Increase In Inflammatory Mediators Research Articles

Experimental spinal cord injury upregulates vagal nodose TRPV1 expression (902.4)

The gastrointestinal (GI) tract is critically dependent upon adequate blood flow. Spinal cord injury (SCI) produces dramatic GI and vascular dysregulation, including vascular hypotension and, possibly, visceral hypoperfusion. We have previously shown that T3‐SCI in rats results in diminished visceral blood flow, increased expression of inflammatory markers and consequent gastrointestinal dysmotility. Vagal afferent injury has been shown to induce changes in cholecystokinin (CCK) and transient receptor potential vanilloid type 1 receptor (TRPV1) expression within the nodose ganglion. We hypothesized that SCI leads to increased activation of TRPV1 channels along the vagus nerve.Nodose ganglia TRPV1 and CCK mRNA was quantified using qRT‐PCR, at 1‐day, 3‐days, and 7‐days post‐SCI and control. We used a superior mesenteric artery (SMA) occlusion model to show a worst case scenario of ischemia‐reperfusion to the GI tract. Our data show a significant increase in nodose TRPV1 and CCK of SCI and SMA rats compared to controls. The increased TRPV1 indicates GI inflammation and the increased CCK suggests that CCK increases as per vagal afferent injury. Our data present similarities to experimental ischemia‐reperfusion of the GI tract, in which SMA occlusion causes a dramatic inflammatory response within the GI tract. We propose that our model of SCI represents a continuum of GI ischemia and that the increase in inflammatory mediators and TRPV1 activation may play a role in the post‐SCI pathophysiology of gastric vagal afferents.Grant Funding Source: Supported by: NINDS 49177

Anti‐inflammatory effects of cannabinoid CB2 receptor activation in endotoxin‐induced uveitis

Cannabinoid CB2 receptors mediate immunomodulation. Here, we investigated the effects of CB2 receptor ligands on leukocyte-endothelial adhesion and inflammatory mediator release in experimental endotoxin-induced uveitis (EIU). EIU was induced by intraocular injection of lipopolysaccharide (LPS, 20 ng·μL(-1) ). Effects of the CB2 receptor agonist, HU308 (1.5% topical), the CB2 receptor antagonist, AM630 (2.5 mg·kg(-1) i.v.), or a combination of both compounds on leukocyte-endothelial interactions were measured hourly for 6 h in rat iridial vasculature using intravital microscopy. Anti-inflammatory actions of HU308 were compared with those of clinical treatments for uveitis - dexamethasone, prednisolone and nepafenac. Transcription factors (NF-κB, AP-1) and inflammatory mediators (cytokines, chemokines and adhesion molecules) were measured in iris and ciliary body tissue. Leukocyte-endothelium adherence was increased in iridial microvasculature between 4-6 h after LPS. HU308 reduced this effect after LPS injection and decreased pro-inflammatory mediators: TNF-α, IL-1β, IL-6, CCL5 and CXCL2. AM630 blocked the actions of HU-308, and increased leukocyte-endothelium adhesion. HU-308 decreased levels of the transcription factors NF-κB and AP-1, while AM630 increased levels of NF-κB. Topical treatments with dexamethasone, prednisolone or nepafenac, failed to alter leukocyte adhesion or mitigate LPS-induced increases in inflammatory mediators during the 6 h of EIU. Activation of CB2 receptors was anti-inflammatory in a model of acute EIU and involved a reduction in NF-κB, AP-1 and inflammatory mediators. CB2 receptors may be promising drug targets for the development of novel ocular anti-inflammatory agents. This article is part of a themed section on Cannabinoids 2013. To view the other articles in this section visit http://dx.doi.org/10.1111/bph.2014.171.issue-6.

Erythropoietin pretreatment suppresses seizures and prevents the increase in inflammatory mediators during pentylenetetrazole-induced generalized seizures

Erythropoietin (EPO) suppresses epileptic seizures, but the mechanism is unclear. The search for novel targets in the therapy of epilepsy has focused recently on brain inflammation since brain inflammation and the associated blood–brain barrier (BBB) damage appears to be an integral part of epilepsy pathophysiology. We examined the effects of EPO on proinflammatory mediators in brain and serum in PTZ-induced generalized seizure model. The inflammation markers (IL-1β, TNF-α, IL-6, IL-10), BBB and neuron damage markers (S100B, Neuron specific enolase; NSE, respectively) in serum and brain of Sprague–Dawley male rats were examined with the ELISA method. Nitric oxide synthase (NOS) isoforms were investigated immunohistochemically in hippocampus. EPO treatment 4 h and 24 h before PTZ administration had diverse effects. EPO treatment 4 h before PTZ administration elongated the seizure latency, decreased the inflammation and damage markers in serum and brain significantly, whereas EPO treatment 24 h before PTZ administration lowered inflammation and damage markers to control levels and decreased the seizure stage. PTZ-induced seizures increased inducible NOS (iNOS) activity and decreased endothelial NOS (eNOS) activity in hippocampus. Both EPO pretreatments reversed these effects. These findings, i.e., decreased iNOS activity and increased eNOS activity by EPO suggest the first time that the favorable effect of EPO pretreatment on inflammatory mediators triggered by PTZ-induced seizures. This can provide further insight into epilepsy treatment and new prophylactic strategies against epilepsy risk.

Mitochondrial Dysfunction and Tissue Alterations of Ultraviolet-Irradiated Skin in Five Different Mice Strains

Ultraviolet radiation (UVR) effects on skin have been extensively studied. Several mice strains have been used worldwide in photobiology and photoimmunology studies. Recently, we have developed a method based on flow cytometry in order to analyze mitochondrial dysfunction and superoxide (O 2 • - ) production ex vivo in keratinocytes isolated from irradiated-mice. This method can be helpful to evaluate mitochondrial alterations in different mice models. In this work, we aimed to compare epidermal response to UVR in five mice strains, both pigmented and albino as well as hairy and hairless strains (SKH:1, Balb/c, C57/BL, DBA/2N and Swiss). Keratinocytes mitochondrial alterations, epidermal hyperplasia and inflammatory mediators’ production (epidermis and serum) were determined 72 hours after a 400 mJ/cm 2 UV dose. All strains showed epidermal hyperplasia and loss of mitochondrial polarization after irradiation, differing in the magnitude of the response. However, there were significant differences in the basal mitochondrial polarization between strains, showing that the metabolic state of keratinocytes may vary between them. Moreover, mitochondrial O 2 • - production was induced in SKH:1 and Balb/c after irradiation, whereas in DBA/2N, Swiss and C57/BL it was at the same level or even lower than in the non-irradiated control. Finally, an increase in inflammatory mediators was only detected in the serum of C57/BL and Swiss mice and in the epidermis of DBA/2N and C57/BL. Results show that each mice strain has particular characteristics related to cellular metabolism, which may lead to particular responses to UVR exposure. Therefore, the use of a particular mice strain in photobiology models should be carefully considered.

RETRACTED: Increases in inflammatory mediators in DRG implicate in the pathogenesis of painful neuropathy in Type 2 diabetes

This article has been retracted: please see Elsevier Policy on Article Withdrawal (https://www.elsevier.com/about/our-business/policies/article-withdrawal). This article has been retracted at the request of the Editor-in-Chief. Image duplication has been observed within Figure 1g-h. The corresponding author has been asked to provide an acceptable explanation for this duplication but has not been able to do so neither have the original source files been supplied.

Levels of nitric oxide synthase and cholecystokinin mRNA in the upper gastrointestinal tract of rats following experimental spinal cord injury

In addition to the loss of motor and sensory function, SCI causes immediate changes to gastrointestinal (GI) tract physiology and reflex function. Reflex control of the stomach is dominated by vago‐vagal circuits which remain anatomically intact following SCI. Our previous reports indicate that SCI a) reduces gastric motility, tone, and emptying; b) diminishes vagal afferent sensitivity to GI peptides such as cholecystokinin (CCK); and c) diminishes mesenteric blood flow.The mechanisms, interrelationships, and temporal patterning of post‐SCI gastroparesis remain poorly defined. We tested the hypothesis that acute SCI induces GI inflammation and a resulting reduction of CCK mRNA expression.We used qRT‐PCR to quantify the levels of CCK, iNOS, and nNOS in the GI tract, 1 week post‐SCI. Our data show a significant increase in CCK and iNOS in the duodenum; nNOS shows an upward non‐significant trend. Jejunal expression of CCK is reduced while both iNOS and nNOS levels show a non‐significant upregulation. Our data are similar to experimental ischemia‐reperfusion of the GI tract, in which the superior mesenteric artery (SMA) occlusion causes a dramatic inflammatory response within the GI tract. We conclude that our model of SCI represents a continuum of GI ischemia and that the increase in inflammatory mediators may play a role in decreasing the sensitivity of the gastric vagal afferents.Grant Funding Source: NINDS 49177

Aging accentuates alcohol-induced decrease in protein synthesis in gastrocnemius

The present study sought to determine whether the protein catabolic response in skeletal muscle produced by chronic alcohol feeding was exaggerated in aged rats. Adult (3 mo) and aged (18 mo) female F344 rats were fed a nutritionally complete liquid diet containing alcohol (36% of total calories) or an isocaloric isonitrogenous control diet for 20 wk. Muscle (gastrocnemius) protein synthesis, as well as mTOR and proteasome activity did not differ between control-fed adult and aged rats, despite the increased TNF-α and IL-6 mRNA and decreased IGF-I mRNA in muscle of aged rats. Compared with alcohol-fed adult rats, aged rats demonstrated an exaggerated alcohol-induced reduction in lean body mass and protein synthesis (both sarcoplasmic and myofibrillar) in gastrocnemius. Alcohol-fed aged rats had enhanced dephosphorylation of 4E-BP1, as well as enhanced binding of raptor with both mTOR and Deptor, and a decreased binding of raptor with 4E-BP1. Alcohol feeding of both adult and aged rats reduced RagA binding to raptor. The LKB1-AMPK-REDD1 pathway was upregulated in gastrocnemius from alcohol-fed aged rats. These exaggerated alcohol-induced effects in aged rats were associated with a greater decrease in muscle but not circulating IGF-I, but no further increase in inflammatory mediators. In contrast, alcohol did not exaggerate the age-induced increase in atrogin-1 and MuRF1 mRNA or the increased proteasome activity. Our results demonstrate that, compared with adult rats, the gastrocnemius from aged rats is more sensitive to the catabolic effects of alcohol on protein synthesis, but not protein degradation, and this exaggerated response may be AMPK-dependent.

Kolaviron, a Natural Antioxidant and Anti‐Inflammatory Phytochemical Prevents Dextran Sulphate Sodium‐Induced Colitis in Rats

The beneficial effects of kolaviron, a natural biflavonoid from the seeds of Garcinia kola, have been attributed mainly to its antioxidant and anti-inflammatory effects. This study investigated these effects on dextran sulphate sodium (DSS)-induced ulcerative colitis in rats. Sulfasalazine served as standard reference in this study. Kolaviron and sulfasalazine were separately co-administered orally at 200 mg/kg and 500 mg/kg, respectively, to dextran sulphate sodium-exposed rats for 5 days. The result indicated that kolaviron or sulfasalazine significantly prevented DSS-induced body weight loss as well as the incidence of diarrhoea and bleeding in DSS-exposed rats. Kolaviron suppressed the DSS-mediated increase in colonic nitric oxide concentration and myeloperoxidase activity and significantly prevented the increase in inflammatory mediators, interleukin-1β and tumour necrosis factor alpha, in the colon of DSS-treated rats. The significant depletion in colonic antioxidant status in rats exposed to DSS alone was evident by marked reduction in colonic catalase and glutathione S-transferase activities as well as glutathione content, leading to elevated hydrogen peroxide and lipid peroxidation levels. Histopathologically, DSS alone resulted in severe epithelial erosion, total absence of goblet cells, destruction of the crypts, necrotic and distorted glands, accompanied by marked cellular mononuclear cells infiltration. However, administration of kolaviron and sulfasalazine ameliorated DSS-induced colitis by increasing the antioxidant status decreased hydrogen peroxide and lipid peroxidation levels and attenuated the adverse effect of DSS on colon architecture. In conclusion, the anti-colitis effect of kolaviron is related to its intrinsic anti-inflammatory and anti-oxidative properties.

New Technique in Tendon Sport Recovery. Percutaneous Electrolysis Intratissue (EPI®)

Based on the literature and clinical experience we know that the technique of percutaneous electrolysis intratissue (EPI ® ) has positive effects on the recovery of human tendinopathies. The application of the technique, together with the completion of eccentric exercise, give benefits leading to anatomical and functional recovery in the that can be appreciated by ultrasound using the scale of Victorian Institute of Sport Assessment-patellar tendon (VISA-P). Due to clinical knowledge, damage to is accompanied by increase in oxidative stress, cell death and inflammation at the damaged area. The EPI ® technique produced a galvanic current inside the tendon, with an acupuncture applicator, producing increase in cell death, inflammation and oxidative stress in the first days of treatment. After these days, the recovery of is quickly and higher than without EPI ® technique used. Galvanic current is produced in a salad solution producing chemical reaction with salt decomposition (NaCl) and H 2 O 2 in its chemical constituent elements. After they form new substances, such as NaOH, H 2 and Cl 2 , they change the reactions inside the damage. The NaOH has vital importance because it is high caustic and destroy collagen and mixed substances in damage area. EPI ® is a basic technique giving a chemist process without boil electrocution of tissue. When EPI ® is used, an increase in inflammatory response of the tissue is also detected. In chronic tendon, with the unique possibility of recovery by surgery, EPI ® technique has recuperation of without surgery. EPI ® technique, in European countries, has been used in the last decade with wonderfully results to recovery pathological tendons from athletes in competition. Furthermore, it is known that after recovery from injury in damaged area, neo-vascularization occurs. It is intent of to obtain more energy to recover damage tissue. Valles and its collaborators have demonstrated the molecular mechanism of action of EPI ® technique (1). First of all, an increase in oxidative stress and inflammation occurs in the chronic damage. After, we appreciate a neovascularization in the damage area with increase in inflammatory mediators. As we know in damage tendon, in first days of lesion, an increase in inflammation and vascularization are detected, but sometimes recovery occurs with a non-excellent distribution of the tissue. With the EPI ® ® technique. In research with rats, we have demonstrated an increase in vascular endothelial growth factor (VEGF) and its receptor VEGFR-1 and 2, demonstrating an increase in neovascularization in the damage area. Also we detect, days after application of EPI ® technique an increase in PPAR-γ production, anti- inflammatory protein produced after inflammation occurs in damage tissues. With all, we want to note here the importance of this technique in the future to recover damage in sport population and in football players in particular.

In VivoTLR9 Inhibition Attenuates CpG-Induced Myocardial Dysfunction

The involvement of toll-like receptor 9 (TLR9), a receptor for bacterial DNA, in septic cardiac depression has not been clarified in vivo. Thus, the aim of the study was to test possible TLR9 inhibitors (H154-thioate, IRS954-thioate, and chloroquine) for their ability to protect the cardiovascular system in a murine model of CpG oligodeoxynucleotide- (ODN-) dependent systemic inflammation. Sepsis was induced by i.p. application of the TLR9 agonist 1668-thioate in C57BL/6 wild type (WT) and TLR9-deficient (TLR9-D) mice. Thirty minutes after stimulation TLR9 antagonists were applied i.v. Survival was monitored up to 18 h after stimulation. Cardiac mRNA expression of inflammatory mediators was analyzed 2 h and 6 h after stimulation with 1668-thioate and hemodynamic parameters were monitored at the later time point. Stimulation with 1668-thioate induced a severe sepsis-like state with significant drop of body temperature and significantly increased mortality in WT animals. Additionally, there was a time-dependent increase of inflammatory mediators in the heart accompanied by development of septic heart failure. These effects were not observed in TLR9-D mice. Inhibition of TLR9 by the suppressive ODN H154-thioate significantly ameliorated cardiac inflammation, preserved cardiac function, and improved survival. This suppressive ODN was the most efficient inhibitor of the tested substances.

ISSLS Prize Winner

Animal model of intravertebral disc (IVD) degeneration. To examine production of inflammatory mediators in IVDs and neuropeptides in dorsal root ganglia (DRGs) in rat models of IVD compression and injury. Sensory nerve fibers in IVDs and inflammatory mediator responses have been verified in animal models of IVD injury. However, the IVD injury in animals incompletely models degenerated human IVDs causing discogenic low back pain, because human IVDs are also subject to compression. Experimental groups (controls, IVD injury, IVD compression, and their combination) of Sprague Dawley rats were prepared. Fluoro-Gold (FG; Fluorochrome, Denver, CO) was applied into coccygeal IVDs. Inflammatory mediators in IVDs, including nerve growth factor, tumor necrosis factor α, interleukin 1β, and interleukin 6, were quantified using enzyme-linked immunosorbent assays. DRGs were immunostained for calcitonin gene-related peptide, activating transcription factor 3, and growth-associated phosphoprotein 43. The upregulation of inflammatory mediators was transient in the IVD injury group but delayed and long-lasting in the IVD compression group. When the IVD injury and compression were combined, the upregulation of inflammatory mediators was long-lasting through 8 weeks. The proportion of calcitonin gene-related peptide-immunoreactive neurons among Fluoro-Gold-labeled neurons remained significantly higher in the IVD injury, compression, and combination groups than in the controls. In contrast, increases in the proportions of activating transcription factor 3-immunoreactive or growth-associated phosphoprotein 43-immunoreactive neurons in the IVD injury group animals were transient but long-lasting in the compression and combination groups compared with controls. Disc injury in rats produces persistent increases in neuropeptides in DRGs but only transient increases in inflammatory mediators in IVDs. On the contrary, disc compression in rats produces a long-lasting increase in inflammatory mediators in IVDs and neuropeptides in DRGs. Moreover, disc compression induces persistent nerve injury and regeneration of the afferent fibers innervating IVDs.

Comparative Study of Systemic Inflammatory Responses in Psoriasis Vulgaris and Mild to Moderate Allergic Contact Dermatitis

Background: Psoriasis vulgaris (PV) is associated with low-grade systemic inflammation. Objective: Cytokines’ and growth factors’ serum patterns in patients with PV, allergic contact dermatitis (ACD) and healthy subjects were investigated to describe and compare systemic inflammatory responses in these diseases. Methods: A total of 12 inflammation-sensitive biomarkers were analyzed simultaneously by means of the Evidence Investigator™ biochip technology. Results: In PV, proinflammatory tumor necrosis factor (TNF)-α, interferon (IFN)-γ, interleukins (IL)-1β, -2, -6, -8, and monocyte chemoattractant protein (MCP)-1 were elevated. In ACD, 2 markers, TNF-α and MCP-1, were increased, and regulatory cytokine IL-10 was decreased. Proinflammatory IL-2 had the strongest correlations with other pro- as well as anti-inflammatory cytokines in PV and ACD, whilst IL-6 correlated positively with the Psoriasis Area and Severity Index. Growth factors’ levels correlated with MCP-1, but only in PV. Conclusion: Although psoriasis induces a more variegated proinflammatory systemic response, ACD is likewise associated with a systemic increase in inflammatory mediators.

Lumbar facet joint compressive injury induces lasting changes in local structure, nociceptive scores, and inflammatory mediators in a novel rat model.

Objective. To develop a novel animal model of persisting lumbar facet joint pain. Methods. Sprague Dawley rats were anaesthetized and the right lumbar (L5/L6) facet joint was exposed and compressed to ~1 mm with modified clamps applied for three minutes; sham-operated and naïve animals were used as control groups. After five days, animals were tested for hind-paw sensitivity using von Frey filaments and axial deep tissue sensitivity by algometer on assigned days up to 28 days. Animals were sacrificed at selected times for histological and biochemical analysis. Results. Histological sections revealed site-specific loss of cartilage in model animals only. Tactile hypersensitivity was observed for the ipsi- and contralateral paws lasting 28 days. The threshold at which deep tissue pressure just elicited vocalization was obtained at three lumbar levels; sensitivity at L1 > L3/4 > L6. Biochemical analyses revealed increases in proinflammatory cytokines, especially TNF-α, IL-1α, and IL-1β. Conclusions. These data suggest that compression of a facet joint induces a novel model of local cartilage loss accompanied by increased sensitivity to mechanical stimuli and by increases in inflammatory mediators. This new model may be useful for studies on mechanisms and treatment of lumbar facet joint pain and osteoarthritis.

Pathogenic Mechanisms Linking Periodontal Diseases With Adverse Pregnancy Outcomes

In the last 2 decades, a large proportion of studies have focused on the relationship between maternal periodontal disease and poor obstetric outcomes. The aim of the present review is to summarize the current knowledge about human studies on the pathogenetic mechanisms linking periodontal diseases with adverse pregnancy outcomes. A search of the medical literature was conducted using NIH (National Institute of Health) Pubmed through April 2011. Articles were identified with the Medical Subject Heading (MeSH) and free text terms "small for gestational age (SGA)," "preeclampsia," "preterm labor," and "periodontal disease." Experimental human studies have shown that periodontal pathogens may disseminate toward placental and fetal tissues accompanied by an increase in inflammatory mediators in the placenta. As such, new inflammatory reactions within the placental tissues of the pregnant woman may occur, the physiological levels of prostaglandin E(2) (PGE(2)) and tumor necrosis factor-α (TNF-α) in the amniotic fluid may increase and eventually lead to premature delivery. Although many data from clinical trials suggest that periodontal disease may increase the adverse pregnancy outcome, the exact pathogenetic mechanism involved remains controversial. The findings explain the potential link between periodontal infections and adverse pregnancy outcomes. First, periodontal bacteria can directly cause infections both of the uteroplacenta and the fetus; second, systemic inflammatory changes induced by periodontal diseases can activate responses at the maternal-fetal interface. Of note, associative studies have produced different results in different population groups and no conclusive evidence has still been produced for the potential role of preventive periodontal care to reduce the risk factors of preterm birth.

Systemic Inflammatory Response Does Not Correlate with Acute Lung Injury Associated with Mechanical Ventilation Strategies in Normal Lungs

Mechanical ventilation (MV) can lead to ventilator-induced lung injury secondary to trauma and associated increases in pulmonary inflammatory cytokines. There is controversy regarding the associated systemic inflammatory response. In this report, we demonstrate the effects of MV on systemic inflammation. This report is part of a previously published study (Hong et al. Anesth Analg 2010;110:1652-60). Female pigs were randomized into 3 groups. Group H-Vt/3 was ventilated with a tidal volume (Vt) of 15 mL/kg predicted body weight (PBW)/positive end-expiratory pressure (PEEP) of 3 cm H(2)O; group L-Vt/3 with a Vt of 6 mL/kg PBW/PEEP of 3 cm H2O; and group L-Vt/10 with a Vt of 6 mL/kg PBW/PEEP of 10 cm H(2)O, for 8 hours. Each group had 6 subjects (n = 6). Prelung and postlung sera were analyzed for inflammatory markers. Hemodynamics, airway mechanics, and arterial blood gases were monitored. There were no significant differences in systemic cytokines among groups. There were similar trends of serum inflammatory markers in all subjects. This is in contrast to findings previously published demonstrating increases in inflammatory mediators in bronchoalveolar lavage. Systemic inflammatory markers did not correlate with lung injury associated with MV.

Neuroprotection by inhibiting the c-Jun N-terminal kinase pathway after cerebral ischemia occurs independently of interleukin-6 and keratinocyte-derived chemokine (KC/CXCL1) secretion

BackgroundCerebral ischemia is associated with the activation of glial cells, infiltration of leukocytes and an increase in inflammatory mediators in the ischemic brain and systemic circulation. How this inflammatory response influences lesion size and neurological outcome remains unclear. D-JNKI1, an inhibitor of the c-Jun N-terminal kinase pathway, is strongly neuroprotective in animal models of stroke. Intriguingly, the protection mediated by D-JNKI1 is high even with intravenous administration at very low doses with undetectable drug levels in the brain, pointing to a systemic mode of action, perhaps on inflammation.FindingsWe evaluated whether D-JNKI1, administered intravenously 3 h after the onset of middle cerebral artery occlusion (MCAO), modulates secretion of the inflammatory mediators interleukin-6 and keratinocyte-derived chemokine in the plasma and from the spleen and brain at several time points after MCAO. We found an early release of both mediators in the systemic circulation followed by an increase in the brain and went on to show a later systemic increase in vehicle-treated mice. Release of interleukin-6 and keratinocyte-derived chemokine from the spleen of mice with MCAO was not significantly different from sham mice. Interestingly, the secretion of these inflammatory mediators was not altered in the systemic circulation or brain after successful neuroprotection with D-JNKI1.ConclusionsWe demonstrate that neuroprotection with D-JNKI1 after experimental cerebral ischemia is independent of systemic and brain release of interleukin-6 and keratinocyte-derived chemokine. Furthermore, our findings suggest that the early systemic release of interleukin-6 and keratinocyte-derived chemokine may not necessarily predict an unfavorable outcome in this model.

Inhibition of Hydrogen Sulfide-induced Angiogenesis and Inflammation in Vascular Endothelial Cells: Potential Mechanisms of Gastric Cancer Prevention by Korean Red Ginseng

Previously, we reported that Helicobacter pylori-associated gastritis and gastric cancer are closely associated with increased levels of hydrogen sulfide (H2S) and that Korean red ginseng significantly reduced the severity of H. pylori-associated gastric diseases by attenuating H2S generation. Because the incubation of endothelial cells with H2S has been known to enhance their angiogenic activities, we hypothesized that the amelioration of H2S-induced gastric inflammation or angiogenesis in human umbilical vascular endothelial cells (HUVECs) might explain the preventive effect of Korean red ginseng on H. pylori-associated carcinogenesis. The expression of inflammatory mediators, angiogenic growth factors, and angiogenic activities in the absence or presence of Korean red ginseng extracts (KRGE) were evaluated in HUVECs stimulated with the H2S generator sodium hydrogen sulfide (NaHS). KRGE efficiently decreased the expression of cystathionine β-synthase and cystathionine γ-lyase, enzymes that are essential for H2S synthesis. Concomitantly, a significant decrease in the expression of inflammatory mediators, including cyclooxygenase-2 and inducible nitric oxide synthase, and several angiogenic factors, including interleukin (IL)-8, hypoxia inducible factor-1a, vascular endothelial growth factor, IL-6, and matrix metalloproteinases, was observed; all of these factors are normally induced after NaHS. An in vitro angiogenesis assay demonstrated that NaHS significantly increased tube formation in endothelial cells, whereas KRGE pretreatment significantly attenuated tube formation. NaHS activated p38 and Akt, increasing the expression of angiogenic factors and the proliferation of HUVECs, whereas KRGE effectively abrogated this H2S-activated angiogenesis and the increase in inflammatory mediators in vascular endothelial cells. In conclusion, KRGE was able to mitigate H2S-induced angiogenesis, implying that antagonistic action against H2S-induced angiogenesis may be the mechanism underlying the gastric cancer preventive effects of KRGE in H. pylori infection.

Disturbed One-Carbon Metabolism Causing Adverse Reproductive Outcomes in Mice Is Associated with Altered Expression of Apolipoprotein AI and Inflammatory Mediators PPARα, Interferon-γ, and Interleukin-10 ,

Low dietary choline or deficiency of methylenetetrahydrofolate reductase (Mthfr) leads to hyperhomocysteinemia (Hhcy) and adverse reproductive outcomes. Homocysteine reduces synthesis of ApoAI, the major lipoprotein in HDL-cholesterol; ApoAI is regulated by PPARα and has antiinflammatory properties. Our aim was to determine whether pregnancy complications due to genetic or nutritional deficiencies in 1-carbon metabolism could relate to dysregulation of ApoAI and inflammatory mediators. We fed pregnant mice, with or without a deficiency of Mthfr, control or choline-deficient (ChDD) diets for 10–12 wk and examined levels of ApoAI, PPARα, IFNγ, and IL-10. ApoAI mRNA was reduced in livers of Mthfr+/- mice and ApoAI protein was reduced due to Mthfr deficiency or choline deficiency in liver and plasma. Placental ApoAI protein was also reduced due to Mthfr genotype or choline-deficient diet and in developmentally delayed embryos. Reduced liver PPARα expression (mRNA and protein) was observed in ChDD-fed mice and was associated with increased methylation of a CpG dinucleotide in its promoter. Hepatic IFNγ increased due to genotype, and placental IFNγ was higher in Mthfr+/- ChDD-fed dams compared to Mthfr+/+ mice fed ChDD or Mthfr+/- mice fed CD. IL-10 was reduced in livers of ChDD-fed mice. We propose that a deficiency of dietary choline or Mthfr leads to Hhcy and reduced expression of maternal ApoAI, with reduced ApoAI transfer to embryo. Disturbances in 1-carbon metabolism also reduce maternal PPARα expression, possibly through promoter hypermethylation, and increase IFNγ and decrease IL-10 levels. This disturbance of the T helper (Th1) (IFNγ):Th2 (IL-10) ratio and the increase in inflammatory mediators may contribute to pregnancy complications.

Disk Injury in Rats Produces Persistent Increases in Pain-Related Neuropeptides in Dorsal Root Ganglia and Spinal Cord Glia but Only Transient Increases in Inflammatory Mediators

Immunohistological analysis in an injured intervertebral disk (IVD) model. To elucidate and compare in rats the behavior of the sensory nervous system and inflammatory mediators in experimentally injured IVDs. Multiple human and animal studies have verified the presence of sensory nerve fibers in IVDs or investigated the behavior of inflammatory mediators in injured IVDs, but no in vivo study to date has examined the relationship between the 2. Eight-week-old female rats were used. In the disk-injured group, L5/L6 disks were injured with a 24-gauge needle; simultaneously, the neurotracer Fluoro-gold was injected into the L5/L6 IVD. The L5/L6 IVD dorsal root ganglia (DRGs) from the L1 to L6 levels, and the spinal cord was resected at several time points after surgery. Nerve growth factor, tumor necrosis factor (TNF)-α and interleukin (IL)-6 production in the IVDs were quantified using enzyme-linked immunosorbent assay. DRGs were immunostained for calcitonin gene-related peptide, and spinal cord sections were immunostained for ionized calcium-binding adaptor molecule-1 and glial fibrillary acidic protein. Nerve growth factor, and TNF-α levels (through 1 week) and IL-6 levels (through 4 days) were significantly higher in the disk-injured group than in the noninjured group (P < 0.05). However, starting at 2 weeks (nerve growth factor and TNF-α) or 1 week (IL-6), the differences in inflammatory mediator levels between the 2 groups no longer were significant. In contrast, the percentage of calcitonin gene-related peptide-immunoreactive neurons among Fluoro-gold-labeled DRG neurons, and the numbers of ionized calcium-binding adaptor molecule-1-immunoreactive microglia and glial fibrillary acidic protein-immunoreactive astrocytes in the spinal dorsal horn remained significantly higher in the injured group than in the noninjured group at all-time points (P < 0.05). Disk injury in rats produces persistent increases in neuropeptides in DRGs and glia in the spinal cord, but only transient increases in inflammatory mediators in IVDs.

Does the observation of solution induced corneal staining signal the presence of corneal inflammation?

Purpose: To examine the differences in lipids deposits from worn silicone hydrogel materials (Balafilcon A and Lotrafilcon A) worn daily vs overnight. The deposits were analysed using Gas Chromatography Mass Spectrometry (GCMS). Method: Single lenses were extracted in chloroform: methanol (1:1) and transmethylated prior to analysis by GCMS. The lenses used were collected from a clinically controlled study (n=30) where lenses were worn daily or continuously for 30 days. Results: Lotrafilcon A lenses (16:0 average peak height 112 Kcounts) accumulated much less lipid than Balafilcon A (16:0 average peak height 1213 Kcounts). Results also showed a significant difference in lipid profile from lenses worn on a daily wear basis compared to those worn on a continuous wear basis and this was shown to be irrespective of lens material. The lenses that were worn daily for a month accumulated more unsaturated lipid such as C16:1 and C18:1. The lenses which had been worn continuously for the same period of time had little to no unsaturated fatty acids. Conclusions: Although atmospheric oxidation of lipids occurs during daily wear of contact lenses, oxidation of lens-immobilised lipids during successive periods of overnight wear is even more dramatic. The fact that lenses worn on an overnight basis for 30 days yielded predominately saturated lipids suggests that unsaturated lipids are oxidised during eye closure. This correlates with the known increase in inflammatory mediators in the overnight tear film and indicates a marked increase in free radical production leading to the degradation of unsaturated lipids.