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At least 829 records · Page 46
All sourcesNASA NTRSDOE OSTI
DOE OSTI2019

Burn and thoracic trauma alters fracture healing, systemic inflammation, and leukocyte kinetics in a rat model of polytrauma

Singular traumatic insults, such as bone fracture, typically initiate an appropriate immune response necessary to restore the host to pre-insult homeostasis with limited damage to self. However, multiple concurrent insults, such as a combination of fracture, blunt force trauma, and burns (polytrauma), are clinically perceived to result in abnormal immune response leading to inadequate healing and resolution. To investigate this phenomenon, we created a model rat model of polytrauma. To investigate relationship between polytrauma and delayed healing, we created a novel model of polytrauma in a rat which encompassed a 3-mm osteotomy, blunt chest trauma, and full-thickness scald burn. Healing outcomes were determined at 5 weeks where the degree of bone formation at the osteotomy site of polytrauma animals was compared to osteotomy only animals (OST). We observed significant differences in the bone volume fraction between polytrauma and OST animals indicating that polytrauma negatively effects wound healing. Polytrauma animals also displayed a significant decrease in their ability to return to pre-injury weight compared to osteotomy animals. Polytrauma animals also exhibited significantly altered gene expression in osteogenic pathways as well as the innate and adaptive immune response. Perturbed inflammation was observed in the polytrauma group compared to the osteotomy group as evidenced by significantly altered white blood cell (WBC) profiles and significantly elevated plasma high-mobility group box 1 protein (HMGB1) at 6 and 24 h post-trauma. Conversely, polytrauma animals exhibited significantly lower concentrations of plasma TNF-alpha (TNF-α) and interleukin 6 (IL-6) at 72 h post-injury compared to OST. Following polytrauma with burn injury, the local and systemic immune response is divergent from the immune response following a less severe singular injury (osteotomy). This altered immune response that follows was associated with a reduced capacity for wound healing.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2016

Rumen metagenome and metatranscriptome analyses of low methane yield sheep reveals a Sharpea -enriched microbiome characterised by lactic acid formation and utilisation

Enteric fermentation by farmed ruminant animals is a major source of methane and constitutes the second largest anthropogenic contributor to global warming. Reducing methane emissions from ruminants is needed to ensure sustainable animal production in the future. Methane yield varies naturally in sheep and is a heritable trait that can be used to select animals that yield less methane per unit of feed eaten. We previously demonstrated elevated expression of hydrogenotrophic methanogenesis pathway genes of methanogenic archaea in the rumens of high methane yield (HMY) sheep compared to their low methane yield (LMY) counterparts. Methane production in the rumen is strongly connected to microbial hydrogen production through fermentation processes. In this study, we investigate the contribution that rumen bacteria make to methane yield phenotypes in sheep. Using deep sequence metagenome and metatranscriptome datasets in combination with 16S rRNA gene amplicon sequencing from HMY and LMY sheep, we show enrichment of lactate-producing Sharpea spp. in LMY sheep bacterial communities. Increased gene and transcript abundances for sugar import and utilisation and production of lactate, propionate and butyrate were also observed in LMY animals. Sharpea azabuensis and Megasphaera spp. act as important drivers of lactate production and utilisation according to phylogenetic analysis and read mappings. Our findings show that the rumen microbiome in LMY animals supports a rapid heterofermentative growth, leading to lactate production. We postulate that lactate is subsequently metabolised mainly to butyrate in LMY animals, producing 2 mol of hydrogen and 0.5 mol of methane per mol hexose, which represents 24 % less than the 0.66 mol of methane formed from the 2.66 mol of hydrogen produced if hexose fermentation was directly to acetate and butyrate. These findings are consistent with the theory that a smaller rumen size with a higher turnover rate, where rapid heterofermentative growth would be an advantage, results in lower hydrogen production and lower methane formation. Together with previous methanogen gene expression data, this builds a strong concept of how animal traits and microbial communities shape the methane phenotype in sheep.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2019

Genetic and antigenic variation of foot-and-mouth disease virus during persistent infection in naturally infected cattle and Asian buffalo in India

The role of foot-and-mouth disease virus (FMDV) persistently infected ruminants in initiating new outbreaks remains controversial, and the perceived threat posed by such animals hinders international trade in FMD-endemic countries. In this study we report longitudinal analyses of genetic and antigenic variations of FMDV serotype O/ME-SA/Ind2001d sublineage during naturally occurring, persistent infection in cattle and buffalo at an organised dairy farm in India. The proportion of animals from which FMDV RNA was recovered was not significantly different between convalescent (post-clinical) and sub-clinically infected animals or between cattle and buffalo across the sampling period. However, infectious virus was isolated from a higher proportion of buffalo samples and for a longer duration compared to cattle. Analysis of the P1 sequences from recovered viruses indicated fixation of mutations at the rate of 1.816 x 10 -2 substitution/site/year (s/s/y) (95% CI 1.362–2.31 x 10 -2 s/s/y). However, the majority of point mutations were transitional substitutions. Within individual animals, the mean dN/dS (ω) value for the P1 region varied from 0.076 to 0.357, suggesting the selection pressure acting on viral genomes differed substantially across individual animals. Statistical parsimony analysis indicated that all of the virus isolates from carrier animals originated from the outbreak virus. The antigenic relationship value as determined by 2D-VNT assay revealed fluctuation of antigenic variants within and between carrier animals during the carrier state which suggested that some carrier viruses had diverged substantially from the protection provided by the vaccine strain. This study contributes to understanding the extent of within-host and within-herd evolution that occurs during the carrier state of FMDV.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2019

Structures of TOG1 and TOG2 from the human microtubule dynamics regulator CLASP1

Tubulin-binding TOG domains are found arrayed in a number of proteins that regulate microtubule dynamics. While much is known about the structure and function of TOG domains from the XMAP215 microtubule polymerase family, less in known about the TOG domain array found in animal CLASP family members. The animal CLASP TOG array promotes microtubule pause, potentiates rescue, and limits catastrophe. How structurally distinct the TOG domains of animal CLASP are from one another, from XMAP215 family TOG domains, and whether a specific order of structurally distinct TOG domains in the TOG array is conserved across animal CLASP family members is poorly understood. We present the x-ray crystal structures of Homo sapiens (H.s.) CLASP1 TOG1 and TOG2. The structures of H.s. CLASP1 TOG1 and TOG2 are distinct from each other and from the previously determined structure of Mus musculus (M.m.) CLASP2 TOG3. Comparative analyses of CLASP family TOG domain structures determined to date across species and paralogs supports a conserved CLASP TOG array paradigm in which structurally distinct TOG domains are arrayed in a specific order. H.s. CLASP1 TOG1 bears structural similarity to the free-tubulin binding TOG domains of the XMAP215 family but lacks many of the key tubulin-binding determinants found in XMAP215 family TOG domains. This aligns with studies that report that animal CLASP family TOG1 domains cannot bind free tubulin or microtubules. In contrast, animal CLASP family TOG2 and TOG3 domains have reported microtubule-binding activity but are structurally distinct from the free-tubulin binding TOG domains of the XMAP215 family. H.s. CLASP1 TOG2 has a convex architecture, predicted to engage a hyper-curved tubulin state that may underlie its ability to limit microtubule catastrophe and promote rescue. M.m. CLASP2 TOG3 has unique structural elements in the C-terminal half of its α-solenoid domain that our modeling studies implicate in binding to laterally-associated tubulin subunits in the microtubule lattice in a mode similar to, yet distinct from those predicted for the XMAP215 family TOG4 domain. The potential ability of the animal CLASP family TOG3 domain to engage lateral tubulin subunits may underlie the microtubule rescue activity ascribed to the domain. These findings highlight the structural diversity of TOG domains within the CLASP family TOG array and provide a molecular foundation for understanding CLASP-dependent effects on microtubule dynamics.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2014

Missouri Agricultural Energy Saving Team-A Revolutionary Opportunity (MAESTRO)

The Missouri Agricultural Energy Saving Team-A Revolutionary Opportunity (MAESTRO) program brought together a team of representatives from government, academia, and private industry to enhance the availability of energy efficiency services for small livestock producers in the State of Missouri. The Missouri Department of Agriculture (MDA) managed the project via a subcontract with the University of Missouri (MU), College of Agriculture Food and Natural Resources, MU Extension, the MU College of Human Environmental Sciences, the MU College of Engineering, and the Missouri Agricultural and Small Business Development Authority (MASBDA). MU teamed with EnSave, Inc, a nationally-recognized expert in agricultural energy efficiency to assist with marketing, outreach, provision of farm energy audits and customer service. MU also teamed with independent home contractors to facilitate energy audits of the farm buildings and homes of these livestock producers. The goals of the project were to: (1) improve the environment by reducing fossil fuel emissions and reducing the total energy used on small animal farms; (2) stimulate the economy of local and regional communities by creating or retaining jobs; and (3) improve the profitability of Missouri livestock producers by reducing their energy expenditures. Historically, Missouri scientists/engineers conducted programs on energy use in agriculture, such as in equipment, grain handling and tillage practices. The MAESTRO program was the first to focus strictly on energy efficiency associated with livestock production systems in Missouri and to investigate the applicability and potential of addressing energy efficiency in animal production from a building efficiency perspective. A. Project Objectives The goal of the MAESTRO program was to strengthen the financial viability and environmental soundness of Missouri's small animal farms by helping them implement energy efficient technologies for the production facility, farm buildings, and the homes on these farms. The expected measurable outcomes of the project were to improve the environment and stimulate the economy by: • Reducing annual fossil fuel emissions by 1,942 metric tons of carbon dioxide equivalent, reducing the total annual energy use on at least 323 small animal farms and 100 farm homes by at least 8,000 kWh and 2,343 therms per farm. • Stimulating the economy by creating or retaining at least 69 jobs, and saving small animal farmers an average of $2,071 per farm in annual energy expenditures. B. Project Scope The MAESTRO team chose the target population of small farms because while all agriculture is traditionally underserved in energy efficiency programs, small farms were particularly underserved because they lack the financial resources and access to energy efficiency technologies that larger farms deploy. The MAESTRO team reasoned that energy conservation, financial and educational programs developed while serving the agricultural community could serve as a national model for other states and their agricultural sectors. The target population was approximately 2,365 small animal farm operations in Missouri, specifically those farms that were not by definition a confined animal feeding operation (CAFO). The program was designed to create jobs by training Missouri contractors and Missouri University Extension staff how to conduct farm audits. The local economy would be stimulated by an increase in construction activity and an increasing demand for energy efficient farm equipment. Additionally, the energy savings were deemed critical in keeping Missouri farms in business. This project leveraged funds using a combination of funds from the Missouri Department of Natural Resources’ Missouri Energy Center and its Soil and Water Conservation Program, from the state's Linked Deposits, MASBDA's agricultural loan guarantee programs, and through the in-kind contribution of faculty and staff time to the project from these agencies and MU. Several hundred Missouri livestock producers were contacted during the MAESTRO project. Of the livestock producers, 254 invited the team to conduct a farm energy assessment which complied with ASABE 612. A total of 147 livestock farm upgrades were implemented, representing 57.5 percent of the farms for which a farm energy assessment was completed. This represented a statewide average annual savings of 1,088,324 kWh and 75,516 therms. The team also reviewed the condition of the livestock producer’s home(s). A total of 106 home energy assessments were completed and 48 individual homes implemented their recommended upgrades, representing 45 percent of the farm homes for which an energy assessment was completed. This represented a statewide average annual savings of 323,029 kWh, and 769.4 therms. More of these farmers likely would have updated their homes but the funding to incentivize them fell short. In spite of the shortfall in incentive funds, some farmers still updated their homes as they saw the value in making these changes to their home.

32 ENERGY CONSERVATION, CONSUMPTION, AND UTILIZATI↗
DOE OSTI2026

Producing cultivated meat cuts that match traditional meat in quality and affordability (CRADA Final Report)

Novel Farms, Inc., as a participant of the Cyclotron Road Lab-Embedded Entrepreneurship Program, investigated the performance of various animal cell lines and animal cell media formulations using its proprietary scaffolds as a growth substratum towards the development of economically-feasible cultivated meat production. As an alternative to conventionally farmed animal meat, cultivated meat production has the potential to yield a marginal carbon footprint while providing sustainable and cruelty-free animal protein. The prohibitively high cost of the growth factors necessary in animal cell culture media constitutes the primary barrier that hinders the economically viable production of cultivated meat products. Furthermore, scaffolding is required to create structured meat products, but current options are unsuitable for cost-effective large-scale production of cultivated meat. Novel Farms has developed a proprietary technology to produce scaffolding that has the potential to reduce production costs while providing an ideal 3D surface for cell growth. Developing cost-efficient methods for cultivated meat production will be vital in satisfying the growing demand for animal protein while avoiding the deforestation, carbon emissions, biodiversity loss and other negative outcomes of industrial animal farming.

59 BASIC BIOLOGICAL SCIENCES↗
NASA NTRS1971

Erythropoietic radiosensitivity of the rat during altitude acclimatization.

The effect of a sublethal dose (300 R) of X-radiation upon the erythropoietic system of the rat, during 60-day acclimatization to moderate hypoxia (3,800 m altitude), was studied. Past work has shown that hypoxic animals are damaged less by radiation than animals irradiated in a normal environment; therefore, it was postulated that if, after acclimatization to hypoxia the bone marrow oxygen tension returns to sea-level values, these animals should suffer radiation damage equivalent to animals at sea level. The principal parameters followed were the rate of depletion of injected Fe-59 from the plasma of chronically catheterized rats, and its subsequent reappearance in the circulating erythrocytes. After 20 days of acclimatization, both parameters for altitude-irradiated animals returned to the values of animals irradiated at sea level, previously having reflected increased erythropoiesis. In altitude nonirradiated animals the parameters indicated erythropoietic stimulation persisting up to 45 days acclimatization. The protective effect of the hypoxia on the stem cells vanished during acclimatization, presumably as cellular oxygen tension rose.

Gaugl, J. F.↗
NASA NTRS1977

Veterinary medical considerations for the use of nonhuman primates in space research

The validity of biomedical research using animal subjects is highly dependent on the use of 'normal' and healthy animals. The current costs of research programs dictate that a minimum number of animals and test replicates be used to obtain the desired data. The use of healthy and standardized animals increases the probability of obtaining valid data while also permitting greater economy by reducing the between-individual variation, thus allowing the use of fewer animals. Areas of concern when planning animal payloads include constraints of the flight on candidate species selection, screening for physiological and psychological normalcy, procedures for routine care and quarantine of new animals and those returning from space, ground-based studies to determine experimental protocol, selection of instrumentation, stress during transportation for flight operations, housing and care facilities at launch and recovery sites, and the overall veterinary program.

Simmonds, R. C.↗
NASA NTRS1993

Evaluation of medical treatments to increase survival of ebullism in guinea pigs

Spaceflight carriers run a constant risk of exposure to vacuum. Above 63,000 ft (47 mmHg), the ambient pressure falls below the vapor pressure of water at 37 C, and tissue vaporization (ebullism) begins. Little is know about appropriate resuscitative protocols after such an ebullism exposure. This study identified injury patterns and mortality rates associated with ebullism while verifying effectiveness of traditional pulmonary resuscitative techniques. Male Hartley guinea pigs were exposed to 87,000 ft for periods of 40 to 115 sec. After descent, those animals that did not breathe spontaneously were given artificial ventilation by bag and mask for up to 15 minutes. Those animals surviving were randomly assigned to one of three treatment groups--hyperbaric oxygen (HBO), ground-level oxygen (GLO2), and ground-level air (GLAIR). The HBO group was treated on a standard treatment table 6A while the GLO2 animals received O2 for an equivalent length of time. Those animals in the GLAIR group were observed only. All surviving animals were humanely sacrified at 48 hours. Inflation of the animal's lungs after the exposure was found to be difficult and, at times, impossible. This may be due to surfactant disruption at the alveolar lining. Electron microscopy identified a disruption of the surfactant layer in animals that did not survive initial exposure. Mortality was found to increase with exposure time: 40 sec--0 percent; 60 sec--6 percent; 70 sec--40 percent; 80 sec--13 percent; 100 sec--38 percent; 110 sec--40 percent; and 115 sec--100 percent. There was no difference in the delayed mortality among the treatment groups (HBO--15 percent, GLO2--11 percent, GLAIR--11 percent). However, since resuscitation was ineffective, the effectiveness of any post-exposure treatment was severely limited. Preliminary results indicate that reuscitation of guinea pigs following ebullism exposure is difficult, and that current techniques (such as traditional CPR) may not be appropriate.

Stegmann, Barbara J.↗
NASA NTRS1997

BP and Vascular Function Following Space Flight

Blood pressure and mesenteric resistance artery function were assessed in 9-week-old spontaneously hypertensive rats following an 18 day shuttle flight on STS-80. Blood pressure was measured twice, first in conscious animals using a tail-cuff method and then while the animals were anesthetized with 2% halothane in O2. Isolated mesenteric resistance artery responses to cumulative additions of norepinephrine, acetylcholine, sodium nitroprusside, and calcium were measured within 17 hours of landing using wire myography. Blood pressure was slightly reduced in conscious animals following flight (p=0.056) but was significantly elevated (p less than.001) above vivarium control group values in anesthetized animals. Maximal contraction of mesenteric arteries to norepinephrine was attenuated in the flight animals (p less than.001)aswasrelaxationtoacetylcholine(p less than .001)andcalcium(p less than .05). There was no difference between flight and control animals in the vessel response to sodium nitroprusside (p greater than .05). The results suggest that there may have been an increase in synthesis and release of nitric oxide in the flight animals.

Hatton, Daniel C.↗
DOE OSTI2017

Local and systemic changes associated with long-term, percutaneous, static implantation with titanium alloys in rhesus macaques (Macaca mulatta)

Metal alloys are frequently used as implant materials in veterinary medicine. Recent studies suggest that many types of metal alloys may induce both local and systemic inflammatory responses. In this study, 37 rhesus macaques with long-term skull-anchored percutaneous titanium alloy implants (0-14 years duration) were evaluated for changes in their hematology, coagulation and serum chemistry profiles. Negative controls (n=28) did not have implants. All of the implanted animals were on IACUC-approved protocols and were not implanted for the purpose of this study. Animals with implants had significantly higher plasma D-dimer and lower antithrombin III concentrations compared with nonimplanted animals (p-values < 0.05). Additionally, animals with implants had significantly higher globulin, and lower albumin and calcium concentrations compared with nonimplanted animals (p-values < 0.05). Many of these changes were positively correlated with duration of implantation as well as the number of implants. Chronic bacterial infection was observed on the skin around many of the implant sites, and within deeper tissues. Representative histopathology around the implant site of two implanted animals revealed chronic suppurative to pyogranulomatous inflammation extending from the skin to the dura mater. X-ray fluorescence microscopy of tissue biopsies from the implant site of the same two animals revealed significant increases in free metal ions within the tissue, including titanium and iron. Free metal ions persisted in the tissues up to 6 months postexplant. These results suggest that long-term skull-anchored percutaneous titanium alloy implants results in localized inflammation, chronic infection, and leaching of metal ions into local tissues.

Frydman, Galit F.↗
DOE OSTI2015

SU-C-303-04: Evaluation of On- and Off-Line Bioluminescence Tomography System for Focal Irradiation Guidance

Purpose: We have developed offline and on-board bioluminescence tomography(BLT) systems for the small animal radiation research platform(SARRP) for radiation guidance of soft tissue targets. We investigated the effectiveness of offline BLT guidance. Methods: CBCT is equipped on both the offline BLT system and SARRP that are 10 ft. apart. To evaluate the setup error during animal transport between the two systems, we implanted a luminescence source in the abdomen of anesthetized mice. Five mice were studied. After CBCT was acquired on both systems, source centers and correlation coefficients were calculated. CBCT was also used to generate object mesh for BLT reconstruction. To assess target localization, we compared the localization of the luminescence source based on (1)on-board SARRP BLT and CBCT, (2)offline BLT and CBCT, and (3)offline BLT and SARRP CBCT. The 3rd comparison examines if an offline BLT system can be used to guide radiation when there is minimal target contrast in CBCT. Results: Our CBCT results show the offset of the light source center can be maintained within 0.2 mm during animal transport. The center of mass(CoM) of the light source reconstructed by the offline BLT has an offset of 1.0 ± 0.4 mm from the ‘true’ CoM as derived from the SARRP CBCT. The results compare well with the offset of 1.0 ± 0.2 mm using on-line BLT. Conclusion: With CBCT information provided by the SARRP and effective animal immobilization during transport, these findings support the use of offline BLT in close vicinity for accurate soft tissue target localization for irradiation. However, the disadvantage of the off-line system is reduced efficiency as care is required to maintain stable animal transport. We envisage a dual use system where the on-board arrangement allows convenient access to CBCT and avoids disturbance of animal setup. The off-line capability would support standalone longitudinal imaging studies. The work is supported by NIH R01CA158100 and Xstrahl Ltd. Drs. John Wong and Iulian Iordachita receive royalty payment from a licensing agreement between Xstrahl Ltd and Johns Hopkins University. John Wong also has a consultant agreement with Xstrahl Ltd.

60 APPLIED LIFE SCIENCES↗
NASA NTRS1996

Calbindins decreased after space flight

Exposure of the body to microgravity during space flight causes a series of well-documented changes in Ca2+ metabolism, yet the cellular and molecular mechanisms leading to these changes are poorly understood. Calbindins, vitamin D-dependent Ca2+ binding proteins, are believed to have a significant role in maintaining cellular Ca2+ homeostasis. In this study, we used biochemical and immunocytochemical approaches to analyze the expression of calbindin-D28k and calbindin-D9k in kidneys, small intestine, and pancreas of rats flown for 9 d aboard the space shuttle. The effects of microgravity on calbindins in rats from space were compared with synchronous Animal Enclosure Module controls, modeled weightlessness animals (tail suspension), and their controls. Exposure to microgravity resulted in a significant and sustained decrease in calbindin-D28k content in the kidney and calbindin-D9k in the small intestine of flight animals, as measured by enzyme-linked immunosorbent assay (ELISA). Modeled weightlessness animals exhibited a similar decrease in calbindins by ELISA. Immunocytochemistry (ICC) in combination with quantitative computer image analysis was used to measure in situ the expression of calbindins in the kidney and the small intestine, and the expression of insulin in pancreas. There was a large decrease of immunoreactivity in renal distal tubular cell-associated calbindin-D28k and in intestinal absorptive cell-associated calbindin-D9k of space flight and modeled weightlessness animals compared with matched controls. No consistent difference in pancreatic insulin immunoreactivity between space flight, modeled weightlessness, and controls was observed. Regression analysis of results obtained by quantitative ICC and ELISA for space flight, modeled weightlessness animals, and their controls demonstrated a significant correlation. These findings after a short-term exposure to microgravity or modeled weightlessness suggest that a decreased expression of calbindins may contribute to the disorders of Ca2+ metabolism induced by space flight.

Non-NASA Center↗
NASA NTRS1990

Nocturnal illumination maintains reproductive function and simulates the period-lengthening effect of constant light in the mature male Djungarian hamster (Phodopus sungorus)

Mature male Djungarian hamsters (Phodopus sungorus) were placed in individual light-tight, sound attenuated chambers and exposed to one of four lighting conditions for a duration of approximately seven weeks. The four lighting conditions were: constant light (LL); constant dark (DD); feedback lighting (LDFB; a condition that illuminates the cage in response to locomotor activity); or a feedback lighting neighbor control (LDFB NC; the animal receives the same light pattern as a paired animal in feedback lighting, but has no control over it). Exposure of hamsters to LL or LDFB produced significantly and similarly longer free-running periods of the locomotor activity rhythm than exposure of animals to DD. Hamsters exposed to LDFB NC did not free-run or entrain, but rather displayed "relative coordination". The paired testes and sex accessory glands weights suggest that in the Djungarian hamster, LL and LDFB exposed animals maintained reproductive function, whereas DD exposed animals did not. Animals exposed to LDFB NC had intermediate paired testes weights. Since several previous studies have demonstrated that short pulses of light, which are coincident with the subjective night, are photostimulatory, it is not surprising that LDFB maintained reproductive function in the mature Djungarian hamster. Feedback lighting, however, has been shown to be an insufficient stimulus to maintain reproductive function of mature male and female Syrian hamsters, and to the reproductive maturation of immature Djungarian hamsters. The results suggest that there may be slight, but significant differences in the way these two species interpret photoperiod, as well as a developmental change in the photoperiodic response of Djungarian hamsters.

NASA Program Space Biology↗
NASA NTRS1993

Deep cytoplasmic rearrangements in ventralized Xenopus embryos

Following fertilization in Xenopus, dramatic rearrangements of the egg cytoplasm relocalize maternally synthesized egg components. During the first cell cycle the vegetal yolk mass rotates relative to the egg surface, toward the sperm entry point (SEP) (J. P. Vincent, G. F. Oster, and J. C. Gerhart, 1986, Dev. Biol. 113, 484-500), while concomitant deep cytoplasmic rearrangements occur in the animal hemisphere (M. V. Danilchik and J. M. Denegre, 1991, Development 111, 845-856). In this paper we examine the role of vegetal yolk mass rotation in producing the animal cytoplasmic rearrangements. We inhibited rotation by uv-irradiating embryos during the first cell cycle, a treatment that yields an extremely ventralized phenotype. Both uv-irradiated embryos and unirradiated control embryos show cytoplasmic rearrangements in the animal hemisphere during the first cell cycle. Cytoplasmic rearrangements on the SEP side of the embryo associated with the path of the sperm pronucleus, plus a swirl on the anti-SEP (dorsal) side, are seen, whether or not yolk mass rotation has occurred. This result suggests a role for the expanding sperm aster in directing animal hemisphere cytoplasmic movements. In unirradiated control embryos the anti-SEP (dorsal) swirl is larger than that in uv-irradiated embryos and often extends into the vegetal hemisphere, consistent with the animal cytoplasm having been pulled dorsally and vegetally by the sliding vegetal yolk mass. Thus the yolk mass rotation may normally enhance the dorsalward cytoplasmic movement, begun by the sperm aster, enough to induce normal axis formation. We extended our observations of unirradiated control and uv-irradiated embryos through early cleavages. The vegetal extent of the anti-SEP (dorsal) swirl pattern seen in control embryos persists through the early cleavage period, such that labeled animal cytoplasm extends deep into dorsal third-tier blastomeres at the 32-cell stage. Significantly, in uv-irradiated embryos, which have not undergone vegetal rotation, most of this labeled material remains more equatorial.

Non-NASA Center↗
DOE OSTI2018

Age-dependent behaviors, seizure severity and neuronal damage in response to nerve agents or the organophosphate DFP in immature and adult rats

Exposure to nerve agents (NAs) and other organophosphates (OPs) can initiate seizures that rapidly progress to status epilepticus (SE). While the electrographic and neuropathological sequelae of SE evoked by NAs and OPs have been characterized in adult rodents, they have not been adequately investigated in immature animals. In this study postnatal day (PND) 14, 21 and 28 rat pups, along with PND70 animals as adult controls, were exposed to NAs (sarin, VX) or another OP (diisopropylfluorophosphate, DFP). We then evaluated behavioral and electrographic (EEG) correlates of seizure activity, and performed neuropathology using Fluoro-Jade B. Although all immature rats exhibited behaviors that are often characterized as seizures, the incidence, duration, and severity of the electrographic seizure activity were age-dependent. No (sarin and VX) or brief (DFP) EEG seizure activity was evoked in PND14 rats, while SE progressively increased in severity as a function of age in PND21, 28 and 70 animals. Fluoro-Jade B staining was observed in multiple brain regions of animals that exhibited prolonged seizure activity. Neuronal injury in PND14 animals treated with DFP was lower than in older animals and absent in rats exposed to sarin or VX. In conclusion, we found that NAs and an OP provoked robust SE and neuronal injury similar to adults in PND21 and PND28, but not in PND14, rat pups. Convulsive behaviors were often present independent of EEG seizures and were unaccompanied by neuronal damage. These differential responses should be considered when investigating medical countermeasures for NA and OP exposure in pediatric populations.

Neurosciences & Neurology↗
DOE OSTI2023

The Sustainability Index: a new tool to breed for reduced greenhouse-gas emissions intensity in Australian dairy cattle

Context The Australian dairy industry has a target to reduce greenhouse-gas (GHG) emissions intensity by 30% between 2015 and 2030. At the animal level, apart from nutritional modifications and other management practices, selecting animals that emit less GHG can be a cost-effective and long-term strategy. Given the world’s demand for protein is increasing, selecting for animals with lower GHG emissions per unit of production, i.e. emissions intensity, is a realistic approach that addresses the key issue of emissions reduction while maintaining farm productivity. Aim To develop a selection index for Australian dairy cattle to breed for reduced emissions intensity. Methods The Sustainability Index was built based on the existing Balanced Performance Index (BPI) but placed greater emphasis on production, survival, health and feed efficiency. Key results In August 2022, DataGene released the Sustainability Index that can be used by dairy farmers to select animals with lower environmental footprints. Compared with BPI, the weights for protein, fat, survival, mastitis resistance and feed efficiency increased by 2.6-, 1.4-, 2.8-, 1.3- and 3.8-fold respectively. It is expected that with the use of the Sustainability Index, emissions intensity will be reduced by 7.64%, 8.96% and 5.52% respectively in Holstein, Jersey and Red breeds by 2050, compared with the 2015 level. The corresponding values when selecting for BPI were 6.34%, 7.91% and 5.23% respectively. However, the trade-off in BPI when using the Sustainability Index will be AUD0.79, AUD0.83, AUD0.22 per cow per year for Holstein, Jersey and Red breeds respectively. Conclusions The current profit index BPI has contributed to reduction in emissions intensity. To enhance the rate of improvement in emissions intensity, the Sustainability Index can be used with minimal impacts on profit. Implications To breed for lower emissions intensity, farmers can select animals with high Sustainability Index values that are published on DataVat (datavat.com.au) and the Good Bulls App. To lower gross emissions, combinations of additional measures such as management of diet, adjustment to animal numbers, management of stored manure, and appropriate use of carbon neutral fertiliser, renewable fuels and energy, will need to be adopted on farms.

Agriculture↗
DOE OSTI2022

Deletion of E184L, a Putative DIVA Target from the Pandemic Strain of African Swine Fever Virus, Produces a Reduction in Virulence and Protection against Virulent Challenge

African swine fever (ASF) is currently causing a major pandemic affecting the swine industry and protein availability from Central Europe to East and South Asia. No commercial vaccines are available, making disease control dependent on the elimination of affected animals. Here, we show that the deletion of the African swine fever virus (ASFV) E184L gene from the highly virulent ASFV Georgia 2010 (ASFV-G) isolate produces a reduction in virus virulence during the infection in swine. Of domestic pigs intramuscularly inoculated with a recombinant virus lacking the E184L gene (ASFV-GDE184L), 40% experienced a significantly (5 days) delayed presentation of clinical disease and, overall, had a 60% rate of survival compared to animals inoculated with the virulent parental ASFV-G. Importantly, all animals surviving ASFV-G-DE184L infection developed a strong antibody response and were protected when challenged with ASFV-G. As expected, a pool of sera from ASFV-G-DE184L-inoculated animals lacked any detectable antibody response to peptides partially representing the E184L protein, while sera from animals inoculated with an efficacious vaccine candidate, ASFV-GDMGF, strongly recognize the same set of peptides. These results support the potential use of the E184L deletion for the development of vaccines able to differentiate infected from vaccinated animals (DIVA). Therefore, it is shown here that the E184L gene is a novel ASFV determinant of virulence that can potentially be used to increase safety in preexisting vaccine candidates, as well as to provide them with DIVA capabilities. To our knowledge, E184L is the first ASFV gene product experimentally shown to be a functional DIVA antigenic marker.

59 BASIC BIOLOGICAL SCIENCES↗