The Predicted Binding Site and Dynamics of Peptide Inhibitors to the Methuselah GPCR from Drosophila melanogaster

Peptide inhibitors of Methuselah (Mth), a G protein-coupled receptor (GPCR), were reported that can extend the life span of Drosophila melanogaster. Mth is a class B GPCR, which is characterized by a large, N-terminal ectodomain that is often involved with ligand recognition. The crystal structure of the Mth ectodomain, which binds to the peptide inhibitors with high affinity, was previously determined. Here we report the predicted structures for RWR motif peptides in complex with the Mth ectodomain. We studied representatives of both Pro-class and Arg-class RWR motif peptides and identified ectodomain residues Asp139, Phe130, Asp127, and Asp78 as critical in ligand binding. To validate these structures, we predicted the effects of various ligand mutations on the structure and binding to Mth. The binding of five mutant peptides to Mth was characterized experimentally by surface plasmon resonance, revealing measured affinities that are consistent with predictions. The electron density map calculated from our MD structure compares well with the experimental map of a previously determined peptide/Mth crystal structure and could be useful in refining the current low-resolution data. The elucidation of the ligand binding site may be useful in analyzing likely binding sites in other class B GPCRs

Non-lethal PCR genotyping of single Drosophila

In Drosophila, genetic techniques relying on stochastic chromosomal rearrangements involve the generation and screening of a large number of fly stocks to isolate a few lines of interest. Here, we describe a PCR-based method allowing non-lethal molecular characterization of single flies. Using this procedure, individual candidate recombinant animals can be genotyped and selected one generation earlier than with extant methodology and, importantly, before stocks are established. This advance should significantly facilitate several of the most fundamental and routine techniques in Drosophila genetics

Mission safety evaluation report for STS-37, postflight edition

STS-37/Atlantis was launched on April 5, 1991 from Kennedy Space Center launch complex 39B at 9:23 a.m. Eastern Standard Time (EST). Launch was delayed 4 minutes 45 seconds because of safety concerns about the low cloud ceiling and the wind direction in the potential blast area. Based on the limited number and type of inflight anomalies encountered, the Space Shuttle operated satisfactorily throughout the STS-37 mission. A contingency EVA was performed by the crew on Flight Day (FD) 3 to free a sticky Gamma Ray Observatory (GRO) high gain antenna, after which the GRO primary payload was successfully deployed by the Orbiter's Remote Manipulator System. The GRO, which weighed just over 35,000 lbs, was the heaviest NASA science satellite ever deployed by the Space Shuttle into low Earth orbit. The scheduled entry/landing on FD 6 was waved off for one day due to high wind conditions at Edwards Air Force Base. Atlantis landed on FD 7, 11 April 1991 on Edwards AFB lakebed runway 33 at 9:55 a.m. Eastern Daylight Time

Examining Trends and Experiences of the Last Four Years of Socially Engineered Ransomware Attacks

This study examines and reports the trends of social engineering-based ransomware attacks over the past four years from three major technology corporations. The focus of the reports was on major threat actors and their attacks against the corporations and their clients. The data were organized chronologically by year, and prevalent or abnormal findings were shared in this report. It was found that social engineering attacks were tremendously effective. Many ransomware attacks stemmed from Phishing. Social engineering approaches allowed attackers to conduct breaches using ransomware applications like GandCrab and Ryuk. Humans are incredibly susceptible and fall for social engineering tactics- primarily Phishing. The information reflected in this report ought to be used to motivate businesses to increase spending and training opportunities for employees to help defend against these social engineering-based attacks

Traffic Controls for Maintenance on High Speed Highways

Observations were first made at lane closures on interstate highways where yellow warning signs were erected routinely in conjunction with contract work. Later data provided direct comparison between new yellow and new orange signs. One sign scheme was used throughout the study. Driver obedience improved when new signs of either color were used; this finding implies that signs should always be maintained in good condition. Orange signs were slightly more effective than yellow signs in reducing traffic conflicts and merges near the traffic cones. The results of the study tend to support the adoption of orange as the standard color for signing construction and maintenance sites. However, the differences between the two colors were rather small. Driver preference polls supported the orange signs more strongly. A degree of driver insensitivity toward signing was shown. In general, variables such as short sight distances, high volumes, poor condition of signs, and driver insensitivity produced unsafe situations at lane closures

Grooving Pavement Centerlines for Lane Demarcation

In late June 1969, the Kentucky Department of Highways contracted to have experimental, longitudinal centerline grooves, 15 feet long at 80-foot intervals, cut into both sets of dual lanes of a 2.7-mile portion of I 71 in Carroll County. When the centerline was painted in an otherwise normal way, the skip lines were alternately on grooved and ungrooved surfaces. The roadway was opened to traffic on July 15, 1969. An interim performance report contained detailed information on the test site, grooving procedures, groove configuration, initial striping efforts, and preliminary evaluations of the relative effectiveness of the grooved stripe. The purposes of this report are: 1) to update the observations cited in the interim report, 2) to evaluate the grooved centerlines during rainy, nighttime conditions, and 3) to record wear and durability histories of grooved and ungrooved lines

Mission safety evaluation report for STS-31, postflight edition

Mission safety factors relative to Space Transportation System (STS) Mission 31 are discussed. In addition to a mission summary, safety risk factors and inflight anomalies are discussed