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At least 19 records

Composition and physiological profiling of sprout-associated microbial communities

The native microfloras of various types of sprouts (alfalfa, clover, sunflower, mung bean, and broccoli sprouts) were examined to assess the relative effects of sprout type and inoculum factors (i.e., sprout-growing facility, seed lot, and inoculation with sprout-derived inocula) on the microbial community structure of sprouts. Sprouts were sonicated for 7 min or hand shaken with glass beads for 2 min to recover native microfloras from the surface, and the resulting suspensions were diluted and plated. The culturable fraction was characterized by the density (log CFU/g), richness (e.g., number of types of bacteria), and diversity (e.g., microbial richness and evenness) of colonies on tryptic soy agar plates incubated for 48 h at 30 degrees C. The relative similarity between sprout-associated microbial communities was assessed with the use of community-level physiological profiles (CLPPs) based on patterns of utilization of 95 separate carbon sources. Aerobic plate counts of 7.96 +/- 0.91 log CFU/g of sprout tissue (fresh weight) were observed, with no statistically significant differences in microbial cell density, richness, or diversity due to sprout type, sprout-growing facility, or seed lot. CLPP analyses revealed that the microbial communities associated with alfalfa and clover sprouts are more similar than those associated with the other sprout types tested. Variability among sprout types was more extensive than any differences between microbial communities associated with alfalfa and clover sprouts from different sprout-growing facilities and seed lots. These results indicate that the subsequent testing of biocontrol agents should focus on similar organisms for alfalfa and clover, but alternative types may be most suitable for the other sprout types tested. The inoculation of alfalfa sprouts with communities derived from various sprout types had a significant, source-independent effect on microbial community structure, indicating that the process of inoculation alters the dynamics of community development regardless of the types of organisms involved.

NASA Center KSC↗

The water factor in harvest-sprouting of hard red spring wheat

Sprouting in unthreshed, ripe, hard red spring wheat (Triticum aestivum L.) is induced by rain, but sprouting does not necessarily occur because the crop is wetted. The spike and grain water conditions conducive to sprouting were determined in a series of laboratory experiments. Sprouting did not occur in field growing wheat wetted to 110% water concentration until the spike water concentration was reduced to 12% and maintained at this concentration for 2 days before wetting. When cut at growth stage 11.3, Feekes scale, Saratovskaya 20 (USSR) sprouted after 4 days drying, Olaf and Alex between 7 and 15 days drying and Columbus, recognized for its resistance to harvest time sprouting, after more than 15 days drying. Sprouting potential was enhanced after 4 wetting drying cycles in which any wetted interval was too brief to permit sufficient water imbibition to initiate sprouting. At harvest ripeness, grain water concentration exceeded spike water concentration by 0.7 percentage units. Following 6 months storage, 20% of the kernels in 300 spike bundles (simulating windrows) sprouted within 28 hrs after initiation of wetting to saturation (150% water concentration). Ninety percent sprouting occurred within 8 days in bundles maintained at 75% water concentration and higher, but less sprouting occurred in bundles dried to 50% water concentration before resaturation.

Bauer, A.↗

Seed sprout production: Consumables and a foundation for higher plant growth in space

Seed sprouts can be produced as a source of fresh vegetable materials and as higher plant seedlings in space. Sprout production was undertaken to evaluate the mass accumulations possible, the technologies needed, and the reliability of the overall process. Baseline experiments corroborated the utility of sprout production protocols for a variety of seed types. The automated delivery of saturated humidity effectively supplants labor intensive manual soaking techniques. Automated humidification also lend itself to modest centrifugal sprout growth environments. A small amount of ultraviolet radiation effectively suppressed bacterial and fungal contamination, and the sprouts were suitable for consumption.

Day, Michelle↗

Automatic Sprout Grower

Self-contained seed-sprouting system provides environment for sprouting seeds quickly and easily. Sprouting container standard 6-oz package for dehydrated food and drink mixes in Space Shuttle. About 4 g of dry alfalfa or radish seeds vacuum-sealed in each cup, like freeze-dried foods. Sixteen cups suspended in tray. Air-and-water inlet tube links each cup to system of tubes and solenoid valves alternately furnish air and water and remove stale air. Peristaltic pump supplies water from vinyl medical-fluid bag. Small diaphragm pump supplies and exhausts air. Small circuit board times movements of air and water. Kit offers advantages to home gardeners. Apartment dwellers use it for steady production of homegrown sprouts even though they have no garden space.

Sauer, Richard L.↗

Environmental factors during seed development and their influence on pre-harvest sprouting in wheat

The problem of pre-harvest sprouting of wheat is surveyed and a literature review of the effects of environmental conditions on pre-harvest sprouting is presenting. Physiological, biochemical, and morphological changes occurring within the wheat seed during germination, harvest, and storage are discussed. The effects of moisture, humidity, and temperature, particularly on seed dormancy, are considered. Procedures used in Europe for predicting the potential for sprouting are evaluated.

Ciha, A. J.↗

By Different Cellular Mechanisms, Lymphatic Vessels Sprout by Endothelial Cell Recruitment Whereas Blood Vessels Grow by Vascular Expansion

The development of effective vascular therapies requires the understanding of all modes of vessel formation contributing to vasculogenesis, angiogenesis (here termed hemangiogenesis) and lymphangiogenesis. We show that lymphangiogenesis proceeds by blind-ended vessel sprouting via recruitment of isolated endothelial progenitor cells to the tips of growing vessels, whereas hemangiogenesis occurs by non-sprouting vessel expansion from the capillary network, during middevelopment in the quail chorioallantoic membrane (CAM). Blood vessels expanded out of capillaries that displayed transient expression of alpha smooth muscle actin (alphaSMA), accompanied by mural recruitment of migratory progenitor cells expressing SMA. Lymphatics and blood vessels were identified by confocal/fluorescence microscopy of vascular endothelial growth factor (VEGF) receptors VEGFR-1 and VEGFR-2, alphaSMA (expressed on CAM blood vessels but not on lymphatics), homeobox transcription factor Prox-1 (specific to CAM lymphatic endothelium), and the quail hematopoetic/vascular marker, QH-1. Expression of VEGFR-1 was highly restricted to blood vessels (primarily capillaries). VEGFR-2 was expressed intensely in isolated hematopoietic cells, lymphatic vessels and moderately in blood vessels. Prox-1 was absent from endothelial progenitor cells prior to lymphatic recruitment. Although vascular endothelial growth factor-165 (VEGF(sub 165)) is a key regulator of numerous cellular processes in hemangiogenesis and vasculogenesis, the role of VEGF(sub 165) in lymphangiogenesis is less clear. Exogenous VEGF(sub 165) increased blood vessel density without changing endogenous modes of vascular/lymphatic vessel formation or marker expression patterns. However, VEGF(sub 165) did increase the frequency of blood vascular anastomoses and strongly induced the antimaturational dissociation of lymphatics from blood vessels, with frequent formation of homogeneous lymphatic networks.

Parsons-Wingerter, Patricia↗

Benzo[$\mathcal{a}$]pyrene toxicokinetics in humans following dietary supplementation with 3,3'-diindolylmethane (DIM) or Brussels sprouts

Utilizing the atto-zeptomole sensitivity of UPLC-accelerator mass spectrometry (UPLC-AMS), we previously demonstrated significant first-pass metabolism following escalating (25–250 ng) oral micro-dosing in humans of [ 14 C]-benzo[a]pyrene ([ 14 C]-BaP). The present study examines the potential for supplementation with Brussels sprouts (BS) or 3,3' -diindolylmethane (DIM) to alter plasma levels of [ 14 C]-BaP and metabolites over a 48-h period following micro-dosing with 50 ng (5.4 nCi) [ 14 C]-BaP. Volunteers were dosed with [ 14 C]-BaP following fourteen days on a cruciferous vegetable restricted diet, or the same diet supplemented for seven days with 50 g of BS or 300 mg of BR-DIM ® prior to dosing. BS or DIM reduced total [ 14 C] recovered from plasma by 56–67% relative to nonintervention. Dietary supplementation with DIM markedly increased T max and reduced C max for [ 14 C]-BaP indicative of slower absorption. Further, both dietary treatments significantly reduced C max values of four downstream BaP metabolites, consistent with delaying BaP absorption. Dietary treatments also appeared to reduce the T 1/2 and the plasma AUC( 0,$\infty$ ) for Unknown Metabolite C, indicating some effect in accelerating clearance of this metabolite. Toxicokinetic constants for other metabolites followed the pattern for [ 14 C]-BaP (metabolite profiles remained relatively consistent) and non-compartmental analysis did not indicate other significant alterations. Significant amounts of metabolites in plasma were at the bay region of [ 14 C]-BaP irrespective of treatment. Although the number of subjects and large interindividual variation are limitations of this study, it represents the first human trial showing dietary intervention altering toxicokinetics of a defined dose of a known human carcinogen.

3,3'-diindolylmethane↗

HMGA2 contributes to vascular development and sprouting angiogenesis by promoting IGFBP2 production

Highlights: • Our findings demonstrates that HMGA2 promotes central arteries formation in zebrafish. • HMGA2 induces vascular development through modulation of IGFBP2 whatever in vivo or in vitro. • The mRNA expressions of HMGA2 and IGFBP2 increased significantly in peripheral blood of Hemangioma patients. • HMGA2-IGFBP2 are promising candidates towards therapeutical vascularization in vascular diseases like Hemangioma. Angiogenesis is the process by which new blood vessels form from preexisting vessels and regulates the processes of embryonic development, wound healing and tumorigenesis. HMGA2 is involved in the occurrence of several cancers, but its biological role and the exact downstream genes involved in vascular development and sprouting angiogenesis remain largely unknown. Here, we first found that HMGA2 knockdown in zebrafish embryos resulted in defects of central artery formation. RNA sequencing revealed that IGFBP2 was significantly downregulated by interference with HMGA2, and IGFBP2 overexpression reversed the inhibition of brain vascular development caused by HMGA2 deficiency. In vitro, we further found that HMGA2 knockdown blocked the migration, tube formation and branching of HUVECs. Similarly, IGFBP2 protein overexpression attenuated the impairments induced by HMGA2 deficiency. Moreover, the promotion of angiogenesis by HMGA2 overexpression was verified in a Matrigel plug assay. We next found that HMGA2 bound directly to a region in the IGFBP2 promoter and positively regulated IGFBP2 expression. Interestingly, the mRNA expression levels of HMGA2 and IGFBP2 were increased significantly in the peripheral blood of hemangioma patients, indicating that overexpression of HMGA2 and IGFBP2 results in vessel formation, consistent with the results of the in vivo and in vitro experiments. In summary, our findings demonstrate that HMGA2 promotes central artery formation by modulating angiogenesis via IGFBP2 induction.

60 APPLIED LIFE SCIENCES↗

SpRoUTS (Space Robot Universal Truss System): Reversible Robotic Assembly of Deployable Truss Structures of Reconfigurable Length

Automatic deployment of structures has been a focus of much academic and industrial work on infrastructure applications and robotics in general. This paper presents a robotic truss assembler designed for space applications - the Space Robot Universal Truss System (SpRoUTS) - that reversibly assembles a truss from a feedstock of hinged andflat-packed components, by folding the sides of each component up and locking onto the assembled structure. We describe the design and implementation of the robot and show that the assembled truss compares favorably with prior truss deployment systems.

Deployable↗

Students Planting Research Of Use To Space (SPROUTS) Networking and Engagement of Interns at Kennedy Space Center

While Kennedy Space Center (KSC) has had summer interns in the life sciences for many years, beginning in 2016, the internship program was re-envisioned. Rather than having many interns in the summer with numerous innovative small projects created primarily for this purpose, the current model is to offer internship opportunities every semester (Spring, Summer, and Fall), so that research is continuous and sustained. Since 2016 we have had 64 internship opportunities for undergraduate and graduate students in the areas of space crop production and microgravity simulation support. Some students have participated in more than one opportunity, so 52 separate individuals have participated in this program. While interns primarily worked with individual mentors on a main project, most also worked on group projects with other interns or developed smaller secondary projects on their own initiative. In addition, all interns participated in a weekly Lunch, Learn, and Discuss seminar series where they were encouraged to present their research, learn from scientists at KSC, and engage with members of the space life sciences community. When the face-to-face spring internship of 2020 ended prematurely, we resumed the weekly seminar series as a virtual event, and this led to the concept of engaging former interns. Current and former interns were invited to participate in the on-going virtual series, and 46 asked to be included on the mailing list. Our current seminar series involves remote seminars or hosted discussions from contacts within and outside of the agency, and engages a large network of current scientists and former interns with an weekly participation averaging 40-50 people. Speakers include experts from industry, academia, and other space agencies working on topics of interest to KSC space life science researchers. In addition, this series has served as a brain trust and brainstorming mechanism for KSC scientists to invite and discuss new ideas and concepts. The advent of remote work and virtual platforms has allowed us to engage with interns in a new way that will continue as future interns are integrated into this network of individuals interested in space research. This work is funded by NASA’s Space Biology Program.

Internships↗

Students Planting Research Of Use To Space (SPROUTS) Networking and Engagement of Interns at Kennedy Space Center

Beginning in 2016, the Kennedy Space Center (KSC) internship program for life sciences was re-envisioned. Rather than hosting interns in the summer only with numerous innovative small projects, the current model is to offer internship opportunities every semester, so that research is continuous and sustained. Since 2016 we have had 64 internship opportunities for undergraduate and graduate students in space crop production and microgravity simulation. 52 separate individuals have participated in this program, with some participating in more than one term. While interns primarily worked with individual mentors on a main project, most also worked on group projects or developed smaller secondary projects on their own initiative. Additionally, interns participated in a weekly Lunch, Learn, and Discuss seminar series where they presented their research, learned from scientists at KSC, and engaged with members of the space life sciences community. When the face-to-face spring 2020 internship ended prematurely, the weekly seminar series continued as a virtual event, which led to the concept of engaging former interns. Current and former interns were invited to participate in the on-going virtual series, and 46 asked to be included on the mailing list. Our current seminar series involves remote seminars from contacts within and outside of the agency, and engages a large network of current scientists and former interns with weekly participation averaging ~40 people. Speakers include experts from industry, academia, and other space agencies working on topics of interest to KSC researchers. This series has also served as a brainstorming platform for KSC scientists to discuss new ideas and concepts and invite feedback. Remote work and virtual platforms have allowed engagement in a new way that will continue as future interns are integrated into this network of individuals interested in space research. This work is funded by NASA’s Space Biology Program.

Space Crop Production↗

Rain-induced spring wheat harvest losses

When rain or a combination of rain and high humidity delay wheat harvest, losses can occur in grain yield and/or grain quality. Yield losses can result from shattering, from reduction in test weight, and in the case of windrowed grain, from rooting of sprouting grain at the soil: windrow contact. Losses in grain quality can result from reduction in test weight and from sprouting. Sprouting causes a degradation of grain proteins and starches, hence flour quality is reduced, and the grain price deteriorates to the value of feed grain. Although losses in grain yield and quality are rain-induced, these losses do not necessarily occur because a standing or windrowed crop is wetted by rain. Spike water concentration in hard red spring wheat must be increased to about 45-49% before sprouting is initiated in grain that has overcome dormancy. The time required to overcome this dormancy after the cultivar has dried to 12 to 14% water concentration differs with hard red spring cultivars. The effect of rain on threshing-ready standing and windrowed hard red spring wheat grain yeild and quality was evaluated. A goal was to develop the capability to forecast the extent of expected loss of grain yield and quality from specific climatic events that delay threshing.

Bauer, A.↗

Perturbation of tonoplast sucrose transport alters carbohydrate utilization for seasonal growth and defense metabolism in coppiced poplar

Abstract Nonstructural carbohydrate reserves of stems and roots underpin overall tree fitness and productivity under short-rotation management practices such as coppicing for bioenergy. While sucrose and starch comprise the predominant stem carbohydrate reserves of Populus, utilization for fitness and agricultural productivity is understood primarily in terms of starch turnover. The tonoplast sucrose transport protein SUT4 modulates sucrose export from source leaves to distant sinks during photoautotrophic growth, but the possibility of its involvement in remobilizing carbohydrates from storage organs during heterotrophic growth has not been explored. Here, we used PtaSUT4-knockout mutants of Populus tremula × P. alba (INRA 717-1B4) in winter (cool) and summer (warm) glasshouse coppicing experiments to assess SUT4 involvement in reserve utilization. Conditions preceding and supporting summer sprouting were considered favorable for growth, while those preceding and supporting cool temperature sprouting were suboptimal akin to conditions associated with coppicing as generally practiced. Epicormic bud emergence was delayed in sut4 mutants following lower temperature ‘winter’ but not summer coppicing. Winter xylem hexose increases were observed in control but not in sut4 stumps after coppicing. The magnitude of starch and sucrose reserve depletion was similar in control and sut4 stumps during the winter and did not explain the sprouting and xylem hexose differences. However, winter maintenance costs appeared higher in sut4 based partly on Krebs cycle intermediate levels. In control plants, bark accrual of abundant defense metabolites, including salicinoids and condensed tannins, was higher in summer than in winter, but this increase of summer defense allocations was attenuated in sut4 mutants. Temperature-sensitive trade-offs between growth and other priorities may therefore depend on SUT4 in Populus.

59 BASIC BIOLOGICAL SCIENCES↗

VEGF165 Stimulates Vessel Density and Vessel Diameter Differently in Angiogenesis and Lymphangiogenesis

Vascular endothelial growth factor-165 (VEGF(sub 165)) stimulated angiogenesis in the quail chorioallantoic membrane (CAM) by vessel expansion from the capillary network. However, lymphangiogenesis was stimulated by the filopodial guidance of tip cells located on blind-ended lymphatic sprouts. As quantified by fractal/generational branching analysis using the computer code VESGEN, vascular density increased maximally at low VEGF concentrations, and vascular diameter increased most at high VEGF concentrations. Increased vascular density and diameter were statistically independent events (r(sub s), -0.06). By fluorescence immunohistochemistry of VEGF receptors VEGFR-1 and VEGFR-2, alpha smooth muscle actin ((alpha) SMA) and a vascular/lymphatic marker, VEGF(sub 165) increased the density and diameter of sprouting lymphatic vessels guided by tip cells (accompanied by the dissociation of lymphatics from blood vessels). Isolated migratory cells expressing (alpha)SMA were recruited to blood vessels, whereas isolated cells expressing VEGFR-2 were recruited primarily to lymphatics. In conclusion, VEGF(sub 165) increased lymphatic vessel density by lymphatic sprouting, but increased blood vessel density by vascular expansion from the capillary network.

Parsons-Wingerter, Patricia↗

Occurrence of low molecular weight thiols in biological systems

Bromobimane labeling and high performance chromatography analysis were applied to various species of bacteria, plant tissues, and animal tissues. The reaction between thiols and monobromobimane is studied. Chromatograms revealing peaks produced by nonthiols and thiols are analyzed and compared. It is observed that all the bacteria species contain hydrogen sulfide, and glutathione is contained in facultative and aerobic gram-negative bacteria. For the plant tissues, the data reveal that mung bean sprouts contain homoglutathione and no glutathione; alfalfa sprouts contain homoglutathione and glutathione; the pea seed, nonlegumes, and fungi contain glutathione and no homoglutathione. It is detected that the main thiol in the animal tissues is glutathione. Based on the data, it is suggested that glutathione has an essential function in higher organisms.

Fahey, Robert C.↗

Protecting Neural Structures and Cognitive Function During Prolonged Space Flight by Targeting the Brain Derived Neurotrophic Factor Molecular Network

Brain derived neurotrophic factor (BDNF) is the main activity-dependent neurotrophin in the human nervous system. BDNF is implicated in production of new neurons from dentate gyrus stem cells (hippocampal neurogenesis), synapse formation, sprouting of new axons, growth of new axons, sprouting of new dendrites, and neuron survival. Alterations in the amount or activity of BDNF can produce significant detrimental changes to cortical function and synaptic transmission in the human brain. This can result in glial and neuronal dysfunction, which may contribute to a range of clinical conditions, spanning a number of learning, behavioral, and neurological disorders. There is an extensive body of work surrounding the BDNF molecular network, including BDNF gene polymorphisms, methylated BDNF gene promoters, multiple gene transcripts, varied BDNF functional proteins, and different BDNF receptors (whose activation differentially drive the neuron to neurogenesis or apoptosis). BDNF is also closely linked to mitochondrial biogenesis through PGC-1alpha, which can influence brain and muscle metabolic efficiency. BDNF AS A HUMAN SPACE FLIGHT COUNTERMEASURE TARGET Earth-based studies reveal that BDNF is negatively impacted by many of the conditions encountered in the space environment, including oxidative stress, radiation, psychological stressors, sleep deprivation, and many others. A growing body of work suggests that the BDNF network is responsive to a range of diet, nutrition, exercise, drug, and other types of influences. This section explores the BDNF network in the context of 1) protecting the brain and nervous system in the space environment, 2) optimizing neurobehavioral performance in space, and 3) reducing the residual effects of space flight on the nervous system on return to Earth

Schmidt, M. A.↗

An efficient virus–induced gene silencing (VIGS) system for gene functional studies in Miscanthus

Virus-induced gene silencing (VIGS) is a powerful tool for transient gene functional analysis in plants, especially for monocot species (e.g., grasses) that are recalcitrant to transformation. Despite various VIGS systems that have been developed in different plant species, none was previously available for the bioenergy crop Miscanthus. Here, we report the establishment of an efficient and robust VIGS system mediated by Tobacco Rattle Virus (TRV) in Miscanthus. We first investigated the impact of various factors that may affect gene silencing efficiency using the Miscanthus sinensis Phytoene Desaturase (MsPDS) gene as a visual indicator of photobleaching. Then, we optimized the TRV-elicited VIGS procedure using an orthogonal experimental design with four factors (sprout size, Agrobacterium concentration, vacuum infiltration time, and co-incubation time) each at three levels. The following led to the highest silencing efficiency (~76%): inoculation of germinating seedlings (1.0–2.0 mm), Agrobacterium tumefaciens culture grown to optical density at 600 nm (OD 600 ) of 0.4, vacuum infiltration for 90 min, and co-incubation for 5 h. The VIGS system established was applicable for both M. sinensis and M. lutarioriparius, with comparable gene silencing efficiency. We verified the efficacy of the VIGS system via the functional characterization of the role of a MYB transcription factor, MsMYB112, in salt stress tolerance. Expression of MsMYB112 was successfully knocked down using the VIGS system, and this led to compromised salt tolerance in the silenced Miscanthus plants. The TRV-based VIGS system established may, therefore, substantially facilitate functional genomic studies in Miscanthus.

60 APPLIED LIFE SCIENCES↗