Genotype Main Effect Research Articles

Thermoregulatory Response, Growth Performance and Organ Weights of FUNAAB-Alpha Chicken Genotypes Administered Sweet Citrus Peel Powder.

Thermoregulatory response and growth performance of FUNAAB-Alpha chickens administered sweet citrus peel powder (SCPP) were investigated in a twelve weeks experiment. A total of 360 1-day-old FUNAAB Alpha chicks were used for the study, comprising 120 birds per genotype: Normal feather (NF), Naked neck (NN) and Frizzle feather (FF). Birds were randomly allotted into 12 treatment groups: NF: 0 g, 3 g, 6 g and 9 g of SCPP, NN: 0 g, 3 g, 6 g and 9 g of SCPP, FF: 0 g, 3 g, 6 g and 9 g of SCPP. Each treatment was replicated thrice with 10 birds per replicate. No (p > 0.05) interactive and main effect of genotype and dosage of SCPP was observed on the Rectal temperature, Skin temperatures under wing and of the breast among the treatments. Similar (p > 0.05) interactive effect was observed on the respiratory rate (RR), however, higher (p < 0.001) RR was recorded in NN and FF compared with NF. Birds in the control group had the highest (p < 0.005) RR compared with birds administered 3 g and 9 g of SCPP/L of water. Interaction and main effect of SCPP dosage on heterophil (H), lymphocyte (L) and heterophil lymphocyte ratio (H/L) were not significant (p > 0.05) across the treatment groups. However, genotype significantly (p < 0.000) influenced H, L and H/L. The FF and NN had significantly (p < 0.000) lower H/L compared with the NF. Higher (p < 0.000) weight gain (WG) was observed in FF and NN compared to NF chickens. Significantly (p < 0.020) lower FCR was observed in 6 g/L (2.26), 3 g/L (2.65) and 9 g/L (2.66) of FF compared with NF on 9 g/L (4.03). Dosage of SCPP significantly (p < 0.001) influenced the relative weights of the intestine. In conclusion, lower H/L, higher WG and better FCR were observed in the NN and FF genotypes with the best FCR recorded at 6 g SCPP/L of water.

The Association of SLC16A1 (MCT1) Gene Polymorphism with Body Composition Changes during Weight Loss Interventions: A Randomized Trial with Sex-Dependent Analysis.

Monocarboxylates, transported by monocarboxylate transporters (MCTs), have been proposed to influence energy homeostasis and exhibit altered metabolism during exercise. This study investigated the association between the Asp490Glu (T1470A) (rs1049434) polymorphism of the SLC16A1 (MCT1) gene and changes in body composition in males and females with overweight or obesity. The 173 participants (56.6% females) completed a 6-month randomized controlled trial, being assigned to the Supervised exercise group (strength, endurance, or strength plus endurance training) or the Physical Activity (PA) Recommendations group. Participants were genotypically categorized as Asp/Asp (TT), Asp/Glu (TA), or Glu/Glu (AA). ANCOVA analysis showed main effects for genotype and interaction genotype x group (p<0.05) for several variables, especially in females. Asp/Asp (TT) females showed greater reductions in total fat mass (-29.30±8.75% change), percentage of body fat (-17.50±8.06% change), and body weight (-13.90±2.62% change) compared with Glu/Glu (AA) (p≤0.020; d≥0.18) and Asp/Glu (TA) (p≤0.050; d≥0.16) females, but only within the PA Recommendations group. Data indicated higher decreases in lean body mass of the Asp/Asp (TT) compared to Glu/Glu (AA) in females (p=0.067, η2p=0.067) and to Asp/Glu (TA) in males (p=0.026; η2p=0.101). Our investigation suggests an association of the SNP with body composition changes especially in females following exercise recommendations, highlighting the importance of supervised training to offset genetic predisposition for different weight loss. Differences in males only appeared for lean body mass, suggesting sex differences for this genetic association. The study underscores the potential influence of MCT1 functionality on human adaptations to weight loss interventions. ClinicalTrials.gov registration number, NCT01116856.

Evaluation and identification of high yielding and stable cowpea genotypes using GGE biplot and joint regression analysis for varietal development in East and Southern Africa

Malawi, Mozambique, and Tanzania's cowpea breeding programs have been collaborating on pre-breeding trials in different locations, focusing on key traits such as grain yield. However, there is a lack of information regarding the nature of genotype-environment interaction patterns among these environments. The study was conducted to assess and identify high-yielding and stable cowpea genotypes for the development of improved varieties in these three countries. Thirty genotypes were evaluated in ten environments using an incomplete block design with three replications. Grain yield data was analyzed using genotype (G) × genotype-environment interaction (GGE) biplot and joint regression analysis. The joint analysis of variance showed highly significant differences (P < 0.001) for the main effects of genotypes and environment, as well as their interaction effects. This suggested that the environments assessed were distinct and that the genotypes exhibited varying performances in response to environmental variations. The environmental indices showed that only four environments were favorable for high grain yield, and overall, Guariba was the topmost yielder (1973.5 kg ha−1) across all environments. The analysis identified IT10K-817-7 and MWcp08 as superior and stable genotypes, making them highly desirable and potentially recommended across test environments for their expected ability to achieve economically viable yields. Overall, Chitedze Research Station in Malawi and Nametil Research Station in Mozambique were both discriminating and representative, indicating that they are good test environments for selecting best-performing genotypes that can perform well across a wide range of conditions. These environments can be recommended as the best environments for future cowpea genotype evaluations targeting the release of varieties in the three countries (Malawi, Mozambique and Tanzania) where the METs were carried out.

Screening and Site Adaptability Evaluation of Qi-Nan Clones (Aquilaria sinensis) in Southern China

In recent years, plantations of Aquilaria sinensis in China have been dominated by Qi-nan, yet there remains limited research on the growth evaluation and breeding of these clones. In this study, a multi-point joint variance analysis, an additive main effect and multiplicative interaction (AMMI) model, a weighted average of absolute scores (WAASB) stability index, and a genotype main effect plus a genotype-by-environment interaction (GGE) biplot were used to comprehensively analyze the yield, stability, and suitable environment of 25 3-year-old Qi-Nan clones from five sites in southern China. The results showed that all the growth traits exhibited significant differences in the clones, test sites, and interactions between the clones and test sites. The phenotypic variation coefficient (PCV) and genetic variation coefficient (GCV) of the clones’ growth traits at the different sites ranged from 16.56% to 32.09% and 5.24% to 27.06%, respectively, showing moderate variation. The medium–high repeatability (R) of tree height and ground diameter ranged from 0.50 to 0.96 and 0.69 to 0.98, respectively. Among the clones, Clones G04, G05, G10, G11 and G13 showed good growth performance and could be good candidates for breeding. Environmental effects were found to be the primary source of variation, with temperature and light primarily affecting growth, while rainfall influenced survival and preservation rates. Yangjiang (YJ) was found to be the most suitable experimental site for screening high-yield and stable clones across the different sites, whereas the tree height and ground diameter at the Chengmai (CM) site were significantly higher than at the other sites, and the Pingxiang (PX) and Zhangzhou (ZZ) sites showed poor growth performance. The findings suggest that Qi-nan clones are suitable for planting in southern China. There were also abundant genetic variations in germplasm resources for the Qi-nan clones. The five selected clones could be suitable for extensive planting. Therefore, large-scale testing is necessary for determining genetic improvements in Qi-nan clones, which will be conducive to the precise localization of their promotion areas.

APOE×BDNF Interaction and Poorer Cognitive Outcomes Among Veterans With Mild Traumatic Brain Injury: An Exploratory Study.

The authors examined the interaction between apolipoprotein E (APOE) ε4 and brain-derived neurotrophic factor (BDNF) Val66Met alleles on neuropsychological functioning among veterans with histories of mild traumatic brain injury (mTBI). Participants were 78 veterans with mTBI (85% males; mean±SD age=32.95±7.00 years; mean time since injury=67.97±34.98 months) who completed a structured clinical interview and underwent a comprehensive neuropsychological assessment. Participants also provided a buccal swab for determination of their APOE and BDNF genotypes. Three cognitive composite scores were calculated from the neuropsychological assessment, reflecting visuospatial speed (seven variables), executive functioning (10 variables), and memory (eight variables). Two-way analyses of covariance (ANCOVAs) adjusted for age, sex, and race-ethnicity were used to assess the effects of APOE (ε4+ vs. ε4-) and BDNF (Met+ vs. Met-) on cognitive functioning. ANCOVAs revealed no significant main effects of APOE or BDNF genotypes on cognitive functioning; however, there was a significant APOE-by-BDNF genotype interaction for all three cognitive composite measures (visuospatial speed: ηp2=0.055; executive functioning: ηp2=0.064; and memory: ηp2=0.068). Specifically, the ε4+/Met+ (N=8) subgroup demonstrated the poorest cognitive functioning relative to all other allele subgroups (ε4+/Met-: N=12, ε4-/Met+: N=23, and ε4-/Met-: N=35). This exploratory study is the first to show that, compared with other allele subgroups assessed, veterans with both ε4 and Met alleles demonstrated the poorest cognitive functioning across several cognitive domains known to be negatively affected in the context of mTBI. Further research with larger sample sizes is needed to replicate these findings.

The Association of Genetic Markers Involved in Muscle Performance Responding to Lactate Levels during Physical Exercise Therapy by Nordic Walking in Patients with Long COVID Syndrome: A Nonrandomized Controlled Pilot Study.

Several genetic markers have shown associations with muscle performance and physical abilities, but the response to exercise therapy is still unknown. The aim of this study was to test the response of patients with long COVID through an aerobic physical therapy strategy by the Nordic walking program and how several genetic polymorphisms involved in muscle performance influence physical capabilities. Using a nonrandomized controlled pilot study, 29 patients who previously suffered from COVID-19 (long COVID = 13, COVID-19 = 16) performed a Nordic walking exercise therapy program for 12 sessions. The influence of the ACE (rs4646994), ACTN3 (rs1815739), AMPD1 (rs17602729), CKM (rs8111989), and MLCK (rs2849757 and rs2700352) polymorphisms, genotyped by using single nucleotide primer extension (SNPE) in lactic acid concentration was established with a three-way ANOVA (group × genotype × sessions). For ACE polymorphism, the main effect was lactic acid (p = 0.019). In ACTN3 polymorphism, there were no main effects of lactic acid, group, or genotype. However, the posthoc analysis revealed that, in comparison with nonlong COVID, long COVID increased lactic acid concentrations in Nordic walking sessions in CT and TT genotypes (all p < 0.05). For AMPD1 polymorphism, there were main effects of lactic acid, group, or genotype and lactic acid × genotype or lactic acid × group × genotype interactions (all p < 0.05). The posthoc analysis revealed that, in comparison with nonlong COVID, long COVID increased lactic acid concentrations in Nordic walking sessions in CC and CT genotypes (all p < 0.05). Physical therapy strategy through Nordic walking enhanced physical capabilities during aerobic exercise in post-COVID19 patients with different genotypes in ACTN3 c.1729C>T and AMPD1 c.34C>T polymorphisms. These findings suggest that individuals who reported long COVID who presumably exercised less beforehand appeared to be less able to exercise, based on lactate levels, and the effect of aerobic physical exercise enhanced physical capabilities conditioned by several genetic markers in long COVID patients.

Stability and genetic parameters for cassava yield attributes in the tropical humid region of Brazil

The performance differences in cassava genotypes arising from genotype vs. environment interactions (G × E) often lead to responses that are significantly lower than expected for selection. The objective of this study was to evaluate different stability methods, both parametric and non-parametric, such as additive main-effects and multiplicative interaction (AMMI), main effect of genotypes plus G × E (GGE), and weighted average of absolute scores (WAASB), in order to quantify the G × E in multi-environmental trials. A total of 12 genotypes were assessed across 12 environments using a completely randomized block design, with three replicates for traits such as fresh root yield (FRY) and dry matter content in the roots (DMC). The data were subjected to analysis of variance and the Scott Knott test (p < 0.05). The sum of squares (SQ) of genotypes, environment, and G × E effects were equally distributed for FRY, whereas for DMC, these effects accounted for 64.1%, 21.9%, and 13.8% of the SQ, respectively, indicating a lower environmental effect on this characteristic. Using the AMMI, GGE, and WAASB methods, genotypes with high agronomic performance and stability for FRY (BR11-34–41 and BR11-34–69) (> 32 t ha−1) and DMC (BRS Novo Horizonte, BR12-107–002, and BR11-24–156) (> 37%) were identified. The broad-sense heritability (h2\\documentclass[12pt]{minimal} \\usepackage{amsmath} \\usepackage{wasysym} \\usepackage{amsfonts} \\usepackage{amssymb} \\usepackage{amsbsy} \\usepackage{mathrsfs} \\usepackage{upgreek} \\setlength{\\oddsidemargin}{-69pt} \\begin{document}$${h}^{2}$$\\end{document}) for FRY and DMC was estimated to be 0.45 and 0.75, respectively. Approximately 72% of the methods identified BRS Novo Horizonte as the genotype with the highest stability and performance for DMC, while 47% identified genotypes BR11-34–41 and BR11-34–69 for FRY and intermediate DMC. Genotype BR11-24–156 exhibited high static stability according to 50% of the methods. Significant correlations were observed between stability and agronomic performance across the different methods, enabling the formation of groups based on stability concepts. Additionally, it was found that two mega-environments existed for FRY, whereas DMC displayed a single mega-environment with similar patterns, indicating an absence of G × E. We identified superior genotypes that could be promoted to national performance trials to develop stable cultivars with better yield attributes in cassava.

Exploring fatty acid composition, bioactive compounds, and antioxidant properties in oils of newly developed sesame mutant lines in Morocco

AbstractSesame (Sesamum indicum L.) is renowned for its significance as a global oilseed crop, valued for its high oil content and nutritional properties. This study aimed to explore the fatty acid composition, bioactive compounds, and antioxidant attributes of sesame mutant oils, examining their potential applications in food industry. Nine M4 mutants, along with their parents, were grown in two contrasting environments (Taoujdate and Afourare) to investigate their seed oil, phenolic and flavonoid contents, the balance between omega‐6, omega‐9, and omega‐3 fatty acids, iodine value, and susceptibility to oxidation. Genotype and environment main effects as well as their interaction had a significant effect on the majority of the parameters studied. The mutant “US2‐1” stands out with a distinctive fatty acid profile, featuring the highest saturated fatty acids, ω‐6/ω‐9 ratio, and omega‐3 content, in addition to its high total flavonoid content and remarkable oxidative stability. “ML2‐10,” “US1‐2,” “US2‐6,” and “US2‐7” exhibit high oil content, coupled with substantial levels of phenols and flavonoids, rendering them excellent candidates for culinary and industrial applications where oxidative stability is important. “ML2‐68,” “ML2‐72,” “US06,” “US1‐DL,” and “US1‐3” offer versatile options with their moderate oil content and fatty acid profiles that resist oxidation. This research highlights the potential for breeding sesame mutants tailored to specific applications, contributing to both enhanced oil production and improved nutritional value. The insights gained here pave the way for the development of sesame varieties with superior nutritional and functional attributes, thereby promoting the sustainable production and consumption of sesame oil.

Evaluating the effects of virulence genotype, swarming motility, and multi-locus sequence types of Escherichia coli on layer chicken embryos.

To determine the effects of swarming motility (SM) and multi-locus sequence types (MLST) on the main effect of virulence genotype of Escherichia coli through an embryos lethality assay between the 12th and 18th days of incubation. We collected 58 E. coli isolates from asymptomatic commercial hens (n=42) and lesions of colibacillosis cases (n=16), then classified their virulence genotype as avirulent, moderately virulent, virulent-healthy, and virulent-colibacillosis categories by the presence of five virulence-associated genes (iroN, ompT, hlyF, iutA, and iss). These isolates were further classified as non-motile, motile, or hyper-motile by SM assay. From the 58 isolates, we selected 29 for ELA and determined their MLST. Each isolate was inoculated into 15 embryonated eggs through the allantoic cavity. We found the avirulent isolates reduced the relative embryo weight compared to virulent-colibacillosis and moderately virulent isolates (37.49vs. 41.51 and 40.34%, P=0.03). Among the moderately virulent and virulent-colibacillosis categories, embryo lethality was lower when isolates were non-motile. Yolk retention was unaffected by virulence categories, motility, or MLST. Interaction between virulence genotype and SM substantially influenced the embryo lethality assay of E. coli isolates.

Genotype-by-environment interaction and stability analysis of grain yield of bread wheat (Triticum aestivum L.) genotypes using AMMI and GGE biplot analyses

Bread wheat is a vital staple crop worldwide; including in Ethiopia, but its production is prone to various environmental constraints and yield reduction associated with adaptation. To identify adaptable genotypes, a total of 12 bread wheat genotypes (G1 to G12) were evaluated for their genotype-environment interaction (GEI) and stability across three different environments for two years using Additive Main Effect and Multiplicative Interaction (AMMI) and genotype main effect plus genotype-by-environment interaction (GGE) biplots analysis. GEI is a common phenomenon in crop improvement and is of significant importance in genotype assessment and recommendation. According to combined analysis of variance, grain yield was considerably impacted by environments, genotypes, and GEI. AMMI and GGE biplots analysis also provided insights into the performance and stability of the genotypes across diverse environmental conditions. Among the 12 genotypes, G6 was selected by AMMI biplot analysis as adaptive and high-yielding genotype; G5 and G7 demonstrated high stability and minimal interaction with the environment, as evidenced by their IPCA1 values. G7 was identified as the most stable and high-yielding genotype. The GGE biplot's polygon view revealed that the highest grain yield was obtained from G6 in environment three (E3). E3 was selected as the ideal environment by the GGE biplot. The top three stable genotypes identified by AMMI stability value (ASV) were G5, G7, and G10, while the most stable genotype determined by Genotype Selection Index (GSI) was G7. Even though G6 was a high yielder, it was found to be unstable according to ASV and ranked third in stability according to GSI. Based on the study's findings, the GGE biplot genotype view for grain yield identified Tay genotype (G6) to be the most ideal genotype due to its high grain yield and stability in diverse environments. G7 showed similar characteristics and was also stable. These findings provide valuable insights to breeders and researchers for selecting high-yielding and stable, as well as high-yielding specifically adapted genotypes.

GGE analysis and stability of traits in tomato cultivars grown under organic farming conditions: A two-year study

The increasing conversion of agricultural land to organic farming requires the development of specifically adapted cultivars. So far, in tomato there is lack of research for selection of germplasm suitable for sustainable agroecosystems. In this study, we investigated the genotypic and environmental factors affecting the variation of plant, fruits, and root traits in 39 tomato genotypes grown under organic farming conditions. Four independent experiments were conducted in Italy and Spain across two consecutive seasons in 2019 and 2020. For all traits, the factorial linear regression model to estimate the main effects of genotype (G), location (L), year of cultivation (Y) and their interactions, revealed highly significant (P < 0.001) variations, with the G factor being largely predominant for most traits. The implementation of the “which-won-where”, “mean performance versus stability” and “discriminative vs representativeness” patterns in the GGE (Genotype plus Genotype by Environment interaction) analysis, allowed the identification of superior cultivars with high stability across the testing environments. Genomic characterization with 30 890 high quality SNPs from ddRADseq genotyping analysis, revealed that a specific cluster of cherry tomato accessions were low performing in terms of yield and fruit weight, on the contrary, showed a high content of soluble solids, which in agreement with GGE analysis. Results of this study provide a framework for the potential use of this locally adapted tomato germplasm to address the needs of more sustainable agriculture.

Investigating the dynamics of upland rice (Oryza sativa L.) in rainfed agroecosystems: an in-depth analysis of yield gap and strategic exploration for enhanced production

IntroductionAddressing the global demand for rice production necessitates innovative approaches to enhance upland rice yield in rainfed agroecosystems, considering the challenges posed by increasing population, limited land fertility, low productivity, and water availability.MethodsIn this study, our study investigated the impact of biochar and organic fertilizer on ten promising rice lines (G1 – G10) and two control (G11 – G12) cultivars under rainfed conditions. The experimental design used a split-plot design with four soil amendments as main plots, namely control, organic fertilizer, biochar, and biochar + organic fertilizer and 12 rice genotypes as subplot.ResultsThe absolute attainable yield gaps, differentiating organic and control (GAP1), biochar + organic and control (GAP2), and biochar and control (GAP3), ranged from 1.5 to 3.7 or increased of 91–580%, 0.8 to 3.5 (72–560%), and 0.6 to 2.58 tons/ha (58–472%), respectively. Notably, G2 + organic exhibited the highest positive absolute yield gap, ranging from 1.1 to 5.38 tons/ha, based on the yield gap matrix. Furthermore, genotype main effect plus genotype-environment interaction (GGE) biplot analysis identified G2 as the most promising rice line, displaying superior yield performance for cultivation in biochar and organic amended soils.DiscussionThese findings provide valuable insights for farmers, governments, and stakeholders, offering a roadmap to optimize rainfed areas for rice production, serving as practical guidance to enhance overall rice productivity in rainfed agroecosystems.

Envirotype-based delineation of environmental effects and genotype × environment interactions in Indian soybean (Glycine max, L.)

Soybean is a rainfed crop grown across a wide range of environments in India. Its grain yield is a complex trait governed by many minor genes and influenced by environmental effects and genotype × environment interactions. In the current investigation, grain yield data of different sets of 41, 30 and 48 soybean genotypes evaluated during 2019, 2020 and 2021, respectively across 19 locations and twenty years’ data on 19 different climatic parameters at these locations was used to study the environmental effects on grain yield, to understand the genotype × environment interactions and to identify the mega-environments. Through analysis of variance (ANOVA), it was found that predominant portion of the variation was explained by environmental effects (E) (53.89, 54.86 and 60.56% during 2019, 2020 and 2021, respectively), followed by genotype × environment interactions (GEI) (31.29, 33.72 and 28.82% during 2019, 2020 and 2021, respectively). Principal Component Analysis (PCA) revealed that grain yield was positively associated with RH (Relative humidity at 2 m height), FRUE (Effect of temperature on radiation use efficiency), WSM (Wind speed at 2 m height) and RTA (Global solar radiation based on latitude and Julian day) and negatively associated with VPD (Deficit of vapour pressure), Trange (Daily temperature range), ETP (Evapotranspiration), SW (Insolation incident on a horizontal surface), n (Actual duration of sunshine) and N (Daylight hours). Identification of mega-environments is critical in enhancing the selection gain, productivity and varietal recommendation. Through envirotyping and genotype main effect plus genotype by environment interaction (GGE) biplot methods, nineteen locations across India were grouped into four mega-environments (MEs). ME1 included five locations viz., Bengaluru, Pune, Dharwad, Kasbe Digraj and Umiam. Eight locations—Anand, Amreli, Lokbharti, Bidar, Parbhani, Ranchi, Bhawanipatna and Raipur were included in ME2. Kota and Morena constitutes ME3, while Palampur, Imphal, Mojhera and Almora were included in ME4. Locations Imphal, Bidar and Raipur were found to be both discriminative and representative; these test locations can be utilized in developing wider adaptable soybean cultivars. Pune and Amreli were found to be high-yielding locations and can be used in large scale breeder seed production.

Hydroponic Screening at Early Seedling Stage Identified Sources of Salinity Tolerance in Wheat (Triticum aestivum L.) Crop

Wheat is a vital crop globally, essential for agriculture, economics, and food security. However, in arid and semi-arid conditions, wheat production faces significant challenges due to low water availability, uneven rainfall distribution, and high soil salinity. The germination and early seedling stages are particularly vulnerable to these stresses. Therefore, this study assessed 15 wheat genotypes for their tolerance to salinity stress during early growth stages, using a hydroponic system with four salt stress levels (0, 50, 100, and 150 mM NaCl). Significant differences were observed for genotype and salinity main effects and their interaction on all investigated traits, indicating considerable variability in the response to salt stress among the investigated wheat cultivars. High NaCl concentrations led to substantial reductions in measured parameters across genotypes, with some showing resilience while others exhibited heightened sensitivity. Stress tolerance indices, such as mean productivity (MP), geometric mean productivity (GMP), harmonic mean (HM), stress tolerance index (STI) and yield index (YI), were identified as reliable indicators for selecting salt-tolerant wheat cultivars. Consequently, Sidi Okba (G11), Ziad (G12), Tamezghida (G13) and Zidane (G14) emerged as the most promising, displaying acceptable performance under both non-stress and salt-stress conditions. These genotypes could serve as valuable genetic resources for breeding programs aimed at enhancing wheat’s salinity tolerance, particularly in arid and semi-arid regions.

Sex-Dependent Regulation of Mucin Gene Transcription and Goblet Cell Secretion Machinery Following Intra-Airway IL-13 in Mice with Conditional Loss of Club Cell Creb1

IL-13 is an important effector molecule in allergic asthma, converting secretoglobin-positive club cells into mucin-secreting goblet cells and promoting mucin hypersecretion. IL-13-mediated conversion of club cells to goblet cells requires decreased gene expression of forkhead box A2 ( FOXA2) and increased gene expression of SAM pointed domain containing ETS transcription factor ( SPDEF). In addition, IL-13-mediated mucin hypersecretion may include modulation of purinergic and muscarinic receptors that control basal and stimulated mucus secretion, respectively. We recently found that the cAMP response element-binding protein (CREB), a ubiquitous transcription factor, directly binds to FOXA2 and modulates mucus secretion mechanisms in mice. Here we tested the hypothesis that conditional loss of club cell Creb1 in murine airways modulates the pro-mucin effects of IL-13. Loss of club cell Creb1 augmented IL-13-mediated increases in gene expression of the major gel-forming secreted mucins Muc5ac and Muc5b in male murine airways, with no effect on IL-13-mediated increases in Muc5ac and Muc5b in female murine airways. Examination of mucin secretion mechanisms revealed that IL-13 decreased muscarinic 3 receptor ( M3R) mRNA expression in male murine airways, whereas loss of club cell Creb1 prevented this, resulting in a significant treatment by genotype interaction. Interestingly, in female airways, a main effect of genotype was observed, with loss of club cell Creb1 reducing M3R mRNA expression in the airway. Examination of the purinergic receptor P2Y ( P2ry2) mRNA revealed a main treatment effect for IL-13 to increase P2ry2 expression in male murine airways with no impact of loss of club cell Creb1. In the females, IL-13 increased P2ry2 mRNA in both genotypes, with loss of club cell Creb1 significantly blunting the effect of IL-13 on P2ry2 mRNA. Lastly, we examined goblet cell density and mucin secretion mechanisms using Alcian Blue-PAS staining post in vivo stimulation with a muscarinic agent. We found no major effect of treatment or genotype in the density of goblet cells containing acidic mucins in either male or female airways. In contrast, both males and females showed IL-13-mediated increases in the density of goblet cells containing neutral mucins that was not impacted by genotype. Our preliminary findings suggest that loss of club cell Creb1 decreases molecular mucin secretion mechanisms in male and female airways, and increases Muc5ac and Muc5b mRNA in male airways, though a corresponding genotype-dependent increase in goblet cell density post cholinergic stimulation was not observed. Thus, we conclude that loss of club cell Creb1 causes changes to molecules important for IL-13-mediated mucin hypersecretion, without apparent functional benefit or detriment to the murine airway. This work was supported by the National Institutes of Health (OD026582, HL152101), the Cystic Fibrosis Foundation (REZNIKO20I0, REZNIKO19I0), the Katie Rose Foundation (AWD08203), and the University of Florida McKnight Career Accelerator Award. Angelina Bonilla was funded by the National institutes of Health (R25GM115298). Pedro Trevizan Bau receives the University of Florida McKnight Brain Institute Gator NeuroScholar Fellowship. This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

Breath-by-breath Analysis of a Rat Model of Pompe Disease Exposes Deficits in High-frequency Behaviors

Pompe disease (PD) is caused by a loss of function of the enzyme acid-α-glucosidase (GAA) leading to glycogen accumulation, neuromuscular dysfunction, and breathing failure. Here, we characterized longitudinal changes in breathing of GAA null ( Gaa−/−) rats on a breath-by-breath basis using a novel respiratory event detection algorithm. Adult Pompe (n = 5) and Sprague Dawley (n = 5) rats were implanted with chronic diaphragm EMG electrodes. Once a month from 4-10 months of age, full body plethysmography with concurrent EMG was used to record respiratory waveforms under room air and hypoxia. Waveforms were analyzed by the Adjustable Baselines Respiratory Analysis Program (ABRAP) algorithm. An adaptive threshold identified “respiratory events” defined as crossings of a threshold set between the recent maxima and minima of the waveform. After event identification, a range of waveform characteristics are calculated and tagged to that event. Averages of each characteristic were taken for all events and then analyzed based on respiratory rate in breaths/minute (bpm): low (&lt;120 bpm, ‘quiet breathing’), medium (between 120 and 240 bpm) and high (&gt;240 bpm, ‘high frequency sniffng’). In room air, Pompe rats had a lower breathing rate than wildtype rats by month 10 (173 vs 285 bpm, standard error of the difference (SE of diff.) 37, p=0.03). This decrease in frequency was driven by the high rate events as there were no differences in the mean rate of low rate events at month 10 (80 vs 83 bpm, SE of diff. 3) while the frequency of high rate events was significantly lower by 9 months (mo 9: 393 vs 440 bpm, SE of diff. 16, p=0.04, mo 10: 383 vs 440 bpm, SE of diff. 17, p=0.008). Not only was the breathing frequency of high-rate events impaired in Pompe rats, but there was a trend towards decrease in the proportion of events falling in the high rate category indicating less time spent in high rate breathing behaviors (mo 4: 52 vs 59%, SE of diff. 6, mo 10: 25 vs 52%, SE of diff. 9) with a concomitant increase in the proportion of low frequency events. Additionally, there was a main effect of genotype on the latency between diaphragm activation and onset of inspiratory flow during hypoxia with Pompe rats having a longer latency across all months and frequency bands. This difference was most pronounced in the medium (44 vs 29 ms, SE of diff. 4 ms, p=0.003) frequency events. Active expiration was impaired in Pompe animals with peak post-event airflow (maximum positive pressure reached before the next inspiratory effort) blunted across all months. This effect was pronounced in the high frequency events (0.16 mL vs 0.24 mL, SE of diff. 0.01, p=0.0001). These deficits in high frequency events are consistent with progressive neuromuscular diaphragm weakness. We conclude that comprehensive analyses of all respiratory events over extended recording periods enables detection of altered breathing behaviors that may be missed when assessing only short periods of quiet breathing. In turn, this may help uncover a respiratory signature of disease progression in Pompe disease or other neuromuscular conditions. 2R01HD052682-11A1 (DDF, BJB), T32HL134621 (AM), R01HL153102 (ED), SPARC OT2OD023854 (ED), Craig H. Neilsen Pilot Grant (ED). This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

Trend in yield and protein content relationship in a historical series of durum wheat varieties under rainfed and irrigated environments

Improving grain yield (GY) is a major goal of durum wheat breeding programs, and a high yield needs to be attained while preserving grain quality. The current study evaluated the changes caused by a breeding program on the association between GY and grain protein content (PC) in Moroccan durum wheat. Field experiments with six leading varieties released over three decades were conducted for three growing seasons under two water regimes. GY and PC were largely controlled by environmental effects, including both the growing season and water regime. The year of release and growing season effects were the predominant sources of variation in total chlorophyll content (TCC). A significant increase in GY with the year of release was observed under irrigated conditions, whereas a significant increase in PC over time was noted under rainfed conditions. In addition, a negative correlation was revealed between GY and PC, which was significant only in the rainfed trials. Similarly, a significant and positive association was noted between PC and TCC within the rainfed trials, but not in the irrigated trials. Based on GGE (genotype main effect plus genotype ​× ​environment interaction) biplot analysis, the new varieties ‘Marouane’ and ‘Faraj’ were identified as desirable varieties in terms of GY and PC close to the ‘ideal genotype’. These results indicated that when grown under current irrigated conditions, new varieties improved by the Moroccan breeding program showed better GY performance than old varieties.

Adrenal circadian clock protein BMAL1 regulates aldosterone and renal excretory function in a sex- and time-dependent manner

BMAL1 is a core circadian clock protein that is important for circadian rhythms of physiological function, as well as maintaining physiological homeostasis, including aspects of kidney function. Global Bmal1 knockout (KO) male mice exhibit loss of diurnal rhythm of renal sodium (Na) excretion in response to a normal or high salt diet. However, renal Na excretion rhythm remained intact in kidney-specific Bmal1 KO male mice. The adrenal gland synthesizes hormones, such as aldosterone, which influence renal electrolyte handling. The role of adrenal BMAL1 in the regulation of renal function remains unknown. The study objective was to test the hypothesis that adrenal BMAL1 is required for normal rhythms of aldosterone and renal Na handling. Male and female adrenal-specific aldosterone synthase Cre positive (AS) Bmal1 KO and littermate Cre negative, floxed control mice ( n=5-9/genotype) were acclimated to metabolic cages for 3 days then samples were collected for three days with mice on a normal salt diet followed by 7 days on a low salt diet to challenge the mice to stimulate aldosterone production. Twelve-hour food intake was measured, and urine samples were collected to measure night/day patterns in renal Na balance. Urinary aldosterone excretion was measured by ELISA. Both day and night urinary aldosterone excretion were increased following a low salt diet in control and KO male mice (ANOVA main effect of diet, p&lt;0.05). Male AS Bmal1 KO mice displayed a significant increase in day urinary aldosterone excretion following a normal and low salt diet (ANOVA main effect of genotype, p=0.0230), with a trend for an increase in night urinary aldosterone excretion ( p=0.0571). Interestingly, these genotype effects were not seen in female AS Bmal1 KO mice. Urinary aldosterone was increased following a low salt diet in female mice, but only during the nighttime in control and KO mice ( p=0.0205). Sex differences were also observed in renal Na balance, where Na balance was more positive during the day in male, but not female, AS Bmal1 KO vs. control mice (ANOVA main effect of genotype, p=0.0282). Together, these data suggest adrenal BMAL1 is required for normal rhythms of aldosterone and renal Na balance in a sex-dependent manner. Future work will investigate whether the changes in aldosterone contribute to changes in renal Na balance. This work was supported by the American Heart Association (Postdoctoral FellowshipP0240441 to H.C.; Established Investigator Award 19EIA34660135 to M.G), theRobert and Mary Cade Professorship in Physiology (to M.G.) and the NIH(F32DK121424 to G.R.C.). This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

Differences in colostrum components of Katahdin ewes varies with post-weaning FEC EBV

Variation in post-weaning fecal egg count (PFEC) estimated breeding values (EBV) has revealed effects beyond the reduction of parasitism. Previous work demonstrated lambs with a PFEC of < −50 (Low-PFEC) have higher survivability to weaning than lambs with a PFEC of > +50 (High-PFEC). In response to clostridial vaccination, Low-PFEC lambs also generate a more robust and rapid antibody response. These findings prompted curiosity of the maternal influence underpinning differences observed in lamb survivability based upon PFEC genotype. Thus, this study aimed to assess the impact of dam PFEC EBV on IgG concentration, Brix values, and color within colostrum. For Year 1 (YR1), Katahdin ewes from the Virginia Tech SWAREC were selected based on the following criteria: age (3–6 years), have a Katahdin Hair Index EBV (>104, pre-genomic EBV), and be twin bearing. Ewes fitting those criteria were sorted by their PFEC EBV into two groups: Low-PFEC (PFEC < −50; n=20) and High-PFEC (PFEC > +50; n=18) and sampled at three distinct time points, early (within 24-hr post-partition), mid (30-d post-parturition) and late (60-d post-parturition) lactation. For Year 2 (YR2), all lactating ewes at the SWAREC were analyzed and were sorted into three groups: Low-PFEC (PFEC < −50; n=39), Mid-PFEC ( −49 < PFEC < +49; n=38), and High-PFEC (PFEC > +50; n=23) and colostrum samples were taken within 6-hrs post-partition. All colostrum and milk samples were collected by milking 10 mL from each half of the ewes' udder. Total IgG within colostrum and milk was measured using an ovine-specific enzyme-linked immunosorbent assay (ELISA). In YR1, main effects of ewe genotype revealed that Low-PFEC ewes had a greater concentration of IgG across all lactation time points (631 ng/mL vs 222 ng/mL; P < 0.0001). This was primarily driven by IgG differences within colostrum, where a 2.5 fold difference in IgG were observed between Low-PFEC and High-PFEC groups (937 ng/mL vs. 319 ng/mL, P < 0.0001). In YR2, a stepwise decrease in average colostrum IgG concentration from Low-PFEC to High-PFEC groups was observed (825 ng/mL, 513 ng/mL, and 258 ng/mL, respectively; P < 0.0429). Likewise, a stepwise decrease in yellow to blue hue or b* value occurs from Low-PFEC to High-PFEC groups (18.97, 16.91, and 15.22, respectively; P < 0.0459). No correlations were found between IgG and Brix values within the Low-PFEC and High-PFEC grouping; remarkably, Mid-PFEC ewes have a slightly positive correlation between IgG and Brix (r= 0.34, P = 0.0334). These data indicate that Low-PFEC Katahdin ewes generate colostrum with greater antibody concentration and more yellow color. Lambs born to these ewes would have a significant advantage in passive immunity from their dams, which may explain some component of the enhanced lamb survivability observed pre-weaning.

The emergence of inflammatory microglia during gut inflammation is not affected by FFAR2 expression in intestinal epithelial cells or peripheral myeloid cells

Gut inflammation can trigger neuroinflammation and is linked to mood disorders. Microbiota-derived short-chain fatty acids (SCFAs) can modulate microglia, yet the mechanism remains elusive. Since microglia do not express free-fatty acid receptor (FFAR)2, but intestinal epithelial cells (IEC) and peripheral myeloid cells do, we hypothesized that SCFA-mediated FFAR2 activation within the gut or peripheral myeloid cells may impact microglia inflammation. To test this hypothesis, we developed a tamoxifen-inducible conditional knockout mouse model targeting FFAR2 exclusively on IEC and induced intestinal inflammation with dextran sodium sulfate (DSS), a well-established colitis model. Given FFAR2′s high expression in myeloid cells, we also investigated its role by selectively deleting it in these populations of cells. In an initial study, male and female wild-type mice received 0 or 2% DSS for 5d and microglia were isolated 3d later to assess inflammatory status. DSS induced intestinal inflammation and upregulated inflammatory gene expression in microglia, indicating inflammatory signaling via the gut-brain axis. Despite the lack of significant effects of sex in the intestinal phenotype, male mice showed higher microglial inflammatory response than females. Subsequent studies using FFAR2 knockout models revealed that FFAR2 expression in IECs or immune myeloid cells did not affect DSS-induced colonic pathology (i.e. clinical and histological scores and colon length), or colonic expression of inflammatory genes. However, FFAR2 knockout led to an upregulation of several microglial inflammatory genes in control mice and downregulation in DSS-treated mice, suggesting that FFAR2 may constrain neuroinflammatory gene expression under healthy homeostatic conditions but may permit it during intestinal inflammation. No interactions with sex were observed, suggesting sex does not play a role on FFAR2 potential function in gut-brain communication in the context of colitis. To evaluate the role of FFAR2 activated by microbiota-derived SCFAs, we employed the same knockout and DSS models adding fermentable dietary fiber (0 or 2.5% inulin for 8 wks). Despite no genotype or fiber main effects, contrary to our hypothesis, inulin feeding augmented DSS-induced inflammation and signs of colitis, suggesting context-dependent effects of fiber. These findings highlight microglial involvement in colitis-associated neuroinflammation and advance our understanding of FFAR2′s role in the gut-brain axis. Although not integral, we observed that the role of FFAR2 differs between homeostatic and inflammatory conditions, underscoring the need to consider different inflammatory conditions and disease contexts when investigating the role of FFAR2 and SCFAs in the gut-brain axis.