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Characterisation of blood myeloid dendritic cells in mannose binding lectin-sufficient and mannose binding lectin-deficient individualsMelinda Dean Unknown Date (has links)
Mannose binding lectin (MBL) belongs to the collectin family of soluble pattern recognition molecules that elicit diverse biologic activities. Via multiple carbohydrate-recognition domains (CRD), MBL binds to mannose and N-acetyl-glucosamine oligosaccharides present on the surface of bacteria, fungi and yeast. Following pathogen recognition, MBL activates the complement system via MBL associated serine proteases in a manner independent of antibody and C1 complex. Deficiency in function and level of MBL is found in 25% of otherwise apparently healthy individuals, representing the most prevalent innate immune deficiency. MBL deficiency is a risk factor for the development of infections in humans and mice. The role of MBL as a modulator of infection is complex. MBL deficiency may influence proinflammatory cytokine production, expression of leukocyte adhesion molecules, or vascular damage, during the course of infection. Given that dendritic cells (DC) are antigen presenting cells (APC) with potent capacity to respond to microbial stimulation, I hypothesized that MBL deficiency may be reflected in DC functions associated with microbial stimulation. Initially, I investigated the association of MBL with human immune cells and demonstrated that in both MBL-Sufficient (MBL-S) and MBL-Deficient (MBL-D) individuals, MBL was particularly associated with monocytes. RT-PCR analysis demonstrated MBL was not transcribed by monocytes or other immune cells investigated (T, B, and NK cells, CD11c+DC, immature monocyte derived DC [MoDC], LPS matured MoDC, and granulocytes), suggesting MBL association with the cell surface may be via an adapter or co-receptor. Magnetically separated monocytes but not MoDC bound exogenous purified human plasma MBL (hpMBL). Addition of hpMBL (5 -15 µg/mL) did not induce MoDC activation, and MBL added together with lipopolysaccharide (LPS) did not induce MoDC activation above the level induced by LPS only. In the second part of this study, I used the particulate MBL ligand zymosan (Zy) as a pathogenic stimulus in a whole blood model to gain a greater understanding of the consequences of MBL deficiency. I compared surface phenotype, inflammatory cytokine production and antigen presenting capacity of blood myeloid (M)DC of MBL-D and MBL-S individuals following stimulation with Zy and MBL opsonised Zy (MBL-Zy). Blood MDC in MBL-D individuals, unlike their counterpart in MBL-S individuals, displayed unique functional characteristics, including higher production of proinflammatory cytokines IL-6 and TNF-, but poor capacity for allo-T cell effector cell induction. It appeared that stimulation with MBL-Zy reduced elevated production of IL-6 but not TNF- by blood MDC in MBL-D individuals. In the third part, expression microarray analysis was utilised to provide broad information on the genes and potential signalling pathways involved in the MDC responses in MBL-D and MBL-S individuals following stimulation with Zy and MBL-Zy. MBL-S individuals demonstrated greater capacity to induce T cell and NK cell signalling pathways than MBL-D individuals. Further, MBL acted as a regulator of important inflammatory molecules, namely T-cell receptor zeta (CD247), IFN-γ and perforin 1. The data presented in this study provides novel information on blood MDC function in MBL-S and MBL-D individuals in response to pathogen stimulation, and provided insight into mechanisms involved in the increased frequency of infection observed in MBL-D individuals.
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Influência da suplementaçao nutricional com glutamina na translocação bacteriana, endotoxemia, liberação de fator de necrose tumoral alfa, em ratos Wistar submetidos r induçao inflamatória sistêmica pelo ZymosanAndrade Neto, Jose Luiz de, 1958- January 2003 (has links)
Orientador: Joao Carlos Domingues Repka / Tese (doutorado) - Universidade Federal do Paraná, Setor de Ciencias da Saúde, Programa de Pós-graduação em Medicina Interna / Inclui bibliografia / Resumo: Este trabalho é o resultado do estudo da prevalência do vírus tipo 1 da imunodeficiência humana (HIV-1) com identificação dos prováveis fatores de risco e co-prevalência do HIV-1 com o antígeno de superfície do vírus da hepatite B (HBsAg) realizado em prostitutas na cidade portuária de Paranaguá, no Estado do Paraná, Brasil, no ano de 1 992. A obtenção dos dados foi possível através de questionário, havendo coleta de sangue para análise da presença dos anticorpos contra o HIV-1 e também da presença do antígeno de superfície do vírus da hepatite B. A freqüência da soropositividade para o HIV-1 foi de 4,5% e do HBsAg foi de 3,0%. A presença do HBsAg está estatisticamente relacionada com a postividade do HIV-1. O fato de não usar preservativo por não ver importância e o contato com parceiros bissexuais se mostraram como fatores de risco estatisticamente significativos para a infecção pelo HIV-1 / Abstract: This work is the result of a study about the prevalence of the type 1 virus of human immunodeficiency (HIV-1) along with the identification of the probable risk factors and co-prevalence of the HIV-1 with the surface antigen of hepatitis B virus (HBsAg), which took place in the city of Paranagua, which is a harbour, in the State of Parana, in Brazil, with prostitutes in the year of 1992. The collecting of the data was done through questionaires and blood samples to identify the presence of antibody of the HIV-1 and also the presence of the of hepatitis B surface antigen. The soropositivity frequency of the HIV-1 was 4,5% and the HBsAg was 3.0%. The presence of HBsAg is statistically related to the HIV-1 presence. The none use of condoms for not finding it necessary and also the sexual intercorse of bisexual partners are the major statistically proven risk factors for the HIV-1 infection. xiv
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Efeito antiinflamat?rio de fucana extra?da da alga Parda spatoglossum schroederii em modelos experimentais de Peritonite, choque n?o s?ptico e coliteSilva, Ana Katarina Andrade 17 May 2012 (has links)
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Previous issue date: 2012-05-17 / Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico / Fucans is a name used for sulfated polysaccharides, which is most
characteristic structure of the presence of sulfated L-fucose, are found in brown
seaweed (Phaeophyceae) and echinoderms (sea urchins and sea cucumbers).
These polysaccharides have been reported to possess anticoagulant, antitumor,
anti-viral, anti-proliferative and anti-inflammatory activities. Therefore, in
the present study was evaluate the effect of the fucan from the brown seaweed
Spatoglossum schroederii in models of peritonitis and non-septic shock induced
by zymosan, as well as in a murine model of colitis induces by DSS. So, the
mice treatment by intravenous route with the fucan was able to reduce the
exudate formation and the cell migration in the model of acute peritonitis
induced by zymosan during the kinetic of 6, 24 and 48 hours. Similarly, in the
model of non-septic shock induced by zymosan the fucan demonstrated a
protector effect to inhibited the cellular migration to the peritoneo, to decrease
the levels of IL-6 in the serum and in the peritoneal exudate, to attenuate the
lose of weight in the mice; beside to reduce the serum levels of hepatic
transaminases and as well as the liver injury. In the model of murine colitis, the
treatment with the fucan reduced the lose of weight of the animals, decreased
the levels of IL-17 and IFN- produced in the gut and decrease the intestinal
lesion induced by DSS. In conclusion, the fucan used in this study presented a
significant protector effect in the murine models of inflammation / Fucana ? uma denomina??o utilizada para polissacar?deos sulfatados,
que tem como caracter?stica estrutural mais marcante a presen?a de L-fucose
sulfatada, sendo encontrados em algas pardas (Phaeophyceae) e em
equinodermos (ouri?os e pepinos do mar). Esses polissacar?deos tem sido
descritos por possuir atividade anticoagulante, anti-tumoral, anti-viral, antiproliferativa
e anti-inflamat?ria. Portanto, no presente estudo foi avaliado o
efeito da fucana da alga parda Spatoglossum schroederii em modelos de
peritonite e choque n?o s?ptico induzido por zimosan, bem como em um
modelo murino de colite induzida por DSS. Dessa forma, o tratamento de
camundongos pela via intravenosa com a fucana foi capaz de reduzir a
forma??o do exsudato e a migra??o celular no modelo de peritonite aguda
induzida por zimosan durante a cin?tica de 6, 24 e 48 horas. De maneira
semelhante, no modelo de choque n?o-s?ptico induzido por zimosan a fucana
demonstrou efeito protetor ao inibir a migra??o celular para o perit?nio, diminuir
os n?veis de IL-6 s?rico e no exsudato peritoneal, ao atenuar a perda de peso
do animais, al?m de reduzir os n?veis s?ricos das transaminases hep?ticas,
assim como a les?o no f?gado. No modelo murino de colite, o tratamento com a
fucana reduziu a perda de peso dos animais, diminuiu os n?veis de IL-17 e IFN-
produzidos no intestino e diminuiu a les?o intestinal ocasionada pela DSS.
Conclui-se ent?o, que a fucana usada nesse estudo apresentou efeito protetor
significativo diante dos modelos murinos de inflama??o
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Estudo do efeito da fraÃÃo nÃo dialisÃvel do lÃtex de Calotropis procera (ait) R. Br em modelos experimentais de inflamaÃÃo, com Ãnfase em artrite / Study of the effect of the non-dialyzable fraction of Calotropis procera (Ait) R. Br latex in experimental models of inflammation, with emphasis in arthritisCid Freitas Cavalcante 18 July 2007 (has links)
CoordenaÃÃo de AperfeiÃoamento de Pessoal de NÃvel Superior / Tendo em vista a prevalÃncia de doenÃas inflamatÃrias reumÃticas tais como a artrite reumatÃide em nosso meio e a necessidade constante de novas terapias alternativas Ãs drogas esterÃides e nÃo esteroidais, que apresentam tantos efeitos nocivos em uso prolongado, decidimos investigar a aÃÃo do lÃtex de Calotropis procera em modelos experimentais de inflamaÃÃo, com foco na artrite. O lÃtex dessa planta da famÃlia Asclepiadaceae, vastamente encontrada no Nordeste brasileiro, tem efeitos sabidamente antiinflamatÃrios em algumas situaÃÃes, notadamente a sua fraÃÃo protÃica majoritÃria, nÃo dialisÃvel (FNDL), menos tÃxica e desprovida de borracha. A fraÃÃo foi obtida a partir de um protocolo estabelecido por nossos colaboradores, que, em suma envolve vÃrias etapas de centrifugaÃÃo e diÃlise atà o produto final liofilizado. Utilizamos ratos Wistar machos com peso entre 230 e 280g e camundongos, Balb/c e Swiss, pesando entre 18 e 35g, em grupos de seis indivÃduos nos quais um controle, outro sham em que foi induzido o evento inflamatÃrio (artrite por Zy - AZy, artrite induzida por antÃgeno - AIA ou peritonite infecciosa - PI) e o experimental no qual foi tratado com FNDL. O grupo AZy foi submetido a teste de incapacitaÃÃo articular pelo modelo de tempo de suspensÃo da pata (TSP) e, apÃs sacrifÃcio, à contagem de cÃlulas total e diferencial no lavado articular, estudo histopatolÃgico da articulaÃÃo, avaliaÃÃo de permeabilidade vascular, dosagem de ADA e TNF-α. Os resultados foram expressos em mÃdia  e.p.m. e comparados em testes estatÃsticos ANOVA/Bonferroni, admitindo-se P<0.05 para significÃncia. A FNDL inibiu o TSP na AZy com efeito mÃximo na dose de 3mg/kg no t = 4h. Houve tambÃm reduÃÃo significativa da permeabilidade vascular, nos nÃveis sÃricos de TNF-α e de ADA, e melhora no quadro histopatolÃgico. TambÃm reduziu o influxo celular sinovial na AZy e AIA, sobretudo de neutrÃfilos. Na PI, inibiu a migraÃÃo celular e os nÃveis de ADA. Dessa forma, os resultados confirmaram o efeito antiinflamatÃrio da FNDL nestes modelos de artrite e peritonite e sugeriram um possÃvel mecanismo de aÃÃo relacionado a um aumento da adenosina, por conta da reduÃÃo dos nÃveis de ADA, ou por inibiÃÃo do TNF-α. Por fim procedemos a avaliaÃÃo da toxicidade da droga em tratamento crÃnico, constatando que a dose de 10mg/Kg apresenta bons efeitos farmacolÃgicos sem alteraÃÃes significativas nos parÃmetros selecionados / The objective of this dissertation was to investigate the effect of the latex from Calotropis procera in experimental models of inflammation, with special focus on arthritis. The plant belongs to Asclepiadaceae family and is largely found in Northeast of Brazil. The plant latex presents clear anti-inflammatory effects, which is kept by its non-dialyzable protein fraction (NDPF), a less toxic and rubber free fraction. NDPF was obtained according to a procedure established by our group, which, in short, involves several steps of centrifugation and dialysis. Male Wistar rats (230 â 280g) and Balb/c and Swiss male mice (18 â 35g) were organized in groups of 6 animals: a control group (no treatment), a sham group, in which only the inflammatory event was induced (arthritis by zymosan â AZy), an antigen-induced arthritis group (AIA), an infectious peritonitis group (IP) and an experimental group, treated with NDPF. The AZy group was submitted to an articular incapacitation test to measure the paw elevation time (PET, in seconds) and, after sacrifice, total and differential cell count in the intra-articular fluid, histopathological study of the articulation, vascular permeabilization, ADA and TNF-α assays. The results were expressed in means + S.E.M. and significative differences were accepted if p<0.05 (ANOVA/Bonferroni). NDPF inhibited PET in AZy with a maximum effect in the dose of 3mg/kg, at 4h after inflammation induction. There was also a significative reduction of vascular permeability, TNF-α and ADA serum levels and an improvement of the histopathological profile. NDPF also reduced the intra-articular influx of cells in AZy and AIA, especially of neutrophils. In IP, NDPF inhibited the cellular migration and the levels of ADA. So, the results confirmed the expected anti-inflammatory effect of NDPF in arthritis and peritonitis models and suggested a possible mechanism of action involving either an increase of adenosine concentration due to the reduction of ADA levels or an inhibition of TNF-α or both events. Finally, the NDPF chronical toxicity was evaluated confirming that the dose of 10mg/Kg presents well defined pharmacological effects without significant alterations on the animal parameters. The characteristics of NDPF open an interesting possibility for alternative therapy to substitute the toxic effects of steroidal and some non-steroidal drugs, used in rheumatoid diseases, so prevalent among our population
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Syk-dependent ERK Activation Regulates IL-2 and IL-10 Production by DC Stimulated with ZymosanSlack, Emma C., Robinson, Matthew J., Hernanz-Falcón, Patricia, Brown, Gordon D., Williams, David L., Schweighoffer, Edina, Tybulewicz, Victor L., Reis e Sousa, Caetano 01 June 2007 (has links)
Zymosan is a particulate yeast preparation that elicits high levels of IL-2 and IL-10 from dendritic cells (DC) and engages multiple innate receptors, including the Syk-coupled receptor dectin-1 and the MyD88-coupled receptor TLR2. Here, we show that induction of IL-2 and IL-10 by zymosan requires activation of ERK MAP kinase in murine DC. Surprisingly, ERK activation in response to zymosan is completely blocked in Syk-deficient DC and unaffected by MyD88 deficiency. Conversely, ERK activation in response to the TLR2 agonist Pam3Cys is completely MyD88 dependent and unaffected by Syk deficiency. The inability of TLR2 ligands in zymosan to couple to ERK may explain the Syk dependence of the IL-2 and IL-10 response in DC and emphasises the importance of Syk-coupled pattern recognition receptors such as dectin-1 in the detection of yeasts. Furthermore, the lack of receptor compensation observed here suggests that responses induced by complex innate stimuli cannot always be predicted by the signalling pathways downstream of individual receptors.
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Efeito protetor dos extratos de Ascaris suum e Coccidioides posadasii e da lectina da semente de Dioclea violacea na artrite por zymosan em ratos e camundongos / Effect protector of the Ascaris suum and Coccidioides posadasii extracts and lectin of the seeds Dioclea violacea in arthritis zymosan in rats and miceAna Karine Rocha de Melo Leite 15 January 2009 (has links)
CoordenaÃÃo de AperfeiÃoamento de Pessoal de NÃvel Superior / InteraÃÃes entre a resposta imune inata e adquirida participam na fisiopatologia de doenÃas auto-imunes. Embora infecÃÃes estejam associadas ao desenvolvimento de artrites crÃnicas, à possÃvel que exposiÃÃo a alguns germes, como helmintos e fungos, potencialmente influencie a prevalÃncia e/ou gravidade de doenÃas imunomediadas. Lectinas derivadas de plantas, por aÃÃo em receptores de resposta inata, podem modular inflamaÃÃo. NÃs investigamos o efeito dos extratos de Ascaris suum (AS) e de Coccidioides posadasii (CP) e de uma lectina isolada da Dioclea violacea (Dviol) na artrite induzida por zymosan (AZy). Ratos Wistar e camundongos Swiss receberam 1 mg ou 0,1 mg de zymosan intra-articular (i.art.), respectivamente. Grupos foram prÃ-tratados (30 min) com os extratos de AS (0,25 - 2,5 mg/animal; i.p ou p.o.), CP (1 - 100 Âg/animal; i.art., i.p. ou p.o.) ou Dviol (0,3 - 30 Âg i.art. ou 1 - 6 mg/Kg e.v.). Grupo nÃo-tratado (NT) recebeu Zy (i.art.) e veÃculo. Animais naive receberam apenas salina (i.art.) e veÃculo. A hipernocicepÃÃo foi avaliada atravÃs do teste de incapacitaÃÃo articular em s / 1min. O lavado articular foi usado para anÃlise do influxo celular (IC), nÃveis de nitrito e citocinas. A sinÃvia foi utilizada para histopatologia. O conteÃdo de glicosaminoglicanos (GAG) da cartilagem foi quantificado para medir dano estrutural. O extrato de AS, seja i.p. ou p.o., inibiu de forma dose-dependente a hipernocicepÃÃo e o IC na AZy em relaÃÃo ao grupo NT (P<0,01), bem como reverteu o dano articular avaliado pela quantificaÃÃo de GAG e a sinovite vista à histologia. A administraÃÃo do extrato de AS, reduziu significantemente os nÃveis de nitrito, inteleucina-1β (IL-1β) e IL-10, mas nÃo de fator de necrose tumoral alfa (TNF-α), em relaÃÃo ao NT. Em camundongos, o extrato de AS reduziu os nÃveis de IL-10, mas nÃo de IL-1β ou TNF-α. O tratamento com o extrato de CP, seja i.p. ou p.o., inibiu significantemente a hipernocicepÃÃo e o IC na AZy, em relaÃÃo ao NT, no entanto, nÃo reverteu a lesÃo articular medida pela quantidade de GAG e histologia. A administraÃÃo da Dviol, em animais naive promoveu IC significante, embora apenas a maior dose (30Âg) promoveu hipernocicepÃÃo. Na AZy, a injeÃÃo i.art. da Dviol reduziu o IC e hipernocicepÃÃo de forma dose-dependente, em relaÃÃo ao NT (P<0,01). A administraÃÃo da Dviol (i.v.) reduziu ambos hipernocicepÃÃo e IC na AZy, em relaÃÃo ao NT (P<0,01). O efeito da Dviol foi revertido quando essa lectina foi prÃ-incubada com manose 1 M. Os dados mostram que um extrato de AS promove melhora funcional e protege do dano estrutural na AZy, que sÃo associados com reduÃÃo na liberaÃÃo de NO e citocinas i.art. Esse efeito independe da espÃcie e ocorre por via oral. Um extrato do fungo CP tem aÃÃo anti-inflamatÃria na AZy. Uma lectina isolada da Dviol reduz IC e hipernocicepÃÃo na AZy, provavelmente por acoplamento a um receptor de manose. Em conjunto, os resultados mostram que substÃncias que agem em receptores de resposta inata modulam a inflamaÃÃo articular imunomediada. / The interactions between innate and acquired immune responses participate in the pathophysiology of the autoimmune diseases. Though infections are associate with the development of the chronic arthritis it is possible that exposure to some germs as helminthes and fungi influences potentially the prevalence and/or gravity of the immune diseases. Lectins derivate of the plants can modulate the inflammation by action in receptors of the innate response. We investigated the effect of extracts from Ascaris suum (AS), Coccidioides posadasii (CS) and a lectin isolated from Dioclea violacea (Dviol) in zymosan-induced arthritis (ZyA). Wistar rats and Swiss mice received 1 mg or 0.1 mg zymosan intra-articular (i.art.), respectively. Groups were pretreated (30 min) with AS (0.25 - 2.5 mg/animal; i.p. or p.o.) CP (1 - 100 Âg/animal; i.art. i.p. or p.o) or Dviol (0.3 - 30 Âg; i.art. or 1 - 6 mg/kg; i.v.). Non-treated group (NT) received Zy (i.art.) and the vehicle. Naive animals received just saline (i.art.) and the vehicle. The hypernociception was evaluated through articular incapacitation test in s/1min. The joint exudate was used for evaluation of cell influx (CI), nitrite and cytokine levels. The synovium was used for histopatology. The glycosaminoglycan (GAG) content of the cartilage was quantificated for the measured of the structural damage. The AS extract both i.p. and p.o. significantly and dose-dependently inhibited CI and hypernociception in ZyA as compared to NT (P<0.01) as well as reverted articular damage assessed by quantification of the GAG and by synovitis observed in the histology. The administration of the AS extract reduced significantly levels of nitrite, interleukin-1β (IL-1β) and IL-10, but not tumor necrosis factor alpha (TNF-α) as compared to NT. In mice, it reduced IL-10 but not IL-1β and TNF- α. The treatment with CP extract both i.p. and p.o. inhibited hypernociception and CI in ZyA as compared to NT, but not reverted articular injury measured by GAG and histology. The administration of the Dviol in naÃve animals promoted CI significant, though just the highest dose (30 g) promoted hypernociception. In ZyA, Dviol (i.art.) reduced the CI and hypernociception dose-dependently (P<0.01). The administration of Dviol (i.v.) significantly reduced both the hyperalgesia and CI in ZyA as compared to NT (P<0.01). The effect of the Dviol was reverted when it was pre-incubated with mannose (1M). The date show that AS extract promote functional improve and protect of the articular damage in ZyA that are associate with reduction of the NO and cytokine (i.art.) liberation. This effect is species independent and functions orally. An extract of the fungi CP has anti-inflammatory activity in ZyA. A lectin isolated of the Dviol reduces CI and hypernociception in ZyA probably by coupling the mannose receptor. Together the results show that substances that act in receptors of the innate response modulate the immunomediate articular inflammation.
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Zymosan-Induced Peritonitis: Effects on Cardiac Function, Temperature Regulation, Translocation of Bacteria, and Role of Dectin-1Monroe, Lizzie L., Armstrong, Michael G., Zhang, Xia, Hall, Jennifer V., Ozment, Tammy R., Li, Chuanfu, Williams, David L., Hoover, Donald B. 01 January 2016 (has links)
Zymosan-induced peritonitis is a model commonly used to study systemic inflammatory response syndrome and multiple organ dysfunction syndrome. However, effects of zymosan on cardiac function have not been reported. We evaluated cardiac responses to zymosan in mice and the role of β-Glucan and dectin-1 in mediating these responses. Temperature and cardiac function were evaluated before and after intraperitoneal (i.p.) injection of zymosan (100 or 500 mg/kg) or saline. Chronotropic and dromotropic functions were measured using electrocardiograms (ECGs) collected from conscious mice. Cardiac inotropic function was determined by echocardiography. High-dose zymosan caused a rapid and maintained hypothermia along with visual signs of illness. Baseline heart rate (HR) was unaffected but HR variability (HRV) increased, and there was a modest slowing of ventricular conduction. High-dose zymosan also caused prominent decreases in cardiac contractility at 4 and 24 h. Because zymosan is known to cause gastrointestinal tract pathology, peritoneal wash and blood samples were evaluated for bacteria at 24 h after zymosan or saline injection. Translocation of bacterial occurred in all zymosan-treated mice (n=3), and two had bacteremia. Purified β-Glucan (50 and 125 mg/kg, i.p.) had no effect on temperature or ECG parameters. However, deletion of dectin-1 modified the ECG responses to high-dose zymosan; slowing of ventricular conduction and the increase in HRV were eliminated but a marked bradycardia appeared at 24 h after zymosan treatment. Zymosan-treated dectin-1 knockout mice also showed hypothermia and visual signs of illness. Fecal samples from dectin-1 knockout mice contained more bacteria than wild types, but zymosan caused less translocation of bacteria. Collectively, these findings demonstrate that zymosan-induced systemic inflammation causes cardiac dysfunction in mice. The data suggest that dectin-1-dependent and -independent mechanisms are involved. Although zymosan treatment causes translocation of bacteria, this effect does not have a major role in the overall systemic response to zymosan.
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