A Translational Approach to Increase Pulse Intake and Promote Public Health through Developing an Extension Bean Toolkit.

Practical, affordable solutions need to be implemented to address global challenges confronting human and environmental health. Despite a myriad of benefits for people and the planet, beans and other pulses (e.g., chickpeas, cowpeas, dry peas, lentils) are under-consumed. To better understand consumer concerns and interests, a Food Habits Survey was conducted and the findings were incorporated into the Colorado State University Extension Bean Toolkit. Guided by the Information-Motivation-Behavioral Skills model, the toolkit included informational social media posts, cooking guidance, and an online class. A convenience sample of participants was recruited through Extension and university networks. After class participation, significant gains in knowledge of pulse nutrition, versatility, and cooking were observed, with an average increase of 1.5 points on a 5-point Likert scale (p < 0.001). Moreover, participants (n = 86) perceived a greater importance of motivators (e.g., nutrition, versatility, environmental benefits) and found barriers (e.g., flatulence, long cooking times, unfamiliarity) to be less discouraging. Most participants reported an intention to eat more pulses, and among those who completed the 1-month follow-up survey, pulse intake frequency increased (p = 0.004). Emphasizing motivating factors while simultaneously mitigating barriers to consumption can help reverse insufficient intake and promote healthy behavior change. Leveraging Extension or similar networks is one way to adopt a translational approach to better reach the public with this information.

Bean Cuisine: The Potential of Citizen Science to Help Motivate Changes in Pulse Knowledge and Consumption.

Pulses, or the dry, edible seeds of non-oilseed legumes (e.g., chickpeas, cowpeas, dry beans, dry peas, and lentils), are uniquely positioned to simultaneously benefit human and environmental well-being, all while being affordable and important to diverse cultural food traditions around the world. Despite the benefits they can provide, pulses are dramatically under-consumed. One key barrier preventing higher intake among consumers is a lack of familiarity with how to prepare and regularly incorporate pulses into meals. To address this barrier and actualize findings from our laboratory, we created the Bean Cuisine, a 2-week cuisine (i.e., meal plan) with 56 pulse-centric recipes corresponding to 14 unique breakfast, lunch, snack, and dinner ideas. Each meal category was largely interchangeable, i.e., the order of the breakfast recipes is not important, and one could be swapped for another if a different order were preferrable to a consumer. Fifty-six citizen scientists were recruited to provide feedback on the Bean Cuisine. Free response feedback related to project participation was very positive, and common themes included changes in pulse consumption and cooking behaviors, increased awareness of pulse variety and versatility, and positive perceptions of citizen science. Overall, participation in the Bean Cuisine citizen science project helped create pulse advocates, empowering participants to advance the well-being of their communities through pulses.

Defining Nutritional and Functional Niches of Legumes: A Call for Clarity to Distinguish a Future Role for Pulses in the Dietary Guidelines for Americans.

Legume food crops can contribute to the solution of diet-related public health challenges. The rich diversity of the botanical family Fabaceae (Leguminosae) allows legumes to fill numerous nutritional niches. Pulses (i.e., a subgroup of legumes including chickpeas, cowpeas, dry beans, dry peas, and lentils) are a nutrient-dense food that could play a key role in eliminating the dramatic underconsumption of dietary fiber and potassium, two dietary components of public health concern, all while maintaining a caloric intake that promotes a healthy weight status. However, incorrect use of terminology-in the commercial and scientific literature as well as in publications and materials prepared for the consuming public-creates confusion and represents a barrier to dissemination of clear dietary guideline messaging. The use of accurate terminology and a simple classification scheme can promote public health through differentiation among types of legumes, better informing the development and implementation of nutritional policies and allowing health care professionals and the public to capitalize on the health benefits associated with different legumes. Although inconsistent grouping of legumes exists across countries, the recently released 2020-2025 Dietary Guidelines for Americans (DGA) were chosen to illustrate potential challenges faced and areas for clarification. In the 2020-2025 DGA, pulses are included in two food groups: the protein food group and 'beans, peas, lentils' vegetable subgroup. To evaluate the potential of pulses to contribute to intake of key dietary components within calorie recommendations, we compared 100 kilocalorie edible portions of pulses versus other foods. These comparisons demonstrate the unique nutritional profile of pulses and the opportunity afforded by this type of legume to address public health concerns, which can be greatly advanced by reducing confusion through global harmonization of terminology.

Motivating Pulse-Centric Eating Patterns to Benefit Human and Environmental Well-Being.

Pulses (e.g., lentil, common bean, chickpea, and dry pea) are linked to a myriad of positive human and environmental health impacts, making them an ideal food for wise and conscientious global citizens. In addition, pulses are affordable and shelf-stable. The combination of these factors, an elevated consumer interest in plant-based diets, and the COVID-19 pandemic resulted in increased purchasing of pulses and even empty grocery store shelves. Although pulses have many associated benefits, some consumers are hesitant to regularly eat pulses, claiming concerns of abdominal discomfort or a lack of knowledge on how to best prepare pulses. To capitalize on increased consumer interest and purchasing of pulses, now is the time for outreach efforts that address these concerns and the positive outcomes associated with pulses, thereby promoting public and environmental health. Consumers must actively decide to add pulses to their grocery lists and incorporate them into their regular eating patterns. Motivation to adopt new eating habits is essential because knowledge alone does not result in behavior change. Thus, to mitigate perceived barriers and drive consumption, we suggest application of the Information-Motivation-Behavioral Skills Model and emphasis of three main benefits of pulses as motivators: (1) culinary versatility, (2) sustainability, and (3) healthfulness.

Production of a recombinant monoclonal antibody to Herpes Simplex Virus glycoprotein D for immunoaffinity purification of tagged proteins.

We have developed a stable Chinese Hamster Ovary (CHO) cell line for the production of a recombinant monoclonal antibody (mAb) to a short protein sequence derived from the N-terminus of human herpes simplex virus type 1 glycoprotein D (HSV-1 gD). The antibody (designated r34.1) provides a useful tool for the immunoaffinity purification of HSV-1 gD tagged proteins, and provides a generic purification system by which various proteins and peptides can be purified. Recombinant 34.1 was assembled using cDNA derived from a HSV-1 gD specific murine hybridoma engineered to encode a full-length IgG molecule. Antibody expression cassettes were transfected into CHO-S cells, and a stable cell-line expressing up to 500 mg/L of antibody, isolated. Affinity purified r34.1 exhibited nanomolar affinity for its cognate ligand, and is stable throughout multiple cycles of immunoaffinity purification involving ligand binding at neutral pH, followed by acid elution. The HSV-1 gD tag expression and purification strategy has been used to enhance the secretion and purification of several vaccine immunogens including HIV envelope protein rgp120s, but the protocol has potential for generic application.

Characterization of MHC class IA in the endangered southern corroboree frog.

Southern corroboree frogs (Pseudophryne corroboree) have declined to near extinction in the wild after the emergence of the amphibian chytrid fungus Batrachochytrium dendrobatidis in southeastern Australia in the 1980s. A major captive breeding and reintroduction program is underway to preserve this iconic species, but improving resistance to B. dendrobatidis would help the wild population to be self-sustaining. Using 3' and 5' rapid amplification of complementary DNA ends (RACE), we characterized the major histocompatibility complex (MHC) class IA locus in this species. We then used sequences generated from RACE to design primers to amplify the peptide-binding region (PBR) of this functional genetic marker. Finally, we analysed the diversity, phylogeny, and selection patterns of PBR sequences from four P. corroboree populations and compared this with other amphibian species. We found moderately high MHC class IA genetic diversity in this species and evidence of strong positive and purifying selection at sites that are associated with putative PBR pockets in other species, indicating that this gene region may be under selection for resistance to Bd. Future studies should focus on identifying alleles associated with Bd resistance in P. corroboree by performing a Bd laboratory challenge study to confirm the functional importance of our genetic findings and explore their use in artificial selection or genetic engineering to increase resistance to chytridiomycosis.

Complete mitochondrial genome of Dryophytes suweonensis (Anura Hylidae).

Dryophytes suweonensis is an endangered species with fragmented and declining populations from the Korean peninsula. We described 17,448 bp of D. suweonensis mtDNA, which had a shorter D-loop than other closely related species. The variation in nucleotide composition was similar to that of Hyla tsinlingensis but was larger than the one of its sister clade, D. japonicus.

Multiple major histocompatibility complex class I genes in Asian anurans: Ontogeny and phylogeny.

Amphibians, as the first terrestrial vertebrates, offer a window into early major histocompatibility complex (MHC) evolution. We characterized the MHC class I of two Korean amphibians, the Asiatic toad (Bufo gargarizans) and the Japanese tree frog (Hyla japonica). We found at least four transcribed MHC class I (MHC I) loci, the highest number confirmed in any anuran to date. Furthermore, we identified MHC I transcripts in terrestrial adults, and possibly in aquatic larvae, of both species. We conducted a phylogenetic analysis based on MHC I sequence data and found that B. gargarizans and H. japonica cluster together in the superfamily Nobleobatrachia. We further identified three supertypes shared by the two species. Our results reveal substantial variation in the number of MHC I loci in anurans and suggest that certain supertypes have particular physiochemical properties that may confer pathogen resistance.

Major histocompatibility complex selection dynamics in pathogen-infected túngara frog (Physalaemus pustulosus) populations.

Pathogen-driven selection can favour major histocompatibility complex (MHC) alleles that confer immunological resistance to specific diseases. However, strong directional selection should deplete genetic variation necessary for robust immune function in the absence of balancing selection or challenges presented by other pathogens. We examined selection dynamics at one MHC class II (MHC-II) locus across Panamanian populations of the túngara frog, Physalaemus pustulosus, infected by the amphibian chytrid fungus Batrachochytrium dendrobatidis (Bd). We compared MHC-II diversity in highland túngara frog populations, where amphibian communities have experienced declines owing to Bd, with those in the lowland region that have shown no evidence of decline. Highland region frogs had MHC variants that confer resistance to Bd. Variant fixation appeared to occur by directional selection rather than inbreeding, as overall genetic variation persisted in populations. In Bd-infected lowland sites, however, selective advantage may accrue to individuals with only one Bd-resistance allele, which were more frequent. Environmental conditions in lowlands should be less favourable for Bd infection, which may reduce selection for specific Bd resistance in hosts. Our results suggest that MHC selection dynamics fluctuate in túngara frog populations as a function of the favourability of habitat to pathogen spread and the vulnerability of hosts to infection.