Neutrophil recruitment in endotoxin-induced murine mastitis is strictly dependent on mammary alveolar macrophages

Mastitis, inflammation of the mammary tissue, is a common disease in dairy animals and mammary pathogenic Escherichia coli (MPEC) is a leading cause of the disease. Lipopolysaccharide (LPS) is an important virulence factor of MPEC and inoculation of the mammary glands with bacterial LPS is sufficient to induce an inflammatory response. We previously showed using adoptive transfer of normal macrophages into the mammary gland of TLR4-deficient C3H/HeJ mice that LPS/TLR4 signaling on mammary alveolar macrophages is sufficient to elicit neutrophil recruitment into the alveolar space. Here we show that TLR4-normal C3H/HeN mice, depleted of alveolar macrophages, were completely refractory to LPS intramammary challenge. These results indicate that alveolar macrophages are both sufficient and essential for neutrophil recruitment elicited by LPS/TLR4 signaling in the mammary gland. Using TNFα gene-knockout mice and adoptive transfer of wild-type macrophages, we show here that TNFα produced by mammary alveolar macrophages in response to LPS/TLR4 signaling is an essential mediator eliciting blood neutrophil recruitment into the milk spaces. Furthermore, using the IL8 receptor or IL1 receptor gene-knockout mice we observed abrogated recruitment of neutrophils into the mammary gland and their entrapment on the basal side of the alveolar epithelium in response to intramammary LPS challenge. Adoptive transfer of wild-type neutrophils to IL1 receptor knockout mice, just before LPS challenge, restored normal neutrophil recruitment into the milk spaces. We conclude that neutrophil recruitment to the milk spaces is: (i) mediated through TNFα, which is produced by alveolar macrophages in response to LPS/TLR4 signaling and (ii) is dependent on IL8 and IL1β signaling and regulated by iNOS-derived NO

Principios de enseñanza

Este folleto contiene información sobre los métodos de enseñanza más efectivos y presenta a su vez, diez principios de instrucción basados en diversas investigaciones. Estos principios provienen de tres fuentes de información distintas: (a) investigación sobre la forma en que el cerebro almacena y utiliza la información nueva; (b) investigación sobre las prácticas en el aula de aquellos docentes cuyos alumnos muestran el mayor provecho en materia de aprendizaje; y (c) resultados de estudios relacionado con las estrategias de aprendizaje de los alumnos. También se incluyen algunas sugerencias relativas a buenas prácticas en el aula. Si bien este folleto está basado en investigaciones realizadas principalmente en países económicamente más avanzados, éste se enfoca en aspectos universales del aprendizaje de la lengua y de la instrucción; por lo tanto, las prácticas sugeridas pueden ser adoptadas también en otros contextos

Immigration Federalism: A Reappraisal

This Article identifies how the current spate of state and local regulation is changing the way elected officials, scholars, courts, and the public think about the constitutional dimensions of immigration law and governmental responsibility for immigration enforcement. Reinvigorating the theoretical possibilities left open by the Supreme Court in its 1875 Chy Lung v. Freeman decision, state and local offi- cials characterize their laws as unavoidable responses to the policy problems they face when they are squeezed between the challenges of unauthorized migration and the federal government’s failure to fix a broken system. In the October 2012 term, in Arizona v. United States, the Court addressed, but did not settle, the difficult empirical, theoretical, and constitutional questions necessitated by these enactments and their attendant justifications. Our empirical investigation, however, discovered that most state and local immigration laws are not organic policy responses to pressing demographic challenges. Instead, such laws are the product of a more nuanced and politicized process in which demographic concerns are neither neces- sary nor sufficient factors and in which federal inactivity and subfederal activity are related phenomena, fomented by the same actors. This Article focuses on the con- stitutional and theoretical implications of these processes: It presents an evidence- based theory of state and local policy proliferation; it cautions legal scholars to rethink functionalist accounts for the rise of such laws; and it advises courts to reassess their use of traditional federalism frameworks to evaluate these sub federal enactments

Anatomy Teaching, a “Model” Answer? Evaluating “Geoff”, a Painted Anatomical Horse, as a Tool for Enhancing Topographical Anatomy Learning

Development of new methods for anatomy teaching is increasingly important as we look to modernize and supplement traditional teaching methods. In this study, a life‐sized equine model, “Geoff” was painted with surface and deep anatomical structures with the aim of improving students’ ability to convert theoretical knowledge into improved topographical anatomy knowledge on the live horse. Third and fourth year veterinary medicine students (n = 45) were randomly allocated into experimental (used “Geoff”) and control (used textbook) groups. The efficacy of the model was evaluated through a structured oral exam using a live horse. Questionnaires gathered information on student confidence and enjoyment of the task. There was no significant difference in the performance of experimental and control groups either immediately (44±20% vs. 40±21%; P = 0.504) or 9 weeks after the learning intervention (55±17% vs. 55±20%; P = 0.980). There were however specific questions on which the experimental group performed better than controls, and for which gender effects were apparent. The students using “Geoff” showed a transient gain in confidence following the session (Likert scale 2.7 to 3.6) however the initial increase was no longer present at the second test. There was a significant influence of gender on confidence with greater confidence gains in females in the Experimental group. The students found the model to be extremely useful and both groups found the sessions enjoyable. The model will be of benefit as a complementary learning tool for students

Structure of GrlR and the Implication of Its EDED Motif in Mediating the Regulation of Type III Secretion System in EHEC

Enterohemorrhagic Escherichia coli (EHEC) is a common cause of severe hemorrhagic colitis. EHEC's virulence is dependent upon a type III secretion system (TTSS) encoded by 41 genes. These genes are organized in several operons clustered in the locus of enterocyte effacement. Most of the locus of enterocyte effacement genes, including grlA and grlR, are positively regulated by Ler, and Ler expression is positively and negatively modulated by GrlA and GrlR, respectively. However, the molecular basis for the GrlA and GrlR activity is still elusive. We have determined the crystal structure of GrlR at 1.9 Å resolution. It consists of a typical β-barrel fold with eight β-strands containing an internal hydrophobic cavity and a plug-like loop on one side of the barrel. Strong hydrophobic interactions between the two β-barrels maintain the dimeric architecture of GrlR. Furthermore, a unique surface-exposed EDED (Glu-Asp-Glu-Asp) motif is identified to be critical for GrlA–GrlR interaction and for the repressive activity of GrlR. This study contributes a novel molecular insight into the mechanism of GrlR function

A framework to analyze argumentative knowledge construction in computer-supported collaborative learning

Computer-supported collaborative learning (CSCL) is often based on written argumentative discourse of learners, who discuss their perspectives on a problem with the goal to acquire knowledge. Lately, CSCL research focuses on the facilitation of specific processes of argumentative knowledge construction, e.g., with computer-supported collaboration scripts. In order to refine process-oriented instructional support, such as scripts, we need to measure the influence of scripts on specific processes of argumentative knowledge construction. In this article, we propose a multi-dimensional approach to analyze argumentative knowledge construction in CSCL from sampling and segmentation of the discourse corpora to the analysis of four process dimensions (participation, epistemic, argumentative, social mode)

Specifying computer-supported collaboration scripts

Collaboration scripts are activity programs which aim to foster collaborative learning by structuring interaction between learners. Computer-supported collaboration scripts generally suffer from the problem of being restrained to a specific learning platform and learning context. A standardization of collaboration scripts first requires a specification of collaboration scripts that integrates multiple perspectives from computer science, education and psychology. So far, only few and limited attempts at such specifications have been made. This paper aims to consolidate and expand these approaches in light of recent findings and to propose a generic framework for the specification of collaboration scripts. The framework enables a description of collaboration scripts using a small number of components (participants, activities, roles, resources and groups) and mechanisms (task distribution, group formation and sequencing)

EspZ of enteropathogenic and enterohemorrhagic Escherichia coli regulates type III secretion system protein translocation.

UNLABELLED: Translocation of effector proteins via a type III secretion system (T3SS) is a widespread infection strategy among Gram-negative bacterial pathogens. Each pathogen translocates a particular set of effectors that subvert cell signaling in a way that suits its particular infection cycle. However, as effector unbalance might lead to cytotoxicity, the pathogens must employ mechanisms that regulate the intracellular effector concentration. We present evidence that the effector EspZ controls T3SS effector translocation from enteropathogenic (EPEC) and enterohemorrhagic (EHEC) Escherichia coli. Consistently, an EPEC espZ mutant is highly cytotoxic. Following ectopic expression, we found that EspZ inhibited the formation of actin pedestals as it blocked the translocation of Tir, as well as other effectors, including Map and EspF. Moreover, during infection EspZ inhibited effector translocation following superinfection. Importantly, while EspZ of EHEC O157:H7 had a universal "translocation stop" activity, EspZ of EPEC inhibited effector translocation from typical EPEC strains but not from EHEC O157:H7 or its progenitor, atypical EPEC O55:H7. We found that the N and C termini of EspZ, which contains two transmembrane domains, face the cytosolic leaflet of the plasma membrane at the site of bacterial attachment, while the extracellular loop of EspZ is responsible for its strain-specific activity. These results show that EPEC and EHEC acquired a sophisticated mechanism to regulate the effector translocation. IMPORTANCE: Enteropathogenic Escherichia coli (EPEC) and enterohemorrhagic E. coli (EHEC) are important diarrheal pathogens responsible for significant morbidity and mortality in developing countries and the developed world, respectively. The virulence strategy of EPEC and EHEC revolves around a conserved type III secretion system (T3SS), which translocates bacterial proteins known as effectors directly into host cells. Previous studies have shown that when cells are infected in two waves with EPEC, the first wave inhibits effector translocation by the second wave in a T3SS-dependent manner, although the factor involved was not known. Importantly, we identified EspZ as the effector responsible for blocking protein translocation following a secondary EPEC infection. Interestingly, we found that while EspZ of EHEC can block protein translocation from both EPEC and EHEC strains, EPEC EspZ cannot block translocation from EHEC. These studies show that EPEC and EHEC employ a novel infection strategy to regulate T3SS translocation

Collaboration scripts - a conceptual analysis

This article presents a conceptual analysis of collaboration scripts used in face-to-face and computer-mediated collaborative learning. Collaboration scripts are scaffolds that aim to improve collaboration through structuring the interactive processes between two or more learning partners. Collaboration scripts consist of at least five components: (a) learning objectives, (b) type of activities, (c) sequencing, (d) role distribution, and (e) type of representation. These components serve as a basis for comparing prototypical collaboration script approaches for face-to-face vs. computer-mediated learning. As our analysis reveals, collaboration scripts for face-to-face learning often focus on supporting collaborators in engaging in activities that are specifically related to individual knowledge acquisition. Scripts for computer-mediated collaboration are typically concerned with facilitating communicative-coordinative processes that occur among group members. The two lines of research can be consolidated to facilitate the design of collaboration scripts, which both support participation and coordination, as well as induce learning activities closely related to individual knowledge acquisition and metacognition. In addition, research on collaboration scripts needs to consider the learners’ internal collaboration scripts as a further determinant of collaboration behavior. The article closes with the presentation of a conceptual framework incorporating both external and internal collaboration scripts

Teaching strategies and gender in higher education instrumental studios

This study investigates instrumental music teaching strategies in higher education settings, in order to identify those employed and their frequency and context of use. An instrument- and gender-balanced sample of 24 lessons from five institutions was analysed using a researcher-designed observational instrument. The results reveal the predominance of teacher demonstration, general directives and praise as most frequent teaching strategies employed in lessons. Gender differences emerged in the teaching approaches: the male teachers gave more general directives and explanations and the female teachers offered more answers and practice discussions; the male students received the most specific teacher criticism despite uniform use of praise. The findings provide new evidence of teaching practices in advanced instrumental studios and raise questions regarding gender issues in music teaching