Inflammatory Markers Myeloperoxidase Research Articles

Antiparkinson potential of khellin on rotenone-induced Parkinson's disease in a zebrafish model: targeting MAO, inflammatory, and oxidative stress markers with molecular docking, MD simulations, and histopathology evidence

In this study, the antiparkinson effect of khellin (KL) on rotenone-induced Parkinson's disease (PD) was examined in zebrafish. Initially, In silico evaluations, such as drug likeness and ADME/T analysis, confirmed the pharmacological viability of KL. Molecular docking and molecular dynamics (MD) analysis revealed stable binding interactions between KL and monamine oxidase B (MAO-B). Molecular docking results for KL and pioglitazone (CCl) revealed binding energies of −6.5 and −10.4 kcal/mol, respectively. Later, molecular dynamics (MD) studies were performed to assess the stability of these complexes, which yielded binding energies of −36.04 ± 55.21 and −56.2 ± 80.63 kJ/mol for KL and CCl, respectively. These results suggest that KL exhibits considerable binding affinity for MAO-B. In In vitro studies, according to the DPPH free radical scavenging assay, KL exhibited significant antioxidant effects, indicating that it can promote redox balance with an IC50 value of 22.68 ± 0.5 μg/ml. In vivo studies and evaluation of locomotor activity, social interaction, histopathology and biochemical parameters were conducted in KL-treated zebrafish to measure SOD and GSH antioxidant activity, the oxidative stress marker malondialdehyde (MDA), the inflammatory marker myeloperoxidase (MPO) and MAO-B. However, while the locomotor and social interaction abilities of the rotenone-treated zebrafish were significantly reduced, KL treatment significantly improved locomotor activity (p < 0.001) and social interaction (p < 0.001). KL alleviated PD symptoms, as indicated by significant increases in SOD (p < 0.01), GSH (p < 0.001), MDA (p < 0.001), MAO-B (p < 0.001) and MPO (p < 0.001) in rotenone-induced PD fish (p<0.001) significantly reduced activities. Histopathological studies revealed that rotenone-induced brain hyperintensity and abnormal cellularity of the periventricular gray matter in the optic tectum were significantly reduced by KL treatment. This study provides a strong basis for developing KL as a new candidate for the treatment of Parkinson's disease, with the prospect of improved safety profiles and efficacy.

Whole intestinal microbiota transplantation is more effective than fecal microbiota transplantation in reducing the susceptibility of DSS-induced germ-free mice colitis.

Fecal microbiota transplantation (FMT) is an emerging and effective therapy for the treatment of inflammatory bowel disease (IBD). Previous studies have reported that compared with FMT, whole intestinal microbiota transplantation (WIMT) can more precisely replicate the community structure and reduce the inflammatory response of the host. However, it remains unclear whether WIMT is more effective in alleviating IBD. To examine the efficacy of WIMT and FMT in the intervention of IBD, GF (Germ-free) BALB/c mice were pre-colonized with whole intestinal microbiota or fecal microbiota before being treated with dextran sodium sulfate (DSS). As expected, the symptoms of colitis were alleviated by both WIMT and FMT, as demonstrated by the prevention of body weight loss and decreased the Disease activity index and histological scores in mice. However, WIMT's anti-inflammatory effect was superior to that of FMT. In addition, the inflammatory markers myeloperoxidase (MPO) and eosinophil peroxidase were dramatically downregulated by WIMT and FMT. Furthermore, the use of two different types of donors facilitated the regulation of cytokine homeostasis in colitis mice; the level of the pro-inflammatory cytokine IL-1β in the WIMT group was significantly lower than that in the FMT group, while the level of the anti-inflammatory factor IL-10 was significantly higher than that in the FMT group. Both groups showed enhanced expression of occludin to protect the intestinal barrier in comparison with the DSS group, and the WIMT group demonstrated considerably increased levels of ZO-1. The sequencing results showed that the WIMT group was highly enriched in Bifidobacterium, whereas the FMT group was significantly enriched in Lactobacillus and Ochrobactrum. Correlation analysis revealed that Bifidobacterium was negatively correlated with TNF-α, whereas Ochrobactrum was positively correlated with MPO and negatively correlated with IL-10, which might be related to different efficacies. Functional prediction using PICRUSt2 revealed that the FMT group was considerably enriched in the L-arginine biosynthesis I and L-arginine biosynthesis IV pathway, whereas the WIMT group was enriched in the L-lysine fermentation to acetate and butanoate pathway. In conclusion, the symptoms of colitis were subsided to varying degrees by the two different types of donors, with the WIMT group being more effective than the FMT group. This study provides new information on clinical interventions for IBD.

Human Fecal Microbiota Transplantation Reduces the Susceptibility to Dextran Sulfate Sodium-Induced Germ-Free Mouse Colitis.

In clinical practice, fecal microbiota transplantation (FMT) has been used to treat inflammatory bowel disease (IBD), and has shown certain effects. However, the selection of FMT donors and the mechanism underlying the effect of FMT intervention in IBD require further exploration. In this study, dextran sodium sulfate (DSS)-induced colitis mice were used to determine the differences in the protection of colitis symptoms, inflammation, and intestinal barrier, by FMT from two donors. Intriguingly, pre-administration of healthy bacterial fluid significantly relieved the symptoms of colitis compared to the ulcerative colitis (UC) bacteria. In addition, healthy donor (HD) bacteria significantly reduced the levels of inflammatory markers Myeloperoxidase (MPO) and Eosinophil peroxidase (EPO), and various pro-inflammatory factors, in colitis mice, and increased the secretion of the anti-inflammatory factor IL-10. Metagenomic sequencing indicated higher species diversity and higher abundance of anti-inflammatory bacteria in the HD intervention group, including Alistipes putredinis, Akkermansia muciniphila, Bifidobacterium adolescentis, short-chain fatty acids (SCFAs)-producing bacterium Christensenella minuta, and secondary bile acids (SBAs)-producing bacterium Clostridium leptum. In the UC intervention group, the SCFA-producing bacterium Bacteroides stercoris, IBD-related bacterium Ruminococcus gnavus, Enterococcus faecalis, and the conditional pathogen Bacteroides caccae, were more abundant. Metabolomics analysis showed that the two types of FMT significantly modulated the metabolism of DSS-induced mice. Moreover, compared with the UC intervention group, indoleacetic acid and unsaturated fatty acids (DHA, DPA, and EPA) with anti-inflammatory effects were significantly enriched in the HD intervention group. In summary, these results indicate that FMT can alleviate the symptoms of colitis, and the effect of HD intervention is better than that of UC intervention. This study offers new insights into the mechanisms of FMT clinical intervention in IBD.

405 Effect of Subacute Ruminal Acidosis (SARA) and Saccharomyces cerevisiae fermentation products on inflammatory responses of dairy cows

Abstract Feeding high-grain diets to dairy cows increases concentrations of acute phase proteins in peripheral blood. This inflammatory response can be reduced by supplementation with Saccharomyces cerevisiae fermentation products (SCFP). It is not clear which cytokines drive this inflammatory response, which organs are inflamed, and how these processes are affected by SCFP. The objectives of this study were to investigate if grain-induced SARA increases the concentrations of the cytokines IL-1β in peripheral blood, and of the inflammatory marker myeloperoxidase (MPO) in rumen papillae of lactating dairy cows, and if these concentrations are affected by SCFP. Thirty-two lactating Holstein dairy cows were randomly assigned to four treatment groups (n = 8) that received a TMR (34.9 %DM NDF, 18.6 %DM starch) supplemented with 1) 140 g/d of ground corn (Control), 2) 126 g/d corn and 14 g/d of Diamond V Original XPCTM (XPC), 3) 121 g/d corn and 19 g/d Diamond V NutriTek® (NTL), and 4) 102 g/d corn and 38 g/d of Diamond V NutriTek® (NTH). SARA challenges were conducted during wk 5 (SARA1) and 8 (SARA2) of lactation by replacing 20% of the base TMR with pellets containing 50% barley and 50% wheat. Blood samples were taken weekly between wk 4 (preSARA1) and wk 9 (postSARA2) for the analysis of IL-1β. Rumen papillae samples were taken during wk 3 (preSARA1) and wk 9 (postSARA2) for the measurement of MPO. SCFP treatment did not affect the concentrations of IL-1β, but the SARA challenges increased this concentration moderately from 9.0 to 12.3 pg/mL (P &amp;lt; 0.05). The concentration of MPO did not differ between preSARA1 and postSARA2, and these concentrations were not affected by SCFP. Results suggest that IL-1β may drive the acute phase response during the SARA challenges, and that these challenges did not cause inflammation of rumen papillae.

The mechanisms for activation of a vascular platelet component of hemostasis in the stroke recovery period and the ways of their correction

Objective : to assess changes in the vascular platelet component of hemostasis in patients in different recovery periods of ischemic stroke (IS). Patients and methods. The investigation enrolled 73 patients with prior IS. According to the remoteness of stroke, the patients were divided into two groups: 1) 41 patients with a stroke remoteness of 1 to 6 months (an early recovery period); 2) 32 patients with that of 7 to 12 months (a late recovery period). In addition, a group of patients with recurrent stroke was identified to evaluate the efficiency of secondary prevention. A control group consisted of 30 healthy volunteers. All the patients took acetylsalicylic acid (ASA). Medical history data and laboratory and instrumental findings were analyzed. To assess the vascular platelet component of hemostasis, platelet aggregation with inductors was studied applying optical aggregometry; enzyme immunoassay was used to estimate the concentrations of the inflammatory marker myeloperoxidase and the endothelial dysfunction (ED) markers sICAM-1 and sE-selectin. Results and discussion. The patients were found to have enhanced platelet aggregation with inductors; moreover, this was more pronounced in the patients with recurrent stroke than in those with new-onset IS. This suggests that the vascular platelet component of hemostasis contributes to the progression of the disease and its unfavorable course and necessitates the monitoring of these parameters to choose optimal secondary prevention methods, such as replacement of ASA with clopidogrel or use of its combination with dipyridamol. The activity of chronic immune inflammation processes and ED was ascertained to be enhanced, which can contribute to the aggravation of cerebrovascular insufficiency and to the development of acute cerebrovascular accident. Conclusion. In the recovery period of stroke, the follow-up monitoring of hemostatic parameters and immune inflammatory and ED markers is of importance in evaluating the efficiency of secondary prevention. For the successful prevention of recurrent strokes, it is necessary to prescribe adequate antiplatelet therapy, as well as drugs that have anti-inflammatory properties, affect intercellular interaction processes (ASA, clopidogrel, and statins), and suppress oxidative stress associated with endothelial inflammation (alpha-lipoic acid and succinic acid preparations).

Inflammation, angiogenesis and fibrogenesis are differentially modulated by distinct domains of the snake venom metalloproteinase jararhagin

Jararhagin, a metalloprotease from Bothrops jararaca snake venom, is a toxin containing the metalloproteinase, disintegrin-like and cysteine-rich domains; it causes acute inflammation and damage to vascular tissue. However, the actions of these domains on key components of chronic inflammation have not been determined. Our aim was to investigate the effects of jararhagin (Jar), jararhagin-C (Jar-C) and o-phenantrolin-treated jararhagin (Jar-Phe), on inflammatory response, blood vessel formation and extracellular matrix deposition in the murine sponge model. The polyether-polyurethane sponge matrix was implanted into Balb/c mice and injected daily with Jar (400 ng), Jar-Phe (400 ng), Jar-C (200 ng) or saline (control). Nine days after implantation, the sponge discs were removed and processed. In the Jar-treated implants, some of inflammatory markers (N-acetyl-β-d-glucosaminidase activity, CCL2 and TNF-α) and TGF-β1 levels were higher compared with the control group. In the Jar-C group, the inflammatory markers myeloperoxidase activity and CXCL1 were higher compared with the control. In this group, VEGF levels and collagen deposition were also higher. Jar-Phe treatment was able to inhibit the activity and/or production of MPO, CXCL1, CCL2 and TGF-β. The differential effects of these proteins in modulating the main components of fibrovascular tissue may be exploited in the management fibroproliferative diseases.

Allantoin as an independent marker associated with carotid intima-media thickness in subclinical atherosclerosis.

Allantoin is the main product of uric acid oxidation and was found to be augmented in atherosclerotic plaque in human autopsy and in animal models of atherosclerosis. Uric acid is abundant in human plasma and is prone to oxidation in inflammatory conditions such as atherosclerosis. In this study, we found a significant increase in plasma uric acid (P=0.002) and allantoin (P=0.025) in participants of the Brazilian Longitudinal Study of Adult Health (ELSA-Brasil) that presented common carotid intima-media thickness (c-IMT) within the 75th percentile (c-IMT≥P75). Multiple linear regression showed an association of c-IMT with uric acid (β=0.0004, P=0.014) and allantoin (β=0.018, P=0.008). This association was independent of age, the traditional risk factor LDL/HDL ratio, and non-traditional risk factors: pulse pressure, neck circumference, and the inflammatory marker myeloperoxidase. The independent and strong association of allantoin with c-IMT shows that it might be a useful marker, along with other traditional risk factors, to evaluate an early stage of atherosclerosis.

Impact of Obesity on Orthodontic Tooth Movement in Adolescents: A Prospective Clinical Cohort Study

Obesity is a widespread chronic inflammatory disorder characterized by an increased overall disease burden and significant association with periodontitis. The aim of this prospective clinical cohort study was to investigate the effect of obesity on orthodontic tooth movement. Fifty-five adolescent patients (27 males, 28 females) with a mean (SD) age of 15.1 (1.7) years and mean (SD) body mass index (BMI) of 30.2 (3.5) kg/m2 in obese and 19.4 (2.2) kg/m2 in normal-weight groups were followed from start of treatment to completion of tooth alignment with fixed orthodontic appliances. Primary outcome was time taken to complete tooth alignment, while secondary outcomes included rate of tooth movement and change in clinical parameters (plaque/gingival indices, unstimulated whole-mouth salivary flow rate, gingival crevicular fluid biomarkers). Data collection took place at baseline (start of treatment: appliance placement), 1 h and 1 wk following appliance placement, and completion of alignment. Results were analyzed by descriptive statistics followed by generalized estimating equation regression modeling. There were no significant differences between groups in time taken to achieve tooth alignment (mean [SD] 158.7 [75.3] d; P = 0.486). However, at 1 wk, initial tooth displacement was significantly increased in the obese group (P < 0.001), and after adjusting for confounders, obese patients had a significantly higher rate of tooth movement compared with normal-weight patients (+0.017 mm/d; 95% confidence interval, 0.008–0.025; P < 0.001) over the period of alignment. Explorative analyses indicated that levels of the adipokines leptin and resistin, the inflammatory marker myeloperoxidase (MPO), and the cytokine receptor for nuclear factor kappa-B ligand (RANKL) were significantly different between obese- and normal-weight patients and associated with observed rates of tooth movement. This represents the first prospective data demonstrating a different response in obese patients compared with normal-weight patients during early orthodontic treatment. These differences in the response of periodontal tissues to orthodontic force in the presence of obesity have potential short- and long-term clinical implications.

Systematic investigation of ethanolic extract from Leea macrophylla: Implications in wound healing

Ethnopharmacological relevanceLeea macrophylla Roxb. ex Hornem. (Leeaceae) commonly known as Hastikarnapalasa is mainly distributed throughout the tropical parts of India. Traditionally, the plant is found to be effective against guinea worm, ringworm and is applied to sores and wounds. Aim of the studyThe present study aims to validate traditional wound healing claim of Leea macrophylla scientifically. Material and methodsBox–Behnken design (BBD) was used to optimize the extraction process. The optimized root tuber extract of Leea macrophylla was standardized with chlorogenic acid by HPLC for the first time. Both oral and topical routes were selected as administrative means for the wound healing study using excision and incision wound model. For topical treatment bioadhesive gel was formulated and characterized for mechanical and physical characteristics by texture profile analysis (TPA) and scanning electron microscopy (SEM). The effect on wound healing was also assessed by evaluating antioxidant enzymes viz. glutathione (GSH), superoxide dismutase (SOD) and catalase (CAT), free radicals lipid peroxidation (LPO) and nitric oxide (NO), inflammatory marker myeloperoxidase (MPO), collagen markers hydroxyproline, hexosamine and hexuronic acid along with the histopathological examination. Furthermore, the effect on the level of the proinflammatory cytokines interleukin-1β (IL-1β), interleukin -6 (IL-6), tumor necrosis factor-α (TNF-α) and growth factor, vascular endothelial growth factor (VEGF) were determined. The expression of cell proliferation nuclear marker Ki-67 was also analyzed by Western blot analysis. ResultsWith mesh openings Sieve no. 20, semi polar nature of solvent (92.5:7.5 ethanol-water blend) and extraction time of 18h, substantially greater extraction efficiency (29%) and phenolic yield (181.54mg/g) were obtained. The content of chlorogenic acid in ethanol extracts of Leea macrophylla was obtained as 9.01% w/w. In incision model, oral treatment with 500mg/kg ethanolic extract increased wound breaking strength by 23.41% while bioadhesive gel (5% w/v) showed a higher increase of 44.68%. Topical application produced complete wound contraction in 20 days against 22 days taken by oral treatment. Topical treatment also produced a significant (p<0.05) increase in antioxidants glutathione, superoxide dismutase and catalase whereas the level of enzymes lipid peroxidation and nitric oxide and inflammatory markers myeloperoxidase were reduced. Further advantageous effects were reflected by significantly (p<0.05) increased levels of hydroxyproline, hexosamine and hexuronic acid. Favorable effects on the level of proinflammatory cytokines interleukin-1β, interleukin-6, tumor necrosis factor – α and growth factor, vascular endothelial growth factor were also observed. The wound healing potential of Leea macrophylla was further supported by its ability to promote cell proliferation during wound healing as demonstrated by Western blot analysis of proliferation marker Ki-67. ConclusionThe study justified traditional use of Leea macrophylla in wound healing and demonstrated that the bioadhesive gel of ethanolic extract produced faster and more significant healing as compared to oral treatment.

Milk Intolerance, Beta-Casein and Lactose.

True lactose intolerance (symptoms stemming from lactose malabsorption) is less common than is widely perceived, and should be viewed as just one potential cause of cows’ milk intolerance. There is increasing evidence that A1 beta-casein, a protein produced by a major proportion of European-origin cattle but not purebred Asian or African cattle, is also associated with cows’ milk intolerance. In humans, digestion of bovine A1 beta-casein, but not the alternative A2 beta-casein, releases beta-casomorphin-7, which activates μ-opioid receptors expressed throughout the gastrointestinal tract and body. Studies in rodents show that milk containing A1 beta-casein significantly increases gastrointestinal transit time, production of dipeptidyl peptidase-4 and the inflammatory marker myeloperoxidase compared with milk containing A2 beta-casein. Co-administration of the opioid receptor antagonist naloxone blocks the myeloperoxidase and gastrointestinal motility effects, indicating opioid signaling pathway involvement. In humans, a double-blind, randomized cross-over study showed that participants consuming A1 beta-casein type cows’ milk experienced statistically significantly higher Bristol stool values compared with those receiving A2 beta-casein milk. Additionally, a statistically significant positive association between abdominal pain and stool consistency was observed when participants consumed the A1 but not the A2 diet. Further studies of the role of A1 beta-casein in milk intolerance are needed.

Orally administrated Lactobacillus pentosus var. plantarum C29 ameliorates age-dependent colitis by inhibiting the nuclear factor-kappa B signaling pathway via the regulation of lipopolysaccharide production by gut microbiota.

To evaluate the anti-inflammaging effect of lactic acid bacteria (LAB) on age-dependent inflammation, we first screened and selected a tumor necrosis factor (TNF)-α and reactive oxygen species (ROS)-inhibitory LAB, Lactobacillus pentosus var. plantarum C29, among the LABs isolated from fermented vegetables using LPS-stimulated mouse peritoneal macrophages. Oral administration of C29 (2 × 109 CFU/rat) for 8 weeks in aged Fischer 344 rats (age, 16 months) inhibited the expression of the inflammatory markers myeloperoxidase, inducible nitric oxide (NO) synthase, cyclooxygenase-2, pro-inflammatory cytokines tumor necrosis factor (TNF)-α and IL-6 and the activation of nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB), activator protein 1 (AP1), and mitogen-activated protein kinases (MAPKs). Treatment with C29 induced the expression of tight junction proteins ZO-1, occludin, and claudin-1, and reduced intestinal microbial LPS and plasmatic LPS levels and ROS, as well as the Firmicutes to Bacteroidetes ratio, which is significantly higher in aged rats than in young rats. C29 treatment also reduced plasmatic reactive oxygen species, malondialdehyde, C-reactive protein, and TNF-α, and suppressed expression of senescence markers p16 and p53 in the colon of the aged rats, but increased SIRT 1 expression. Based on these findings, we concluded that C29 treatment may suppress aging-dependent colitis by inhibiting NF-κB, AP1, and MAPK activation via the inhibition of gut microbiota LPS production and the induction of tight junction protein expression.

Altered vasoactive intestinal peptides expression in irritable bowel syndrome patients and rats with trinitrobenzene sulfonic acid-induced colitis.

To investigate the vasoactive intestinal peptides (VIP) expression in irritable bowel syndrome (IBS) and trinitrobenzene sulfonic acid (TNBS) induced colitis. The VIP gene expression and protein plasma levels were measured in adult participants (45.8% male) who met Rome III criteria for IBS for longer than 6 mo and in a rat model of colitis as induced by TNBS. Plasma and colons were collected from naïve and inflamed rats. Markers assessing inflammation (i.e., weight changes and myeloperoxidase levels) were assessed on days 2, 7, 14 and 28 and compared to controls. Visceral hypersensitivity of the rats was assessed with colo-rectal distension and mechanical threshold testing on hind paws. IBS patients (n = 12) were age, gender, race, and BMI-matched with healthy controls (n = 12). Peripheral whole blood and plasma from fasting participants was collected and VIP plasma levels were assayed using a VIP peptide-enzyme immunoassay. Human gene expression of VIP was analyzed using a custom PCR array. TNBS induced colitis in the rats was confirmed with weight loss (13.7 ± 3.2 g) and increased myeloperoxidase activity. Visceral hypersensitivity to colo-rectal distension was increased in TNBS treated rats up to 21 d and resolved by day 28. Somatic hypersensitivity was also increased up to 14 d post TNBS induction of colitis. The expression of an inflammatory marker myeloperoxidase was significantly elevated in the intracellular granules of neutrophils in rat models following TNBS treatment compared to naïve rats. This confirmed the induction of inflammation in rats following TNBS treatment. VIP plasma concentration was significantly increased in rats following TNBS treatment as compared to naïve animals (P < 0.05). Likewise, the VIP gene expression from peripheral whole blood was significantly upregulated by 2.91-fold in IBS patients when compared to controls (P < 0.00001; 95%CI). VIP plasma protein was not significantly different when compared with controls (P = 0.193). Alterations in VIP expression may play a role in IBS. Therefore, a better understanding of the physiology of VIP could lead to new therapeutics.

Identification of local angiogenic and inflammatory markers in the menstrual blood of women with endometriosis

Evaluate the presence of inflammatory markers myeloperoxidase (MPO), N-acetylglucosaminidase (NAG), tumor necrosis factor alpha(TNF-α) and angiogenesis (vascular endothelial growth factor - VEGF) in peripheral and menstrual blood in women with and without endometriosis.

The common G-866A polymorphism of the UCP2 gene and survival in diabetic patients following myocardial infarction

BackgroundA variant in the promoter of the human uncoupling protein 2 (UCP2) gene, the G-866A polymorphism, has been associated with future risk of coronary heart disease events, in those devoid of traditional risk factors and in those suffering from diabetes. We thus examined the impact of the G-866A polymorphism on 5-year survival in a cohort of 901 post-myocardial infarction patients, and the impact of type-2 diabetes on this relationship. The association of UCP2 with baseline biochemical and hormonal measurements, including levels of the inflammatory marker myeloperoxidase, was also examined.MethodsUCP2 G-866A genotypes were determined using a polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) protocol. Myeloperoxidase levels were measured in plasma samples taken from 419 cohort patients 24–96 hours after admission.ResultsGenotypes were obtained for 901 patients with genotype frequencies AA 15.5%, GA 45.5%, and GG 39.0%. Genotype was not associated with survival in the overall cohort (mortality: AA 15.6%, GA 16.8%, GG 19.4%, p = 0.541). However, amongst diabetics, AA and GA genotype groups had significantly worse survival than GG diabetic patients (p < 0.05) with an attributable risk of 23.3% and 18.7% for those of AA and GA genotype respectively. Multivariate analysis using a Cox proportional hazards model confirmed that the interaction of diabetes with genotype was significantly predictive of survival (p = 0.031). In the cohort's diabetic subgroup AA/GA patients had higher myeloperoxidase levels than their GG counterparts (GA/AA, n = 51, 63.9 ± 5.23; GG, n = 34, 49.1 ± 3.72 ng/ml, p = 0.041). Further analysis showed that this phenomenon was confined to male patients (GA/AA, n = 36, 64.3 ± 6.23; GG, n = 29, 44.9 ± 3.72 ng/ml, p = 0.015).ConclusionDiabetic patients in this post-myocardial infarction cohort with UCP2 -866 AA/GA genotype have poorer survival and higher myeloperoxidase levels than their GG counterparts.

Abstract 3995: Comparison of the Effects of Prasugrel and High Dose Clopidogrel on In Vivo and In Vitro Platelet Activation: Results from PRINCIPLE-TIMI 44

In PRINCIPLE-TIMI 44, a randomized study in 201 patients undergoing cardiac catheterization for planned PCI, we reported that prasugrel resulted in greater inhibition of in vitro ADP-stimulated platelet aggregation than clopidogrel 600 mg loading dose (LD) and 150 mg maintenance dose (MD). Here we compare the effects of prasugrel and clopidogrel on in vivo and in vitro platelet activation. As prespecified end points of PRINCIPLE-TIMI 44, we measured circulating platelet-monocyte aggregates (PMA), circulating platelet-neutrophil aggregates (PNA), circulating P-selectin-positive platelets (PSPP), plasma soluble CD40 ligand (sCD40L) and (because activated platelets induce its release from leukocytes) myeloperoxidase (MPO). We also evaluated in vitro ADP-stimulated PMA, PNA and PSPP. At 6 h post LD, clopidogrel and prasugrel resulted in similar reductions in circulating PMA (Fig. A ), PNA and PSPP. In vitro ADP-stimulated PMA (Fig. B ), PNA and PSPP were inhibited to a greater degree by prasugrel than clopidogrel during the LD and MD phases. At 6 h post LD, clopidogrel- but not prasugrel-treated patients had elevated plasma MPO compared to pre-treatment (Fig. C ). Plasma sCD40L was unaffected by time or treatment. In this randomized study: Prasugrel, like clopidogrel, inhibited in vivo platelet activation. Prasugrel, to a greater degree than clopidogrel, inhibited ADP-stimulated PMA, PNA and PSPP – which may be mechanistically important, given the high local concentrations of ADP at sites of thrombus formation. Prasugrel, unlike clopidogrel, prevented the post-PCI-induced increase in the inflammatory marker MPO.

The effects of erdosteine, N‐acetylcysteine, and vitamin E on nicotine‐induced apoptosis of hippocampal neural cells

This study investigated the frequency of apoptosis in rat hippocampal neural cells after intraperitoneal nicotine injection, examining the roles of the inflammatory markers myeloperoxidase (MPO) and tumor necrosis factor alpha (TNF-alpha) in nicotine-induced brain damage and the protective effects of three known antioxidant agents, N-acetylcysteine (NAC), erdosteine, and vitamin E. Female Wistar rats were divided into seven groups, each composed of nine rats: 2 negative control groups, 2 positive control groups, one erdosteine-treated group (500 mg/kg), one NAC-treated group (500 mg/kg), and one vitamin E-treated group (500 mg/kg). Nicotine was intraperitoneally injected at a dosage of 0.6 mg/kg for 21 days. Following nicotine injection, the antioxidants were administered orally; treatment was continued until the rats were killed. Apoptosis level in hippocampal neural cells was determined by using TUNEL (terminal deoxynucleotidyl transferase-mediated dUTP nick endlabeling) method. Staining of cytoplasmic TNF-alpha in hippocampal neural cells and hippocampus MPO activity were evaluated by immunohistochemistry. Nicotine administration had no effect on local TNF-alpha production, or hippocampal MPO activity. The treatments with erdosteine, NAC and vitamin E significantly reduced the rate of nicotine-induced hippocampal neural cell apoptosis. This findings suggest that erdosteine and NAC can be as effective as vitamin E in protecting against nicotine-induced hippocampal neural cell apoptosis.

Macrophage‐specific LITAF (lipopolysaccharide induced TNF‐alpha factor) knockout mice (LITAF mac −/−) have a reduced inflammatory response to colonic administration of trinitrobenzene sulfonic acid (TNBS)

Dysregulation of TNF‐α secretion by lamina propria macrophages (LPM) is a hallmark of the inflammatory bowel disease Crohn's Disease (CD). LITAF has been shown to regulate TNF expression in macrophages and macrophage‐specific LITAF mac −/− mice (KO) are resistant to LPS induced sepsis. LITAF protein is increased by 30% in LPM isolated from C57BL/6 mice (WT) after TNBS administration, a well established model of CD (p&lt;0.05). The aim of this study was to compare the colonic inflammatory responses of WT mice to LITAF mac−/− mice after administration of TNBS.Methods: WT and KO mice were administered TNBS (3 mg in 50% EtOH) rectally. After 24 hrs, body weights (BW) were determined and levels of the inflammatory marker myeloperoxidase (MPO) and TNF‐α mRNA were measured in colon tissue.Results: TNBS administration reduces BW in all animals compared to untreated controls (p&lt;0.05); however, KO mice lost 50% less BW than the WT mice (p&lt;0.05). MPO activity is five fold lower in KO mice compared to WT (p&lt;0.05). Tissue levels of TNF‐α mRNA is also reduced in the KO compared to WT animals (376 ± 156 vs. 843± 253 copies) after TNBS administration.Conclusions:LITAF mac −/− have an attenuated inflammatory response to colonic TNBS administration. These results highlight a potential value of anti‐LITAF strategies for therapeutic management of CD. Research Support: BMC departments of Surgery and Pharmacology and NIDCR R01 DE014079.

Regulation of nicotine-induced apoptosis of pulmonary artery endothelial cells by treatment of N-acetylcysteine and vitamin E

This study investigated the frequency of apoptosis in rat pulmonary artery endothelial cells after intraperitoneal nicotine injection, examining the roles of the inflammatory markers myeloperoxidase (MPO), tumour necrosis factor alpha (TNF-alpha), and vascular endothelial growth factor (VEGF) in nicotine-induced vascular damage and the protective effects of two known antioxidant agents, N-acetylcysteine (NAC) and vitamin E. Female Wistar rats were divided into four groups, each composed of nine rats: negative control group, positive control group, NAC-treated group (500 mg/kg), and vitamin E-treated group (500 mg/kg). Nicotine was intraperitoneally injected at a dosage of 0.6 mg/kg for 21 days. Following nicotine injection, the antioxidants were administered orally; treatment was continued until the rats were killed. Lung tissue samples were stained with hematoxylin-eosin (H&E) for histopathological assessments. Apoptosis level in endothelial cells was determined by using TUNEL (terminal deoxynucleotidyl transferase-mediated dUTP nick end-labelling) method. Staining of cytoplasmic TNF-alpha and VEGF in endothelial cells, and perivascular MPO activity were evaluated by immunohistochemistry. The treatments with NAC and vitamin E significantly reduced the rate of nicotine-induced endothelial cell apoptosis. NAC and vitamin E significantly reduced the increases in the local production of TNF-alpha and VEGF, and perivascular MPO activity. This findings suggest that NAC can be as effective as vitamin E in protecting against nicotine-induced endothelial cell apoptosis.