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Summary A surface marker that distinctly identifies cardiac progenitors (CPs) is essential for the robust isolation of these cells, circumventing the necessity of genetic modification. Here, we demonstrate that a Glycosylphosphatidylinositol-anchor containing neurotrophic factor receptor, Glial cell line-derived neurotrophic factor receptor alpha 2 ( Gfra2 ), specifically marks CPs. GFRA2 expression facilitates the isolation of CPs by fluorescence activated cell sorting from differentiating mouse and human pluripotent stem cells. Gfra2 mutants reveal an important role for GFRA2 in cardiomyocyte differentiation and development both in vitro and in vivo. Mechanistically, the cardiac GFRA2 signaling pathway is distinct from the canonical pathway dependent on the RET tyrosine kinase and its established ligands. Collectively, our findings establish a platform for investigating the biology of CPs as a foundation for future development of CP transplantation for treating heart failure. [ABSTRACT FROM AUTHOR]

Abstract: Background: Endothelin-1 (ET-1), a potent vasoconstrictor, is considered to be implicated in failing Fontan circulation, however the expressions of ET-1 and endothelin receptor type A (ETAR) and type B (ETBR) in the pulmonary arteries of failed Fontan patients were not elucidated. Methods: Immunohistochemistry and quantitative real-time PCR were used to analyse the expression levels of ET-1 and its receptors in the pulmonary arteries of the autopsy lung tissues of the patients who died after the Fontan procedure (n=10). We divided these patients into 3 groups, failed Fontan (n=4), heart failure (n=3) and non-failed Fontan (n=3), and then compared those to the age-matched normal controls (n=4). Results: The intra-acinar pulmonary arteries of failed Fontan patients showed significant medial hypertrophy. Computational optical density analyses of the immunostaining revealed that the expressions of ET-1, ETAR, and ETBR in the intra-acinar pulmonary arteries were significantly increased in the failed Fontan patients (P<0.05 vs. normal controls), however no significant difference was observed between the non-failed Fontan patients and the normal controls. Quantitative real-time PCR analyses confirmed that the mRNA expressions of ET-1, ETAR, and ETBR were significantly increased in the failed Fontan patients (P<0.05 vs. normal controls). Conclusion: The overexpression of ET-1 and its receptors in the pulmonary arteries can cause pulmonary vasoconstriction and vascular remodelling, leading to failed Fontan circulation. This study suggests a histopathological rationale for the potential benefits of endothelin receptor antagonists in patients with failing Fontan circulation. [Copyright &y& Elsevier]

Background Elucidating public concerns regarding vaccinations is important for successful immunization programs. The objective of the present study was to categorize public concerns regarding influenza vaccinations in Japan by analyzing a massive web-based question dataset. Methods The Yahoo! Chiebukuro (Japanese Yahoo! Answers) Dataset, which includes more than 16 million questions collected between April 2004 and April 2009, was used in this study. We sequentially filtered data to obtain questions on influenza vaccinations. Any questions that met our exclusion criteria concerning veterinary vaccines or computer virus vaccines were removed from the analysis. Filtered questions and their answers were manually analyzed for their content by a team of board-certified pediatricians. Results After filtering data, we obtained 1950 questions regarding influenza vaccinations. The three most frequently asked questions were regarding the vaccination schedule, safety, and effectiveness. When we analyzed monthly trends in question contents, we noted the emergence of similar questions in the same period every year. Therefore, we classified the time periods of each year into three parts: (1) from April to the commencement of seasonal influenza vaccinations (September), (2) from October until the epidemic period, and (3) the epidemic period. Two interesting results were obtained: concerns regarding effectiveness abruptly increased during the epidemic period, and pregnant or breastfeeding women increasingly asked questions regarding feasibility between October and the epidemic period. Conclusions The questions and concerns collected and analyzed in this study illustrate that the public have questions about the influenza vaccine and also that questions changed with periodical consistency. These results highlight the possible usefulness of providing the public with the latest and correct information to their questions in a timely manner, for example via an official health website. [ABSTRACT FROM AUTHOR]

Abstract: Chagas’ disease, caused by infection with the protozoan parasite Trypanosoma cruzi (T. cruzi), is intractable showing a high mortality rate, and the development of effective vaccines is much desired. To examine the efficacy of a new mode of recombinant viral vaccine, we constructed two non-transmissible Sendai viruses (rSeV/dF) encoding the full-length parasite antigen amastigote surface protein-2 (ASP2) or ASP2 fused with a mono-ubiquitin on its N-terminus (UASP2). C57BL/6 mice immunized intranasally with rSeV/dF expressing either ASP2 or UASP2 showed significantly suppressed parasitemia and could be protected from lethal T. cruzi challenge. Depletion of CD8+ T cells around the time of infection with T. cruzi completely abolished this protection, confirming that acquired immunity against the infection of T. cruzi is dependent on CD8+ T cells. We also demonstrated that the protective immunity correlated with higher secretion of interferon-γ (IFN-γ) by spleen cells on in vitro-specific or non-specific stimulation. Increased CTL activity was also confirmed by degranulation or CTL assays. Interestingly, the control virus, rSeV/dF-GFP, induced even a higher IFN-γ production from spleen cells following non-specific but not specific stimulation in vitro, suggesting that SeV may also be a good adjuvant when used as a vaccine vehicle. Taking together, the current findings indicate that recombinant Sendai virus expressing the ASP2 or UASP2 antigens of T. cruzi are interesting candidates for the development of a new mode of recombinant viral vaccine against Chagas’ disease. [Copyright &y& Elsevier]

Abstract: In vitro and in vivo studies were conducted to evaluate the effects of thiabendazole, mebendazole, levamisole and ivermectin against Gongylonema pulchrum. For in vitro assays, third-stage larvae (L3) incubated with the drugs were administered orally to mice and the ability of larvae to invade the gastric mucosa of the animals was examined. After incubation, only those larvae treated with high concentrations of levamisole (1 and 10μg/ml) were tightly coiled with intestines exhibiting morphological abnormalities. Good dose–response data for the drugs tested was observed at the time of worm recovery from mice, with no worms recovered at the two highest concentrations of levamisole. In vivo efficacy of the drugs against adult worms was evaluated in six groups of three rabbits, each of which was infected with 30 L3 of G. pulchrum and treated with thiabendazole at 100mg/kg for 3 days, mebendazole at 70mg/kg for 3 days, levamisole as a single dose of 8mg/kg, and subcutaneously injected ivermectin as a single dose of 0.2mg/kg or vehicles of the drugs (control) at 4 months post-infection. Necropsy 14 days after treatment revealed that levamisole, mebendazole and ivermectin reduced worm burdens by 63.2%, 22.8% and 25.8%, respectively, with no reductions in worms observed with thiabendazole. The surviving worms were principally found in the esophagus with the remainder distributed among the buccal mucosa, the tongue, and/or pharyngeal mucosa in all groups. A number of morphologically abnormal eggs were observed within the uterus and ovijector in female worms recovered from the thiabendazole-treated group. These findings suggest that levamisole exhibits in vivo efficacy against G. pulchrum infection and that the larval invasion tests using mice could be used to screen for anthelmintic susceptibility of nematodes. [Copyright &y& Elsevier]

On the basis of a hypothesis that proteins encoded by the mRNAs that are transported to and translated at the dendrites/synapses may play key roles in synaptic plasticity, this study reports on attempts to isolate mRNAs which are localizing at the dendrites/synapses from mouse cerebellar synaptosomal fractions. Among 100 pieces of dendritic mRNA candidates, 10 pieces of mRNAs were found to contain the cytoplasmic polyadenylation element (CPE)-like sequences which were contained in certain mRNAs translated in dendrites. We next examined the issue of whether the CPE-like sequence-containing mRNAs (CPERs) were localized in the synapses/dendrites by means of in situ hybridization. The findings indicate that CPER9 was actually localized at the apical dendrites of a portion of cerebral cortex layer V pyramidal cells, as well as at the proximal dendrites of some of the cerebellar Purkinje cells. CPER9 was found to encode a mouse homolog of MRG15, a nuclear protein which contains a chromodomain identified in several proteins that act as regulators of transcription. Immunohistochemistry with anti-MRG15 antibodies revealed that MRG15 was localized in dendrites as well as in the nuclei of Purkinje cells. These results suggest that MRG15 may serve as a link between synaptic activity and gene expression. [Copyright &y& Elsevier]