Localization of the paranodal protein Caspr in the mammalian retina

Purpose: The retina has the demanding task of encoding all aspects of the visual scene within the space of one fixation period lasting only a few hundred milliseconds. To accomplish this feat, information is encoded in specialized parallel channels and passed on to numerous central nuclei via the optic nerve. These parallel channels achieve specialization in at least three ways: the synaptic networks in which they participate, the neurotransmitter receptors expressed and the types and locations of ion channels or transporters used. Subcellular localization of receptors, channels and transporters is made yet more complex in the retina by the double duty many retinal processes serve. In the present work, we show that the protein Caspr (Contactin Associated Protein), best known for its critical role in the localization of voltage-gated ion channels at the nodes of Ranvier, is present in several types of retinal neurons including amacrine, bipolar, horizontal, and ganglion cells. Methods: Using standard double label immunofluorescence protocols, we characterized the pattern of Caspr expression in the rodent retina. Results: Caspr labeling was observed through much of the retina, including horizontal, bipolar, amacrine, and ganglion cells. Among amacrine cells, Caspr was observed in AII amacrine cells through co-localization with Parvalbumin and Disabled-1 in rat and mouse retinas, respectively. An additional amacrine cell type containing Calretinin also co-localized with Caspr, but did not co-localize with choline-acetyltransferase. Nearly all cells in the ganglion cell layer contain Caspr, including both displaced amacrine and ganglion cells. In the outer retina, Caspr was co-localized with PKC labeling in rod bipolar cell dendrites. In addition, Caspr labeling was found inside syntaxin-4 'sandwiches' in the outer plexiform layer, most likely indicating its presence in cone bipolar cell dendrites. Finally, Caspr was co-localized in segments of horizontal cell dendrites labeled with Calbindin-D28k. Conclusions: Caspr is best known for its role in organizing the localization of different voltage-gated ion channels in and around nodes of Ranvier. As neuronal processes in the retina often play a dual role involving both input and output, it is possible that the localization of Caspr in the retina will help us decipher the way retinal cells localize ion channels in their processes to increase computational capacity

Southern California Regional Workforce Development Needs Assessment for the Transportation and Supply Chain Industry Sectors

COVID-19 brought the public’s attention to the critical value of transportation and supply chain workers as lifelines to access food and other supplies. This report examines essential job skills required of the middle-skill workforce (workers with more than a high school degree, but less than a four-year college degree). Many of these middle-skill transportation and supply chain jobs are what the Federal Reserve Bank defines as “opportunity occupations” -- jobs that pay above median wages and can be accessible to those without a four-year college degree. This report lays out the complex landscape of selected technological disruptions of the supply chain to understand the new workforce needs of these middle-skill workers, followed by competencies identified by industry. With workplace social distancing policies, logistics organizations now rely heavily on data management and analysis for their operations. All rungs of employees, including warehouse workers and truck drivers, require digital skills to use mobile devices, sensors, and dashboards, among other applications. Workforce training requires a focus on data, problem solving, connectivity, and collaboration. Industry partners identified key workforce competencies required in digital literacy, data management, front/back office jobs, and in operations and maintenance. Education and training providers identified strategies to effectively develop workforce development programs. This report concludes with an exploration of the role of Institutes of Higher Education in delivering effective workforce education and training programs that reimagine how to frame programs to be customizable, easily accessible, and relevant

Risk Factors and Outcome of Fontan‐Associated Plastic Bronchitis: A Case‐Control Study

Peer Reviewedhttps://deepblue.lib.umich.edu/bitstream/2027.42/139068/1/jah3521.pd

Yeast require redox switching in DNA primase

Eukaryotic DNA primases contain a [4Fe4S] cluster in the C-terminal domain of the p58 subunit (p58C) that affects substrate affinity but is not required for catalysis. We show that, in yeast primase, the cluster serves as a DNA-mediated redox switch governing DNA binding, just as in human primase. Despite a different structural arrangement of tyrosines to facilitate electron transfer between the DNA substrate and [4Fe4S] cluster, in yeast, mutation of tyrosines Y395 and Y397 alters the same electron transfer chemistry and redox switch. Mutation of conserved tyrosine 395 diminishes the extent of p58C participation in normal redox-switching reactions, whereas mutation of conserved tyrosine 397 causes oxidative cluster degradation to the [3Fe4S]^+ species during p58C redox signaling. Switching between oxidized and reduced states in the presence of the Y397 mutations thus puts primase [4Fe4S] cluster integrity and function at risk. Consistent with these observations, we find that yeast tolerate mutations to Y395 in p58C, but the single-residue mutation Y397L in p58C is lethal. Our data thus show that a constellation of tyrosines for protein-DNA electron transfer mediates the redox switch in eukaryotic primases and is required for primase function in vivo

Meeting Report: The 2nd Annual Argonne Soils Workshop, Argonne National Laboratory, Chicago Illinois, USA, October 6-8, 2010

This report summarizes the proceedings of the 2nd Annual Argonne Soils Workshop held at Argonne National Laboratory October 6–8, 2010. The workshop assembled a diverse group of soil ecologists, microbiologists, molecular biologists, and computational scientists to discuss the challenges and opportunities related to implementation of metagenomics approaches in soil microbial ecology. The overarching theme of the workshop was “designing ecologically meaningful soil metagenomics research”, which encouraged presentations on both ecological and computational topics. The workshop fostered valuable cross-discipline communication and delivered the message that soil metagenomics research must be based on an iterative process between biological inquiry and bioinformatics tools

Familial Aggregation of Survival and Late Female Reproduction

Abstract not availabl

Investigation of free-living honey bee colonies in Ireland

Apis mellifera mellifera (Linnaeus), the Western European honey bee, is considered extinct in the wild over most of its range due largely to hybridisation and replacement by other subspecies, parasitism by Varroa destructor, habitat loss, and effects from agricultural pesticides. The purity of the subspecies within the managed cohort is also at risk over much of its range. Here, we investigated if honey bee colonies inhabited locations outside of the apiaries. In those we located, we explored how long the colony persisted and we investigated the genotypes of the bees using multiple markers. We show here that unmanaged free-living honey bee colonies are present and widespread in Ireland, inhabiting a mixture of nesting habitats with some colonies persisting naturally and unaided over multiple years. Molecular data including mitochondrial, microsatellite, and SNPs evidence indicate that the free-living population sampled is largely comprised of pure A. m. mellifera. Finally, we discuss the implications of conserving free-living A. m. mellifera in Ireland and its possible role in improving the fitness of the managed population both in Ireland and the rest of its European range.We particularly thank the custodians of the free-living honey bee colonies and the Native Irish Honey Bee Society (NIHBS) for their assistance. KAB is a recipient of an Irish Research Council postgraduate fellowship (GOIPG/2015/2767) and a Tony Ryan Postgraduate fellowship. Additional funding was gratefully received from the Department of Agriculture, Food and the Marine [grant number GRGAS 16/GR/09], the Federation of Irish Beekeeping Associations, the Eva Crane Trust [grant number ECTA20160303] and The Native Irish Honey Bee Society. Financial support for DH was provided through the program COMPETE 2020 – POCI (Programa Operacional para a Competividade e Internacionalizac¸~ao) and by Portuguese funds through FCT (Fundac¸~ao para a Ci^encia e a Tecnologia) in the framework of the project BeeHappy (POCI-01-0145-FEDER-029871).info:eu-repo/semantics/publishedVersio

The Time is Right for an Antarctic Biorepository Network

Antarctica is a central driver of the Earth’s climate and health. The Southern Ocean surrounding Antarctica serves as a major sink for anthropogenic CO2 and heat (1), and the loss of Antarctic ice sheets contributes significantly to sea level rise and will continue to do so as the loss of ice sheets accelerates, with sufficient water stores to raise sea levels by 58 m (2). Antarctica\u27s marine environment is home to a number of iconic species, and the terrestrial realm harbors a remarkable oasis for life, much of which has yet to be discovered (3). Distinctive oceanographic features of the Southern Ocean—including the Antarctic Circumpolar Current, the Antarctic Polar Front, and exceptional depths surrounding the continent—coupled with chronically cold temperatures have fostered the evolution of a vast number of uniquely coldadapted species, many of which are found nowhere else on the Earth (4). The Antarctic marine biota, for example, displays the highest level of species endemism on the Earth (5). However, warming, ocean acidification, pollution, and commercial exploitation threaten the integrity of Antarctic ecosystems (6). Understanding changes in the biota and its capacities for adaptation is imperative for establishing effective policies for mitigating the impacts of climate change and sustaining the Antarctic ecosystems that are vital to global health

Functional and structural similarity of human DNA primase [4Fe4S] cluster domain constructs

The regulatory subunit of human DNA primase has a C-terminal domain (p58C) that contains a [4Fe4S] cluster and binds DNA. Previous electrochemical analysis of a p58C construct revealed that its affinity for DNA is sensitive to the redox state of the [4Fe4S] cluster. Concerns about the validity of this conclusion have been raised, based in part on differences in X-ray crystal structures of the p58C_(272-464) construct used for that study and that of a N-terminally shifted p58C_(266-456) construct and consequently, an assumption that p58C_(272-464) has abnormal physical and functional properties. To address this controversy, a new p58C_(266-464) construct containing all residues was crystallized under the conditions previously used for crystallizing p58C_(272-464), and the solution structures of both constructs were assessed using circular dichroism and NMR spectroscopy. In the new crystal structure, p58C_(266-464) exhibits the same elements of secondary structure near the DNA binding site as observed in the crystal structure of p58C_(272-464). Moreover, in solution, circular dichroism and ^(15)N,^1H-heteronuclear single quantum coherence (HSQC) NMR spectra show there are no significant differences in the distribution of secondary structures or in the tertiary structure or the two constructs. To validate that the two constructs have the same functional properties, binding of a primed DNA template was measured using a fluorescence-based DNA binding assay, and the affinities for this substrate were the same (3.4 ± 0.5 μM and 2.7 ± 0.3 μM, respectively). The electrochemical properties of p58C_(266-464) were also measured and this p58C construct was able to engage in redox switching on DNA with the same efficiency as p58C_(272-464). Together, these results show that although p58C can be stabilized in different conformations in the crystalline state, in solution there is effectively no difference in the structure and functional properties of p58C constructs of different lengths