Dissection of Nodule Development by Supplementation of \u3cem\u3eRhizobium leguminosarum\u3c/em\u3e biovar \u3cem\u3ephaseoli\u3c/em\u3e Purine Auxotrophs with 4-Aminoimidazole-5-Carboxamide Riboside

Purine auxotrophs of Rhizobium leguminosarum biovar phaseoli CFN42 elicit uninfected pseudonodules on bean (Phaseolus vulgaris L.). Addition of 4-aminoimidazole-5-carboxamide (AICA) riboside to the root medium during incubation of the plant with these mutants leads to enhanced nodule development, although nitrogenase activity is not detected. Nodules elicited in this manner had infection threads and anatomical features characteristic of normal nodules, such as peripheral vasculature rather than the central vasculature of the pseudonodules that were elicited without AICA riboside supplementation. Although 105 to 106 bacteria could be recovered from these nodules after full development, bacteria were not observed in the interior nodule cells. Instead, large cells with extensive internal membranes were present. Approximately 5% of the normal amount of leghemoglobin and 10% of the normal amount of uricase were detected in these nodules. To promote the development of true nodules rather than pseudonodules, AICA riboside was required no later than the second day through no more than the sixth day following inoculation. After this period, removal of AICA riboside from the root medium did not prevent the formation of true nodules. This observation suggests that there is a critical stage of infection, reached before nodule emergence, at which development becomes committed to forming a true nodule rather than a pseudonodule

Spatial and Behavioral Patterns of Captive Coyotes

Environmental enrichment is a technique used at many captive animal facilities that can improve the well-being of their animals. It seeks to enhance habitat features and promote natural behavior by providing a variety of practical ways for captive animals to control their environmental settings, especially during stressful circumstances. Enclosure features, such as shelter structures, are one tool that promotes wild behavior by adding complexity to an enclosure’s physical environment. Enrichment efforts for captive wildlife are most effective when they are specialized to the biological needs of the animals. Human activity may alter captive animal behavior and utility of enclosure features, and there is concern that human presence can negatively impact the welfare of some captive animals. Captive coyotes (Canis latrans) at the USDA-National Wildlife Research Center (NWRC) Predator Research Facility in Millville, UT, USA, are maintained for research on biology, ecology, physiology and behavior. Coyotes at the research facility are routinely noticed utilizing shelter structures to hide, rest, and display vigilant behavior. Because they regularly use these simple structures, new and more complex enrichment shelter structures were installed to be evaluated. Specific research objectives aimed to assess (1) coyote enclosure utilization and shelter structure preferences, and (2) coyote spatial and behavioral responses to human activity. Using 32 mated coyote pairs rotated through eight 1.5-acre enclosures for 28-day trials over the winter months (January – March) of 2015 and 2016, spatial and behavioral patterns were monitored via the implementation of GPS-collars and live behavioral observations. Coyotes showed preference for shelter structure designs, but still spent most of their time at the perimeter and open areas of their enclosures. Complex structures were preferred over simple structures. Coyotes most often demonstrated inactive and vigilant behavior without human activity, but showed increased vigilance when there was human activity. Human activity also stimulated coyotes to become more active than inactive and reduce their utilization of enrichment structures. Although there was no clear preference for one specific type of enrichment structure, composite evidence from GPS-collars and behavioral data suggest the ramp may have heightened biological suitability. This study advances the knowledge of captive coyote spatial patterns and helps improve environmental enrichment planning for captive animals by exploring effective methods of adding complexity to animal enclosures

Infection of Soybean and Pea Nodules by \u3cem\u3eRhizobium\u3c/em\u3e spp. Purine Auxotrophs in the Presence of 5-aminoimidazole-4-Carboxamide Riboside

Purine auxotrophs of various Rhizobium species are symbiotically defective, usually unable to initiate or complete the infection process. Earlier studies demonstrated that, in the Rhizobium etli-bean symbiosis, infection by purine auxotrophs is partially restored by supplementation of the plant medium with 5-amino-imidazole-4-carboxamide (AICA) riboside, the unphosphorylated form of the purine biosynthetic intermediate AICAR. The addition of purine to the root environment does not have this effect. In this study, purine auxotrophs of Rhizobium fredii HH303 and Rhizobium leguminosarum 128C56 (bv. viciae) were examined. Nutritional and genetic characterization indicated that each mutant was blocked in purine biosynthesis prior to the production of AICAR. R. fredii HH303 and R. leguminosarum 128C56 appeared to be deficient in AICA riboside transport and/or conversion into AICAR, and the auxotrophs derived from them grew very poorly with AICA riboside as a purine source. All of the auxotrophs elicited poorly developed, uninfected nodules on their appropriate hosts. On peas, addition of AICA riboside or purine to the root environment led to enhanced nodulation; however, infection threads were observed only in the presence of AICA riboside. On soybeans, only AICA riboside was effective in enhancing nodulation and promoting infection. Although AICA riboside supplementation of the auxotrophs led to infection thread development on both hosts, the numbers of bacteria recovered from the nodules were still 2 or more orders of magnitude lower than in fully developed nodules populated by wild-type bacteria. The ability to AICA riboside to promote infection by purine auxotrophs, despite serving as a very poor purine source for these strains, supports the hypothesis that AICAR plays a role in infection other than merely promoting bacterial growth

Are Four-Year Universities Better Than Two-Year Colleges at Preparing Students to Pass the FAA Aircraft Mechanic Certification Written Examinations?

This study evaluates differences between two-year and four-year schools offering a Federal Aviation Administration (FAA) Part 147 aviation maintenance technician program. The actual average test scores were analyzed to determine whether students from four-year university programs were better prepared for the test than students from two-year college or technical school programs. Test scores can be viewed as a direct indication of the quality of the aviation maintenance programs at these schools. The study found that there was no statistically significant difference in the test scores between two-year college and four-year university students taking the FAA examinations. These results support previous studies that found the learning outcomes of two-year and four-year students on general subjects to be comparable

Identification of Unknown Contaminants in Water Samples from ISS Employing Liquid Chromatography/Mass Spectrometry/Mass Spectrometry

Mass Spectrometry/Mass Spectrometry (MS/MS) is a powerful technique for identifying unknown organic compounds. For non-volatile or thermally unstable unknowns dissolved in liquids, liquid chromatography/mass spectrometry/mass spectrometry (LC/MS/MS) is often the variety of MS/MS used for the identification. One type of LC/MS/MS that is rapidly becoming popular is time-of-flight (TOF) mass spectrometry. This technique is now in use at the Johnson Space Center for identification of unknown nonvolatile organics in water samples from the space program. An example of the successful identification of one unknown is reviewed in detail in this paper. The advantages of time-of-flight instrumentation are demonstrated through this example as well as the strategy employed in using time-of-flight data to identify unknowns

Implant Prophylaxis: The Next Best Practice Toward Asepsis in Spine Surgery.

Study designA literature review.ObjectivesAn evaluation of the contaminants prevalent on implants used for surgery and the aseptic methods being employed against them.MethodsPubMed was searched for articles published between 2000 and 2017 for studies evaluating the contaminants present on spine implants, and associated pre- and intraoperative implant processing and handling methodology suggested to avoid them. Systematic reviews, observational studies, bench-top studies, and expert opinions were included.ResultsEleven studies were identified whose major focus was the asepsis of implants to reduce the incidence of surgical site infection incidences during surgery. These studies measured the colony forming units of bacteria on sterilized implants and/or gloves from the surgeon, scrub nurse, and assistants, as well as reductions of surgical site infection rates in spine surgery due to changes in implant handling techniques. Additionally, the search included assessments of endotoxins and carbohydrates present on reprocessed implants. The suggested changes to surgical practice based on these studies included handling implants with only fresh gloves, keeping implants covered until the immediate time of use, reducing operating room traffic, avoiding reprocessing of implants (ie, providing terminally sterilized implants), and avoiding touching the implants altogether.ConclusionsBoth reprocessing (preoperative) and handling (intraoperative) of implants seem to lead to contamination of sterilized implants. Using a terminally sterilized device may mitigate reprocessing (preoperative implant prophylaxis), whereas the use of fresh gloves for handling each implant and/or a permanent shielding technique (intraoperative implant prophylaxis) could potentially avoid recontamination at the theatre

Atmospheric circulation patterns, cloud-to-ground lightning, and locally intense convective rainfall associated with debris flow initiation in the Dolomite Alps of northeastern Italy

The Dolomite Alps of northeastern Italy experience debris flows with great frequency during the summer months. An ample supply of unconsolidated material on steep slopes and a summer season climate regime characterized by recurrent thunderstorms combine to produce an abundance of these destructive hydro-geologic events. In the past, debris flow events have been studied primarily in the context of their geologic and geomorphic characteristics. The atmospheric contribution to these mass-wasting events has been limited to recording rainfall and developing intensity thresholds for debris mobilization. This study aims to expand the examination of atmospheric processes that preceded both locally intense convective rainfall (LICR) and debris flows in the Dolomite region. 500 hPa pressure level plots of geopotential heights were constructed for a period of 3 days prior to debris flow events to gain insight into the synoptic-scale processes which provide an environment conducive to LICR in the Dolomites. Cloud-to-ground (CG) lightning flash data recorded at the meso-scale were incorporated to assess the convective environment proximal to debris flow source regions. Twelve events were analyzed and from this analysis three common synoptic-scale circulation patterns were identified. Evaluation of CG flashes at smaller spatial and temporal scales illustrated that convective processes vary in their production of CF flashes (total number) and the spatial distribution of flashes can also be quite different between events over longer periods. During the 60 min interval immediately preceding debris flow a majority of cases exhibited spatial and temporal colocation of LICR and CG flashes. Also a number of CG flash parameters were found to be significantly correlated to rainfall intensity prior to debris flow initiation

Wildlife responses to livestock guard dogs and domestic sheep on open range

Domestic sheep (Ovis aries) are an attractant to carnivores; however, sheep are often accompanied by humans and livestock guardian dogs (LGDs; Canis familiaris), which defend sheep from depredation. Sheep also compete directly with wildlife for grazing resources. Although practiced for millennia in much of the world outside North America, the effect that transhumance has on wildlife is poorly understood. To test the effect of sheep bands (sheep, humans, and LGDs) on wildlife, we modeled the detection probability of wild mammals relative to the presence of sheep bands in the Northwestern United States. Sheep band presence was associated with a reduction of about half in the likelihood of detecting large carnivores (Ursus americanus, Ursus arctos, Canis lupus, and Puma concolor, p \u3c 0.05) and deer (Odocoileus spp., p \u3c 0.01), both while the band was present and after it left the area. Contrastingly, coyotes (Canis latrans) were more than three times as likely to be detected when sheep bands were present (p \u3c 0.001), and twice as likely after sheep bands left (p \u3c 0.01). Coyotes were the only species we modeled that was more likely to be detected when a sheep band was present. It is unclear how long these effects persist after a sheep band has moved through an area, but our results suggest that transhumance temporarily displaces many large mammals, which results in mesopredator release of coyotes. This study suggests there is a tradeoff between the conservation benefits provided by LGDs and humans protecting sheep and the costs of displacement to some wild mammals

Structure of the RNA-dependent RNA polymerase of poliovirus

AbstractBackground: The central player in the replication of RNA viruses is the viral RNA-dependent RNA polymerase. The 53 kDa poliovirus polymerase, together with other viral and possibly host proteins, carries out viral RNA replication in the host cell cytoplasm. RNA-dependent RNA polymerases comprise a distinct category of polymerases that have limited sequence similarity to reverse transcriptases (RNA-dependent DNA polymerases) and perhaps also to DNA-dependent polymerases. Previously reported structures of RNA-dependent DNA polymerases, DNA-dependent DNA polymerases and a DNA-dependent RNA polymerase show that structural and evolutionary relationships exist between the different polymerase categories.Results: We have determined the structure of the RNA-dependent RNA polymerase of poliovirus at 2.6 Å resolution by X-ray crystallography. It has the same overall shape as other polymerases, commonly described by analogy to a right hand. The structures of the ‘fingers’ and ‘thumb’ subdomains of poliovirus polymerase differ from those of other polymerases, but the palm subdomain contains a core structure very similar to that of other polymerases. This conserved core structure is composed of four of the amino acid sequence motifs described for RNA-dependent polymerases. Structure-based alignments of these motifs has enabled us to modify and extend previous sequence and structural alignments so as to relate sequence conservation to function. Extensive regions of polymerase–polymerase interactions observed in the crystals suggest an unusual higher order structure that we believe is important for polymerase function.Conclusions: As a first example of a structure of an RNA-dependent RNA polymerase, the poliovirus polymerase structure provides for a better understanding of polymerase structure, function and evolution. In addition, it has yielded insights into an unusual higher order structure that may be critical for poliovirus polymerase function