Succinyl-CoA Synthetase: New Antigen Candidate of Bartonella bacilliformis

BACKGROUND: Bartonella bacilliformis is the causative agent of Carrion's disease, a neglected illness with mortality rates of 40-85% in the absence of treatment. The lack of a diagnostic technique to overcome misdiagnosis and treat asymptomatic carriers is of note. This study aimed to identify new B. bacilliformis antigenic candidates that could lead to a new diagnostic tool able to be implemented in endemic rural areas. METHODOLOGY/PRINCIPAL FINDINGS: Blood (n = 198) and serum (n = 177) samples were collected in northern Peru. Clinical data were recorded. Specific 16S rRNA amplification by RT-PCR, IFA and ELISA for IgM/IgG with whole cells as antigens was done. Western blot analysis and N-terminal amino acid sequencing detected seroreactive proteins. ELISAs for IgM/IgG for the antigenic candidates were performed. Of the population 33.3% reported at least one symptom compatible with Carrion's disease; 25.4% (IFA), 27.1% (ELISA-IgG), 33.9% (ELISA-IgM) and 38.9% (RT-PCR) of samples were positive. Four proteins were considered potential antigenic candidates, including two new antigenic candidates, succinyl-CoA synthetase subunit alpha (SCS-alpha) and succinyl-CoA synthetase subunit beta (SCS-beta). On Western blot both Pap31 and SCS-alpha interacted with IgM, while GroEL and SCS-beta interacted with IgG. The presence of specific antibodies against the antigenic candidates varied from 34.5% (IgG against SCS-alpha) to 97.2% (IgM against Pap31). CONCLUSIONS/SIGNIFICANCE: RT-PCR and the high levels of positivity for specific ELISAs demonstrate high levels of B. bacilliformis exposure and asymptomatic carriers among inhabitants. The new antigens identified might be used as a new rapid diagnostic tool to diagnose acute Carrion's disease and identify asymptomatic carriers

Construction of an All-in-one Double-conditional shRNA Expression Vector

Gene silencing by RNA interference （RNAi） is widely used for assessing gene function. An important advance in the RNAi field was the discovery that plasmid-based RNAi can substitute for synthetic small interfering RNA in vitro and in vivo. However, constitutive and ubiquitous knockdown of gene expression by RNAi in mice can limit the scope of experiments because this process can lead to embryonic lethality, or result in compensatory overexpression of other genes such that no phenotypic abnormalities occur. Either way, analyses of the physiological roles of the gene of interest in adult mice are not possible. To overcome these limitations, we previously constructed a double-conditional short-hairpin RNA （shRNA） expression vector that can regulate shRNA expression in a spatio-temporal manner with a tetracycline-inducible floxed stuffer sequence selectively excised by application of Cre recombinase. In this study, we aimed to modify this vector to create an all-in-one vector that produces double-conditional transgenic mice through a single round of gene transfer to fertilized eggs. We added a coding region for nuclear localizing Cre （NCre） recombinase with a multi-cloning site for a cell-specific promoter into the double-conditional short-hairpin RNA （shRNA） expression vector that we previously constructed. Using Escherichia coli, we confirmed successful construction of the vector. First, we confirmed isopropyl-β-D-thiogalactopyranoside-induced expression of NCre recombinase through the lac operon as a specific promoter by western blotting. Second, we confirmed functional recombination of the floxed sequence of loxP-like TATA-lox by analysing restriction enzyme-digested fragments. This all-in-one double-conditional shRNA expression vector will be useful for reversible in vitro and in vivo knockdown of target gene expression, in target cells via promoter-specific expression of NCre, and at specific times by tetracycline application

Small classes vs. large classes : the influence of class size on learner strategy use in an EFL context

This study explored how small English classes facilitate foreign language learning, especially speaking skills. It is generally believed that smaller classes are more appropriate for teaching/learning speaking skills. This study investigated the effects of class size using a questionnaire with 76 college students majoring in English in Japan and classroom observation. The participants were divided into two groups: those who were learning English in small classes (about 10 students per class) and a large class (about 20 students per class). The questionnaire was developed for this study based on the Strategy Inventory for Language Learning (henceforth, SILL: Oxford, 1990). As a result of this study, significant differences were found between the two types of classes in cognitive strategy use. However, use of metacognitive strategies was low in both groups. The results suggested that small classes are not a panacea for teaching/learning speaking skills, but the judicious use of teaching/learner strategies in both settings is important

Soil biochemistry and microbial activity in vineyards under conventional and organic management at Northeast Brazil.

The São Francisco Submedium Valley is located at the Brazilian semiarid region and is an important center for irrigated fruit growing. This region is responsible for 97% of the national exportation of table grapes, including seedless grapes. Based on the fact that orgThe São Francisco Submedium Valley is located at the Brazilian semiarid region and is an important center for irrigated fruit growing. This region is responsible for 97% of the national exportation of table grapes, including seedless grapes. Based on the fact that organic fertilization can improve soil quality, we compared the effects of conventional and organic soil management on microbial activity and mycorrhization of seedless grape crops. We measured glomerospores number, most probable number (MPN) of propagules, richness of arbuscular mycorrhizal fungi (AMF) species, AMF root colonization, EE-BRSP production, carbon microbial biomass (C-MB), microbial respiration, fluorescein diacetate hydrolytic activity (FDA) and metabolic coefficient (qCO2). The organic management led to an increase in all variables with the exception of EE-BRSP and qCO2. Mycorrhizal colonization increased from 4.7% in conventional crops to 15.9% in organic crops. Spore number ranged from 4.1 to 12.4 per 50 g-1 soil in both management systems. The most probable number of AMF propagules increased from 79 cm-3 soil in the conventional system to 110 cm-3 soil in the organic system. Microbial carbon, CO2 emission, and FDA activity were increased by 100 to 200% in the organic crop. Thirteen species of AMF were identified, the majority in the organic cultivation system. Acaulospora excavata, Entrophospora infrequens, Glomus sp.3 and Scutellospora sp. were found only in the organically managed crop. S. gregaria was found only in the conventional crop. Organically managed vineyards increased mycorrhization and general soil microbial activity

Ebola Zaire Virus Blocks Type I Interferon Production by Exploiting the Host SUMO Modification Machinery

Ebola Zaire virus is highly pathogenic for humans, with case fatality rates approaching 90% in large outbreaks in Africa. The virus replicates in macrophages and dendritic cells (DCs), suppressing production of type I interferons (IFNs) while inducing the release of large quantities of proinflammatory cytokines. Although the viral VP35 protein has been shown to inhibit IFN responses, the mechanism by which it blocks IFN production has not been fully elucidated. We expressed VP35 from a mouse-adapted variant of Ebola Zaire virus in murine DCs by retroviral gene transfer, and tested for IFN transcription upon Newcastle Disease virus (NDV) infection and toll-like receptor signaling. We found that VP35 inhibited IFN transcription in DCs following these stimuli by disabling the activity of IRF7, a transcription factor required for IFN transcription. By yeast two-hybrid screens and coimmunoprecipitation assays, we found that VP35 interacted with IRF7, Ubc9 and PIAS1. The latter two are the host SUMO E2 enzyme and E3 ligase, respectively. VP35, while not itself a SUMO ligase, increased PIAS1-mediated SUMOylation of IRF7, and repressed Ifn transcription. In contrast, VP35 did not interfere with the activation of NF-κB, which is required for induction of many proinflammatory cytokines. Our findings indicate that Ebola Zaire virus exploits the cellular SUMOylation machinery for its advantage and help to explain how the virus overcomes host innate defenses, causing rapidly overwhelming infection to produce a syndrome resembling fulminant septic shock

Research Activities in the Department of Nursing

Research activity at the Department of Nursing is overviewed from the point of research topics, the theme of the projects admitted for grant from the Ministry of Education and Science of Japan, and expected research topics, trying to clarify the needs and challenges of the Department from multilateral aspects in future research activities. The Department of Nursing, Aino University is currently divided into the five areas and further into 12 fields. On the other hand, according to the Scientific Research Grant Program (2015 fiscal year), the research topics in nursing science is subdivided into the five areas; a) basic nursing, b) clinical nursing, c) lifelong developmental nursing, d) elderly nursing, and e) community health nursing

早期体験学習としての基礎看護学実習の学習効果と実習満足度に関連する要因

Fundamental nursing practice, the first step of clinical practice, is important for nursing students to facilitate their early, gradual introduction to clinical practice. As part of this fundamental nursing practice, first-year students have been exposed to nursing practice on one day in June for the past 4 years. Factors related to the effects of this early exposure of students to fundamental nursing practice on their learning and levels of satisfaction with this style of clinical practice were investigated based on their self-evaluation. 1) Students\u27 relevant experiences before entering the college included admission to a hospital (31.6%) and one-day nursing experience (48.8%). 2) Nursing roles they had successfully observed included monitoring/recording/reporting, understanding the patient, daily life support, ensuring security and safety, environmental adjustment, respecting the patient\u27s independence, empathizing with the patient, assistance during treatment, mental support, cooperating with other professions, and guiding the patient. 3) The achievement levels of the practice goals "to observe the site of nursing practice" (p<0.01) and "to observe nurses actively performing duties" (p<0.05) were significantly correlated with students\u27 satisfaction levels. 4) The effectiveness of prior study or individual guidance on self-defined targets, an orientation held in the ward, and observation of nurses in service had significantly affected students\u27 satisfaction levels. A student\u27s success in understanding a nurse\u27s roles in the practice was significantly correlated with his/her satisfaction level. 5) An effective commitment of the tutor nurse to students\u27 learning had a significant impact on their satisfaction levels. 6) Students\u27 attitudes toward the clinical practice, such as viewing it as preferable, interesting, or wanting to be a nurse, were significantly correlated with their satisfaction levels. 7) Providing students with an opportunity to communicate with patients in the practice was significantly correlated with their satisfaction levels. These findings suggested that students viewed their early exposure to nursing practice as a valuable learning experience, and used it to develop themselves and prepare for the future. Since their interest in the clinical practice promotes their willingness to learn nursing, it is necessary to help them feel positive about the practice. Besides the importance of cooperation between clinical workers including the tutor nurse and teaching staff, and their explanations to students about the practice goals and levels, there is a need for those involved in teaching to review the practice contents. The necessity of appropriately designing the contents to facilitate students\u27 communication with patients was also suggested

Utilização dos princípios de adesão à dentina na obturação do sistema de canais radiculares

O objetivo deste trabalho foi revisar a literatura quanto à utilização de sistemas adesivos para a obturação do sistema de canais radiculares em tratamentos endodônticos. Atualmente, princípios da Dentística Restauradora vêm sendo avaliados para serem empregados na obturação dos canais radiculares. Materiais adesivos desenvolvidos para este fim estão sendo desenvolvidos para que possam ser utilizados com maior freqüência e segurança na prática clínica, na tentativa de superar as limitações apresentadas pelo material obturador convencionais: guta percha associada ao cimento endodôntico. Foi concluído nesta revisão de literatura que a guta percha, associada aos cimentos endodônticos convencionais, ainda pode ser considerada o material de escolha para a obturação dos canais radiculares, uma vez que nenhum sistema que prevê adesão química no interior dos canais radiculares tem apresentado resultados superiores. Novas pesquisas precisam ser realizadas para que as limitações dos novos sistemas sejam contornadas e um material obturador ideal possa ser desenvolvido.The main goal of this paper was to review the literature on the use of adhesive systems to fill the root canal systems in Endodontics. Nowadays, the Restorative Dentistry basis has been assessed to be applied in the Endodontics filling procedures. Adhesive materials have been developed to be used more frequently and safely in clinical procedures in order to go beyond the limitations of the current filling materials used by the majority of clinicians: gutta-percha with endodontic sealers. It was concluded in the present review that the gutta-percha, used with the conventional endodontic sealers, can be considered the choice material to be used in Endodontics, because none of the new materials based on adhesive principles has been shown better clinical performance. New researches must be carried on to overcome the new products limitations, searching an ideal material