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At least 775 records · Page 43

The Hyperthermophilic Restriction-Modification Systems of Thermococcus kodakarensis Protect Genome Integrity

Thermococcus kodakarensis (T. kodakarensis), a hyperthermophilic, genetically accessible model archaeon, encodes two putative restriction modification (R-M) defense systems, TkoI and TkoII. TkoI is encoded by TK1460 while TkoII is encoded by TK1158. Bioinformative analysis suggests both R-M enzymes are large, fused methyltransferase (MTase)-endonuclease polypeptides that contain both restriction endonuclease (REase) activity to degrade foreign invading DNA and MTase activity to methylate host genomic DNA at specific recognition sites. In this work, we demonsrate T. kodakarensis strains deleted for either or both R-M enzymes grow more slowly but display significantly increased competency compared to strains with intact R-M systems, suggesting that both TkoI and TkoII assist in maintenance of genomic integrity in vivo and likely protect against viral- or plasmid-based DNA transfers. Pacific Biosciences single molecule real-time (SMRT) sequencing of T. kodakarensis strains containing both, one or neither R-M systems permitted assignment of the recognition sites for TkoI and TkoII and demonstrated that both R-M enzymes are TypeIIL; TkoI and TkoII methylate the N 6 position of adenine on one strand of the recognition sequences GTGAAG and TTCAAG, respectively. Further in vitro biochemical characterization of the REase activities reveal TkoI and TkoII cleave the DNA backbone GTGAAG(N) 20 /(N) 18 and TTCAAG(N) 10 /(N) 8 , respectively, away from the recognition sequences, while in vitro characterization of the MTase activities reveal transfer of tritiated S-adenosyl methionine by TkoI and TkoII to their respective recognition sites. Together these results demonstrate TkoI and TkoII restriction systems are important for protecting T. kodakarensis genome integrity from invading foreign DNA.

59 BASIC BIOLOGICAL SCIENCES↗

Evidence for a cytokine-sensitive network of iron-associated genes that protects pancreatic islets against ferroptosis

Background/Objectives: The micronutrient iron is closely connected to inflammation and is among the complex factors contributing to beta-cell failure in diabetes. High levels of dietary iron increase the risk of developing type 2 diabetes, and excessive iron uptake by beta-cells can cause oxidative stress and inhibit function. Elevated levels of proinflammatory cytokines in obese individuals, such as interleukin (IL)-1beta and IL-6, increase the risk of developing type 2 diabetes, and there is evidence that these low levels of circulating cytokines can lead to islet dysfunction. Methods: In this study, gene microarray and other data were analyzed for expression differences in islets treated for 48 h with 10 pg/mL IL-1beta + 20 pg/mL IL-6 as a model of low-grade inflammation versus untreated. Results: Three iron-associated genes were among the most cytokine-sensitive in the mouse genome: Hamp, Steap4, and Lcn2. These proteins are all involved with increasing/retaining cellular iron. We hypothesized that increased cellular iron would lead to increased susceptibility to ferroptosis. Surprisingly, 24 h pre-exposure to low-grade inflammation, which upregulates this iron-gene network, prevented subsequent erastin-induced ferroptosis. We also found that Steap4 overexpression reduced islet dysfunction caused by high-dose proinflammatory cytokines (10× low-dose), suggesting an overall protective effect. Steap4 overexpression also upregulated Hamp and Lcn2, suggesting Steap4 regulates these cytokine-sensitive iron genes.; in contrast, ferritin and ferroportin gene expression, which are not sensitive to cytokines, were unchanged. Conclusions: These data suggest an inflammation-induced network of genes involved in cellular iron uptake and retention plays a protective role in islets against oxidative stress and ferroptosis.

IL-1β↗

CT584 Is Not a Protective Vaccine Antigen against Respiratory Chlamydial Challenge in Mice

Background: Chlamydia trachomatis is the most prevalent bacterial sexually transmitted pathogen in humans worldwide. Since chlamydial infection is largely asymptomatic with the potential for serious complications, a preventative vaccine is likely the most viable long-term answer to this public health threat. Cell-free protein synthesis (CFPS) utilizes the cellular protein manufacturing machinery decoupled from the requirement for maintaining cellular viability, offering the potential for flexible, rapid, and decentralized production of recombinant protein vaccine antigens. Methods: Here, we use CFPS to produce the full-length putative chlamydial type three secretion system (T3SS) needle-tip protein, CT584, for evaluation as a vaccine antigen in mouse models. High-speed atomic force microscopy (HS-AFM) (RIBM, Tsukuba, Japan) imaging and computer simulations confirm that CFPS-produced CT584 retains a native-like structure prior to immunization. Female mice were primed with CT584 adjuvanted with CpG-1826 intranasally (i.n.) or CpG-1826 + Montanide ISA 720 intramuscularly (i.m.), followed four weeks later by an i.m. boost before respiratory challenge with 10 4 inclusion forming units (IFU) of Chlamydia muridarum. Results: Immunization with CT584 generated robust antibody responses but weak cell-mediated immunity and failed to protect against i.n. challenge as demonstrated by body weight loss, increased lung weights, and the presence of high numbers of IFUs in the lungs. Conclusion: While CT584 was not a protective vaccine candidate, the speed and flexibility with which CFPS can be used to produce other potential chlamydial antigens make it an attractive technique for antigen production.

60 APPLIED LIFE SCIENCES↗

Regulations for Bat Protection in Mexico's Wind Farms

Wind energy development has expanded the fastest globally among all renewable sources during the last 20 years. However, wind farms have documented adverse impacts on bats, including mortality from collisions with turbine blades and disruptions to habitat and behavior. As the world's sixth most attractive economy for renewables, with 70 operating wind farms, Mexico and its bats now face escalating threats from the country's burgeoning wind industry. Despite this rapid growth, few studies have analyzed Mexico's regulatory framework to prevent, evaluate, and mitigate wind farm effects on bats. In this study, we reviewed Mexican laws and treaties that facilitate wind farm permitting, construction, operation, and decommissioning, and searched for guidelines that specifically address bat conservation. We found eight international pacts that promote wind power adoption along with three relevant articles in Mexico's Constitution. The General Law of Ecological Balance and Environmental Protection proved most pertinent for impact management. Supplementary guidelines from the Ministry of Environment and Natural Resources offer general strategies for evaluating wind farm impacts on bats, but adherence remains voluntary. Given expanding wind power investments across Mexico, we highlight the need for more stringent national standards that require preventative and corrective measures to protect bat populations. Tighter legislation and enforcement offer pathways toward environmentally sustainable wind energy development in Mexico.

17 WIND ENERGY↗

Protection Against Radiation Hazards in Space, Book I Proceedings of the Symposium at Gatlinburg, Tenn., Nov. 5-7, 1962

BOOK 1: Radiation Environment in Space Effects of Space Radiation on Radiosensitive Objects Biological Effects of Space Radiation BOOK 2 : Shielding Against Space Radiations The realization in recent years that outer space is traversed by high-energy radiations has caused man to reevaluate the feasibility of manned or even instrumented exploration outside our atmosphere. Fortunately, it is possible to determine the nature and intensities of these radiations and to produce similar radiations on earth by means of accelerators. Thus we can learn how to attenuate them and to design capsules which afford protection against them. Of course this protection carries a weight penalty so that there is a premium on optimizing the shield design. Many groups in the United States are engaged in research to this end, and it was the purpose of this symposium to bring these groups together so that they could exchange information. To make the meeting more comprehensive, sessions on the nature of the radiations and their effects on people and things were included. However, the major part of the meeting was devoted to discussions on shielding research, comprising theoretical calculations and experiments carried out mainly with high-energy accelerators. The symposium committee feels that the aims of the symposium were met and that progress in space research program was greatly accelerated thereby.

RADIATION SHIELDING↗

Proceedings of the Symposium on the Protection Against Radiation Hazards in Space

Shielding Against Space Radiations The realization in recent years that outer space is traversed by high-energy radiations has caused man to reevaluate the feasibility of manned or even instrumented exploration outside our atmosphere. Fortunately, it is possible to determine the nature and intensities of these radiations and to produce similar radiations on earth by means of accelerators. Thus we can learn how to attenuate them and to design capsules which afford protection against them. Of course this protection carries a weight penalty so that there is a premium on optimizing the shield design. Many groups in the United States are engaged in research to this end, and it was the purpose of this symposium to bring these groups together so that they could exchange information. To make the meeting more comprehensive, sessions on the nature of the radiations and their effects on people and things were included. However, the major part of the meeting was devoted to discussions on shielding research, comprising theoretical calculations and experiments carried out mainly with high-energy accelerators. The symposium committee feels that the aims of the symposium were met and that progress in space research program was greatly accelerated thereby.

SECONDARY EMISSION↗

Epoxy blanket protects milled part during explosive forming

Epoxy blanket protects chemically milled or machined sections of large, complex structural parts during explosive forming. The blanket uniformly covers all exposed surfaces and fills any voids to support and protect the entire part.

Source record↗

Run-in with chemical additive protects gear surface

Run-in treatment provides a protective coating on turbopump gear surfaces so that they are capable of operation under marginal conditions in mineral oil and diester lubricants. This treatment protects highly loaded gears during relatively short-term operation.

Hartman, M. A.↗

Protected, high-temperature connecting cable

Ceramic insulated, swaged stainless steel, sheathed, protective atmosphere cable admits electrical leads into an 1800 deg F air-environment test chamber. The cable has some bending capability and provides for nine niobium alloy conductors. An argon purge during the TIG weld closure protects internal wires from oxidation and embrittlement.

Engdahl, R. E.↗

Eutectic fuse provides current and thermal protection under high vibration

Eutectic fuses provide current and thermal protection to an electronic system and maintain this protection under high vibration environments. The fuses are embedded within heat shrinkable sleeving which provides positive closing action under the conditions of high current or temperature.

Ierokomos, N.↗

Air sampler collects and protects minute particles

Air ejector impactor sampler collects and protects samples of particles greater than 0.1 micron in diameter. In operation, it causes impaction of particle-laden air onto several collection surfaces within a collection cylinder. When not operating, the collector cylinder is maintained in a retracted state within a protective envelope.

Wood, R. C.↗

Fuse protects circuit from voltage and current overloads

Low-melting resistor connected in series with the load protects the circuit against current overloads. It protects test subjects and patients being monitored by electronic instrumentation from inadvertant overloads of current, and sensitive electronic equipment against high-voltage damage.

Casey, L. O.↗

Evaluation of heat- and blast-protection materials

A program was initiated at the Kennedy Space Center in December 1967 and conducted through December 1969 to evaluate the performance of heat- and blast-protection materials for ground support equipment used during the Apollo/Saturn launches. Materials believed to be generally suitable for heat and blast protection were subjected to launch-exposure tests. Tests were made during the Apollo/Saturn 502, 503, and 505 launches. Tests were also made in a local laboratory, as an alternative to the restrictive requirements of launch-exposure tests, to determine the effects of torch-flame exposure on ablative materials. Five materials were found to be satisfactory in all major test categories. It was determined that torch-flame tests can probably be utilized as an acceptable substitute for the booster-engine-exhaust exposure tests for basic screening of candidate materials.

Morrison, J. D.↗

Apollo experience report: Protection of life and health

The development, implementation, and effectiveness of the Apollo Lunar Quarantine Program and the Flight Crew Health Stabilization Program are discussed as part of the broad program required for the protection of the life and health of U.S. astronauts. Because the goal of the Apollo Program has been the safe transport of men to the moon and back to earth, protection of the astronauts and of the biosphere from potentially harmful lunar contaminants has been required. Also, to ensure mission success, the continuing good health of the astronauts before and during a mission has been necessary. Potential applications of specific aspects of the health and quarantine programs to possible manned missions to other planets are discussed.

Wooley, B. C.↗

Advanced Extravehicular Protective System (AEPS) study

A summary is presented of Advanced Extravehicular Protective Systems (AEPS) for the future missions beyond Skylab in earth orbit, on the lunar surface, and on the Martian surface. The study concentrated on the origination of regenerable life support concepts for use in portable extravehicular protective systems, and included evaluation and comparison with expendable systems, and selection of life support subsystems. The study was conducted in two phases. In the first phase, subsystem concepts for performing life support functions in AEPS which are regenerable or partially regenerable were originated, and in addition, expendable subsystems were considered. Parametric data for each subsystem concept were evolved including subsystem weight and volume, power requirement, thermal control requirement; base regeneration equipment weight and volume, requirement. The second phase involved an evaluation of the impact of safety considerations involving redundant and/or backup systems on the selection of the regenerable life support subsystems. In addition, the impact of the space shuttle program on regenerable life support subsystem development was investigated.

Williams, J. L.↗

Protective coatings for columbium applied in the field.

The various aspects of field repair of columbium alloy panels with protective coatings designed as part of the Space Shuttle thermal protection system are examined. The field repair of the coatings is accomplished by employing ceramic cement repairs, and reapplying the fused slurry silicide coating. Techniques are described which improve the practicality of these repairs by employing torch heating. The repair coating quality is demonstrated by testing which simulates flight temperature, pressure, stress and acoustic vibration conditions as a function of time. Conclusions on the present status of field repair coatings are presented and recommendations are given for appropriate future activities relative to the use on an operational Space Shuttle system.

Carter, J.↗