Localization of sequences regulating ancestral and acquired sites of esterase6 activity in Drosophila melanogaster

We have broadly defined the DNA regions regulating esterase6 activity in several life stages and tissue types of D. melanogaster using P- element-mediated transformation of constructs that contain the esterase6 coding region and deletions or substitutions in 5' or 3' flanking DNA. Hemolymph is a conserved ancestral site of EST6 activity in Drosophila and the primary sequences regulating its activity lie between -171 and -25 bp relative to the translation initiation site: deletion of these sequences decrease activity approximately 20-fold. Hemolymph activity is also modulated by four other DNA regions, three of which lie 5' and one of which lies 3' of the coding region. Of these, two have positive and two have negative effects, each of approximately twofold. Esterase6 activity is present also in two male reproductive tract tissues; the ejaculatory bulb, which is another ancestral activity site, and the ejaculatory duct, which is a recently acquired site within the melanogaster species subgroup. Activities in these tissues are at least in part independently regulated: activity in the ejaculatory bulb is conferred by sequences between -273 and -172 bp (threefold decrease when deleted), while activity in the ejaculatory duct is conferred by more distal sequences between -844 and -614 bp (fourfold decrease when deleted). The reproductive tract activity is further modulated by two additional DNA regions, one in 5' DNA (-613 to -284 bp; threefold decrease when deleted) and the other in 3' DNA (+1860 to +2731 bp; threefold decrease when deleted) that probably overlaps the adjacent esteraseP gene. Collating these data with previous studies suggests that expression of EST6 in the ancestral sites is mainly regulated by conserved proximal sequences while more variable distal sequences regulate expression in the acquired ejaculatory duct site.      

Different dose regimens of a SARS-CoV-2 recombinant spike protein vaccine (NVX-CoV2373) in younger and older adults: A phase 2 randomized placebo-controlled trial

BackgroundNVX-CoV2373 is a recombinant severe acute respiratory coronavirus 2 (rSARS-CoV-2) nanoparticle vaccine composed of trimeric full-length SARS-CoV-2 spike glycoproteins and Matrix-M1 adjuvant.Methods and findingsThe phase 2 component of our randomized, placebo-controlled, phase 1 to 2 trial was designed to identify which dosing regimen of NVX-CoV2373 should move forward into late-phase studies and was based on immunogenicity and safety data through Day 35 (14 days after the second dose). The trial was conducted at 9 sites in Australia and 8 sites in the United States. Participants in 2 age groups (aged 18 to 59 and 60 to 84 years) were randomly assigned to receive either 1 or 2 intramuscular doses of 5-μg or 25-μg NVX-CoV2373 or placebo, 21 days apart. Primary endpoints were immunoglobulin G (IgG) anti-spike protein response, 7-day solicited reactogenicity, and unsolicited adverse events. A key secondary endpoint was wild-type virus neutralizing antibody response. After enrollment, 1,288 participants were randomly assigned to 1 of 4 vaccine groups or placebo, with 1,283 participants administered at least 1 study treatment. Of these, 45% were older participants 60 to 84 years. Reactogenicity was predominantly mild to moderate in severity and of short duration (median <3 days) after first and second vaccination with NVX-CoV2373, with higher frequencies and intensity after second vaccination and with the higher dose. Reactogenicity occurred less frequently and was of lower intensity in older participants. Both 2-dose regimens of 5-μg and 25-μg NVX-CoV2373 induced robust immune responses in younger and older participants. For the 2-dose regimen of 5 μg, geometric mean titers (GMTs) for IgG anti-spike protein were 65,019 (95% confidence interval (CI) 55,485 to 76,192) and 28,137 (95% CI 21,617 to 36,623) EU/mL and for wild-type virus neutralizing antibody (with an inhibitory concentration of 50%—MN_50%) were 2,201 (95% CI 1,343 to 3,608) and 981 (95% CI 560 to 1,717) titers for younger and older participants, respectively, with seroconversion rates of 100% in both age groups. Neutralizing antibody responses exceeded those seen in a panel of convalescent sera for both age groups. Study limitations include the relatively short duration of safety follow-up to date and current lack of immune persistence data beyond the primary vaccination regimen time point assessments, but these data will accumulate over time.ConclusionsThe study confirmed the phase 1 findings that the 2-dose regimen of 5-μg NVX-CoV2373 is highly immunogenic and well tolerated in younger adults. In addition, in older adults, the 2-dose regimen of 5 μg was also well tolerated and showed sufficient immunogenicity to support its use in late-phase efficacy studies.Trial registrationClinicalTrials.gov{"type":"clinical-trial","attrs":{"text":"NCT04368988","term_id":"NCT04368988"}}NCT04368988.

In a phase 2 randomized placebo-controlled trial, Neil Formica and coauthors investigate the immunogenicity and safety of different dose regimens of a SARS-CoV-2 recombinant spike protein vaccine in younger and older adults in USA and Australia.     

Numerical taxonomic analysis of cross-inoculation patterns of legumes and Rhizobium

Summary A survey was made of published results of tests of the capacity of Rhizobium derived from one legume genus to nodulate plants from other genera. The data were derived from more than 14,000 separate cross-inoculation trials involving species from 165 genera of legumes. Numerical taxonomic techniques were applied to 113 of the genera for which results of substantial cross-infection tests were available. The data were examined using mean character difference coefficients re-expressed as total and positive-only similarity coefficients. The resulting similarity matrices were clustered by the unweighted pair-group method using arithmetic averages. Eighteen affinity groups were defined at the 70% similarity level. With few exceptions, the physiological and cultural behavior of the rhizobia was consistent within the defined groups. Two broad categories were suggested in the numerical taxonomic analysis, and their validity is discussed in regard to the geographic, physiological and cultural characteristics of the legumes and their Rhizobium microsymbionts. The taxonomic and agronomic value of this approach and the new groupings are discussed.     

Confirmation of a wintering ground of Ross's Gull Rhodostethia rosea in the northern Labrador Sea

Ross's Gull Rhodostethia rosea is one of the world's least known seabirds; < 1% of the estimated global population can be accounted for at known breeding sites, and its wintering range has never been determined. Anecdotal reports over the last two centuries have prompted extensive speculation as to possible wintering areas used by this species in the north Pacific/Bering Sea region, but none has ever been confirmed. Using satellite and geolocator telemetry, we show that some Ross's Gulls from a colony in the Canadian Arctic winter in a restricted area of the northern Labrador Sea. Our discovery of a wintering area in the northwest Atlantic indicates that Ross's Gulls breeding in the Nearctic may be part of a disjunct population, or that birds breeding in the Palaearctic may winter off the east coast of North America.     

Application of FAME (fatty acid methyl ester) analysis in the numerical taxonomic determination of bacterial guild structure

Comparative numerical taxonomic analyses, using fatty acid methyl ester (FAME) profiles and phenetic characteristics, were conducted to examine bacterial guild structure in freshwater sediments. Both approaches were used to examine a subset of 60 OTUs obtained from a previously well characterized microbial community in sediments of a shallow fast flowing stream. For both classifications, greater than 80% of the OTUs were recovered in 11 and 12 major groups for FAME and phenetics approaches. However, there was not complete correspondence for the groupings of the 2 classifications, with most FAME groups being distributed among the phenetic groups and 2 phenetic groups not providing usable characteristics for FAME analysis. The results did demonstrate significant taxonomic variation in bacteria capable of occupying the same or a similar fundamental niche.     

A graphical method for detecting recombination in phylogenetic data sets

Current phylogenetic tree reconstruction methods assume that there is a single underlying tree topology for all sites along the sequence. The presence of mosaic sequences due to recombination violates this assumption and will cause phylogenetic methods to give misleading results due to the imposition of a single tree topology on all sites. The detection of mosaic sequences caused by recombination is therefore an important first step in phylogenetic analysis. A graphical method for the detection of recombination, based on the least squares method of phylogenetic estimation, is presented here. This method locates putative recombination breakpoints by moving a window along the sequence. The performance of the method is assessed by simulation and by its application to a real data set.      

Evaluating Sierra Nevada Yellow-Legged Frog Distribution Using Environmental DNA

Genetic tools that identify species from trace DNA samples could supplement traditional survey methods to clarify distributional limits of rare species. For species with legal habitat protection, elevational limits of distributions are used to determine where management actions may affect endangered species. The endangered Sierra Nevada yellow-legged frog (Rana sierrae) generally is found down to 1,370 m, but in the Plumas National Forest, California, USA, there are a number of historical records below this elevation, resulting in protections extending to 1,067 m. This species is phenotypically similar to the foothill yellow-legged frog (R. boylii), with which it occasionally hybridizes. We used a combination of genetic methods to investigate the fine-scale distribution of the Sierra Nevada yellow-legged frog in the Plumas National Forest. We collected and analyzed environmental DNA (eDNA) samples from all accessible lower elevation sites with records of Sierra Nevada yellow-legged frog (n = 17) and swabbed 220 individuals for genetic identification from 2016–2018 to clarify the distribution of this endangered species. We created a climatic suitability model using the validated Sierra Nevada yellow-legged frog records and current (1970–2000) climate models to assess additional highly suitable localities for Sierra Nevada yellow-legged frog presence using eDNA capture. We did not confirm detection of Sierra Nevada yellow-legged frog eDNA at any historical sites and identified all swabbed individuals from below 1,370 m (n = 144) as foothill yellow-legged frogs. We located a new Sierra Nevada yellow-legged frog site (at 1,919 m) during surveys guided by the climatic suitability model. It does not appear after extensive eDNA and genetic sampling that the Sierra Nevada yellow-legged frog occurs below 1,370 m in this portion of their range at present. Our results show that eDNA sampling can be used as an effective management tool to evaluate historical locations and previously unknown suitable localities for current presence of a species of interest. © 2021 The Authors. The Journal of Wildlife Management published by Wiley Periodicals LLC on behalf of The Wildlife Society.