TLR induces reorganization of the IgM-BCR complex regulating murine B-1 cell responses to infections.

In mice, neonatally-developing, self-reactive B-1 cells generate steady levels of natural antibodies throughout life. B-1 cells can, however, also rapidly respond to infections with increased local antibody production. The mechanisms regulating these two seemingly very distinct functions are poorly understood, but have been linked to expression of CD5, an inhibitor of BCR-signaling. Here we demonstrate that TLR-mediated activation of CD5+ B-1 cells induced the rapid reorganization of the IgM-BCR complex, leading to the eventual loss of CD5 expression, and a concomitant increase in BCR-downstream signaling, both in vitro and in vivo after infections of mice with influenza virus and Salmonella typhimurium. Both, initial CD5 expression and TLR-mediated stimulation, were required for the differentiation of B-1 cells to IgM-producing plasmablasts after infections. Thus, TLR-mediated signals support participation of B-1 cells in immune defense via BCR-complex reorganization

Methods for Detecting Microbial Methane Production and Consumption by Gas Chromatography

Methane is an energy-dense fuel but is also a greenhouse gas 25 times more detrimental to the environment than CO2. Methane can be produced abiotically by serpentinization, chemically by Sabatier or Fisher-Tropsh chemistry, or biotically by microbes (Berndt et al., 1996; Horita and Berndt, 1999; Dry, 2002; Wolfe, 1982; Thauer, 1998; Metcalf et al., 2002). Methanogens are anaerobic archaea that grow by producing methane gas as a metabolic byproduct (Wolfe, 1982; Thauer, 1998). Our lab has developed and optimized three different gas chromatograph-utilizing assays to characterize methanogen metabolism (Catlett et al., 2015). Here we describe the end point and kinetic assays that can be used to measure methane production by methanogens or methane consumption by methanotrophic microbes. The protocols can be used for measuring methane production or consumption by microbial pure cultures or by enrichment cultures

Glycogen synthase kinase-3 regulation of chromatin segregation and cytokinesis in mouse preimplantation embryos

Glycogen synthase kinase-3 (GSK-3) is a highly conserved serine/threonine protein kinase implicated in diverse cellular processes. Activity of GSK-3 is essential for meiotic chromatin segregation in oocytes, yet expression and/or function of GSK-3 have not been reported in mammalian preimplantation embryos. Objectives of this study were to characterize GSK-3 protein expression/phosphorylation in mouse preimplantation embryos, to assess the effect of GSK-3 activity inhibition on early mitotic events, and to differentiate nuclear and cytoplasmic anomalies in GSK-3 inhibited embryos. Both GSK-3 isoforms were expressed during embryo development, with a differential expression of Α versus Β. Phosphorylation of GSK-3Α/Β at residues Y279/Y216 indicated constitutive activation throughout preimplantation development. Phosphorylation at N-terminal residues S21/S9 indicated inhibition of GSK-3Α/Β activity that was differentially regulated during early development; both Α and Β isoforms were phosphorylated during early divisions, whereas at the blastocyst stage, only Β was phosphorylated. Cytoplasmic microinjection of zygotes with anti-GSK-3Α/Β antibody significantly compromised embryonic development past the two-cell stage compared to controls. Reversibility of developmental block was tested via pharmacological inhibitors of GSK-3, lithium chloride (LiCl) and alsterpaullone. Similar to immunoneutralization, significantly fewer zygotes cultured with either LiCl or alsterpaullone developed past the two-cell stage compared to controls and this mitotic block was not reversible. Inhibition of GSK-3 activity significantly compromised timing of pronuclear membrane breakdown and mitosis initiation, nuclear development, and cytokinesis. Inhibition of GSK-3 also resulted in abnormal chromatin segregation, evidenced by incomplete karyokinesis and micronuclei formation. These results suggest that GSK-3 activity is critical for early preimplantation embryonic development. Mol. Reprod. Dev. © 2006 Wiley-Liss, Inc.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/55901/1/20495_ftp.pd

Methods for Detecting Microbial Methane Production and Consumption by Gas Chromatography

Methane is an energy-dense fuel but is also a greenhouse gas 25 times more detrimental to the environment than CO2. Methane can be produced abiotically by serpentinization, chemically by Sabatier or Fisher-Tropsh chemistry, or biotically by microbes (Berndt et al., 1996; Horita and Berndt, 1999; Dry, 2002; Wolfe, 1982; Thauer, 1998; Metcalf et al., 2002). Methanogens are anaerobic archaea that grow by producing methane gas as a metabolic byproduct (Wolfe, 1982; Thauer, 1998). Our lab has developed and optimized three different gas chromatograph-utilizing assays to characterize methanogen metabolism (Catlett et al., 2015). Here we describe the end point and kinetic assays that can be used to measure methane production by methanogens or methane consumption by methanotrophic microbes. The protocols can be used for measuring methane production or consumption by microbial pure cultures or by enrichment cultures

Universal Data System to Increase Staff Buy-In

Although PBIS has been implemented in many schools across the state, there is still a concern of making the system universal to help increase staff buy-in. Therefore, it is crucial that schools include the entire staff with the behavior expectations for all students in the school. It is also important for students to be rewarded for putting forth their best effort. Research shows that there are several products that can be purchased to allow a school to collect the data, reward points or implement incentives. However, there are cost efficient ways of implementing a school-wide data system. One strategy is to utilize the Google Suite for educators. Applications such as Forms and Sheets can be linked to provide instant access to data in real time for students and staff. Data presentations via Slides allow school teachers and administrators to analyze data to better serve their students. Teacher driven data collection also supports PBIS Tier I, Tier II and Tier III processes when implemented with fidelity. In our proposal presentation, we will discuss the importance of having a universal data system to evaluate the efficacy of PBIS practices. We will address how Google forms, Sheets, and Slides can be utilized to create a school-wide data system and can foster buy-in from staff, which can benefit the entire climate and culture of the school

Teacher Perceptions of the Instructional Leadership Practices of Principals

In today’s world school leadership, particularly instructional leadership, has taken on a new look. The era of high- stakes accountability has changed almost everything. The instructional leader of the 80’s was presented as an efficient top-down, task oriented manager who was focused on curriculum and instruction rather than buildings and budgets (Lashway, 2002). Gone are the days when principals spent most of their time with bus schedules, fire drills, and general curriculum, says the National Association of Elementary Principals (Henry, 2001). Leaders must keep abreast of state and federal goals, the latest technologies and teaching practices, as well as learn to use data to identify learning gaps among all students

Functional equivalence of stem cell and stem cell-derived extracellular vesicle transplantation to repair the irradiated brain.

Cranial radiotherapy, although beneficial for the treatment of brain tumors, inevitably leads to normal tissue damage that can induce unintended neurocognitive complications that are progressive and debilitating. Ionizing radiation exposure has also been shown to compromise the structural integrity of mature neurons throughout the brain, an effect believed to be at least in part responsible for the deterioration of cognitive health. Past work has shown that cranially transplanted human neural stem cells (hNSCs) or their extracellular vesicles (EVs) afforded long-term beneficial effects on many of these cognitive decrements. To provide additional insight into the potential neuroprotective mechanisms of cell-based regenerative strategies, we have analyzed hippocampal neurons for changes in structural integrity and synaptic remodeling after unilateral and bilateral transplantation of hNSCs or EVs derived from those same cells. Interestingly, hNSCs and EVs similarly afforded protection to host neurons, ameliorating the impact of irradiation on dendritic complexity and spine density for neurons present in both the ipsilateral and contralateral hippocampi 1 month following irradiation and transplantation. These morphometric improvements were accompanied by increased levels of glial cell-derived growth factor and significant attenuation of radiation-induced increases in postsynaptic density protein 95 and activated microglia were found ipsi- and contra-lateral to the transplantation sites of the irradiated hippocampus treated with hNSCs or hNSC-derived EVs. These findings document potent far-reaching neuroprotective effects mediated by grafted stem cells or EVs adjacent and distal to the site of transplantation and support their potential as therapeutic agents to counteract the adverse effects of cranial irradiation

Preparing for and Responding to Disturbance: Examples from the Forest Sector in Sweden and Canada

Coping or adaptation following large-scale disturbance may depend on the political system and its preparedness and policy development in relation to risks. Adaptive or foresight planning is necessary in order to account and plan for potential risks that may increase or take place concurrently with climate change. Forests constitute relevant examples of large-scale renewable resource systems that have been directly affected by recent environmental and social changes, and where different levels of management may influence each other. This article views disturbances in the forest sectors of Sweden and Canada, two large forest nations with comparable forestry experiences, in order to elucidate the preparedness and existing responses to multiple potential stresses. The article concludes that the two countries are exposed to stresses that indicate the importance of the governing and institutional system particularly with regard to multi-level systems including federal and EU levels. While economic change largely results in privatization of risk onto individual companies and their economic resources (in Canada coupled with a contestation of institutional systems and equity in these), storm and pest outbreaks in particular challenge institutional capacities at administrative levels, within the context provided by governance and tenure systems.In Sweden, funding from the research agency FORMAS, the MISTRA Arctic Futures programme, and the Future Forests programme (funded by research agency MISTRA, the forest industries, Umeå University and the Swedish University of Agricultural Sciences) as well as from the EU for data collection is acknowledged. Ryan Bullock acknowledges funding provided by the Social Sciences and Humanities Research Council and by the Mistra Arctic Futures programme for preparing this paper.https://www.mdpi.com/1999-4907/2/2/50

Distinctive expression patterns of 185/333 genes in the purple sea urchin, Strongylocentrotus purpuratus: an unexpectedly diverse family of transcripts in response to LPS, β-1,3-glucan, and dsRNA

BACKGROUND: A diverse set of transcripts called 185/333 is strongly expressed in sea urchins responding to immune challenge. Optimal alignments of full-length 185/333 cDNAs requires the insertion of large gaps that define 25 blocks of sequence called elements. The presence or absence of individual elements also defines a specific element pattern for each message. Individual sea urchins were challenged with pathogen associated molecular patterns (PAMPs) (lipopolysaccharide, β-1,3-glucan, or double stranded RNA), and changes in the 185/333 message repertoire were followed over time. RESULTS: Each animal expressed a diverse set of 185/333 messages prior to challenge and a 0.96 kb message was the predominant size after challenge. Sequence analysis of the cloned messages indicated that the major element pattern expressed in immunoquiescent sea urchins was either C1 or E2.1. In contrast, most animals responding to lipopolysaccharide, β-1,3-glucan or injury, predominantly expressed messages of the E2 pattern. In addition to the major patterns, extensive element pattern diversity was observed among the different animals before and after challenge. Nucleotide sequence diversity of the transcripts increased in response to β-1,3-glucan, double stranded RNA and injury, whereas diversity decreased in response to LPS. CONCLUSION: These results illustrate that sea urchins appear to be able to differentiate among different PAMPs by inducing the transcription of different sets of 185/333 genes. Furthermore, animals may share a suite of 185/333 genes that are expressed in response to common pathogens, while also maintaining a large number of unique genes within the population