Draft Genome Sequences of Four Saccharibacter sp. Strains Isolated from Native Bees.

The genus Saccharibacter is currently understudied, with only one described species, Saccharibacter floricola, isolated from a flower. In an effort to better understand the microbes that come in contact with native bee pollinators, we isolated and sequenced four additional strains of Saccharibacter from native bees in the genera Melissodes and Anthophora These genomes range in size from 2,104,494 to 2,316,791 bp (mean, 2,246,664 bp) and contain between 1,860 and 2,167 (mean, 2,060) protein-coding genes

Testing of Noodle Creek Site 41JS102 Jones County, Texas

Staff Archeologists from the Environmental Affairs Division of Texas Department of Transportation discovered buried cultural resources that constitute archeological site 41JS102, in June and July 2001 at the proposed location of a bridge replacement over Noodle Creek in Jones County, Texas. Shovel testing and backhoe trenching revealed cultural material at 65 cm below the surface. Following this discovery the Texas Department of Transportation made recommendations to the Texas Historical Commission to assess site 41JS 102 and they concurred. In September 2001 archeologists from the Texas Department of Transportation initiated the site assessment phase. Landowner complications halted that work before completion. Subsequently, TRC Environmental was provided a Work Authorization under an existing general services contract to complete the assessment of 41JS102 in December 2001. The fieldwork consisted of the hand excavation and documentation of eight 1 by 1 m test units and the digging and documenting of six backhoe trenches. Limited and scattered historic artifacts were recovered from the upper parts of the two alluvial terraces (T0 and T1). A prehistoric component with quantities of lithic debitage and sparse burned rocks, limited stone tools and mussel shells, and one possible cultural feature were encountered in a sloping, but well defined 10 to 15 cm thick zone in the T1 deposits. The Principal Investigator recommended prehistoric site 4 1JS102 was eligible for inclusion in the National Register of Historic Places and as a State Archeological Landmark. However, the staff at Texas Department of Transportation disagreed with that recommendation and presented an opposing case to the Texas Historical Commission who concurred with the Texas Department of Transportation\u27s recommendations. Consequently, no further archeological investigations will occur prior to the proposed development

Failure to communicate: Inefficiencies in voluntary incentive programs for private forest owners in Michigan

Coordinating forest management across thousands of nonindustrial private forest (NIPF) owners is a difficult yet necessary task for state land management agencies. Voluntary Incentive Programs (VIPs) can coordinate the decentralized activities of these owners in return for services or financial incentives. However, many VIPs typically have low enrollment. Our study investigates the implementation of VIPs to increase forest management coordination among NIPFs in Michigan. We present findings from 20 semi-structured interviews with leaders of state and local land management organizations, and government officials at state natural resource agencies, and contrast their answers with those recorded from 37 interviews of NIPF owners regarding their knowledge and attitudes toward VIPs. Our interviews highlight a critical disconnect between NIPF owner motivations and VIP incentives, as well as misallocated resources for VIP promotion by state agencies, driving low enrollment. At the core, low enrollment in VIPs is generated by inadequate communication between NIPF owners and program managers, along with distrust of government agency objectives. Viewing managers as “street level bureaucrats”, civil servants whose job discretion is impacted heavily by available resources, may increase our understanding of the issues plaguing VIPs and help identify improvements to VIP design and implementation

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

Aggregation of Sea Urchin Phagocytes Is Augmented In Vitro by Lipopolysaccharide

Development of protocols and media for culturing immune cells from marine invertebrates has not kept pace with advancements in mammalian immune cell culture, the latter having been driven by the need to understand the causes of and develop therapies for human and animal diseases. However, expansion of the aquaculture industry and the diseases that threaten these systems creates the need to develop cell and tissue culture methods for marine invertebrates. Such methods will enable us to better understand the causes of disease outbreaks and to develop means to avoid and remedy epidemics. We report a method for the short-term culture of phagocytes from the purple sea urchin, Strongylocentrotus purpuratus, by modifying an approach previously used to culture cells from another sea urchin species. The viability of cultured phagocytes from the purple sea urchin decreases from 91.6% to 57% over six days and phagocyte morphology changes from single cells to aggregates leading to the formation of syncytia-like structures. This process is accelerated in the presence of lipopolysaccharide suggesting that phagocytes are capable of detecting this molecular pattern in culture conditions. Sea urchin immune response proteins, called Sp185/333, are expressed on the surface of a subset of phagocytes and have been associated with syncytia-like structures. We evaluated their expression in cultured phagocytes to determine their possible role in cell aggregation and in the formation of syncytia-like structures. Between 0 and 3 hr, syncytia-like structures were observed in cultures when only similar to 10% of the cells were positive for Sp185/333 proteins. At 24 hr, similar to 90% of the nuclei were Sp185/333-positive when all of the phagocytes had aggregated into syncytia-like structures. Consequently, we conclude that the Sp185/333 proteins do not have a major role in initiating the aggregation of cultured phagocytes, however the Sp185/333 proteins are associated with the clustered nuclei within the syncytia-like structures

Teacher-made Tactile Science Materials with Critical and Creative Thinking Activities for Learners Including those with Visual Impairments

Gifted students with visual impairments are twice exceptional learners and may not evidence their advanced science aptitudes without appropriate accommodations for learning science. However, effective tactile science teaching materials may be easily made. Recent research has shown that when tactile materials are used with all students in the class, everyone benefits. This presentation provides many classroom-tested example sets of tactile materials for teaching upper elementary and middle school science concepts. These science concepts include the parts of: a flower, ear, insect, beaver habitat, volcano, hydroelectric power plant, plant cell, and sun. Critical and creative thinking skill strategies to accompany these materials for further developing gifted students’ science knowledge also are provided. These include application of the Edward de Bono CoRT Breadth thinking skills and other creative thinking skills, such as making a model or using analogy. This document represents the content of a gifted education conference presentation made at the Iowa Talented and Gifted Association Annual Meeting

Department of Energy Solar Decathlon 2015

The goal of the Department of Energy (DOE) Solar Decathlon 2015 project is to design, build, and operate a zero-net energy solar-photovoltaic (solar-PV) powered residence. It is an international competition taking place in the fall of 2015 in Irvine, California. The design needs to be attractive, energy-efficient, innovative, and affordable. The Cal Poly team includes students from a variety of majors, including Architecture, Mechanical Engineering, Electrical Engineering, and many more. The different disciplines need to work together to create a functional and cohesive design. For our senior project, we have designed the electrical systems in the home, including the power, lighting, solar-PV systems, and energy monitoring. The design flow has four stages, Schematic Design (SD), Design Documents (DD), Construction Documents (CD), and Construction Administration (CA). Models were created using the AutoDesk software Revit, which is a building information modeling program with real-time 3D rendering. Our electrical design must comply with state and national building codes, such as the California Electrical Code (CEC) and National Electrical Code (NEC)

Single-strand nicks induce homologous recombination with less toxicity than double-strand breaks using an AAV vector template

Gene targeting by homologous recombination (HR) can be induced by double-strand breaks (DSBs), however these breaks can be toxic and potentially mutagenic. We investigated the I-AniI homing endonuclease engineered to produce only nicks, and found that nicks induce HR with both plasmid and adeno-associated virus (AAV) vector templates. The rates of nick-induced HR were lower than with DSBs (24-fold lower for plasmid transfection and 4- to 6-fold lower for AAV vector infection), but they still represented a significant increase over background (240- and 30-fold, respectively). We observed severe toxicity with the I-AniI ‘cleavase’, but no evidence of toxicity with the I-AniI ‘nickase.’ Additionally, the frequency of nickase-induced mutations at the I-AniI site was at least 150-fold lower than that induced by the cleavase. These results, and the observation that the surrounding sequence context of a target site affects nick-induced HR but not DSB-induced HR, strongly argue that nicks induce HR through a different mechanism than DSBs, allowing for gene correction without the toxicity and mutagenic activity of DSBs

Insect Cell Expression and Purification of Recombinant SARS-COV-2 Spike Proteins That Demonstrate ACE2 Binding

The COVID-19 pandemic caused by SARS-CoV-2 infection has led to socio-economic shutdowns and the loss of over 5 million lives worldwide. There is a need for the identification of therapeutic targets to treat COVID-19. SARS-CoV-2 spike is a target of interest for the development of therapeutic targets. We developed a robust SARS-CoV-2 S spike expression and purification protocol from insect cells and studied four recombinant SARS-CoV-2 spike protein constructs based on the original SARS-CoV-2 sequence using a baculovirus expression system: a spike protein receptor-binding domain that includes the SD1 domain (RBD) coupled to a fluorescent tag (S-RBD-eGFP), spike ectodomain coupled to a fluorescent tag (S-Ecto-eGFP), spike ectodomain with six proline mutations and a foldon domain (S-Ecto-HexaPro(+F)), and spike ectodomain with six proline mutations without the foldon domain (S-Ecto-HexaPro(-F)). We tested the yield of purified protein expressed from the insect cell lines Spodoptera frugiperda (Sf9) and Trichoplusia ni (Tni) and compared it to previous research using mammalian cell lines to determine changes in protein yield. We demonstrated quick and inexpensive production of functional glycosylated spike protein of high purity capable of recognizing and binding to the angiotensin converting enzyme 2 (ACE2) receptor. To further confirm functionality, we demonstrate binding of eGFP fused construct of the spike ectodomain (S-Ecto-eGFP) to surface ACE2 receptors on lung epithelial cells by flow cytometry analysis and show that it can be decreased by means of receptor manipulation (blockade or downregulation)

Discovery of a Dynamical Cold Point in the Heart of the Sagittarius dSph Galaxy with Observations from the APOGEE Project

The dynamics of the core of the Sagittarius (Sgr) dwarf spheroidal (dSph) galaxy are explored using high-resolution (R~22,500), H-band, near-infrared spectra of over 1,000 giant stars in the central 3 deg^2 of the system, of which 328 are identified as Sgr members. These data, among some of the earliest observations from the SDSS-III/Apache Point Observatory Galactic Evolution Experiment (APOGEE) and the largest published sample of high resolution Sgr dSph spectra to date, reveal a distinct gradient in the velocity dispersion of Sgr from 11-14 km/s for radii >0.8 degrees from center to a dynamical cold point of 8 km/s in the Sgr center --- a trend differing from that found in previous kinematical analyses of Sgr over larger scales that suggest a more or less flat dispersion profile at these radii. Well-fitting mass models with either cored and cusped dark matter distributions can be found to match the kinematical results, although the cored profile succeeds with significantly more isotropic stellar orbits than required for a cusped profile. It is unlikely that the cold point reflects an unusual mass distribution. The dispersion gradient may arise from variations in the mixture of populations with distinct kinematics within the dSph; this explanation is suggested (e.g., by detection of a metallicity gradient across similar radii), but not confirmed, by the present data. Despite these remaining uncertainties about their interpretation, these early test data (including some from instrument commissioning) demonstrate APOGEE's usefulness for precision dynamical studies, even for fields observed at extreme airmasses.Comment: 15 pages, 3 figure