Repetitive Segmental Structure of the Transducin β Subunit: Homology with the CDC4 Gene and Identification of Related mRNAs

Retinal transducin, a guanine nucleotide regulatory protein (referred to as a G protein) that activates a cGMP phosphodiesterase in photoreceptor cells, is comprised of three subunits. We have identified and analyzed cDNA clones of the bovine transducin β subunit that may be highly conserved or identical to that in other G proteins. From the cDNA nucleotide sequence of the entire coding region, the primary structure of a 340-amino acid protein was deduced. The encoded β subunit has a Mr of 37,375 and is comprised of repetitive homologous segments arranged in tandem. Furthermore, significant homology in primary structure and segmental sequence exists between the β subunit and the yeast CDC4 gene product. The Mr 37,375 β subunit polypeptide is encoded by a 2.9-kilobase (kb) mRNA. However, there exists in retina other β-related mRNAs that are divergent from the 2.9-kb mRNA on the basis of oligonucleotide and primer-extended probe hybridizations. All mammalian tissues and clonal cell lines that have been examined contain at least two β-related mRNAs, usually 1.8 and 2.9 kb in length. These results suggest that the mRNAs are the processed products of a small number of closely related genes or of a single highly complex β gene

Energy stress-mediated cytotoxicity in tuberous sclerosis complex 2-deficient cells with nelfinavir and mefloquine treatment

To find new anti-cancer drug therapies, we wanted to exploit homeostatic vulnerabilities within Tuberous Sclerosis Complex 2 (TSC2)-deficient cells with mechanistic target of rapamycin complex 1 (mTORC1) hyperactivity. We show that nelfinavir and mefloquine synergize to selectively evoke a cytotoxic response in TSC2-deficient cell lines with mTORC1 hyperactivity. We optimize the concentrations of nelfinavir and mefloquine to a clinically viable range that kill cells that lack TSC2, while wild-type cells tolerate treatment. This new clinically viable drug combination causes a significant level of cell death in TSC2-deficient tumor spheroids. Furthermore, no cell recovery was apparent after drug withdrawal, revealing potent cytotoxicity. Transcriptional profiling by RNA sequencing of drug treated TSC2-deficient cells compared to wild-type cells suggested the cytotoxic mechanism of action, involving initial ER stress and an imbalance in energy homeostatic pathways. Further characterization revealed that supplementation with methyl pyruvate alleviated energy stress and reduced the cytotoxic effect, implicating energy deprivation as the trigger of cell death. This work underpins a critical vulnerability with cancer cells with aberrant signaling through the TSC2-mTORC1 pathway that lack flexibility in homeostatic pathways, which could be exploited with combined nelfinavir and mefloquine treatment

Two Messages from the President of the United States Communicating Additional Correspondence in Relation to the Adjustment of the Northeastern Boundary, and the Occupation of the Disputed Territory

Correspondence authored by President Van Buren, American Secretary of State, John Forsyth, Maine Governor, John Fairfield, British Envoy Henry S. Fox, and others regarding the occupation and movement of British soldiers in the disputed territory along the northeastern boundary of the State of Maine between Maine and modern-day New Brunswick, Canada. The border issue was resolved on August 9, 1842 with the Webster-Ashburton Treaty.https://digitalcommons.library.umaine.edu/mainebicentennial/1051/thumbnail.jp

The N2K Consortium. II. A Transiting Hot Saturn Around HD 149026 With a Large Dense Core

Doppler measurements from Subaru and Keck have revealed radial velocity variations in the V=8.15, G0IV star HD 149026 consistent with a Saturn-Mass planet in a 2.8766 day orbit. Photometric observations at Fairborn Observatory have detected three complete transit events with depths of 0.003 mag at the predicted times of conjunction. HD 149026 is now the second brightest star with a transiting extrasolar planet. The mass of the star, based on interpolation of stellar evolutionary models, is 1.3 +/- 0.1 solar masses; together with the Doppler amplitude, K=43.3 m s^-1, we derive a planet mass Msin(i)=0.36 Mjup, and orbital radius of 0.042 AU. HD 149026 is chromospherically inactive and metal-rich with spectroscopically derived [Fe/H]=+0.36, Teff=6147 K, log g=4.26 and vsin(i)=6.0 km s^-1. Based on Teff and the stellar luminosity of 2.72 Lsun, we derive a stellar radius of 1.45 Rsun. Modeling of the three photometric transits provides an orbital inclination of 85.3 +/- 1.0 degrees and (including the uncertainty in the stellar radius) a planet radius of 0.725 +/- 0.05 Rjup. Models for this planet mass and radius suggest the presence of a ~67 Mearth core composed of elements heavier than hydrogen and helium. This substantial planet core would be difficult to construct by gravitational instability.Comment: 25 pages, 5 figures, accepted by the Astrophysical Journa

Structure of a putative NTP pyrophosphohydrolase: YP_001813558.1 from Exiguobacterium sibiricum 255-15.

The crystal structure of a putative NTPase, YP_001813558.1 from Exiguobacterium sibiricum 255-15 (PF09934, DUF2166) was determined to 1.78 Å resolution. YP_001813558.1 and its homologs (dimeric dUTPases, MazG proteins and HisE-encoded phosphoribosyl ATP pyrophosphohydrolases) form a superfamily of all-α-helical NTP pyrophosphatases. In dimeric dUTPase-like proteins, a central four-helix bundle forms the active site. However, in YP_001813558.1, an unexpected intertwined swapping of two of the helices that compose the conserved helix bundle results in a `linked dimer' that has not previously been observed for this family. Interestingly, despite this novel mode of dimerization, the metal-binding site for divalent cations, such as magnesium, that are essential for NTPase activity is still conserved. Furthermore, the active-site residues that are involved in sugar binding of the NTPs are also conserved when compared with other α-helical NTPases, but those that recognize the nucleotide bases are not conserved, suggesting a different substrate specificity

The N2K Consortium. I. A Hot Saturn Planet Orbiting HD 88133

The N2K ("next 2000") consortium is carrying out a distributed observing campaign with the Keck, Magellan, and Subaru telescopes, as well as the automatic photometric telescopes of Fairborn Observatory, in order to search for short-period gas giant planets around metal-rich stars. We have established a reservoir of more than 14,000 main-sequence and subgiant stars closer than 110 pc, brighter than V = 10.5, and with 0.4 0.1 dex for this survey. We outline the strategy and report the detection of a planet orbiting the metal-rich G5 IV star HD 88133 with a period of 3.41 days, semivelocity amplitude K = 35.7 m s^(-1), and M sin i = 0.29M_J. Photometric observations reveal that HD 88133 is constant on the 3.415 day radial velocity period to a limit of 0.0005 mag. Despite a transit probability of 19.5%, our photometry rules out the shallow transits predicted by the large stellar radius

Small UAS Detect and Avoid Requirements Necessary for Limited Beyond Visual Line of Sight (BVLOS) Operations

Potential small Unmanned Aircraft Systems (sUAS) beyond visual line of sight (BVLOS) operational scenarios/use cases and Detect And Avoid (DAA) approaches were collected through a number of industry wide data calls. Every 333 Exemption holder was solicited for this same information. Summary information from more than 5,000 exemption holders is documented, and the information received had varied level of detail but has given relevant experiential information to generalize use cases. A plan was developed and testing completed to assess Radio Line Of Sight (RLOS), a potential key limiting factors for safe BVLOS ops. Details of the equipment used, flight test area, test payload, and fixtures for testing at different altitudes is presented and the resulting comparison of a simplified mathematical model, an online modeling tool, and flight data are provided. An Operational Framework that defines the environment, conditions, constraints, and limitations under which the recommended requirements will enable sUAS operations BVLOS is presented. The framework includes strategies that can build upon Federal Aviation Administration (FAA) and industry actions that should result in an increase in BVLOS flights in the near term. Evaluating approaches to sUAS DAA was accomplished through five subtasks: literature review of pilot and ground observer see and avoid performance, survey of DAA criteria and recommended baseline performance, survey of existing/developing DAA technologies and performance, assessment of risks of selected DAA approaches, and flight testing. Pilot and ground observer see and avoid performance were evaluated through a literature review. Development of DAA criteria—the emphasis here being well clear— was accomplished through working with the Science And Research Panel (SARP) and through simulations of manned and unmanned aircraft interactions. Information regarding sUAS DAA approaches was collected through a literature review, requests for information, and direct interactions. These were analyzed through delineation of system type and definition of metrics and metric values. Risks associated with sUAS DAA systems were assessed by focusing on the Safety Risk Management (SRM) pillar of the SMS (Safety Management System) process. This effort (1) identified hazards related to the operation of sUAS in BVLOS, (2) offered a preliminary risk assessment considering existing controls, and (3) recommended additional controls and mitigations to further reduce risk to the lowest practical level. Finally, flight tests were conducted to collect preliminary data regarding well clear and DAA system hazards