Febrile neutropenia management in pediatri onco-hematologic patients: a systematic review / Manejo da neutropenia febril em pacientes pediátricos onco-hematológicos: uma revisão sistemática / Manejo de la neutropenia febril en pacientes pediátricos oncohematológicos: una revisión sistemática

Background and Objectives: cancer cases are gradually increasing, and most treatments still cause several adverse reactions, such as myelosuppression. When neutrophils decline, febrile neutropenia (FN) can be triggered, considered an oncological emergency, leaving patients susceptible to infections. Therefore, it is necessary to determine the best treatment, seeking to reduce the risk of complications. The purpose of this review is to identify, in literature, randomized clinical studies that compare different treatments for FN in pediatric onco-hematological patients. Content: a systematic search was carried out on the PubMed database, for randomized clinical studies, from 2009 to 2019, in English, using "Febrile Neutropenia", "Pediatric", and "Therapeutics" as descriptors. A total of 233 articles were found, seven of which were selected for review. The most described antimicrobial for FN treatment was Piperacillin/Tazobactam (PIP/TAZ) and its use is justified by its spectrum of action to cover the most frequent microorganisms in patients with FN. The possibility of using oral antimicrobials may be an alternative and should be analyzed. The description of the risk classification criteria is essential to guide the therapy, and new tools, such as the stewardship, add safety to patient care. Conclusion: the most used antimicrobial to treat FN was PIP/TAZ, and the establishment of standardized risk classification scores in pediatric onco-hematological patients is essential to guide clinical management in FN treatment.(AU)

Justificativa e objetivos: os casos de câncer estão aumentando gradativamente, e a maioria dos tratamentos ainda causa várias reações adversas, como a mielossupressão. Com o declínio dos neutrófilos, pode-se desencadear a neutropenia febril (NF), considerada uma emergência oncológica, deixando o paciente suscetível a infecções. Portanto, é necessário determinar o melhor tratamento, visando reduzir o risco de complicações. O objetivo desta revisão é identificar, na literatura, estudos clínicos randomizados que comparem diferentes tratamentos para NF em pacientes onco-hematológicos pediátricos. Conteúdo: foi realizada busca sistemática na base de dados PubMed, de estudos clínicos randomizados, no período de 2009 a 2019, na língua inglesa, utilizando como descritores "Febrile Neutropenia", "Pediatric" e "Therapeutics". Foram encontrados 233 artigos, dos quais sete foram selecionados para revisão. O antimicrobiano mais descrito para o tratamento com FN foi Piperacilina / Tazobactam (PIP / TAZ) e seu uso justifica-se por seu espectro de ação para cobrir os microrganismos mais frequentes em pacientes com FN. A possibilidade de uso de antimicrobianos orais pode ser uma alternativa e deve ser analisada. A descrição dos critérios de classificação de risco é essencial para orientar a terapia, e novas ferramentas, como o stewardship, agregam segurança ao atendimento ao paciente. Conclusão: o antimicrobiano mais utilizado para tratar FN foi o PIP / TAZ, e o estabelecimento de escores de classificação de risco padronizados em pacientes onco-hematológicos pediátricos é essencial para orientar o manejo clínico no tratamento de FN.(AU)

Antecedentes y objetivos: los casos de cáncer están aumentando gradualmente y la mayoría de los tratamientos aún causan varias reacciones adversas, como la mielosupresión. Cuando los neutrófilos disminuyen, se puede desencadenar la neutropenia febril (FN), considerada una emergencia oncológica, dejando a los pacientes susceptibles a infecciones. Por tanto, es necesario determinar el mejor tratamiento, buscando reducir el riesgo de complicaciones. El propósito de esta revisión es identificar, en la literatura, estudios clínicos aleatorizados que comparen diferentes tratamientos para la FN en pacientes pediátricos oncohematológicos. Contenido: se realizó una búsqueda sistemática en la base de datos PubMed, de estudios clínicos aleatorizados, de 2009 a 2019, en inglés, utilizando como descriptores "Febrile Neutropenia", "Pediatric" y "Therapeutics". Se encontraron un total de 233 artículos, siete de los cuales fueron seleccionados para revisión. El antimicrobiano más descrito para el tratamiento de FN fue Piperacilina / Tazobactam (PIP / TAZ) y su uso se justifica por su espectro de acción para cubrir los microorganismos más frecuentes en pacientes con FN. La posibilidad de utilizar antimicrobianos orales puede ser una alternativa y debe analizarse. La descripción de los criterios de clasificación de riesgo es fundamental para orientar la terapia, y nuevas herramientas, como la rectoría, añaden seguridad a la atención al paciente. Conclusión: el antimicrobiano más utilizado para tratar la FN fue la PIP / TAZ, y el establecimiento de puntuaciones estandarizadas de clasificación de riesgo en pacientes pediátricos oncohematológicos es fundamental para orientar el manejo clínico en el tratamiento de la FN.(AU)

Influencia adrenérgica y colinérgica sobre la producción de especies reactivas de oxígeno en neutrófilos humanos* / Adrenergic and cholinergic influence on the production of reactive oxygen species in human neutrophils*

RESUMEN Objetivos. Determinar y comparar el efecto de fármacos agonistas adrenérgicos y colinérgicos sobre la producción de especies reactivas de oxígeno (ROS) en neutrófilos de individuos sanos. Materiales y métodos. Se tomaron muestras de sangre total de cinco participantes para purificar los neutrófilos mediante el método de gelatina. Se midió la producción de ROS por quimioluminiscencia (QLM) usando un contador de centelleo y forbol-12-miristato-13-acetato (PMA) como estímulo. También se realizaron pruebas sin PMA para medir la producción espontánea. Posteriormente, con el mismo método se midió la formación de ROS en presencia de nicotina (agonista colinérgico), salbutamol y clonidina (agonistas adrenérgicos), cada uno en concentraciones de 10-2 M, 10-3 M, 10-4 M y 10-5 M. Se calculó el área integrada bajo las curvas de QLM y se halló el porcentaje de inhibición o de estimulación según sea el caso. Se comparó el efecto provocado por las drogas con sus controles correspondientes y se realizó el análisis estadístico. Resultados. Se obtuvo una disminución de la producción de ROS como efecto de las sustancias estudiadas con una diferencia significativa entre los controles y el efecto producido a 10-2 M, 10-3 M y 10-4 M. Este efecto aumentó de intensidad conforme la concentración de las drogas se incrementó. Los mayores porcentajes de inhibición se mostraron a 10-2 M y 10-3 M. Salbutamol presentó los máximos valores con todas las concentraciones con diferencia significativa entre su inhibición y la generada por las demás drogas. Conclusiones. Los estímulos adrenérgico y colinérgico tienen un efecto inhibitorio de la producción de ROS en neutrófilos de individuos sanos.

ABSTRACT Objectives. To determine and compare the effect of adrenergic and cholinergic agonist drugs on the production of reactive oxygen species (ROS) in neutrophils of healthy individuals. Materials and Methods. Whole blood samples were taken from five participants to purify neutrophils using the gelatin method. The production of chemiluminescent (QLM) ROS was measured using a scintillation counter and phorbol-12-myristat-13-acetate (PMA) as a stimulus. Non-PLA tests were also conducted to measure spontaneous production. Subsequently, with the same method, ROS formation was measured in the presence of nicotine (cholinergic agonist), salbutamol, and clonidine (adrenergic agonists), each in concentrations of 10-2 M, 10-3 M, 10-4 M, and 10-5 M. The area integrated under the QLM curves was calculated and the percentage of inhibition or stimulation was found as the case may be. The effect of the drugs was compared with their corresponding controls and statistical analysis was carried out. Results. A decrease in the production of ROS was obtained as an effect of the substances studied with a significant difference between the controls and the effect produced at 10-2 M, 10-3 M, and 10-4 M . This effect increased in intensity as drug concentration increased. The highest percentages of inhibition were shown at 10-2 M and 10-3 M. Salbutamol presented the maximum values with all the concentrations with a significant difference between its inhibition and that generated by the other drugs. Conclusions. Adrenergic and cholinergic stimuli have an inhibitory effect on the production of ROS in neutrophils of healthy individuals.

Gliclazide reduced oxidative stress, inflammation, and bone loss in an experimental periodontal disease model

Abstract Objective The aim of this study was to evaluate the effects of gliclazide on oxidative stress, inflammation, and bone loss in an experimental periodontal disease model. Material and Methods Male albino Wistar rats were divided into no ligature, ligature, and ligature with 1, 5, and 10 mg/kg gliclazide groups. Maxillae were fixed and scanned using micro-computed tomography to quantify linear and bone volume/tissue volume (BV/TV) and volumetric bone loss. Histopathological, immunohistochemical and immunofluorescence analyses were conducted to examine matrix metalloproteinase-2 (MMP-2), cyclooxygenase 2 (COX-2), cathepsin K, members of the receptor activator of the nuclear factor kappa-Β ligand (RANKL), receptor activator of nuclear factor kappa-Β (RANK), osteoprotegerin (OPG) pathway, macrophage migration inhibitory factor (MIF), superoxide dismutase-1 (SOD-1), glutathione peroxidase-1 (GPx-1), NFKB p 50 (Cytoplasm), NFKB p50 NLS (nuclear localization signal), PI3 kinase and AKT staining. Myeloperoxidase activity, malondialdehyde and glutathione levels, while interleukin-1 beta (IL-1β) and tumor necrosis factor-alpha (TNF-α) levels were evaluated by spectroscopic ultraviolet-visible analysis. A quantitative reverse transcription polymerase chain reaction was used to quantify the gene expression of the nuclear factor kappa B p50 subunit (NF-κB p50), phosphoinositide 3-kinase (PI3k), protein kinase B (AKT), and F4/80. Results Micro-computed tomography showed that the 1 mg/kg gliclazide treatment reduced linear bone loss compared to the ligature, 5 mg/kg gliclazide, and 10 mg/kg gliclazide treatments. All concentrations of gliclazide increased bone volume/tissue volume (BV/TV) compared to the ligature group. Treatment with 1 mg/kg gliclazide reduced myeloperoxidase activity, malondialdehyde, IL-1β, and TNF-α levels (p≤0.05), and resulted in weak staining for COX-2, cathepsin k, MMP-2, RANK, RANKL, SOD-1, GPx-1,MIF and PI3k. In addition, down-regulation of NF-κB p50, PI3k, AKT, and F4/80 were observed, and OPG staining was strong after the 1 mg/kg gliclazide treatment. Conclusions This treatment decreased neutrophil and macrophage migration, decreased the inflammatory response, and decreased bone loss in rats with ligature-induced periodontitis.

Agonistic activity of tamoxifen, a selective estrogen-receptor modulator (SERM), on arthritic ovariectomized mice

Arthritis is positively associated with the decline of sex hormones, especially estrogen. Tamoxifen (TMX) is a selective estrogen receptor modulator, possessing agonist or antagonistic activity in different tissues. Thus, the objective of this study was to investigate the effect of TMX on the zymosan-induced arthritis model. Female Swiss normal and ovariectomized (OVX) mice were divided into groups and treated for five days with TMX (0.3, 0.9 or 2.7 mg/kg) or 17-β-estradiol (E2, 50 µg/kg). On the fifth day, arthritis was induced and 4 h later, leukocyte migration into joint cavities was evaluated. The neutrophil migration in OVX animals, but not in normal mice, treated with TMX (all tested doses) was significantly decreased compared with mice that received the vehicle (P≤0.05). Similarly, this effect was also demonstrated in the E2-treated group. Therefore, the present study demonstrates that TMX presented agonist effects in inhibiting neutrophil migration and preventing arthritis progression in OVX mice.

Schinus terebinthifolius raddi (Aroeira) and Orbignya phalerata mart. (Babassu) effect in cecorrahphy healing in rats

ABSTRACT PURPOSE: To evaluate the effect of Schinus terebinthifolius Raddi (aroeira) and Orbignya phalerata Mart. (babassu) in the healing process of cecorrhaphy in rats. METHODS : Fifty four rats were used, distributed into three groups randomly: aroeira, babassu and control, which were divided into three subgroups (six animals) according to the time of the deaths (7, 14, 21 days). All underwent the same surgical procedure, cecotomy and cecorrhaphy. The animals in group aroeira and babassu received daily dose of 100 mg/kg of hydroalcoholic extract and 50 mg/kg of aquous extract respectively, by gavage. The control group received only saline solution. The parameters evaluated were: macroscopic changes, ,resistance test to air insufflations and histological changes. RESULTS : All animals showed good healing without infection. All groups presented adhesions between cecum and neighboring organs. The resistance test insufflating of atmospheric air showed progressive increase of pressure according to the days in the aroeira group, and decrease in babassu group, without significant difference. Microscopy showed significant difference in the polymorphonuclear, hyperemia, angiogenesis, fibroblast proliferation and collagen histological variables in the 14th day. CONCLUSION : Hydroalcoholic extract of aroeira and the aqueous extract of babassu favored the healing process in cecorrhaphy in rats.

Effects of budlein A on human neutrophils and lymphocytes

ABSTRACT Sesquiterpene lactones (SLs) are the active constituents of a variety of medicinal plants used in traditional medicine for the treatment of inflammatory diseases and other ailments. Objective In this study, we evaluated whether budlein A modulates the activation of innate and adaptive immune cells such as neutrophils and lymphocytes. Material and Methods Our research group has investigated several plant species and several compounds have been isolated, identified, and their medical potential evaluated. Budlein A is a SL isolated from the species Aldama buddlejiformis and A. robusta (Asteraceae) and shows anti-inflammatory and anti-nociceptive activities. Advances in understanding how plant-derived substances modulate the activation of innate and adaptive immune cells have led to the development of new therapies for human diseases. Results Budlein A inhibited MPO activity, IL-6, CXCL8, IL-10, and IL-12 production and induces neutrophil apoptosis. In contrast, budlein A inhibited lymphocyte proliferation and IL-2, IL-10, TGF-β, and IFN-γ production, but it did not lead to cell death. Conclusions Collectively, our results indicate that budlein A shows distinct immunomodulatory effects on immune cells.

Chemical and cytological analysis of cerebral spinal fluid after intrathecal injection of hypodense fluorescein / Análise química e citológica do líquor após injeção intratecal de solução hipodensa de fluoresceína

ABSTRACT INTRODUCTION: Intrathecal fluorescein has been effective for topographic diagnosis of rhinoliquorrhea. Nonetheless, there are no reports on the study of cerebral spinal fluid (CSF) after use of intrathecal fluorescein. OBJECTIVE: A prospective study attempting to evaluate CSF through chemical and cytological analysis, after injection of fluorescein. METHODS: Prospective analysis of 24 samples of CSF after intrathecal injection of fluorescein for topographic diagnosis of CSF fistulae, collected at the time of puncture and after 24 and 48 h, divided by cellularity: Group 1, up to five cells, and Group 2, with more than five cells. RESULTS: The yellow-greenish color of CSF remained after 48 h in 36%, evidencing permanence of fluorescein. No changes in protein and glucose levels were observed between 0-24 h and 0-48 h. In group 2, an increase in cell count was observed between 24 h and 48 h (p = 0.019). In both groups, there was an increase of neutrophils between 0 and 48 h (p = 0.048) and a decrease between 24 and 48 h (p = 0.05). CONCLUSION: Intrathecal fluorescein provoked discreet meningeal reactions, such as an increase of cells between 24 and 48 h and an increase of neutrophils at 24 h, with a subsequent decrease at 48 h with no correlation with symptomatology.

RESUMO Introdução: A fluoresceína intratecal tem sido efetiva no diagnóstico topográfico da rinoliquorréia. Entretanto, não há estudos no líquor após o uso de fluoresceína intratecal. Objetivo: Estudo prospectivo visando avaliar o líquor, através de análise química e citológica, após injeção de fluoresceína. Método: Análise prospectiva de 24 punções após injeção intratecal de fluoresceína para diagnóstico topográfico de fístula liquórica, coletado no momento da punção, 24 e 48 horas, divididos pela celularidade: grupo 1, com até 5 células e grupo 2 com mais de 5 células. Resultado: A coloração amarelo-esverdeada do líquor permaneceu após 48 horas em 36%, evidenciando permanência de fluoresceína. Observou-se ausência de mudanças no nível de proteína e glicose entre 0-24 horas e 0-48 horas. No grupo 2, um aumento na contagem celular foi observado entre 24 e 48 horas (p = 0,019). No dois grupos juntos, observou-se um aumento de neutrófilos entre 0 e 48 horas (p = 0,048) e uma diminuição entre 24 e 28 horas (p = 0,05). Conclusão: Fluoresceína intratecal provocou discretas reações meníngeas, como o aumento de células entre 24 e 48 horas e aumento dos dos neutrófilos em 24 horas com uma subsequente dimi nuição em 48 horas sem correlação com sintomas.

Effect of hypertonic saline treatment on the inflammatory response after hydrochloric acid-induced lung injury in pigs

OBJECTIVES: Hypertonic saline has been proposed to modulate the inflammatory cascade in certain experimental conditions, including pulmonary inflammation caused by inhaled gastric contents. The present study aimed to assess the potential anti-inflammatory effects of administering a single intravenous dose of 7.5% hypertonic saline in an experimental model of acute lung injury induced by hydrochloric acid. METHODS: Thirty-two pigs were anesthetized and randomly allocated into the following four groups: Sham, which received anesthesia and were observed; HS, which received intravenous 7.5% hypertonic saline solution (4 ml/kg); acute lung injury, which were subjected to acute lung injury with intratracheal hydrochloric acid; and acute lung injury + hypertonic saline, which were subjected to acute lung injury with hydrochloric acid and treated with hypertonic saline. Hemodynamic and ventilatory parameters were recorded over four hours. Subsequently, bronchoalveolar lavage samples were collected at the end of the observation period to measure cytokine levels using an oxidative burst analysis, and lung tissue was collected for a histological analysis. RESULTS: Hydrochloric acid instillation caused marked changes in respiratory mechanics as well as blood gas and lung parenchyma parameters. Despite the absence of a significant difference between the acute lung injury and acute lung injury + hypertonic saline groups, the acute lung injury animals presented higher neutrophil and tumor necrosis factor alpha (TNF-α), interleukin (IL)-6 and IL-8 levels in the bronchoalveolar lavage analysis. The histopathological analysis revealed pulmonary edema, congestion and alveolar collapse in both groups; however, the differences between groups were not significant. Despite the lower cytokine and neutrophil levels observed in the acute lung injury + hypertonic saline group, significant differences were not observed among the treated and non-treated groups. ...

The effect of visfatin on inflammatory reaction in uterus of LPS-induced rats / Efecto de visfatina en la reacción inflamatoria en el útero de ratas inducidas por LPS

The present study was to investigate the effects of visfatin on the morphological structure and function of the rat uterus during inflammation. The expression and distribution of visfatin, morphological structure, eosinophils (EOS), myeloperoxidase (MPO) and cytokines in the uterus of the LPS-induced rat were studied using hematoxylin-eosin staining (HE), immunohistochemical methods, western blots and enzyme-linked immunosorbent assay (ELISA). The present study showed that visfatin positive cells dispersed widely in the uterus, and strong positive staining was observed mainly in the cell cytoplasm. Compared with saline group, in visfatin group, more uterine glands were found, EOS increased, and the difference was significant (P<0.05), MPO reduced, and the difference was significant (P<0.01). In addition, visfatin was able to increase the secretion of IL-1b, IL-6, and TNF-a (P<0.01). Compared with LPS group, in vifatin+LPS group, the uterine glands of the lamina propria increased, the myometrium became thinner, the number of EOS and MPO reduced obviously, but the difference was not significant (P>0.05), and after LPS stimulated body, visfatin decrease the level of IL-1b, IL-6, TNF-a (P<0.01). The above results suggest that visfatin could affect the morphological structure of rat uterus; Visfatin could modulate the inflammatory response in rats' uterus by regulating the quantity of inflammatory cells, such as EOS and MPO, and the level of inflammatory cytokines, such as IL-1b, IL-6, TNF-a.

El objetivo del presente estudio fue investigar los efectos de la visfatina sobre la estructura morfológica y la función del útero de la rata durante la inflamación. Se estudiaron la expresión y distribución de la visfatina, la estructura morfológica, eosinófilos, mieloperoxidasa y citoquinas en el útero de rata mediante la tinción de H&E, métodos inmunohistoquímicos, Western blots y ELISA. El estudio mostró que las células visfatina positivas se dispersan ampliamente en el útero, junto a una fuerte tinción positiva, principalmente en el citoplasma de la célula. En comparación con el grupo control, en el grupo visfatina, se encontraron más glándulas uterinas, se observó un aumento de EOS y la diferencia fue significativa (p<0,05), MPO reducida siendo esta diferencia también significativa (p<0,01). Además, la visfatina fue capaz de aumentar la secreción de IL-1b, IL-6 y TNF-a (P<0,01). En comparación con el grupo LPS, visfatina+grupo LPS, las glándulas uterinas de la lámina propia aumentaron, se observó un miometrio más delgado, y número reducido de EOS y MPO, sin embargo, la diferencia no fue significativa (P>0,05). Después de estímulo LPS en el cuerpo, se registró un nivel menor de visfatina en IL-1b, IL-6, TNF-a (P<0,01). Los resultados anteriores sugieren que visfatina podría afectar a la estructura morfológica del útero de rata. Además, podría modular la respuesta inflamatoria en el útero mediante la regulación de la cantidad de células inflamatorias, tales como EOS y MPO.

Minimal endoscopic changes in non-erosive reflux disease / Alterações endoscópicas mínimas na doença do refluxo não erosiva

BACKGROUND: Gastroesophageal reflux disease is an increasingly common condition worldwide causing a considerable economic impact. More than half the patients with clinical symptoms of reflux disease display no mucosal erosions on esophagogastroduodenoscopy, making it impossible to confirm the diagnosis without further investigations. AIM: To evaluate the correlation between minimal endoscopic changes on white-light esophagogastroduodenoscopy (carditis, mucosal thickening and invisibility of vessels) and histologic changes observed in distal esophageal biopsies in a sample of patients with symptoms suggestive of reflux disease, and to verify the specificity of these symptoms for non-erosive reflux disease. METHODS: Retrospective, cross-sectional study based on information retrieved from a digital database at a Brazilian hospital for the period March-October, 2012. The sample consisted of previously untreated, non-smoking subjects aged >18 years with symptoms suggestive of reflux disease but no esophageal erosions, submitted to esophagogastroduodenoscopy and distal esophageal biopsy. RESULTS: The final sample included 23 subjects. The most frequently observed change was invisibility of vessels (n=21; 91.3%), followed by mucosal thickening (n=15; 65.2%) and carditis (n=5; 21.7%). The correlation coefficient between each variable and the anatomopathological diagnosis was 0.386 for body mass index, 0.479 for mucosal thickening, -0.116 for invisibility of vessels, 0.306 for carditis and 0.462 for hiatal hernia. CONCLUSION: All patients displayed minimal endoscopic changes on esophagogastroduodenoscopy, but only mucosal thickening revealed a moderately significant correlation with severity of esophagitis, although increased body mass index values and the presence of hiatal hernia were also associated. .

RACIONAL: Doença do refluxo gastroesofágico é condição cada vez mais comum em todo o mundo causando impacto econômico considerável. Mais da metade dos pacientes com sintomas clínicos da doença não apresentam erosões endoscópicas da mucosa, o que torna impossível confirmar o diagnóstico sem outras investigações. OBJETIVO: Avaliar a correlação entre mudanças mínimas endoscópicas em endoscopia digestiva alta de luz branca (cardite, espessamento da mucosa e invisibilidade de vasos) e as alterações histológicas observadas em biópsias distais de uma amostra de pacientes com sintomas sugestivos de doença do refluxo, e para verificar a especificidade desses sintomas para a doença não-erosiva. MÉTODOS: Estudo retrospectivo, transversal, com base em informações obtidas a partir de uma base de dados digital em um hospital brasileiro no período de março/outubro de 2012. A amostra foi composta por indivíduos não tratados previamente, não fumantes, >18 anos, com sintomas sugestivos de doença do refluxo, mas sem erosões esofágicas submetidos à endoscopia digestiva alta e biópsia de esôfago distal. RESULTADOS: A amostra final incluiu 23 indivíduos. A alteração mais frequente foi invisibilidade dos vasos (n=21; 91,3%), seguido por espessamento de mucosa (n=15; 65,2%) e cardite (n=5; 21,7%). O coeficiente de correlação entre cada variável e o diagnóstico anatomopatológico foi 0,386 para o índice de massa corporal, 0,479 para espessamento de mucosa, -0,116 para a invisibilidade de vasos, 0,306 para carditis e 0,462 para hérnia hiatal. CONCLUSÃO: Todos os pacientes apresentaram alterações endoscópicas mínimas, mas apenas espessamento da mucosa revelou correlação moderadamente significativa com a gravidade da esofagite, apesar do aumento dos valores no índice de massa corporal e da presença de hérnia hiatal também estarem associados. .

Oral administration of curcumin (Curcuma longa) can attenuate the neutrophil inflammatory response in zymosan-induced arthritis in rats

PURPOSE: To evaluate the effect of curcumin in the acute phase of zymosan-induced arthritis. METHODS: Twenty-eight male rats were subjected to intra-articular infiltration of zymosan of both knees and, in four the infiltration was made with saline. The animals were divided into five groups second received every six hours by gavage: corn oil by (positive and negative control); curcumin (100 mg/kg); prednisone 1 mg/kg/day; prednisone 8 mg/kg. All animals were sacrificed after six, 12, 24 and 48 hours of the infiltration. The knees were removed for evaluation of neutrophil infiltration. The number of neutrophils was counted by computer-assisted analysis of the images. The neutrophil infiltrate was stratified into four grades: 0 = normal; + = mild; ++/+++ = moderate; > ++++ = severe. The results were compared using the Mann-Whitney test and the variance by Kruskal-Wallis test adopting a significance level of 5% (p<0.05). RESULTS: Curcumin reduces inflammatory activity in the first six hours after zymosan-induced arthritis when compared to saline (p<0.01). This was also observed in animals subjected to administration of prednisone (1 mg/kg) and those treated with prednisone (8 mg/kg). Curcumin was more effective than lower doses of prednisone in the first six hours after induction of the arthritis. After 12, 24 and 48 hours, curcumin does not have the same anti-inflammatory effects when compared to prednisone. After 48 hours, prednisone is more effective than curcumin in reducing the inflammatory infiltrate regardless of the dose of prednisone used. CONCLUSION: Oral administration of curcumin reduces inflammation in the first six hours after experimentally zymosan-induced arthritis. .

Comparative immunomodulation potential of Tinospora cordifolia (Willd.) Miers ex Hook. F., Tinospora sinensis (Lour.) Merrill and Tinospora cordifolia growing on Azadirachta indica A. Juss.

Guduchi has been widely used in the traditional medicine as an immunomodulator. Description of guduchi in Ayurvedic literature resemble with T. sinensis rather than with commonly available T. cordifolia and hence this may be used as substitutes for T. sinensis. T. cordifolia growing on Azadirachta indica commonly called Neem-guduchi has more immunomodulatory potential. Thus, immunomodulatory activity of three Tinospora spp. was assessed by checking humoral and cell mediated immune responses to the antigenic challenges with sheep RBCs and by neutrophil adhesion tests on albino Wistar rats using Guduchi-Satwa, a well known dosage form. Results revealed that Neem-guduchi possesses higher immunomodulatory potential at the dose of 300 mg/kg, po and validated the traditional claim. Hence, Neem-Guduchi can be employed in immunomodulatory formulation prepared using guduchi.

Diclofenac topical gel in excisional wounds maintain heal quality and reduce phlogistic signals1

PURPOSE: To investigate diclofenac topical gel as an alternative to reduce phlogistic signals and maintain quality of wound repair. METHODS: Fifteen Wistar rats were used in this study; four excisional wounds were performed on the dorsum of each animal. Once in a day, cranial wounds received topical diclofenac gel administration and caudal wounds were washed with isotonic saline. After seven, 14 and 21 postoperative days, five animals were randomly chosen for macroscopic and microscopic wound analysis. RESULTS: On the 7th day: diclofenac wounds showed significant higher scab formation, however showed less phlogistic signal; diclofenac wounds had larger area and had less neutrophil invasion. On the 14th day: No area difference was noted and diclofenac wounds showed less hyperemia and phlogistic signals; diclofenac wounds showed greater keratinocytes invasion. On the 21st day: Almost all wounds were closed and there were no difference regarding the type of scar formation; diclofenac wounds showed greater monocytes invasion and lower angiogenesis level. No difference was noted in any postoperative day regarding fibroblast invasion, collagen deposit quantity and quality. CONCLUSION: Diclofenac topical gel is capable of reducing phlogistic signals and do not cause fibroblast or keratinocyte downregulation thus do not lead to excisional wound healing impairment. .

Tissue response evaluation of the mucosa of the tympanic cavity of guinea pigs, when receiving biodegradable implant

PURPOSE: To evaluate the tissue response of the mucosa of the tympanic cavity of guinea pigs, when receiving biodegradable implant. METHODS: A total of 20 male guinea pigs were divided into 2 groups. After paracentesis in both ears, a biodegradable polymer of poly lactic-co-glycolic acid was implanted in only one middle ear. Histological analysis using neutrophil exudate and vascular neoformation (acute inflammation) and fibroblast proliferation and mononuclear inflammatory cells (chronic inflammation) as parameters was performed after 10 and 30 days of survival (groups 1 and 2, respectively). RESULTS: Four ears in group 1 and 7 in group 2 had an increase of neutrophil exudate. Vascular neoformation occurred in ears with or without the implant, in both groups. Fibroblast proliferation and mononuclear inflammatory cells (lymphocytes and macrophages) increased in ears with implant in group 2. CONCLUSION: The tissue response by histological analysis of the mucosa of the tympanic cavity of guinea pigs, when receiving biodegradable implant, showed no statistically significant difference between ears with or without the implant. .

Oxidative response of neutrophils to platelet-activating factor is altered during acute ruminal acidosis induced by oligofructose in heifers

Reactive oxygen species (ROS) production is one of the main mechanisms used to kill microbes during innate immune response. D-lactic acid, which is augmented during acute ruminal acidosis, reduces platelet activating factor (PAF)-induced ROS production and L-selectin shedding in bovine neutrophils in vitro. This study was conducted to investigate whether acute ruminal acidosis induced by acute oligofructose overload in heifers interferes with ROS production and L-selectin shedding in blood neutrophils. Blood neutrophils and plasma were obtained by jugular venipuncture, while ruminal samples were collected using rumenocentesis. Lactic acid from plasma and ruminal samples was measured by HPLC. PAF-induced ROS production and L-selectin shedding were measured in vitro in bovine neutrophils by a luminol chemiluminescence assay and flow cytometry, respectively. A significant increase in ruminal and plasma lactic acid was recorded in these animals. Specifically, a decrease in PAF-induced ROS production was observed 8 h after oligofructose overload, and this was sustained until 48 h post oligofructose overload. A reduction in PAF-induced L-selectin shedding was observed at 16 h and 32 h post oligofructose overload. Overall, the results indicated that neutrophil PAF responses were altered in heifers with ruminal acidosis, suggesting a potential dysfunction of the innate immune response.

Inhibition of Janus activated kinase-3 protects against myocardial ischemia and reperfusion injury in mice

Recent studies have documented that Janus-activated kinase (JAK)-signal transducer and activator of transcription (STAT) pathway can modulate the apoptotic program in a myocardial ischemia/reperfusion (I/R) model. To date, however, limited studies have examined the role of JAK3 on myocardial I/R injury. Here, we investigated the potential effects of pharmacological JAK3 inhibition with JANEX-1 in a myocardial I/R model. Mice were subjected to 45 min of ischemia followed by varying periods of reperfusion. JANEX-1 was injected 1 h before ischemia by intraperitoneal injection. Treatment with JANEX-1 significantly decreased plasma creatine kinase and lactate dehydrogenase activities, reduced infarct size, reversed I/R-induced functional deterioration of the myocardium and reduced myocardial apoptosis. Histological analysis revealed an increase in neutrophil and macrophage infiltration within the infarcted area, which was markedly reduced by JANEX-1 treatment. In parallel, in in vitro studies where neutrophils and macrophages were treated with JANEX-1 or isolated from JAK3 knockout mice, there was an impairment in the migration potential toward interleukin-8 (IL-8) and monocyte chemoattractant protein-1 (MCP-1), respectively. Of note, however, JANEX-1 did not affect the expression of IL-8 and MCP-1 in the myocardium. The pharmacological inhibition of JAK3 might represent an effective approach to reduce inflammation-mediated apoptotic damage initiated by myocardial I/R injury.

Histological evaluation of bone response to pediatric endodontic pastes: an experimental study in guinea pig

This study aimed to evaluate by the intra-osseous implant technique the most commonly used materials for pulp therapy in pediatric dentistry: calcium hydroxide (CH), Guedes Pinto paste and CTZ paste, according to FDI (1980) and ANSI/ADA (1982) recommendations. Thirty guinea pigs, 10 for each material, divided into experimental periods of 4 and 12 weeks received one implant on each side of the lower jaw symphysis. The external lateral tube wall served as control for the technique. At the end of the observation periods, the animals were euthanized and specimens were prepared for routine histological examination. It was observed that CH and CTZ paste induced severe inflammation, a large amount of necrotic tissue, lymphocytes, foreign body cells and bone resorption, while Guedes Pinto Paste induced little or no inflammation in the 4-week observation period. After 12 weeks, the reactions to CH and Guedes Pinto paste were also absent/mild, presenting a general pattern of replacement by recently formed bone tissue while a moderate to severe inflammatory response was observed with CTZ paste. Guedes Pinto paste presented acceptable biocompatibility levels in both analyzed periods; CH only showed acceptable biocompatibility in the 12-week period while CTZ paste showed no biocompatibility in both periods. Among the tested materials, only Guedes Pinto paste presented an acceptable biocompatibility.

A pesquisa teve como objetivo avaliar a biocompatibilidade através da técnica de implantes intra-ósseos dos materiais utilizados em odontopediatria para tratamento pulpar: hidróxido de cálcio, pastas Guedes Pinto e CTZ, de acordo com as recomendações da FDI (1980) e ANSI/ADA(1982). Trinta guinea pigs, dez para cada material, divididos em períodos experimentais de 4 e 12 semanas receberam um implante em cada lado da sínfise mandibular. A parede lateral externa do copo serviu como controle para a técnica. No final dos períodos experimentais, os animais foram sacrificados e os espécimes preparados para o exame histológico de rotina. Observou-se que o hidróxido de cálcio e a pasta CTZ mostraram reação inflamatória severa, grande quantidade de tecido necrosado, linfócitos, células de corpo estranho e reabsorção óssea; enquanto a pasta Guedes Pinto induziu pouca ou nenhuma inflamação no período de 4 semanas. Após 12 semanas as reações para o hidróxido de cálcio e pasta Guedes Pinto foram ausentes/suaves apresentando um padrão geral de substituição por tecido ósseo neoformado, enquanto uma resposta inflamatória de moderada a severa foi observada para a pasta CTZ. A pasta Guedes Pinto apresentou níveis aceitáveis de biocompatibilidade nos dois períodos analisados; hidróxido de cálcio apresentou biocompatibilidade aceitável somente no período de 12 semanas e a pasta CTZ não mostrou biocompatibilidade em ambos os períodos. Entre estes, apenas a pasta Guedes Pinto apresentou níveis de biocompatibilidade nos dois períodos analisados.

Neutrophil predominance in induced sputum from asthmatic patients: Therapeutic implications and role of clara cell 16-KD protein / Predominio de neutrófilos en esputo inducido de asmáticos: Implicancias terapéuticas y rol de la proteína 16-kD de la célula clara

Eosinophil is considered to be a main protagonist in asthma; however, often discordances between clinical manifestations and response to treatment are observed. We aimed to determine the occurrence of neutrophil predominance in asthma and to identify its characteristics on the basis of clinical-functional features, induced sputum cellular pattern and soluble molecules, to guide the appropriated anti-inflammatory therapy. A total of 41 patients were included in randomized groups: 21-40 year-old, with stable mild-to-severe asthma, steroid-naïve and non-smokers. An induced sputum sample was obtained under basal conditions, a second one after treatment with budesonide (400 µg b.i.d.) or montelukast (10 mg/d) for six weeks, and a final one after a 4-week washout period. By cytospin we evaluated eosinophil (EP) or neutrophil predominance (NP), and in supernatant we determined LTE4, and CC16. Peak expiratory flow variability (PEFV) was measured. A total of 23/41 patients corresponded to EP and 18/41 patients to NP. The PEFV was higher in EP than in NP. LTE4 was higher with NP than with EP. No difference was found for CC16. Montelukast reduced the predominant cell in both subsets, whereas budesonide only reduced eosinophils in EP. Budesonide and montelukast reduced PEFV in EP but not in NP. Considering the total treated-samples in each subset, CC16 level increased significantly in EP. In conclusion: a NP subset of asthmatic patients was identified. These patients show a lower bronchial lability; the leukotriene pathway is involved which responds to anti-leukotriene treatment. This phenotype shows a poor recovery of CC16 level after treatment.

El eosinófilo es considerado la célula protagonista principal en el asma; sin embargo, a menudo se observan discordancias entre las manifestaciones clínicas y la respuesta de los pacientes al tratamiento. Nos propusimos determinar la ocurrencia de predominio de neutrófilos en el asma e identificar las características clínico-funcionales, el patrón celular y las moléculas solubles del esputo inducido, para guiar el tratamiento apropiado anti-inflamatorio. Se incluyeron 41 pacientes: 21 a 40 años de edad, con asma estable leve a grave, no tratados con esteroides tópicos ni sistémicos y no fumadores. Se obtuvo una muestra de esputo inducido en condiciones basales, una segunda muestra después del tratamiento al azar con budesonida (400 µg dos veces al día) o el montelukast (10 mg/d) durante seis semanas, y una final después de un período de lavado de 4 semanas. En el frotis por citocentrifugado se evaluó el predominio de eosinófilos (EP) o neutrófilos (NP), y en el sobrenadante se determinó LTE4, y CC16. Se midió la variabilidad del flujo espiratorio máximo (PEFV). Un total de 23/41 pacientes correspondieron al EP y 18/41 pacientes con NP. El PEFV fue mayor en el EP que en NP. LTE4 fue mayor en NP que en EP. No se encontraron diferencias de los niveles de CC16 en ambos grupos. Montelukast redujo la célula predominante en ambos subgrupos, mientras que budesonida sólo redujo los eosinófilos en EP. Tanto budesonida como montelukast redujeron PEFV en EP, pero no en NP. El nivel de CC16 aumentó significativamente en el EP luego del tratamiento antiinflamatorio. En conclusión: se identificó un subgrupo de asmáticos NP que presentan una menor labilidad bronquial, la vía de los leucotrienos parece estar involucrada y responde al tratamiento anti-leucotrienos. Este fenotipo muestra una escasa recuperación del nivel de CC16 posterior al tratamiento.

The effect of montelukast in a model of gouty arthritis induced by sodium monourate crystals / Efecto de montelukast en un modelo de artritis gotosa inducido por cristales de monourato de sodio

Non-steroidal anti-inflammatory drugs (NSAIDS) are the first line of therapy in acute gouty arthritis. NSAIDs inhibit the cyclooxygenase pathway, but not the lipooxygenase activity and can have many adverse effects and thus have a limited effect on the control of inflammation in this disease. In this work we studied the effect of montelukast on the cellular inflammatory infiltrate in a model of murine arthritis induced by sodium monourate crystals (SMU), using a subcutaneous air cavity (air pouch) in BALB/c mice. Seven groups of BALB/c mice (n = 4) were distributed into five experimental groups and two inflammatory control groups, a positive and a negative one. Previous to SMU exposure, the experimental groups received montelukast (1 and 0.01 mg/Kg/w) and/or indomethacine (2.5 mg/Kg/w), followed by administration of SMU in the air pouch. The total and differential counts of inflammatory cells were analyzed after 2, 6, 12 and 24 hours. Montelukast, significantly reduced the total number of cells (p<0.05), with a predominant impact on polymorphonuclear over mononuclear cells, especially after 12 hours of the medication. The montelukast/indometacine combination showed an additive effect. Our data show that montelukast has an anti-inflammatory effect in the model of gouty arthritis. Consequently, anti-leukotrienes could represent a new and effective therapy, either isolated or combined with conventional therapy of gouty arthritis.

En artritis gotosa aguda las drogas antiinflamatorias no esteroideas son la primera línea terapéutica. Este tratamiento no es satisfactorio porque inhibe la ciclooxigenasa sin modificar la actividad de la lipooxigenasa, y puede acompañarse de numerosos efectos adversos. Investigamos el efecto de montelukast sobre el infiltrado celular inflamatorio en un modelo de artritis múrida inducida por cristales de monourato de sodio (MUS) en el modelo experimental de la bolsa de aire (air pouch). Siete grupos de ratones BALB/c (n = 4) fueron distribuidos en cinco grupos experimentales y dos grupos controles inflamatorios: positivo y negativo. Los grupos experimentales recibieron, montelukast (1 y 0,01 mg/Kg/p) y/o indometacina (2,5 mg/Kg/p) por vía oral, previo a la administración de MUS en la bolsa del aire. El conteo absoluto y diferencial de las células inflamatorias fue analizado después de 2, 6, 12 y 24 horas de tratamiento. El tratamiento con montelukast redujo significativamente el número total de células presentes en el infiltrado inflamatorio (p < 0,05), con un efecto mayor sobre polimorfonucleares que sobre las células mononucleares, y con un máximo efecto a las 12 horas después de la administración del medicamento. La combinación montelukast/indometacina mostró un efecto aditivo. Los resultados demuestran que montelukast tiene un efecto antiinflamatorio en el modelo de la artritis gotosa. Por lo tanto, los anti-leucotrienos podrían representar una nueva y eficaz terapia, aislada o en combinación con la terapéutica convencional, para la artritis gotosa.

Pulp capping materials exert an effect on the secretion of IL-1β and IL-8 by migrating human neutrophils

Pulp repair is a complex process whose mechanisms are not yet fully understood. The first immune cells to reach the damaged pulp are neutrophils that play an important role in releasing cytokines and in phagocytosis. The objective of this study was to analyze the effect of different pulp-capping materials on the secretion of interleukin-1 beta (IL-1β) and interleukin-8 (IL-8) by migrating human neutrophils. Neutrophils were obtained from the blood of three healthy donors. The experimental groups were calcium hydroxide [Ca(OH)2], an adhesive system (Single Bond), and mineral trioxide aggregate (MTA). Untreated cells were used as control. Transwell chambers were used in performing the assays to mimic an in vivo situation of neutrophil chemotaxis. The pulp-capping materials were placed in the lower chamber and the human neutrophils, in the upper chamber. The cells were counted and the culture medium was assayed using ELISA kits for detecting and quantifying IL-1β and IL8. The data were compared by ANOVA followed by Tukey's test (p < 0.05). The secretion of IL-8 was significantly higher in all groups in comparison to the control group (p < 0.05). The adhesive system group showed higher IL-8 than the MTA group (p < 0.05). The secretion of IL-1β was significantly greater only in the MTA group (p < 0.001). It was concluded that only MTA is able to improve the secretion of IL-1β, and all materials tested increased IL-8 secretion. These results combined with all the other biological advantages of MTA indicate that it could be considered the material of choice for dental pulp capping.