Studies on the plasmid stability, plasmid copy number and endo(1, 3)(1, 4) b-glucanase production by free and alginate immobilised recombinant saccharomyces cerevisiae cells

A recombinant yeast strain, Saccharomyces cerevisiae DBY746, containing the plasmid pJG317, was grown in a variety of fermentation modes including batch, serial batch and chemostat culture incorporating a wide range of media types Plasmid pJG317 consists of a 2^-denved yeast episomal plasmid containing the gene which encodes for the bacterial enzyme endo (1,3)(1,4) P-glucanase. The concentration of enzyme produced appears to be proportional to the number of plasmid copies per cell. Specific enzyme activities were found to be in the range of 14 x 106 to 48 x lO6 U/cell for free cell culture, with a corresponding plasmid copy number of 8±0 5 to 40±6 3 copies per cell respectively. A procedure for measuring the copy number of pJG317 in S cerevisiae was developed, tested and optimised. The procedure is based on Southern hybridisation and measured the relative intensities of hybridisation of a probe to the single copy yeast chromosomal actin gene and to the multicopy plasmid pJG317. Plasmid pJG317 is quite unstable under non-selective conditions and its copy number and stability are influenced by both growth rate and nutrient supply By immobilising cells in calcium alginate gel beads, the plasmid could be stabilised and high volumetric productivities of up to 38 U/ml-h attained. Although radial gradients in biomass concentration and in percentage of plasmidcontaimng cells in the alginate gel beads were confirmed, no significant difference was found between the plasmid copy number of cells in the centre of the gel beads (36 5±6 8) and cells close to the surface of the gel beads(32 4± 33)

ADR salt pill design and crystal growth process for hydrated magnetic salts

A process is provided for producing a salt pill for use in very low temperature adiabatic demagnetization refrigerators (ADRs). The method can include providing a thermal bus in a housing. The thermal bus can include an array of thermally conductive metal conductors. A hydrated salt can be grown on the array of thermally conductive metal conductors. Thermal conductance can be provided to the hydrated salt

Improved Design and Fabrication of Hydrated-Salt Pills

A high-performance design, and fabrication and growth processes to implement the design, have been devised for encapsulating a hydrated salt in a container that both protects the salt and provides thermal conductance between the salt and the environment surrounding the container. The unitary salt/container structure is known in the art as a salt pill. In the original application of the present design and processes, the salt is, more specifically, a hydrated paramagnetic salt, for use as a refrigerant in a very-low-temperature adiabatic demagnetization refrigerator (ADR). The design and process can also be applied, with modifications, to other hydrated salts. Hydrated paramagnetic salts have long been used in ADRs because they have the desired magnetic properties at low temperatures. They also have some properties, disadvantageous for ADRs, that dictate the kind of enclosures in which they must be housed: Being hydrated, they lose water if exposed to less than 100-percent relative humidity. Because any dehydration compromises their magnetic properties, salts used in ADRs must be sealed in hermetic containers. Because they have relatively poor thermal conductivities in the temperature range of interest (<0.1 K), integral thermal buses are needed as means of efficiently transferring heat to and from the salts during refrigeration cycles. A thermal bus is typically made from a high-thermal-conductivity met al (such as copper or gold), and the salt is configured to make intimate thermal contact with the metal. Commonly in current practice (and in the present design), the thermal bus includes a matrix of wires or rods, and the salt is grown onto this matrix. The density and spacing of the conductors depend on the heat fluxes that must be accommodated during operation

Design of a 3-Stage ADR for the Soft X-Ray Spectrometer Instrument on the Astro-H Mission

The Japanese Astro-H mission will include the Soft X-ray Spectrometer (SXS) instrument, whose 36-pixel detector array of ultra-sensitive x-ray microcalorimeters requires cooling to 50 mK. This will be accomplished using a 3-stage adiabatic demagnetization refrigerator (ADR). The design is dictated by the need to operate with full redundancy with both a superfluid helium dewar at 1.3 K or below, and with a 4.5 K Joule-Thomson (JT) cooler. The ADR is configured as a 2-stage unit that is located in a well in the helium tank, and a third stage that is mounted to the top of the helium tank. The third stage is directly connected through two heat switches to the JT cooler and the helium tank, and manages heat flow between the two. When liquid helium is present, the 2-stage ADR operates in a single-shot manner using the superfluid helium as a heat sink. The third stage may be used independently to reduce the time-average heat load on the liquid to extend its lifetime. When the liquid is depleted, the 2nd and 3rd stages operate as a continuous ADR to maintain the helium tank at as low a temperature as possible - expected to be 1.2 K - and the 1st stage cools from that temperature as a single-stage, single-shot ADR. The ADR s design and operating modes are discussed, along with test results of the prototype 3-stage ADR

Development of a Flight-Worthy 10 to 4 K Continuous Adiabatic Demagnetization Refrigerator

The cryogenics and fluids branch at NASA's Goddard Space Flight Center is currently developing a high-efficiency, vibration-free, flight-worthy Continuous Adiabatic Demagnetization Refrigerator (CADR) that consist of two modular units: one that lifts ~6 microW at 50 mK while rejecting its heat to a 4 K thermal sink, and another unit that provides a constant 4 K cooling stage while rejecting its heat to a thermal sink at 10 K. The two units are linked together via a 4 K common platform. This paper discusses the status report on the thermodynamic performance of the 4 -10 K ADR. This ADR utilizes an Nb3Sn superconducting magnet and Gadolinium Gallium Garnet (GGG) as its refrigerant. Results show that an idealized cycle, one where its hold time at 4 K is equal to the recycle time, can lift 13 mW at 4 K with a hold time of 132 seconds

Development of a Space-Flight ADR Providing Continuous Cooling at 50 Mk with Heat Rejection at 10 K

Future astronomical instruments will require sub-Kelvin detector temperatures to obtain high sensitivity. In many cases large arrays of detectors will be used, and the associated cooling systems will need performance surpassing the limits of present technologies. NASA is developing a compact cooling system that will lift heat continuously at temperatures below 50 mK and reject it at over 10 K. Based on Adiabatic Demagnetization Refrigerators (ADRs), it will have high thermodynamic efficiency and vibration-free operation with no moving parts. It will provide more than 10 times the current flight ADR cooling power at 50 mK and will also continuously cool a 4 K stage for instruments and optics. In addition, it will include an advanced magnetic shield resulting in external field variations below 5 T. We describe the cooling system here and report on the progress in its development

Development of a Space-Flight ADR Providing Continuous Cooling at 50 mK with Heat Rejection at 10 K

Future astronomical instruments will require sub-Kelvin detector temperatures to obtain high sensitivity. In many cases large arrays of detectors will be used, and the associated cooling systems will need performance surpassing the limits of present technologies. NASA is developing a compact cooling system that will lift heat continuously at temperatures below 50 mK and reject it at over 10 K. Based on Adiabatic Demagnetization Refrigerators (ADRs), it will have high thermodynamic efficiency and vibration-free operation with no moving parts. It will provide more than 10 times the current flight ADR cooling power at 50 mK and will also continuously cool a 4 K stage for instruments and optics. In addition, it will include an advanced magnetic shield resulting in external field variations below 5 T. We describe the cooling system here and report on the progress in its development

The Ethiopian Health Extension Program and Variation in Health Systems Performance: What Matters?

Background: Primary health care services are fundamental to improving health and health equity, particularly in the context of low and middle-income settings where resources are scarce. During the past decade, Ethiopia undertook an ambitious investment in primary health care known as the Ethiopian Health Extension Program that recorded impressive gains in several health outcomes. Despite this progress, substantial disparities in health outcomes persist across the country. The objective of this study was to understand how variation in the implementation of the primary health care efforts may explain differences in key health outcomes. Methods and Findings: We conducted a qualitative study of higher-performing and lower-performing woredas using site visits and in-depth interviews undertaken in 7 woredas. We classified woredas as higher-performing or lower-performing based on data on 5 indicators. We conducted a total of 94 open-ended interviews; 12–15 from each woreda. The data were analyzed using the constant comparative method of qualitative data analysis. Substantial contrasts were apparent between higher-performing and lower-performing woredas in use of data for problem solving and performance improvement; collaboration and respectful relationships among health extension workers, community members, and health center staff; and coordination between the woreda health office and higher-level regulatory and financing bodies at the zonal and regional levels. We found similarities in what was reported to motivate or demotivate health extension workers and other staff. Additionally, higher-performing and lower-performing woredas shared concerns about hospitals being isolated from health centers and health posts. Participants from both woredas also highlighted a mismatch between the urban health extension program design and the urban-dwelling communities’ expectations for primary health care. Conclusions: Data-informed problem solving, respectful and supportive relationships with the community, and strong support from zonal and regional health bureaus contributed to woreda performance, suggesting avenues for achieving higher performance in primary health care

Vibration-Heating in ADR Kevlar Suspension Systems

The cryogenics group at NASA's Goddard Space Flight Center has a long-standing development and test program for laboratory and space-flight adiabatic demagnetization refrigerators (ADRs). These devices are used to cool components to temperatures as low as 0.05 K. At such low temperatures the ADR systems can provide a few micro-Watts of cooling power, so it is important to minimize the conduction of heat to these cold stages from the surroundings. The cold ADR elements are held in place by thin tensioned strings made of Kevlar, chosen for its high strength and stiffness and low thermal conductivity. During laboratory testing, we have observed that occasional significant additional heat loads on the coldest ADR stages correlate with unusually high vibration levels in the cryostat due to a noisy mechanical cryocooler. We theorized that this heat results from plastic deformation of the Kevlar fibers and frictional interactions among them, driven by the cryostat vibrations. We describe tests and calculations performed in attempt to confirm this source of the heating, and we discuss possible strategies to reduce this effect in future ADR suspension systems

Continuous Sub-Kelvin Cooling from an Adiabatic Demagnetization Refrigerator

We present the current state of development for a continuous magnetic refrigeration system capable of cooling a detector array or other payload to temperatures below 50 mK. This adiabatic demagnetization refrigerator contains four-stages that are cycling continuously yet present a constant ultra-low temperature heat sink at one physical location within the refrigerator. Two different configurations of essentially the same cooler will be presented where the difference is in the physical layout of the stages and the type of heat sink used for the refrigerator's heat rejection