Effect of Eugenol against Streptococcus agalactiae

Streptococcus agalactiae (group B streptococci (GBS)) is an important infections agent in newborns associated with maternal vaginal colonization. Intrapartum antibiotic prophylaxis in GBS-colonized pregnant women has led to a significant reduction in the incidence of early neonatal infection in various geographic regions. However, this strategy may lead to resistance selecting among GBS, indicating the need for new alternatives to prevent bacterial transmission and even to treat GBS infections. This study reported for the first time the effect of eugenol on GBS isolated from colonized women, alone and in combination with silver nanoparticles produced by Fusarium oxysporum (AgNPbio). Eugenol showed a bactericidal effect against planktonic cells of all GBS strains, and this effect appeared to be time-dependent as judged by the time-kill curves and viability analysis. Combination of eugenol with AgNPbio resulted in a strong synergistic activity, significantly reducing the minimum inhibitory concentration values of both compounds. Scanning and transmission electron microscopy revealed fragmented cells and changes in bacterial morphology after incubation with eugenol. In addition, eugenol inhibited the viability of sessile cells during biofilm formation and in mature biofilms. These results indicate the potential of eugenol as an alternative for controlling GBS infections

BRCA1 protein dose-dependent risk for embryonic oxidative DNA damage, embryopathies and neurodevelopmental disorders with and without ethanol exposure

Although widely known as a tumor suppressor, the breast cancer 1 susceptibility protein (BRCA1) is also important in development, where it regulates fetal DNA repair pathways that protect against DNA damage caused by physiological and drug-enhanced levels of reactive oxygen species (ROS). We previously showed that conditional heterozygous (+/-) knockout (cKO) mouse embryos with a minor 28% BRCA1 deficiency developed normally in culture, but when exposed to the ROS-initiating drug, alcohol (ethanol, EtOH), exhibited embryopathies not evident in wild-type (+/+) littermates. Herein, we characterized a direct Brca1 +/- knockout (KO) model with a 2-fold greater (58%) reduction in BRCA1 protein vs. the cKO model. We also characterized and compared learning & memory deficits in both the cKO and KO models. Even saline-exposed Brca1 +/- vs. +/+ KO progeny exhibited enhanced oxidative DNA damage and embryopathies in embryo culture and learning & memory deficits in females in vivo, which were not observed in the cKO model, revealing the potential pathogenicity of physiological ROS levels. The embryopathic EtOH concentration for cultured direct KO embryos was half that for cKO embryos, and EtOH affected Brca1 +/+ embryos only in the direct KO model. The spectrum and severity of EtOH embryopathies in culture were greater in both Brca1 +/- vs. +/+ embryos, and direct KO vs. cKO +/- embryos. Motor coordination deficits were evident in both male and female Brca1 +/- KO progeny exposed in utero to EtOH. The results in our direct KO model with a greater BRCA1 deficiency vs. cKO mice provide the first evidence for BRCA1 protein dose-dependent susceptibility to developmental disorders caused by physiological and drug-enhanced oxidative stress

Protein Mimetic and Anticancer Properties of Monocyte-Targeting Peptide Amphiphile Micelles

Monocyte chemoattractant protein-1 (MCP-1) stimulates the migration of monocytes to inflammatory sites, leading to the progression of many diseases. Recently, we described a monocyte-targeting peptide amphiphile micelle (MCP-1 PAM) incorporated with the chemokine receptor CCR2 binding motif of MCP-1, which has a high affinity for monocytes in atherosclerotic plaques. We further report here the biomimetic components of MCP-1 PAMs and the influence of the nanoparticle upon binding to monocytes. We report that MCP-1 PAMs have enhanced secondary structure compared to the MCP-1 peptide. As a result, MCP-1 PAMs displayed improved binding and chemoattractant properties to monocytes, which upregulated the inflammatory signaling pathways responsible for monocyte migration. Interestingly, when MCP-1 PAMs were incubated in the presence of prostate cancer cells in vitro, the particle displayed anticancer efficacy by reducing CCR2 expression. Given that monocytes play an important role in tumor cell migration and invasion, our results demonstrate that PAMs can improve the native biofunctional properties of the peptide and may be used as an effective inhibitor to prevent chemokine–receptor interactions that promote disease progression

Molecular Characterization of Trypanosoma cruzi Tc8.2 Gene Indicates Two Differential Locations for the Encoded Protein in Epimastigote and trypomastigote Forms

Submitted by Luciane Willcox ([email protected]) on 2016-08-30T15:47:05Z No. of bitstreams: 1 Molecular characterization of Trypanosoma cruzi Tc8.2 gene.pdf: 630488 bytes, checksum: 8888e97da01447ee45235bf2ed02d7b2 (MD5)Approved for entry into archive by Luciane Willcox ([email protected]) on 2016-08-30T17:21:12Z (GMT) No. of bitstreams: 1 Molecular characterization of Trypanosoma cruzi Tc8.2 gene.pdf: 630488 bytes, checksum: 8888e97da01447ee45235bf2ed02d7b2 (MD5)Made available in DSpace on 2016-08-30T17:21:12Z (GMT). No. of bitstreams: 1 Molecular characterization of Trypanosoma cruzi Tc8.2 gene.pdf: 630488 bytes, checksum: 8888e97da01447ee45235bf2ed02d7b2 (MD5) Previous issue date: 2015-08-25Universidade Estadual de Londrina. Centro de Ciências Biológicas. Departamento de Microbiologia. Londrina, PR, Brasil.Universidade Estadual de Londrina. Centro de Ciências Biológicas. Departamento de Microbiologia. Londrina, PR, Brasil.Universidade Estadual de Londrina. Centro de Ciências Biológicas. Departamento de Microbiologia. Londrina, PR, Brasil.Universidade Federal de São Paulo. Departamento de Microbiologia, Imunobiologia e Parasitologia. Londrina, PR, Brasil.Universidade Estadual de Londrina. Centro de Ciências Biológicas. Departamento de Ciências Patológicas. Laboratório de Imunologia. Londrina, PR, Brasil / Fundação Oswaldo Cruz. Instituto Carlos Chagas. Curitiba, PR, Brasil.Universidade Bandeirante de São Paulo. Conselho de Pós-graduação e Pesquisa. São Paulo, SP, Brasil.Universidade Federal de São Paulo. Departamento de Microbiologia, Imunobiologia e Parasitologia. Londrina, PR, Brasil.Universidade Estadual de Maringá. Centro de Ciências da Saúde. Departamento de Ciências Básicas da Saúde. Maringá, PR, Brasil.Universidade de São Paulo. Departamento de Parasitologia, Microbiologia e Imunologia. Ribeirão Preto, SP, Brasil.Universidade Estadual de Londrina. Centro de Ciências Biológicas. Departamento de Microbiologia. Londrina, PR, Brasil.Universidade Estadual de Londrina. Centro de Ciências Biológicas. Departamento de Microbiologia. Londrina, PR, Brasil.This report describes the molecular characterization of the Tc8.2 gene of Trypanosoma cruzi. Both the Tc8.2 gene and its encoded protein were analyzed by bioinformatics, while Northern blot and RT-PCR were used for the transcripts. Besides, immunolocalization of recombinant protein was done by immunofluorescence and electron microscopy. Analysis indicated the presence of a single copy of Tc8.2 in the T. cruzi genome and 2-different sized transcripts in epimastigotes/amastigotes and trypomastigotes. Immunoblotting showed 70 and 80 kDa polypeptides in epimastigotes and trypomastigotes, respectively, and a differential pattern of immunolocalization. Overall, the results suggest that Tc8.2 is differentially expressed during the T. cruzi life cycle

Effect of eugenol against streptococcus agalactiae and synergistic interaction with biologically produced silver nanoparticles

Streptococcus agalactiae (group B streptococci (GBS)) is an important infections agent in newborns associated with maternal vaginal colonization. Intrapartumantibiotic prophylaxis in GBS-colonized pregnant women has led to a significant reduction in the incidence of early neonatal infection in various geographic regions. However, this strategy may lead to resistance selecting among GBS, indicating the need for new alternatives to prevent bacterial transmission and even to treat GBS infections. This study reported for the first time the effect of eugenol on GBS isolated from colonized women, alone and in combination with silver nanoparticles produced by Fusarium oxysporum (AgNPbio). Eugenol showed a bactericidal effect against planktonic cells of all GBS strains, and this effect appeared to be time-dependent as judged by the time-kill curves and viability analysis. Combination of eugenol with AgNPbio resulted in a strong synergistic activity, significantly reducing the minimum inhibitory concentration values of both compounds. Scanning and transmission electron microscopy revealed fragmented cells and changes in bacterial morphology after incubation with eugenol. In addition, eugenol inhibited the viability of sessile cells during biofilm formation and in mature biofilms. These results indicate the potential of eugenol as an alternative for controlling GBS infections2015CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICO - CNPQCOORDENAÇÃO DE APERFEIÇOAMENTO DE PESSOAL DE NÍVEL SUPERIOR - CAPES402728/2013-0sem informaçãoPrograma de Pos-Graduacao em Microbiologia da Universidade Estadual de Londrin

Exposure to silver nanospheres leads to altered respiratory mechanics and delayed immune response in an in vivo Murine model

Here we examine the organ level toxicology of both carbon black (CB) and silver nanoparticles (AgNP). We aim to determine metal-specific effects to respiratory function, inflammation and potential interactions with lung lining fluid (LLF). C57Bl6/J male mice were intratracheally instilled with saline (control), low (0.05 μg/g) or high (0.5 μg/g) doses of either AgNP or CB 15 nm nanospheres. Lung histology, cytology, surfactant composition and function, inflammatory gene expression, and pulmonary function were measured at 1, 3, and 7 days post-exposure. Acutely, high dose CB resulted in an inflammatory response, increased neutrophilia and cytokine production, without alteration in surfactant composition or respiratory mechanics. Low dose CB had no effect. Neither low nor high dose AgNPs resulted in an acute inflammatory response, but there was an increase in work of breathing. Three days post-exposure with CB, a persistent neutrophilia was noted. High dose AgNP resulted in an elevated number of macrophages and invasion of lymphocytes. Additionally, AgNP treated mice displayed increased expression of IL1B, IL6, CCL2, and IL10. However, there were no significant changes in respiratory mechanics. At day 7, inflammation had resolved in AgNP-treated mice, but tissue stiffness and resistance were significantly decreased, which was accompanied by an increase in surfactant protein D (SP-D) content. These data demonstrate that the presence of metal alters the response of the lung to nanoparticle exposure. AgNP-surfactant interactions may alter respiratory function and result in a delayed immune response, potentially due to modified airway epithelial cell function

<i>T.</i><i>cruzi</i> infection induces thrombocytopenia and leukopenia transients in the early of infection.

<p>C5BL/6 mice were infected with 5×10³ trypomastigotes (Y strain) via i.p. injection, monitored for the development of thrombocytopenia and leukopenia, and sacrificed at different time points post-<i>T. cruzi</i> infection. <i>A</i>: platelets and <i>B</i>: leukocytes were counts from peripheral blood from uninfected and infected mice. Values represent the mean ± standard error and are representative of three independent experiments, using 4–15 mice per group. Results were analyzed by analysis of variance (ANOVA) followed by Bonferroni multiple comparisons test. Asterisks indicates significant differences (<i>p<</i>0.05) when compared with control group (uninfected).</p

TS reduces blood platelets counts in naïve C57BL/6 mice.

<p>Mice were inoculated i.p with 50 µg of recombinant TS. <i>A</i>: platelet, <i>B</i>: leukocyte and <i>C</i>: megakariocyte counts were determined 24 h later. Values represent the mean ± standard error and are representative of two independent experiments; using 7–15 mice per group Results were analyzed by analysis of variance (ANOVA) followed by Bonferroni multiple comparisons test. Asterisks indicate significant differences (<i>p</i><0.05).</p