Category Archives: Evolution

It has been 186 years since Charles Darwin collected the samples of the Galapagos Islands species that led to his explanation of how the diversity of life on Earth has evolved and forever changed the way we understand the world.

During his five-week stay on the islands, Darwin collected dozens of samples, including one small, light brownish-grey snake on Floreana Island. That sample, now at the Natural History Museum in London, was the basis for describing a new species, the Galapagos (Floreana) racer.

The species disappeared from Floreana but can still be found on two satellite islands. Now, UC Merceds evolutionary biology and conservation genetics Professor Danielle Edwards and her research group are the first scientists to propose genetically sequencing Darwins original Galapagos racer sample. Edwards recently got a 202021 Research Publication Grant in Engineering, Medicine, and Science Grant through the American Association of University Women (AAUW) to study Galapagos snakes.

We want to effect ecosystem restoration on Floreana Island by finding an evolutionary replacement in the ecosystem for that species. To do this, we will sequence Darwin's specimen to understand the identity of the specimen relative to the populations present on other islands. We can then use the closest genetic relative to repopulate Floreana Island with this key predator, Edwards said. "We will also use this dataset to understand the extent of species diversity across the archipelago with the most extensive sampling to date."

Anecdotally, racer snakes have fallen prey to cats and other invasive species across the islands. Edwards and her team will use genetic data gathered from samples collected by Galapagos National Park and many international collaborators, along with specimens from museums, to assess population size changes in racer populations and see if populations are in decline, or stable, informing future conservation management efforts by the Galapagos National Park.

Its a huge effort, involving an international team of collaborators, and we do not yet know if we will be able to get DNA from the specimen Darwin collected, Edwards said. But we wont know unless we try, and results from other similar studies on museum specimens have shown promising results.

The Galapagos is a prime place to undertake conservation and rewilding research, because the park has an extensive record of incorporating rigorous science into practices with many partner organizations, she said.

Edwards will lead a team including colleagues from the United States, United Kingdom, Ecuador and New Zealand, and the Galapagos National Park and Island Conservation to develop the project. They have been collecting samples for the past five years to try and identify novel species on the islands and expand sampling to include all known islands and islets where snakes occur.

The findings of this research will provide a detailed picture of how snakes evolved across the islands. The snakes are the last reptile group to have their historical movements across the islands studied. They are believed to be the most mobile of the terrestrial vertebrates and are therefore more likely to provide insights into connections among islands, Edwards said. This project will also provide a source population for Floreana rewilding and a detailed assessment of the genetic health of snake populations across the islands.

Edwards, who joined the Department of Life and Environmental Sciences in the School of Natural Sciences in 2015, focuses most of her research on how the environment impacts the evolution of ecological niche, phenotype and behavior in reptiles, mostly in the context of how these changes lead to the development of new species. She applies this research to inform conservation management strategies for endangered and vulnerable reptiles and amphibians.

But this isnt her first genetics project in the Galapagos. In 2015, a team she was part of revealed that it had identified a novel species of Galapagos giant tortoise. Edwards conducted much of the population genetics analyses, using repeat fingerprinting markers like those used in forensic research, that let the team distinguish between two closely related species. She has been involved in projects in the Galapagos and Australia, applying genomic techniques to understanding biodiversity and implementing conservation management over the past 20 years.

Working in the Galapagos is a dream come true for an evolutionary biologist, herpetologist and conservation biologist, she said. I feel privileged and excited to work on a project based in the crucible of evolutionary thought with a specimen Darwin himself collected.

Here is the original post:

Edwards Eager to Expand Father of Evolution's Work | Newsroom - UC Merced University News

New Zealands South Island was once covered in thick forest, the trees breaking like a dark green tide around the grassy mountaintops. After the arrival of Maori settlers about 750 years ago, some hillsides were cleared of their trees by humans using fire, and the foliage has not returned. For the organisms living in these forests, their habitat changed nearly overnight from sheltered woodland to exposed, windy grasslands.

Since the forests burned, little winged insects called stoneflies have changed as well, researchers have found. In a kind of evolutionary pivot over the course of just a handful of centuries, the stoneflies living above the tree line have lost the ability to fly, suggesting that human-made changes to an ecosystem, such as deforestation, can radically reshape the bodies of its inhabitants. The discovery was published in the journal Biology Letters.

Charles Darwin noticed that insects on islands have a curious tendency to be flightless, perhaps because flying is dangerous when you are tiny and winds are strong. In New Zealand, scientists had found flightless stoneflies on many different mountains, said Jon Waters, a professor at the University of Otago in New Zealand who is an author of the new paper. It was not initially clear why whether there was something about the altitude that favored a flightless form, or if there was something else going on.

To answer the question, he and his colleagues collected stoneflies at five sites, walking up through the forests onto the bald crests of the mountains. They caught insects as they went up the slopes, recording their locations. Looking at all the data, they were surprised to find a very clear trend.

We found that there was this amazing transition from winged populations to flightless populations as you go up, said Waters. Wherever we looked, that correlation was linked to where the trees stopped, not a particular altitude.

Because the switch happened at the tree line, rather than at a specific elevation, it suggests that the exposed situation above the trees has favored flightlessness in stoneflies. Perhaps, as in the case of Darwins island insects, wind on the heights makes flying a liability.

It is possible that in some places, even before the forests burned, there were already flightless stoneflies that simply expanded their territory after the fires. A genetic analysis of the stoneflies showed that three of the five populations the researchers looked at were quite different from their lowland winged brethren, implying that they may have been evolving on their own for a while.

The other two, however, had smaller differences, suggesting the change might be recent recent enough to be since humans arrived on the island.

The apparent swiftness of the change recalls the case of the peppered moth, whose coloration shifted from light to dark as air pollution from the Industrial Revolution in England darkened the trees it lived on; light moths, of course, were more visible to predators in their changed environment. It does not take millennia for animal populations to be altered by natural selection, these cases show.

You go into the trees, and youve suddenly gone into a different population. Its almost like magic that evolution seems to be working so clearly and so effectively over a short distance in some of these cases and a short time frame, Waters said.

Now the researchers are looking deeper into the genetics of the stoneflies to understand what it is that changes as the insects lose the ability to fly. The details may reveal whether stoneflies apparent flexibility arises from new mutations, or whether their flightlessness draws on variations that already existed in their ancestral populations and was just waiting for the right moment to take over.

This article originally appeared in The New York Times.

View original post here:

When insects lost their homes, evolution clipped their wings - The Indian Express

HOUSTON, TX / ACCESSWIRE / August 16, 2021 / Evolution Petroleum (NYSE American:EPM) ("Evolution" or the "Company") today announced its reserves as determined by its independent reserve engineer, DeGolyer & MacNaughton, for the fiscal year ended June 30, 2021.

Highlights:

Diversified its portfolio through the acquisition of long-life producing assets in the Barnett Shale adding 13.1 million barrels of oil equivalent ("MMBOE") to its reserves

Substantially increased fiscal year-end proved reserves to 23.4 MMBOE from 10.2 MMBOE at fiscal year-end 2020

Positive reserve revisions net of production due primarily to the return to service of the Delhi CO2 supply line and well reactivations at Hamilton Dome

Reserves based on trailing twelve-month commodity prices that are substantially below current prices

Jason Brown, President and Chief Executive Officer, commented, "We were pleased to more than double our fiscal 2021 year-end proved reserves with the acquisition of a non-operated ownership interest in Barnett Shale natural gas assets in May 2021. The associated proved developed producing (PDP) reserves are anchored by a low annual decline of less than 10% and include potential upside through workover opportunities. The acquisition of the Barnett Shale assets helped to diversify our production mix to include natural gas with favorable access to the Gulf Coast markets that offer premium pricing. Additionally, we had positive reserve revisions this year at Delhi and Hamilton Dome, further demonstrating the strength and value of our existing long-life producing assets. We were encouraged by the return to service of the CO2 supply line to Delhi in late 2020 that will allow resumption of normal injection levels and provide meaningful reservoir pressure support for the continued recovery and development of the field's long-life reserves. At Hamilton Dome, the operator was able to return substantially all of the wells shut-in due to the dramatic price drop caused by the pandemic, adding to production and extending the life of the field. Hamilton Dome reserves were also revised upward due to improved year-over-year pricing. We look forward to continuing to benefit from the improved price environment as we evaluate and execute additional opportunities to further grow our asset base and reserves."

Story continues

As of June 30, 2021

Oil

NGL

Natural Gas

Equivalent

MBO

MBL

MMcf

MBOE*

Proved Developed Producing

6,815

6,663

48,571

21,573

92

%

Proved Undeveloped

1,605

209

-

1,814

8

%

Total Proved

8,420

6,872

48,571

23,387

100

%

Probable Developed Producing

2,052

743

-

2,795

85

%

Probable Undeveloped

489

-

-

489

15

%

Total Probable**

2,541

743

-

3,284

100

%

Possible Developed Producing

2,251

314

-

2,565

91

%

Possible Undeveloped

255

-

-

255

9

%

Total Possible**

2,506

314

-

2,820

100

%

* Equivalent reserves based on a ratio of 6 Mcf of natural gas to 1 Bbl of oil.

** Read the section captioned "Cautionary Statement" later in this release addressing reserves.

Proved Reserves

The Company's year ended June 30, 2021 SEC proved reserves were 23.4 MMBOE, up from 10.2 MMBOE at year ended June 30, 2020. Although both Delhi and Hamilton Dome had positive revisions net of production, the majority of the increase was due to the acquisition of non-operated interests in the Barnett Shale in May 2021 that added 48.6 Bcf of natural gas, 4.9 MMBL of natural gas liquids, and 0.1 MMBO of oil proved reserves (13.1 MMBOE).

The SEC pricing (twelve-month first day of the month average realized prices) used in the report for the year ended June 30, 2021 was $49.72 per barrel of oil and $2.46 per MCF of natural gas. For fiscal year ended June 30, 2020, SEC average realized prices were $47.37 per barrel of oil and $2.12 per MCF of natural gas.

Approximately 65% of year ended June 30, 2021 SEC proved reserves were liquids (36% crude oil and 29% NGLs) and 35% natural gas. At fiscal year-end, approximately 92% of proved reserves were classified as proved developed producing and 8% as proved undeveloped.

Probable and Possible Reserves

The probable and possible reserves are categories that represent potential recoveries from the CO2 flood developed in the Delhi Field greater than those included in the proved reserves. Consequently, while the probable and possible reserves are 85% and 91% developed, respectively, and require de minimis capital expenditures, they remain less certain of attainment and have more risk of recovery than proved reserves and should not be aggregated with other categories. These categories of reserves reflect the incremental reserves associated with different engineering assumptions with respect to the percentage of original oil in place that can be recovered through CO2 enhanced oil recovery. Probable and possible reserves decreased approximately 1% and 8%, respectively, from the prior year primarily due to the movement of probable and possible reserves to the proved reserves category as a result of a positive change to the proved reserves life at Delhi

Investor Conference Presentation

Evolution today announced that the Company will participate in EnerCom's The Oil & Gas Conference in Denver, Colorado on August 17, 2021. The Company's presentation will be webcasted and is scheduled to be available at 11:40 am Eastern (10:40 a.m. Central) on August 17, 2021. The presentation can be accessed through the EnerCom conference portal for registered participants at https://www.theoilandgasconference.com, or in the investor relations section of the Company's website: http://www.EvolutionPetroleum.com.

About Evolution Petroleum

Evolution Petroleum Corporation is an oil and natural gas company focused on delivering a sustainable dividend yield to its shareholders through the ownership, management, and development of producing oil and natural gas properties onshore in the United States. The Company's long-term goal is to build a diversified portfolio of oil and natural gas assets primarily through acquisition, while seeking opportunities to maintain and increase production through selective development, production enhancement, and other exploitation efforts on its properties. Our largest assets are our interest in a CO2 enhanced oil recovery project in Louisiana's Delhi Field, our interest in a secondary recovery project in Wyoming's Hamilton Dome Field, and our recently acquired interest in the Barnett Shale in Texas. Additional information, including the Company's annual report on Form 10-K and its quarterly reports on Form 10-Q, is available on its website at http://www.EvolutionPetroleum.com.

Cautionary Statement

All forward-looking statements contained in this press release regarding current expectations, potential results and future plans and objectives of the Company involve a wide range of risks and uncertainties. Statements herein using words such as "believe," "expect," "plans," "outlook," "should," "will," and words of similar meaning are forward-looking statements. Although our expectations are based on business, engineering, geological, financial, and operating assumptions that we believe to be reasonable, many factors could cause actual results to differ materially from our expectations and we can give no assurance that our goals will be achieved. These factors and others are detailed under the heading "Risk Factors" and elsewhere in our periodic documents filed with the Securities and Exchange Commission. The Company undertakes no obligation to update any forward-looking statement.

Our reserves as of June 30, 2021 were estimated by DeGolyer & MacNaughton, a global independent reservoir engineering firm. All reserve estimates are continually subject to revisions based on production history, results of additional exploration and development, price changes and other factors. The SEC's current rules allow oil and gas companies to disclose not only proved reserves, but also probable and possible reserves that meet the SEC's definitions of such terms. Estimates of probable and possible reserves by their nature are much more speculative than estimates of proved reserves. These non-proved reserve categories are subject to greater uncertainties and the likelihood of recovering those reserves is subject to substantially greater risk. When estimating the amount of oil and natural gas liquids recoverable from a particular reservoir, probable reserves are those additional reserves that are less certain to be recovered than proved reserves but which, together with proved reserves, are as likely as not to be recovered, generally described as having a 50% probability of recovery. Possible reserves are even less certain and generally require only a 10% or greater probability of being recovered. These three reserve categories have not been adjusted to different levels of recovery risk among these categories and are therefore not comparable and are not meaningfully combined.

Company Contacts:

Jason Brown, President & CEOJBrown@evolutionpetroleum.com(713) 935-0122

Ryan Stash, SVP & CFORStash@evolutionpetroleum.com(713) 935-0122

SOURCE: Evolution Petroleum Corporation

View source version on accesswire.com: https://www.accesswire.com/659842/Acquisition-and-Positive-Revisions-Drive-Substantial-Increase-in-Evolution-Petroleums-Proved-Reserves-for-Fiscal-Year-End-2021

Go here to read the rest:

Acquisition and Positive Revisions Drive Substantial Increase in Evolution Petroleums Proved Reserves for Fiscal Year-End 2021 - Yahoo Finance

Restricting the evolutionary space of BL21(DE3) for the production of toxic proteins

In DE3 strains, the strong lacUV5 promoter drives expression of T7 RNAP, and it was previously shown that homologous recombination between lacUV5 and the weaker wild-type lac promoter is a dominating event that within hours of introducing a toxic gene leads to tolerance in BL21(DE3)15. To prevent this, we first genetically restricted BL21(DE3), creating a lacIlacZ variant by deleting exactly the part of the native lac locus that shared homology with the DE3 locus (Fig.1). In addition, this should prevent the expression of lacY, encoding the lactose permease, allowing a more uniform, concentration-dependent entry of IPTG into all cells in bacterial populations16.

a Illustration showing the experimental set-up used to restrict the evolutionary space for BL21(DE3) to overcome protein production toxicity on three levels: (upper panel) genetic restriction was accomplished by deleting a part of the BL21(DE3) genome that frequently recombines and lowers the T7 RNAP expression, (middle panel) protein production was coupled to both fluorescence and antibiotic resistance to prevent the formation of non-producing mutants and (lower panel) a spatiotemporally structured environment of ageing bacterial colonies was designed in order to identify strong phenotypic mutants. b Schematic of the workflow to produce layered agar plates that allow diffusion of the antibiotic ampicillin (AMP)and the inducer IPTG to allow sufficient time for colony formation on the surface of the top agar layer before toxic protein production is induced.

Next, similar to a previous study12, we chose the E. coli membrane protein insertase YidC coupled to GFP to serve as a model protein to investigate stress caused by membrane protein overproduction as it formerly was shown to have a strong negative fitness effect on E. coli17,18. The GFP fusion comes in handy for phenotypic restriction because it allows visual screening for mutants that still produce the fusion protein. We further restricted the evolutionary solution space by introducing a hairpin structure in the expression plasmid that couples YidC-GFP to the expression of a -lactamase gene19, providing resistance to ampicillin (Fig.1). This way, the formation of non-producing populations should be minimised in the presence of the antibiotic.

Finally, in contrast to the previous studies11,13,20, we aimed at introducing the protein production stress after bacterial colonies had established on plates. This spatiotemporally different approach should allow the formation of a large number of bacteria in a state of dormancy and in a structured environment, which previously was shown to constitute a unique evolutionary environment21. Furthermore, we speculated that mutants would be easy to identify as fluorescent secondary colonies, so-called papillae, outgrowing from the initially established colonies. To this end, two-layered agar plates were poured, allowing slow IPTG and ampicillin diffusion from the bottom to the top layer to grant sufficient time for colony formation before YidC-GFP production was induced (Fig.1).

During incubation for 1 week at 37C, we observed several fluorescent papillae (Fig.1) that were restreaked to confirm the fluorescent and ampicillin-resistant phenotype. Based on the fluorescence phenotype, the most promising mutants were isolated and cured of the YidC-GFP plasmid using a mild and simple CRISPR-based approach to plasmid curing22. These strains were subsequently retransformed with the original YidC-GFP plasmid to ensure that mutations leading to tolerance were localised on the genome and not on the expression plasmid. A single clone (Evo21(DE3)) was chosen for further characterisation.

To benchmark Evo21(DE3), we compared its ability to produce the YidC-GFP-fusion protein to the non-evolved BL21(DE3) wild-type strain as well as the derivative Mt56(DE3) previously described to be optimised for YidC-GFP overproduction12. The plasmid pLysS was utilised to limit basal T7 RNAP expression23 in BL21(DE3) and Evo21(DE3). Co-expression of pLysS in Mt56(DE3) is not relevant due to the greatly reduced polymerase activity in this strain. In liquid culture, based on GFP fluorescence, Evo21(DE3) expressed significantly (P0.0001) higher amounts of YidC-GFP than BL21(DE3) and Mt56(DE3). Eight hours post induction, yields were 3.6-fold higher in cultures of Evo21(DE3) than in cultures of BL21(DE3) and 2.1-fold higher than in Mt56(DE3) (Fig.2a). Parallel monitoring of the optical density of the cultures showed no major growth impairment for the strains (Supplementary Fig.1).

a Production of a toxic YidC-GFP-fusion protein in Evo21(DE3) compared to the non-evolved BL21(DE3) wild-type strain, the BL21(DE3)lacI/Z ancestor strain, and a previously evolved BL21(DE3) derivative Mt56(DE3). On the illustrated expression vector (pYidC), YidC-GFP production is translationally coupled to ampicillin resistance. b Fluorescence fold change between Evo21(DE3) and BL21(DE3), producing a library of 24 proteins of the E. coli inner membrane proteome C-terminally fused to GFP. c gfp expression levels in Evo21(DE3) and control strains with and without co-expression of the helper plasmid pLysS. d Western blot showing the expression of a camelid-derived single-chain antibody (nanobody) in Evo21(DE3) and control strains. Samples were normalised to cell density before loading. e Production of YidC under the control of a PrhaBAD promoter allowing titration of expression l-rhamnose. All fluorescence values displayed are normalised to OD at 600nm. Error bars indicated represent the average squared deviation from the mean (SD) of three biologically independent samples (n=3).

To assess whether the Evo21(DE3) phenotype was gene-specific, we next investigated the expression of a set of 24 different GFP-fusion proteins selected from an expression vector library of the E. coli inner membrane proteome24. These membrane proteins were selected to cover a wide range of functions, toxicity (previously reported as a change in OD600 upon IPTG addition24), and the number of predicted transmembrane domains (Supplementary Table1). Comparing the fold change of protein production in Evo21(DE3) and BL21(DE3), titre was improved for 19 of the 24 proteinswith a significant fold change of more than 1.5-fold for 14 of them (Fig.2b). The highest improvement was 6.1-fold (P0.001), observed for the protein YihG.

As a first test that the underlying mechanism allowing Evo21(DE3) to produce more toxic protein was not related to a general decrease in the activity of the T7 expression systemas previously observed for the BL21(DE3) derivatives C41/43(DE3), C44/45(DE3) and Mt56(DE3)we Sanger sequenced the T7 RNAP gene, which confirmed an absence of mutations. Next, we compared the expression of two non-toxic soluble proteins, GFP and a camelid-derived nanobody, and both were produced at higher levels in Evo21(DE3) than in BL21(DE3) or Mt56(DE3)both in the presence and absence of pLysS (Fig.2c, d). Similarly, Evo21(DE3) outperformed other strains in the production of seven out of eight different plant-derived cytochrome P450 enzymesa class of enzymes of significant biotechnological interest25 (Supplementary Fig.2). This indicates that the causative mutation in Evo21(DE3) is different from previously isolated BL21(DE3) derivatives and probably does not cause a general decrease in T7 RNAP activity.

Even though the screening for improved protein productivity was performed with the highly efficient T7 system, the ideal mutant strain would be capable of producing more protein independently from the promoter system. To explore if this was the case for Evo21(DE3), we replaced the T7 promoter with the l-rhamnose inducible rhaBAD promoter in the yidC-GFP expression vector, transformed it into Evo21(DE3), and expressed the construct by inducing with different rhamnose concentrations in liquid culture. With concentrations of 5 and 20mM l-rhamnose, Evo21(DE3) produced significantly (P0.0001) more protein than BL21(DE3) and Mt56(DE3) (Fig.2e).

In summary, this initial characterisation shows that the evolved strain can produce a higher titre of a range of different proteins using a T7-system-independent mechanism.

The phenotype of Evo21(DE3) prompted us to sequence the strain using Illumina whole-genome sequencing. Two point mutationsone in the argE and one in the fecB locus (Fig.3a)and an insertion of a mobile IS1 element into the rne gene were identified. Upon reintroduction of the argE or fecB point mutations into BL21(DE3) by oligonucleotide-based recombineering, the YidC-GFP overexpression phenotype was not obtained (data not shown), whereas, when reintroducing the truncation of the rne locus into BL21(DE3) and the Evo21(DE3) parental BL21(DE3) lacIlacZ strain, the YidC-GFP overexpression tolerance phenotype was nearly identical to Evo21(DE3) (Fig.3b). This makes it highly likely that the rne IS1 insertion is the main causative mutation in Evo21(DE3).

a Illustration of mutations in the Evo21(DE3) genome compared to the ancestor BL21(DE3). Whole-genome sequencing of mutant strain Evo21(DE3) revealed two point mutations (grey) and a truncation of the rne gene caused by the transposition of a mobile element IS1 into the locus. The deletion of the genomically shared homology sequence in the BL21(DE3) ancestor strain with the DE3 area (335,401337,123) is annotated. b Production levels of the toxic YidC-GFP-fusion protein in Evo21(DE3) compared to BL21(DE3), as well as BL21(DE3) and the non-evolved ancestor strain BL21(DE3)lacIlacZ after reintroducing the rne truncation by recombineering. Error bars indicated represent the average squared deviation from the mean (SD) of three biologically independent samples (n=3). c Illustration of the E. coli RNA degradosome. N- and C-terminal domain of the membrane-bound essential endonuclease RNase E (blue) and the localisation of associated enzymes PNPase, Rhlb and enolase along the C-terminal non-catalytic scaffolding region are displayed. Mutations of the rne gene in Evo21(DE3), BL21Star(DE3) and rne* gene harboured on pLysS-Max are indicated.

The identified IS1 insertion causes a truncation of the encoded 1061-residue E. coli endoribonuclease RNase E after amino acid 702 and, therefore, a polypeptide lacking the last 359 residues of its C-terminus in Evo21(DE3) (Fig.3c). RNase E is an essential membrane-associated enzyme involved in the maturation of both ribosomal RNA and tRNA, as well as total mRNA decay, and mediates the assembly of a multi-enzyme complex referred to as the RNA degradosome (Fig.3c). It has previously been shown that only the N-terminal half (1529) of RNase E, accommodating the active catalytic domain, is essential for cell growth, and the C-terminal non-catalytic region is mostly disordered and known to function as a scaffold mediating the association of the enzymes PNPase, Rhlb and enolase26,27,28.

Interestingly, a similar truncation of the rne locus, rne131, resulting in an RNase E polypeptide lacking its non-catalytic region (amino acid residues 1584, Fig.3c) was isolated in a screen for suppressors of a temperature-sensitive allele of the mukB gene26. A later study showed that in strains such as BL21(DE3), introducing the rne131 truncation caused a bulk stabilisation of mRNA degradation, including mRNA produced by T7 RNAP29. The rne131 truncation was engineered into the commercially available BL21Star(DE3) with the rationale that stabilising bulk mRNA would result in increased protein production. However, following the same rationale, the commercial strain also comes with a note suggesting that it might be unsuitable for overexpression of toxic genes.

We compared the expression of six different genes that we previously found expressed better in Evo21(DE3) than in BL21(DE3) with expression in BL21Star(DE3) and found expression levels to be highly similar between BL21Star(DE3) and Evo21(DE3) (Fig.4a). This provides an independent confirmation that the phenotype of Evo21(DE3) is caused by the truncation of rne.

a Heterologous production of a variety of non-toxic and toxic GFP-fusion proteins in Evo21(DE3) and BL21Star(DE3), both harbouring different truncations of the rne gene, compared to BL21(DE3). b Expression of the same genes in Evo21(DE3) is compared to expression in BL21(DE3) when co-expressing either the auxiliary plasmid pLysS or pLysS-Max. c Schematic illustration of the plasmid pRNE-GFP designed to report on RNase activity. Promoters controlling rne expression are indicated (P14)49,50. The expression level of the rne gene can be monitored in vivo via GFP fluorescence signal. d Exploration of the pRNE-GFP reporter plasmid in BL21(DE3) derivatives. Different levels of RNase activity can be monitored in the strains. e Titration of yidC expression in Evo21(DE3), BL21Star(DE3) and BL21(DE3) via increasing levels of l-rhamnose and its effect on the rne regulon expression reporting RNase activation in the cell during toxic membrane protein production. Upper half: Fluorescence corresponds to YidC-GFP production levels. Lower half: Cells harbour both the rne reporter plasmid (pRNE-GFP) and pPrhaBAD-YidC controlling yidC expression (no GFP fusion). Fluorescence levels correspond to GFP produced under the control of the rne regulon. f Effect of the pLyS-Max auxiliary plasmid on RNase activity. Plasmids pLysS and pLysS-Max are co-expressed along with pRNE-GFP in BL21(DE3) cells. To repress leaky expression of the rhamnose promoter controlling rne* expression on pLysS-Max, 0.4% glucose was added where indicated. Error bars indicated represent the average squared deviation from the mean (SD) of three biologically independent samples (n=3).

The way Evo21(DE3) was isolated from papillae outgrowing colonies on week-old agar plates, and because dominant rne mutants previously have been observed30, made us speculate that different rne variants could be studied by simple co-expression from a plasmid in the presence of wild-type rne on the genome. To test this idea and to compare different variants of the rne gene at different expression levels, we cloned rne variants in front of the rhaBAD promoter on the pLysS plasmid backbone: full-length rne, the Evo21(DE3) and BL21Star(DE3) truncated versions and a version with a further truncation in the membrane-binding domain of RNase E. However, none of these constructs showed any positive effect on YidC-GFP expression (Supplementary Fig.3).

Serendipitously, we isolated a spontaneously occurred rne mutant, called rne*, and included it in our analysis. We found that rne* provided on the pLysS plasmid (hereafter called pLysS-Max) could increase YidC-GFP production even further than Evo21(DE3) (Fig.4b). The rne* mutation converts the essential31 aspartate residue in position 346 to an asparagine in the so-called DNase I subdomain of RNase E involved in chelating an essential Mg2+ ion. The aspartate residue is believed to act as a general base to activate the attacking water essential for the catalytic activity of the enzyme32. The replacement of Asp-346 with the polar amino acid Asn was previously shown to decrease RNA cleavage by about 25-fold32. The effect of expressing rne* was not gene-specific as the effect was preserved for three out of four other tested genes (Fig.4b). This provides an alternative demonstration that manipulating with rne severely affects recombinant protein production and provides a simple tool, in the form of an auxiliary pLysS-Max plasmid that can be transformed into other strains, to improve protein production titre.

The positive effect of rne truncations such as rne131 on protein production was previously assumed to be due to the stabilisation of the recombinant mRNA29. However, the observation that the rne truncation in Evo21(DE3) leads to tolerance of toxic gene expression suggests a broader role involving balancing of RNA levels more globally.

Autoregulation allows RNase E to continuously adjust its synthesis to that of its substrates by controlling the degradation rate of its own mRNA33,34. This could work as a biosensor for RNase E activity by fusing the promoter and 5end of rne with a genetic reporter, as previously demonstrated with lacZ34. To explore RNase E activity in our evolved strains for recombinant protein production, we constructed a similar RNase E biosensor (pRNE-GFP) using GFP as a reporter (Fig.4c).

We transformed the pRNE-GFP reporter into BL21, BL21(DE3), BL21Star(DE3), Evo21(DE3) and Mt56(DE3) and monitored fluorescence in a microplate reader under conditions similar to the typical protein production scenario. Surprisingly, under these conditions, fluorescence was 14-fold reduced in Evo21(DE3) and sevenfold reduced in BL21Star(DE3) compared to the ancestral BL21(DE3 (Fig.4d). Given that Evo21(DE3) behaves almost identically to BL21Star(DE3) and that the rne131 truncation has been shown to cause a bulk stabilisation of mRNA degradation29, this suggests that low fluorescence from our reporter correlates with increased bulk mRNA stabilisation. Interestingly, fluorescence from the reporter was reduced approximately twofold in BL21 compared to BL21(DE3), suggesting an effect of the DE3 locus itself on RNase E activity in the cell.

Next, we wanted to explore if the expression of YidC-GFP affected RNase E activity in the different strains. To this end, because YidC-GFP cannot be expressed from the T7 promoter in BL21 (no T7 RNA polymerase) or BL21(DE3) (no growth), we expressed it from the rhaBAD promoter construct (Fig.2e) using different concentrations of rhamnose. This showed that YidC-GFP levels could be titrated with rhamnose and confirmed higher expression in BL21Star(DE3) and Evo21(DE3) than in the other strains at high rhamnose concentrations (Fig.4e, upper half).

To monitor fluorescence from the RNase E GFP reporter, we then deleted GFP from the YidC construct and attempted to co-transform it with pRNE-GFP into different strain backgrounds (Fig.4e, lower half). However, we were unable to recover and grow transformants in BL21(DE3) and BL21Star(DE3), suggesting a lethal imbalance in RNA levels caused by the presence of these two plasmids. We were able to recover and grow double transformants in BL21, Evo21(DE3), and Mt56(DE3) and monitor fluorescence as an indication of RNase E activity. Fluorescence increased to high levels upon increasing the concentration of rhamnose in BL21 and Mt56(DE3), but fluorescence levels were at least 2.5-fold lower (at 1mM rhamnose) in Evo21(DE3) and hardly increased upon further rhamnose addition. This suggests that RNase E activity towards bulk mRNA is increased when yidC expression is increased but that the activity is lower in Evo21(DE3) than in the other strains (Fig.4e).

Finally, we explored the effect of the pLysS-Max auxiliary plasmid (harbouring the mutant rne*) on RNase E activity by co-transforming it along with pRNE-GFP into BL21(DE3). As controls, we included a strain co-transformed with pLysS and pRNE-GFP and BL21(DE3) transformed only with pRNE-GFP. Because we previously observed effects on RNase E activity due to the presence of the DE3 locus, we repressed leaky expression of T7 RNAP from the lacUV5 promoter by adding glucose to the medium and titrated rne* levels with rhamnose. In the absence of glucose and the presence of pLysS, we observed an increase in fluorescence which was repressed by expression of rne* (Fig.4f). This shows that the pLysS-Max plasmid can be used to regulate RNase E activity in the cell.

Continue reading here:

Tailoring the evolution of BL21(DE3) uncovers a key role for RNA stability in gene expression toxicity | Communications Biology - Nature.com

What can we learn from a zebrafishs regenerating tail? Or a termite armys march into uncharted terrain? Welcome to the fields of ecology and evolution, where researchers observe the natural world to better understand how species are shaped by each other and their environments.

In practice, it takes years of notetaking and analysis to break down adaptations and other evolutionary forces. Charles Darwin noticed the uniquely shaped beaks of the Galapagos finches during a five-week foray to the islands, and then spent the next decade and a half trying to make sense of them.

But even a single moment can shed light on an organisms grind for survival, especially when its captured on camera. The BioMed Central Ecology and Evolution image competition highlights photos that show adaptations in action. Experts submit works from lab benches and field sites across the world to vy for the crowning spot in their area of study.

[Related: 14 hypnotizing photos that captured the world during the pandemic]

Here are the judges 2021 selections for each category, along with the grand prize winner.

See original here:

Animal photos that show the power of evolution - Popular Science

In 1988, Richard Lenski, a thirty-one-year-old biologist at UC Irvine, started an experiment. He divided a population of a common bacterium, E. coli, into twelve flasks. Each flask was kept at thirty-seven degrees Celsius, and contained an identical cocktail of water, glucose, and other nutrients. Each day, as the bacteria replicated, Lenski transferred several drops of each cocktail to a new flask, and every so often he stored samples away in a freezer. His goal was to understand the mechanics of evolution. How quickly, effectively, creatively, and consistently do microorganisms improve their reproductive fitness?

Lenskis flasks produced about six new generations of E. coli a day; the bacteria woke up as babies and went to bed as great-great-great-grandparents. In this way, Lenski and his team have studied more than seventy thousand generations of E. coli over thirty-three years. Compared with their distant ancestors, the latest versions of the bacterium reproduce seventy per cent faster; it once took them an hour to double their ranks, but now they can do it in less than forty minutes. Different populations have taken different paths to enhanced fitness, but, after decades, most have arrived at reproduction rates within a few percentage points of one another.

Lenskis Long-Term Evolution Experiment, or L.T.E.E., as its called, has yielded fundamental insights into the mutational capabilities of microorganisms. For his work, Lenski, now in his sixties and at Michigan State University, has received a MacArthur genius grant and a Guggenheim Fellowship. Im not sure I can tell you how its affected my thinking, because Im not sure I can conceive of being in this field without this experiment existing, Michael Baym, an evolutionary biologist at Harvard Medical School, recently told Discover.

Three of the experiments key findings are especially relevant today. The first is that, in general, there were diminishing returns to mutation over time: the bacteria made many of their most reproductively advantageous moves early on. A second finding, however, was that the bacteria never stopped getting fitter. Seventy thousand generations in, theyre still finding new ways to improve, albeit at a somewhat slower rate. I had sort of imagined that things would have flatlined, Lenski told me recently, when we spoke over Zoom. But there seem to be endless possibilities for tinkering and progress. If there is a hard limit, its so, so far away that its impractical to consider on an experimental timescalemaybe even a geological timescale.

Lenski has a friendly, expressive face, with pale blue eyes and a neat beard; his voice pulses with excitement when he considers a provocative question or explains the implications of his research. He told me about a third major finding: in 2003, some fifteen years and thirty thousand generations into the experiment, Lenski arrived at his lab to find that, overnight, a flask that was normally fairly translucent had turned cloudy. The bacteria it contained had experienced an explosive surge in growth. Normally, E. coli eat mainly glucose, but this population had unlocked an entirely new source of energy: a chemical compound called citrate. The capacity to metabolize citrate is so unusual that no population in the study had developed it until that point, and none have attained it since. Its as if a family of humans could suddenly drink salt water.

The emergence of a citrate-eating lineage suggests that exceptionally rare, profoundly consequential evolutionary leaps are possible long after the low-hanging fruit has been picked. How can something like this happen, and yet be so rare that it never happens again? Lenski asked. One possibility is that you have a super-rare mutation that is just so unique that its only happened once in the entire history of this experiment. Alternatively, maybe you need a whole set of earlier changes, so that you set up a genetic background where an ordinary mutation can allow this new function. I think its both: this was an unusual mutation, and it could only produce growth on citrate because there were specific genetic changes that preceded it.

SARS-CoV-2, the virus that causes COVID-19, has already had one citrate moment: the instant, probably sometime in 2019 but possibly earlier, when it developed the ability to leap into humans. Since then, the virus has accumulated innumerable mutations, some of which allow it to generate copies of itself more efficientlyby altering how it binds to our cells, for instance, or by finding new ways to slip past our immune systems. Its a process that has occurred with every infectious disease in historymeasles, tuberculosis, bubonic plague, influenza, and untold others. The difference with the coronavirus is that the world is now watching every mutational move as it happens.

During this pandemic, weve developed and deployed vaccines in real time. Meanwhile, SARS-CoV-2 is replicating not in a dozen flasks but in tens of millions of people, some of whom have been immunized, all of whom exert selective pressure for the virus to find new, more efficient replication strategies. The virus will continue to mutate every moment of every day, for years, for decades. The fear is that it will hit upon a second citrate moment: a mutation, or set of mutations, that enables it to circumvent our vaccines, which so far have proved spectacularly effective and resilient. For those who remain unvaccinatedthe majority of humankindthere is also the horrifying prospect of a variant that is vastly more contagious or deadly. Every few months, we learn of a version of the virus that seems somehow worse: Alpha, Beta, Gamma, Delta. The coronavirus appears destined to march its way through the Greek alphabeta prizefighter getting quicker, slicker, stronger with each opponent. What are the limits to its evolutionary fitness? Are they knowable? And, if so, how close are we to reaching them?

These were the questions on my mind as I spoke with experts in an effort to understand the future of the pandemic. With questions so complex, its helpful to start by figuring out what, exactly, we want to know. For each new coronavirus variant, we want to find out if its more transmissible, if it will make us sicker, and if it will more effectively get around our immune defenses. On that last front, we want to understand two more questions: How much will it succeed in hiding from our antibodies (which recognize and bind to the virus, preventing infection) and from our T cells (which recognize chopped-up viral fragments displayed by infected cells, and specialize not in preventing infection but in controlling and terminating it).

Roberto Burioni, a physician and professor at Vita-Salute San Raffaele University, in Milan, has been called the most famous virologist in Italy; he has written about the prospects for a final variant, a version of the coronavirus that has reached maximum transmissibility, and which becomes the dominant strain, experiencing only occasional, minimal variations. As Burioni sees it, there are three potential futures for the coronavirus. The firstthe most optimistic for usis one in which the virus simply cant evolve its way around the vaccines. This is not an unlikely possibility. Many virusesmeasles, mumps, rubella, polio, smallpoxhave never meaningfully circumvented their vaccines, and so far the best of our current jabs have remained remarkably protective against new coronavirus variants, including Delta.

A second possibility is that the virus will partially evade our vaccine-generated immune defenses while paying a price, becoming less infectious or lethal. In order for the coronavirus to hide from our antibodies, it has to change aspects of the key components recognized by our immune systems, including the spike protein; those changes could end up diminishing the proteins ability to bind to the receptors it needs to infect cells. We can consider, for example, the Beta and Gamma variants, which exhibit some level of immune evasion but havent become as infectious as Alpha or Delta. In the nineteen-nineties, H.I.V. experienced such a fate, when it hit upon a mutation known as M184V, which conferred resistance to the antiviral drug lamivudine. On the surface, this was a setback, but doctors soon learned that patients with the M184V variant had lower viral loads, suggesting that the mutation also reduced how efficiently the virus replicated inside the body. It became common for patients with H.I.V. to continue taking lamivudine even after resistance emerged, in part to select for the variant with a lower replication rate.

The third future is the most concerning: the virus could accumulate mutations that allow it to circumvent immunity without suffering a major reduction in transmissibility or lethality. This would require it to open up a new evolutionary spacea citrate moment. Even in this scenario, Burioni told me, were in a fortunate position: we can quickly modify our vaccines to confront new variants. At the same time, the manufacturing and distribution challenges facing those variant-specific boosters would be colossal; were struggling to fully vaccinate even a quarter of the worlds population with the vaccines we already have.

Vaccination is the most fundamental difference between Lenskis experiment and our reality. In Lenskis flasks, the environment is held constant; in the pandemic, we are doing everything we can to change it. A virus evolves one set of weapons when the world is immune-nave, and others as parts of the global population become fully vaccinated, partially vaccinated, andif immunity wanesformerly vaccinated. Different traits become more or less important in different settings. If youre a tennis fan, you might bet on Nadal on clay, Federer on grass, and Djokovic on a hard court. The question of whether a virus has reached something like peak fitness is inescapably linked to where, when, and whom its playing.

There is such a thing as peak fitness within a particular landscape, Kristian Andersen, an infectious-disease researcher at the Scripps Research Institute, told me. The problem is that the landscape keeps changing. That puts very strong selective pressure on the virus. The Beta and Gamma variants evolved in areas with high levels of prior infection, and so settled on mutations that offered them gains in immune escape but not transmissibility. The Delta variant, by contrast, emerged in India, which had relatively low vaccination levels; its goal was to spread as fast and as far as possible. Although it may be somewhat more immune-evasive and lethal, Deltas defining feature is its extreme contagiousness.

Some scientists argue that there are strict limits on how effectively the virus can slip past our immune defenses while retaining its infectiousness. Our antibodies recognize the specific parts of the virus that it uses to enter cells; the virus may alter these, but in doing so it may become a less effective invader. There are certainly limits, Andersen told me. We just have no idea where they are. The fundamental question is: How tolerant is the virus to these mutations? Can it still do what it needs to donamely, enter the cellwith a substantially mutated spike protein? Coronaviruses are generalists: they can bind to ACE-2 receptors, their ports of entry into cells, in many ways, across many species. We often use a lock-and-key model to understand how a protein binds to a receptor, Andersen said. That doesnt tell the whole story here: coronaviruses have shown they have many keys that can open the same lock.

Tyler Starr, an evolutionary biologist at the Fred Hutchinson Cancer Research Center, shares Andersens concerns. Starr recently led an ambitious project mapping all possible mutations to a key part of the spike protein. He wanted to know how the proteins structureand, therefore, its affinity for ACE-2changes as each amino acid in its receptor-binding sites mutated. The findings were not terribly reassuring. The big picture is that there is not that much biological constraint, Starr said. Theres a ton of tolerated mutational space that the virus can take while trying to evade immunity. Its quite flexible. Some researchers have seen as good news the fact that many variants share similar mutations despite having evolved separatelya phenomenon known as convergent evolution. According to one view, this means that the virus has a limited toolbox from which to work. According to another, these are only the easiest and earliest mutational options; future variants could hit upon more innovative ways to enhance transmissibility and evade immune defenses. The situation is further complicated by the fact that, unlike in Starrs experiment, the real-world virus isnt limited to one change at a time: it can combine multiple mutations to vastly expand its evolutionary space.

Still, there is reason for optimism. As James Somers explained in this magazine, last year, the human immune system is staggeringly complex and, over millennia, has honed countless defenses against microscopic intruders. And as Katherine Xue wrote, last month, it is especially effective when it has previously encountered a pathogen. In 2009, when the H1N1 influenza strain emerged, it had a curious feature: it caused more severe illness in younger people than older people. Globally, four in five H1N1 deaths were estimated to have occurred in people under sixty-five. (Typically, some seventy to ninety per cent of flu deaths occur in older adults.) It turned out that many older people had likely been exposed to a relative of the strain decades agoand that their immune systems, remembering that fight, were prepared for the next one.

Regardless of how drastically they mutate, new coronavirus variants will probably have more in common with the original SARS-CoV-2 than with SARS-CoV-1, the virus that caused the SARS outbreak in 2003. Even so, the blood of COVID-19 survivors has the potential to neutralize the 2003 strain. The vaccines from Pfizer and Moderna, similarly, appear to generate huge numbers of antibodies that work against SARS-CoV-1 in those who have also been infected with COVID-19. These two viruses span a really large evolutionary distance, Starr told me. The fact that the same antibodies bind to both of them should give us some confidence. With new coronavirus variants, we may see a partial decrease in immunity, but, given the polyclonal response, Starr saidthe fact that vaccines generate not one type of antibody but manywhen one set of antibodies drops the rope, another will pick it up. I dont think there will ever be a variant that completely escapes our immune systems. Were never going to wipe the slate clean and be back to a totally nave population. Over time, the infections we do get will be more likely to be mild or asymptomatic. Whether that process takes a year, five years, ten years, or longer, I dont know.

Continued here:

The New Yorker How Will the Coronavirus Evolve? - The New Yorker

Overall Winner and Best Image for Conservation Biology. A school of jack fish in a spiral formation at Heron Island in the Great Barrier Reef. A visual metaphor for the spiraling crisis unfolding within our oceans and the need for concentrated efforts to protect marine ecosystems. Credit: Kristen Brown

From furry crustaceans to hunting wasps and escaping frogs, the 2021BMC Ecology and EvolutionImage Competition has produced an impressive collection of celebrated images that showcase the diversity of Earths animal and plant life. All images are open access and available for use under a Creative Commons Attribution 4.0 (CCBY) license.

The overall winning image by Kristen Brown from the University of Pennsylvania, Philadelphia, USA depicts a school of jack fish in a spiral formation at Heron Island in the Great Barrier Reef, Australia.

Runner Up and Best Image for Evolutionary Developmental Biology and Biodiversity. Eulimnogammarus verrucosus, a species of crustacean endemic to the UNESCO World Heritage Site Lake Baikal, suffering from a parasitic ciliate infection. Credit: Kseniya Vereshchagina

Kristen Brown said: This image represents both the beauty and bounty of our oceans as well as the spiraling crisis unfolding within the marine environment. Coral reefs with high coral cover and plentiful fish populations like this one at Heron Island on the Great Barrier Reef are sadly becoming rarer. Without a concentrated effort to reduce greenhouse gas emissions and improve water quality, coral reefs as we know them are at risk of disappearing within our lifetime.

Best Image for Behavioural Ecology. The Hunter depicts a wasp and its spider prey in Tiputini, Ecuador. Credit: Roberto Garca-Roa

Section editor Josef Settele recommended the entry, saying: Marine biodiversity sustains life and the health of our planet, but human activities are threatening the well-being of the worlds oceans. Kristen Browns striking image is a symbol for the need for concentrated efforts to manage biodiversity loss and set conservation priorities.

In addition to the winning image, the judges also selected an overall runner up, as well as winners in six categories: Conservation Biology; Evolutionary Developmental Biology and Biodiversity; Behavioural Ecology; Human Evolution and Ecology; Ecological Developmental Biology; Population Ecology; and the Editors Pick. The winning images celebrate Earths biodiversity and its evolutionary origins, from how species learn and develop, to conflict, collaboration and parasitic relationships, both between and within species.

Best Image for Population Ecology. Small Big Migration captures a moment in the life of a population of soldier termites as they migrate to ensure survivorship and reproduction of the colony. Credit: Roberto Garca-Roa

The Population Ecology category winner was captured by Roberto Garca-Roa from University of Valencia, Spain, who also submitted the winning images for the Behavioural Ecology and Human Evolution and Ecology categories. It shows soldier termites migrating along a length of abandoned rope in a Malaysian forest.

Roberto Garca-Roa said: Thousands of soldier termites are able to migrate in a complex social environment where each individual has its own mission framed altogether in a global objective: the survivorship and reproduction of the colony. In this case, these termites used meters of an abandoned rope to move across the Malaysian forest. Once humans disappear, nature recovers its space and uses what is needed to survive.

Best Image for Human Evolution and Ecology. Learning to Be Human captures a researcher using a baboon to study the evolution of human locomotion. Credit: Roberto Garca-Roa

The Editors pick titled Eerie Stalker by Dimitri Ouboter from the Institute for Neotropical Wildlife and Environmental Studies, Suriname captures a Giant Gladiator Frog seconds before escaping from an attempted snake attack. Giant Gladiator Frogs have been previously observed escaping from the jaws of snakes by emitting distress calls, jumping and inflating their lungs, making it harder for small snakes to hold on to them.

Best Image for Ecological Developmental Biology. A zebrafish regrew its tail fin only two weeks after the appendage was clipped at the white horizontal dotted line. Credit: Chey Chapman

TheBMC Ecology and EvolutionImage Competition was created to give ecologists and evolutionary biologists the opportunity to use their creativity to highlight their work and celebrate the intersection between art and science. It follows on from theBMC Ecologycompetition, which ran for seven years untilBMC Ecologymerged withBMC Evolutionary Biologyto formBMC Ecology and Evolution. The winning images are selected by the Editor ofBMCEcology and Evolutionand senior members of the journals editorial board.

Editors pick. Eerie Stalker depicts a giant gladiator frogs escape from a snake. Credit: Dimitri Ouboter

Editor Jennifer Harman said: We had a wonderful experience judging the fantastic images submitted to this years competition. Our section editors used their expertise to ensure the winning images were picked as much for the scientific stories behind them as for the technical quality and beauty of the images themselves. As such, the competition very much reflects BMCs ethos of innovation, curiosity and integrity. We thank all those who took part in this years competition; we hope that our readers enjoy viewing these images and discovering the stories behind them.

Reference: Inaugural BMC Ecology and Evolution image competition: the winning images by Jennifer L. Harman, Alison L. Cuff, Josef Settele, Luke M. Jacobus, David A. Liberles and Arne Traulsen, 12 August 2021, BMC Ecology and Evolution.DOI: 10.1186/s12862-021-01886-7

Read the original:

2021 BMC Ecology and Evolution Image Competition: See the Spectacular Winning Photographs - SciTechDaily

DUBLIN--(BUSINESS WIRE)--The "Nonresidential Prefabricated Building Systems - Global Market Trajectory & Analytics" report has been added to ResearchAndMarkets.com's offering.

Global Nonresidential Prefabricated Building Systems Market to Reach $94 Billion by 2027

Amid the COVID-19 crisis, the global market for Nonresidential Prefabricated Building Systems estimated at US$67.9 Billion in the year 2020, is projected to reach a revised size of US$94 Billion by 2027, growing at a CAGR of 4.8% over the analysis period 2020-2027.

Metal Building Systems, one of the segments analyzed in the report, is projected to record a 4.6% CAGR and reach US$47.7 Billion by the end of the analysis period. After an early analysis of the business implications of the pandemic and its induced economic crisis, growth in the Modular Building Systems segment is readjusted to a revised 5.4% CAGR for the next 7-year period.

The U.S. Market is Estimated at $20 Billion, While China is Forecast to Grow at 4.5% CAGR

The Nonresidential Prefabricated Building Systems market in the U.S. is estimated at US$20 Billion in the year 2020. China, the world's second largest economy, is forecast to reach a projected market size of US$16.6 Billion by the year 2027 trailing a CAGR of 4.5% over the analysis period 2020 to 2027.

Among the other noteworthy geographic markets are Japan and Canada, each forecast to grow at 4.5% and 3.8% respectively over the 2020-2027 period. Within Europe, Germany is forecast to grow at approximately 4% CAGR.

Panelized Precast Concrete Systems Segment to Record 4.7% CAGR

In the global Panelized Precast Concrete Systems segment, USA, Canada, Japan, China and Europe will drive the 4.7% CAGR estimated for this segment. These regional markets accounting for a combined market size of US$9.9 Billion in the year 2020 will reach a projected size of US$13.6 Billion by the close of the analysis period.

China will remain among the fastest growing in this cluster of regional markets. Led by countries such as Australia, India, and South Korea, the market in Asia-Pacific is forecast to reach US$10.8 Billion by the year 2027.

Key Topics Covered:

I. METHODOLOGY

II. EXECUTIVE SUMMARY

1. MARKET OVERVIEW

2. FOCUS ON SELECT PLAYERS (Total 211 Featured):

3. MARKET TRENDS & DRIVERS

4. GLOBAL MARKET PERSPECTIVE

III. REGIONAL MARKET ANALYSIS

IV. COMPETITION

For more information about this report visit https://www.researchandmarkets.com/r/ji7re4

Go here to see the original:

Global Nonresidential Prefabricated Building Systems Market Report 2021: Evolution of Prefabricated Buildings as the Revolutionary Change in the...

During a drill in the early portion of Arkansas State's practice Wednesday, an assistant coach reminded the Red Wolves to stay on their feet.

Although they had helmets and shoulder pads on, it was just a thud session -- hitting without going to the ground.

That won't be the case this afternoon when Arkansas State University has its first scrimmage of fall camp on the turf inside Centennial Bank Stadium. Just more than a week into the first preseason camp of the Butch Jones era, it'll be the first opportunity for the Red Wolves' coaches to evaluate their defense in a gamelike scenario.

"It's a critical day in the evolution and the evaluation of our football team," Jones said earlier this week. "It's an opportunity for them to really put their identity on video. We're finding out the players who we can win with, who we can trust in critical situations, and really just let them go out and play football."

Although many of the names who finished last season as defensive starters are back for Arkansas State, defensive coordinator Rob Harley is in the process of transitioning the Red Wolves from a 3-4 to a 4-3 base defense.

The only player who appears to have cemented himself on defense is senior linebacker Caleb Bonner, who Jones repeatedly has praised for his leadership.

It means there's ample opportunity for transfers such as Kivon Bennett, John Mincey and Joe Ozougwu to assert themselves along the defensive line. Arkansas State ranked 115th out of 127 teams last year in yards allowed per game, and the Red Wolves weren't helped by a pass rush that Pro Football Focus graded as 114th among all FBS teams.

They'll all get a chance in a situational scrimmage that Jones said will feature the first-team offense going against the first-team defense. The plan is to include live special teams while focusing on all types of scenarios -- red zone, move the chains and backed-up deep in the offense's territory.

Based on practice, Layne Hatcher will run the first-team offense while Florida State transfer James Blackman will man the second unit.

But Hatcher could have at least half of his group different from the one with which he finished 2020. TCU transfer Te'Vailance Hunt slotted in at wide receiver opposite Jeff Foreman and tight end Reed Tyler, and the offensive tackle spots were manned by Kentucky transfer Nick Lewis and Austin Peay transfer Robert Holmes during the open portion of Wednesday's session

That doesn't include redshirt freshman Ethan Miner, who was at center alongside returners Andre Harris and Ivory Scott, and Corey Rucker, who was limited earlier in the week after suffering a shoulder injury but should be close to a full participant today.

"The big thing is just focusing one day at a time and continuing to evaluate and see where we're at," Jones said. "Everything is evaluated in our program -- every single rep, whether it's a team rep, an individual rep, leadership reps, accountability reps. All those things in our program are evaluated on a daily basis."

Senior linebacker Caleb Bonner (center) is in a leadership role for a Red Wolves defense thats intent on improving this season. Arkansas State will have its first scrimmage of fall camp today at Centennial Bank Stadium in Jonesboro.(Photo courtesy Arkansas State Athletics)

Arkansas State wide receiver Jeff Foreman caught 13 passes for 439 yards and 3 touchdowns last season. Foreman will take part in the Red Wolves first scrimmage of fall camp today at Centennial Bank Stadium in Jonesboro.(Courtesy Arkansas State Athletics)

Originally posted here:

ASU's 'evolution' takes another step - Arkansas Online

eFootball is a fresh start for Konamis long-running Pro Evolution Soccer (PES) series. Not only does the game take a radically different approach by switching from a paid annual release to a free-to-play model, but its also being built on an entirely new game engine this time around: Unreal Engine 4.

The shift has naturally led to questions over whether eFootball will be able to overcome some of the typical pitfalls and challenges that the free-to-play model can pose, and how content will be delivered to players after release.

In an exclusive interview with TechRadar, we spoke with eFootballs producer Seitaro Kimura about how the development team will manage cross-play, the benefits of working with Unreal Engine 4, and what he considers eFootballs biggest innovation to be.

From Pro Evolution Soccer to PES, and now eFootball Kimura-san explained that the decision to scrap the series long-running name (known as Winning Eleven in Japan) was down to a multitude of factors. Chiefly, the prospect of turning the game into a viable esport.

We were working on a project to revamp the PES game engine to coincide with the release of the next-generation of home game console such as the PlayStation 5 and Xbox Series X|S, says Kimura-san.

At the same time, we knew that we needed to meet the needs of the market by making the game free-to-play and cross-platform, allowing players to compete and cooperate across devices.

Following the major changes in the game engine and business aspects, we [wanted to] create a large cross-platform esports scene. To this end, we made the decision to unify Winning Eleven in Japan and PES overseas into eFootball.

eFootball is powered by Unreal Engine 4, replacing Konamis FOX Engine that powered the likes of Metal Gear Solid V: The Phantom Pain and previous iterations of PES behind. Building a game in an entirely new engine is always a challenge, but Kimura-san that Unreals development community helped make the series transition to Unreal Engine 4 a little smoother.

With the Unreal Engine, there are many more options for approaches, and with so many people using the Unreal Engine, I can refer to a lot of knowledge, says Kimura-san. Also, at UNREAL FEST, I was able to listen to development case studies of titles that had adopted Unreal Engine, so I was able to get more practical information.

Also, because we were able to develop for all platforms in one environment, we were able to handle cross-platform play relatively smoothly. The gameplay portion of the game uses an in-house football engine, so the integration was very challenging.

eFootball is set to launch with nine officially licensed teams in total, and we now know that new teams will be added and updated for free, along with individual squads. We will also continue to offer Live Update, a free data update service, so each teams transfers and squad changes will be updated weekly, says Kimura-san.

There will also be the predictable mix of licensed teams and made up ones, according to Kimura-san. There will only be a small number of teams available at launch, but other licensed teams with made up names will be available as usual via free updates.

Popular offline modes such as Master League, will be available in the future as premium downloadable content in eFootball, and the game will feature a Match Pass system that allows you to pick your favorite players and earn items based on the numbers of matches you play. Both free and paid passes will be available, says Kimura-san. Paid elements will include individual players for our new Team Building Mode, the aforementioned Match Passes, and we plan to sell additional modes in the future."

But how will eFootball balance the multiplayer and single-player components when it comes to fixes and future updates, and avoid falling into the Pay to win trap that so many free-to-play games succumb to?

We do plan to sell the existing offline modes as premium content in the future. As for those fixes and updates, they will be done in the same way for all modes, says Kimura-san. We are focusing on making sure that all players can enjoy a fair game, so the game specifications will not be a Pay to Win system where paid elements will determine who wins or loses in a match.

Unlike FIFA 22, eFootball will let you play with friends no matter which platform theyre on, including mobile. If you do opt for the mobile version, which will also support controllers in a future update, the good news is you wont always need a Wi-Fi connection if you want to get your football fix on, Kimura-san confirms.

Yes, it is possible to play online using 4G or 5G connection, Kimura-san explains. We are developing and adjusting so that differences in communication quality depending on the environment you are playing in will have as little impact on gameplay as possible.

But will there be a Nintendo Switch version of eFootball? Perhaps, though Kimura-san failed to provide an answer at this point. He did shed some light on what PS5 and Xbox Series X players can expect when they boot up eFootball for the first time, however.

Please expect to see major visual enhancements on PS5 and Xbox Series X, says Kimura-san. However, there is no 120Hz support on console.

While switching to free-to-play and choosing a new name is certainly a big deal for eFootball, its success will ultimately be decided by how it recreates the beautiful game. And when it comes to improvements on the pitch, Kimura-san believes that by focusing on the 1v1 element of football, adding a completely new animation system, and by working with some of the best players in the world, eFootball will ultimately benefit.

The most interesting part of football, in my opinion, is the 1v1 attack and defense. The 1 vs. 1 gameplay is the most innovative aspect of eFootball and will change the balance of the game, says Kimura-san.

In order to make this happen, we have innovated in every way possible, Kimura-san explains. In order to understand how the best players in the world play, we brought in [Andreas] Iniesta and [Gerard] Piqu as our gameplay advisors and asked them for their advice.

In addition, realistic gameplay requires both varied animations and high responsiveness. To achieve this, we built a completely new animation system, including technology we call Motion Matching. You can enjoy the delicate touch of the ball, feinting to deceive your opponent, defending with your body, or blocking a shot at the last second.

Interestingly, this new approach has led to eFootball adopting a control scheme similar to FIFA, with the triggers now controlling sprint and close control instead of the shoulder buttons.

With the revamp of the game engine, we have focused on the most exciting part of football, the 1v1 battle between players. As we created the game system by deconstructing the mechanics of winning and losing in real-life 1v1 battles and the tactics used by top-level players, we had to restructure the control method in order for players to enjoy the evolved tactics, says Kimura-san. Also, instead of having more complicated controls, the game is more intuitive and allows players to focus on the gameplay against their opponents, so we believe that people who have enjoyed the PES series in the past will be able to get used to it and enjoy playing against each other.

We wont have to wait long until we can go hands-on with eFootball, which will release early Autumn (likely September) for PS4, PS5, Xbox One, Xbox Series X/S, PC and mobile. Whether the game will be a championship contender or relegation fodder remains to be seen.

Read the original here:

How the (pro) evolution of Konamis eFootball came to be - TechRadar