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Abstract The deep continental crust represents a vast potential habitat for microbial life where its activity remains poorly constrained. Organic acids like acetate are common in these ecosystems, but their role in the subsurface carbon cycle - including the mechanism and rate of their turnover - is still unclear. Here, we develop an isotope-exchange ‘clock’ based on the abiotic equilibration of H-isotopes between acetate and water, which can be used to define the maximum in situ acetate residence time. We apply this technique to the fracture fluids in Birchtree and Kidd Creek mines within the Canadian Precambrian crust. At both sites, we find that acetate residence times are

Researchers point to several obstacles in higher education that can impact student veterans. For some, learning to seek help when needed is often a challenge. Student veterans may feel they must live up to a false expectation and not burden others with their problems, and have indicated challenges with interactions with faculty and difficulty with disability accommodations. Other researchers suggest that student veterans struggle with being more mature, being impatient with their traditional-age civilian student peers, and having a student life that is less structured than military life. Texas State University has had success with creating and implementing a learning frameworks course for student veterans and military-connected students. Course evaluations and feedback have been extremely positive as student veterans have indicated they have become more self-directed learners with the tools to persist in college, doing so in a way that is focused on their needs inside and outside the classroom, and both on and off-campus.

Since the 1920s, colleges have offered how-to-study courses orientating students to the rituals of academic study. We present an historical overview of these courses by highlighting major developments including the emergence of courses that evolved from skill-based curriculum underpinned by behaviorism to strategy-based curriculum underpinned by cognitive psychology and self-regulation theories. We also focus on two unique common iterations of course categories offered today: first-year seminars, which were re-envisioned by the University of South Carolina in the 1970s, and theory-based learning strategy courses, often referred to as learning frameworks courses, which emerged in the 1970s at two Texas universities.

Abstract Bacteroidales (syn. Bacteroidetes) are prominent members of the human gastrointestinal ecosystem mainly due to their efficient glycan-degrading machinery, organized into gene clusters known as polysaccharide utilization loci (PULs). A single PUL was reported for catabolism of high-mannose (HM) N-glycan glyco-polypeptides in the gut symbiont Bacteroides thetaiotaomicron, encoding a surface endo-β-N-acetylglucosaminidase (ENGase), BT3987. Here, we discover an ENGase from the GH18 family in B. thetaiotaomicron, BT1285, encoded in a distinct PUL with its own repertoire of proteins for catabolism of the same HM N-glycan substrate as that of BT3987. We employ X-ray crystallography, electron microscopy, mass spectrometry-based activity measurements, alanine scanning mutagenesis and a broad range of biophysical methods to comprehensively define the molecular mechanism by which BT1285 recognizes and hydrolyzes HM N-glycans, revealing that the stabilities and activities of BT1285 and BT3987 were optimal in markedly different conditions. BT1285 exhibits significantly higher affinity and faster hydrolysis of poorly accessible HM N-glycans than does BT3987. We also find that two HM-processing endoglycosidases from the human gut-resident Alistipes finegoldii display condition-specific functional properties. Altogether, our data suggest that human gut microbes employ evolutionary strategies to express distinct ENGases in order to optimally metabolize the same N-glycan substrate in the gastroinstestinal tract.

Abstract Carbon and chlorine isotope effects for biotransformation of chloroform by different microbes show significant variability. Reductive dehalogenases (RDase) enzymes contain different cobamides, affecting substrate preferences, growth yields, and dechlorination rates and extent. We investigate the role of cobamide type on carbon and chlorine isotopic signals observed during reductive dechlorination of chloroform by the RDase CfrA. Microcosm experiments with two subcultures of a Dehalobacter‐containing culture expressing CfrA—one with exogenous cobamide (Vitamin B12, B12+) and one without (to drive native cobamide production)—resulted in a markedly smaller carbon isotope enrichment factor (εC, bulk) for B12− (−22.1 ± 1.9‰) compared to B12+ (−26.8 ± 3.2‰). Both cultures exhibited significant chlorine isotope fractionation, and although a lower εCl, bulk was observed for B12− (−6.17 ± 0.72‰) compared to B12+ (−6.86 ± 0.77‰) cultures, these values are not statistically different. Importantly, dual‐isotope plots produced identical slopes of ΛCl/C (ΛCl/C, B12+ = 3.41 ± 0.15, ΛCl/C, B12− = 3.39 ± 0.15), suggesting the same reaction mechanism is involved in both experiments, independent of the lower cobamide bases. A nonisotopically fractionating masking effect may explain the smaller fractionations observed for the B12− containing culture.

The article provides recent state and system wide developmental education (DE) policy mandates and reforms for all 50 states. The rationale is to determine how states are refining the assessment, placement, instruction, and advisement of students deemed noncollege ready and who are advised or mandated to enroll in traditional or innovative DE instruction. This research extends previous DE policy research published from Education Commission of the States and other policy analysis organizations, research centers, foundations, and independent researchers. We used purposeful sampling to collect data from multiple sources (e.g. legislative statutes and mandates, state or system wide policy documents, agency records, etc.). Basic findings include 42 states that address, in some form, DE policy; 38 states that address DE assessment policy; 37 states that address DE course placement policy; 32 states that address DE instructional reform policy; and 29 states that address advising policy, in which 18 of those state policies are specific to students enrolled in DE. We also present a policy snapshot for each state. Trends from the research include standardized assessments being widely implemented with many states allowing for multiple measures of assessment as well as advising to configure placement into DE. Corequisite models of instruction are also becoming more common as an alternative to prerequisite DE stand-alone courses scaffolded by learning support. We posit that DE policies are emergent and mostly in continual flux and recommend the field would benefit from a common and consistent set of DE data for all 50 states.

The Office of Correctional Education was created through an Act of Congress in 1991 to oversee and coordinate prison education programs as a way to reduce recidivism (Carl D. Perkins Vocational and Applied Technology Education Act, 1990). However, correctional education completion rates are extremely low. Therefore, we used secondary data from the Program for the International Assessment of Adult Competencies survey to show that students in prison would benefit from developmental and student supports. Survey data represented 1,319 prisoners and 8,670 from the household population. We used descriptive analysis, chi-square tests, and Welch's t-tests to analyze survey data. We conclude that those in the prison sample had a greater potential need for student support programs than the household sample and would benefit from increased developmental and student supports.

"Fungi on Mars!": a popular news heading that piques public interest and makes scientists' blood boil. While such a statement is laden with misinformation and light on evidence, the search for past and present extraterrestrial life is an ongoing scientific effort. Moreover, it is one that is increasingly gaining momentum with the recent collection of martian rock cores from Jezero Crater by NASA's Perseverance rover. Despite the increasingly sophisticated approaches guiding the search for microbial life on other planets, fungi remain relatively underexplored compared to their bacterial counterparts, highlighting a gap between the astrobiological and fungal research communities. Through a meeting in April 2021, the CIFAR Earth 4D and Fungal Kingdom research programs worked to bridge this divide by uniting experts in each field. CIFAR is a Canadian-based global research organization that convenes researchers across disciplines to address important questions facing science and humanity. The CIFAR Earth 4D: Subsurface Science & Exploration and Fungal Kingdom: Threats & Opportunities research programs were launched by CIFAR in July 2019, each made up of approximately two dozen international researchers who are experts in their fields. The Earth 4D program, led by co-directors John Mustard (Brown University, USA) and Barbara Sherwood Lollar (University of Toronto, Canada), aims to understand the complex chemical, physical, and biological interactions that occur within and between Earth's surface and subsurface to explore questions on the evolution of planets and life. The Fungal Kingdom program, led by co-directors Leah Cowen (University of Toronto, Canada) and Joseph Heitman (Duke University, USA), seeks to tackle the most pressing threats fungi pose to human health, agriculture, and biodiversity and to harness their extraordinary potential. The programs met to explore areas for synergy within four major themes: (1) the origins of life; (2) the evolution and diversification of life; (3) life in diverse and extreme environments; and (4) extinction: lessons learned and threats. This report covers the research discussed during the meeting across these four themes.

Abstract Investigations of abiotic and biotic contributions to dissolved organic carbon (DOC) are required to constrain microbial habitability in continental subsurface fluids. Here we investigate a large (101–283 mg C/L) DOC pool in an ancient (>1Ga), high temperature (45–55 °C), low biomass (102−104 cells/mL), and deep (3.2 km) brine from an uranium-enriched South African gold mine. Excitation-emission matrices (EEMs), negative electrospray ionization (–ESI) 21 tesla Fourier-transform ion cyclotron resonance mass spectrometry (FT-ICR MS), and amino acid analyses suggest the brine DOC is primarily radiolytically oxidized kerogen-rich shales or reefs, methane and ethane, with trace amounts of C3–C6 hydrocarbons and organic sulfides. δ2H and δ13C of C1–C3 hydrocarbons are consistent with abiotic origins. These findings suggest water-rock processes control redox and C cycling, helping support a meagre, slow biosphere over geologic time. A radiolytic-driven, habitable brine may signal similar settings are good targets in the search for life beyond Earth.

Abstract The large volume of deep groundwater in the Precambrian crust has only recently been understood to be relatively hydrogeologically isolated from the rest of the hydrologic cycle. The paucity of permeability measurements in Precambrian crust below 1.3 km is a barrier to modeling fluid flow and solute transport in these low porosity and permeability deep environments. Whether permeability-depth relationships derived from measurements shallower than 1.3 km can be extended to greater depths in unclear. Similarly, application of a widely-used permeability-depth relationship from prograde metamorphic and geothermal systems to deep Precambrian rocks may not be appropriate. Here, we constrain permeabilities for Precambrian crust to depths of 3.3 km based on fluid residence times estimated from noble gas analyses. Our analysis shows no statistically significant relationship between permeability and depth where only samples below 1 km are considered, challenging previous assumptions of exponential decay. Additionally, we show that estimated permeabilities at depths >1 km are at least an order of magnitude lower than some previous estimates and possibly much lower. As a consequence, water and solute fluxes at these depths will be extremely limited, imposing important controls on elemental cycling, distribution of subsurface microbial life and connections with the near-surface water cycle.

Service-learning integrates community service with academic study. Students apply skills and knowledge in practical situations and reflect on their experiences. Our 3000-level service-learning college course, Leadership Development in Agricultural Sciences, involved student-led development and execution of a one-day event, “AgCamp,” held on a university campus for high school students. The objective of this study was to evaluate if an intervention (i.e., a service-learning project) changed undergraduate students’ perceptions of their ability to collaborate, ability to lead, and overall perceptions of the usefulness of service-learning. An electronic mixed methods questionnaire-based survey was developed, tested for validity and reliability, and distributed to college students enrolled in Leadership Development in Agricultural Sciences as a pre- and post-test at the beginning and end of two semesters. Data were analyzed in SPSS 26.0 using measures of central tendency and paired sample t-tests. Undergraduate students’ (n = 74) had positive perceptions of service-learning entering the semester, all measures of which improved as a result of their experiences in the course. In comparison with traditional courses, students felt service-learning courses have more societal benefits and should be increasingly offered. Participating in the service-learning project significantly increased students’ comfort working with people from different cultures or backgrounds and empowered them in different ways, including with decision-making. Ultimately, our data indicate that students enjoyed service-learning and received soft skill development through the course. We recommend service-learning be increasingly integrated into post-secondary curriculum.

Studies have found that educational credential attainment could reduce recidivism-- committing a criminal offense after being released from incarceration--by a significant amount (Davis et al., 2013; Lee College, 2019; Northwestern, 2020). Therefore, the philosophy department needed to create a program that would increase high-school-equivalency certification (GED, HiSet, etc.) attainment. The study of philosophy has shown high correlation with improved test scores and higher averages for writing and quantitative/ verbal reasoning (Topping & Trickery, 2007 and Education Testing Service, 2020.). A university held an event that focused on local community activism. One dialogue covered assistance programs for juveniles who are incarcerated or paroled. A local juvenile detention center leader approached a philosophy graduate student at the university, and plans for a philosophy-focused learning support pilot for juveniles began. As members of the philosophy department and in conjunction with the juvenile detention center, developed and led the pilot with a goal of (a) reducing recidivism and (b) increasing program completion and college interest.

Abstract Immunoglobulin G (IgG) antibodies contain a complex N-glycan embedded in the hydrophobic pocket between its heavy chain protomers. This glycan contributes to the structural organization of the Fc domain and determines its specificity for Fcγ receptors, thereby dictating distinct cellular responses. The variable construction of this glycan structure leads to highly-related, but non-equivalent glycoproteins known as glycoforms. We previously reported synthetic nanobodies that distinguish IgG glycoforms. Here, we present the structure of one such nanobody, X0, in complex with the Fc fragment of afucosylated IgG1. Upon binding, the elongated CDR3 loop of X0 undergoes a conformational shift to access the buried N-glycan and acts as a ‘glycan sensor’, forming hydrogen bonds with the afucosylated IgG N-glycan that would otherwise be sterically hindered by the presence of a core fucose residue. Based on this structure, we designed X0 fusion constructs that disrupt pathogenic afucosylated IgG1-FcγRIIIa interactions and rescue mice in a model of dengue virus infection.

As online learning continues to rise, students find educational value in consistency of instruction and technological support. With the significant increase in faculty designing courses, success lies with faculty education in instructional design. Additionally, students need support to navigate through the LMS and course elements to achieve success and increase their knowledge level. Determining the needs of nursing students and faculty is essential to the process of developing an effective course design for students leading to valuable content delivery and achievement of student outcomes. The purposes of this study were: (1) to measure attitudes toward online course design, (2) to compare online course delivery with instructor involvement, and (3) to determine changes to online courses. A significant finding is consistency within the LMS in course design and increased use of technology in content delivery.

Several candidate HIV subunit vaccines based on recombinant envelope (Env) glycoproteins have been advanced into human clinical trials. To facilitate biopharmaceutical production, it is necessary to produce these in CHO (Chinese Hamster Ovary) cells, the cellular substrate used for the manufacturing of most recombinant protein therapeutics. However, previous studies have shown that when recombinant Env proteins from clade B viruses, the major subtype represented in North America, Europe, and other parts of the world, are expressed in CHO cells, they are proteolyzed and lack important glycan-dependent epitopes present on virions. Previously, we identified C1s, a serine protease in the complement pathway, as the endogenous CHO protease responsible for the cleavage of clade B laboratory isolates of -recombinant gp120s (rgp120s) expressed in stable CHO-S cell lines. In this paper, we describe the development of two novel CHOK1 cell lines with the C1s gene inactivated by gene editing, that are suitable for the production of any protein susceptible to C1s proteolysis. One cell line, C1s-/- CHOK1 2.E7, contains a deletion in the C1s gene. The other cell line, C1s-/- MGAT1- CHOK1 1.A1, contains a deletion in both the C1s gene and the MGAT1 gene, which limits glycosylation to mannose-5 or earlier intermediates in the N-linked glycosylation pathway. In addition, we compare the substrate specificity of C1s with thrombin on the cleavage of both rgp120 and human Factor VIII, two recombinant proteins known to undergo unintended proteolysis (clipping) when expressed in CHO cells. Finally, we demonstrate the utility and practicality of the C1s-/- MGAT1- CHOK1 1.A1 cell line for the expression of clinical isolates of clade B Envs from rare individuals that possess broadly neutralizing antibodies and are able to control virus replication without anti-retroviral drugs (elite neutralizer/controller phenotypes). The Envs represent unique HIV vaccine immunogens suitable for further immunogenicity and efficacy studies.

Distinguishing biotic compounds from abiotic ones is critical to the search for life in the universe. Here, the authors demonstrate that the abiotic ethane has distinctively low 13C-13C abundances compared to biotic ethane.

IntroductionSoluble antigens complexed with immunoglobulin G (IgG) antibodies can induce robust adaptive immune responses in vitro and in animal models of disease. Factor VIII immune complexes (FVIII-ICs) have been detected in individuals with hemophilia A and severe von Willebrand disease following FVIII infusions. Yet, it is unclear if and how FVIII-ICs affect antibody development over time.MethodsIn this study, we analyzed internalization of FVIII complexed with epitope-mapped FVIII-specific IgG monoclonal antibodies (MAbs) by murine bone marrow-derived dendritic cells (BMDCs) in vitro and antibody development in hemophilia A (FVIII-/-) mice injected with FVIII-IC over time.ResultsFVIII complexed with 2-116 (A1 domain MAb), 2-113 (A3 domain MAb), and I55 (C2 domain MAb) significantly increased FVIII uptake by BMDC but only FVIII/2-116 enhanced antibody titers in FVIII-/- mice compared to FVIII alone. FVIII/4A4 (A2 domain MAb) showed similar FVIII uptake by BMDC to that of isolated FVIII yet significantly increased antibody titers when injected in FVIII-/- mice. Enhanced antibody responses observed with FVIII/2-116 and FVIII/4A4 complexes in vivo were abrogated in the absence of the FVIII carrier protein von Willebrand factor.ConclusionThese findings suggest that a subset of FVIII-IC modulates the humoral response to FVIII in an epitope-dependent manner, which may provide insight into the antibody response observed in some patients with hemophilia A.

Noble gases confirm billion-year groundwater residence times and external fluxes in deep crustal settings globally with implications for subsurface habitability and economic reservoir formation over planetary timescales both on Earth and beyond

The subsurface production, accumulation, and cycling of hydrogen (H2), and cogenetic elements such as sulfate (SO42-) and the noble gases (e.g., 4He, 40Ar) remains a critical area of research in the 21st century. Understanding how these elements generate, migrate, and accumulate is essential in terms of developing hydrogen as an alternative low-carbon energy source and as a basis for helium exploration which is urgently needed to meet global demand of this gas used in medical, industrial, and research fields. Beyond this, understanding the subsurface cycles of these compounds is key for investigating chemosynthetically-driven habitability models with relevance to the subsurface biosphere and the search for life beyond Earth. The challenge is that to evaluate each of these critical element cycles requires quantification and accurate estimates of production rates. The natural variability and intersectional nature of the critical parameters controlling production for different settings (local estimates), and for the planet as a whole (global estimates) are complex. To address this, we propose for the first time a Monte Carlo based approach which is capable of simultaneously incorporating both random and normally distributed ranges for all input parameters. This approach is capable of combining these through deterministic calculations to determine both the most probable production rates for these elements for any given system as well as defining upper and lowermost production rates as a function of probability and the most critical variables. This approach, which is applied to the Kidd Creek Observatory to demonstrate its efficacy, represents the next-generation of models which are needed to effectively incorporate the variability inherent to natural systems and to accurately model H2, 4He, 40Ar, SO42- production on Earth and beyond.

Recent measurements of methane (CH4) by the Mars Science Laboratory (MSL) now confront us with robust data that demand interpretation. Thus far, the MSL data have revealed a baseline level of CH4 (∼0.4 parts per billion by volume [ppbv]), with seasonal variations, as well as greatly enhanced spikes of CH4 with peak abundances of ∼7 ppbv. What do these CH4 revelations with drastically different abundances and temporal signatures represent in terms of interior geochemical processes, or is martian CH4 a biosignature? Discerning how CH4 generation occurs on Mars may shed light on the potential habitability of Mars. There is no evidence of life on the surface of Mars today, but microbes might reside beneath the surface. In this case, the carbon flux represented by CH4 would serve as a link between a putative subterranean biosphere on Mars and what we can measure above the surface. Alternatively, CH4 records modern geochemical activity. Here we ask the fundamental question: how active is Mars, geochemically and/or biologically? In this article, we examine geological, geochemical, and biogeochemical processes related to our overarching question. The martian atmosphere and surface are an overwhelmingly oxidizing environment, and life requires pairing of electron donors and electron acceptors, that is, redox gradients, as an essential source of energy. Therefore, a fundamental and critical question regarding the possibility of life on Mars is, "Where can we find redox gradients as energy sources for life on Mars?" Hence, regardless of the pathway that generates CH4 on Mars, the presence of CH4, a reduced species in an oxidant-rich environment, suggests the possibility of redox gradients supporting life and habitability on Mars. Recent missions such as ExoMars Trace Gas Orbiter may provide mapping of the global distribution of CH4. To discriminate between abiotic and biotic sources of CH4 on Mars, future studies should use a series of diagnostic geochemical analyses, preferably performed below the ground or at the ground/atmosphere interface, including measurements of CH4 isotopes, methane/ethane ratios, H2 gas concentration, and species such as acetic acid. Advances in the fields of Mars exploration and instrumentation will be driven, augmented, and supported by an improved understanding of atmospheric chemistry and dynamics, deep subsurface biogeochemistry, astrobiology, planetary geology, and geophysics. Future Mars exploration programs will have to expand the integration of complementary areas of expertise to generate synergistic and innovative ideas to realize breakthroughs in advancing our understanding of the potential of life and habitable conditions having existed on Mars. In this spirit, we conducted a set of interdisciplinary workshops. From this series has emerged a vision of technological, theoretical, and methodological innovations to explore the martian subsurface and to enhance spatial tracking of key volatiles, such as CH4.

As online learning continues to rise, students find educational value in consistency of instruction and technological support. With the significant increase in faculty designing courses, success lies with faculty education in instructional design. Additionally, students need support to navigate through the LMS and course elements to achieve success and increase their knowledge level. Determining the needs of nursing students and faculty is essential to the process of developing an effective course design for students leading to valuable content delivery and achievement of student outcomes. The purposes of this study were: (1) to measure attitudes toward online course design, (2) to compare online course delivery with instructor involvement, and (3) to determine changes to online courses. A significant finding is consistency within the LMS in course design and increased use of technology in content delivery.

Discussions surrounding incarcerated juveniles, education programs, and recidivism are difficult to find. In fact, gathering data is incredibly difficult in juvenile facilities because of high turnover rates and difficult of obtaining permission to conduct studies in juvenile facilities (Calderone, Bennett, Homan, Dedrick, & Chatfield, 2009; Shippen, Morton, Flynt, Houchins, & Smitherman, 2012). A study by David, Bozick, Steele, Saunders, and Miles found that participating in a GED or equivalent program reduced recidivism rates by 30% (2013). Assuming there is a relationship between juvenile recidivism rates and education programs, an important question remains. What types of education work best, and why? We believe a corequisite model paired with a philosophy course may be an important piece to this question.

Diffusion is a fundamental process in biological systems that governs the molecular collisions driving biochemical reactions and membrane and transport. Measurement of the diffusion coefficient and application of the Stokes-Einstein equation produces the hydrodynamic radius, which is a commonly used gauge of particle size. Additionally, measurement of the diffusion coefficient and the sedimentation coefficient, and application of the Svedberg equation, yields the molecular weight, which is particularly useful in the characterization of very large macromolecules. Dynamic light scattering (DLS) is the most common method to measure the diffusion coefficient of macromolecules. We describe a procedure to perform DLS measurements on monomeric bovine serum albumin (BSA) purified by size-exclusion chromatography using the Zetasizer Nano S particle size analyzer. We compare several analytical methods in existing software programs to estimate the diffusion coefficient of BSA (extrapolated to water at 20°C at infinite dilution, ) and describe a statistical method to obtain 95% confidence limits of the precision of the estimates. We compare estimates to literature values obtained by diffusiometry, sedimentation velocity analytical ultracentrifugation, and other DLS instruments. The method of cumulant analysis in the program SEDFIT (www.analyticalultracentrifugation.com) produced the most precise estimate, 6.06 ± 0.07 F (1 F = 10-7 cm2 s-1), which was within the range of estimates obtained by diffusiometry or sedimentation velocity. This protocol is useful for DLS method validation and quality control.

For the past fifty years of space exploration, mass spectrometry has provided unique chemical and physical insights on the characteristics of other planetary bodies in the Solar System. A variety of mass spectrometer types, including magnetic sector, quadrupole, time-of-flight, and ion trap, have and will continue to deepen our understanding of the formation and evolution of exploration targets like the surfaces and atmospheres of planets and their moons. An important impetus for the continuing exploration of Mars, Europa, Enceladus, Titan, and Venus involves assessing the habitability of solar system bodies and, ultimately, the search for life—a monumental effort that can be advanced by mass spectrometry. Modern flight-capable mass spectrometers, in combination with various sample processing, separation, and ionization techniques enable sensitive detection of chemical biosignatures. While our canonical knowledge of biosignatures is rooted in Terran-based examples, agnostic approaches in astrobiology can cast a wider net, to search for signs of life that may not be based on Terran-like biochemistry. Here, we delve into the search for extraterrestrial chemical and morphological biosignatures and examine several possible approaches to agnostic life detection using mass spectrometry. We discuss how future missions can help ensure that our search strategies are inclusive of unfamiliar life forms.

Abstract Global groundwater volumes in the upper 2 km of the Earth's continental crust—critical for water security—are well estimated. Beyond these depths, a vast body of largely saline and non‐potable groundwater exists down to at least 10 km—a volume that has not yet been quantified reliably at the global scale. Here, we estimate the amount of groundwater present in the upper 10 km of the Earth's continental crust by examining the distribution of sedimentary and crystalline rocks with depth and applying porosity‐depth relationships. We demonstrate that groundwater in the 2–10 km zone (what we call “deep groundwater”) has a volume comparable to that of groundwater in the upper 2 km of the Earth's crust. These new estimates make groundwater the largest continental reservoir of water, ahead of ice sheets, provide a basis to quantify geochemical cycles, and constrain the potential for large‐scale isolation of waste fluids.

Comparative studies on community phylogenetics and phylogeography of microorganisms living in extreme environments are rare. Terrestrial subsurface habitats are valuable for studying microbial biogeographical patterns due to their isolation and the restricted dispersal mechanisms. Since the taxonomic identity of a microorganism does not always correspond well with its functional role in a particular community, the use of taxonomic assignments or patterns may give limited inference on how microbial functions are affected by historical, geographical and environmental factors. With seven metagenomic libraries generated from fracture water samples collected from five South African mines, this study was carried out to (1) screen for ubiquitous functions or pathways of biogeochemical cycling of CH4, S, and N; (2) to characterize the biodiversity represented by the common functional genes; (3) to investigate the subsurface biogeography as revealed by this subset of genes; and (4) to explore the possibility of using metagenomic data for evolutionary study. The ubiquitous functional genes are NarV, NPD, PAPS reductase, NifH, NifD, NifK, NifE, and NifN genes. Although these eight common functional genes were taxonomically and phylogenetically diverse and distinct from each other, the dissimilarity between samples did not correlate strongly with geographical or environmental parameters or residence time of the water. Por genes homologous to those of Thermodesulfovibrio yellowstonii detected in all metagenomes were deep lineages of Nitrospirae, suggesting that subsurface habitats have preserved ancestral genetic signatures that inform the study of the origin and evolution of prokaryotes.

Factor VIII (fVIII) is a procoagulant protein that binds to activated factor IX (fIXa) on platelet surfaces to form the intrinsic tenase complex. Due to the high immunogenicity of fVIII, generation of antibody inhibitors is a common occurrence in patients during hemophilia A treatment and spontaneously occurs in acquired hemophilia A patients. Non-classical antibody inhibitors, which block fVIII activation by thrombin and formation of the tenase complex, are the most common anti-C2 domain pathogenic inhibitors in hemophilia A murine models and have been identified in patient plasmas. In this study, we report on the X-ray crystal structure of a B domain-deleted bioengineered fVIII bound to the non-classical antibody inhibitor, G99. While binding to G99 does not disrupt the overall domain architecture of fVIII, the C2 domain undergoes an ~8 Å translocation that is concomitant with breaking multiple domain-domain interactions. Analysis of normalized B-factor values revealed several solvent-exposed loops in the C1 and C2 domains which experience a decrease in thermal motion in the presence of inhibitory antibodies. These results enhance our understanding on the structural nature of binding non-classical inhibitors and provide a structural dynamics-based rationale for cooperativity between anti-C1 and anti-C2 domain inhibitors.

Home vegetable gardening has numerous documented benefits, including savings on the family food bill. How can a gardener calculate cost savings from a garden? Which vegetables cost more to grow in the garden and which cost less? This 10-page fact sheet written by Kevin Athearn, Hannah Wooten, Liz Felter, Catherine G. Campbell, Jessica M. Ryals, Matthew C. Lollar, Juanita Popenoe, Lorna Bravo, LuAnn Duncan, Christa Court, and Wendy Wilber and published by the UF/IFAS Food and Resource Economics Department helps home gardeners estimate the costs and cost savings from vegetable gardening

Deep subsurface environments are decoupled from Earth’s surface processes yet diverse, active, and abundant microbial communities thrive in these isolated environments. Microbes inhabiting the deep biosphere face unique challenges such as electron donor/acceptor limitations, pore space/fracture network limitations, and isolation from other microbes within the formation. Of the few systems that have been characterized, it is apparent that nutrient limitations likely facilitate diverse microbe-microbe interactions (i.e., syntrophic, symbiotic, or parasitic) and that these interactions drive biogeochemical cycling of major elements. Here we describe microbial communities living in low temperature, chemically reduced brines at the Soudan Underground Mine State Park, United States. The Soudan Iron mine intersects a massive hematite formation at the southern extent of the Canadian Shield. Fractured rock aquifer brines continuously flow from exploratory boreholes drilled circa 1960 and are enriched in deuterium compared to the global meteoric values, indicating brines have had little contact with surface derived waters, and continually degas low molecular weight hydrocarbons C1-C4. Microbial enrichments suggest that once brines exit the boreholes, oxidation of the hydrocarbons occur. Amplicon sequencing show these borehole communities are low in diversity and dominated by Firmicute and Proteobacteria phyla. From the metagenome assemblies, we recovered approximately thirty genomes with estimated completion over 50%. Analysis of genome taxonomy generally followed the amplicon data, and highlights that several of the genomes represent novel families and genera. Metabolic reconstruction shows two carbon-fixation pathways were dominant, the Wood-Ljungdahl (acetogenesis) and Calvin-Benson-Bassham (via RuBisCo), indicating that inorganic carbon likely enters into the microbial foodweb with differing carbon fractionation potentials. Interestingly, methanogenesis is likely driven by Methanolobus and suggests cycling of methylated compounds and not H2/CO2 or acetate. Furthermore, the abundance of sulfate in brines suggests cryptic sulfur cycling may occur, as we detect possible sulfate reducing and thiosulfate oxidizing microorganisms. Finally, a majority of the microorganisms identified contain genes that would allow them to participate in several element cycles, highlighting that in these deep isolated systems metabolic flexibility may be an important life history trait.