significant influence in the propagation this process. Purified gambier (Uncaria gambier Roxb.) contained 90% catechin which is proven to have antioxidant activity and may prevent unwanted inflammatory responses during diabetic state. Objective: The objective of this research was to assess how purified gambier affected plasma MDA and TNF- α levels in alloxan-induced diabetic rats. Material and methods: In this study, 35 rats were used. Alloxan 120 mg/kg BW intraperitoneal injection was administered to induce diabetes conditions in rats. All animals were divided into 5 groups, diabetic control group treated with vehicle, positive control group treated with glibenclamide dose 0.45 mg/kg BW), and treatment groups treated with purified gambier dose of 2.5; 5 and 10 mg/kg BW. All animals were treated respectively for 14 days. Plasma MDA and TNF- α levels were measured on day 3, and 14. Results: Two-way ANOVA was applied to analyze all of the data, these findings suggested that purified gambier has antioxidant-related anti-inflammation actions. possesses blood sugar-lowering activity (p<0.05). The plasma MDA and TNF- α level of treatment group were significantly reduced (p<0.05) compared to diabetes control group. Conclusion: These results depicted that at doses of 2.5–10 mg/kg BW, purified gambier has antioxidant-associated anti-inflammation effects when given for 14 days on diabetic rat model by reducing plasma levels MDA and TNF-α.

Plasma TNF-α Elevation in Biologic Naive Rheumatoid Arthritis Patients Belonging to a Population with New Mutations in TLR4 and CYP51A1 genes without Association with Disease-Related Antibodies Levels.

In a system biology-based study, we previously reported that IL-6 and IL6R -specific m-RNA levels were elevated in leukocytes of patients with Rheumatoid arthritis (RA). Here, the association of toll-like receptor4 (TLR4) rs 141534085 and cytochrome P450 family 51 subfamily A member 1(CYP51A1) rs6 with tumor necrosis factor-α (TNF- α), rheumatoid factor (RF)- and Anti- cyclic citrullinated peptide (anti-CCP) antibody -positivity was investigated in almost the same subjects. Forty-six patients and 48 normal subjects were recruited in this study. The blood leucocytes TLR4 rs 141534085 and CYP51A1 rs6 -comprising DNA sequences were amplified by using tetra-primer amplification refractory mutation system polymerase chain reaction (T-ARMS-PCR) technique and the PCR products were checked by Sanger DNA sequencing method. ELISA method was used to determine plasma levels of TNF- α, anti-CCP antibody and RF positivity of plasma was evaluated through a latex agglutination test. The TNF- α level was significantly higher in the patient group than control subjects (p= 0.001). Moreover, we were not able to find any correlation between TNF-α levels and RF as well as anti-CCP antibodies when we used the K2/ Fisher's exact test and Pearson test respectively. Our DNA sequencing data revealed the following new mutations in TLR4 rs141534085 comprising regions: A>T in position 1050, T>A in position 1052, and C>A in position 1054; and for CYP51A1 rs6 encompassing region, the new mutations were; G>A in position 21680, the T nucleotide was inserted in position 21762 and the G nucleotide was inserted in position 21763, G>T in position 21764. The data of this study showed that both TLR4 rs141534085 and CYP51A1 rs6 related DNA regions should be considered as hotspot areas in RA pathogenicity. Moreover, these data indicated that, TNF- α did not alter the production of anti-CCP and RF pathogenic antibodies in patients with long-term RA.

The Potential of UV-B Irradiated Oyster Mushroom (Pleurotus ostreatus) Powder on Plasma Glucose and TNF-α Levels in Type 2 Diabetic Rats

The Potential of UV-B Irradiated Oyster Mushroom (Pleurotus ostreatus) Powder on Plasma Glucose and TNF-α Levels in Type 2 Diabetic Rats

Deleterious effect of chronic high-dose ethanol intake on biomechanical bone properties and periodontal status.

To evaluate the effect of high-graduation chronic ethanol (EtOH) intake on bone and periodontal tissues of rats. Male Wistar rats (250g) were divided into two groups of n = 12 each one. EtOH (5ml of 3g/kg) was administered to the experimental group by gastric gavage twice a day for 20days and the control group received water under the same conditions. The rats were euthanized and used to perform biochemical determination in plasma and gingival tissue, and histological and biomechanical studies in the femur and mandibular tissues. Alcohol increased both TNFα (p < 0.01) and PGE2 (p < 0.05) in plasma and gingiva (p < 0.05) as compared to controls. In addition, EtOH increased the alveolar bone loss as evidenced by the increased distance between the cement enamel junction and the alveolar crest (p < 0.01), the lower % of interradicular bone expressed as bone area/total area (B.Ar/T.Ar, p < 0.05) and the larger periodontal space (p < 0.05), as compared to controls. Likewise, the mandibular microtomographic analysis in alcoholized rats revealed a lower % of interradicular bone volume/total volume (BV/TV, p < 0.05), greater trabecular separation (p < 0.05) and greater % trabecular porosity (p < 0.05) than controls. No biomechanical alteration was observed in lower jaws, while the femur of alcoholized rats presented a decrease in the structural bone properties (p < 0.001), as a systemic consequence of deterioration of the diaphyseal architecture (p < 0.01) without changes in material properties. The consumption of high doses of alcohol produces deleterious effects on periodontal tissues that could be due not only to local but also systemic effects.

TUMOR NECROSIS FACTOR-Α, INTERLUKIN-6, AND VISFATIN LEVELS IN OLDER PATIENTS WITH MALNUTRITION

Abstract Introduction Proinflammatory cytokine is associated with malnutrition status. Tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) were pro-inflammatory cytokines stimulated by nutritional deficiency. Visfatin is an adipokine with a strong correlation with inflammation. The relationships of TNF-α, IL-6 and visfatin are not consistent, and no study have investigated them in the older patients with malnutrition. Methods This prospective study included patients aged ≥ 75 years at the emergency department and all patients underwent Mini-Nutritional Assessment-Short Form (MNA-SF) and blood tests for fasting plasma TNF-α, IL-6 and visfatin levels. Results We enrolled 106 older patients with a mean age of 87.3 years, including 43 (40.5%) patients in at risk of malnutrition group (MNA-SF between 8 to 11), 24 (22.6%) patients in the malnutrition group (MNA-SF &amp;lt; 8), and 39 (36.7%) patients in normal group. Compared to the normal group, both at risk of malnutrition and malnutrition group had significantly lower Mini-Mental State Examination (MMSE). Furthermore, TNF-α was significantly higher in at risk of malnutrition group. In contrast, IL-6 was significantly higher in malnutrition group. Visfatin levels was not correlated to the malnutrition status. Both TNF-α and IL-6 negatively correlated with Barthel index and MMSE. Backward and stepwise multiple logistic regression analyses showed that the independent predictor for both at risk of malnutrition and malnutrition was MMSE. Conclusion The nutrition deficit status was correlated with proinflammatory cytokines, serum TNF-α and IL-6. The independent predictor of both at risk of malnutrition and malnutrition group was MMSE in older patients.

Altered expression of miR-125a and dysregulated cytokines in systemic lupus erythematosus: Unveiling diagnostic and prognostic markers.

Systemic lupus erythematosus (SLE) is a chronic autoimmune disorder impacting multiple organs, influenced by genetic factors, especially those related to the immune system. However, there is a need for new biomarkers in SLE. MicroRNA-125a (miR-125a) levels are decreased in T cells, B cells, and dendritic cells of SLE patients. MiR-125a plays a regulatory role in controlling the levels of tumor necrosis factor-alpha (TNF-α) and interleukin 12 (IL-12), which are crucial pro-inflammatory cytokines in SLE pathogenesis. To assess the levels of miR-125a, IL-12, and TNF-α in SLE patients' plasma, evaluating their diagnostic and prognostic value. The study included 100 healthy individuals, 50 newly diagnosed (ND), and 50 SLE patients undergoing treatment. The patients were monitored for a duration of 24 wk to observe and record instances of relapses. MiR-125a expression was measured using real-time reverse transcription polymerase chain reaction, while ELISA kits were used to assess IL-12 and TNF-α production. The results showed significantly reduced miR-125a expression in SLE patients compared to healthy individuals, with the lowest levels in ND patients. TNF-α and IL-12 expression levels were significantly elevated in SLE patients, especially in the early stages of the disease. Receiver operating characteristic curve analyses, and Cox-Mantel Log-rank tests indicated miR-125a, TNF-α, and IL-12 as proper diagnostic biomarkers for SLE. A negative correlation was found between plasma miR-125a expression and IL-12/TNF-α levels in SLE patients. Decreased miR-125a levels may be involved in the development of SLE, while elevated levels of IL-12 and TNF-α contribute to immune dysregulation. These findings offer new diagnostic and prognostic markers for SLE. Moreover, the negative correlation observed suggests an interaction between miR-125a, TNF-α, and IL-12. Further research is necessary to uncover the underlying mechanisms that govern these relationships.

Plasma neuron specific enolase (NSE), tumour necrosis factor-alpha (TNF-α) and soluble IL-2 receptor alpha (sIL-2Rα) levels in children with developmental delay (DD): Use of combined ROC curves to increase their diagnostic value

BackgroundDevelopmental delay (DD) occurs when children fail to reach developmental milestones in comparison to peers of the same age range. However, there are no valuable biomarkers for the early diagnosis of DD. Since there is no specific marker for screening the disease, we evaluated plasma NSE, TNF-α and sIL2-Rα as potential markers for this purpose. MethodsIn this cross-sectional randomized case-control study, a total of 174 DD patients and 49 matched elderly controls aged between 2 months and 60 months were recruited. A sensitive enzyme-linked immunosorbent assay and an immunoradiometric assay were used to evaluate the levels of plasma IL-1, IL-6, IL-8, IL-10, sIL2-Rα, TNF-α, and NSE. Statistical analyses using t test, χ2, ANOVA, ROC curves and binary logistic regression models were performed. ResultsIn comparison to the control group, the DD group had greater levels of NSE, TNF-α, and sIL2-Rα(p < 0.05). In the binary logistic regression analysis of DD, NSE had an odds ratio (OR) of 1.783 (95 % CI 1.297 to 2.451, p = 0.000), indicating that NSE was an independent risk factor for DD. The plasma TNF-α level was positively correlated with plasma NSE and sIL2-Rα levels in the DD group (r = 0.366 and 0.433, respectively), and the DQ score and plasma sIL2-Rα level in the DD group were positively correlated. The ROC curve revealed that the respective areas under the NSE, TNF-α, and sIL2-Rα ROC curves were 0.9797, 0.9365, and 0.8533, respectively. Moreover, a significant increase in AUC was observed using combined ROC curve analysis. ConclusionsChildren with DD have significantly altered plasma concentrations of sIL2-Rα, NSE, and TNF-α. NSE, TNF-α and sIL2-Rα can be used as DD blood biomarkers. This information may be helpful in early diagnosis and intervention.

Plasma levels of IL-6, IL-10, IL-18, TNFα under the conditions of tobacco intoxication and after treatment with aminophthalhydrazide

In response to damaging factors, a cascade of cytokines is released, triggering the processes of inflammation and fibrogenesis. The purpose of our study was to evaluate the levels of cytokines (IL-6, IL-10, IL-18, TNFα) in blood plasma under conditions of tobacco intoxication and with intramuscular administration of aminophthalhydrazide. The experiment was performed with 40 outbred white male rats. Intact group (n = 10) were in a priming chamber in absence of tobacco smoke. Experimental groups (1, 2, and 3) – n = 30, 10 animals in each group. The first and second groups of animals were exposed to tobacco smoke for one and two months. The third experimental group underwent intramuscular injections with sodium aminodihydrophthalazindione for 3, 7, 14 days. The content of IL-6, IL-10, IL-18, TNFα in blood plasma was determined by enzyme immunoassay technique. In 1st group, the TNFα level in blood plasma increased to 452.14±176.18 pg/mL (in intact animals, 15.23±3.13 pg/mL); in the 2nd group, this index decreased to 9.8; in group 3, these values showed a sharp increase (to 167.44±5.93 pg/mL). The level of IL-6 showed maximal values after 4 months of tobacco intoxication (group 3). The following IL-10 changes were observed: in the 1st group, its amounts increased by 1.2-fold; in the 2nd group a sharp decrease in the indindex was noted (13.7 times); in the 3rd group a decreased content of the cytokine was found (28.8 times) as compared with intact animals. After administration of aminodihydrophthalazinedione for 3 and 7 days, the content of TNFα showed sharp increas (14.8 times and 7.6 times, respectively); after 14 days of treatment, the level of TNFα decreased by 1.3 times compared to intact animals. We noted fluctuations in the levels of pro-inflammatory cytokines (IL-6 and IL-18). The level of IL- 10 in blood plasma had a statistically significant increase (5.3-fold at 3 days after drug administration); 8.2-fold 7 days after administration of the drug, and 23-fold after 14 days of treatment, as compared with the 3rd group. The plasma concentrations of TNFα, IL-6, IL-10, IL-18 showed sufficient changes under the conditions of tobacco intoxication. The immune response is characterized by increased levels of pro-inflammatory cytokines in blood plasma. Upon corrective therapy with sodium aminodihydrophthalazindione, an increase in the antiinflammatory IL-10 cytokine occurs, along with systemic decrease in the TNFα, IL-6, and IL-18 contents.

Modification of Peripheral Blood Flow and Angiogenesis by CO2 Water-Bath Therapy in Diabetic Skeletal Muscle with or without Ischemia.

Previously, it was shown that both blood flow and angiogenesis in the ischemic hind limb of diabetic rats were increased upon CO2 treatment for 4 weeks. In the present study, we have compared the effects of 6 weeks CO2 therapy in diabetic rats with or without peripheral ischemia. Diabetes was induced in rats by a tail vein injection of streptozotocin (65 mg/kg body weight), whereas peripheral ischemia was produced by occluding the femoral artery at 2 weeks of inducing diabetes. Both diabetic and diabetic-ischemic animals were treated with or without CO2 water-bath at 37 °C for 6 weeks (30 min/day; 5 days/week) starting at 2 weeks, after the induction of ischemia. CO2 treatment did not affect heart rate and R-R interval as well as plasma levels of creatine kinase, glucose, cholesterol, triglycerides and high density lipoproteins. Unlike the levels of plasma Ox-LDL, MDA and TNF-α, the levels of NO in diabetic group were increased by CO2 water-bath treatment. On the other hand, the levels of plasma Ox-LDL and MDA were decreased whereas that of NO was increased without any changes in TNF-α level in diabetic-ischemic animals upon CO2 therapy. Treatment of diabetic animals with CO2 increased peak, mean and minimal blood flow by 20, 49 and 43% whereas these values were increased by 53, 26 and 80% in the diabetic-ischemic group by CO2 therapy, respectively. Blood vessel count in diabetic and diabetic-ischemic skeletal muscles was increased by 73 and 136% by CO2 therapy, respectively. These data indicate that peripheral ischemia augmented the increase in blood flow and development of angiogenesis in diabetic skeletal muscle upon CO2 therapy. It is suggested that greater beneficial effects of CO2 therapy in diabetic-ischemic animals in comparison to diabetic group may be a consequence of difference of changes in the redox-sensitive signal transduction mechanisms.

Immunological and biochemical biomarker alterations among SARS-COV-2 patients with varying disease phenotypes in Uganda

Every novel infection requires an assessment of the host response coupled with identification of unique biomarkers for predicting disease pathogenesis, treatment targets and diagnostic utility. Studies have exposed dysregulated inflammatory response induced by the novel severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) as significant predictor or cause of disease severity/prognosis and death. This study evaluated inflammatory biomarkers induced by SARS-CoV-2 in plasma of patients with varying disease phenotypes and healthy controls with prognostic or therapeutic potential. We stratified SARS-CoV-2 plasma samples based on disease status (asymptomatic, mild, severe, and healthy controls), as diagnosed by RT-PCR SARS-CoV-2. We used a solid phase sandwich and competitive Enzyme-Linked Immunosorbent Assay (ELISA) to measure levels of panels of immunological (IFN-γ, TNF-α, IL-6, and IL-10) and biochemical markers (Ferritin, Procalcitonin, C-Reactive Protein, Angiotensin II, Homocysteine, and D-dimer). Biomarker levels were compared across SARS-CoV-2 disease stratification. Plasma IFN-γ, TNF-α, IL-6, and IL-10 levels were significantly (P < 0.05) elevated in the severe SARS-CoV-2 patients as compared to mild, asymptomatic, and healthy controls. Ferritin, Homocysteine, and D-dimer plasma levels were significantly elevated in severe cases over asymptomatic and healthy controls. Plasma C-reactive protein and Angiotensin II levels were significantly (P < 0.05) higher in mild than severe cases and healthy controls. Plasma Procalcitonin levels were significantly higher in asymptomatic than in mild, severe cases and healthy controls. Our study demonstrates the role of host inflammatory biomarkers in modulating the pathogenesis of COVID-19. The study proposes a number of potential biomarkers that could be explored as SARS-CoV-2 treatment targets and possible prognostic predictors for a severe outcome. The comprehensive analysis of prognostic biomarkers may contribute to the evidence-based management of COVID-19 patients.

A novel DYRK1a inhibitor, DYR533, reduces tau pathology and TNF alpha in the 3xTg‐AD and PS19 mouse models

AbstractBackgroundDual specificity tyrosine phosphorylation kinase 1a (DYRK1a) directly phosphorylates tau and amyloid precursor protein and is upregulated in postmortem brain tissue of patients with disorders including Alzheimer’s (AD) and Picks disease (a tauopathy). Our lab has shown that treatment with a DYRK1a inhibitor (DYR219) reduced Aβ plaque deposition and decreased phosphorylated tau at Serine 396 (pTauSer396) in the 3xTg‐AD mouse model. Here, we developed and tested a novel DYRK1a inhibitor (DYR533), which has an increased half‐life and bioavailability, in 3xTg‐AD mice that develop both Aβ and pathological tau, and PS19 mice that develop solely pathological tau.MethodWe dosed eight‐month‐old 3xTg‐AD and four‐month‐old PS19 mice daily with either 1.0‐, 2.5‐, or 5.0‐mg/kg DYR533 or a vehicle dose for two or four months respectively per model. Treatment start age occurred at the onset of neuropathology. Mice underwent rotarod testing to assess motor function and water maze testing to assess spatial learning and memory, with testing starting 2 weeks prior to the end of treatment. Blood and brain tissue were collected for neuropathology assessment. We assessed Aβ 40‐42 in the 3xTg‐AD mice, and pTauSer396 and the pro‐inflammatory cytokine tumor necrosis factor (Tnf)α in the hippocampus of both models. Blood plasma was extracted and analyzed for Tnfα levels.ResultIn both 3xTg‐AD and PS19 mice, DYR533 reduced soluble and insoluble fractions of pTau Ser396 in a dose dependent manner. Notably, DYR533 in 3xTg‐AD mice reduced pTau at threonine 217, which was recently identified as an early marker of AD progression. Soluble cortical Aβ42 levels were significantly reduced in the 1.0 mg/kg and 5.0 mg/kg 3xTg‐AD groups. DYR533 significantly reduced the levels of Tnfα in blood plasma and the hippocampus of both 3xTg‐AD and PS19 mice.ConclusionIn conclusion, the DYRK1a inhibitor, DYR533, reduces pTauSer396, as well as blood plasma and hippocampal levels of Tnf α in both 3xTg‐AD and PS19 models. Ongoing efforts include testing the effects of DYR533 in a rodent model of Down syndrome (Ts65Dn), which harbors three copies of DYRK1a, and completion of 3xTg‐AD and PS19 experiments. Thus far, these results support DYR533 as a potential therapeutic for AD and the tauopathies.

The inflammatory response to birth requires MyD88 and is driven by both mother and offspring

Birth is an inflammatory event for the newborn, characterized by elevations in interleukin (IL)-6, IL-10, and tumor necrosis factor (TNF)-α peripherally and/or centrally, as well as changes in brain microglia. However, the mechanism(s) underlying these responses is unknown. Toll-like receptors (TLRs) play crucial roles in innate immunity and initiate inflammatory cascades upon recognition of endogenous or exogenous antigens. Most TLR signaling depends on the adaptor molecule myeloid differentiation primary response 88 (MyD88). We independently varied MyD88 gene status in mouse dams and their offspring to determine whether the inflammatory response to birth depends on MyD88 signaling and, if so, whether that signaling occurs in the offspring, the mother, or both. We find that the perinatal surges in plasma IL-6 and brain expression of TNF-α depend solely on MyD88 gene status of the offspring, whereas postnatal increases in plasma IL-10 and TNF-α depend on MyD88 in both the pup and dam. Interestingly, MyD88 genotype of the dam primarily drives differences in offspring brain microglial density and has robust effects on developmental neuronal cell death. Milk cytokines were evaluated as a possible source of postnatal maternal influence; although we found high levels of CXCL1/GROα and several other cytokines in ingested post-partum milk, their presence did not require MyD88. Thus, the inflammatory response previously described in the late-term fetus and newborn depends on MyD88 (and, by extension, TLRs), with signaling in both the dam and offspring contributing. Unexpectedly, naturally-occuring neuronal cell death in the newborn is modulated primarily by maternal MyD88 gene status.

Interactions between aspirin and ticagrelor during experimental human endotoxaemia

Abstract Introduction Dual antiplatelet therapy (DAPT) with aspirin 75-100mg once daily (OD) and the P2Y12 inhibitor ticagrelor is a standard treatment for acute coronary syndromes. De-escalating DAPT to ticagrelor monotherapy reduces bleeding risk yet appears to have minimal ischaemic penalty in most groups. The reason for this is unclear. P2Y12 inhibition can inhibit inflammatory processes relevant to atherothrombosis but aspirin may hypothetically counter these via platelet PGE2 inhibition. We studied the regimen-dependent effects of aspirin +/- ticagrelor in a human model of endotoxaemia. Endotoxin activates toll-like receptor 4, a key damage receptor in atherogenesis. Methods 72 healthy volunteers were randomised to receive either no aspirin or aspirin 20mg twice daily (BD), 75mg OD or 300mg OD for 10 days +/- a loading dose of ticagrelor on the last day prior to receiving intravenous endotoxin administration (2ng/kg). Blood was collected at 9 timepoints from 1h pre to 6h post endotoxin. Key cytokines and prostanoids in the blood/urine were measured by ELISA, platelet function by light transmittance aggregometry and platelet-leukocyte interaction by flow cytometry. Bleeding time was measured pre and 3h post endotoxin. After &amp;gt;5 weeks, in a 2nd period they received the same dose of aspirin as the 1st but received ticagrelor if they had not already, and vice versa, undergoing a 2nd endotoxin challenge at the end of the period. The primary endpoint was plasma tumour necrosis factor (TNF)-α. Results 72 participants were randomised (median 22.5yrs [range 18-59], 6F:66M). A total of 128 endotoxin challenges were performed. Endotoxaemia was associated with flu-like symptoms, pyrexia, tachycardia and relative hypotension but was safe. In those not receiving aspirin, plasma TNF-α was significantly lower when receiving ticagrelor vs. no ticagrelor, but in those receiving any dose of aspirin the significant effect of ticagrelor was abolished (Fig 1). All doses of aspirin fully suppressed arachidonic acid-induced PA with or without ticagrelor despite the thromboinflammatory stimulus of endotoxaemia (Fig 2A), and significantly augmented the inhibitory effect of ticagrelor on collagen-induced PA (2B). Aspirin 75mg OD and 300mg OD, but not 20mg BD, significantly prolonged bleeding time from baseline (2C). Ticagrelor significantly prolonged bleeding time in combination with any dose of aspirin or no aspirin. Aspirin dose-dependently inhibited PGE2 release. Conclusions All doses of aspirin tested prevented the ability of ticagrelor to significantly inhibit cytokine release during experimental endotoxaemia. Aspirin 20mg BD may offer improved bleeding risk with maintained antiplatelet effect compared to standard regimens. Where ticagrelor monotherapy is an option, dropping aspirin may allow ticagrelor to exert greater beneficial effect on inflammatory pathways relevant to atherothrombosis, potentially reducing ischaemic risk whilst also reducing bleeding risk.Figure 1Figure 2

245 Fish Oil and Dexamethasone Administration Each Partially Improve Indicators of Glucose Metabolism and Fat Homeostasis By Reducing Systemic Inflammation in Heat-Stressed Feedlot Wethers

Abstract Heat stress is costly for livestock production due to its impact on metabolism, which negatively affects health and growth. Previous pair-fed studies demonstrated that heat stress-induced inflammation is associated with disruptions in metabolism and growth. Therefore, the objective of this study was to determine whether targeting systemic inflammation in heat-stressed wethers improves metabolic and stress indicators. Feedlot wethers were randomly assigned to 30-d heat stress (40°C; THI = 85) or were pair-fed under thermoneutral conditions (controls; 25°C; n = 11). Heat-stressed wethers were administered oral fish oil capsules twice daily (HeatStress+FO; 2,400 mg/d; n = 11), injected with dexamethasone every 3 d (HeatStress+DEX; 0.15 mg/kg, i.m.; n = 11), or received no intervention (HeatStress; n = 11). Plasma hormones and lipids were assessed over the 30-d period, and soleus muscle and adipose tissue were collected at necropsy for ex vivo glucose metabolism studies and non-esterified fatty acid (NEFA) mobilization analyses, respectively. Plasma TNFα was up to 3-fold greater (P &amp;lt; 0.05) in HeatStress wethers than controls throughout the study. By the end of the study, plasma TNFα in HeatStress+DEX and HeatStress+FO was reduced (P &amp;lt; 0.05) by 38% and 47%, respectively, compared with HeatStress and did not differ from controls. Plasma insulin did not differ among groups early in the study but by d 30 were increased (P &amp;lt; 0.05) in HeatStress and HeatStress+DEX compared with controls and HeatStress+FO. Blood glucose-to-insulin ratios were less (P &amp;lt; 0.05) for HeatStress wethers than controls throughout the study and were intermediate for HeatStress+DEX and HeatStress+FO. Plasma cortisol did not differ among groups and was less (P &amp;lt; 0.05) at d 30 than d -3 for all weathers. Plasma eicosapentaenoic acid and triglycerides were 15% to 22% less (P &amp;lt; 0.05) for HeatStress and HeatStress+DEX but not HeatStress+FO than for controls throughout the study. High density lipoprotein-bound cholesterol was 8% and 22% less (P &amp;lt; 0.05), respectively, for HeatStress and HeatStress+DEX wethers but not HeatStress+FO wethers compared with controls. Ex vivo basal and insulin-stimulated glucose oxidation rates were reduced (P &amp;lt; 0.05) by 32% in muscle from HeatStress wethers but not HeatStress+DEX, or HeatStress+FO wethers compared with muscle from controls. Glucose uptake rates did not differ among groups. Mobilization of NEFA was reduced (P &amp;lt; 0.05) by 19% and 37%, respectively, in fat from HeatStress wethers and HeatStress+DEX wethers compared with controls and was intermediate in fat from HeatStress+FO wethers. These data indicate that supplementation of DEX and FO partially improve muscle-specific glucose metabolism as well as lipid homeostasis and stress indicators, which provides a fundamental basis for heat stress interventions that utilize anti-inflammatory supplements.

240 Prolonged Maternofetal Inflammation in the Pregnant Ewe at Mid-Gestation Increases Subsequent Fetal Inflammatory Tone and Disrupts Skeletal Muscle Growth and Glucose Metabolism Near Term

Abstract Chronic maternofetal inflammation in late gestation (i.e., peak fetal growth) causes fetal intrauterine growth restriction (IUGR) in sheep, which was associated with deficits in growth, body composition, and metabolic function before and after birth. However, the impact of sustained maternofetal inflammation during mid-gestation (i.e., peak placental growth and development) has not been explored. Therefore, our objective was to determine the effects of sustained maternofetal inflammation during this critical window on subsequent fetal inflammatory tone, metabolism, and myoblast (i.e., muscle stem cell) function. Pregnant ewes were injected every 3rd day from the 50th to 65th d of gestational age (dGA; term = 150 dGA) with saline (control; n = 12) or bacterial lipopolysaccharide endotoxin to induce maternofetal inflammation and IUGR (MI-IUGR; n = 12). Fetuses were surgically catheterized on dGA 118 to collect blood samples, and a hyperinsulinemic-euglycemic clamp (HEC) was performed on dGA 123. Fetuses were necropsied on dGA 125 to collect semitendinosus muscle samples. Fetal plasma TNFα concentrations tended to be greater (P = 0.07) for MI-IUGR fetuses (440 ± 45 pg/m l) than for controls (348 ± 24 pg/mL). Fetal blood O2 partial pressures were 12% less (P &amp;lt; 0.05) and maternofetal O2 gradients tended to be 11% greater (P = 0.08) for MI-IUGR fetuses than for controls. Hindlimb-specific glucose uptake rates did not differ between fetal groups, but hindlimb-specific glucose oxidation rates were reduced (P &amp;lt; 0.05) by 15% under basal conditions and by 27% under hyperinsulinemic conditions for MI-IUGR fetuses compared with controls. Semitendinosus glycogen content and β2 adrenoceptor content did not differ between fetal groups, but semitendinosus content of the TNFα receptor, TNFR1, was 41% greater (P &amp;lt; 0.05) for MI-IUGR fetuses than for controls. Immunohistochemistry revealed 18% fewer (P &amp;lt; 0.05) proliferating myoblasts (pax7+/Ki67+) and 38% fewer (P &amp;lt; 0.05) differentiated myoblasts (myogenin+) in semitendinosus from MI-IUGR fetuses compared with controls, which coincided with 13% smaller (P &amp;lt; 0.05) muscle fibers. From these findings, we conclude that sustained maternofetal inflammation during peak placental development at mid-gestation resulted in enhanced fetal inflammatory tone near term, which in turn impaired myoblast function, muscle fiber hypertrophy, and muscle glucose metabolism.

241 Increased Inflammatory Tone and Impaired Skeletal Muscle Growth in Intrauterine Growth-Restricted Neonatal Lambs Is Improved By Daily Supplementation of ω-3 Polyunsaturated Fatty Acid Ca2+ Salts

Abstract Low birthweight due to fetal intrauterine growth restriction (IUGR) often results from maternofetal heat stress. IUGR fetuses exhibit enhanced inflammatory tone, which persists after birth and impacts body composition, skeletal muscle growth, and metabolic function. Therefore, our objective was to determine whether daily oral supplementation of Ca2+ salts of ω-3 polyunsaturated fatty acids (ω-3 PUFA) to IUGR-born neonatal lambs improved inflammatory tone, body composition, and growth metrics. Pregnant ewes were exposed to heat stress (40°C, 35% humidity, THI = 85) from d 40 to 95 of gestation to induce IUGR. Lambs were weaned at birth and hand-reared on milk replacer. Daily oral boluses of 0.5 mL molasses containing 0 or 0.42 g/kg bodyweight of ω-3 PUFA (Virtus Nutrition) were administered. Femoral catheters were placed at d 24 and lambs were necropsied at d 30. At birth, IUGR lambs (n = 8; 3.12 ± 0.34 kg) and IUGR+ω-3 lambs (n = 7; 3.67 ± 0.32 kg) were lighter (P &amp;lt; 0.05) than controls (n = 12; 4.28 ± 0.15 kg). Plasma TNFα concentrations were greater (P &amp;lt; 0.05) at birth for IUGR (557 ± 121 pg/mL) and IUGR+ω-3 lambs (437 ± 82 pg/mL) than for controls (213 ± 78 pg/mL). Plasma TNFα concentrations remained increased (P &amp;lt; 0.05) for IUGR lambs but not for IUGR+ω-3 compared with controls at 2, 3, and 4 wk of age. Adjusted 30-d bodyweights were lighter (P &amp;lt; 0.05) for IUGR lambs (9.83 ± 0.6 kg) but not for IUGR+ω-3 lambs (11.59 ± 0.4 kg) than for controls (12.35 ± 0.2 kg). Heart and brain weights were 17% and 6% lighter (P &amp;lt; 0.05), respectively, for IUGR lambs than for controls. Brain weight deficits were resolved in IUGR+ω3 lambs, but heart weights did not differ between IUGR and IUGR+ω-3 IUGR lambs. Crown circumference/ bodyweight, abdominal circumference/ bodyweight, and cannon bone length/ bodyweight (indicators of asymmetric body composition) were 11% to 15% greater (P &amp;lt; 0.05) for IUGR lambs than for IUGR+ω-3 lambs or controls. At necropsy, IUGR lambs had lighter (P &amp;lt; 0.05) whole hindlimbs (29%) and flexor digitorum superficialis (24%), soleus (36%), biceps femoris (25%), semitendinosus (24%), gastrocnemius (29%), and longissimus dorsi (26%) muscles compared with controls. Weight deficits were partially or fully resolved in IUGR+ω-3 lambs for whole hindlimbs and for all muscles but the soleus, which did not differ from IUGR lambs. These findings demonstrate that daily oral supplementation of Ca2+ salts of ω-3 PUFA reduced the enhanced inflammatory tone observed in IUGR-born offspring, which coincided with improved skeletal muscle growth and indicators of symmetrical body composition.