Antitumor and immunomodulatory activity of fucoidan from the brown alga Lessonia trabeculata (Lessoniaceae) on breast cancer spheroids / Actividad antitumoral e inmunomoduladora de fucoidan del alga parda Lessonia trabeculata (Lessoniaceae) en esferoides de cáncer de mama

Abstract Introduction: The therapeutic benefits of the brown algae fucoidan in the treatment of breast cancer have attracted considerable interest in recent years. However, research using spheroids which provide relevant results in trials for antitumor and immunomodulatory products because they adequately simulate the tumor microenvironment, is limited. Objective: To evaluate the antitumor and immunomodulatory activity of Lessonia trabeculata fucoidan (LtF), native to the Peruvian Sea, on two types of multicellular tumor spheroids. Methods: The study was conducted from January to December 2021. Two types of spheroides were elaborated: from 4T1 tumor cells (MTS), and from 4T1 tumor cells+mouse splenocytes (MTSs). The antitumor activity of LtF was evaluated in MTS by quantifying cell viability with MTT. Immunomodulatory activity was determined in MTSs using the IC50 for two types of treatment: simple, fucoidan alone (LtF) and combined, fucoidan+doxorubicin (LtF+Dox). Pro-inflammatory (TNF-α, IL-6) and anti-inflammatory (IL-10, TGF-β) cytokine production was quantified by sandwich ELISA 72 h after treatment. Dox was used as positive control in all assays. Results: LtF exerted antitumor activity as evidenced by increased necrotic zone and cell debris formation compared to the untreated control. Antitumor activity was concentration dependent between 100 and 6 000 μg/ml. In MTSs, simple treatment increased IL-6 and decreased IL-10 and TGF-β production. The combined treatment significantly reduced TGF-β production. In both treatments and Dox, there was an increase in IL-6 compared to the untreated control. The highest production of IL-10 and TGF-β was observed in the untreated control, compatible with a highly immunosuppressive tumor microenvironment. Conclusions: LtF is a good candidate for the treatment of breast cancer and can immunomodulate the tumor microenvironment alone or in combination with Dox.

Resumen Introduccción: Los beneficios terapéuticos del fucoidan de algas pardas en el tratamiento del cáncer de mama han despertado gran interés en los últimos años. Sin embargo, las investigaciones con esferoides son limitadas, éstos proporcionan resultados relevantes en ensayos de productos antitumorales e inmunomoduladores porque simulan adecuadamente el microambiente tumoral. Objetivo: Evaluar la actividad antitumoral e inmunomoduladora del fucoidan de Lessonia trabeculata (LtF), nativa del Mar Peruano, en dos tipos de esferoides tumorales multicelulares. Métodos: El estudio se realizó de enero a diciembre de 2021. Se elaboraron dos tipos de esferoides: con células tumorales 4T1 (MTS) y con células tumorales 4T1+esplenocitos de ratón (MTSs). La actividad antitumoral de LtF se evaluó en MTS cuantificando la viabilidad celular con MTT. La inmunomodulación se determinó en MTSs utilizando la IC50 para dos tipos de tratamiento: simple, fucoidan solo (LtF) y combinado, fucoidan+doxorubicina (LtF+Dox). La producción de citoquinas proinflamatorias (TNF-α, IL-6) y antiinflamatorias (IL-10, TGF-β) se cuantificó mediante ELISA sándwich 72 h post-tratamiento. En todos los ensayos se utilizó Dox como control positivo. Resultados: En los MTS, el LtF ejerció actividad antitumoral evidenciada por aumento de la zona necrótica y formación de restos celulares respecto al control no tratado. La actividad antitumoral fue concentración-dependiente entre 100 y 6 000 μg/ml. En los MTSs, con el tratamiento simple se incrementó IL-6 y disminuyeron IL-10 y TGF-β. El tratamiento combinado redujo significativamente la producción de TGF-β. Los dos tratamientos y Dox incrementaron IL-6 respecto al control no tratado. La mayor producción de IL-10 y TGF-β se observó en los no tratados, compatible con un microambiente tumoral altamente inmunosupresor. Conclusiones: El LtF es un buen candidato para tratar el cáncer de mama y puede inmunomodular el microambiente tumoral solo o en combinación con Dox.

NLRP3 inflammasome mechanism and therapeutic targets in diabetic cardiomyopathy.

Diabetic cardiomyopathy (DCM) is a serious complication of diabetes caused by oxidative stress, inflammation, insulin resistance, myocardial fibrosis, and lipotoxicity; its nature is insidious, complex and difficult to treat. NLRP3 inflammasome triggers the maturation and release of pro-inflammatory cytokines, participates in pathophysiological processes such as insulin resistance and myocardial fibrosis, in addition to being closely related to the development and progression of diabetic cardiomyopathy. The development of inhibitors targeting specific aspects of inflammation suggests that NLRP3 inflammasome can be used to treat diabetic cardiomyopathy. This paper aims to summarize NLRP3 inflammasome mechanism and therapeutic targets in diabetic cardiomyopathy, and to provide new suggestions for the treatment of this disease.

La cardiomiopatía diabética es una complicación grave de la diabetes causada por estrés oxidativo, inflamación, resistencia a la insulina, fibrosis miocárdica y lipotoxicidad. Se trata de un padecimiento insidioso, complejo y difícil de tratar. El inflamasoma NLRP3 desencadena la maduración y liberación de citoquinas proinflamatorias, participa en procesos fisiopatológicos como la resistencia a la insulina y la fibrosis miocárdica, además de estar estrechamente relacionado con la aparición y progresión de la cardiomiopatía diabética. El desarrollo de inhibidores dirigidos a aspectos específicos de la inflamación sugiere que el inflamasoma NLRP3 puede utilizarse para tratar la cardiomiopatía diabética. Este artículo pretende resumir el mecanismo y las dianas terapéuticas del inflamasoma NLRP3 en la cardiomiopatía diabética, así como aportar nuevas sugerencias para el tratamiento de esta enfermedad.

Mecanismos y dianas terapéuticas del inflamasoma NLRP3 en la miocardiopatía diabética / NLRP3 inflammasome mechanism and therapeutic targets in diabetic cardiomyopathy

Resumen La cardiomiopatía diabética es una complicación grave de la diabetes causada por estrés oxidativo, inflamación, resistencia a la insulina, fibrosis miocárdica y lipotoxicidad. Se trata de un padecimiento insidioso, complejo y difícil de tratar. El inflamasoma NLRP3 desencadena la maduración y liberación de citoquinas proinflamatorias, participa en procesos fisiopatológicos como la resistencia a la insulina y la fibrosis miocárdica, además de estar estrechamente relacionado con la aparición y progresión de la cardiomiopatía diabética. El desarrollo de inhibidores dirigidos a aspectos específicos de la inflamación sugiere que el inflamasoma NLRP3 puede utilizarse para tratar la cardiomiopatía diabética. Este artículo pretende resumir el mecanismo y las dianas terapéuticas del inflamasoma NLRP3 en la cardiomiopatía diabética, así como aportar nuevas sugerencias para el tratamiento de esta enfermedad.

Abstract Diabetic cardiomyopathy (DCM) is a serious complication of diabetes caused by oxidative stress, inflammation, insulin resistance, myocardial fibrosis, and lipotoxicity; its nature is insidious, complex and difficult to treat. NLRP3 inflammasome triggers the maturation and release of pro-inflammatory cytokines, participates in pathophysiological processes such as insulin resistance and myocardial fibrosis, in addition to being closely related to the development and progression of diabetic cardiomyopathy. The development of inhibitors targeting specific aspects of inflammation suggests that NLRP3 inflammasome can be used to treat diabetic cardiomyopathy. This paper aims to summarize NLRP3 inflammasome mechanism and therapeutic targets in diabetic cardiomyopathy, and to provide new suggestions for the treatment of this disease.

Estrogen deficiency influences TNF-α and IL-1ß gene expression in the odontogenic region of dental hypofunctional condition / A deficiência de estrógeno influencia a expressão gênica de TNF-α e IL-1ß na região odontogênica de dentes em hipofunção

Objetivo: Evidências científicas sugerem que a deficiência de estrógeno e fatores genéticos influenciam o desenvolvimento do sistema estomatognático. Este estudo teve como objetivo avaliar a influência da deficiência de estrógeno na expressão gênica de TNF-α, IL-1ß, IL-6 e IL-10 durante o desenvolvimento dentário em modelo murino. Material e Métodos: Ratas Wistar Hannover foram divididas em dois grupos de acordo com a intervenção recebida: Grupo Hipoestrogenismo - cirurgia de ovariectomia e Grupo Controle - cirurgia fictícia. Para avaliar o desenvolvimento dentário, o incisivo inferior foi escolhido. O modelo de hipofunção dos incisivos inferiores foi realizado por ajuste incisal. O incisivo homólogo exercia hiperfunção dentária. Os animais foram avaliados durante todo o período puberal. Após a eutanásia, as hemimandíbulas foram removidas para avaliar a expressão gênica do TNF-α, IL-1ß, IL-6 e IL-10 na região odontogênica dos incisivos por meio de PCR em tempo real. Foi realizado o teste de Kruskal-Wallis e o pós-teste de Dunn. O nível de significância foi de 5%. Resultados: Houve diferenças estatisticamente significativas na expressão gênica de TNF-α e IL-1ß entre os grupos hipoestrogenismo e controle sob condição de hipofunção dentária (p=0,0084, p=0,0072, respectivamente). Conclusão: A deficiência de estrógeno influencia a expressão gênica de TNF-α e IL-1ß na região odontogênica de dentes hipofuncionais (AU)

Objective: Scientific evidence suggests that estrogen deficiency and genetic factors have an influence on the development of the stomatognathic system. This study aimed to evaluate the influence of estrogen deficiency on the gene expression of TNF-α, IL-1ß, IL-6 and IL-10 during dental development in a murine model. Material and Methods: Wistar Hannover rats were divided into two groups according to the intervention received: Hypoestrogenism Group - ovariectomy surgery and Control Group - fictitious surgery. To evaluate the dental development, the lower incisor was chosen. The mandibular incisor hypofunction model was performed by incisal adjustment. The homologous incisor exerted a hyperfunction. The animals were evaluated throughout the pubertal period. After euthanasia, the hemimandibles were removed to evaluate the gene expression of the TNF-α, IL-1ß, IL-6 and IL-10 in the odontogenic region of the incisors through real time PCR. Kruskal-Wallis test and Dunn's posttest were performed. The level of significance was 5%. Results: There were statistically significant differences of TNF-α and IL-1ß gene expression between the hypoestrogenism and control groups under hypofunction condition (p=0.0084, p=0.0072, respectively). Conclusion: Estrogen deficiency influences TNF-α and IL-1ß gene expression in the odontogenic region of the hypofunctional teeth. (AU)

Efecto del extracto hidroalcohólico de la planta Lampaya medicinalis Phil . sobre marcadores inflamación inducidos por ácidos grasos en hepatocitos humanos HepG2

RESUMEN Introducción: la enfermedad de hígado graso no alcohólico (EHGNA) se caracteriza por la acumulación de gotas lipídicas (GL) y sobre expresión de la proteína de GL Perilipina 1 (PLIN1) en los hepatocitos. En su patogénesis y progresión participan NF-ĸB, caspasa-1 y citoquinas proinflamatorias como IL-1β. La medicina popular del norte de Chile utiliza la planta Lampaya medicinalis Phil. (Verbenaceae) contra enfermedades. Objetivo: evaluar el efecto de un extracto hidroalcohólico de lampaya (EHL) sobre la expresión de marcadores inflamatorios y proteínas asociadas a las GL en hepatocitos tratados con ácidos grasos. Métodos: se incubó hepatocitos HepG2 con 0,66 mM de ácido oleico (AO) y 0,33 mM de ácido palmítico (AP) por 24 o 48 horas en presencia o no de EHL. Se evaluó la expresión proteica de NF-ĸB, PLIN1 y caspasa-1 por Western blot y la expresión de ARNm de IL-1β por qPCR. Resultados: los hepatocitos tratados por 48 h con AO/AP mostraron un aumento en la expresión de IL-1β que fue revertido por la co-incubación con EHL. Conclusión: estos antecedentes aportan nueva evidencia respecto a la actividad biológica del EHL en un modelo de alteraciones metabólicas e inflamatorias, asociadas a la EHGNA, inducidas por AO/AP en hepatocitos humanos.

ABSTRACT Introduction: Nonalcoholic fatty liver disease (NAFLD) is characterized by the accumulation of lipid droplets (LD) and overexpression of the LD-associated protein Perilipin 1 (PLIN1). NF-ĸB, caspase-1 and proinflammatory cytokine such as IL-1β participate in the pathogenesis and progression of NAFLD. Traditional medicine in northern Chile uses the plant Lampaya medicinalis Phil. (Verbenaceae) against diseases. Objective: To evaluate the effect of a hydroalcoholic extract of lampaya (HEL) on the expression of inflammatory markers and LD-associated proteins in hepatocytes treated with fatty acids. Methods: HepG2 hepatocytes were incubated with 0.66 mM oleic acid (OA) and 0.33 mM palmitic acid (PA) for 24 or 48 h in the presence or not of HEL. The protein expression of NF-ĸB, PLIN1 and caspase-1 was evaluated by Western blot while the mRNA expression of IL-1β was assessed by qPCR. Results: hepatocytes treated for 48 h with OA/AP showed an increase in IL-1β expression that was reversed by co-incubation with HEL. Conclusion: These antecedents provide new evidence regarding the biological activity of HEL in a model of metabolic and inflammatory alterations, associated with NAFLD, induced by OA/PA in human hepatocytes.

Polissacarídeos de Lentinula edodes suprimem a expressão de citocinas pró-inflamatórias e de colite em camundongos / Lentinula edodes polysaccharides suppressed pro-inflammatory cytokines expression and colitis in mice

ABSTRACT Background: Polysaccharides from edible mushrooms possess immunomodulatory, anti-inflammatory, and anti-tumor activities. Recent studies indicated that necroptosis plays a role in the pathogenesis of inflammatory diseases and mediates increased expression of inflammatory cytokines. Objective: Therefore, it is imperative to determine the impact of polysaccharide extract from Lentinula edodes (L. edodes) on inflammatory cytokines in experimental model of colitis in mice. Methods: Female C57BL/6 mice divided into three or four mice per group were used for this study. Polysaccharide sample was orally administered to mice prior to (7 days) and during colitis induction with 2.5% dextran sodium sulfate (7 days), followed by additional 3 days of administration. Changes in body weight and colon length were used as markers for colitis, and pro-inflammatory cytokines and tumor necrosis factor receptor 1 (TNFR1) expressions, as well as necroptosis were analyzed in the colon of colitis mice. Data obtained were analysed by Tukey-Kramer and two-tailed standard t tests. Results: The results indicated that the polysaccharide sample suppressed colitis in mice using effects on the body weight and colon length as markers. Also, it was demonstrated that necrostatin-1, a specific inhibitor of necroptosis, suppressed the expression of interleukin (IL)-8, a pro-inflammatory chemokine, in Caco-2 cells induced necroptosis induced by zVAD and TNF-α, an indication that necroptosis may be involved in the expression of pro-inflammatory cytokines. Moreover, the polysaccharide sample suppressed the expression of pro-inflammatory cytokines such as tumor necrosis factor (TNF)-α, IL-6, IL-1β, and interferon (IFN)-γ in the colon of mice. Conclusion: These results suggested that the suppressive effects of the polysaccharide sample on inflammatory cytokines expression may contribute to its anti-colitis effect, and so may serve as a potent therapeutic agent against inflammatory bowel disease.

RESUMO Contexto: Polissacarídeos de cogumelos comestíveis possuem atividades imunomodulatórias, anti-inflamatórias e anti-tumorais. Estudos recentes indicaram que a necroptose desempenha um papel na patogênese de doenças inflamatórias e regula o aumento da expressão de citocinas inflamatórias. Objetivo: Torna-se imprescindível determinar o impacto do extrato de polissacarídeo de Lentinula edodes (L. edodes) em citocinas inflamatórias em modelo experimental de colite em camundongos. Métodos: Foram utilizados para este estudo os camundongos C57BL/6 femininos divididos em três ou quatro camundongos por grupo. A amostra de polissacarídeo foi administrada oralmente em camundongos antes (7 dias) e durante a indução de colite com sulfato de dextran sulfato de sódio (7 dias), seguido por 3 dias adicionais de administração. Alterações no peso corporal e comprimento do cólon foram utilizadas como marcadores para colite, e citocinas pró-inflamatórias e tumores receptor fator 1 (TNFR1), bem como necroptose foram analisadas no cólon de camundongos colite. Os dados obtidos foram analisados por testes Tukey-Kramer e testes padrão t de duas caudas. Resultados: Os resultados indicaram que a amostra de polissacarídeo suprimiu colite em camundongos usando efeitos sobre o peso corporal e o comprimento do cólon como marcadores. Além disso, foi demonstrado que a necrostatina-1, inibidora específica da necroptose, suprimiu a expres são de interleucina (IL)-8, uma quimiocina pró-inflamatória, em células caco-2 induzidas necroptose induzidas por zVAD e TNF-α, uma indicação de que a necroptose pode estar envolvida na expressão de citocinas pro-inflamatórias. Além disso, a amostra de polissacarídeo suprimiu a expressão de citocinas pró-inflamatórias, como o fator de necrose tumoral (TNF)-α, IL-6, IL-1β e interferon (IFN)-γ no cólon dos camundongos. Conclusão: Esses resultados sugeriram que os efeitos supressivos da amostra de polissacarídeo na expressão de citocinas inflamatórias podem contribuir para o seu efeito anti-colite, podendo, portanto, servir como um potente agente terapêutico contra doença inflamatória intestinal.

Inflamación en la hernia del disco intervertebral / Inflammation in the intervertebral disc herniation

Hasta no hace muchos años, se consideró que el dolor ciático, en la hernia del disco intervertebral lumbar, estaba originado por la compresión sobre la raíz nerviosa. Sin embargo, la hernia del disco intervertebral lumbar presenta cuadros muy heterogéneos inexplicables con el simple compromiso mecánico. En los últimos años, numerosos estudios de inmuhistoquímica y de biología molecular han demostrado que el tejido herniado no es un material inerte sino, por el contrario, biológicamente muy activo con capacidad de expresar una serie de mediadores de inflamación entre los que destacan citoquinas proinflamatorias como la interleuquina1, interleuquina 6, interleuquina 8 y el factor de necrosis tumoral. La inflamación parece estar no solo inducida por la irritación química de las sustancias bioactivas liberadas por el núcleo pulposo, sino también mediante una respuesta autoinmune contra el mismo. Por tanto, además del factor mecánico, la mediación bioquímica tiene un papel importante en la fisiopatología del dolor ciático y de la radiculopatía. A través de una extensa revisión sistemática de la literatura se han investigado los mediadores celulares y moleculares que intervienen en dicho proceso inflamatorio alrededor de la hernia del disco intervertebral lumbar y su implicación en el dolor ciático (AU)

Up until fairly recently, it was thought that sciatic pain in the lumbar herniated disc was caused by compression on the nerve root. However, the lumbar herniated disc shows mixed pictures which are difficult to explain by simple mechanical compromise. In recent years various immunology, immunohistochemistry and molecular biology studies have shown that the herniated tissue is not an inert material, but rather it Is biologically very active with the capability of expressing a series of inflammatory mediators: cytokines such as interleukin-1, interleukin-6, interleuquin-8 and tumor necrosis factor being the ones which stand out. The inflammation is not only induced by the chemical irritation of the bioactive substances released by the nucleus pulposus but also by an autoimmune response against itself. Thus, in addition to the mechanical factor, the biomechanical mediation plays an important role in the pathophysiology of sciatic pain and of radiculopathy. Through a review of a wide range of literature, we researched the cellular molecular mediators involved in this inflammatory process around the lumbar herniated disc and its involvement in sciatic pain (AU)

Inflammation in the intervertebral disc herniation. / Inflamación en la hernia del disco intervertebral.

Up until fairly recently, it was thought that sciatic pain in the lumbar herniated disc was caused by compression on the nerve root. However, the lumbar herniated disc shows mixed pictures which are difficult to explain by simple mechanical compromise. In recent years various immunology, immunohistochemistry and molecular biology studies have shown that the herniated tissue is not an inert material, but rather it Is biologically very active with the capability of expressing a series of inflammatory mediators: cytokines such as interleukin-1, interleukin-6, interleuquin-8 and tumor necrosis factor being the ones which stand out. The inflammation is not only induced by the chemical irritation of the bioactive substances released by the nucleus pulposus but also by an autoimmune response against itself. Thus, in addition to the mechanical factor, the biomechanical mediation plays an important role in the pathophysiology of sciatic pain and of radiculopathy. Through a review of a wide range of literature, we researched the cellular molecular mediators involved in this inflammatory process around the lumbar herniated disc and its involvement in sciatic pain.

Influência do LED Violeta associado ou não ao gel clareador na inflamação, mineralização e maturação colágena do tecido dentinário e pulpar / Influence of violet LED associated or not with peroxide gel on inflammation, mineralization, and collagen fiber maturation in dentin and pulp tissue

Objetivo: Avaliar a influência do LED Violeta, associado ou não ao gel clareador a base de peróxido de hidrogênio (PH) a 17,5% no complexo dentino-pulpar de ratos. Materiais e métodos: Molares superiores de 80 ratos foram distribuídos nos grupos (n = 10): CONT ­ sem tratamento, PH ­ 1 aplicação de 30 minutos de PH a 17,5%, LED ­ 1 aplicação de 20 minutos do LED Violeta, e PH+LED - aplicação do PH e LED Violeta. Imediatamente (T0), e aos 7 (T1), 15 (T2) e 30 dias (T3) após o tratamento, os ratos foram eutanasiados e as maxilas processadas para avaliação histológica, imunoistoquímica (IL-17, IL-23 e osteocalcina), e de picrosírius red, sendo realizados os testes de Wilcoxon e Mann-Whitney e Teste-T pareado e Teste-T, respectivamente. (α = 0,05). Resultados: Necrose e infiltrado inflamatório severo foram observados nos grupos PH e PH+LED. Apenas o grupo PH+LED manteve a imunomarcação severa para IL-17 e IL-23, diferindo do grupo LED e PH que apresentaram moderada imunomarcação em T0. Os grupos PH e PH+LED apresentaram severa imunomarcação de OCN em T2 e moderada imunomarcação em T3. O grupo LED apresentou menor quantidade de fibras imaturas em T2 e T3 que o grupo CONT. Conclusão: A terapia com LED violeta não induziu inflamação e fibrose no tecido pulpar, apesar de acelerar a maturação das fibras de colágeno da dentina e, quando associada ao peróxido de hidrogênio, pode tornar os dentes mais sensíveis(AU)

Objective: Was to evaluated the influence of the Violet LED, associated or not with a 17.5% hydrogen peroxide (HP) bleaching gel in the dentin-pulp complex of rats. Materials and methods: Upper molars of eighty Wistar rats were distributed in the groups (n = 10): CONT - without treatment, HP - 1 application of 30 minutes of 17.5% HP, LED - 1 application of 20 minutes of the Violet LED, and HP+LED - application of HP and LED Violet. Immediately (T0), and at 7 (T1), 15 (T2) and 30 days (T3) after treatment, the rats were euthanized and the jaws were processed for histological, immunohistochemical evaluation (IL-17, IL-23 and osteocalcin), and picrosirius red, with Wilcoxon and Mann-Whitney tests and paired T-test and T-test, respectively (α = 0.05). Results: Necrosis and severe inflammatory infiltrate were observed in the PH and PH+LED groups. Only the PH+LED group maintained severe immunostaining for IL-17 and IL-23, differing from the LED and PH group which presented moderate T0 immunostaining. The PH and PH+LED groups presented severe immunostaining of OCN in T2 and moderate immunostaining in T3. The LED group had a lower amount of immature fibers in T2 and T3 than the CONT group. Conclusion: Violet LED therapy induced no inflammation and fibrosis in the pulp tissue, however accelerating the maturation of dentin collagen fibers and, when associated with hydrogen peroxide, can make the teeth more sensitive(AU)

Effect of coinfection with influenza virus and bacteria on host damage.

BACKGROUND: Influenza virus infection is often complicated by a bacterial infection, with this coinfection causing severe pneumonia. If not timely treated, the disease can cause death. OBJECTIVE: To demonstrate, in animal models, that coinfection with influenza virus and bacteria that affect the respiratory tract causes multisystemic damage. METHOD: Six groups of mice were formed: a control group, one infected with the influenza virus, two infected with bacteria: Haemophilus influenzae and Streptococcus pneumoniae, respectively; and two co-infected with influenza virus and Haemophilus influenzae or Streptococcus pneumoniae, respectively. RESULTS: Of the six groups of mice, only the group co-infected with influenza virus and Streptococcus pneumoniae showed damage to thoracic and abdominal organs. A decrease in serum cytokine levels was found in all study groups, which was more pronounced in the co-infected mice. CONCLUSIONS: The groups of mice infected with Streptococcus pneumoniae or influenza virus alone showed no damage, which indicates that coexistence of these infections caused the damage in the group of co-infected mice.

ANTECEDENTES: La infección por el virus de la influenza con frecuencia se complica con una infección bacteriana, coinfección que provoca cuadros graves de neumonía, la cual puede ocasionar la muerte si no es tratada en forma oportuna. OBJETIVO: Demostrar en modelos animales que la coinfección por el virus de la influenza y bacterias que afectan el tracto respiratorio ocasiona daño multisistémico. MÉTODO: Se formaron seis grupos de ratones: un grupo control, uno infectado de virus de la influenza, dos infectados de bacterias: Haemophilus influenzae y Streptococcus pneumoniae, respectivamente; y dos coinfectados de virus de la influenza y Haemophilus influenzae y Streptococcus pneumoniae, respectivamente. RESULTADOS: De los seis grupos de ratones, solo en el grupo coinfectado de virus de la influenza y Streptococcus pneumoniae se observó daño en órganos torácicos y abdominales. En todos los grupos se encontró disminución de los niveles séricos de las citocinas, mayor en los ratones coinfectados. CONCLUSIONES: Los grupos de ratones infectados solo de Streptococcus pneumoniae o el virus de la influenza no presentaron daños, lo cual indica que la coexistencia de estas infecciones fue la que ocasionó el daño en el grupo de ratones coinfectados.

Effect of coinfection with influenza virus and bacteria on host damage / Efecto de la coinfección por virus de la influenza y bacterias en el daño al hospedero

Abstract Background: Influenza virus infection is often complicated by a bacterial infection, with this coinfection causing severe pneumonia. If not timely treated, the disease can cause death. Objective: To demonstrate, in animal models, that coinfection with influenza virus and bacteria that affect the respiratory tract causes multisystemic damage. Method: Six groups of mice were formed: a control group, one infected with the influenza virus, two infected with bacteria: Haemophilus influenzae and Streptococcus pneumoniae, respectively; and two co-infected with influenza virus and Haemophilus influenzae or Streptococcus pneumoniae, respectively. Results: Of the six groups of mice, only the group co-infected with influenza virus and Streptococcus pneumoniae showed damage to thoracic and abdominal organs. A decrease in serum cytokine levels was found in all study groups, which was more pronounced in the co-infected mice. Conclusions: The groups of mice infected with Streptococcus pneumoniae or influenza virus alone showed no damage, which indicates that coexistence of these infections caused the damage in the group of co-infected mice.

Resumen Antecedentes: La infección por el virus de la influenza con frecuencia se complica con una infección bacteriana, coinfección que provoca cuadros graves de neumonía, la cual puede ocasionar la muerte si no es tratada en forma oportuna. Objetivo: Demostrar en modelos animales que la coinfección por el virus de la influenza y bacterias que afectan el tracto respiratorio ocasiona daño multisistémico. Método: Se formaron seis grupos de ratones: un grupo control, uno infectado de virus de la influenza, dos infectados de bacterias: Haemophilus influenzae y Streptococcus pneumoniae, respectivamente; y dos coinfectados de virus de la influenza y Haemophilus influenzae y Streptococcus pneumoniae, respectivamente. Resultados: De los seis grupos de ratones, solo en el grupo coinfectado de virus de la influenza y Streptococcus pneumoniae se observó daño en órganos torácicos y abdominales. En todos los grupos se encontró disminución de los niveles séricos de las citocinas, mayor en los ratones coinfectados. Conclusiones: Los grupos de ratones infectados solo de Streptococcus pneumoniae o el virus de la influenza no presentaron daños, lo cual indica que la coexistencia de estas infecciones fue la que ocasionó el daño en el grupo de ratones coinfectados.

Incremento de la viabilidad, producción de especies reactivas de oxígeno, IL-1 y TNF-α en células mononucleares de sangre periférica humana tratadas con fucoidan de Lessonia trabeculata

Las algas pardas constituyen una fuente de alto contenido de polisacáridos como los fucoidanos que poseen importantes propiedades inmunomoduladoras. El objetivo fue determinar la viabilidad de células mononucleares de sangre periférica humana (CMSPh), producción de óxido nítrico (NO), especies reactivas de oxígeno (ROS) y de las citoquinas proinflamatorias IL-1α, IL-6, TNF-α e IFN-γ en cultivos tratados con fucoidan de Lessonia trabeculata. Se empleó fucoidan de Lessonia trabeculata proveniente de la bahía San Nicolás de Marcona-Ica. Las CMSPh se aislaron empleando Ficoll-Hypaque, se distribuyeron a una concentración de 1x105 células/pocillo en medio RPMI-1640 completo y se trataron con diferentes concentraciones de fucoidan durante 24 y 48 h. La actividad citotóxica se determinó por la reducción de MTT, la producción de NO por la reacción de Griess y las ROS por la reducción del NBT. La producción de citoquinas se cuantificó por ELISA. El fucoidan de L. trabeculata estimuló la proliferación de CMSPh y produjo el incremento de ROS a concentraciones de 100-2000 μg/mL respecto al control (p<0.001), la reacción para nitritos resultó negativa. El fucoidan incrementó la producción de IL-1α y TNF-α a concentraciones de 100 y 10 μg/mL respectivamente, mientras que la producción de IL-6 e IFN-γ no mostró diferencias significativas. Se concluye que el fucoidan de L. trabeculata estimula la proliferación de CMSPh, producción de especies reactivas de oxígeno y las citoquinas proinflamatorias IL-1α y TNF-α que poseen importantes propiedades inmunomoduladoras.

Brown algae are a source of high content of polysaccharides such as fucoidans that have important immunomodulatory properties. The aim was to determine the viability of human peripheral blood mononuclear cells (hPBMC), production of nitric oxide (NO), reactive oxygen species (ROS) and the proinflammatory cytokines IL-1α, IL-6, TNF-α and IFN -γ in cultures treated with fucoidan from Lessonia trabeculata. Fucoidan from Lessonia trabeculata from San Nicolás de Marcona-Ica Bay was used. The hPBMC were isolated using Ficoll-Hypaque, distributed at a concentration of 1x105 cells/well in complete RPMI-1640 medium and treated with different concentrations of fucoidan for 24 and 48 h. The cytotoxic activity was determined by the reduction of MTT, NO production by the Griess reaction and ROS by the reduction of NBT. The production of cytokines was quantified by ELISA. The fucoidan of L. trabeculata stimulated the proliferation of hPBMC and produced the increase of ROS at concentrations of 100-2000 μg/mL with respect to the control (p <0.001), the reaction for nitrites was negative. Fucoidan increased the production of IL-1α and TNF-α at concentrations of 100 and 10 μg/mL respectively, while the production of IL-6 and IFN-γ did not show significant differences. It is concluded that the fucoidan of L. trabeculata stimulates the proliferation of hPBMC, production of reactive oxygen species and the proinflammatory cytokines IL-1α and TNF-α that possess important immunomodulatory properties.

[Circulating levels of MCP-1, VEGF-A, sICAM-1, sVCAM-1, sE-selectin and sVE-cadherin: Relationship with components of metabolic syndrome in young population]. / Concentraciones circulantes de MCP-1, VEGF-A, sICAM-1, sVCAM-1, sE-selectina y sVE-cadherina: su relación con componentes del síndrome metabólico en población joven.

INTRODUCTION: Inflammation and endothelial dysfunction are considered the primary manifestations of the cardiovascular disease. Studies have established a relationship among components of metabolic syndrome (MetS) with inflammatory markers and the loss of permeability, vasoconstriction and vasodilatation endothelial. OBJECTIVE: To determine the relationship among the concentrations of soluble endothelial dysfunction molecules and inflammation cytokines and components of the metabolic syndrome in young population. MATERIAL AND METHODS: A study was performed in 240 young adult students ages 18-28 years. To define the presence of clinical and metabolic alterations and MetS the modified ATP-III criteria was considered. In all subjects were determined sociodemographic characteristics, anthropometric measures and the metabolic profile. Circulating levels of MCP-1, VEGF-A, sICAM-1, sVCAM-1, sE-selectin and sVE-cadherin were determined by ELISA immunoassay (Bioscience). Statistical analysis was performed using STATA statistical software v. 9.2. RESULTS: From all the participants, 44.6% had obesity, 59.9% had abdominal obesity, 49.6% low HDL-c and 16.7% high levels triglycerids. The 16.25% of the population showed 3 or more components of the MetS. Elevated MCP-1, sICAM-1 and sE-selectin levels were linked to the presence of obesity. In a model adjusted by age-gender, high soluble levels of MCP-1 and VEGF-A were linked with abdominal obesity (OR=1.83; 1.02-3.28 and OR=2.03; 1.15-3.56, respectively), as well as to the presence of the 2 components of MetS. sVCAM-1 levels were associated with impaired glucose (OR=4.74; 1.32-17.0); sE-selectin with low HDL-c (OR=1.99; 1.05-3.75), although sICAM-1 and sVE-cadherin were associated with impaired systolic blood pressure (OR=4.04; 1.24-13.1 and OR=6.28; 1.90-20.7, respectively). CONCLUSION: Levels of circulating MCP-1 and VEGF-A were associated with adiposity, levels of sVCAM-1 with the presence of impaired glucose, sE-selectin with low HDL-c, while the levels of sICAM-1 and sVE-cadherin were associated with impaired systolic blood pressure in young adults independently of other traditional risk factors.

Peripheral blood leptin and resistin levels as clinical activity biomarkers in Mexican Rheumatoid Arthritis patients.

OBJECTIVE: To evaluate the association between the clinical activity of RA patients and serum adipocytokines (Leptin, Adiponectin and Resistin) and inflammatory cytokines. METHODS: All RA patients fulfilled ACR 1987 criteria and were treated with DMARDs. Adipocytokine and inflammatory cytokine levels were evaluated using ELISA. RESULTS: 121 patients were included in the study. Stratifying according to DAS28 (low, moderate and high activity), there were significant differences for Leptin, Resistin, IL-6 and IL-17, however, no differences were seen for Adiponectin, TNFα or IL-1ß. Clinical activity positively correlated with Leptin, Resistin, IL-17 and IL-6 levels, but not with Adiponectin, TNFα or IL-1ß. Adiponectin levels negatively correlated with TNFα and positively correlated with IL-1ß. IL-1ß positively correlated with IL-6 and negatively correlated with TNFα and IL-17. CONCLUSION: Circulating Leptin, Resistin, IL-6 and IL-17 levels positively correlate with RA clinical activity in a manner independent of the subject's BMI. Complex relationships between inflammatory cytokines were observed in RA patients suggesting that other metabolic or inflammatory factors could be involved.

Respuesta humoral IgM e IgG frente a antigenos de excreción-secreción de Schistosoma mansoni en murinos infectados y su relación con las citoquinas Il-10 y TNF-α / Humoral IgM and IgG against excretory-secretory antigens of infected murine with Schistosoma mansoni and its relationship with the cytokines Il-10 and TNF- α

El diagnóstico de fase en la esquistosomiasis es complejo. Se evaluó la respuesta IgM e IgGmurina contra productos de excreción-secreción de Schistosoma mansoni machos (PESGM), hembras (PESGH) y huevos (PESH), su relación con la histopatología y expresión de IL-10 y TNF-α, mediante ELISA, en suero de ratones Balb/c con 8 y 20 semanas de infección (RI8 y RI20) y Ratones sanos(RS). En RI8, se observaron granulomas constituidos por plasmocitos, macrófagos y neutrófilos, depósitos de colágeno alrededor de los granulomas y en la zona interna del huevo. En R20SI, se observaron fibroblastos alrededor del huevo y acúmulos de macrófagos y plasmocito, aumento de los depósitos de colágeno en áreas del granuloma. IgM sérica RI8, presentó un mayor porcentaje de positividad frente PESGH (35%), mientras que IgG el mayor porcentaje de positividad fue PESGH (60%) y PESH (30%). En R20SI, IgM fue 20% positiva frente PESH e IgG 10% frente PESGM, y 25% positiva con PESGH. No se observaron diferencias en IL-10 entre los RS y RI8. TNF-α en RS vs RI8 y RS vs RI20 fue diferente y estadísticamente significativo. Los PESGH podrían detectar fase aguda y PESH fase crónica. El empleo de varios antígenos sería de utilidad en el diagnóstico de fase.

The diagnosis of schistosomiasis phase is complex. The murine IgM and IgG response against excretory-secretory products of Schistosoma mansoni males (PESGM), females (PESGH) and eggs (PESH), its relation to histopathology and expression of IL-10 and TNF-α was assessed by ELISA in serum of Balb / c mice with 8 and 20 weeks of infection (RI8 and RI20) and healthy mice, RS. In RI8, were observed granulomas consisting of plasma cells, macrophages and neutrophils, deposits of collagen around the granulomas and internal area of the egg. In R20SI, fibroblasts around the egg and accumulation of macrophages and plasmocito, increased collagen deposits in areas of granuloma were observed.RI8 serum IgM had a higher percentage of positivity PESGH (35%), while the highest percentage of IgG positivity was PESGH (60%) and PESH (30%). In R20SI, was 20% IgM positive and IgG against PESH PESGM 10% against and 25% positive with PESGH. No differences in IL-10 between the RS and RI8 were observed. TNF-α in RS vs RI8 and RS vs RI20 was different and statistically significant. The PESGH could detect phase acute while PESH chronic phase. The use of several antigens would be useful in the diagnosis phase.

La teoría de la neuroinmunomodulación en enfermedades neurodegenerativas: nuevas evidencias científicas / Neuroinmmunomodulation in neurodegenerative diseases: new scientific evidence

Microglial cells play a major role in the innate immunity of the central nervous system. Alterations in the normal cross-talks between microglia and brain neuronal cells may lead to serious disturbances and neurodegenerative diseases. We have postulated that neuroinflammatory processes are a critical factor triggering the pathological cascade leading to neuronal degeneration. In our neuroimmunomodulation theory, external or internal damage signals activate microglial cells, producing cytotoxic factors that induce neuronal degeneration. These factors activate protein-kinases, that lead to tau hyperphosphorylation, and its consequent oligomerization. The tau aggregates released into the extracellular medium favor a positive feedback mechanism that determines neurodegeneration. Nowadays, natural components with a string anti-inflammatory activity and that cross the blood brain barrier appears as candidates for prevention and treatment of degenerative brain disorders such as Alzheimers'disease.

Las células microgliales juegan un papel importante en la inmunidad innata del sistema nervioso central. Las alteraciones en la normal diafonía celular, entre microglias y células neuronales cerebrales, pueden conducir a graves disturbios y enfermedades neurodegenerativas. En este contexto, hemos postulado que los procesos neuroinflamatorios son un factor crítico a desencadenar la cascada patológica que conduce a la degeneración neuronal. En nuestra teoría Neuroinmunomoduladora, señales de daños externos o internos activan las células microgliales, favoreciendo la producción de factores citotóxicos que inducen la degeneración neuronal. Estos factores activan la proteína-quinasas, que conducen a la hiperfosforilación de la proteína tau, y su consecuente oligomerización. Estos agregados de tau liberados al medio extracelular, al activar a la célula microglial, provocarían un mecanismo de retroalimentación positiva favoreciendo la neurodegeneración. Hoy en día, compuestos de origen natural con una fuerte actividad anti-inflamatoria, capaces de cruzar la barrera hematoencefálica del cerebro, aparecen como candidatos para la prevención y el tratamiento de trastornos neurodegenerativos tales como la enfermedad de Alzheimer.

Parâmetros clínicos e moleculares podem ser usados como biomarcadores das doenças peri-implantares e periodontais? / Can clinical and molecular parameters be used as biomarker of peri-implant and periodontally affected sites? - a literature review

O objetivo deste trabalho foi realizar uma revisão da literatura sobre os parâmetros clínicos e moleculares como marcadores de doenças peri-implantares e periodontais. Uma busca eletrônica foi realizada entre os anos de 2000 e 2014, apenas em periódicos de língua inglesa, utilizando-se as palavras-chave “dental implants”, “interleukin-1” e “peri-implantitis” no site Medline/PubMed. Parâmetros clínicos – sangramento (SS), profundidade de sondagem (PS), índice gengival (IG) e índice de placa (IP) –, bem como as diversas citocinas pró-inflamatórias, foram analisados em estudos com seres humanos com mucosite e, ou peri-implantite. Foram recuperados 325 artigos com 45 destes inicialmente selecionados e, após a avaliação dos textos completos, foram incluídos 20 trabalhos. Pôde-se concluir que: 1) As citocinas pró-inflamatórias são utilizadas como marcadores das doenças peri-implantar e periodontal; 2) Os trabalhos apresentam diversos parâmetros clínicos e moleculares como meio de diagnóstico; 3) Estes achados clínicos mostram a doença peri-implantar apenas em seu estágio atual; 4) Os resultados são variados pela natureza transversal dos estudos e das populações; e 5) Novas avaliações deverão ser feitas com o intuito de reforçar estas informações e torná-las úteis à prática clínica diária.

The aim of this paper was to make a literature review on the clinical and molecular parameters as biomarkers of the peri-implant and periodontal diseases. An electronic search was made between the 2000 and 2014 in the English language journals at the Medline/PubMed site. Clinical parameters - bleeding on probing (BoP), pocket depth (PD), gingival index (GI), and the plaque index (PI), as well as proinfl ammatory cytokines were analyzed in human models with mucositis and peri-implantitis. Overall, 325 references were retrieved, 45 initially selected, but only 20 papers included. it can be concluded that: 1) the proinfl ammatory cytokines are used as markers of peri-implant and periodontal diseases; 2) several clinical and molecular parameters were used for diagnostic purposes; 3) The clinical fi ndings show the peri-implant disease on its early stages; 4) The results varied due to its cross-sectional nature and targeted populations; and 5) New evaluations need to be made with the aim to reinforce these information and make them useful at the clinical practice.

[Immunometabolism of exercise and sedentary lifestyle]. / El inmunometabolismo del ejercicio físico y la vida sedentaria.

Sedentary lifestyle leads to the accumulation of visceral fat. This is accompanied by the infiltration of immune cells with pro-inflammatory characteristics in adipose tissue, causing an increased release of cytokines and generating a low-grade inflammatory state. It has been associated with the development of insulin resistance, atherosclerosis, neurodegeneration, and development of tumors. Exercise can be used as a treatment to improve symptoms of many of these conditions because it promotes an anti-inflammatory effect. In this review we analyze the pro-inflammatory factors present in obesity and the induction of antiinflammatory factors that occur with exercise.

La vida sedentaria induce la acumulación de grasa visceral, que se acompaña de la infiltración de células inmunitarias con características proinflamatorias en el tejido adiposo, que ocasiona mayor liberación de citocinas y genera un estado inflamatorio de bajo grado, éste se ha asociado con resistencia a la insulina, aterosclerosis, neurodegeneración y tumores. El ejercicio físico puede indicarse como tratamiento para disminuir los síntomas de muchas afecciones porque promueve un estado antiinflamatorio. En esta revisión se analizarán los factores proinflamatorios que coexisten en la obesidad, y la inducción de factores antiinflamatorios que se originan con el ejercicio físico.

Metotrexate como inductor en la producción de citocinas proinflamatorias en células epiteliales / Methotrexate as inducer of proinflammatory cytokines by epithelial cells

El metotrexate (MTX) es uno de los medicamentos frecuentemente utilizados en el cáncer infantil señalándose además, como agente citotóxico de la mucosa bucal, que puede desencadenar el proceso inflamatorio e incremento de la vascularidad en los tejidos epiteliales durante las fases iniciales de la mucositis oral. El presente trabajo tiene como objetivo determinar la producción de citocinas proinflamatorias IL-1b, IL-6 y TNF-a en cultivos de células epiteliales tratadas con MTX. Se realizaron cultivos de células epiteliales de laringe humana obtenidas de la línea celular Hep-2, con diferentes dosis de MTX en distintos tiempos de incubación, y a su vez se analizó la citotoxicidad del fármaco mediante el ensayo colorimétrico, el cual se basa en la reducción metabólica del bromuro de 3-(4,5- dimetiltiazol-2-ilo)-2,5-difeniltetrazol (MTT), y la producción de citocinas proinflamatorias mediante el ensayo inmuno enzimático indirecto (ELISA). En cuanto a los resultados se observó, que los cultivos de células Hep-2 presentaron aumento en la producción de las citocinas proinflamatorias a las 72 horas al utilizar las dosis de 0.32µM MTX. Estos resultados sugieren que la dosis y el tiempo de exposición del MTX alteran la fisiología de las células epiteliales humanas, lo cual podrían desempeñar un papel importante durante las fases de iniciación y de desarrollo de la mucositis oral.

Methotrexate (MTX), a drug commonly used in childhood cancer, has also been indicated as a cytotoxic agent of the oral mucosa, which can trigger the inflammatory process and increase the vascularity of epithelial tissues during the early stages of oral mucositis. The aim of this study was to determine the production of proinflammatory cytokines IL-1b, IL-6 y TNF-a in epithelial cell cultures treated with MTX. Epithelial cells of human larynx, obtained from the cell line Hep-2, were cultured with different doses of MTX during different incubation times. The drug cytotoxicity was analyzed by means of the colorimetric test, which is based on the metabolic reduction of the bromide of 3-(4, 5-dimetiltiazol-2-ilo)-2,5-difeniltetrazol (MTT); and the proinflammatory cytokines production by the test enzyme-linked immunosorbent assay (ELISA). Cultures of HEp-2 cells showed increased production of proinflammatory cytokines at 72 hours with 0.32µM of MTX. These results suggest that depending on the dose and exposure time, MTX alters the physiology of human epithelial cells, which may play an important role during the phases of initiation and development of oral mucositis.

Dendrimers as anti-inflammatory agents

Dendrimers constitute an intriguing class of macromolecules which find applications in a variety of areas including biology. These hyperbranched macromolecules with tailored backbone and surface groups have been extensively investigated as nanocarriers for gene and drug delivery, by molecular encapsulation or covalent conjugation. Dendrimers have provided an excellent platform to develop multivalent and multifunctional nanoconjugates incorporating a variety of functional groups including drugs which are known to be anti-inflammatory agents. Recently, dendrimers have been shown to possess anti-inflammatory properties themselves. This unexpected and intriguing discovery has provided an additional impetus in designing novel active pharmaceutical agents. In this review, we highlight some of the recent developments in the field of dendrimers as nanoscale anti-inflammatory agents.

Dendrímeros constituem uma classe intrigante de macromoléculas que apresentam aplicações em diversas áreas incluindo biologia. Essas macromoléculas extremamente ramificadas com esqueleto planejado e grupos de superfície foram extensivamente investigadas como nanotransportadores de genes e de fármacos, por encapsulamento molecular ou conjugação covalente. Dendrímeros têm proporcionado uma plataforma excelente de desenvolvimento nanoconjugados multivalentes e multifuncionais incorporando uma variedade de grupos funcionais, incluindo fármacos que são conhecidos por atuarem agentes antiinflamatórios. Recentemente, os dendrímeros mostraram propriedades antiinflamatórias. Esta inesperada e intrigante descoberta tem proporcionado um impulso adicional no planejamento de novos agente farmacêuticos ativos. Nesta revisão, nós destacamos alguns dos desenvolvimentos recentes no campo dos dendrímeros como agentes antiinflamatórios em nanoescala.