Saccharomyces boulardii as therapeutic alternative in experimental giardiasis.

AIMS: The objective of this study was to evaluate the effect of treatment with the probiotic Saccharomyces boulardii with or without metronidazole in experimental giardiasis. METHODS AND RESULTS: The effect of treatment with S. boulardii with or without metronidazole on the intestinal mucosa, the antioxidant defence system and the parasitic load was determined in experimental giardiasis. Eight groups of animals with infection and/or treatment with the probiotic and/or drugs for 1 week after infection with Giardia lamblia were used. A reduction of approximately 90% in the parasitic load was observed in all the treated groups. Saccharomyces boulardii attenuated the damage caused by infection in the intestinal mucosa preserving its architecture and inhibiting the oxidative stress induced by parasite and metronidazole. CONCLUSIONS: Saccharomyces boulardii was effective alone or in combination with metronidazole in resolving already established G. lamblia infection. SIGNIFICANCE AND IMPACT OF THE STUDY: These results suggest the use of S. boulardii as an alternative treatment for giardiasis mainly in cases of resistance or intolerance to conventional treatment.

Weissella paramesenteroides WpK4 ameliorate the experimental amoebic colitis by increasing the expression of MUC-2 and the intestinal epithelial regeneration.

AIMS: This study evaluates the action of Weissella paramesenteroides WpK4 on amoebic colitis. METHODS AND RESULTS: Weissella paramesenteroides WpK4 was administered in Entamoeba dispar infected and noninfected mice and clinical parameters were evaluated. Following 7 days, the caeca were collected for histopathology, morphometry and immunohistochemical staining of MUC-2, CDC-47 and IgA. The treatment reduced diarrhoea and the presence of blood in the faeces and diminished the area of necrosis, also causing weight gain. Also, the addition of this bacterium enhanced the expression of the mucin (MUC-2). The reduction in necrosis and increased CDC-47 expression indicates significant epithelial regeneration. The negative correlation between CDC-47 and the necrosis area reveals that the bacterium favoured the recovery of the necrotic regions and the positive correlation found between the expression of MUC-2 and CDC-47 indicates that the epithelial regeneration also supports the synthesis of MUC-2. CONCLUSIONS: Weissella paramesenteroides WpK4 was able to increase the protection of the intestinal mucosa against experimental amoebic colitis through the increase of MUC-2 and epithelial regeneration. SIGNIFICANCE AND IMPACT OF THE STUDY: Weissella paramesenteroides WpK4 presents the potential to become a complementary tool in the treatment of amoebic colitis.

Probiotic effect of Bifidobacterium longum 51A and Weissella paramesenteroides WpK4 on gerbils infected with Giardia lamblia.

AIMS: The objective of this study was to assess the probiotic potential of genuine strains of Bifidobacterium longum 51A and Weissella paramesenteroides WpK4, in experimental giardiasis. METHODS AND RESULTS: The bacteria were administered orally to gerbils (Meriones unguiculatus) 10 days before oral infection with trophozoites of Giardia lamblia. After 7 days of infection, the animals were euthanized and portions of the duodenum were processed for histopathologic, histochemical and morphometric assessment. The height of the intestinal crypts and crypt/villi ratio were higher in infected groups (P < 0·05) than in noninfected groups. The area of mucus production was higher (P < 0·05) in infected animals pretreated with B. longum 51A than in other groups. The parasitic load of the animals that received both bacteria decreased significantly (P < 0·05) compared to the ones of the control group. CONCLUSIONS: Our results suggest a probiotic function of B. longum 51A and W. paramesenteroides WpK4 and may result in their use as a prophylactic and therapeutic alternative for promoting human and animal health. SIGNIFICANCE AND IMPACT OF THE STUDY: Bifidobacterium longum 51A and W. paramesenteroides WpK4 may constitute prophylactic alternatives, reversing the emergence of side effects and resistance observed in the conventional treatment of giardiasis.

Effect of probiotic Saccharomyces boulardii in experimental giardiasis.

The aim of the study was to assess the efficacy of Saccharomyces boulardii in experimental treatment of giardiasis and its impact on intestinal integrity and some functions of gerbils infected with Giardia lamblia. 28 gerbils (Meriones unguiculatus), aged 4-6 weeks, were divided into four groups: untreated and uninfected control (CT); infected with G. lamblia (IGL); treated with S. boulardii (SB); and infected with G. lamblia and treated with S. boulardii (ITSB). The SB and ITSB groups received S. boulardii 15 days prior to being infected with G. lamblia. The treatment continued until completion of the experiment (22nd day). The IGL and ITSB groups were gavage-inoculated with G. lamblia ensuring one-week infection. 4 h before euthanasia, all animals were gavaged with a solution containing diethylenetriamine-pentaacetic acid (DTPA) marked with technetium-99mTc DTPA to determine intestinal permeability. The small intestine was removed for histopathological, morphometric analysis and count of trophozoites adhered to the mucosa. The selected probiotic caused an approximate reduction of 70% of parasite load, which was determined by attached trophozoites (P<0.01) and immune-marked trophozoites (P<0.05). Treatment with S. boulardii (SB and ITSB groups) also increased the height of the intestinal villi and crypt depth compared to the CT and IGL groups (P<0.05). The area of mucus production and the number of goblet cells of the SB and ITSB groups were higher compared to the CT and IGL groups (P<0.01). The animals treated with S. boulardii also exhibited a significant increase of intraepithelial lymphocytes counts (P<0.01). There was no difference in the intestinal permeability between the groups studied. The efficacy of S. boulardii in reducing damages caused by Giardia was demonstrated, with an approximate reduction of 70% of the parasite load, suggesting its use as a coadjuvant in giardiasis treatment.

Healing activity of proteolytic fraction (P1G10) from Vasconcellea cundinamarcensis in a cutaneous wound excision model.

The proteolytic enzymes from Vasconcellea cundinamarcensis have demonstrated efficacy to accelerate healing of skin lesions. We report here the efficacy of the proteolytic fraction - P1G10 during repair of excisional wounds in rodent model and analyze possible mediators involved. Using 0.05% P1G10 we observed on day 3rd increased wound contraction accompanied by an increase in activated neutrophils and VEGF relative to the control. On day 7th neutrophils returned to normal levels, and at 0.01% P1G10, an increase in NAG activity used to monitor monocyte/macrophage, was observed. On the other hand, on day 7th, we observed a decrease in TGF-ß at 0.05% P1G10, accompanied by an increased transformation of the latent TGF-ß to its active form. Also, on day 7th a reduction in MMP-9 activity and the number of apoptotic cells was observed along with an increase in fibroblast levels. Morphometrically, it appears that treatment with P1G10 accelerates the decline of initial inflammatory phase and reduces some unwanted effects likely caused by remaining TGF-ß or MMPs, thus enhancing the quality of scar. Overall, these data suggest that the active proteolytic fraction P1G10 enhances the efficacy of repair in excisional cutaneous wounds.

Glycol methacrylate embedding for the histochemical study of the gastrointestinal tract of dogs naturally infected with Leishmania infantum.

In canine visceral leishmaniasis a diffuse chronic inflammatory exudate and an intense parasite load throughout the gastrointestinal tract has been previously reported. However, these studies did not allow a properly description of canine cellular morphology details. The aim of our study was to better characterize these cells in carrying out a qualitative and quantitative histological study in the gastrointestinal tract of dogs naturally infected with Leishmania infantum by examining gut tissues embedded in glycol methacrylate. Twelve infected adult dogs were classified in asymptomatic and symptomatic. Five uninfected dogs were used as controls. After necropsy, three samples of each gut segment, including esophagus, stomach, duodenum, jejunum, ileum, cecum, colon, and rectum were collected and fixed in Carnoy's solution for glycol methacrylate protocols. Sections were stained with hematoxylin-eosin, toluidine blue borate, and periodic acid-Schiff stain. Leishmania amastigotes were detected by immunohistochemistry employed in both glycol methacrylate and paraffin embedded tissues. The quantitative histological analysis showed higher numbers of plasma cells, lymphocytes and macrophages in lamina propria of all segments of GIT of infected dogs than controls. The parasite load was more intense and cecum and colon, independently of the clinical status of these dogs. Importantly, glycol methacrylate embedded tissue stained with toluidine blue borate clearly revealed mast cell morphology, even after mast cell degranulation. Infected dogs showed lower numbers of mast cells in all gut segments than did controls. Despite the glycol methacrylate (GMA) protocol requires more attention and care than the conventional paraffin processing, this embedding procedure proved to be especially suitable for the present histological study, where it allowed to preserve and observe cell morphology in fine detail.

Infection with CagA-positive Helicobacter pylori strain containing three EPIYA C phosphorylation sites is associated with more severe gastric lesions in experimentally infected Mongolian gerbils (Meriones unguiculatus).

Infection with Helicobacter pylori strains containing high number of EPIYA-C phosphorylation sites in the CagA is associated with significant gastritis and increased risk of developing pre-malignant gastric lesions and gastric carcinoma. However, these findings have not been reproduced in animal models yet. Therefore, we investigated the effect on the gastric mucosa of Mongolian gerbil (Meriones unguiculatus) infected with CagA-positive H. pylori strains exhibiting one or three EPIYA-C phosphorilation sites. Mongolian gerbils were inoculated with H. pylori clonal isolates containing one or three EPIYA-C phosphorylation sites. Control group was composed by uninfected animals challenged with Brucella broth alone. Gastric fragments were evaluated by the modified Sydney System and digital morphometry. Clonal relatedness between the isolates was considered by the identical RAPD-PCR profiles and sequencing of five housekeeping genes, vacA i/d region and of oipA. The other virulence markers were present in both isolates (vacA s1i1d1m1, iceA2, and intact dupA). CagA of both isolates was translocated and phosphorylated in AGS cells. After 45 days of infection, there was a significant increase in the number of inflammatory cells and in the area of the lamina propria in the infected animals, notably in those infected by the CagA-positive strain with three EPIYA-C phosphorylation sites. After six months of infection, a high number of EPIYA-C phosphorylation sites was associated with progressive increase in the intensity of gastritis and in the area of the lamina propria. Atrophy, intestinal metaplasia, and dysplasia were also observed more frequently in animals infected with the CagA-positive isolate with three EPIYA-C sites.  We conclude that infection with H. pylori strain carrying a high number of CagA EPIYA-C phosphorylation sites is associated with more severe gastric lesions in an animal model of H. pylori infection.

Identificação imuno-histoquímica de VEGF e IGF-1 em ovários de cadelas no anestro e estro / Immunohistochemical identification of VEGF and IGF-1 in ovaries of bitches on anestrus and estrus

Com o objetivo de verificar a presença de VEGF e IGF-1 nos ovários de cadelas, foram realizadas análises imuno-histoquímicas do estroma cortical; teca e granulosa de folículos secundários, terciários e terciários pré-ovulatórios luteinizados; e ovócitos de folículos primários, secundários e terciários de ovários de cinco cadelas em anestro (Anest) e cinco em estro (Est). A identificação das fases do ciclo estral foi realizada por citologia vaginal associada a dosagem plasmática de progesterona. Os ovários foram submetidos a tratamento imuno-histoquímico para identificação de VEGF (anticorpo primário PU 360-UP, Biogenex, USA; diluição 1:30) e IGF-1 (anticorpo primário PabCa, Gro-Pep, Austrália; diluição 1:100). Determinou-se um índice de imunomarcação (IM), para cada tecido avaliado, pela razão entre a área positivamente marcada dividida pela área total analisada. Para os ovócitos, verificou-se imunomarcação positiva ou negativa. As comparações de IM entre tecidos foram realizadas pelo teste de Wilcoxon (diferentes tecidos em mesmo grupo) ou Mann-Whitney (mesmo tecido entre diferentes grupos), todas no nível de 5% de significância. VEGF e IGF-1 foram identificados, de forma semelhante (P>0,05), em todas as estruturas avaliadas em ambos os grupos experimentais. Conclui-se que esses fatores de crescimento estão presentes em cadelas no anestro e estro, no estroma cortical ovariano, folículos em diferentes estádios de desenvolvimento e ovócitos.(AU)

In order to verify the presence of VEGF and IGF-1 in the ovaries of bitches, immunohistochemical analyzes of the cortical stroma; theca and granulosa of secondary, tertiary and tertiary luteinized preovulatory follicles; and oocytes of primary, secondary and tertiary follicles of ovaries from five bitches in anestrous (Anest) and five in estrus (Est) was performed. The identification of the phases of the estrous cycle was performed by vaginal cytology associated with the measurement of plasma progesterone. The ovaries were treated for immunohistochemical identification of VEGF (PU 360 primary antibody-UP, Biogenex, USA, dilution 1:30) and IGF-1 (primary antibody PabCa, Gro-Pep, Australia; 1:100 dilution). The immunostaining index (MI) was determined for each tissue by the ratio of positively marked area divided by total analyzed area. For oocytes immunostaining was determined as positive or negative. Comparisons of IM between tissues were performed with the Wilcoxon test (deferent tissues in the same group) or Mann-Whitney test (same tissue between different groups), all at 5% significance level. VEGF and IGF-1 have been similarly identified (P>0.05) in all structures evaluated in both groups. It is concluded that in bitches in estrus and anestrous these growth factors are present in ovary cortical stroma, follicles at different stages of development and oocytes.(AU)

Angiotensin-(1-7) attenuates airway remodelling and hyperresponsiveness in a model of chronic allergic lung inflammation.

BACKGROUND AND PURPOSE: A long-term imbalance between pro- and anti-inflammatory mediators leads to airway remodelling, which is strongly correlated to most of the symptoms, severity and progression of chronic lung inflammation. The Angiotensin-(1-7) [Ang-(1-7)]/Mas receptor axis of the renin-angiotensin system is associated with attenuation of acute and chronic inflammatory processes. In this study, we investigated the effects of Ang-(1-7) treatment in a model of chronic allergic lung inflammation. EXPERIMENTAL APPROACH: Mice were sensitized to ovalbumin (OVA; 4 injections over 42 days, 14 days apart) and were challenged three times per week (days 21-46). These mice received Ang-(1-7) (1 µg·h(-1) , s.c.) by osmotic mini-pumps, for the last 28 days. Histology and morphometric analysis were performed in left lung and right ventricle. Airway responsiveness to methacholine, analysis of Ang-(1-7) levels (RIA), collagen I and III (qRT-PCR), ERK1/2 and JNK (Western blotting), IgE (elisa), cytokines and chemokines (elisa multiplex), and immunohistochemistry for Mas receptors were performed. KEY RESULTS: Infusion of Ang-(1-7) in OVA-sensitized and challenged mice decreased inflammatory cell infiltration and collagen deposition in the airways and lung parenchyma, and prevented bronchial hyperresponsiveness. These effects were accompanied by decreased IgE and ERK1/2 phosphorylation, and decreased pro-inflammatory cytokines. Mas receptors were detected in the epithelium and bronchial smooth muscle, suggesting a site in the lung for the beneficial actions of Ang-(1-7). CONCLUSIONS AND IMPLICATIONS: Ang-(1-7) exerted beneficial attenuation of three major features of chronic asthma: lung inflammation, airway remodelling and hyperresponsiveness. Our results support an important protective role of Ang-(1-7) in lung inflammation.

A multivalent chimeric vaccine composed of Schistosoma mansoni SmTSP-2 and Sm29 was able to induce protection against infection in mice.

Schistosoma mansoni is a blood fluke parasite responsible for schistosomiasis. The best long-term strategy to control schistosomiasis is through immunization combined with drug treatment. In this study, we cloned, expressed and purified SmTSP-2 fused to the N- and C-terminal halves of Sm29 and tested these chimeras as vaccine candidates using an adjuvant approved to be used in humans. The results demonstrated that vaccination with SmTSP-2 fused to N- or C-terminus of Sm29-induced reduction in worm burden and liver pathology when compared to control animals. Additionally, we detected high levels of mouse-specific IgG, IgG1 and IgG2a against both chimeras and significant amounts of IFN-γ and TNF-α and no IL-4. Finally, studies with sera from patients resistant to infection and living in schistosomiasis endemic areas revealed high levels of specific IgG to both chimeras when compared to healthy individuals. In conclusion, SmTSP-2/Sm29 chimeras tested here induced partial protection against infection and might be a potential vaccine candidate.

AVE 0991, a non-peptide mimic of angiotensin-(1-7) effects, attenuates pulmonary remodelling in a model of chronic asthma.

BACKGROUND AND PURPOSE: AVE 0991 (AVE) is a non-peptide compound, mimic of the angiotensin (Ang)-(1-7) actions in many tissues and pathophysiological states. Here, we have investigated the effect of AVE on pulmonary remodelling in a murine model of ovalbumin (OVA)-induced chronic allergic lung inflammation. EXPERIMENTAL APPROACH: We used BALB/c mice (6-8 weeks old) and induced chronic allergic lung inflammation by OVA sensitization (20 µg·mouse(-1) , i.p., four times, 14 days apart) and OVA challenge (1%, nebulised during 30 min, three times per·week, for 4 weeks). Control and AVE groups were given saline i.p and challenged with saline. AVE treatment (1 mg·kg(-1) ·per day, s.c.) or saline (100 µL·kg(-1) ·per day, s.c.) was given during the challenge period. Mice were anaesthetized 72 h after the last challenge and blood and lungs collected. In some animals, primary bronchi were isolated to test contractile responses. Cytokines were evaluated in bronchoalveolar lavage (BAL) and lung homogenates. KEY RESULTS: Treatment with AVE of OVA sensitised and challenged mice attenuated the altered contractile response to carbachol in bronchial rings and reversed the increased airway wall and pulmonary vasculature thickness and right ventricular hypertrophy. Furthermore, AVE reduced IL-5 and increased IL-10 levels in the BAL, accompanied by decreased Ang II levels in lungs. CONCLUSIONS AND IMPLICATIONS: AVE treatment prevented pulmonary remodelling, inflammation and right ventricular hypertrophy in OVA mice, suggesting that Ang-(1-7) receptor agonists are a new possibility for the treatment of pulmonary remodelling induced by chronic asthma.

Carbon nanotubes functionalized with sodium hyaluronate restore bone repair in diabetic rat sockets.

OBJECTIVE: We evaluated the effects of sodium hyaluronate (HY) and carbon nanotubes functionalized with HY (HY-CNT) on bone repair in the tooth sockets of diabetic rats. MATERIALS AND METHODS: Diabetes was induced by streptozotocin (50 mg kg(-1) i.v.), and the sockets were divided into normal control, diabetic control, diabetic treated with HY (1%), and diabetic treated with HY-CNT (100 µg ml(-1)) groups. The sockets were analyzed according to the percentage of bone formation and the number of cell nuclei. RESULTS: The percentage of bone trabeculae was lower in diabetic control animals (11.16 ± 5.10% vs 41.92 ± 6.34% in normal animals) after 14 days. Treating diabetic animals with HY or HY-CNT significantly increased the percentage of neoformed trabeculae (HY: 29.43 ± 3.29%; HY-CNT: 36.90 ± 3.07%). Moreover, the sockets of diabetic animals had an increased number of cell nuclei and HY or HY-CNT reduced this parameter. CONCLUSION: Our results indicate that HY and HY-CNT restore bone repair in the tooth sockets of diabetic rats, suggesting that these biomaterials are potential adjuvant therapies for the management of diabetes.

Campomanesia lineatifolia Ruiz & Pav. as a gastroprotective agent.

ETHNOPHARMACOLOGICAL RELEVANCE: Campomanesia lineatifolia Ruiz and Pav. (Myrtaceae) is a native edible species found in the Amazon Rainforest, commonly known as gabiroba. In Brazil, Campomanesia species are frequently used in traditional medicine for gastrointestinal disorders. MATERIALS AND METHODS: The present study performed phytochemical analyses and determined both the in vitro antioxidant activity of the ethanolic extract of Campomanesia lineatifolia leaves (EEC) as well as its ethyl acetate fraction (EAFC). In this analysis, quercetin was used as a positive control. Gastroprotective activity was also investigated at different oral doses in two experimental models in rats - gastric lesion induced by ethanol and gastric lesion induced by indomethacin. In this analysis, cimetidine and sucralfate were used as positive controls. The area of gastric lesion underwent macroscopic and histomorphometric evaluations, while the mucus content was estimated by applying the periodic acid-Schiff stain. Oral acute toxicity was also assessed. RESULTS: Phytochemical studies revealed the presence of flavonoids and tannins. Catechin and quercitrin were isolated by bioguided chromatographic fractionation of EAFC. EEC and EAFC presented in vitro antioxidant activity. The oral administration of EEC and EAFC at doses 100-400 mg/kg (ethanol model) and at doses of 400-1200 mg/kg (indomethacin model) proved to be effective in preventing gastric ulcerations in rats. Pretreatment with EAFC (400mg/kg) significantly increased the gastric mucus content in the ethanol model. No animals died during the acute oral toxicology test. CONCLUSIONS: Results confirm the Brazilian ethnopharmacological use of Campomanesia lineatifolia as a gastroprotective agent and indicate that the anti-ulcer effect is most likely mediated by scavenging free radicals due to the polyphenol content and, at least in part, by increasing the mucus secretion and the mucosal defense. In addition, EEC and EAFC were found to be safe when applied to a 2000 mg/kg single oral dose.

The role of iNOS and PHOX in periapical bone resorption.

Nitric oxide (NO) and reactive oxygen species (ROS) are key molecules in resistance to pathogens. Little is known about their role in pathogenesis of periapical lesions. To address this issue, we induced periapical lesions in mice lacking nitric oxide synthase (iNOS(-/-)) or phagocyte oxidase (PHOX(-/-)). iNOS(-/-) mice expressed higher levels of IL-1ß, TNF-α, RANK, RANKL, and MCP-1 than C57BL/6 and PHOX(-/-). Apical thickening of the periodontal ligament was also greater in iNOS(-/-) compared with other groups. Interestingly, ROS production did not interfere in periapical lesion progression, but seemed to be essential for the appearance of multinucleated TRAP-positive cells. Thus, periapical lesion progression in iNOS(-/-) was associated with an imbalance of pro-inflammatory cytokines (IL-1ß and TNF-α), bone-resorptive modulators (RANK and RANKL), and MCP-1. We conclude that NO, but not ROS, controls progression of bone resorption in a murine experimental model of apical periodontitis.

Immunization with newly transformed Schistosoma mansoni schistosomula tegument elicits tegument damage, reduction in egg and parasite burden.

The surface of the schistosomula is an important target for host immune system attack because the tegument represents the interface between host and parasite and thus is a potential candidate for the development of new intervention strategies. In this study, we evaluated the ability of schistosomula tegument (Smteg) to induce protection in mice. Immunization of mice with Smteg together with Freund adjuvant induced a Th1 type of immune response associated with a significant reduction in worm burden (43-48%), eggs trapped in the liver (65%), eggs eliminated in the faeces (59-60%) and granuloma number (41%). Lastly, during an in vitro study, worms from mice immunized with Smteg showed damage in the adult worm tegument and impaired egg laying.

Histopathological and immunohistochemical study of the hepatic lesions experimentally induced by Entamoeba dispar.

The sequence of hepatic necrotic-inflammatory events produced by Entamoeba dispar are originally described in this work. For the first time were described in details the experimental lesions produced by E. dispar, as well as the distribution of the trophozoites detected by the immunohistochemistry. Animals experimentally infected with E. dispar presented necrosis, thrombosis and chronic granulomatous inflammation. Immunoreactive products derived from trofozoites were observed close or associated with trophozoites, epithelioid cells, leucocytes and hepatocytes. Few are the articles on the literature about virulence of E. dispar, which is approximately 9 times more frequent than to E. histolytica. Variation in the virulence is, therefore expected and signalizing the need of the continuity of studies with E. dispar strains from different places in the world. Taking into account that E. dispar is a closely related species to E. histolytica, these studies could determine new elements involved with E. histolytica pathogenesis, helping us to understand better the disease.

IPSE/alpha-1 of Schistosoma mansoni egg induces enlargement of granuloma but does not alter Th2 balance after infection.

Schistosomiasis is a parasitic disease with more than 200 million people infected worldwide. The formation of granulomas around eggs trapped in the liver is the main cause of disease morbidity. Therefore, the aim of this investigation was to characterize the immunopathological response induced by the recombinant (r) IPSE/alpha-1 egg protein in mice. Herein, we have shown that splenocytes from mice immunized with rIPSE/alpha-1 produced IFN-gamma, TNF-alpha, IL-4, IL-5 and IL-13 characterizing a mixed Th1/Th2 type of immune response. Pathological analysis of the liver revealed that there was no alteration in the number of eggs and granulomas; however, we observed an increase in granuloma area in immunized mice. Furthermore, eosinophil peroxidase assay showed that there was no alteration in the eosinophil infiltration in the liver; however, n-acetyl-beta-glucosaminidase measurement revealed an increase in macrophage activity. Despite the alteration in the profile of liver inflammatory cells in rIPSE immunized mice, the production of chemokines such as CCL2, CCL3, CCL5 and CCL11 was unaltered compared with the control group. In conclusion, IPSE/alpha-1 immunization induces a mixed Th1/Th2 type of immune response and enlargement of hepatic granuloma caused by an increased macrophage activity, but does not alter Th2 cytokines following infection.

Immunization with SmIg, a novel tegument protein from Schistosoma mansoni, fails to induce protection in mice but reduces liver pathology.

Proteins associated with the schistosome tegument are of great importance for the development of new intervention strategies since they may be exposed on the surface of the parasite. Herein, we have isolated a cDNA clone encoding for the Schistosoma mansoni SmIg and its recombinant protein was tested as a potential vaccine candidate. Initially, its amino acid sequence was analysed by bioinformatics and shown to possess an N-terminal signal peptide, a C-terminal transmembrane helix, 4 glycosylation sites, an immunoglobulin conserved domain and 73% similarity with a hypothetical S. japonicum protein of unknown function. SmIg was produced by E. coli as a recombinant protein (rSmIg) and its protective effectiveness was evaluated against S. mansoni infection with 100 cercariae in a murine model. Mice immunized with rSmIg induced an immune response characterized by dominant IgG1 isotype and significant levels of IFN-gamma, TNF-alpha, IL-10 and IL-4. Although immunogenic, the recombinant vaccine failed to induce worm burden reduction when compared to the infected control group. However, rSmIg-immunized mice had significant reductions of liver granuloma volume and fibrosis content by 31.8% and 49%, respectively. In conclusion, SmIg is a new tegument protein from S. mansoni that plays an important role in reducing pathology induced by parasite infection.

Glycogen content is affected differently in acute pulmonary and extra-pulmonary lung injury.

Acute respiratory distress syndrome (ARDS) is the most severe form of acute lung injury (ALI). The aim of the present study was to investigate whether paraquat-induced acute pulmonary and extra-pulmonary lung injury (ALI-P and ALI-EX, respectively), in rats, affects glycogen content in different tissues. This measurement could indicate performance limitations of tissues, a new biochemical aspect of ARDS. ALI-P and ALI-EX were induced by injection into the trachea (0.5 mg/kg) and intraperitoneally (20 mg/kg) 24 hours prior to tissue collection. The control groups (CTRL) received the same volume of saline. Glycogen content (mg/g tissue) from different tissues was measured using the anthrone reagent. Glycogen content in the heart and kidney was higher in the ALI-EX group than the CTRL-EX group. Glycogen content in the gastrocnemius muscle was lower in the ALI-EX group than the CTRL-EX group. However, there were no significant differences in glycogen content in the diaphragm in the ALI-EX and ALI-P groups or in the gastrocnemius, heart and kidney in the ALI-P group when compared to the respective controls. ALI-EX caused a greater thickening of the alveolar walls, more areas of atelectasis and a greater abundance of inflammatory cells in comparison to ALI-P. These results demonstrate that glycogen content in ALI, induced by an herbicide that is highly toxic to humans and animals, is altered in different tissues depending on the location of the injury.

An in vivo experimental model to assess furcal lesions as a result of perforation.

AIM: To design and validate a rat molar model of furcal perforation to allow investigation of the biological phenomena that follow and to explore its potential for evaluating repair materials under standardized conditions. METHODOLOGY: Eighteen male Wistar rats were used. Surgical aseptic procedures were carried out in order to open the pulp chamber of a first molar tooth. A cavity was prepared on the floor of the pulp chamber using a (1/4) round bur that created a communication between the furcation and the periodontal tissues. Six animals for each time point were sacrificed on days 14, 21 and 28 to assay morphological changes at the furcation region of molars. Maxillary bone was processed, removed and sectioned. Cellular infiltration, collagen deposition and bone resorption were assessed by histological analysis. Cellularity in the lesion area was determined by morphometric analysis. Data were analysed using parametric Student's t-test. RESULTS: A furcal perforation model was standardized in which both radiological outcome and periodontal tissue reactions could be assessed through evaluation of cellularity, osteoclast activity and collagen deposition. The morphometric analysis revealed a greater number of cells 21 day post-surgery when compared with 14 days. CONCLUSION: This animal model was suitable for radiological and histological evaluation of the processes that accompany surgical furcal perforation.