Signaling pathways associated with the expression of inflammatory mediators activated during the course of two models of experimental periodontitis

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Th1 polarized response induced by intramuscular DNA-HSP65 immunization is preserved in experimental atherosclerosis

We previously reported that a DNA vaccine constructed with the heat shock protein (HSP65) gene from Mycobacterium leprae (DNA-HSP65) was protective and also therapeutic in experimental tuberculosis. By the intramuscular route, this vaccine elicited a predominant Th1 response that was consistent with its protective efficacy against tuberculosis. It has been suggested that the immune response to Hsp60/65 may be the link between exposure to microorganisms and increased cardiovascular risk. Additionally, the high cholesterol levels found in atherosclerosis could modulate host immunity. In this context, we evaluated if an atherogenic diet could modulate the immune response induced by the DNA-HSP65 vaccine. C57BL/6 mice (4-6 animals per group) were initially submitted to a protocol of atherosclerosis induction and then immunized by the intramuscular or intradermal route with 4 doses of 100 µg DNA-HSP65. on day 150 (15 days after the last immunization), the animals were sacrificed and antibodies and cytokines were determined. Vaccination by the intramuscular route induced high levels of anti-Hsp65 IgG2a antibodies, but not anti-Hsp65 IgG1 antibodies and a significant production of IL-6, IFN-g and IL-10, but not IL-5, indicating a Th1 profile. Immunization by the intradermal route triggered a mixed pattern (Th1/Th2) characterized by synthesis of anti-Hsp65 IgG2a and IgG1 antibodies and production of high levels of IL-5, IL-6, IL-10, and IFN-g. These results indicate that experimentally induced atherosclerosis did not affect the ability of DNA-HSP65 to induce a predominant Th1 response that is potentially protective against tuberculosis.

Cell localization of the anti-inflammatory protein annexin 1 during experimental inflammatory response

The localization of the glucocorticoid-inducible protein annexin 1 (ANX-1) in leukocytes during the process of experimental inflammation has been studied using immunocytochemistry. ANX-1 immunoreactivity was detected in extravasated neutrophils and eosinophils as well as in resident tissue mast cells. Following injection of carrageenin, the mesenteric tissue was highly inflamed with large presence of leukocytes (predominantly neutrophils with a small percentage of eosinophils) adherent to post-capillary venules and extravasated in the perivascular tissue. ANX-1 immunoreactivity was detected in the cytosol of neutrophils and eosinophils mainly associated with granules and/or vesicles. A good degree of localization in the endosomes was observed in the neutrophils, In both cell types, some ANX-1 immunoreactivity in the nucleus and in the plasma membrane was also detected. Resident mast cells were also activated. Mast cells were positive for ANX-1, without apparent changes in protein content in relation to their activation status. Degranulated mast cells still presented ANX-1 associated with the granule matrix. In conclusion, this study demonstrated the presence of ANX-1 in leukocytes that play a central role in the host inflammatory response. These are the extravasating polymorphonuclear cells, or the resident mast cells. These data provide morphological support to the notion that endogenous and exogenous ANX-1 are able to modulate the reactivity of these cell types, and more generally, of the experimental inflammatory reaction.

An experimental assessment of landscape configuration effects on frog and toad abundance and diversity in tropical agro-savannah landscapes of southeastern Brazil

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Experimental paracoccidioidomycosis in hamster: transmission electron microscopy of inoculation site lesion

Estudou-se sequencialmente, à microscopia eletrônica de transmissão, a interação entre Paracoccidioides brasiliensis (Pb) e células inflamatórias em hamsters inoculados por via intratesticular. Seis horas após inoculações havia predominância de neutrófilos, estando presentes algumas células mononucleares e eosinófilos. Os neutrófilos foram progressivamente substituídos por células mononucleares. Fungos viáveis apresentavam-se fagocitados ou circunscritos por células inflamatórias, geralmente com ampla interface hospedeiro-parasita. Fungos mortos ou degenerados eram acompanhados de interfase estreita. A camada externa da parede do Pb era às vezes quebrada quando em contacto com neutrófilos, em vários pontos, sendo os fragmentos dessa parede descamados e fagocitados. Células fúngicas pequenas com um único núcleo se relacionavam com ampla interfase enquanto as células maiores e multinucleadas apresentavam paredes irregulares, por vezes, contendo lomasoma e/ou estrutura semelhante à mielina. Diferentes padrões de interação do Pb com células do hospedeiro podem ser decorrentes do a fluxo de células inflamatórias funcionalmente diferentes ao local de inoculação ou à idade dos fungos ou ambos os fatores.

Life cycle and host specificity of Amblyomma triste (Acari : Ixodidae) under laboratory conditions

We report biological data of two generations of Amblyomma triste in laboratory and compared the suitability of different host species. Infestations by larval and nymphal stages were performed on guinea pigs (Cavia porcellus), chickens (Gallus gallus), rats (Rattus norvegicus), rabbits (Oryctolagus cuniculus), wild mice (Calomys callosus), dogs (Canis familiaris) and capybaras (Hydrochaeris hydrochaeris). Infestations by adult ticks were performed on dogs, capybaras and rabbits. Tick developmental periods were observed in an incubator at 27degreesC and RH 90%. Guinea pigs were the most suitable hosts for larvae and nymphs, followed by chickens. The remaining host species were less suitable for immature ticks as fewer engorged ticks were recovered from them. Mean larval feeding periods varied from 3.8 to 4.7 d between different host species. Mean larval premolt periods ranged from 8.9 to 10.4 d. Nymphal mean feeding periods varied from 4.2 to 6.2 d for ticks fed on different host species. Premolt period of male nymphs (mean: 15.4 d) was significantly longer than that of female nymphs (14.7 d). Female nymphs were significantly heavier than male nymphs. The overall sex ratio of the adult ticks emerged from nymphs was 0.9:1 (M:F). Capybaras were the most suitable host for the tick adult stage as significantly more engorged females were recovered from them and these females were significantly heavier than those recovered from dogs or rabbits. The life cycle of A. triste in laboratory could be completed in an average period of 155 d. The potential role of guinea pigs, birds and capybaras, as hosts for A. triste in nature, is discussed.

Ticks (Acari : Ixodidae) parasitizing humans in an Atlantic rainforest reserve of Southeastern Brazil with notes on host suitability

While conducting projects on ticks from deer and on tick ecology in animal trails in an Atlantic rainforest reserve in Southeastern Brazil, researchers of our group were bitten by ticks several times. Some of these episodes were recorded. Three species of adult ticks attached to humans: Amblyomma brasiliense Aragauo, Amblyomma incisum Neumann, and Amblyomma ovale Koch. Eight nymphal attachments with engorgement on humans were recorded. From these, six molted to adults of A. incisum, one to an adult of A. brasiliense, and one had an anomalous molting, therefore the adult tick could not be properly identified. Local reactions to tick attachment varied among individual hosts from almost imperceptible to intense. Especially itching, but hyperemia and swelling as well, were prominent features of the reaction. Overall it can be affirmed that human beings can be a physiologically suitable host species for ticks in the Atlantic rainforest and that itching was an important if not the major component of the resistance to tick bite.

Actinobacillus actinomycetemcomitans lipopolysaccharide-mediated experimental bone loss model for aggressive periodontitis

Background: Bacterial constituents, such as Gram-negative derived lipopolysaccharide (LPS), can initiate inflammatory bone loss through induction of host-derived inflammatory cytokines. The aim of this study was to establish a model of aggressive inflammatory alveolar bone loss in rats using LPS derived from the periodontal pathogen Actinobacillus actinomycetemcomitans.Methods: Eighteen female Sprague-Dawley rats were divided into LPS test (N = 12) and saline control (N = 6) groups. All artimals received injections to the palatal molar gingiva three times per week for 8 weeks. At 8 weeks, linear and volumetric alveolar bone loss was measured by micro-computed tomography (mu CT). The prevalence of inflammatory infiltrate, proinflammatory cytokines, and osteoclasts was assessed from hematoxylin and eosin, immunohistochemical, or tartrate-resistant acid phosphatase (TRAP)-stained sections. Statistical analysis was performed.Results: A. actinomycetemcomitans LPS induced severe bone loss over 8 weeks, whereas control groups were unchanged. Linear and volumetric analysis of maxillae by mu CT indicated significant loss of bone with LPS, administration. Histologic examination revealed increased inflammatory infiltrate, significantly increased immunostaining for interleukin IL-6 and -1 beta and tumor necrosis factor-alpha, and more TRAP-positive osteoclasts in the LPS group compared to controls.Conclusion: Oral injections of LPS derived from the periodontal pathogen A. actinomycetemcomitans can induce severe alveolar bone loss and proinflammatory cytokine production in rats by 8 weeks.

Infecção experimental pelo Encephalitozoon cuniculi em camundongos imunossuprimidos com dexametasona

Objective Microsporidian Encephalitozoon cuniculi has been recognized as an opportunistic pathogen in immunosuppressed individuals, such as AIDS patients. The objective of the study was to develop pharmacologically immunosuppressed animals as a model of the natural occurring E. cuniculi infection.Methods Distint groups of adult Balb-C mice were immunosuppressed with different doses of dexamethasone (Dx, 3 or 5 mg/kg/day, intraperitoneal route - IP) and inoculated with E. cuniculi spores by IP route intraperitoneally. Control groups (inoculated animals but non-immunosuppressed and non-inoculated animals but immunosuppressed) were also used. The spores of E. cuniculi were previously cultivated in MDCK cells. The animals were sacrificed and necropsied at 7, 14, 21, 28 and 35 days post-inoculation. Tissue fragments were collected and processed for light microscopy studies, using Gram-chromotrope and hematoxylin-eosin staining techniques.Results In all immunosupressed and inoculated inoculated immunosuppressed mice,specially in those that received 5 mg/kg/day of dexamethasone, the most prominent necropsy findings were hepatomegaly and splenomegaly. The experimental inoculation resulted in a disseminated non-lethal infection, characterized by granulomatous lesions in several organs (liver lungs, kidneys, gut and brain) but notably in the hepatic tissue. Spores of E. cuniculi were only seen in few animals treated with 5 mg/kg/day of Dx at 35 days post-infection.Conclusions Microsporidiosis in Dx-immunosuppressed mice provides a useful model for studies of the microsporidial infection, resembling that one naturally occurring in immunodeficient individuals with AIDS.

Host-guest complexation of a nitroheterocyclic compound with cyclodextrins: a spectrofluorimetric and molecular modeling study

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Rabbit antibody responses to experimental infestation with Dermatobia hominis.

The three larval stages of Dermatobia hominis (Linnaeus) have been evaluated for their immunogenicity by ELISA and immunodiffusion (ID) using sera from experimentally infested rabbits. During a primary infestation, first instar D. hominis were found to cause most reaction and allowed the earliest diagnosis by ELISA. An inhibition of the antibody response against second and third instars was observed. The inhibition disappeared after departure of the larvae from the host. In experimentally immunized hosts the antibody response, following challenge, was highest against second and third instar antigens. Antibody remained elevated during the infestation but fell immediately after the larvae had left the host.

Experimental analysis of the effect of the structure area on the PZT-based SHM systems

The structural health monitoring (SHM) systems based on electromechanical (E/M) impedance technique have been widely investigated. Although many studies indicate the reliability of this technique, some practical considerations still have to be considered in real applications. This paper presents an experimental analysis of the effect of the structure area on the system's performance. The results indicate that the sensitivity of the system to detect damage decreases significantly when the host structure has large cross-section area. Copyright © 2009 by ASME.

Osso esponjoso liofilizado de cão utilizado como enxerto puro e associado a plasma rico em plaquetas ou medula óssea em falhas ósseas induzidas em coelhos: estudo experimental

Pós-graduação em Cirurgia Veterinária - FCAV

Efeito da suplementação nutricional com cogumelos A. blazei sobre a infecção experimental murina aguda por Paracoccidioides brasiliensis

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Ação imunomodulatória da β-glucana e Saccharomyces cerevisiae em ratos (Rattus rattus) da linhagem Wistar imunossuprimidos pela administração experimental de dexametasona

Pós-graduação em Medicina Veterinária - FCAV