Polymorphonuclear Leukocyte Infiltration Research Articles

The role of trimetazidine in ischemia/reperfusion damage treatment in an ovary torsion model experimentally induced in rats

The aim of this experimental animal study was to investigate the histopathological and biochemical efficacy of trimetazidine (TMZ) in decreasing ovary damage in an ovary ischaemia/reperfusion (I/R) model in the rat. A total of 35 Wistar albino female rats were randomly separated into five groups, n = 7 per group: Group 1: Sham (S) was only given a laparotomy procedure. Group 2: Ischaemia (I) group with 2-hour ischaemia using a vascular sutur. Group 3: Ischaemia/Reperfusion (I/R) group with 2 hour ischaemia and 2-hour reperfusion. Group 4: Sham + 10 mg/kg orally TMZ (S + TMZ). Group 5: I/R + 10 mg/kg oral TMZ (I/R + TMZ) group with 2 hours ischaemia and 2 hours reperfusion after the administration orally 10 mg/kg oral TMZ. Two daily doses of TMZ were orally administered to Group 4 (S + TMZ) and Group 5 (I/R + TMZ) for three days. TMZ significantly decreased vascular congestion, haemorrhage, and polymorphonuclear leukocyte infiltration in group 5 compared to group 3 (p < .05). Despite TMZ decreased the malondialdehyde, total oxidant status, and oxidative stress index values, these decreases were not statistically significant (p > .05). TMZ which is an antioxidant agent can efficiently prevent in I/R damage in rat ovaries but further studies are necessary in order to implement it in the clinical settings. IMPACT STATEMENT What is already known on this subject? Adnexial torsion is the most common gynecological emergency and there are no specific clinical, laboratories, or radiological findings for adnexal torsion. Unfortunatelly, the currently accepted treatment is adnexal detorsion. Cytoprotective effects of Trimetazidine (TMZ), an antianginal drug, are well-defined and it has been demonstrated to improve oxidative stress markers and limits membrane damage induced by reactive oxygen species and protects tissues from free radicals with its antioxidant effects. The aim of this study is to investigate the effects of TMZ in experimentally induced adnexal torsion in rats and to investigate possible effects in maintaining ovarian reserve to prevent I/R damage or reperfusion damage. What do the results of this study add? Our study showed that TMZ significantly decreased vascular congestion, haemorrhage, and PMNL infiltration. TMZ decreased the malondialdehyde, total oxidant status, and the oxidative stress index values, but these decreases were not statistically significant. What are the implications of these findings for clinical practice and/or further research? Although various antioxidant drugs and chemicals have been used to protect the ovaries against I/R damage, they have not been demostrated to prevent it completely. TMZ, an antioxidant efficacy agent, has been shown to prevent ovarian I/R damage by suppressing inflammation in terms of histopathological parameters. Further studies involving a greater number of experimental animals are required before using TMZ for the treatment of humans with I/R damage in the clinical setting.

IL-17 Aggravates Pseudomonas aeruginosa Airway Infection in Acute Exacerbations of Chronic Obstructive Pulmonary Disease.

Pseudomonas aeruginosa airway infection increases risks of exacerbations and mortality in chronic obstructive pulmonary disease (COPD). We aimed to elucidate the role of IL-17 in the pathogenesis. We examined the expression and influences of IL-23/IL-17A in patients with stable COPD (n = 33) or acute COPD exacerbations with P. aeruginosa infection (n = 34). A mouse model of COPD (C57BL/6) was used to investigate the role of IL-17A in host inflammatory responses against P. aeruginosa infection through the application of IL-17A–neutralizing antibody or recombinant IL-17A. We found that P. aeruginosa infection increased IL-23/17A signaling in lungs of both COPD patients and COPD mouse models. When COPD mouse models were treated with neutralizing antibody targeting IL-17A, P. aeruginosa induced a significantly less polymorphonuclear leukocyte infiltration and less bacterial burden in their lungs compared to those of untreated counterparts. The lung function was also improved by neutralizing antibody. Furthermore, IL-17A-signaling blockade significantly reduced the expression of pro-inflammatory cytokine IL-1β, IL-18, TNF-α, CXCL1, CXCL15 and MMP-9, and increased the expression of anti-inflammatory cytokine IL-10 and IL-1Ra. The application of mouse recombinant IL-17A exacerbated P. aeruginosa-mediated inflammatory responses and pulmonary dysfunction in COPD mouse models. A cytokine protein array revealed that the expression of retinol binding protein 4 (RBP4) was down-regulated by IL-17A, and exogenous RBP4-recombinant protein resulted in a decrease in the severity of P. aeruginosa-induced airway dysfunction. Concurrent application of IL-17A-neutralizing antibody and ciprofloxacin attenuated airway inflammation and ventilation after inoculation of P. aeruginosa in COPD mouse models. Our results revealed that IL-17 plays a detrimental role in the pathogenesis of P. aeruginosa airway infection during acute exacerbations of COPD. Targeting IL-17A is a potential therapeutic strategy in controlling the outcomes of P. aeruginosa infection in COPD patients.

Neuropharmacological Alterations by a Rice Contaminant Stenotrophomonas maltophilia: a Detailed Bio-molecular and Mechanistic Landscape.

Contaminated rice is a major source of food poisoning in human communities where our earlier study showed Stenotrophomonas maltophilia, a Gram-negative bacillus, has been a major contaminant of the stored rice. In the present study, mono- and di-unsaturated fatty acids (UFAs) such as 18:1 ω 7 c, 16:1 ω 6 c, 16:1 ω 7 c, and 18:2 ω 6,9 c long-chain fatty acids have been found as the chief constituents of S. maltophilia boiled cell lysate. Throughout the study, both acute and chronic exposure of the cell lysate showed a decrease in the locomotor activity and a time-dependent increase of the depression (p < 0.001-0.0001, two-way ANOVA), supported by bioamine (dopamine, noradrenaline, adrenaline, serotonin, and GABA) depletion in rodents' brain possibly due to UFA-amino acid decarboxylase interaction favoring bioamine depletion as revealed by our study. Furthermore, the UFA-rich cell lysate revealed dose-dependent inhibition of murine brain microglial cell viability in vitro with concomitant increase of reactive oxygen species (ROS) inside the cell. Destruction of neuroprotective and neurotrophin releasing microglial cells, augmentation of brain ROS, and inflaming brain tissue resulting in infiltration of polymorphonuclear leucocytes also suggest to cause neurotoxicity by UFA derived from Stenotrophomonas maltophilia.

The Effect of Resveratrol on Sphingosine-1 and Oxidative/ Nitrosative Stress in an Experimental Heart Ischemia Reperfusion Model

Abstract Objectives: Resveratrol (RSV) is a natural polyphenolic compound showing significant antioxidant effects. In this study, we aimed to investigate the effects of resveratrol on the sphingosine-1-phosphate (S1P) and oxidative stress biomarkers in hearth ischemia-reperfusion (I/R). Materials and Methods: The biochemical and histopathological effects of RSV on cardiac ischemia-reperfusion injury were investigated through ELISA- and light microscope. Results: We observed statistically significant differences between the treatment group and the control group in terms of malondialdehyde (MDA) level, catalase (CAT) and superoxide dismutase (SOD) activities (p&lt;0.05). Histopathologically, we also observed decreased Polymorphonuclear Leucocyte (PMNL) infiltration, myocardial edema, miyositolysis in the treatment group compared to the I/R and sham groups. Conclusion: Resveratrol may play an important role in cardiac I/R injury through its anti-inflammatory and antioxidant effects which were biochemically and histopathologically confirmed in the present study.

Febuxostat treatment attenuates oxidative stress and inflammation due to ischemia-reperfusion injury through the necrotic pathway in skin flap of animal model

BackgroundIschemia-reperfusion (I/R) injury is a major contributor to skin flap necrosis, which is a serious complication of reconstructive surgery. The purpose of this study was to evaluate the protective effect of treatment with febuxostat, a selective xanthine oxidase inhibitor, on I/R injury in the skin flap of an animal (rat) model. MethodsSuperficial epigastric flaps were raised in Sprague-Dawley rats and subjected to ischemia for 3 h. Febuxostat at a dose of 10 mg/kg/day was administered to rats in drinking water from 1 week before the surgery (Feb group). Control animals received no drugs (Con group). The mean ratio of flap survival and contraction was evaluated and compared between animals with and without administration of febuxostat on day 5 after the surgery. In addition, infiltration by polymorphonuclear leukocytes and muscles of the panniculus carnosus in the flap were histologically evaluated using hematoxylin-eosin staining. Furthermore, xanthine oxidase activity, ATP levels, superoxide dismutase activity, and expression of 8-hydroxy-2′-deoxyguanosine (8-OHdG), tumor necrosis factor-α, and interleukin-1β were quantitatively assessed in the skin flap 24 h after the surgery. ResultsIn the Feb group, the survival and contraction rates at the 5 d timepoint post-surgery were significantly higher and lower than those in the Con group, respectively. Histological analysis showed significant reduction in polymorphonuclear leukocyte infiltration and muscle injury scores due to I/R injury in the Feb group. The expression of 8-OHdG was also significantly inhibited in animals administered febuxostat. Biochemical analysis showed a significant reduction in xanthine oxidase activity and significant increases in ATP levels and superoxide dismutase activity in the Feb group. Furthermore, the expression of interleukin-1β was significantly lower in the Feb group than in the Con group. ConclusionFebuxostat, which is clinically used for the treatment of hyperuricemia, was effective against necrosis of the skin flap via inhibition of oxidative stress and inflammation caused by I/R injury.

Oxidative and pro-inflammatory lung injury induced by desflurane inhalation in rats and the protective effect of rutin.

Desflurane is a mainstay of general inhaled anesthetics with a methyl ethyl ether structure and is widely used in clinical practice. It has been reported to induce inflammation and lipid peroxidation in rat pulmonary parenchyma, to increase alveolar macrophages, and to cause peribronchial infiltration and edema. Rutin, a flavonoid vitamin P1, is known to have biological properties including acting as an antioxidant, an anti-inflammatory, and an inhibitor of bronchoalveolar polymorphonuclear leukocyte (PNL) infiltration. The aim of this study is to examine the effects of rutin on desflurane-induced pulmonary injury using biochemical and histopathological methods. The rats were divided into 3 groups (n = 6 each): healthy control (HC), rutin+desflurane-treated (DRT) and desflurane-only (DSF). Briefly, 50 mg/kg of rutin was given orally to the DRT group and an equal volume of normal saline was given to the DSF and HC groups. After 1 h, anesthesia was induced and maintained in the DRT and DSF groups for 2 h. After the rats had been sacrificed, the lungs were removed. Malondialdehyde (MDA), total glutathione (GSH), tumor necrosis factor alpha (TNF-α), and nuclear factor kappa B (NF-κB) levels were measured in the excised lung tissue. The removed tissues were also fixed in 10% formalin, and the obtained sections were stained with hematoxylin and eosin (H&E) and evaluated under light microscopy. The biochemical and histopathological results of the DRT group were compared with those obtained from the DSF and HC groups. Desflurane increased MDA, TNF-α and NF-κB, and decreased GSH in lung tissue. The PNL infiltration, alveolar macrophages, hemorrhage, alveolar damage, and edema were observed in the lung tissue of the DSF group. Rutin was histopathologically shown to protect lung tissue from oxidative stress by preventing an increase in oxidant parameters and a decrease in antioxidants. The results suggest that rutin may be useful in the treatment of desflurane-associated lung injury.

The Effect of Adipose-Derived Stromal Vascular Fraction Cells to Abdominal Wall Fascia Defects in Rats: An Experimental Study

Objective Various synthetic and biological meshes have been developed to reduce recurrence and complications in ventral incisional hernia repairs. Adipose tissue is a rich reserve for mesenchymal stromal cells. In the present study we aimed to examine the effects of adipose-derived mesenchymal stromal cells (AD-MSCs) on abdominal incisional hernia repairs in rats. Materials and Methods The study involved 32 male Wistar-Albino rats, weighing 200–250 g, which were divided into three groups. In Group 1 (control group) only an incisional hernia model was created. In Group 2, the incisional hernia model was created and 1 ml stromal vascular fraction (SVF), obtained from inguinal lipectomy material and containing mesenchymal stromal cells, was injected into the edges of the defect in the same session. In Group 3, only the incisional hernia model was created in the first stage and after 14 days, 1 ml of SVF was injected into the edges of the defect. Skin incisions of rats in Group 1 and 2 were opened on postoperative day 28 while in group 3 were opened on day 42. Peritoneal formation in abdominal wall defect was evaluated macroscopically and histopathologically. Results Peritoneal formation was significantly superior in Groups 2 and 3 than in Group 1 (p: 0.031). In histopathological evaluation, the structural distortion and polymorphonuclear leukocyte (PMNL) levels were significantly higher in Group 1 than in Group 3 (p: 0.048 and p: 0.046, respectively). Granulation, capillary density, fibrosis and collagen organization were higher in Group 2 and 3, however this difference was not statistically significant (p > 0.05). Conclusions Adipose-derived stromal vascular fraction cells obtained from inguinal lipectomy material in rats positively affect the repair of abdominal incisional hernias by increasing peritoneal formation, and reducing structural distortion and PMNL infiltration.

Glycoproteins From Rabdosia japonica var. glaucocalyx Regulate Macrophage Polarization and Alleviate Lipopolysaccharide-Induced Acute Lung Injury in Mice via TLR4/NF-κB Pathway

Background and Aims: Rabdosia japonica var. glaucocalyx is a traditional Chinese medicine (TCM) for various inflammatory diseases. This present work aimed to investigate the protective effects of R. japonica var. glaucocalyx glycoproteins on lipopolysaccharide (LPS)-induced acute lung injury (ALI) and the potential mechanism. Methods: Glycoproteins (XPS) were isolated from R. japonica var. glaucocalyx, and homogeneous glycoprotein (XPS5-1) was purified from XPS. ANA-1 cells were used to observe the effect of glycoproteins on the secretion of inflammatory mediators by enzyme-linked immunosorbent assay (ELISA). Flow cytometry assay, immunofluorescence assay, and Western blot analysis were performed to detect macrophage polarization in vitro. The ALI model was induced by LPS via intratracheal instillation, and XPS (20, 40, and 80 mg/kg) was administered intragastrically 2 h later. The mechanisms of XPS against ALI were investigated by Western blot, ELISA, and immunohistochemistry. Results: In vitro, XPS and XPS5-1 downregulated LPS-induced proinflammatory mediators production including tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), IL-6, and nitric oxide (NO) and upregulated LPS-induced IL-10 secretion. The LPS-stimulated macrophage polarization was also modulated from M1 to M2. In vivo, XPS maintained pulmonary histology with significantly reducing protein concentration and numbers of mononuclear cells in bronchoalveolar lavage fluid (BALF). The level of IL-10 in BALF was upregulated by XPS treatment. The level of cytokines including TNF-α, IL-1β, and IL-6 was downregulated. XPS also decreased infiltration of macrophages and polymorphonuclear leukocytes (PMNs) in lung. XPS suppressed the expression of key proteins in the TLR4/NF-κB signal pathway. Conclusion: XPS was demonstrated to be a potential agent for treating ALI. Our findings might provide evidence supporting the traditional application of R. japonica var. glaucocalyx in inflammation-linked diseases.

The Effect of Ceruloplasmin Against Ischemia-Reperfusion Injury in Epigastric Island Flap in Rats

Flap surgery is frequently used in plastic surgery to close tissue defects. Ischemia-reperfusion (I/R) injury is a significant problem resulting in partial or total flap necrosis. This study aimed to investigate the effect of ceruloplasmin on I/R injury in epigastric island flaps in rats. A total of 32 male Sprague-Dawley rats were divided into four groups with eight rats in each group: The flap was not elevated in Group I; the flap was elevated without ischemia or any application in Group II, after the intraperitoneal saline and ceruloplasmin application the flaps were elevated and ischemia was created in group III-IV, respectively. Bilateral epigastric artery flap was elevated in all groups except Group I. After 6 h of ischemia, the flap was reperfused and inset. Samples were taken from the right and left side of the flap area in other groups at the postoperative 24th h for biochemical analysis (catalase and malondialdehyde-MDA) and the seventh postoperative day for histopathological analysis (Modified Verhofstad score and epidermal thicknesses), respectively. Image analysis for necrosis areas was performed on photos taken on the 7th d. Catalase level was significantly higher in Group IV.(0.15 ± 0.04 U/mg protein) (P < 0.05) Necrosis area percentage(14.4% ± 3.3%),MDA(3.6 ± 0.9 nmol/mg protein), edema(3), necrosis(2.75), and polymorphonuclear leukocyte infiltration(2.87) scores were significantly higher in group III.(P < 0.05). Fibroblast proliferation, collagen density (0.25), vascular density (0.25) scores and epidermal thickness (15.68 µm,) was significantly lower in group III. (P < 0.05) CONCLUSIONS: Our study demonstrated that ceruloplasmin application before ischemia reduced I/R injury in epigastric island flaps in rats.

Effects of orally administered Euglena gracilis and its reserve polysaccharide, paramylon, on gastric dysplasia in A4gnt knockout mice

Euglena gracilis is widely utilized as food or supplement to promote human and animal health, as it contains rich nutrients. In this study, we administered spray-dried powder of E. gracilis and paramylon, β-glucan stored in E. gracilis cells, to A4gnt knockout (KO) mice. A4gnt KO mice are a mutant mouse model that spontaneously develops gastric cancer through hyperplasia-dysplasia-adenocarcinoma sequence in the antrum of the stomach, and we observed the effects of E. gracilis and paramylon on the early involvements of A4gnt KO mice. Male and female 10-week-old A4gnt KO mice and their age-matched wildtype C57BL/6J mice were orally administered with 50 mg of E. gracilis or paramylon suspended in saline or saline as a control. After 3-week administration, animals were euthanatized and the stomach was examined histopathologically and immunohistochemically. Gene expression patterns of the stomach, which have been reported to be altered with A4gnt KO, and IgA concentration in small intestine were also analyzed with real-time PCR and ELISA, respectively. Administration of Euglena significantly reduced the number of stimulated CD3-positive T-lymphocytes in pyloric mucosa of A4gnt KO mice and tend to reduce polymorphonuclear leukocytes infiltration. Euglena administration further downregulated the expression of Il11 and Cxcl1 of A4gnt KO mice. Euglena administration also affected IgA concentration in small intestinal contents of A4gnt KO mice. Paramylon administration reduced the number of CD3-positive lymphocytes in pyloric mucosa of A4gnt KO mice, and downregulated the expressions of Il11 and Ccl2 of A4gnt KO mice. Although we found no significant effects on gross and microscopic signs of gastric dysplasia and cell proliferation, the present study suggests that the administration of Euglena and paramylon may ameliorate the early involvements of A4gnt mice through the effects on inflammatory reactions in the gastric mucosa. The cancer-preventing effects should be studied with long-term experiments until actual gastric cancer formation.

Local shifts in inflammatory and resolving lipid mediators in response to tendon overuse.

Tendon inflammation has been implicated in both adaptive connective tissue remodeling and overuse-induced tendinopathy. Lipid mediators control both the initiation and resolution of inflammation, but their roles within tendon are largely unknown. Here, we profiled local shifts in intratendinous lipid mediators via liquid chromatography-tandem mass spectrometry in response to synergist ablation-induced plantaris tendon overuse. Sixty-four individual lipid mediators were detected in homogenates of plantaris tendons from ambulatory control rats. This included many bioactive metabolites of the cyclooxygenase (COX), lipoxygenase (LOX), and epoxygenase (CYP) pathways. Synergist ablation induced a robust inflammatory response at day 3 post-surgery characterized by epitenon infiltration of polymorphonuclear leukocytes and monocytes/macrophages (MΦ), heightened expression of inflammation-related genes, and increased intratendinous concentrations of the pro-inflammatory eicosanoids thromboxane B2 and prostaglandin E2 . By day 7, MΦ became the predominant myeloid cell type in tendon and there were further delayed increases in other COX metabolites including prostaglandins D2 , F2α , and I2 . Specialized pro-resolving mediators including protectin D1, resolvin D2 and D6, as well as related pathway markers of D-resolvins (17-hydroxy-docosahexaenoic acid), E-resolvins (18-hydroxy-eicosapentaenoic acid), and lipoxins (15-hydroxy-eicosatetraenoic acid) were also increased locally in response to tendon overuse, as were anti-inflammatory fatty acid epoxides of the CYP pathway (eg, epoxy-eicosatrienoic acids). Nevertheless, intratendinous prostaglandins remained markedly increased even following 28 days of tendon overuse together with a lingering MΦ presence. These data reveal a delayed and prolonged local inflammatory response to tendon overuse characterized by an overwhelming predominance of pro-inflammatory eicosanoids and a relative lack of specialized pro-resolving lipid mediators.

Increased hepatic JNK activation by ethanol is mediated by transcriptional suppression of mitogen‐activated protein kinase phosphatase 1 (Mkp1): role of cAMP‐specific protein kinase A

Background Increased and sustained JNK activation has been shown to be detrimental for hepatocyte survival and liver injury in various diseases, including alcohol-associated liver disease (ALD). Among three JNK isoforms, JNK1 and 2 are expressed in hepatocytes and both play a role in mouse models of ALD. Mkp1 phosphatase has been identified as a regulator of JNK activation. A previous study showed that Mkp1 degradation plays a critical role in sustained JNK activation and hepatocyte death from acute ethanol exposure. However, the effect of chronic ethanol exposure on hepatocyte Mkp1 expression is not known. Methods Male C57B/6 mice were subjected to intragastric (iG) feeding of ethanol for 8 weeks with weekly ethanol binge (5g/kg) starting in the second week of iG feeding. Pair-fed control mice were fed an isocaloric high-fat liquid diet. Plasma ALT, AST and histologic grading were performed to evaluate steatosis, liver inflammation and injury. Liver immunohistochemistry was performed to evaluate apoptosis (TUNEL staining). Myeloperoxidase (MPO) activity assay and chloroacetate esterase (CAE) staining for neutrophil infiltration were also performed. Liver RNA sequencing was performed on an Illumina NextSeq 500. Data analysis was done using the Tuxedo suite pipeline. Validation studies were carried out by performing real time qPCR and Western blot. Primary mouse hepatocytes were exposed to 50 mM ethanol for 48 hours followed by TNF stimulation. Statistical analysis was performed using ANOVA. Results Histological examination of liver sections confirmed the development of severe alcohol-associated steatohepatitis in ethanol fed mice via significant micro- and macro-vesicular steatosis, polymorphonuclear leukocyte (PMN) infiltration, CAE and TUNEL-positive cells. ALT and AST levels were significantly elevated in ethanol-fed mice. JNK activation was evident by Western blot analysis of phosphorylated JNK levels. RNA Seq analysis showed significant changes in expression of several Mkps; however, only Mkp1 levels were significantly downregulated, which was further validated by real time qPCR. Importantly, ethanol exposure of primary mouse hepatocytes led to a significant decrease in Mkp1 mRNA expression which further declined following TNF stimulation. These changes in Mkp1 were accompanied by sustained JNK activation and cell death. Moreover, ethanol exposure and TNF stimulation markedly increased cAMP specific PDE4 expression resulting in reduction in hepatocyte cAMP levels. Significantly, PDE4 inhibition and PKA specific agonists prevented the suppressive effect of ethanol and TNF on Mkp1 and resultant activation of JNK and cell death. Conclusion Our results strongly implicate ethanol-induced decline in hepatic Mkp1 as a significant mechanism regulating JNK activation and hepatocyte death in ALD. Further, these data strongly suggest that the ethanol induced decrease in cAMP/PKA signaling plays a causal role in regulating the Mkp1/JNK axis in hepatocytes and development of liver injury.

IL-36α Exerts Proinflammatory Effects in Aspergillus fumigatus Keratitis of Mice Through the Pathway of IL-36α/IL-36R/NF-κB.

PurposeTo explore the role of IL-36α in corneas infected by Aspergillus fumigatus.MethodsThe experimental group was comprised of 15 corneas with fungal keratitis, and 15 healthy donor corneas were included in the control group. IL-36α was detected in normal and infected corneas of humans and C57BL/6 mice. Mice corneas were infected with A. fumigatus with or without pretreatment of recombinant mouse (rm) IL-36α and IL-36α neutralizing antibody (Ab). Primary macrophages were stimulated with 75% ethanol-killed A. fumigatus with or without pretreatment of rmIL-36α. The severity of the disease was documented by clinical score and photographs with a slit lamp. PCR, western blot, and immunostaining were used to determine the expression of IL-36α, IL-1β, IL-6, and TNF-α. Polymorphonuclear neutrophilic leukocyte infiltration was assessed by myeloperoxidase (MPO) assay and flow cytometry. Macrophage infiltration was tested by immunofluorescent staining and flow cytometry.ResultsIL-36α mRNA and protein were significantly elevated in human and mice corneas after infection. The rmIL-36α treatment of C57BL/6 mice increased clinical score, MPO levels, macrophage infiltration, and expression of the proinflammatory cytokines IL-1β, IL-6, and TNF-α compared with the infected controls, which showed a decrease due to IL-36α Ab treatment. In primary macrophages, IL-36α expression was also significantly increased by A. fumigatus. The rmIL-36α treatment upregulated IL-1β, IL-6, and phosphorylated nuclear factor (NF)-κB expression, which was significantly inhibited by rmIL-36Ra.ConclusionsIL-36α act as a proinflammatory cytokine in A. fumigatus keratitis by promoting the infiltration of neutrophils and macrophages and increasing the secretion of IL-1β, IL-6, and TNF-α, in addition to regulating expression of phosphorylated NF-κB.

The Effect of Botulinum Toxin on Flap Viability of the Posterior Thigh Perforator Flap in Rats

BackgroundThe use of perforator propeller flaps in lower limb reconstruction has increased recently. Many pharmacological agents are used to increase flap viability. Botulinum toxin has been used in various types of flaps in the literature. However, there is no study regarding the use of botulinum toxin in the lower limb propeller flaps. This study investigates the effect of botulinum toxin administration on flap survival for lower limb propeller flap in rats. Materials and methodsThe study included 20 male Wistar albino rats, divided into two groups with a flap rotation of 90° in group 1 and 180° in group 2. In both groups, botulinum toxin was administered to the right thigh and a physiological saline solution was applied to the left thigh. Five days later, flaps were elevated over the posterior aspect of the right and left thighs and inset after 90° and 180° rotation was performed. Histopathological, immunohistochemical, and necrosis area analyses were performed. ResultsNecrosis area, edema, polymorphonuclear leukocyte infiltration, and necrosis were found to be higher on the left side of the groups, whereas epidermal thickness, collagen density, vascularization, and hair root density were found to be higher on the right side of the groups. No significant difference was found between the right posterior thighs in either group on any parameter other than vascularization. Histopathologically and immunochemically statistically significant differences were found between the two groups. ConclusionsThe present study found that botulinum toxin increases flap viability in lower limb perforator-based propeller flaps.

Dietary supplementation with fructooligosaccharides ameliorates allergy development following DEHP exposure in mice

ABSTRACTFructooligosaccharides (FOS), a prebiotic supplement, can enhance immunological responses, probably through regulation of gastrointestinal microflora. On the other hand, some environmental pollutants have adjuvanticity against allergic reactions/diseases. The purpose of this study was to evaluate the effects of dietary supplementation with FOS on a murine model of enhanced ovalbumin (OVA)-related allergic peritonitis induced by di-(2-ethylhexyl) phthalate (DEHP). C3H/HeN mice were intraperitoneally administered OVA and DEHP, and fed a diet containing 2.5% FOS. Thereafter, several parameters were evaluated. Supplementation with FOS alleviated OVA plus DEHP-related peritoneal inflammation characterized by infiltration of polymorphonuclear leukocytes including neutrophils in the peritoneal cavity. Also, FOS administration declined the number of mast cells of OVA + DEHP treated mice. FOS suppressed the elevated protein level of interleukin-5, eotaxin, and keratinocyte-derived chemoattractant, in the peritoneal lavage fluids. Our results suggest that dietary supplementation with FOS can prevent or ameliorate enhanced allergic peritoneal inflammation induced by DEHP.

Effect of Single and Repeated Administration of Amitriptyline on Experimental Gastric Ulcer

Amitriptyline is an antidepressant drug widely used as analgesic in the management of chronic pain syndromes. It is also attracting the attention of researchers as an alternative agent for peptic ulcer management because studies have shown that most patients with peptic ulcer also suffer from depression. Thus, this study was conducted to investigate gastroprotective effects of single and repeated dose administration of amitriptyline against indomethacin-induced gastric injury in rats. Rats were randomly divided into three groups of eight rats/each. For 14 days, Group I (control) and Group II (single dose) received distilled water; Group III (repeated dose) received 10 mg/kg amitriptyline orally once daily. On day 15, Group I was given distilled water; Group II and III were given 10 mg/kg amitriptyline orally. Thirty minutes later, indomethacin (25 mg/kg) was administrated orally to all the groups. Six hours after indomethacin administration, animals were sacrificed under thiopental sodium anaesthesia, stomachs were rapidly removed, cut along the greater curvature and cleaned. The stomachs were macroscopically evaluated; total area of stomach and visible ulcerated areas were measured using millimeter square paper. Finally, the stomachs were fixed in 10% formalin for histopathologic valuation. Single dose administration of amitriptyline significantly reduced ulcer indexes and gastric erosions as compared to the control group.However, its role against inflammatory reactions such as necrosis, polymorphonuclear leucocyte infiltration, neutrophil and eosinophil reactions were statistically insignificant. On the other hand, repeated dose administration of amitriptilyine produced significant gastroprotective effects as observed from macroscopical and histopathological evaluation results. Moreover, there was also a significant difference in the gastroprotective effects between single and repeated dose of amitriptyline against indomethacin-induced gastric damage. In conclusion, the results of this study evealed that repeated dose administration of amitriptyline showed better gastroprotective effects compared to single dose amitriptyline administration and the control group.&#x0D; Keywords: amitriptyline, gastroprotective effects, indomethacin, histopathologic evaluation, rats

Mouse model for endometriosis is characterized by proliferation and inflammation but not epithelial-to-mesenchymal transition and fibrosis

Endometriosis is a common disorder of unknown etiology, and non-surgical therapies are still a challenge. To understand the pathogenesis and preclinical testing of drugs for endometriosis, animal models are highly desirous. Herein, we carried out longitudinal characterization of a mouse model for endometriosis where uterine tissue was transplanted onto the intestinal mesentery. During the course of lesion development from day 15 to 60 post-induction, the ectopic endometrium became pale, fluid-filled and the animals developed peritoneal adhesions. Most lesions resembled a well-differentiated type of endometriosis and ~13% of animals had mixed type of lesions. There was extensive stromal compaction in the ectopic tissue. During the progression of endometriosis, there was increased proliferation of epithelial and stromal cells as evident by PCNA staining. Cyp19a1 (aromatase) mRNA was detected in the ectopic lesions on day 15 and 30 post-induction of endometriosis, by day 60 the expression was reduced. As compared to the control endometrium, the mRNA levels of Esr1 progressively reduced while the levels of inflammation associated genes (Esr2, Ifng, Tnf and Il1b) increased in the ectopic lesions. Infiltration of macrophages and polymorphonuclear leucocytes was also observed in the ectopic lesions indicative of inflammation. As compared to control, there was no change in levels of Cytokeratin and E-cadherin in the epithelial cells of ectopic endometrium. We did not observe excessive collagen deposition or α -SMA positive myofibroblasts in the stroma of the ectopic endometrium. Thus, epithelial-to-mesenchymal transition and fibrosis are not detected in the mouse model of endometriosis. Our results show that the mouse model of endometriosis mimics some but not all the features of human endometriosis.

Antioxidant effect of acetyl-l-carnitine against cisplatin-induced cardiotoxicity.

ObjectiveCisplatin (CDDP) toxicity is a dose-limiting clinical problem in clinical practice, mainly because of nephrotoxicity or ototoxicity. However, the mechanism of CDDP-induced cardiotoxicity is poorly understood. Acetyl-l-carnitine (ALCAR) is an antioxidant agent with protective effects against the side effects of various chemotherapeutics. CDDP-induced cardiotoxicity and the protective role of ALCAR were evaluated in this study.MethodsMorphological changes were evaluated in hematoxylin and eosin-stained sections, and immunohistochemistry for caspase-3, superoxide dismutase-2 (SOD-2), inducible nitrite oxide synthase (iNOS), cyclooxygenase-2, and Bcl-2 was performed using the hearts of athymic nude mice carrying xenograft neuroblastoma tumors. Mice were randomized (six/group) to the control, CDDP (16 mg/kg), and ALCAR (200 mg/kg)+CDDP (16 mg/kg) groups. Results were analyzed using nonparametric tests.ResultsNo difference was observed in the rates of cardiac necrosis, dilated/congested blood vessels, hemorrhage, polymorphonuclear leukocyte infiltration, edema, and pyknotic nuclei among the groups. SOD-2 expression was increased in the CDDP group but not in the ALCAR+CDDP group. iNOS, Bcl-2, and caspase-3 levels were not significantly different among the groups.ConclusionsALCAR might be a candidate protective agent for CDDP-induced cardiotoxicity. SOD-2, as a member of the oxidant system, should be evaluated in further studies as a biomarker of cardiotoxicity.

Brief topical and intraluminal use of Carolina rinse solution does not attenuate experimental ischemia and reperfusion injury in rabbit jejunum

ABSTRACT Fifteen New Zealand adult rabbits were randomly allocated into three groups: Sham-operated (group A), Ischemia and Reperfusion (group B) and Carolina Rinse Solution (CRS) (group C). Groups B and C were subjected to one hour of ischemia and two hours of reperfusion. In group C, ten minutes before reperfusion, the bowel lumen was filled with CRS, and the segment immersed in CRS. Necrosis and loss of integrity of the villi were visible in groups B and C. Edema of the submucosa and circular muscle was observed in all groups. Hemorrhage was observed in different layers for groups B and C, but group C showed more severe hemorrhage in different layers during reperfusion. All groups showed polymorphonuclear leukocyte infiltration on the base of the mucosa, submucosa, and longitudinal muscle, in addition to polymorphonuclear leukocytes margination in the mucosal and submucosal vessels. Necrosis of enterocytes, muscles, crypts of Lieberkühn and myenteric plexus was observed in groups B and C during reperfusion. Topical and intraluminal Carolina Rinse Solution did not attenuate the effects of ischemia and reperfusion in the small intestine of rabbits.

Decreased neutrophil levels in mice with traumatic brain injury after cape administration

IntroductionPeripheral leukocytes can worsen brain damage due to the release of cytotoxic mediators that interfere the blood brain barrier function. One of the oxidants released by activation leukocyte is hypochlorite acid (HOCl) which is formed through the myeloperoxidase (MPO)-H2O2–Cl- system. The neuroprotective effects of an experimental anti-inflammatory drug Caffeic Acid Phenethyl Ester (CAPE) tested in a Traumatic brain injury (TBI) model using Myeloperoxidase (MPO) analysis. MethodsThis study compares the acute inflammatory response to TBI over time, as measured by MPO activity. Adult Sprague mice were treated for head trauma with marmarou model. At 24 h before trauma, all animals were blood test (n = 10) to examine MPO, then the animal was divided into 2 groups of injured animals treated with CAPE (n = 5), and those not treated with CAPE (n = 5). We used the MPO test to identify the level of polymorphonuclear leukocytes (PMNL) on day 4 and day 7. ResultsShowed an increase in PMNL infiltration in CAPE untreated animals, this change significantly (P < 0.05) decreased at group of animals treated with CAPE. MPO serum activity in the CAPE untreated group vs treated with CAPE on day 4 ± 11920410.076 (Standard deviation {SD} 895355.169) vs 6663184.485 (SD 895355.169) p < 0,05 and on day 7 ± 14223286.992 (SD 802762.401) vs 9284222.028 (SD 953098.093) p < 0,05. These data indicate that MPO activity after TBI increases on day 4 also on day 7 and improves after being treated with CAPE. ConclusionCAPE can reduce Neutrophil serum levels there by preventing brain damage in TBI.