Morphological Characteristics of the Results of Experimental Modeling of Septic Peritonitis

SUMMARY: Experimental studies devoted to the study of the mechanisms of the pathogenesis of acute peritonitis and the development of new methods of medical and surgical treatment are becoming increasingly relevant. Today, experimental medicine knows many different ways to modeling septic peritonitis and eliminate it, but the role of the local immune system is underestimated, whereas it takes a direct part in inflammation. The objective of our work to study morphological features of results of experimental modeling of septic peritonitis in white rats. The study included 15 sexually mature white male rats weighing 276.75±6.56 grams. A simulation of septic peritonitis was performed by perforating the upper part of the cecum with four punctures with a G16 injection needle. As a result of the experiment, after examination of the peritoneal cavity, all 15 animals were diagnosed with omentum tamponade of perforated damage to the caecum. In 11 cases, the perforated wall of the caecum was covered by the greater omentum (73.34 %), and in the other 4 animals, tamponade was performed by one of the epididymal omentum (26.66 %). The initial stage of tamponade with the greater or epididymal omentums of a perforated caecum begins on the first day of the experiment and consists of tight interstitial consolidation between them, as well as in the invasion of blood vessels from the omentum side to the focus of infection, which ensure the delivery of the appropriate immunocompetent cells. As a result of this process, intensive lymphoid infiltrates are formed in this area, as well as the growth of adipose tissue, which isolates the inflammatory focus from the peritoneal cavity with a thick layer.

Las investigaciones experimentales dedicadas al estudio de los mecanismos de patogénesis de la peritonitis aguda y el desarrollo de nuevos métodos de tratamiento médico y quirúrgico son cada vez más relevantes. Hoy en día, la medicina experimental conoce muchas formas diferentes de modelar la peritonitis séptica y eliminarla, pero se subestima el papel del sistema inmunológico local, mientras que él participa directamente en la inflamación. El objetivo de nuestro trabajo fue estudiar las características morfológicas de los resultados del modelado experimental de peritonitis séptica en ratas blancas. El estudio incluyó 15 ratas macho blancas, sexualmente maduras que pesaban 276,75 ± 6,56 gramos. Se realizó una simulación de peritonitis séptica perforando la parte superior del ciego con cuatro punciones con una aguja de inyección G16. Como resultado del experimento, después del examen de la cavidad peritoneal, a los 15 animales se les diagnosticó taponamiento del omento o lesión perforada del ciego. En 11 casos, la pared perforada del ciego fue recubierta por el omento mayor (73,34 %), y en los otros 4 animales el taponamiento se realizó por uno de los epidídimos (26,66 %). La etapa inicial del taponamiento con omento mayor o epidídimo de un ciego perforado comienza el primer día del experimento y consiste en una estrecha consolidación intersticial entre ellos, así como en la invasión de los vasos sanguíneos desde el lado del omento hasta el foco de infección, que aseguran la entrega de las células inmunocompetentes apropiadas. Como resultado de este proceso, se forman intensos infiltrados linfoides en esta zona, así como el crecimiento de tejido adiposo, que aísla el foco inflamatorio de la cavidad peritoneal con una gruesa capa.

Effects of Formyl Peptide Receptor Agonists Ac9-12 and WKYMV in In Vivo and In Vitro Acute Inflammatory Experimental Models.

Formyl peptide receptors (Fprs) are a G-protein-coupled receptor family mainly expressed on leukocytes. The activation of Fpr1 and Fpr2 triggers a cascade of signaling events, leading to leukocyte migration, cytokine release, and increased phagocytosis. In this study, we evaluate the effects of the Fpr1 and Fpr2 agonists Ac9-12 and WKYMV, respectively, in carrageenan-induced acute peritonitis and LPS-stimulated macrophages. Peritonitis was induced in male C57BL/6 mice through the intraperitoneal injection of 1 mL of 3% carrageenan solution or saline (control). Pre-treatments with Ac9-12 and WKYMV reduced leukocyte influx to the peritoneal cavity, particularly neutrophils and monocytes, and the release of IL-1ß. The addition of the Fpr2 antagonist WRW4 reversed only the anti-inflammatory actions of WKYMV. In vitro, the administration of Boc2 and WRW4 reversed the effects of Ac9-12 and WKYMV, respectively, in the production of IL-6 by LPS-stimulated macrophages. These biological effects of peptides were differently regulated by ERK and p38 signaling pathways. Lipidomic analysis evidenced that Ac9-12 and WKYMV altered the intracellular lipid profile of LPS-stimulated macrophages, revealing an increased concentration of several glycerophospholipids, suggesting regulation of inflammatory pathways triggered by LPS. Overall, our data indicate the therapeutic potential of Ac9-12 and WKYMV via Fpr1 or Fpr2-activation in the inflammatory response and macrophage activation.

Vatairea guianensis lectin stimulates changes in gene expression and release of TNF-α from rat peritoneal macrophages via glycoconjugate binding.

Using a rat model of peritonitis, we herein report the inflammatory effect induced by the lectin isolated from Vatairea guianensis (VGL) seeds in the context of interactions between VGL and both toll-like receptor 4 (TLR4) and tumor necrosis factor receptor 1 (TNFR1). Peritoneal macrophages were stimulated with VGL for dose-dependent gene expression and release of TNF-α. In vivo results showed that VGL (1 mg/kg; intraperitoneal) induced peritonitis in female Wistar rats. Leukocyte migration, macrophage activation, and protein leakage were measured 3 and 6 hours after induction. In vitro, peritoneal macrophages were stimulated with VGL for gene expression and TNF-α dosage (mean ± SEM (n = 6), analysis of variance, and Bonferroni's test (P < .05)). In silico, VGL structure was applied in molecular docking with representative glycans. It was found that (a) VGL increases vascular permeability and stimulates leukocyte migration, both rolling and adhesion; (b) lectin-induced neutrophil migration occurs via macrophage stimulation, both in vitro and in vivo; (c) lectin interacts with TLR4 and TNFR1; and (d) stimulates TNF-α gene expression (RT-PCR) and release from peritoneal macrophages. Thus, upon lectin-glycan binding on the cell surface, our results suggest that VGL induces an acute inflammatory response, in turn activating the release of peritoneal macrophages via TNF-α and TLR and/or TNFR receptor pathways.

Polysaccaride-rich extract of Caesalpina ferrea stem barks attenuates mice acute inflammation induced by zymosan: Oxidative stress modulation.

ETHNOPHARMACOLOGICAL RELEVANCE: Stem barks of Caesalpinia ferrea Mart. Ex Tul. (Caesalpiniaceae), also known as pau-ferro jucá or jucaína, are popularly used to treat contusions, diabetes, rheumatism and other inflammatory conditions in the form of tea, lick or decoction. OBJECTIVE: To evaluate the effect of the polysaccharide-rich extract obtained from C. ferrea stem barks (PE-Cf) in mice models of acute inflammation induced by zymosan and the involvement of oxidative stress biomarkers. MATERIALS AND METHODS: Mice were treated with PE-Cf (0.001, 0.01, 0.1, 1 mg/kg) by endovenous route (i.v.) or per oral (p.o.) 30 or 60 min before injection of the inflammatory stimuli zymosan (0.5 mg; intraperitoneal or subcutaneous intraplantar). The inflammatory parameters (edema, nociception, leukocyte migration) and oxidative stress markers (myeloperoxidase-MPO, malondialdehyde-MDA, nitrite, reduced glutathione-GSH, glutathione peroxidase-GPx) were evaluated in the models of paw edema (hidropletysmometry/expressed as ml or area under curve-AUC) and peritonitis (optical microscopy/expressed as n° of cells/mm3 of peritoneal fluid). Statistical analysis was performed by ANOVA, followed by Bonferroni test. RESULTS: PE-Cf (0.1, 0.01 and 1 mg/kg) dose-dependently inhibited paw edema, showing maximal effect (74%) at 1 mg/kg in the 5th (52 ± 9.6 µl vs. zymosan: 204 ± 3.6 µl). PE-Cf (1 mg/kg) also inhibited by 43% MPO activity in the paw tissues (17 ± 1 vs. zymosan: 30 ± 2.6 U/mg). Besides, 4 h after peritonitis induction, PE-Cf (1 mg/kg) reduced neutrophil migration by 84% (432 ± 45 vs. zymosan: 2651 ± 643 cells/mm3); visceral nociception by 76% (3 ± 0.6 vs. zymosan: 16 ± 4 writhes); nitric oxide by 73% (0.131 ± 0.033 vs. zymosan: 0.578 ± 0.185 NO2-/NO3-ml); MDA (98 ± 10 vs. zymosan:156 ± 21 U/ml), and increased GSH by 65% (736 ± 65 vs. zymosan: 259 ± 58 µmol/ml) and GPx by 72% (0.037 ± 0.007 vs. zymosan: 0.010 ± 0.005 U/mg protein). CONCLUSION: The polysaccharide-rich extract of Caesalpinia ferrea stem barks present anti-inflammatory and antioxidant effects in mice models of acute inflammation induced by zymosan.

MSU Crystals induce sterile IL-1ß secretion via P2X7 receptor activation and HMGB1 release.

BACKGROUD: The mechanism by which monosodium urate (MSU) crystals induce inflammation is not completely understood. Few studies have shown that MSU is capable of stimulating the release of IL-1ß in the absence of LPS treatment. The purinergic P2X7 receptor is involved in the release of IL-1ß in inflammatory settings caused by crystals, as is the case in silicosis. METHODS: We investigated the role of P2X7 receptor in sterile MSU-induced inflammation by evaluating peritonitis and paw edema. In in vitro models, we performed the experiments using peritoneal macrophages and THP-1 cells. We measured inflammatory parameters using ELISA and immunoblotting. We measured cell recruitment using cell phenotypic identification and hemocytometer counts. RESULTS: Our in vivo data showed that animals without P2X7 receptors generated less paw edema, less cell recruitment, and lower levels of IL-1ß release in a peritonitis model. In the in vitro model, we observed that MSU induced dye uptake by the P2X7 receptor. In the absence of the receptor, or when it was blocked, MSU crystals induced less IL-1ß release and this effect corresponded to the concentration of extracellular ATP. Moreover, MSU treatment induced HMGB1 release; pre-treatment with P2X7 antagonist reduced the amount of HMGB1 in cell supernatants. CONCLUSIONS: IL-1ß secretion induced by MSU depends on P2X7 receptor activation and involves HMGB1 release. GENERAL SIGNIFICANCE: We propose that cell activation caused by MSU crystals induces peritoneal macrophages and THP-1 cells to release ATP and HMGB1, causing IL-1ß secretion via P2X7 receptor activation.

Hernia de Amyand tipo 3. Reporte de caso / Amyand hernia type 3. Case report

Se describe a la hernia de Amyand como la presencia del apéndice cecal dentro de un saco herniario inguinal con un proceso inflamatorio-infeccioso o no, cuya frecuencia <1% de los casos de hernias inguinales. Exponer caso clínico y revisión bibliográfica de la hernia de Amyand tipo 3. Paciente masculino de 65 años de edad con enfermedad actual de inicio 10 días previo a su ingreso cuando posterior a esfuerzo físico presenta aumento de volumen no reductible en región inguinal derecha dolorosa, posteriormente 24 horas previo a su ingreso presenta dolor de fuerte intensidad en región inguinal derecha, persistencia del aumento de volumen y signos de flogosis, náuseas y alzas térmicas no cuantificadas, por lo cual acude a nuestra institución. Examen fisico: Abdomen blando depresible, doloroso a la palpación profunda en fosa ilíaca derecha, sin signos de irritación peritoneal, se apreciaba aumento de volumen en region inguinal derecha, no reductible, doloroso a la palpación con rubor y calor local, genitales masculinos con dolor a la palpación del testìculo derecho. Hernia inguinal derecha atascada. Intervención quirúrgica: Hernioplastia inguinal por técnica de cierre preperitoneal, sin colocación de malla, apendicectomía atípica por técnica de Pouchet. La hernia de Amyand es una patología poco frecuente, que se presenta <1% y acompañada de complicaciones <0,1%, debe sospecharse en pacientes que presente una patología herniaria derecha con leucocitosis y neutrofilia sin otro foco infeccioso demostrado, donde el diagnóstico principalmente se hace intraoperatorio como lo expone el presente caso(AU)

Amyand's hernia is described as the presence of the cecal appendix within an inguinal hernial sac with an inflammatory-infectious process or not, the frequency of which is <1% of cases of inguinal hernias. To present a clinical case and a bibliographic review of Amyand hernia type 3. A 65-year-old male patient with current disease that started 10 days before admission when, after physical effort, he presented a non-reducible increase in volume in the right inguinal region. Painful, later 24 hours before admission, he presented pain of strong intensity in the right inguinal region, persistence of increased volume and signs of phlogosis, nausea and thermal increases not quantified, for which he came to our institution. Physical exam: painful depressible soft abdomen on deep palpation in the right iliac fossa without signs of peritoneal irritation, volume increase was observed in the right inguinal region, not reducible, painful on palpation with flushing and local heat, male genitalia with pain on palpation of the right testicle. Stuck right inguinal hernia. Surgical intervention: Inguinal hernioplasty by preperitoneal closure technique without mesh placement, atypical appendectomy by Pouchet technique. Amyand's hernia is a rare pathology, which presents <1% and accompanied by complications <0.1%, it should be suspected in patients presenting a right hernia pathology with leukocytosis and neutrophilia without another proven infectious focus, where the diagnosis is mainly made intraoperatively as exposed in the present case(AU)

Peritonitis esclerosante primaria y secundaria: estudio comparativo / Primary and secondary sclerosing peritonitis: comparative study

Resumen Introducción: Existen dos tipos de peritonitis esclerosante (PE): primaria o idiopática y secundaria, generalmente a diálisis peritoneal (DP), y con menor frecuencia a otras patologías abdominales o sistémicas. Su mortalidad es alta. Objetivo: Comparar las características clínicas, estudios diagnósticos y tratamiento de pacientes con Peritonitis Esclerosante Primaria y Secundaria, definir si existen diferencias y determinar los principales elementos clínicos e imagenológicos que permitan hacer un diagnóstico precoz y mejorar los resultados terapéuticos. Material y Métodos: Se analizan 18 casos de PE diagnosticados en nuestro hospital, entre los años 2001-2014. Incluye una serie retrospectiva de 15 casos de PE secundaria (13 por diálisis peritoneal y 2 por cirrosis hepática). Se compara con un estudio prospectivo que incluye 3 pacientes con PE primaria. Resultados: Las principales diferencias se evidencian en la presentación clínica: PE primaria: se presenta con cuadro de obstrucción intestinal y baja de peso de distinta magnitud. PE secundaria: predominan el dolor abdominal, peritonitis recurrente y la falla de ultrafiltración. La tomografía computada de abdomen es útil, sobre todo cuando hay obstrucción intestinal. Ha hecho posible el diagnóstico preoperatorio. Conclusiones: Se requiere un alto índice de sospecha para el diagnóstico precoz de PE, sobre todo para la forma primaria. Debe sospecharse en todo paciente con dolor abdominal, vómitos recurrentes y baja de peso de cualquier magnitud; y en aquellos en diálisis peritoneal durante 5 años o más, que presenten dolor abdominal y/o peritonitis recurrente y/o falla de ultrafiltración.

Introduction: There are two types of sclerosing peritonitis (SP): primary or idiopathic and secondary, generally to peritoneal dialysis, and less frequently, to other abdominal or systemic pathologies. Mortality related to this is high. Objective: To compare the clinical feature, diagnostic studies and treatment of patients with Primary and Secondary Sclerosing Peritonitis, to define whether there are any differences and to establish the main clinical and imaging elements allowing for an early diagnosis and improving the therapeutic results. Material and Methods: An analysis of 18 SP cases diagnosed at our hospital between 2001-2014 was carried out. This includes a retrospective series of 15 cases of secondary SP (13 to peritoneal dialysis and 2 to liver cirrhosis). This is compared against a prospective study that includes 3 patients with primary SP. Results: The main differences became evident in the clinical presentation: Primary SP: occurs in an intestinal obstruction and a loss of weight scenario of varying degrees. Secondary SP: abdominal pain and recurrent peritonitis as well as ultrafiltration failure prevail. CT of the abdomen has proven to be useful, in particular in those cases where there is intestinal obstruction. It has made preoperative diagnostic possible. Conclusions: A high degree of suspicion is required for an SP early diagnosis, especially for the primary form. All patients presenting abdominal pain, recurrent vomiting and any degree of weight loss and those with five or more years of peritoneal dialysis presenting abdominal pain and/or recurrent peritonitis and/or ultrafiltration failure should raise a diagnosis suspicion.

2-Allylphenol Reduces IL-1ß and TNF-α, Promoting Antinociception through Adenosinergic, Anti-Inflammatory, and Antioxidant Mechanisms.

2-Allylphenol (2-AP) is a synthetic phenylpropanoid, structurally related to cardanol, thymol, and ortho-eugenol. Phenylpropanoids are described in the literature as being capable of promoting biological activity. Due to the similarity between 2-AP and other bioactive phenylpropanoids, the present research aims at evaluating the antioxidant, antinociceptive, and anti-inflammatory potential of 2-AP in silico, in vitro, and in vivo. At 30 min prior to the start of in vivo pharmacological testing, administration of 2-AP (25, 50, 75, and 100 mg/kg i.p.), morphine (6 mg/kg i.p.), dexamethasone (2 mg/kg s.c.), or vehicle alone was performed. In the acetic acid-induced abdominal writhing tests, pretreatment with 2-AP significantly reduced the number of abdominal writhes, as well as decreased licking times in the glutamate and formalin tests. Investigation of the mechanism of action using the formalin model led to the conclusion that the opioid system does not participate in its activity. However, the adenosinergic system is involved. In the peritonitis tests, 2-AP inhibited leukocyte migration and reduced releases of proinflammatory mediators TNF-α and IL-1ß. In vitro antioxidant assays demonstrated that 2-AP presents significant ability to sequester superoxide radicals. In silico docking studies confirmed interaction between 2-AP and the adenosine A2a receptor through hydrogen bonds with the critical asparagine 253 residues present in the active site. Investigation of 2-AP demonstrated its nociception inhibition and ability to reduce reactive oxygen species. Its interaction with A2a receptors may well be related to proinflammatory cytokines TNF-α and IL-1ß reduction activity, corroborating its antinociceptive effect.

Sigmoid gallstone ileus: a challenging diagnosis

Gallstone ileus is a rare (1%­4%) complication of gallstone disease. Gallstones entering the gastrointestinal tract by penetration may cause obstruction at any point along their course through the tract; however, they have a predilection to obstruct the smaller-caliber lumen of the small intestine (80.1%) or stomach (14.2%). The condition is seen more commonly in the elderly who often have significant co-morbidities. Gallstone ileus causing large bowel obstruction is rare. We report the case of a 95-year-old woman who presented with a history of abdominal pain without fever, nausea, vomiting, or diarrhea. Computed tomography of the abdomen and pelvis with oral contrast revealed a high-density structure within the lumen of the distal sigmoid colon, initially suspected to be a foreign body. Medical management failed and surgical intervention was not possible. Autopsy revealed peritonitis and a rupture of the sigmoid colon at the site of a cylindrical stone found impacted in an area of fibrotic narrowing with multiple diverticula. A necrotic, thick-walled gallbladder had an irregular stone in its lumen that was a fracture match with the stone in the sigmoid. Adhesions, but no discrete fistula, were identified between the gallbladder and the adjacent transverse colon. The immediate cause of death was peritonitis caused by colonic perforation by the gallstone impacted at an area of diverticular narrowing. To our knowledge, such autopsy findings have not been previously reported.

Pancreatitis aguda: fisiopatología y manejo inicial / Acute pancreatitis: pathophysiology and initial management

Resumen La pancreatitis aguda es un trastorno intracelular del calcio en las células pancreáticas, el cual constituye la vía final común de múltiples estímulos etiopatogénicos y puede desencadenar cambios necroinflamatorios locales, efectos multisistémicos y compromiso en órganos distantes. Todo esto lleva a los pacientes a múltiples complicaciones por disfunción orgánica e infección. El diagnóstico adecuado y oportuno, el abordaje según severidad y la optimización de la terapia nutricional, así como una adecuada analgésica, reanimación hídroelectrolítica, detección de disfunción orgánica y de complicaciones locales e infecciosas, determinan el desenlace clínico de dicha patología. Se realizó una revisión narrativa incluyendo estudios clínicos, guías de manejo, protocolos y revisiones pertinentes, y se aporta un enfoque desde el punto de vista de medicina crítica para el abordaje inicial de esta patología.

Abstract Acute pancreatitis is an intracellular calcium disorder in pancreatic cells, which constitutes the final common pathway of multiple etiopathogenic stimuli and can trigger local necroinflammatory changes, multisystemic effects and compromise distant organs. All of this leads to multiple complications due to organ dysfunction and infection in patients. The adequate and opportune diagnosis, the approach according to severity and the optimization of the nutritional therapy; as well as an adequate analgesic, hydroelectrolytic resuscitation, the detection of organic dysfunction and of local and infectious complications, determine the clinical outcome of this pathology. A narrative review was carried out including clinical studies, management guidelines, protocols and reviews. An initial approach for this pathology, from the critical medicine point of view, is provided.

Role of moxifloxacin-dexamethasone in cardiac histomorphometric findings among Wistar rats from infected mothers.

PURPOSE: To investigate cardiac changes in young rats, whose mothers underwent autogenic fecal peritonitis, during organogenesis phase and to evaluate the role of intravenous administration of moxifloxacin and dexamethasone in preventing infection-related cardiac changes. METHODS: A prospective histomorphometric study was performed on 29 hearts of Wistar four-month old rats. Animals were divided into three groups: Negative Control Group (NCG) included 9 subjects from healthy mothers; Positive Control Group (PCG) included 10 subjects from mothers with fecal peritonitis (intra-abdominal injection of 10% autogenic fecal suspension in the gestational period) and did not receive any treatment; and Intervention Group (IG), with 10 animals whose infected mothers received moxifloxacin and dexamethasone treatment 24 hours after induction of fecal peritonitis. RESULTS: Nuclear count was higher in the IG group as compared to PCG (p = 0.0016) and in NCG as compared to PCG (p = 0.0380). There was no significant difference in nuclear counts between NCG and IG. CONCLUSION: Induced autogenic fecal peritonitis in pregnant Wistar rats determined myocardial changes in young rats that could be avoided by the early administration of intravenous moxifloxacin and dexamethasone.

The Bisindole Alkaloid Caulerpin, from Seaweeds of the Genus Caulerpa, Attenuated Colon Damage in Murine Colitis Model.

Caulerpin (CLP), an alkaloid from algae of the genus Caulerpa, has shown anti-inflammatory activity. Therefore, this study aimed to analyze the effect of CLP in the murine model of peritonitis and ulcerative colitis. Firstly, the mice were submitted to peritonitis to evaluate which dose of CLP (40, 4, or 0.4 mg/kg) could decrease the inflammatory infiltration in the peritoneum. The most effective doses were 40 and 4 mg/kg. Then, C57BL/6 mice were submitted to colitis development with 3% dextran sulfate sodium (DSS) and treated with CLP at doses of 40 and 4 mg/kg. The disease development was analyzed through the disease activity index (DAI); furthermore, colonic tissue samples were submitted to histological analysis, NFκB determination, and in vitro culture for cytokines assay. Therefore, CLP at 4 mg/kg presented the best results, triggering improvement of DAI and attenuating the colon shortening and damage. This dose was able to reduce the TNF-α, IFN-γ, IL-6, IL-17, and NFκB p65 levels, and increased the levels of IL-10 in the colon tissue. Thus, CLP mice treatment at a dose of 4 mg/kg showed promising results in ameliorating the damage observed in the ulcerative colitis.

Role of moxifloxacin-dexamethasone in cardiac histomorphometric findings among Wistar rats from infected mothers

Abstract Purpose: To investigate cardiac changes in young rats, whose mothers underwent autogenic fecal peritonitis, during organogenesis phase and to evaluate the role of intravenous administration of moxifloxacin and dexamethasone in preventing infection-related cardiac changes. Methods: A prospective histomorphometric study was performed on 29 hearts of Wistar four-month old rats. Animals were divided into three groups: Negative Control Group (NCG) included 9 subjects from healthy mothers; Positive Control Group (PCG) included 10 subjects from mothers with fecal peritonitis (intra-abdominal injection of 10% autogenic fecal suspension in the gestational period) and did not receive any treatment; and Intervention Group (IG), with 10 animals whose infected mothers received moxifloxacin and dexamethasone treatment 24 hours after induction of fecal peritonitis. Results: Nuclear count was higher in the IG group as compared to PCG (p = 0.0016) and in NCG as compared to PCG (p = 0.0380). There was no significant difference in nuclear counts between NCG and IG. Conclusion: Induced autogenic fecal peritonitis in pregnant Wistar rats determined myocardial changes in young rats that could be avoided by the early administration of intravenous moxifloxacin and dexamethasone.

Photobiomodulation reduces neutrophil migration and oxidative stress in mice with carrageenan-induced peritonitis.

The purpose of the study is to investigate the effects of two doses of photobiomodulation (PBM) on inflammatory parameters including cell migration and oxidative stress in carrageenan-induced peritonitis models. Twenty-eight mice were divided into four groups: saline; untreated carrageenan (Cg; inflammation induced); and PMB treatment groups L1 and L5 (inflammation induced with carrageenan followed by laser irradiation at 1 and 5 J/cm2, respectively). After 30 min of inducing inflammation, laser irradiation was administered every hour, for 4 h. Peritoneal fluid was collected for analyses. The total leukocyte number in the peritoneal fluid in L1 (4.33 ± 2.34) and L5 (4.95 ± 2.86) after PBM was lower than that in Cg (10.93 ± 5.15 cells/ml). The average differential count of neutrophils in the Cg was 9.46 ± 4.31 cells/ml, which was higher than that in L1 (3.7 ± 2.08) and L5 (4.94 ± 2.57). Myeloperoxidase activity was also lower in L1 (1.89 ± 0.43) and L5 (4.84 ± 2.62) than in Cg (22.92 ± 4.52 UMPO/ml). Malondialdehyde content was lower in L1 (137.5 ± 12.33) and L5 (169.6 ± 22.77) than in Cg (345.7 ± 65.67 nmol/ml). Glutathione peroxidase concentration was significantly higher in L1 (155.2 ± 12.43) and L5 (145.9 ± 9.585) than in Cg (79.75 ± 9.567 µ/ml). Nitrite concentration was lower in L1 (0.3317 µM ± 0.0669) and L5 (0.2429 µM ± 0.0232) than in Cg (0.8380 µM ± 0.01615). Laser irradiation at 1 and 5 J/cm2 reversed the inflammation (as indicated by neutrophil infiltration and oxidative stress).

Structural design, synthesis and substituent effect of hydrazone-N-acylhydrazones reveal potent immunomodulatory agents.

4-(Nitrophenyl)hydrazone derivatives of N-acylhydrazone were synthesized and screened for suppress lymphocyte proliferation and nitrite inhibition in macrophages. Compared to an unsubstituted N-acylhydrazone, active compounds were identified within initial series when hydroxyl, chloride and nitro substituents were employed. Structure-activity relationship was further developed by varying the position of these substituents as well as attaching structurally-related substituents. Changing substituent position revealed a more promising compound series of anti-inflammatory agents. In contrast, an N-methyl group appended to the 4-(nitrophenyl)hydrazone moiety reduced activity. Anti-inflammatory activity of compounds is achieved by modulating IL-1ß secretion and prostaglandin E2 synthesis in macrophages and by inhibiting calcineurin phosphatase activity in lymphocytes. Compound SintMed65 was advanced into an acute model of peritonitis in mice, where it inhibited the neutrophil infiltration after being orally administered. In summary, we demonstrated in great details the structural requirements and the underlying mechanism for anti-inflammatory activity of a new family of hydrazone-N-acylhydrazone, which may represent a valuable medicinal chemistry direction for the anti-inflammatory drug development in general.

Lectin obtained from the red seaweed Bryothamnion triquetrum: Secondary structure and anti-inflammatory activity in mice.

Seaweeds are sources of biomolecules with biological activities and pharmacological potential - for example, lectins, a group of proteins that can bind reversibly to carbohydrates or compounds containing them. The aim of this study was to elucidate the structural properties of a lectin extracted from the red seaweed Bryothamnion triquetrum (BtL) and to investigate its anti-inflammatory activity in mice. The lectin was purified by precipitation with ammonium sulfate and ion-exchange chromatography. Its secondary structure and tryptophan (Trp) microenvironment were analyzed by circular dichroism spectroscopy and steady-state fluorescence spectroscopy, respectively. The anti-inflammatory effect was evaluated by means of paw edema induced by carrageenan or dextran, myeloperoxidase activity in paw tissue, and by measurement of leukocyte and neutrophil migration and cytokine quantification in a peritonitis model. The secondary structure of BtL is mostly composed of ß-strands and unordered conformation, and it is quite resistant to extremes of pH and temperature, preserving the exposure of Trp residues under these conditions. In an assessment of biological activities, groups of mice were subjected to pretreatment with BtL before the inflammatory stimulus. BtL had anti-inflammatory effects in the models tested, and hence may be considered a molecule with potential to be used in the pharmaceutical industry.

Selecting the right gate to identify relevant cells for your assay: a study of thioglycollate-elicited peritoneal exudate cells in mice.

OBJECTIVE: In this study, we investigate the diversity and modulation of leukocyte populations represented in the gates defined by size and granularity at different time points of thioglycollate-induced peritonitis in mouse. RESULTS: The inflammatory cells were distributed into four regions (R1-R4) of a data plot graph defined by cell size and granularity. R1 and R2 contained agranular cells that were small in size and predominately included T (CD3+) lymphocytes along with B (B220+) lymphocytes. Macrophages (F4/80+) were the predominant cells found in the R3 region. However, these cells were present in all regions, albeit at a lower frequency in R1 and R2. Granulocytes (Gr1+) were mainly distributed in R3 and R4. The wide distribution of F4/80+ and Gr1+ cells may reflect the recruitment and activation state of the different macrophage and granulocyte populations. Based on these observations, size and granularity may contribute to an initial step in the analysis and sorting of thioglycollate-elicited peritoneal exudate cells. However, the developmental stage and cell activation state may interfere with cell segregation using size and granularity as parameters.

Aspidosperma pyrifolium Has Anti-Inflammatory Properties: An Experimental Study in Mice with Peritonitis Induced by Tityus serrulatus Venom or Carrageenan.

Scorpions of the genus Tityus are responsible for the majority of envenomation in Brazil, the Tityus serrulatus species being the most common and dangerous in South America. In this approach, we have investigated the ability of the aqueous extract from the leaves of Aspidosperma pyrifolium in reducing carrageenan-induced inflammation and the inflammation induced by T. serrulatus envenomation in mice. We also evaluated the cytotoxic effects of this extract, using the 3-(4,5-dimethythiazol-2-yl)-2,5-diphenyl-2H-tetrazolium (MTT) assay and the results revealed that the extract is safe. Analysis by High Performance Liquid Chromatography coupled with Diode Array Detector (HPLC-DAD) and Liquid Chromatography Coupled with Mass Spectrometry with Diode Array Detection (LC-DAD-MS) showed one major chemical component, the flavonoid rutin and phenolics compounds. For in vivo studies in carrageenan-induced peritonitis model, mice received extracts, dexamethasone, rutin or saline, before administration of carrageenan. For venom-induced inflammation model, animals received T. serrulatus venom and were, simultaneously, treated with extracts, antivenom, rutin or saline. The extract and rutin showed a reduction in the cell migration into the peritoneal cavity, and in the same way the envenomated animals also showed reduction of edema, inflammatory cell infiltration and vasodilation in lungs. This is an original study revealing the potential action of A. pyrifolium against inflammation caused by Tityus serrulatus venom and carrageenan, revealing that this extract and its bioactive molecules, specifically rutin, may present potential anti-inflammatory application.

The anti-inflammatory and antinociceptive activity of albumins from Crotalaria retusa seeds.

Seeds of Crotalaria retusa L. are used in popular medicine because of their pharmacological properties. The albumin fraction obtained from its seeds contains lectin, a protein known to have analgesic and anti-inflammatory properties. Thus, albumins extracted from C. retusa were investigated for their anti-inflammatory and antinociceptive effects. The intraperitoneal administration of different doses of albumins (5, 10 or 20mg/kg) significantly inhibited the mice paw edema induced by carrageenan (maximum inhibition rate of 80.9% at four hours, 20mg/kg), and this event was followed by diminishing paw myeloperoxidase measurements. Albumins (20mg/kg) also inhibited neutrophil migration into the peritoneal cavity induced by carrageenan. However, no effect was observed in the dextran-induced paw edema and abdominal contortions induced by acetic acid. Moreover, albumins (20mg/kg) significantly reduced the second (inflammatory) phase of the licking time induced by formalin. The detection of heammaglutinating activity against human erythrocytes in albumins evidences the presence of lectin in seeds of C. retusa. Our data showed that seeds of C. retusa had anti-inflammatory and antinociceptive properties and such activities are probably due to the inhibitory effect on neutrophil migration of lectin present in albumins.

Morinda citrifolia lipid transfer protein 1 exhibits anti-inflammatory activity by modulation of pro- and anti-inflammatory cytokines.

Previous reports have demonstrated that a thermostable lipid transfer protein isolated from noni seeds (McLTP1; 9.4kDa) displays anti-nociceptive and anti-inflammatory activities. This work aimed to investigate the underlying mechanisms of the anti-inflammatory activity of McLTP1 in mice. The protein was solubilised in sterile saline (0.9% NaCl) immediately before the treatment of mice by oral or intraperitoneal routes at doses of 8mg/kg. Given orally or intraperitoneally, McLTP1 significantly inhibited (p<0.05) cell migration in experimental models of carrageenan-induced peritonitis and the formation of paw oedema induced by carrageenan and dextran. Additionally, McLTP1 demonstrated the ability to significantly inhibit the production of the cytokines IL-1ß, IL-6, and TNF-α (p<0.05) and to promote an increase in the production of the anti-inflammatory cytokine IL-10. The treatment of mice with McLTP1 by the oral or i.p route reduced pancreatic injury and activities of amylase, lipase, and pancreatitis-associated lung injury. This study suggested that the observed anti-inflammatory effects of McLTP1 can be related to modulation of pro- and anti-inflammatory cytokine levels.