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Societal Impact Statement Pulse crops, including dry pea, lentil, and chickpea, are rich sources of protein, low digestible carbohydrates, and micronutrients. With the increasing demand for plant‐based protein with gluten‐free and allergen‐free foods, pulse crops have become of global importance for meeting the nutritional demand of growing populations. Breeding for nutritional quality is becoming a bottleneck for most breeding programs globally due to the cost of these available tools. Therefore, low‐cost, high‐throughput phenotyping tools will be a focus of interest for the selection of elite germplasm for cultivar development and gene identification for pulse cultivar development. This publication explains the emerging and future trends of phenotyping tools that are feasible for pulse breeding and improving nutritional quality. Summary Precision agriculture tools based on spectroscopic and imaging techniques now contribute to high‐throughput phenotyping (HTP) pipelines for nutritional and agronomic traits to speed breeding and selection for cultivar development. Fourier transform mid‐infrared (FT‐MIR) spectroscopy has been a reliable HTP tool for macro nutritional traits in pulse crops. Hyperspectral, multispectral, and RGB (red‐green‐blue) imaging with unmanned aerial systems (UAVs) have been developed to measure agronomic traits for cereals, but these techniques have yet to be developed and validated for pulse crops. This review summarizes different phenotyping techniques applied to nutritional and agronomic traits for crop breeding and reviews applications of machine learning tools for optimizing HTP.

Caffeine is a plant secondary metabolite, commonly known for its bioactivity properties. This molecule increases microbial activity during anaerobic digestion. The aim of this study was to determine the effects of caffeine doses on the rumen fermentation profile and nutrient digestibility when continuous culture fermenters were fed a lactating cow’s diet. We hypothesize that adding caffeine doses into continuous culture fermenters with a rumen fluid inoculum will not affect anaerobic fermentation or nutrient utilization. Fermenters were fed twice a day (at 0800 and 2000 h) with an experimental diet of high-producing dairy cows (53.55 g/DM day; Forage:Concentrate ratio, F:C of 40:60). Four levels of caffeine (0 ppm, 50 ppm, 100 ppm, and 150 ppm) were added to the diets as a treatment. The experiment was arranged in a randomized complete block design. Two blocks of four fermenters were run in two replicated periods of ten days. Statistical analyses were conducted in SAS version 9.4 for Windows (SAS Institute Inc., Cary, NC, USA) using the GLIMMIX procedure. The addition of caffeine at a 50 ppm dose on continuous culture fermentation decreased the protozoal counts (Diplodinium spp.) (p = 0.03) and ammonia concentrations (p < 0.05). A treatment of 50 ppm of caffeine increased the DM, OM, and starch digestibility (p < 0.05). When caffeine doses increased further than 50 ppm, the OM, DM, and starch digestibility decreased linearly (p = 0.01). The total volatile fatty acids and fatty acid proportions were unaffected. However, the acetate-to-propionate ratio (A:P) tended to decrease linearly among treatments (p = 0.07). The means of pH measurements and maximum pH had a significantly linear decrease effect (p < 0.01). Caffeine may represent a potential rumen fermentation modifier for use in lactating cow diets.

A significant factor determining the value of soybean is protein content, in addition to its importance as an oilseed. Increasing the seed protein content in soybean will make it a more valuable food and feedstock and enhance its importance in the international market. Due to this reason, developing high-protein soybean varieties is gaining increased industrial interest. To ensure profitable production, the stability of the high-protein trait needs to be verified under environmental stresses. The objectives of this study were to test whether the high-protein soybean genotypes can maintain their high seed protein content under heat and drought stresses imposed during seed filling and to assess the changes in their oil composition under stress conditions. The changes in seed protein and oil contents and oil composition of the high-protein lines NLM09–77, N14-7017, and N16-9924 under heat and drought stresses were assessed and compared against those of a recently released high-yielding, high-protein variety Benning HP; elite cultivars NC-Dunphy, NC-Raleigh, and NC-Dilday; low-protein breeding lines N11–9228 and N09-2505; heat-tolerant line DS25-1; and heat-susceptible line DT97-4290. We found that the high-protein genotypes NLM09–77, N16–9924, N14-7017, and Benning HP maintained their high seed protein contents under heat and drought stresses (46–55 %). Among them, NLM09–77 and N14-7017 were unique with no apparent negative relationship between the seed protein and oil contents. In terms of oil composition, N14-7017, NLM09-77, and Benning HP were similar to the elite cultivars NC-Dunphy and NC-Dilday. Additionally, the high-protein genotypes N14-7017, NLM09-77, and Benning HP did not exhibit any yield drag (2993–3150 kg ha−1 vs 3289–3368 kg ha−1 for the elite cultivars). Taken together, our study demonstrates the usefulness of the advanced breeding lines NLM09–77 and N14-7017 and cultivar Benning HP for developing new varieties with enhanced nutritional value under climate change-associated stresses.

Societal Impact Statement The world is increasingly looking to plant‐based sources to meet its protein needs. Multiple factors are driving this progression, ranging from nutritional and ethical considerations to climate change and population growth. As a pulse crop, lentil is ideal to help meet this change in demand. However, plant‐based proteins have limiting amino acids and lower protein digestibility compared to animal‐based proteins. This research identifies genetic markers that can be used to accelerate breeding of protein quality traits in lentil to ultimately help meet the rising demand in high‐quality plant‐based protein and bolster global food and nutritional security. Summary Lentil (Lens culinaris Medik.) contains ~25% high‐quality protein in addition to high concentrations of prebiotic carbohydrates and micronutrients, such as folate, iron, zinc, and selenium. As animal‐based protein's economic and environmental costs rise, plant‐based protein sources, such as lentil, will become increasingly important to global food systems. Consequently, evaluating and targeting protein quality traits for genomic‐assisted breeding is a valuable objective for lentil breeding programs. A diversity panel of 183 breeding lines was analyzed for protein quality traits, including amino acids and protein digestibility. Genotyping‐by‐sequencing (GBS) data were used to assess population structure and conduct genome‐wide association studies (GWAS). Genes in local linkage disequilibrium (LD) with significant single nucleotide polymorphism (SNP) markers were identified and categorized by homology. Protein quality traits showed a wide range of variation. Repeatability estimates were low to moderate across traits. Twelve traits were strongly correlated with each other (r > .7). Admixture analysis identified six ancestral subpopulations, which also demonstrated clustering in principal component analysis. Ten different traits had significant SNP associations; two loci were shared across multiple traits. Twenty‐seven candidate genes, including glutathione S‐transferase, protease family, and gibberellin 2‐beta‐dioxygenase genes, were identified. This paper identifies SNP markers associated with lentil protein quality traits. Once validated, these SNPs could accelerate lentil protein quality breeding efforts. By targeting lentil's limiting amino acids (methionine and cysteine) and protein digestibility through marker‐assisted selection, the nutritional value of lentil's protein content could be increased without the need to alter total protein content.

ABSTRACT Dry pea (Pisum sativum L.) is a highly nutritious cool season food legume or pulse crop within the Fabaceae family that features high levels of protein (20%–25%), prebiotic carbohydrates, and a range of minerals and vitamins. Dry pea is cultivated globally in temperate climates and consumed as a whole food, snack, or protein powder. Dry pea is featured in plant‐based meat items such as the “beyond” branded plant‐based meats. Dry pea is an excellent candidate for plant‐based protein alternatives due to the high protein and low‐fat concentrations present in the mature seed, but improvements are still needed for more widespread use. Breeding efforts are ongoing to further improve dry pea proteins' quality, quantity, and digestibility through biofortification. Global dry pea germplasm contains a wide array of accessions that are vital for dry pea breeding efforts focused on developing cultivars enriched with the most bioavailable forms of plant‐based proteins. The objective of this review is to summarize prior research exploring the factors that contribute to the nutritional value of the dry pea—especially protein quality and quantity.

Abstract Soil color has historically been estimated visually, but inaccuracies with this method have been documented, including disagreement in evaluators, color blindness, and differences in physical color books due to weathering. As new technologies become available to soil scientists, a reliable digital method to determine soil color in the field could offer a solution to these inaccuracies. This research aimed to assess if visual estimates of soil matrix color from 111 soil horizons differed from digital measurements of Munsell soil color across three South Carolina regions. Visual estimates of soil color were conducted using Munsell color books. The XRite Capsure gave digital measurements in Munsell notation specific for soils. The absolute difference between hues was similar among assessment methods. Visual estimates of color value and chroma were statistically greater than digital measurements. However, color values and chromas were less than one chip different, suggesting no practical difference between the two assessment methods. Utilizing a digital soil color assessment tool can assist with training and validate human estimates.

Over half of global food waste occurs in retail and households, highlighting the importance of improving shelf life. Many factors affect shelf life, including storage temperature, harvest time, processing method, and packaging. Modified atmosphere packaging (MAP), such as nitrogen (N2), has been used to improve the shelf life of produce. This study aimed to determine the effect of nitrogen-enriched packaging on the shelf life of greenhouse- and garden-grown kale in raised beds outdoors. Five kale cultivars (Black Magic, Darkibor, Lacinato, Red Russian, and Westlander) were grown in the greenhouse, and four cultivars (Curly Roja, Dwarf Green Curled, Meadowlark, and Vates) outdoors in raised beds, harvested and stored in nitrogen-enriched or nonenriched bags for 6 days. Leaf water content, water activity, color, visual score, and texture were measured for each replicate. Nitrogen-enriched packaging did not affect the kale shelf life. Garden-grown kale exhibited significantly lower water activity (0.979 vs. 0.999) and noticeably lower water content (88.6 vs. 90.6%) than greenhouse-grown kale. The cultivars were significant for all factors tested except for texture on day 4 and change in light to dark (Δ L*) and blue to yellow (Δ b*) color ratios. At the end of the study, cultivars Darkibor and Curly Roja had the highest visual score (4.4) and texture (305 g), respectively. The high performance of ‘Darkibor’ and ‘Curly Roja’ in visual score and texture may be due to decreased (Δ L*) and (Δ b*) values and decreased water content and water activity values. Future breeding efforts focused on the shelf life of kale should use the top-performing cultivars identified in this study and a diverse genetic population tested in a range of environments to develop cultivars with a longer shelf life.

Societal Impact Statement Since its inception in the early 20th century, organic agriculture has grown increasingly popular due to its focus on a holistic, environmentally friendly approach to crop production. However, it is comparatively limited in biomass production, disease management, nutritional quality, and postharvest treatment compared to conventional agriculture. These challenges carry over into kale production, the majority of which is certified organic. This article reviews organic kale production, morphology, and shelf life, focusing on nutrition and plant breeding. It explores the shortcomings of organic output and potential areas of study to enhance shelf life in organic kale while maintaining nutritional quality. Summary Organic production has grown exponentially over the past few decades in both acreage and popularity worldwide. This review focuses specifically on kale produced in the USA. However, regulations limiting synthetic inputs leave organic produce at a disadvantage compared to conventional agriculture in terms of biomass, nutritional quality, disease management, and postharvest treatment. Organic agriculture requires significant improvements to be a viable means of production for a growing population. Kale (Brassica oleracea var. acephala) is a “nutritional powerhouse” leafy green vegetable. The high concentration of vitamins, minerals, and prebiotic carbohydrates in a low‐calorie food makes kale an important crop for combatting obesity‐related non‐communicable diseases. However, the short shelf life of organic kale and inevitable fresh food waste make developing new kale cultivars with increased shelf life essential. This review article aims to (a) review kale morphology, consumer preference, and production, and (b) review nutritional quality, its effect on shelf life, and current breeding efforts of kale. Future research could focus on developing a kale breeding pipeline following suitable kale germplasm selection adapted to organic agriculture with both superior shelf life and improved nutritional quality.

Abstract Lentil, a cool-season food legume, is rich in protein and micronutrients with a range of prebiotic carbohydrates, such as raffinose-family oligosaccharides (RFOs), fructooligosaccharides (FOSs), sugar alcohols (SAs), and resistant starch (RS), which contribute to lentil's health benefits. Beneficial microorganisms ferment prebiotic carbohydrates in the colon, which impart health benefits to the consumer. In addition, these carbohydrates are vital to lentil plant health associated with carbon transport, storage, and abiotic stress tolerance. Thus, lentil prebiotic carbohydrates are a potential nutritional breeding target for increasing crop resilience to climate change with increased global nutritional security. This study phenotyped a total of 143 accessions for prebiotic carbohydrates. A genome-wide association study (GWAS) was then performed to identify associated variants and neighboring candidate genes. All carbohydrates analyzed had broad-sense heritability estimates (H 2) ranging from 0.22 to 0.44, comparable to those reported in the literature. Concentration ranges corresponded to percent recommended daily allowances of 2–9% SAs, 7–31% RFOs, 51–111% RS, and 57–116% total prebiotic carbohydrates. Significant SNPs and associated genes were identified for numerous traits, including a galactosyltransferase (Lcu.2RBY.1g019390) known to aid in RFO synthesis. Further studies in multiple field locations are necessary. Yet, these findings suggest the potential for molecular-assisted breeding for prebiotic carbohydrates in lentil to support human health and crop resilience to increase global food security.

Clarireedia jacksonii sp. nov. formerly Sclerotinia homoeocarpa F.T. Bennett, one of the causal agents of dollar spot, is the most widespread pathogen in turfgrass systems. Dollar spot (DS) affects both cool- and warm-season grasses, during a wide range of environmental conditions. Field studies were conducted at Clemson University, Clemson, SC, on a creeping bentgrass [Agrostis stolonifera L. var. palustris (Huds) cv. Crenshaw] putting green for 2 consecutive years from August to October in year 1 and July to September in year 2. The objective of the studies was to evaluate biological control agents (BCAs) and synthetic fungicides at reduced rates for their efficacy controlling dollar spot. Four replications of 1.5 × 1.5-m plots were used in the experimental design. Treatments included the following: Bacillus subtilis (BS); plant extract oils (EO) including clove oil + wintergreen oil + thyme oil; extract of Reynoutria sachalinensis (RS); Bacillus licheniformis (BL); chlorothalonil (CL); and azoxystrobin + propiconazole (AzP). Synthetic fungicides were used at reduced rates in combination with biological control agents, to evaluate curative control efficacy of various combinations. All reduced synthetic programs, except CL + EO, provided acceptable disease severity (≤15%) at the end of year 1 and acceptable (≥7) turfgrass visual quality. Azoxystrobin + propiconazole, CL, AzP + BL, AzP + EO, AzP + BS all provided ≤15% disease severity and ≥7 visual turfgrass quality 14 days after the last application in year 2.

The objective of this study was to evaluate the yield, nutritional composition, and digestibility of conventional (CON) and brown midrib (BMR) pearl millet (PM) with different establishment dates, maturity at harvest and when mixed with cowpea (CWP). In trial 1, CON and BMR were planted on two different dates. In trial 2, CON and BMR, mixed or not with CWP, were harvested when PM was at the boot or heading stages. In trial 1, dry matter (DM) yield was similar between both PM genotypes but delaying establishment reduced DM yield by 30%. Additionally, BMR had a lower concentration of acid detergent lignin (ADL) and a higher in vitro neutral detergent fiber digestibility (IVNDFD) compared to CON. In Trial 2, the DM yield was 7.3% higher for CON compared to BMR, and PM with the BMR trait had a lower level of ADL and higher IVNDFD compared to CON. Mixing PM with CWP had negligible effects on nutritional composition but reduced DM yield by 8.3%. Results of these studies indicated that fiber from BMR PM is more digestible than CON but, in one of the trials, this occurred at the expense of lower DM yield. Mixing CWP with PM negatively impacted DM yield.

Chickpea is a highly nutritious pulse crop with low digestible carbohydrates (40–60%), protein (15–22%), essential fats (4–8%), and a range of minerals and vitamins. The fatty acid composition of the seed adds value because fats govern the texture, shelf-life, flavor, aroma, and nutritional composition of chickpea-based food products. Therefore, the biofortification of essential fatty acids has become a nutritional breeding target for chickpea crop improvement programs worldwide. This paper examines global chickpea production, focusing on plant lipids, their functions, and their benefits to human health. In addition, this paper also reviews the chemical analysis of essential fatty acids and possible breeding targets to enrich essential fatty acids in chickpea (Cicer arietinum) biofortification. Biofortification of chickpea for essential fatty acids within safe levels will improve human health and support food processing to retain the quality and flavor of chickpea-based food products. Essential fatty acid biofortification is possible by phenotyping diverse chickpea germplasm over suitable locations and years and identifying the candidate genes responsible for quantitative trait loci mapping using genome-wide association mapping.

Dry soil conditions at soybean planting results in poor stand establishment, which often necessitates replanting. We conducted a study to identify soybean genotypes that can maintain germination rates and possess better root morphology under water stress. We tested 41 Plant Introductions (PI) for germination and seedling root traits under controlled environmental conditions at five water potentials: 0.00, −0.27, −0.54, −0.82, and −1.09 MPa (no, low, mild, severe, and extreme water stress, respectively). The same genotypes were tested for emergence and seedling root traits under field conditions in South Carolina (2021 and 2022) and North Carolina (2022). Among the 41 genotypes evaluated, PI 398566 and PI 424605A maintained higher germination percentages (≥63%) under water stress. The same genotypes were ranked among the top 15 genotypes for root traits (total-root and fine-root (diameter between 0.25 and 0.50 mm) length, surface area, and/or volume) under water stress. Furthermore, they had relatively higher emergence percentages under field conditions (≥35% under dry soil conditions). The superior genotypes identified in this study (PI 398566 and PI 424605A) that had better germination and root morphology under water-stress and no-stress conditions and better emergence would be useful for developing varieties with drought tolerance during the emergence phase.

Musculoskeletal injuries can lead to a working dog being withdrawn from service prior to retirement. During training exercises, young working dogs are often required to perform repetitive tasks, including adoption of an upright posture (or “hupp” task). Non-invasive, quantitative methods would be helpful for supporting research on effects of these repetitive tasks on sacroiliac joints (SIJ). Furthering our understanding of lesions in and biomechanical stresses on the SIJ could provide insight into possible training modifications for minimizing risks of SIJ injury. Aims of this retrospective, secondary analysis, exploratory study were to test hypotheses that (1) mean numbers of SIJ computed tomographic (CT) lesions/dog would differ among work status groups in young working Labrador Retrievers; (2) a methodology for using CT data and finite element analysis (FEA) to quantify SIJ ligament strain in the static canine pelvis would be feasible; and (3) this FEA methodology would yield repeatable measures of SIJ ligament strain. Clinical and CT data for 22 Labrador retriever working dogs, aged 11–48 months, were retrospectively reviewed. Dogs were categorized into three work status groups (Breeder, Detection, Other). A veterinary radiologist who was unaware of dog group status recorded numbers of CT lesions for each SIJ, based on previously published criteria. Mean numbers of SIJ CT lesions/dog were compared among dog work status groups. An a priori FEA model was created from the CT images of one of the dogs using image analysis software packages. Using tissue properties previously published for the human pelvis, various directional loads (n = 8) and forces (48 ligament strain values) were placed on the canine model in five trials. Repeatability was tested using regression analysis. There was a significantly greater mean number of subchondral sclerosis lesions in left SIJ of Breeder vs. Detection dogs, a significantly greater mean number of subchondral cysts in right SIJ for Detection vs. Breeder dogs, and a significantly greater mean number of subchondral cysts in right SIJ of Other vs. Breeder dogs (p < 0.05). Finite element modeling and analysis using CT data was feasible and yielded repeatable results in 47/48 (98%) of tests at each combination of strain, ligament, and side.

Researchers who study the selection and breeding program criteria for military working dogs aim to help maximize the years of active duty service. Computed tomographic (CT) quantitative phenotyping has been previously described as a method for supporting these research studies. Funnel-shaped lumbar vertebral foramen malformations have been previously described in Labrador retriever military working dogs and proposed to be risk factors for impaired arterial perfusion of nerve tissues during exercise. Articular process dysplasia malformations have been previously described in varying dog breeds and proposed to be risk factors for articular process degenerative joint disease and vertebral foramen stenosis. Aims of this retrospective, cross-sectional study were to describe quantitative CT phenotyping methods for characterizing funnel-shaped lumbar vertebral foramina and articular process dysplasia malformations and to apply these methods in a comparison between groups of German shepherd and Belgian Malinois military working dogs. A military working dog hospital's database was searched for German shepherd and Belgian Malinois dogs aged

Regulatory affairs professionals play pivotal roles in ensuring healthcare products adhere to regulations and in gaining regulatory approval for product manufacture and sales. To do this, they must understand the science and technology connected with a product, the company's business goals, and, most importantly, the nuances of national and international regulations and guidances connected to the product. But although they perform complicated work connected to the entire product development lifecycle, surveys have indicated only 14% of regulatory professionals come to the field with a degree related to the work and for more than half, regulatory work is a “second career.” The net result is a heterogeneous professional population that must learn complex, detailed work on the fly in as short a time as possible. Without a structure to guide development, these expectations are a challenge for someone new to the field, that person's supervisor, and for training developers. Various non-profit groups have created competency models to provide this structure, but because competencies only identify traits demonstrated by high-performing professionals, not the specific tasks associated with individual roles, these models have had limited impact on the profession. Identifying and structuring actionable tasks based on a competency model would increase the model's utility, dissemination, and usage. Entrustable professional activities might provide the methodology for doing so.

[This corrects the article DOI: 10.1371/journal.pone.0163890.].

Endophyte-infected tall fescue (E+) produces ergovaline and ergovalinine, which are mycotoxins that act as dopamine agonists to suppress prolactin and induce vasoconstriction. The experiment was designed as a 3 × 2 × 2 factorial with DRD2 genotype (AA, AG, GG), fescue seed (endophyte-free, E− or endophyte-infected, E+), stage of gestation (MID, d (day) 35–85; LATE, d 86–parturition) and all interactions in the model. Pregnant Suffolk ewes (n = 60) were stratified by genotype and fed E+ or E− seed in a total mixed ration according to treatment assignment. Serum prolactin concentrations were lower (p < 0.05) in ewes fed E+ seed but did not differ by maternal DRD2 genotype or two-way interaction. Lamb birth weight was lower (p < 0.05) in ewes fed E+ seed in last trimester. Pre-weaning growth rate, milk production and total weaning weight was reduced (p < 0.05) in ewes fed E+ fescue seed during MID and LATE gestation. Ingestion of ergovaline/ergovalinine in last trimester reduces lamb birth weight; however, lamb growth rate, milk production and total weaning weight are reduced in all ewes fed E+ during mid and last trimester.

Weaned lambs (n = 82), born to ewes fed endophyte-free (E−) or endophyte-infected (E+; 1.77 mg hd−1 d−1 ergovaline + ergovalinine) tall fescue seed from d 35 to 85 of gestation (MID) and/or d 86 of gestation to parturition (LATE), were used to examine how ergot alkaloid exposure during fetal development altered subsequent puberty attainment or carcass quality. Lambs were weaned at 75 d of age and separated by sex to assess puberty in ewe lambs (n = 39) and to evaluate growth, carcass and meat quality in wethers (n = 43). Data were analyzed with maternal fescue treatment, stage of gestation, and two-way interaction in the model. Age at puberty tended (P = 0.06) to be longer for ewe lambs born to dams fed E+ fescue during LATE gestation versus those fed E−. Post-weaning average daily gain tended to be higher (P = 0.07) for wether lambs born to dams fed E+ fescue seed during MID gestation compared to E−. Exposure to ergot alkaloids during fetal growth altered (P < 0.10) longissimus muscle weight and color, lipid deposition, fatty acid composition, and shear force values of semimembranosus muscle in wether lambs. These results indicate that exposure to ergot alkaloids in utero does alter subsequent post-weaning puberty attainment and body composition in offspring.

Recreation-based health and wellness programs for military service members are currently a topic of significant interest in the recreation and health industries. This study examined the health outcomes associated with participation in Project Sanctuary, a week-long recreation-based health and wellness family retreat for injured military service members. Linear mixed modeling was used to examine changes in health outcomes over four time points, and considered multiple covariates. One-hundred twenty-seven service members participated. Statistically significant reductions in total scores for Post-Traumatic Stress Disorder Checklist (PCL) measures were found, as well as notable improvements in Depression, Anxiety, and Atress Scale (DASS) and mental health functioning immediately following the intervention. No changes were found in physical health functioning. Trends demonstrated that participants maintained the positive psychological health changes over the three and six month time points. Veteran's Administration (VA) disability rate was the only significant covariate associated with health outcome change across timepoints. Implications for future practice and research are discussed in the article.

Nodulation is energetically costly to the host: legumes balance the nitrogen demand with the energy expense by limiting the number of nodules through long-distance signaling. A split root system was used to investigate systemic autoregulation of nodulation (AON) in Medicago truncatula and the role of the AON genes RDN1 and SUNN in the regulatory circuit. Developing nodule primordia did not trigger AON in plants carrying mutations in RDN1 and SUNN genes, while wild type plants had fully induced AON within three days. However, despite lacking an early suppression response, AON mutants suppressed nodulation when roots were inoculated 10 days or more apart, correlated with the maturation of nitrogen fixing nodules. In addition to correlation between nitrogen fixation and suppression of nodulation, suppression by extreme nutrient stress was also observed in all genotypes and may be a component of the observed response due to the conditions of the assay. These results suggest there is more than one systemic regulatory circuit controlling nodulation in M. truncatula. While both signals are present in wild type plants, the second signal can only be observed in plants lacking the early repression (AON mutants). RDN1 and SUNN are not essential for response to the later signal.

Indirect calorimetry (IC) measurements to estimate resting energy expenditure (REE) necessitate a stable measurement period, or steady state (SS). There is limited evidence when assessing the time to reach SS in young, healthy adults. The aims of this prospective study are to determine the approximate time to necessary reach SS using open-circuit IC and to establish the appropriate duration of SS needed to estimate REE. One hundred young, healthy participants (54 males and 46 females; age = 20.6 ± 2.1 years; body weight = 73.6 ± 16.3 kg; height 172.5 ± 9.3 cm; BMI = 24.5 ± 3.8 kg/m2) completed IC measurement for approximately 30-minutes while the volume of oxygen (VO2) and volume of carbon dioxide (VCO2) were collected. SS was defined by variations in the VO2 and VCO2 of ≤10% coefficient of variation (%CV) over a period of 5- consecutive minutes. The 30-minute IC measurement was divided into six 5-minute segments, S1, S2, S3, S4, S5 and S6. The results show that SS was achieved during S2 (%CV = 6.81 ± 3.2%), and the %CV continued to met the SS criteria for the duration of the IC measurement (S3= 8.07 ± 4.4%; S4 = 7.93 ± 3.7%; S5 = 7.75 ± 4.1%; S6 = 8.60 ± 4.6%). The current study found that in a population of young, healthy adults the duration of the IC measurement period could be a minimum of 10 minutes. The first 5-minute segment was discarded, while SS occurred by the second 5-minute segment.

Thrombocytes are nucleated platelets involved in immune functions such as pathogen recognition and release of pro-inflammatory bioactive compounds when exposed to bacterial and viral molecules. However, the complete role of these cells in innate and adaptive immune responses is not understood, and little is known about their biology at the molecular-genetic level. Highly sensitive RNA-sequencing technologies were used to analyze the complete transcriptome of thrombocytes for the first time with analytical resolution focused on cell-based components of the immune system/response. Amongst all the genes listed in the current chicken genome assembly, 10,041 gene transcripts were found in the chicken thrombocyte. After 1-hour in vitro stimulation with lipopolysaccharide (LPS, Salmonella minnesota), 490 genes were upregulated and 359 genes were downregulated, respectively, with at least a 1-fold change relative to unexposed thrombocytes. Additionally, by constructing a de novo assembly, we were able to identify a total of 3,030 novel genes in the thrombocyte transcriptome. The information generated here is useful in development of novel solutions to lower the economic burden and zoonotic threat that accompanies infectious diseases for birds and fish. In addition, the resources created here have translational utility as a model system to find orthologous genes and genes related to its enucleated counterpart, the platelet.