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At least 415 records · Page 23

Towards Construction of a Novel Nanometer-Resolution MeV-STEM for Imaging Thick Frozen Biological Samples

Driven by life-science applications, a mega-electron-volt Scanning Transmission Electron Microscope (MeV-STEM) has been proposed here to image thick frozen biological samples as a conventional Transmission Electron Microscope (TEM) may not be suitable to image samples thicker than 300–500 nm and various volume electron microscopy (EM) techniques either suffering from low resolution, or low speed. The high penetration of inelastic scattering signals of MeV electrons could make the MeV-STEM an appropriate microscope for biological samples as thick as 10 μm or more with a nanoscale resolution, considering the effect of electron energy, beam broadening, and low-dose limit on resolution. The best resolution is inversely related to the sample thickness and changes from 6 nm to 24 nm when the sample thickness increases from 1 μm to 10 μm. To achieve such a resolution in STEM, the imaging electrons must be focused on the specimen with a nm size and an mrad semi-convergence angle. This requires an electron beam emittance of a few picometers, which is ~1000 times smaller than the presently achieved nm emittance, in conjunction with less than 10 -4 energy spread and 1 nA current. We numerically simulated two different approaches that are potentially applicable to build a compact MeV-STEM instrument: (1) DC (Direct Current) gun, aperture, superconducting radio-frequency (SRF) cavities, and STEM column; (2) SRF gun, aperture, SRF cavities, and STEM column. Beam dynamic simulations show promising results, which meet the needs of an MeV-STEM, a few-picometer emittance, less than 10 -4 energy spread, and 0.1–1 nA current from both options. Also, we designed a compact STEM column based on permanent quadrupole quintuplet, not only to demagnify the beam size from 1 μm at the source point to 2 nm at the specimen but also to provide the freedom of changing the magnifications at the specimen and a scanning system to raster the electron beam across the sample with a step size of 2 nm and the repetition rate of 1 MHz. This makes it possible to build a compact MeV-STEM and use it to study thick, large-volume samples in cell biology.

36 MATERIALS SCIENCE↗

X69R Is a Non-Essential Gene That, When Deleted from African Swine Fever, Does Not Affect Virulence in Swine

African swine fever virus (ASFV) is currently causing devastating outbreaks in Asia and Europe, and the ASFV strain Georgia (ASFV-G) is responsible for these outbreaks. ASFV-G is highly virulent and continues to be maintained in these outbreak areas, apparently without suffering significant genomic or phenotypic changes. When comparing the genome of ASFV-G to other isolates, a thus-far uncharacterized gene, X69R, is highly conserved and, interestingly, is similar to another ASFV uncharacterized gene, J64R. All sequenced ASFV isolates have one or both of these genes, X69R or J64R, suggesting that the presence of at least one of these genes may be necessary for ASFV replication and or virulence. The X69R gene is present in the ASFV-G genome while J64R is absent. To assess the importance of X69R in ASFV-G functionality, we developed a recombinant virus by deleting the X69R gene from the ASFV-G genome (ASFV-G-ΔX69R). ASFV-G-ΔX69R had the same replication kinetics in primary swine macrophage cultures as the parental ASFV-G, indicating that the X69R gene is not essential for ASFV-G viability or efficient replication in the main target cell during in vivo infection. In addition, swine intramuscularly inoculated with a low dose (102 HAD50) of ASFV-G-ΔX69R developed a clinical disease indistinguishable from that induced by the same dose of the virulent parental ASFV-G isolate. Viremia values of ASFV-G-ΔX69R did not significantly differ from those detected in animals infected with parental virus. Therefore, deletion of the X69R gene from ASFV-G does not affect virus replication or virulence in swine.

59 BASIC BIOLOGICAL SCIENCES↗

African Swine Fever Vaccine Candidate ASFV-G-ΔI177L Produced in the Swine Macrophage-Derived Cell Line IPKM Remains Genetically Stable and Protective against Homologous Virulent Challenge

ASFV vaccine candidate ASFV-G-ΔI177L has been shown to be highly efficacious in inducing protection against challenges with the parental virus, the Georgia 2010 isolate, as well as against field strains isolated from Vietnam. ASFV-G-ΔI177L has been shown to produce protection even when used at low doses (10 2 HAD 50 ) and shows no residual virulence even when administered at high doses (10 6 HAD 50 ) or evaluated for a relatively long period of time (6 months). ASFV-G-ΔI177L stocks can only be massively produced in primary cell macrophages. Alternatively, its modified version (ASFV-G-ΔI177L/ΔLVR) grows in a swine-derived cell line (PIPEC), acquiring significant genomic modifications. We present here the development of ASFV-G-ΔI177L stocks in a swine macrophage cell line, IPKM, and its protective efficacy when evaluated in domestic pigs. Successive passing of ASFV-G-ΔI177L in IPKM cells produces minimal genomic changes. Interestingly, a stock of ASFV-G-ΔI177L obtained after 10 passages (ASFV-G-ΔI177Lp10) in IPKM cells showed very small genomic changes when compared with the original virus stock. ASFV-G-ΔI177Lp10 conserves similar growth kinetics in primary swine macrophage cultures than the original parental virus ASFV-G-ΔI177L. Pigs infected with 10 3 HAD 50 of ASFV-G-ΔI177Lp10 developed a strong virus-specific antibody response and were completely protected against the challenge with the parental virulent field isolate Georgia 2010. Therefore, IPKM cells could be an effective alternative for the production of ASFV vaccine stocks for those vaccine candidates exclusively growing in swine macrophages.

59 BASIC BIOLOGICAL SCIENCES↗

A Self-Assembling Pfs230D1-Ferritin Nanoparticle Vaccine Has Potent and Durable Malaria Transmission-Reducing Activity

Malaria is caused by eukaryotic protozoan parasites of the genus Plasmodium. There are 249 million new cases and 608,000 deaths annually, and new interventions are desperately needed. Malaria vaccines can be divided into three categories: liver stage, blood stage, or transmission-blocking vaccines. Transmission-blocking vaccines prevent the transmission of disease by the mosquito vector from one human to another. Pfs230 is one of the leading transmission-blocking vaccine antigens for malaria. Here, we describe the development of a 24-copy self-assembling nanoparticle vaccine comprising domain 1 of Pfs230 genetically fused to H. pylori ferritin. The single-component Pfs230D1-ferritin construct forms a stable and homogenous 24-copy nanoparticle with good production yields. The nanoparticle is highly immunogenic, as two low-dose vaccinations of New Zealand White rabbits elicited a potent and durable antibody response with high transmission-reducing activity when formulated in two distinct adjuvants suitable for translation to human use. This single-component 24-copy Pfs230D1-ferritin nanoparticle vaccine has the potential to improve production pipelines and the cost of manufacturing a potent and durable transmission-blocking vaccine for malaria control.

60 APPLIED LIFE SCIENCES↗

Further considerations of spallation effects

Trapped photon and cosmic ray effects on spallation in the UK-5 (hard X ray telescope) central crystal were measured. Both low dose and high dose effects were considered. Decay results are presented in tables.

Dyer, C.↗

Development and investigation of single-scan TV radiography for the acquisition of dynamic physiologic data

Research data obtained by the low dose electronic radiography system are reported. Data cover: (1) localization and tracking of Ta screws implanted in the inner wall of the right ventrical of the heart, (2) use of cross hairs to outline inner or outer heart wall contours, (3) quantitative measure of anatomical components which are stationary in size or change size dynamically, and (4) study of dynamic quantitative data from roentenologic or fluoroscopic procedures.

Baily, N. A.↗

Brain catechol synthesis - Control by brain tyrosine concentration

Brain catechol synthesis was estimated by measuring the rate at which brain dopa levels rose following decarboxylase inhibition. Dopa accumulation was accelerated by tyrosine administration, and decreased by treatments that lowered brain tyrosine concentrations (for example, intraperitoneal tryptophan, leucine, or parachlorophenylalanine). A low dose of phenylalanine elevated brain tyrosine without accelerating dopa synthesis. Our findings raise the possibility that nutritional and endocrine factors might influence brain catecholamine synthesis by controlling the availability of tyrosine.

Wurtman, R. J.↗

Total body calcium analysis using the Ca-12(n, alpha) Ar-37 reaction

A low dose neutron activation technique was developed to measure total body calcium in vivo. The effort had included development of irradiation and processing facilities and conduction of human studies to determine the accuracy and precision of measurement attainable with the systems.

Lewellen, T. K.↗

Application of fiber optics in spacecraft

The unique requirements placed on an optical fiber data system in space are briefly reviewed in this paper. These include temperature extremes, particle radiation, and, in some cases, a stringent electrical power budget. An overriding need is for the best possible reliability. Results related to the power consumption of a fiber terminal pair are reported, with the conclusion that a 32-port transmission star bus configuration could be operated error-free at 10 Mbits/s on 1/8 watt per transmitting terminal. The design of a transmitter terminal optimized for low idling power consumption is discussed. Although the electronic elements on a spacecraft are protected, the fiber cable may be subjected to large temperature extremes. Measured effects of temperature on fibers and cables are discussed and related to bus design. Radiation effects are briefly reviewed, with special reference to the long-duration low dose rate environment of a spacecraft.

Johnston, A. R.↗

Images of single X-ray photons from X-ray phosphor screens

Photographs show the efficiency and resolution characteristics of single X-ray photons converted to optical photons in a variety of commercial X-ray phosphor screens. The recording system uses a cooled, two-stage image intensifier system with fiber optics coupling to the phosphor screen. High efficiencies in the energy range 20-100 keV with position resolution of the order 200 microns are readily achievable, although the sensitive area is presently limited. Potential applications include X-ray astronomy, high-speed X-ray diffractometry, and extremely low dose radiography.

Fishman, G. J.↗

Drastic reduction of adsorption of CO and H2 on (111)-type Pd layers

Clean surfaces of (111)-type Pd layers, grown from the vapor phase on Mo(110) at room temperature, were used to study the adsorption of CO and H2 by temperature-programmed desorption, Auger electron spectroscopy, and low-energy electron diffraction. Mild annealing of the as-grown layers during a single desorption cycle (to about 600 K) drastically reduces the adsorption for both adsorbates. Low-dose argon-ion bombardment introduces surface imperfections which restore a high adsorption probability. The results are interpreted in terms of particular (111)-type surface structures that persist tp layer thicknesses of about four monolayers; the results raise questions with respect to the surface structure of supported thin epitaxial islands and particles of Pd and possibly also with respect to conventional methods of preparing bulk surfaces of Pd for adsorption studies.

Poppa, H.↗

Space environment effects on fiber optics systems

The performance survivability of hardened fiber optic data link design for application in future spacecraft systems was assessed. The effects of space environmental conditions on link performance was analyzed. The integration and operation of new fiber optic link components exhibiting increased hardening to radiation environments is emphasized. The opportunity to expose operational fiber optic data links to an actual space environment for 6 months or longer and to retrieve the data links for analysis are discussed. Reliable design criteria for future spacecraft application, relative to the long term low dose space radiation effects as a function of temperature are outlined.

Taylor, E. W.↗

A study of charged particles/radiation damage to VLSI device materials

Future spacecraft systems such as the manned space station will be subjected to low-dose long term radiation particles. Most electronic systems are affected by such particles. There is therefore a great need to understand device physics and failure mechanisms affected by radiation and to design circuits that would be less susceptible to radiation. Using 2 MeV electron radiation and bias temperature aging, it was found that MOS capacitors that were prepositively biased have lower flatband voltage shift and lesser increase in density of surface state charge than those that were not prepositively biased. In addition, it was shown that there is continued recovery of flatband voltage and density of state charge in irradiated capacitors during both room temperature anneal and 137 degree anneal. When nMOS transistors were subjected to 1 MeV proton radiation, charge pumping and current versus voltage measurements indicated that transconductance degradation, threshold voltage shifts and changes in interface states density may be the primary cause of nMOS transistor failure after radiation. Simulation studies using SPICE were performed on CMOS SRAM cells of various transistor sizes. It is shown that transistor sizing affects the noise margins of CMOS SRAM cells, and that as the beta ratio of the transistors of the CMOS SRAM cell decreases, the effective noise margin of the SRAM cell increases. Some suggestions were made in connection with the design of CMOS SRAMS that are hardened against single event upsets.

Okyere, John G.↗

Late cataractogenesis in primates and lagomorphs after exposure to particulate radiations

Results are presented from a study in which regular examinations of cataractogenic development were conducted since 1987 in monkeys that were exposed in 1969, at the age of about 2 years, to low doses of 'mixed-energy' protons (10 and 110 MeV). The results of comparisons with age-matched controls showed that the cataractogenic patterns observed in the exposed monkeys are consistent with those observed in other groups of monkeys that were exposed at similar ages in 1964 and 1965 to protons of different energies (Lett et al., 1986, 1988, 1989, and 1991). Comparisons were also made among recent results from different groups of primates and from New Zealand white rabbits that were exposed when young to Fe-56 and were monitored continuously thereafter.

Cox, A. B.↗

Cellular changes in microgravity and the design of space radiation experiments

Cell metabolism, secretion and cell-cell interactions can be altered during space flight. Early radiobiology experiments have demonstrated synergistic effects of radiation and microgravity as indicated by increased mutagenesis, increased chromosome aberrations, inhibited development, and retarded growth. Microgravity-induced changes in immune cell functions include reduced blastogenesis and cell-mediated, delayed-type hypersensitivity responses, increased cytokine secretions, but inhibited cytotoxic effects an macrophage differentiation. These effects are important because of the high radiosensitivity of immune cells. It is difficult to compare ground studies with space radiation biology experiments because of the complexity of the space radiation environment, types of radiation damage and repair mechanisms. Altered intracellular functions and molecular mechanisms must be considered in the design and interpretation of space radiation experiments. Critical steps in radiocarcinogenesis could be affected. New cell systems and hardware are needed to determine the biological effectiveness of the low dose rate, isotropic, multispectral space radiation and the potential usefulness of radioprotectants during space flight.

Morrison, D. R.↗

Continuous phase and amplitude holographic elements

A method for producing a phase hologram using e-beam lithography provides n-ary levels of phase and amplitude by first producing an amplitude hologram on a transparent substrate by e-beam exposure of a resist over a film of metal by exposing n is less than or equal to m x m spots of an array of spots for each pixel, where the spots are randomly selected in proportion to the amplitude assigned to each pixel, and then after developing and etching the metal film producing a phase hologram by e-beam lithography using a low contrast resist, such as PMMA, and n-ary levels of low doses less than approximately 200 micro-C/sq cm and preferably in the range of 20-200 micro-C/sq cm, and aggressive development using pure acetone for an empirically determined time (about 6 s) controlled to within 1/10 s to produce partial development of each pixel in proportion to the n-ary level of dose assigned to it.

Maker, Paul D.↗

The Impact on Space Radiation Requirements and Effects on ASIMS

The evolution of highly miniaturized electronic and mechanical systems will be accompanied by new problems and issues regarding the radiation response of these systems in the space environment. In this paper we discuss some of the more prominent radiation problems brought about by miniaturization. For example, autonomous micro-spacecraft will require large amounts of high density memory, most likely in the form of stacked, multichip modules of DRAM's, that must tolerate the radiation environment. However, advanced DRAM's (16 to 256 Mbit) are quite susceptible to radiation, particularly single event effects, and even exhibit new radiation phenomena that were not a problem for older, less dense memory chips. Another important trend in micro-spacecraft electronics is toward the use of low-voltage microelectronic systems that consume less power. However, the reduction in operating voltage also caries with it an increased susceptibility to radiation. In the case of application specific integrated microcircuits (ASIM's), advanced devices of this type, such as high density field programmable gate arrays (FPGA's) exhibit new single event effects (SEE), such as single particle reprogramming of anti-fuse links. New advanced bipolar circuits have been shown recently to degrade more rapidly in the low dose rate space environment than in the typical laboratory total dose radiation test used to qualify such devices. Thus total dose testing of these parts is no longer an appropriately conservative measure to be used for hardness assurance. We also note that the functionality of micromechanical Si-based devices may be altered due to the radiation-induced deposition of charge in the oxide passivation layers.

Barnes, C.↗

Fos Expression in Rat Brain During Depletion-Induced Thirst and Salt Appetite

The expression of Fos protein (Fos immunoreactivity, Fos-ir) was mapped in the brain of rats subjected to an angiotensin-dependent model of thirst and salt appetite. The physiological state associated with water and sodium ingestion was produced by the concurrent subcutaneous administration of the diuretic furosemide (10 mg/kg) and a low dose of the angiotensin-converting enzyme (ACE) inhibitor captopril (5 mg/kg; Furo/Cap treatment). The animals were killed 2 h posttreatment, and the brains were processed for Fos-ir to assess neural activation. Furo/Cap treatment significantly increased Fos-ir density above baseline levels both in structures of the lamina terminalis and hypothalamus known to mediate the actions of ANG 2 and in hindbrain regions associated with blood volume and pressure regulation. Furo/Cap treatment also typically increased Fos-ir density in these structures above levels observed after administration of furosemide or captopril separately. Fos-ir was reduced to a greater extent in forebrain than in hindbrain areas by a dose of captopril (100 mg/kg sc) known to block the actions of ACE in the brain. The present work provides further evidence that areas of lamina terminalis subserve angiotensin-dependent thirst and salt appetite.

Thunhorst, R. L.↗