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TO DETERMINE THE FEASIBILITY OF INTEGRATING BIOLOGICAL PRINCIPLES WITH INSTRUCTION IN VOCATIONAL AGRICULTURE IN OHIO HIGH SCHOOLS, 15 PILOT SCHOOLS AND 8 CONTROL SCHOOLS WERE STUDIED. PRETESTS ADMINISTERED TO STUDENTS IN OCTOBER 1963 WERE AN AGRICULTURAL ACHIEVEMENT TEST, A BIOLOGICAL PRINCIPLES TEST, THE CALIFORNIA SHORT-FORM TEST OF MENTAL MATURITY, AND THE EDUCATIONAL INTEREST INVENTORY. POSTTESTING IN MAY 1964 INCLUDED ALL EXCEPT THE CALIFORNIA SHORT-FORM. A QUESTIONNAIRE ADMINISTERED TO AGRICULTURE TEACHERS IN THE PILOT SCHOOLS REVEALED EIGHT ADVANTAGES, INCLUDING--(1) THE PROGRAM WAS MORE CHALLENGING AND INTERESTING TO STUDENTS, AND (2) IT CAUSED THE TEACHER TO PREPARE BETTER. ONE OF THE FOUR DISADVANTAGES WAS THAT MORE TIME WAS NECESSARY TO TEACH BIOLOGICAL PRINCIPLES, AND OTHER UNITS HAD TO BE DROPPED. INTERPRETATION OF THE TEST DATA WAS THE BASIS FOR EIGHT CONCLUSIONS, INCLUDING--(1) THE NUMBER OF PRINCIPLES TAUGHT WAS THE MOST SIGNIFICANT FACTOR INFLUENCING ACHIEVEMENT IN BIOLOGY, (2) DIFFERENCE IN STUDENT ACHIEVEMENT BETWEEN 1ST- AND 2ND-YEAR PILOT SCHOOLS WAS NOT CONSISTENT ON ALL TESTS, AND (3) FRESHMAN AND SOPHOMORE STUDENTS SHOWED GREATER ACHIEVEMENT THAN JUNIOR AND SENIOR STUDENTS WHEN THE BIOLOGICAL PRINCIPLES APPROACH WAS USED. (EM)

Describes procedures to develop instructional objectives for high school biology. Two kinds of objectives are identified as pre-objectives and performance objectives. Models to classify these in branches of biology and to ensure quality control are provided. (PS)

Aims The study of microbial inactivation rates in aquifer systems has most often been determined in aerobic and oxidized systems. This study examined the inactivation (i.e. loss of infectivity) of MS2, poliovirus type 1 (PV1) and Cryptosporidium parvum in an anaerobic and reduced groundwater system that has been identified as storage zones for aquifer storage and recovery (ASR) facilities. Methods and Results Anaerobic and reduced (ORP < −250 mV) groundwater from an artesian well was diverted to an above-ground, flow-through mesocosm that contained diffusion chambers filled with MS2, PV1 or Cryptosporidium parvum. The respective infectivity assays were performed on microorganisms recovered from the diffusion chambers during 30- to 58-day experiments. The net reduction in infectivity was 5.73 log10 over 30 days for MS2, 5.00 log10 over 58 days for PV1 and 4.07 log10 over 37 days for C. parvum. The best fit inactivation model for PV1 was the log-linear model and the Weibull model for MS2 and C. parvum, with respective inactivation rates (95% confidence interval) of 0.19 (0.17–0.21) log10 day−1, 0.31 (0.19–0.89) log10 day−1 and 0.20 (0.14–0.37) log10 day−1. Conclusions The groundwater geochemical conditions in this aquifer enhanced the inactivation of MS2, PV1, and C. parvum at rates approximately 2.0–5.3-fold, 1.2–17.0-fold, and 4.5–5.6-fold greater, respectively, than those from published studies that used diffusion chambers in aerobic-to-anoxic groundwater systems, with positive redox potentials. Significance and Impact of the Study Geochemical conditions like those in the aquifer zone in this study can naturally and significantly reduce concentrations of microbial indicators and pathogens of human health concern in injected surface water. Appropriate storage times for injected surface water could complement above-ground engineered processes for microorganism removal and inactivation (e.g. filtration, disinfection) by naturally increasing overall microorganism log-inactivation rates of ASR facilities.

The Mesoproterozoic Rossport Formation of Ontario, Canada is generally interpreted as having been deposited in an intracratonic basin, most probably a rift‐related lake. While the Rossport, overall, is dominated by sandstone and shale, the Middlebrun Bay Member, in the middle of the formation, is a carbonate unit. The Middlebrun Bay Member, in exposures on the Channel Islands and along the north Shore of Lake Superior, consists most commonly of cherty, dolomitic microbial laminites and low‐relief columnar to conical forms. In contrast to typical outcrops, the Middlebrun Bay Member on Copper Island expresses as a massive, coarsely crystalline limestone unit, devoid of stromatolites or microbial laminae. Several features suggest dissolution and replacement of a primary, soluble phase such as an evaporite mineral. The top of the unit is marked by evidence of dissolution and collapse, including large sandstone clasts let down from the overlying bed. At petrographic scale, the Copper Island Carbonate comprises millimetre‐scale anhedral spar with abundant stylolites and concentration of insoluble material at grain boundaries, indicating recrystallization from a previous phase. Geochemical data from Copper Island and from correlative stromatolitic carbonate on Channel Island and mainland Ontario are consistent with hypersaline conditions, with elevated concentrations of carbonate‐associated sulphate, V and Ba. Based on these data, the massive carbonate exposed on Copper Island is interpreted as a calcitized evaporite, probably deposited originally as gypsum, and replaced by calcite during diagenesis. These data support previous work suggesting that the Middlebrun Bay interval was deposited during a period of increased aridity and low clastic influx, and further suggest that this restricted, hypersaline lake precipitated both carbonate and gypsum, comparable to modern arid playa lakes.

There are many different types of berry-producing plants that grow throughout North America. These various species of plants produce berries, known by their common names, such as blueberry, lingonberry, cranberry, crowberry, blackberry, bunchberry, bearberry, and strawberry. The berry fruits are important food sources and can be eaten fresh or preserved. The leaves of several species can also be enjoyed in teas. Contemporary medicine has demonstrated many potential health benefits of berries due to their antioxidant and anti-inflammatory activity. Indigenous Peoples have not only utilized berries as food, likely over many centuries, but have also passed down knowledge on the medicinal uses of various parts of these plants including the fruits, leaves and roots. This short review provides a synopsis of some of the major species of berry-producing plants that are prevalent in Canada, particularly eastern Canada. Current knowledge and traditional uses of important species of berry-producing plants are discussed.

This review describes an Indigenous-led project run by Q’eqchi’ Maya Healers of Belize meant to strengthen and improve traditional botanical healing. The goals of this project were to conserve medicinal plant knowledge by way of ethnobotanical studies, and to conserve the plants themselves by creating a community ethnobotanical garden. A total of 169 medicinal species were collected in the ethnobotanical survey, which provided unique knowledge on many rainforest species of the wet lowland forest of southern Belize, not found in neighbouring Indigenous cultures. Consensus on plant uses by the Healers was high, indicating a well-conserved, codified oral history. After horticultural experimentation by the Healers, the Indigenous botanical garden provided a habitat for and conservation of 102 medicinal species including many epiphytes that were rescued from forested areas. Ethnopharmacological studies by the university partners showed a pharmacological basis for, and active principles of, plants used for epilepsy and anxiety, for inflammatory conditions such as arthritis, for dermatological mycoses, and for type 2 diabetes complications. Overall, the project has provided a model for Indigenous empowerment and First Nation’s science, as well as establishing traditional medicine as an important, unified healing practice that can safely and effectively provide primary health care in its cultural context.

Background and AimsSingle-cell transcriptomics offer unprecedented resolution of tissue function at the cellular level, yet studies analyzing healthy adult human small intestine and colon are sparse. Here, we present single-cell transcriptomics covering the duodenum, jejunum, ileum, and ascending, transverse, and descending colon from 3 humans.Methods12,590 single epithelial cells from three independently processed organ donors were evaluated for organ-specific lineage biomarkers, differentially regulated genes, receptors, and drug targets. Analyses focused on intrinsic cell properties and capacity for response to extrinsic signals along the gut axis across different humans.ResultCells were assigned to 25 epithelial lineage clusters. Human intestinal stem cells (ISCs) are not specifically marked by many murine ISC markers. Lysozyme expression is not unique to human Paneth cells (PCs), and PCs lack expression of expected niche-factors. BEST4+ cells express NPY and show maturational differences between SI and colon. Tuft cells possess a broad ability to interact with the innate and adaptive immune systems through previously unreported receptors. Some classes of mucins, hormones, cell-junction, and nutrient absorption genes show unappreciated regional expression differences across lineages. Differential expression of receptors and drug targets across lineages reveals biological variation and potential for variegated responses.ConclusionsOur study identifies novel lineage marker genes; covers regional differences; shows important differences between mouse and human gut epithelium; and reveals insight into how the epithelium responds to the environment and drugs. This comprehensive cell atlas of the healthy adult human intestinal epithelium resolves likely functional differences across anatomical regions along the gastrointestinal tract and advances our understanding of human intestinal physiology.

Eastern equine encephalitis virus (EEEV), western equine encephalitis virus (WEEV) and Venezuelan equine encephalitis virus (VEEV) can cause fatal encephalitis in humans and equids. Some MAbs to the E1 glycoprotein are known to be cross-reactive, weakly neutralizing in vitro but can protect from disease in animal models. We investigated the mechanism of neutralization of VEEV infection by the broadly cross-reactive E1-specific MAb 1A4B-6. 1A4B-6 protected 3-week-old Swiss Webster mice prophylactically from lethal VEEV challenge. Likewise, 1A4B-6 inhibited virus growth in vitro at a pre-attachment step after virions were incubated at 37 °C and inhibited virus-mediated cell fusion. Amino acid residue N100 in the fusion loop of E1 protein was identified as critical for binding. The potential to elicit broadly cross-reactive MAbs with limited virus neutralizing activity in vitro but that can inhibit virus entry and protect animals from infection merits further exploration for vaccine and therapeutic developmental research.

The European species of the potter wasp genus Eumenes Latreille, 1802 (Vespidae, Eumeninae) are illustrated and a new illustrated key to the 13 recognised species is presented. Eumenes mediterraneus aemilianus Guiglia, 1951 is synonymised with E. papillarius (Christ, 1791) (syn. nov.), E. obscurus André, 1884 and E. andrei Dalla Torre, 1894 with E. pedunculatus (Panzer, 1799) (syn. nov.) and E. crimensis Blüthgen, 1938 with E. sareptanus André, 1884 (syn. nov.).

Lineage plasticity is implicated in treatment resistance in multiple cancers. In lung adenocarcinomas (LUAD) amenable to targeted therapy, transformation to small cell lung cancer (SCLC) is a recognized resistance mechanism. Defining molecular mechanisms of neuroendocrine (NE) transformation in lung cancer has been limited by a paucity of pre/posttransformation clinical samples. Detailed genomic, epigenomic, transcriptomic, and protein characterization of combined LUAD/SCLC tumors, as well as pre/posttransformation samples, supports that NE transformation is primarily driven by transcriptional reprogramming rather than mutational events. We identify genomic contexts in which NE transformation is favored, including frequent loss of the 3p chromosome arm. We observed enhanced expression of genes involved in the PRC2 complex and PI3K/AKT and NOTCH pathways. Pharmacologic inhibition of the PI3K/AKT pathway delayed tumor growth and NE transformation in an EGFR-mutant patient-derived xenograft model. Our findings define a novel landscape of potential drivers and therapeutic vulnerabilities of NE transformation in lung cancer. Significance: The difficulty in collection of transformation samples has precluded the performance of molecular analyses, and thus little is known about the lineage plasticity mechanisms leading to LUAD-to-SCLC transformation. Here, we describe biological pathways dysregulated upon transformation and identify potential predictors and potential therapeutic vulnerabilities of NE transformation in the lung. See related commentary by Meador and Lovly, p. 2962. This article is highlighted in the In This Issue feature, p. 2945

Chondrosarcomas are inherently resistant to chemotherapy and radiotherapy, pointing to an unmet need for new treatment options. Immune checkpoint inhibitors, which have shown remarkable promise in multiple solid cancer types, have limited efficacy in chondrosarcomas. Mutations in IDH1/2 genes, which result in progressive increases in DNA and histone methylation, are observed in 50% of conventional chondrosarcomas, suggesting that epigenetic dysregulation represents a potential barrier for tumor progression and target for therapeutic intervention. Here, we demonstrated that combined treatment of FDA-approved inhibitors of DNA methyltransferases (DNMTs) 5-aza-2′-deoxycytidine (5-aza), and histone deacetylases (HDACs) suberanilohydroxamic acid (SAHA) impaired the proliferation of chondrosarcoma cell lines in vitro and in xenograft studies. Transcriptomic analysis reveals that chondrosarcoma cells treated with 5-aza and SAHA markedly elevated the expression of IFN-stimulated genes including PD-L1, indicating that these epigenetic drugs induced a potent innate immune response. We demonstrated that 5-aza and SAHA resulted in both genomic and epigenomic instability, as shown by elevated DNA damage response and derepression of retrotransposons, respectively, which in turn activated pattern recognition receptors (PRRs) and the downstream IFN signaling pathways. Importantly, the cytotoxic effects of 5-aza and SAHA can be rescued by depletion of PRRs such as cGAS and MAVS, and potentiated by depletion of the RNA-editing enzyme ADAR1. Together, our results demonstrate preclinical activity of combined DNMT and HDAC inhibition against chondrosarcomas and suggest that targeted epigenetic therapies could represent a new therapeutic approach in the treatment of chondrosarcomas, and this is being tested in an ongoing clinical trial (NCT04340843).

Tour guides play a central role in the implementation of sustainable tourism practices through maximizing tourists’ appreciation and enjoyment of the destination and minimizing their negative impac...

In the skin-healing field, porcine models are regarded as a useful analogue for human skin due to their numerous anatomical and physiological similarities. Despite the widespread use of porcine models in skin healing studies, the initial origin, recruitment and transition of fibroblasts to matrix-secreting contractile myofibroblasts are not well defined for this model. In this review, we discuss the merit of the pig as an animal for studying myofibroblast origin, as well as the challenges associated with assessing their contributions to skin healing. Although a variety of wound types (incisional, partial thickness, full thickness, burns) have been investigated in pigs in attempts to mimic diverse injuries in humans, direct comparison of human healing profiles with regards to myofibroblasts shows evident differences. Following injury in porcine models, which often employ juvenile animals, myofibroblasts are described in the developing granulation tissue at 4 days, peaking at Days 7-14, and persisting at 60 days post-wounding, although variations are evident depending on the specific pig breed. In human wounds, the presence of myofibroblasts is variable and does not correlate with the age of the wound or clinical contraction. Our comparison of porcine myofibroblast-mediated healing processes with those in humans suggests that further validation of the pig model is essential. Moreover, we identify several limitations evident in experimental design that need to be better controlled, and standardisation of methodologies would be beneficial for the comparison and interpretation of results. In particular, we discuss anatomical location of the wounds, their size and depth, as well as the healing microenvironment (wet vs. moist vs. dry) in pigs and how this could influence myofibroblast recruitment. In summary, although a widespread model used in the skin healing field, further research is required to validate pigs as a useful analogue for human healing with regards to myofibroblasts.

As the human footprint upon the landscape expands, wildlife seeking to avoid human contact are losing the option of altering their spatial distribution and instead are shifting their daily activity patterns to be active at different times than humans. In this study, we used game cameras to evaluate how human development and activity were related to the daily activity patterns of the nine‐banded armadillo (Dasypus novemcinctus) along an urban to rural gradient in Arkansas, USA during the winter of 2020–2021. We found that armadillos had substantial behavioral plasticity in regard to the timing of their activity patterns; >95% of armadillo activity was nocturnal at six of the study sites, whereas between 30% and 60% of activity occurred during the day at three other sites. The likelihood of diurnal armadillo activity was best explained by the distance to downtown Fayetteville (the nearest population center) and estimated ambient sound level (both indices of human activity) with armadillos being most active during the day at quiet sites far from Fayetteville. Furthermore, armadillo activity occurred later during the night period (minutes after sunset) at sites near downtown and with higher anthropogenic sound. Anecdotal evidence suggests that the observed activity shift may be in response to not only human activity but also the presence of domestic dogs. Our results provide further evidence that human activity has subtle nonlethal impacts on even common, widespread wildlife species. Because armadillos have low body temperatures and basal metabolism, being active during cold winter nights likely has measurable fitness costs. Nature reserves near human population centers may not serve as safe harbors for wildlife as we intend, and managers could benefit from considering these nonlethal responses in how they manage recreation and visitation in these natural areas., The nine‐banded armadillo (Daspyus novemcinctus) is a widespread mammal capable of being active during the day or night. Here, we present evidence that armadillos living in proximity to humans shift their activity to be nocturnal despite likely thermal fitness costs to avoid human contact.

BACKGROUND: Recruitment and activation of brown adipose tissue (BAT) results in increased energy expenditure (EE) via thermogenesis and represents an intriguing therapeutic approach to combat obesity and treat associated diseases. Thermogenesis requires an increased and efficient supply of energy substrates and oxygen to the BAT. The hemoprotein myoglobin (MB) is primarily expressed in heart and skeletal muscle fibres, where it facilitates oxygen storage and flux to the mitochondria during exercise. In the last years, further contributions of MB have been assigned to the scavenging of reactive oxygen species (ROS), the regulation of cellular nitric oxide (NO) levels and also lipid binding. There is a substantial expression of MB in BAT, which is induced during brown adipocyte differentiation and BAT activation. This suggests MB as a previously unrecognized player in BAT contributing to thermogenesis. METHODS AND RESULTS: This study analyzed the consequences of MB expression in BAT on mitochondrial function and thermogenesis in vitro and in vivo. Using MB overexpressing, knockdown or knockout adipocytes, we show that expression levels of MB control brown adipocyte mitochondrial respiratory capacity and acute response to adrenergic stimulation, signalling and lipolysis. Overexpression in white adipocytes also increases their metabolic activity. Mutation of lipid interacting residues in MB abolished these beneficial effects of MB. In vivo, whole-body MB knockout resulted in impaired thermoregulation and cold- as well as drug-induced BAT activation in mice. In humans, MB is differentially expressed in subcutaneous (SC) and visceral (VIS) adipose tissue (AT) depots, differentially regulated by the state of obesity and higher expressed in AT samples that exhibit higher thermogenic potential. CONCLUSIONS: These data demonstrate for the first time a functional relevance of MBs lipid binding properties and establish MB as an important regulatory element of thermogenic capacity in brown and likely beige adipocytes.

Funder: Fundamental Research Funds for the Central Universities (Grant No.16CX06050A)., Funder: Medical Research Council (MRC) grant (MR/T004347/1), Funder: Alzheimer’s Research UK in partnership with Hidden Hearing (ARUK-PPG2016A-2); National Institute of Health Research (NIHR) Biomedical Research Centre (BRC), Funder: National Natural Science Foundation of P.R. China (Grant No.62072468) and the National Natural Science Foundation of Shandong Province (Grant No. ZR2018MF017)., Funder: Alzheimer’s Research UK in partnership with Hidden Hearing (ARUK-PPG2016A-2); National Institute of Health Research (NIHR) Biomedical Research Centre (BRC); Medical Research Council (MRC) grant (MR/T004347/1); Guangci Professorship Program of Ruijin Hospital (Shanghai Jiao Tong University), Funder: Alzheimer’s Research UK in partnership with Hidden Hearing (ARUK-PPG2016A-2); National Institute of Health Research (NIHR) Biomedical Research Centre (BRC); Wellcome Trust Fellowship (WT088441MA), Brain’s modular connectivity gives this organ resilience and adaptability. The ageing process alters the organised modularity of the brain and these changes are further accentuated by neurodegeneration, leading to disorganisation. To understand this further, we analysed modular variability—heterogeneity of modules—and modular dissociation—detachment from segregated connectivity—in two ageing cohorts and a mixed cohort of neurodegenerative diseases. Our results revealed that the brain follows a universal pattern of high modular variability in metacognitive brain regions: the association cortices. The brain in ageing moves towards a segregated modular structure despite presenting with increased modular heterogeneity—modules in older adults are not only segregated, but their shape and size are more variable than in young adults. In the presence of neurodegeneration, the brain maintains its segregated connectivity globally but not locally, and this is particularly visible in dementia with Lewy bodies and Parkinson’s disease dementia; overall, the modular brain shows patterns of differentiated pathology.

To develop an effective genome editing tool for blueberry breeding, CRISPR-Cas9 and CRISPR-Cas12a were evaluated for their editing efficiencies of a marker gene, beta-glucuronidase (gusA), which was previously introduced into two blueberry cultivars each a single-copy transgene. Four expression vectors were built, with CRISPR-Cas9 and CRISPR-Cas12a each driven by a 35S promoter or AtUbi promoter. Each vector contained two editing sites in the gusA. These four vectors were respectively transformed into the leaf explants of transgenic gusA blueberry and the resulting transgenic calli were induced under hygromycin selection. GUS staining showed that some small proportions of the hygromycin-resistant calli had non-GUS stained sectors, suggesting some possible occurrences of gusA editing. We sequenced GUS amplicons spanning the two editing sites in three blueberry tissues and found about 5.5% amplicons having editing features from the calli transformed with the 35S-Cas9 vector. Further, we conducted a second round of shoot regeneration from leaf explants derived from the initial Cas9- and Cas12a-containing calli (T0) and analyzed amplicons of the target editing region. Of the newly induced shoots, 15.5% for the 35S-Cas9 and 5.3% for the AtUbi-Cas9 showed non-GUS staining, whereas all of the shoots containing the Cas12a vectors showed blue staining. Sanger sequencing confirmed the editing-induced mutations in two representative non-GUS staining lines. Clearly, the second round of regeneration had enriched editing events and enhanced the production of edited shoots. The results and protocol described will be helpful to facilitating high-precision breeding of blueberries using CRISPR Cas technologies. A second round of regeneration enriched editing events and enhanced the production of edited blueberry shoots. The new protocol described facilitates high-precision breeding of blueberries using CRISPR Cas technologies.

Platystethynium (Platypatasson) earlyi Huber, sp. n. (Hymenoptera: Mymaridae), is described from both sexes reared from an egg of Rhaphidophoridae (Orthoptera) found in an old mining tunnel in New Zealand. The male is micropterous and has lateral ocelli but no median ocellus, a unique feature in Mymaridae found so far only in males of Platystethynium Ogloblin species. The remaining described Eastern Hemisphere species of Platystethynium are discussed and Platystethynium glabrum Jin & Li, syn. n., is placed in synonymy under P. onomarchicidum Ogloblin. A key to females, and males where known, of the described Eastern Hemisphere species of Platystethynium is given.

This editorial introduces the Australian Journal of Botany special issue ‘Vegetation science for decision-making’. Vegetation science and classification are crucial to understanding Australian landscapes. From the mulga shrublands of the arid interior to the monsoon rain forests of northern Australia, we have culturally and scientifically built upon the delineation of vegetation into recognisable and repeatable patterns. As remote sensing and database capacities increase, this improved capability to measure vegetation and share data also prompts collaboration and synthesis of complex, specialised datasets. Although the task faces significant challenges, the growing body of literature demonstrates a strong discipline. In Australia, purpose-driven products describe vegetation at broad scales (e.g. the National Vegetation Information System, the Terrestrial Ecosystem Research Network). At fine scales however (i.e. that of the vegetation community), no uniform framework or agreed protocols exist. Climate and landform dictate vegetation patterns at broad scales, but microtopography, microclimate and biotic processes act as filters at finer scales. This is the scale where climate-change impacts are most likely to be detected and effected; this is the scale at which a deeper understanding of evolutionary ecology will be achieved, and it is the scale at which species need to be protected. A common language and system for understanding Australian communities and impetus for collecting data at this scale is needed. In the face of ongoing climate and development pressures and an increasingly complex set of tools to manage these threats (e.g. offset policies, cumulative impact assessments), a nationally collaborative approach is needed. It is our hope that this special issue will help to achieve this.

Specialized diterpenoid metabolites are important mediators of plant-environment interactions in monocot crops. To understand metabolite functions in plant environmental adaptation that ultimately can enable crop improvement strategies, a deeper knowledge of the underlying species-specific biosynthetic pathways is required. Here, we report the genomics-enabled discovery of five cytochrome P450 monooxygenases (CYP71Z25-CYP71Z29) that form previously unknown furanoditerpenoids in the monocot bioenergy crop Panicum virgatum (switchgrass). Combinatorial pathway reconstruction showed that CYP71Z25-CYP71Z29 catalyze furan ring addition directly to primary diterpene alcohol intermediates derived from distinct class II diterpene synthase products. Transcriptional co-expression patterns and the presence of select diterpenoids in switchgrass roots support the occurrence of P450-derived furanoditerpenoids in planta. Integrating molecular dynamics, structural analysis and targeted mutagenesis identified active site determinants that contribute to the distinct catalytic specificities underlying the broad substrate promiscuity of CYP71Z25-CYP71Z29 for native and non-native diterpenoids.

A respiratory disease epizootic at the National Bison Range (NBR) in Montana in 2016–2017 caused an 85% decline in the bighorn sheep population, documented by observations of its unmarked but individually identifiable members, the subjects of an ongoing long‐term study. The index case was likely one of a small group of young bighorn sheep on a short‐term exploratory foray in early summer of 2016. Disease subsequently spread through the population, with peak mortality in September and October and continuing signs of respiratory disease and sporadic mortality of all age classes through early July 2017. Body condition scores and clinical signs suggested that the disease affected ewe groups before rams, although by the end of the epizootic, ram mortality (90% of 71) exceeded ewe mortality (79% of 84). Microbiological sampling 10 years to 3 months prior to the epizootic had documented no evidence of infection or exposure to Mycoplasma ovipneumoniae at NBR, but during the epizootic, a single genetic strain of M. ovipneumoniae was detected in affected animals. Retrospective screening of domestic sheep flocks near the NBR identified the same genetic strain in one flock, presumptively the source of the epizootic infection. Evidence of fatal lamb pneumonia was observed during the first two lambing seasons following the epizootic but was absent during the third season following the death of the last identified M. ovipneumoniae carrier ewe. Monitoring of life‐history traits prior to the epizootic provided no evidence that environmentally and/or demographically induced nutritional or other stress contributed to the epizootic. Furthermore, the epizootic occurred despite proactive management actions undertaken to reduce risk of disease and increase resilience in this population. This closely observed bighorn sheep epizootic uniquely illustrates the natural history of the disease including the (presumptive) source of spillover, course, severity, and eventual pathogen clearance., This case report provides unprecedentedly detailed animal observations before, during, and after a severe bighorn sheep pneumonia outbreak and includes molecular detection of the causative primary pathogen, Mycoplasma ovipneumoniae, in its putative source, a nearby domestic sheep operation. As a result, this case report serves to advance our understanding of critical aspects of this important disease.

Myzus persicae has evolved resistance to various insecticides in Greece. Here we examine the effectiveness of the insecticide flupyradifurone against aphid clones collected from tobacco and peach in Greece during 2017-2020. Furthermore, we monitored the frequency of the neonicotinoid resistance mutation R81T in the sampled clones, and the association between the responses to flupyradifurone and acetamiprid.Of 43 clones tested with flupyradifurone, 6.977%, 60.465% and 32.558% showed low (10-14), moderate (19-89) and high (104-1914) resistance factor (RF) values, respectively. Resistance was higher in clones from peach than from tobacco with 42.308% and 17.647% of clones (respectively) failing into the high RF category (median RF values 67.5 and 36.4 for clones from peach and tobacco, respectively). Acetamiprid resistance was detected in clones collected in 2019-2020, in line with our previous study in Greece. The analysis of the whole dataset (54 clones collected during 2017-2020) revealed that all tobacco clones had RF 7.5, whereas 55.263%, 18.421% and 26.316% of the peach clones exhibited low (12), moderate (20-48) and high (100-145) RF values, respectively. A significant but moderate association between flupyradifurone and acetamiprid responses was detected (r = 0.513, P 0.001). The R81T mutation was detected in aphids from peach (5.6% and 32.6% as homozygotes and heterozygotes, respectively) and in one aphid specimen (heterozygote) from tobacco. R81T was partially associated with the resistance to both insecticides, but many highly resistant clones did not possess the mutation, indicating the possible operation of one or more alternative underlying resistance mechanisms.The use of flupyradifurone and acetamiprid in IPM/IRM should be based on further ongoing susceptibility monitoring. © 2021 Society of Chemical Industry.

The global COVID-19 pandemic has created an urgent demand for large numbers of inexpensive, accurate, rapid, point-of-care diagnostic tests. Analyte-based assays are suitably rapid and inexpensive and can be rapidly mass-produced, but for sufficiently accurate performance, they require highly optimized antibodies and assay conditions. We used an automated liquid handling system, customized to handle arrays of lateral flow (immuno)assays (LFAs) in a high-throughput screen, to identify anti-nucleocapsid antibodies that will perform optimally in an LFA. We tested 1021 anti-nucleocapsid antibody pairs as LFA capture and detection reagents with the goal of highlighting pairs that have the greatest affinity for the nucleocapsid protein of SARS-CoV-2 within the LFA format. In contrast to traditional antibody screening methods (e.g., ELISA, bio-layer interferometry), the method described here integrates real-time reaction kinetics with transport in, and immobilization directly onto, nitrocellulose. We have identified several candidate antibody pairs that are suitable for further development of an LFA for SARS-CoV-2.

Separate identification keys for females, and for males where known, of the 40 genera of Mymaridae (Hymenoptera: Chalcidoidea) in the Afrotropical region are given. The subgenera of four genera are also included in the key to females. The genera are illustrated with over 300 photographs. The 122 named, valid species reported from the region are catalogued. Reliable host records are reported for 6 genera and 11 species from rearings undertaken in the region. An appendix lists the 27 species from northern Africa and offshore islands (Canary Islands, Madeira) but not recorded in the Afrotropical region as defined in this publication. Camptoptera (Zemicamptoptera) Ogloblin & Annecke, syn. n. is synonymized with Camptoptera (Camptoptera) Foerster. Eofoersteria Mathot, syn. n. is synonymized with Camptoptera Foerster and is treated as subgenus C. (Eofoersteria) Mathot, stat. n.; its type species Eofoersteria camptopteroides Mathot is transferred to Camptoptera as C. (Eofoersteria) camptopteroides (Mathot), comb. n. Polynema megacephala (Risbec) is transferred to Lymaenon as L. megacephala (Risbec), comb. n. Limacis opuntiae Risbec is transferred to Encarsia Foerster (Hymenoptera: Aphelinidae) as E. opuntiae (Risbec), comb. n. A few corrections to Huber et al. (2020) are given.

Heat treatment is one of the most widely used processing technologies in the dairy industry. Its primary purpose is to destroy microorganisms, both pathogenic and spoilage, to ensure the product is safe and has a reasonable shelf life. In this study microwave volumetric heating (MVH) was compared with a conventional tubular heat exchanger (THE), in terms of the effects of each at a range of temperatures (75°C, 85°C, 95°C, 105°C, 115°C, and 125°C) on indigenous microflora viability and the germination of inoculated Bacillus licheniformis endospores in reconstituted skim milk. To assess the heat treatment-related effects on microbial viability, classical agar-based tests were applied to obtain the counts of 4 various microbiological groups including total bacterial, thermophilic bacterial, mesophilic aerobic bacterial endospore, and thermophilic aerobic bacterial endospore counts, and additional novel insights into cell permeability and spore germination profiles post-heat treatment were obtained using real-time flow cytometry (FC) methods. No significant differences in the plate counts of the indigenous microorganisms tested, the plate counts of the inoculated B. licheniformis, or the relative percentage of germinating endospores were observed between MVH- and THE-treated samples, at equal temperatures in the range specified above, indicating that both methods inactivated inoculated endospores to a similar degree (up to 70% as measured by FC and 5 log reduction as measured by plate counting for some treatments of inoculated endospores). Furthermore, increased cell permeability of indigenous microflora was observed by FC after MVH compared with THE treatment of uninoculated skim milk, which was reflected in lower total bacterial count at a treatment temperature of 105°C. This work demonstrates the utility of FC as a rapid method for assessing cell viability and spore inactivation for postthermal processing in dairy products and overall provides evidence that MVH is at least as effective at eliminating native microflora and inoculated B. licheniformis endospores as THE.

Many previous vegetation description and mapping programs have highlighted the need for a hierarchical classification that easily translates from regional through to international scales. Often programs have not selected appropriate levels of vegetation classification and mapping for management purposes. Here, we provide an alternative broad vegetation classification at the macrogroup level in the international vegetation classification for eastern New South Wales (NSW) that could be applied in regional and local environmental planning and management. Existing aerial photographic interpretation (API) floristic and canopy maps available for eastern NSW were collated and reclassified and incorporated into a unifying dataset of macrogroups, which also included types dominated by exotic vegetation such as exotic plantations. There we identified, in total, 71 macrogroups across eastern New South Wales (NSW) of a total of 85 identified for NSW mapped at 1:25 000 scale, or finer, in the following three sections: north-east, Sydney basin and central west, and southern coast and Snowy Mountains. These were then amalgamated into a single remnant vegetation map covering 23 525 280 ha. This alternative broad vegetation hierarchy provides a reliable and overt vegetation classification and mapping dataset that can easily incorporate future revisions as new mapping data become available.

Glyphosate is the active ingredient in Roundup® brand nonselective herbicides, and residue testing for food has been conducted as part of the normal regulatory processes. Additional testing has been conducted by university researchers and nongovernmental agencies. Presence of residues needs to be put into the context of safety standards. Furthermore, to appropriately interpret residue data, analytical assays must be validated for each food sample matrix. Regulatory agency surveys indicate that 99% of glyphosate residues in food are below the European maximum residue limits (MRLs) or U.S. Environmental Protection Agency tolerances. These data support the conclusion that overall residues are not elevated above MRLs/tolerances due to agricultural practices or usage on genetically modified (GM) crops. However, it is important to understand that MRLs and tolerances are limits for legal pesticide usage. MRLs only provide health information when the sum of MRLs of all foods is compared to limits established by toxicology studies, such as the acceptable daily intake (ADI). Conclusions from dietary modeling that use actual food residues, or MRLs themselves, combined with consumption data indicate that dietary exposures to glyphosate are within established safe limits. Measurements of glyphosate in urine can also be used to estimate ingested glyphosate exposure, and studies indicate that exposure is

Klebsiella pneumoniae is a common cause of Gram-negative pneumonia. The spread of antibiotic-resistant and hypervirulent strains has made treatment more challenging. This study sought to determine the immunomodulatory, antibacterial, and therapeutic potential of purified murine stem cell Ag-1+ (Sca-1+) lung mesenchymal stem cells (LMSCs) using in vitro cell culture and an in vivo mouse model of pneumonia caused by K. pneumoniae. Sca-1+ LMSCs are plastic adherent, possess colony-forming capacity, express mesenchymal stem cell markers, differentiate into osteogenic and adipogenic lineages in vitro, and exhibit a high proliferative capacity. Further, these Sca-1+ LMSCs are morphologically similar to fibroblasts but differ ultrastructurally. Moreover, Sca-1+ LMSCs have the capacity to inhibit LPS-induced secretion of inflammatory cytokines by bone marrow–derived macrophages and neutrophils in vitro. Sca-1+ LMSCs inhibit the growth of K. pneumoniae more potently than do neutrophils. Sca-1+ LMSCs also possess the intrinsic ability to phagocytize and kill K. pneumoniae intracellularly. Whereas the induction of autophagy promotes bacterial replication, inhibition of autophagy enhances the intracellular clearance of K. pneumoniae in Sca-1+ LMSCs during the early time of infection. Adoptive transfer of Sca-1+ LMSCs in K. pneumoniae–infected mice improved survival, reduced inflammatory cells in bronchoalveolar lavage fluid, reduced inflammatory cytokine levels and pathological lesions in the lung, and enhanced bacterial clearance in the lung and in extrapulmonary organs. To our knowledge, these results together illustrate for the first time the protective role of LMSCs in bacterial pneumonia.

Objective Our objective was to evaluate subsequent feedlot performance and carcass characteristics of steers that previously grazed endophyte-infected fescue and received either a mineral only (MIN) supplement or a cumulative management (CM) strategy. Materials and Methods Grazing occurred in autumn and winter of 2018 and 2019 with feedlot finishing after grazing. Within year, 60 fall-weaned steers (282 ± 7.7 kg) were randomly assigned to either MIN (n = 3) or CM (n = 3) pastures. The CM strategy include a steroidal growth implant, monensin, and 1% BW (as-fed basis) of 1:1 corn gluten feed:soybean hull mix. During feedlot finishing, steers were penned to maintain pasture groups. Results and Discussion Steers receiving CM were heavier (P = 0.002) entering the feedlot and finished 40 d earlier than MIN (161 vs. 201 ± 5.7 d, P Implications and Applications A CM strategy increases weight of steers grazing toxic fescue. This additional weight can reduce the days required for feedlot finishing without influencing carcass characteristics. Improved grazing performance and fewer days for feedlot finishing may reduce environmental impact.

Centro de Investigación Forestal (CIFOR), Nonrecombining sex chromosomes, like the mammalian Y, often lose genes and accumulate transposable ele ments, a process termed degeneration. The correlation between suppressed recombination and degeneration is clear in animal XY systems, but the absence of recombination is confounded with other asymmetries between the X and Y. In contrast, UV sex chromosomes, like those found in bryophytes, experience symmetrical population genetic conditions. Here, we generate nearly gapless female and male chromosome-scale reference genomes of the moss Ceratodon purpureus to test for degeneration in the bryophyte UV sex chromosomes. We show that the moss sex chromosomes evolved over 300 million years ago and expanded via two chromosomal fusions. Although the sex chromosomes exhibit weaker purifying selection than autosomes, we find that suppressed recombination alone is insufficient to drive degeneration. Instead, the U and V sex chromosomes harbor thousands of broadly expressed genes, including numerous key regulators of sexual development across land plants., This work was supported by NSF DEB-1541005 and 1542609 and start-up funds from UF to S.F.M.; microMORPH Cross-Disciplinary Training Grant, Sigma-Xi Grant-In-Aid of Research, and Society for the Study of Evolution Rosemary Grant Award to S.B.C.; NSF DEB-1239992 to N.J.W.; the Emil Aaltonen Foundation and the University of Turku to S.O.; and NSF DEB-1541506 to J.G.B. and S.F.M. The work conducted by the U.S. Department of Energy Joint Genome Institute was supported by the Office of Science of the U.S. Department of Energy under contract no. DE-AC02-05CH11231., 12 Pág. Supplementary material for this article is available at http://advances.sciencemag.org/cgi/ content/full/7/27/eabh2488/DC1

Aims Emerging evidence suggests that remnant cholesterol (RC) promotes atherosclerotic cardiovascular disease (ASCVD). We aimed to estimate RC-related risk beyond low-density lipoprotein cholesterol (LDL-C) and apolipoprotein B (apoB) in patients without known ASCVD. Methods and results We pooled data from 17 532 ASCVD-free individuals from the Atherosclerosis Risk in Communities study (n = 9748), the Multi-Ethnic Study of Atherosclerosis (n = 3049), and the Coronary Artery Risk Development in Young Adults (n = 4735). RC was calculated as non-high-density lipoprotein cholesterol (non-HDL-C) minus calculated LDL-C. Adjusted Cox models were used to estimate the risk for incident ASCVD associated with log RC levels. We also performed discordance analyses examining relative ASCVD risk in RC vs. LDL-C discordant/concordant groups using difference in percentile units (>10 units) and clinically relevant LDL-C targets. The mean age of participants was 52.3 ± 17.9 years, 56.7% were women and 34% black. There were 2143 ASCVD events over the median follow-up of 18.7 years. After multivariable adjustment including LDL-C and apoB, log RC was associated with higher ASCVD risk [hazard ratio (HR) 1.65, 95% confidence interval (CI) 1.45–1.89]. Moreover, the discordant high RC/low LDL-C group, but not the low RC/high LDL-C group, was associated with increased ASCVD risk compared to the concordant group (HR 1.21, 95% CI 1.08–1.34). Similar results were shown when examining discordance across clinical cutpoints. Conclusions In ASCVD-free individuals, elevated RC levels were associated with ASCVD independent of traditional risk factors, LDL-C, and apoB levels. The mechanisms of RC association with ASCVD, surprisingly beyond apoB, and the potential value of targeted RC-lowering in primary prevention need to be further investigated.

Recent studies have highlighted the functional diversity of dermal fibroblast populations in health and disease, with part of this diversity linked to fibroblast lineage and embryonic origin. Fibroblasts derived from foxd1‐expressing progenitors contribute to the myofibroblast populations present in lung and kidney fibrosis in mice but have not been investigated in the context of dermal wound repair. Using a Cre/Lox system to genetically track populations derived from foxd1‐expressing progenitors, lineage‐positive fibroblasts were identified as a subset of the dermal fibroblast population. During development, lineage‐positive cells were most abundant within the dorsal embryonic tissues, contributing to the developing dermal fibroblast population, and remaining in this niche into adulthood. In adult mice, assessment of fibrosis‐related gene expression in lineage‐positive and lineage‐negative populations isolated from wounded and unwounded dorsal skin was performed, identifying an enrichment of transcripts associated with matrix synthesis and remodeling in the lineage‐positive populations. Using a novel excisional wound model, ventral skin healed with a greatly reduced frequency of foxd1 lineage‐positive cells. This work supports that the embryonic origin of fibroblasts is an important predictor of fibroblast function, but also highlights that within disparate regions, fibroblasts of different lineages likely undergo convergent differentiation contributing to phenotypic similarities.

The world acreage of organic crop production systems is increasing, but the soil microbial dynamics in these systems are not fully understood. We studied the composition and functioning of soil microbial communities in 4-year organic or conventional rotations. The organic systems were tilled to control weeds, and N2-fixing legumes or compost supplied nutrients. The conventional systems were managed under no-till; herbicides controlled weeds, and compost or fertilizers supplied nutrients. Soil microbial biomass C (MBC), the diversity and composition of bacterial communities, and activities of enzymes that mediate C, N, P, and S cycling were determined. The bacterial classes Gemmatimonadetes, C0119 (phylum Chloroflexi), and Thermomicrobia (phylum Chloroflexi) were more abundant in organic than conventional cropping systems, so were some genera from the class Actinobacteria. The bacterial β-diversity showed similar cropping system differences. However, acid phosphomonoesterase activity was greater in conventional than organic cropping systems, presumably because the soil P from the large amounts of compost applied in the organic system suppressed this enzyme. MBC, bacterial α-diversity, the relative abundances of the bacterial classes δ-Proteobacteria, γ-Proteobacteria, and Bacilli (phylum Firmicutes) were all greater in compost than no-compost treatments. The relative abundances of three genera from Actinobacteria class were negatively correlated with acid phosphomonoesterase activity due to the high relative abundances of these genera, but low acid phosphomonoesterase activities, where compost was applied in the organic system. Therefore, there were soil bacterial compositional differences between organic and conventional cropping systems, but only differences in the activities of a P cycling enzyme were detected.

Identifying environmental predictors of phenotype is fundamentally important to many ecological questions, from revealing broadscale ecological processes to predicting extinction risk. However, establishing robust environment—phenotype relationships is challenging, as powerful case studies require diverse clades which repeatedly undergo environmental transitions at multiple taxonomic scales. Actinopterygian fishes, with 32,000+ species, fulfil these criteria for the fundamental habitat divisions in water. With four datasets of body size (ranging 10,905–27,226 species), I reveal highly consistent size‐by‐habitat‐use patterns across nine scales of observation. Taxa in marine, marine‐brackish, euryhaline and freshwater‐brackish habitats possess larger mean sizes than freshwater relatives, and the largest mean sizes consistently emerge within marine‐brackish and euryhaline taxa. These findings align with the predictions of seven mechanisms thought to drive larger size by promoting additional trophic levels. However, mismatches between size and trophic‐level patterns highlight a role for additional mechanisms, and support for viable candidates is examined in 3439 comparisons., Establishing environmental predictors of body size is of fundamental importance to ecology. With data from over 27,000 species of fishes, I establish general size rules between the major aquatic habitat types, revealing that various marine‐influenced environments contain larger taxa than those in freshwaters. The findings align with the predictions of seven mechanisms thought to drive larger size by promoting additional trophic levels, yet I demonstrate a need for additional mechanisms, and support for viable candidates is examined in 3439 comparisons.

Infection with HIV-1 remains uncurable due to reservoirs of latently infected cells. Any potential cure for HIV will require a mechanism to identify and target these cells in vivo. We created a panel of Jurkat cell lines latently infected with the HIV DuoFlo virus to identify candidate biomarkers of latency. SWATH mass spectrometry was used to compare the membrane proteomes of one of the cell lines to parental Jurkat cells. Several candidate proteins with significantly altered expression were identified. The differential expression of several candidates was validated in multiple latently infected cell lines. Three factors (LAG-3, CD147,CD231) were altered across numerous cell lines, but the expression of most candidate biomarkers was variable. These results confirm that phenotypic differences in latently infected cells exists and identify additional novel biomarkers. The variable expression of biomarkers across different cell clones suggests universal antigen-based detection of latently infected cells may require a multiplex approach.

Key message Overexpression of Zea mays SOC gene promotes flowering, reduces plant height, and leads to no reduction in grain production per plant, suggesting enhanced yield potential, at least, through increasing planting density. Abstract MIKC-type MADS-box gene SUPPRESSOR OF OVEREXPRESSION OF CONSTANS 1 (SOC1) is an integrator conserved in the plant flowering pathway. In this study, the maize SOC1 (ZmSOC1) gene was cloned and overexpressed in transgenic maize Hi-II genotype. The T0 plants were backcrossed with nontransgenic inbred B73 to produce first generation backcross (BC1) seeds. Phenotyping of both transgenic and null segregant (NT) BC1 plants was conducted in three independent experiments. The BC1 transgenic plants showed new attributes such as increased vegetative growth, accelerated flowering time, reduced overall plant height, and increased grain weight. Second generation backcross (BC2) plants were evaluated in the field using two planting densities. Compared to BC2 NT plants, BC2 transgenic plants, were 12–18% shorter, flowered 5 days earlier, and showed no reduction in grain production per plant and an increase in fat, starch, and simple sugars in the grain. Transcriptome comparison in young leaves of 56-day-old BC1 plants revealed that the overexpressed ZmSOC1 resulted in 107 differentially expressed genes. The upregulated transcription factor DNA BINDING WITH ONE FINGER 5.4 (DOF5.4) was among the genes responsible for the reduced plant height. Modulating expression of SOC1 opens a new and effective approach to promote flowering and reduce plant height, which may have potential to enhance crop yield and improve grain quality.