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Product bundling is receiving increasing attention in sustainable agricultural development as a means to ensure access to and enhance the uptake of, agricultural technologies by smallholders. Yet, the how and why of bundling for smallholders are not well-understood. The current paper, therefore, brings bundling theory from the marketing literature to the smallholder context. We use a conjoint experiment, a proven marketing technique for designing new products, services and bundles, to design agricultural input and service bundles for soybean-producing smallholders in rural Ethiopia. The empirical findings from 252 smallholders suggest that product bundling enhances smallholders’ preferences and hence intentions to adopt technologies but that bundles must be designed carefully following a smallholder-centric approach. Drawing on our findings and the literature, we delineate the different steps that need to be taken to develop bundles for the successful uptake of new technologies by smallholders. [ABSTRACT FROM AUTHOR]

The objective of this study was to evaluate the GreenFeed (GF) and respiration chambers (RC) for daily and intraday measurements of the enteric gaseous exchange, as well as the metabolic heat production, lying behavior, and feed intake (FI) rate of dairy cows at these 2 respective housing conditions [tie-stall barn (TS) vs. RC] during the summer periods. Sixteen multiparous lactating dairy cows were recruited and arranged in a randomized complete block design with a baseline period established for each cow. Cows were given a basal diet (CON) for a baseline period of 7 d and were then fed a 3-nitrooxypropanol (3-NOP)-containing feed for the subsequent 26 d as experimental period. During both the baseline and the last 7 d of treatment period, gaseous exchanges of each animal were measured in the TS using GF for 8 6-hourly staggered measurements over 3 d, immediately followed by the measurement in RC for 2 d. Corresponding DMI, milk yield, and behavior parameters (e.g., lying behavior and FI rate) in TS and RC were recorded. The correlation coefficients of CH 4 and H 2 using raw data were 0.84 and 0.85, respectively. For all gases, correlation coefficients between GF and RC on individual cow level decreased when the marginal fixed effects (e.g., inhibitor and breed) were corrected by a mixed model. There were no differences in daily CH 4 production or intensity between GF and RC (442 vs. 443 g CH 4 /d or 16.6 vs. 16.2 g CH 4 /kg MY). However, greater CH 4 yield was measured by GF than RC (19.0 vs. 17.8 g CH 4 /kg DMI), driven by a lower DMI (23.3 vs. 24.6 kg/d) when cows were housed in TS sampled by GF compared with cows being housed and sampled in RC. The correlations for CO 2 production and O 2 consumption were moderate and expected due to the variation associated with the mild heat stress condition during GF measurements in the TS (Thermal humidity index (THI) 56 vs. 68), as indicated by the reduced lying time (-2.1 h/d). At the intraday level, there was an interaction between techniques and hour-of-day for CH 4 production, as indicated by the discrepancies in post-prandial CH 4 emissions between techniques. In summary, this set of results showed that there were strong positive correlations for CH 4 and H 2 emissions between GF and RC based on individual cow data. However, such relationship should be interpreted with caution, given the data clustering resulting from the use of inhibitor 3-NOP. On treatment level, these 2 techniques detected similar inhibitor effect on the estimated daily CH 4 emissions. The intraday patterns of CH 4 and H 2 production captured by GF provided a close approximation for those measured by RC. Nevertheless, potential underestimation may occur, especially following fresh feed delivery. For measuring CO 2 production and O 2 consumption, the GF captured similar intraday variations to those in the RC. However, the estimated daily production and consumption were not directly comparable, which was expected due to the variable thermal conditions during the summer. Further evaluations under the same weather conditions are warranted., (The Authors. Published by Elsevier Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)

Emulsifiers are essential for achieving a homogenous distribution of lipophilic supplements in in vitro rumen fluid incubations. Since emulsifiers can alter rumen fermentation, it is crucial to select one that minimally impacts fermentation parameters to reduce potential biases. This study aimed to evaluate seven emulsifiers' impact on in vitro ruminal fermentation using the Hohenheim Gas Test in order to identify the most inert emulsifier. Rumen fluids were collected from three non-lactating Original Brown-Swiss cannulated cows before morning feeding and incubated for 24 h with a basal diet in triplicates. The emulsifiers tested were ethanol, ethyl acetate, propylene glycol, glycerol, ethylene glycol, soy lecithin, and Tween® 80, each in two dosages (0.5% or 1% v/v). The untreated basal diet served as control. Compared to control, in vitro organic matter digestibility was enhanced by ethyl acetate (by 36.9 and 48.2%), ethylene glycol (by 20.6 and 20.1%), glycerol (by 46.9 and 56.8%) and soy lecithin (by 19.7 and 26.8%) at 0.5 and 1% dosage, respectively. Additionally, the 24-h methane production increased for ethanol (by 41.9 and 46.2%), ethylene glycol (by 50.5 and 51.5%), and glycerol (by 63.1 and 65.4%) for the 0.5 and 1% dosage, respectively, and 0.5% dosage for ethyl acetate (by 31.6%). The acetate molar proportion was 17.2%pt higher for ethyl acetate, and 25.5%pt lower for glycerol at 1% dosage, compared to the control. The propionate concentration was 22.1%pt higher 1% glycerol, and 15.2%pt and 15.1%pt higher for 0.5 and 1% propylene glycol, respectively, compared to the control. In summary, Tween® 80 did not significantly affect in vitro rumen fermentation parameters, making it the most suitable choice for in vitro incubations involving lipophilic substances in rumen fluid. Ethanol may be considered as an alternative emulsifier if methane production is not the variable of interest., (© 2024 The Authors. Journal of Animal Physiology and Animal Nutrition published by Wiley‐VCH GmbH.)

Background: This case details a 44-year-old man with end-stage ischemic cardiomyopathy with refractory ventricular tachycardia (VT). The patient has a single-chamber implantable cardioverter-defibrillator, has had 2 VT ablations, and uses medication to manage his VT. Despite these interventions, he continued to have episodes of VT. The patient underwent stereotactic body radiation therapy (SBRT) to help reduce the burden of his VT. The patient received a dose of 25 Gy to his right inferior lateral region of the heart and a dose of 15 Gy to the inferior portion of the heart closer to the stomach. The patient followed up 1 month later and reported that his energy levels improved and that no arrhythmias had occurred since his SBRT treatment., Discussion: The options for treating end-stage VT are limited. However, a treatment option using SBRT has been introduced to reduce the VT burden in patients. Cardiac SBRT is a noninvasive outpatient procedure that, while still awaiting U.S. Food and Drug Administration approval, reduces arrhythmia episodes and offers favorable short-term benefits for patients who have not responded to traditional treatment modalities., Conclusion: Cardiac SBRT is a novel treatment for VT in patients where standard treatments have failed. This case study demonstrates that SBRT effectively reduced arrhythmias in a patient with VT. The long-term clinical outcomes are not known, but the opportunity SBRT offers for treatment-refractory patients is favorable and should be considered., (© 2024 American Society of Radiologic Technologists.)

Alzheimer's disease (AD) is a progressive and incurable neurodegenerative disease characterized by the extracellular deposition of amyloid plaques. Investigation into the composition of these plaques revealed a high amount of amyloid-β (Aβ) fibrils and a high concentration of lipids, suggesting that fibril-lipid interactions may also be relevant for the pathogenesis of AD. Therefore, we grew Aβ40 fibrils in the presence of lipid vesicles and determined their structure by cryo-electron microscopy (cryo-EM) to high resolution. The fold of the major polymorph is similar to the structure of brain-seeded fibrils reported previously. The majority of the lipids are bound to the fibrils, as we show by cryo-EM and NMR spectroscopy. This apparent lipid extraction from vesicles observed here in vitro provides structural insights into potentially disease-relevant fibril-lipid interactions., (© 2024. The Author(s).)

Little is known about how maturation of Alzheimer's disease-related amyloid β (Aβ) fibrils alters their stability and potentially influences their spreading in the brain. Using high-pressure NMR, we show that progression from early to late Aβ40 aggregates enhances the kinetic stability, while ageing during weeks to months enhances their thermodynamic stability.

Small molecules that bind to oligomeric protein species such as membrane proteins and fibrils are of clinical interest for development of therapeutics and diagnostics. Definition of the binding site at atomic resolution via NMR is often challenging due to low binding stoichiometry of the small molecule. For fibrils and aggregation intermediates grown in the presence of lipids, we report atomic-resolution contacts to the small molecule at sub nm distance via solid-state NMR using dynamic nuclear polarization (DNP) and orthogonally labelled samples of the protein and the small molecule. We apply this approach to α-synuclein (αS) aggregates in complex with the small molecule anle138b, which is a clinical drug candidate for disease modifying therapy. The small central pyrazole moiety of anle138b is detected in close proximity to the protein backbone and differences in the contacts between fibrils and early intermediates are observed. For intermediate species, the 100 K condition for DNP helps to preserve the aggregation state, while for both fibrils and oligomers, the DNP enhancement is essential to obtain sufficient sensitivity., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper. AG, AL, SR and CG are inventors in two patents (USA Pat., US20110293520A1 and Pat. WO 2021/099518) related to the diaryl pyrazole compounds, in particular anle138b. AG and CG are shareholders and co-founders of MODAG GmbH. AG is a full-time employee, AL and SR are part-time employees of MODAG GmbH., (Copyright © 2023 The Author(s). Published by Elsevier Inc. All rights reserved.)

Banana weevil (Cosmopolites sordidus, Germar) is a major pest in East African Highland Banana. The influence of crop nutritional status on weevil damage is poorly understood. Nutrient availability affects the nutritional quality of plants for weevils and may affect weevil damage. Here, we evaluate the effect of insecticides alone and in combination with fertilisers (N, P, K and Si) on weevil damage using data from two experiments in central and southwest Uganda. In the first experiment, we varied chlorpyrifos and application rates of N, P and K. In the second experiment, we varied the application rates of K and Si. Treatment effects were analysed using generalised linear mixed models with a negative binomial distribution. In the first experiment, chlorpyrifos reduced and N increased weevil damage, while P and K had no significant effect. In the K or Si application rates reduced weevil damage compared with the control. We conclude that the combined application of chlorpyrifos with K and Si fertilisers can contribute to weevil damage control on sites with low nutrient availability and should form part of integrated weevil management in bananas. Future studies should assess how much reduction in insecticide use is possible in EAHB with judicious input rates., Competing Interests: The authors have declared that they have no competing interests with respect to this work., (Copyright: © 2023 Bukomeko et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Amyloid-beta (Aβ) deposition as senile plaques is a pathological hallmark of Alzheimer's disease (AD). AD is characterized by a large level of heterogeneity in amyloid pathology, whose molecular origin is poorly understood. Here, we employ NMR spectroscopy and MD simulation at ambient and high pressures and investigate how AD-related mutations in Aβ peptide influence the stability of Aβ aggregates. The pressure-induced monomer dissociation from Aβ aggregates monitored by NMR demonstrated that the Iowa (D23N), Arctic (E22G), and Osaka (ΔE22) mutations altered the pressure stability of Aβ40 aggregates in distinct manners. While the NMR data of monomeric Aβ40 showed only small localized effects of mutations, the MD simulation of mutated Aβ fibrils revealed their distinct susceptibility to elevated pressure. Our data propose a structural basis for the distinct stability of various Aβ fibrils and highlights "stability" as a molecular property potentially contributing to the large heterogeneity of amyloid pathology in AD.

In the past decade, there has been fast-growing interest among researchers to discover bioactive peptides from edible insects and to evaluate their potential applications in the management of human, livestock, and plant health. This review summarizes current knowledge of insect-derived peptides and their potential role in tackling human health issues and solving agriculture problems by protecting crops and livestock against their pathogens. Numerous bioactive peptides have been identified from edible insect species, including peptides that were enzymatically liberated from insect proteins and endogenous peptides that occur naturally in insects. The peptides exhibited diverse bioactivities, encompassing antioxidant, anti-angiotensin-converting enzyme, anti-dipeptidyl peptidase-IV, anti-glucosidase, anti-lipase, anti-lipoxygenase, anti-cyclooxygenase, anti-obesity, and hepatoprotective activities. Such findings point to their potential contribution to solving human health problems related to inflammation, free radical damage, diabetes, hypertension, and liver damage, among others. Although most of the experiments were performed in vitro, evidence for the in vivo efficacy of some peptides is emerging. Evidence of the protective effects of insect-derived endogenous antimicrobial peptides in combating farm animal and plant pathogens is available. The ability of insect-derived endogenous neuropeptides to protect plants against herbivorous insects has been demonstrated as well. Nevertheless, the potency of peptides identified from insect protein hydrolysates in modulating livestock and plant health remains a knowledge gap to be filled.

Pomaces are food industry by-products and may serve as animal feed to increase sustainability of meat and milk production. The aim of the present study was to evaluate fermentation characteristics of dried fruit and vegetable pomaces in a short-term in vitro experiment using the Hohenheim Gas Test. A selection of six fruit (apple, aronia, orange, pomegranate, red, white grape) and three vegetable (beetroot, carrot, tomato) pomaces was tested in three concentrations (150, 300, 500 g kg -1 of dry matter (DM)) as supplement to the basal diet (hay, used as control). Three runs were performed, using rumen fluid from one of three different rumen-cannulated cows in each run. Per run, each compound was tested in duplicate. After 24 h incubation, total gas production, methane and CO 2 concentration, short-chain fatty acids, in vitro organic matter digestibility as well as microbial counts were determined. In addition, the pomaces' polyphenol content including the fractions non-tannin phenols, condensed tannins and hydrolysable tannins were analysed. Most pomaces did not significantly affect rumen fermentation characteristics in any of the tested dosages and may thus be applied in ruminant nutrition without adverse effects. Aronia significantly decreased (-14.5%) the organic matter digestibility in the highest concentration whereas apple (+12%), carrot (+10%) and beetroot (+8%) increased gas formation related to digestible organic matter. The 500 g kg -1 dosage of pomegranate significantly decreased methane formation by about 28% without impairing digestibility. Pomegranate was the only pomace of those high in total tannins that contained exceptionally high amounts of hydrolysable (90% of total tannins) and proportionally low amounts of condensed tannins (10% of total tannins), indicating that the hydrolysable tannins most likely reduced the methane production. Therefore, pomegranate pomace may be an interesting option for a methane mitigating feed supplement in ruminants and should be considered for following in vivo testing., (wileyonlinelibrary.com/journal/jpn.)

Excessive dietary intake of fats and sugars ("Western diet", WD) is one of the leading causes of obesity. The consumption of the microalga Arthrospira platensis (spirulina, Sp) is increasing due to its presumed health benefits. Both WD and Sp are also consumed by pregnant and breastfeeding women. This study investigated if gestating and lactating domestic pigs are an appropriate model for WD-induced metabolic disturbances similar to those observed in humans and if Sp supplementation may attenuate any of these adverse effects. Pigs were fed a WD high in fat, sugars, and cholesterol or a control diet. Half of the animals per diet group were supplemented with 20 g Sp per day. The WD did not increase body weight or adipose tissue accumulation but led to metabolic impairments such as higher cholesterol concentration in plasma, lower IGF1 plasma levels, and signs of hepatic damage compared to the control group. Spirulina supplementation could not reduce all the metabolic impairments observed in WD-fed animals. These findings indicate limited suitability of gestating and lactating domestic pigs as a model for WD but a certain potential of low-dose Sp supplementation to partially attenuate negative WD effects.

Scientific evidence suggests that not only murine scent communication is regulated by major urinary proteins, but that their expression may also vary in response to metabolism via a yet unknown mechanism. Major urinary proteins are expressed mainly in the liver, showing a sexually dimorphic pattern with substantially higher expression in males. Here, we investigate the metabolic implications of a major urinary protein knockout in twelve-week-old male and female C57BL/6N mice during ad libitum feeding. Despite both sexes of major urinary protein knockout mice displayed numerically increased body weight and visceral adipose tissue proportions compared to sex-matched wildtype mice, the main genotype-specific metabolic differences were observed exclusively in males. Male major urinary protein knockout mice exhibited plasma and hepatic lipid accumulation accompanied by a hepatic transcriptome indicating an activation of lipogenesis. These findings match the higher major urinary protein expression in male compared to female wildtype mice, suggesting a more distinct reduction in energy requirements in male compared to female major urinary protein knockout mice. The observed sex-specific anabolic phenotype confirms a role of major urinary protein in metabolism and, since major urinary proteins are not expressed in humans, suggests the major urinary protein knockout mouse as a potential alternative model for translational metabolism research which needs to be further elucidated., (© 2022. The Author(s).)

Partner recognition in protein binding is critical for all biological functions, and yet, delineating its mechanism is challenging, especially when recognition happens within microseconds. We present a theoretical and experimental framework based on straight-forward nuclear magnetic resonance relaxation dispersion measurements to investigate protein binding mechanisms on sub-millisecond timescales, which are beyond the reach of standard rapid-mixing experiments. This framework predicts that conformational selection prevails on ubiquitin's paradigmatic interaction with an SH3 (Src-homology 3) domain. By contrast, the SH3 domain recognizes ubiquitin in a two-state binding process. Subsequent molecular dynamics simulations and Markov state modeling reveal that the ubiquitin conformation selected for binding exhibits a characteristically extended C-terminus. Our framework is robust and expandable for implementation in other binding scenarios with the potential to show that conformational selection might be the design principle of the hubs in protein interaction networks., (© 2022. The Author(s).)

A solid understanding of the dynamics of plant nutrient requirements and uptake from the soil is needed to provide robust fertilizer recommendations, timing of applications and nutrient use efficiency. Our objective was to develop and test the ability of the crop model LINTUL-Cassava-NPK to simulate biomass growth and yield of cassava under nutrient-limited conditions. We used experimental data from six fields located in three different agro-ecologies in Nigeria: Rainforest Zone- Ogoja and Ikom (Cross River), Rainforest Transition Zone - Ekpoma (Edo) and Guinea Savanna Zone - Otukpo (Benue) over two consecutive growing seasons from 2016 to 2018. Nutrient stress in the model was implemented by combining N, P and K nutrition indices (NI) to account for the interaction of multiple nutrient limitations for crop growth. Nutrient uptake was determined by balancing demand and supply of nutrient equivalents. We parameterized and calibrated the model using observations from an experiment conducted under optimal growing conditions in Edo during the 2016 planting season. The model was then tested with data from experiments conducted in the 2017 season in Edo, Cross River and Benue. The model captured the uptake patterns of N, P and K well. Uptakes of N, P and K, and storage root yield were predicted with a small root mean squared error of 5.1 g N m -2 , 0.8 g P m -2 , 3.3 g K m -2 and 308 g DM roots m -2 , with an R of 0.7 - 0.8 for linear relationships between simulated and observed values. The time course of development of nutrient-limited yield of green leaves, stems and storage roots were simulated reasonably well. In general, the model responded aptly to both nutrient omissions and varying amounts of NPK. These findings increase our understanding of nutrient limitations and N, P and K interactions on cassava growth and yield. The model provided insight into surplus amounts of nutrients in the soil at the end of the season and, specifically, the need to balance the supply of N and K for cassava. To our knowledge, this is the first tested cassava process-based model that includes the three macro-nutrients.2 of 0.7 - 0.8 for linear relationships between simulated and observed values. The time course of development of nutrient-limited yield of green leaves, stems and storage roots were simulated reasonably well. In general, the model responded aptly to both nutrient omissions and varying amounts of NPK. These findings increase our understanding of nutrient limitations and N, P and K interactions on cassava growth and yield. The model provided insight into surplus amounts of nutrients in the soil at the end of the season and, specifically, the need to balance the supply of N and K for cassava. To our knowledge, this is the first tested cassava process-based model that includes the three macro-nutrients., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2022 The Authors.)

Feeding high-energy (HED) or high-fat diets during gestation and lactation to pigs may help cover the energy requirements of high-prolific sows but may also adversely affect their reproductive performance. The microalga Arthrospira plantensis (Sp), rich in bioactive compounds, has been described to exert beneficial health effects. The present study investigated the effects of HED and Sp intake during gestation and lactation in pigs. Twenty-four primiparous crossbred sows were fed either a HED or a control diet. Half of the sows per group were supplemented with 20 g/day of Sp. Despite a higher gross energy intake, consuming the HED did not affect the sows' reproductive and lactational performance but significantly modified the colostrum fatty acid (FA) composition and tended to decrease the number of weaned piglets. The Sp supplementation did not affect the reproduction and lactation traits, but slightly affected the colostrum FA composition. A trend was observed for an interaction of diet and Sp in terms of offspring sex ratio with a 50% lower male-to-female ratio in the HED group compared to all other animals. These findings suggest that an HED and Sp intake hardly influence reproduction in sows. However, the HED modified the colostrum FA composition, whereas the Sp had only fewer effects, which may potentially affect offspring performance.

Alzheimer's disease is characterized by the presence of extraneuronal amyloid plaques composed of amyloid-beta (Aβ) fibrillar aggregates in the brains of patients. In mouse models, it has previously been shown that atorvastatin (Ator), a cholesterol-lowering drug, has some reducing effect on the production of cerebral Aβ. A meta-analysis on humans showed moderate effects in the short term but no improvement in the Alzheimer's Disease Assessment Scale-Cognitive Subscale behavioral test. Here, we explore a potential direct effect of Ator on Aβ42 aggregation. Using NMR-based monomer consumption assays and CD spectroscopy, we observed a promoting effect of Ator in its original form (Ator-calcium) on Aβ42 aggregation, as expected because of the presence of calcium ions. The effect was reversed when applying a CaCO 3 -based calcium ion scavenging method, which was validated by the aforementioned methods as well as thioflavin-T fluorescence assays and transmission electron microscopy. We found that the aggregation was inhibited significantly when the concentration of calcium-free Ator exceeded that of Aβ by at least a factor of 2. The 1 H- 15 N heteronuclear single quantum correlation and saturation-transfer difference NMR data suggest that calcium-free Ator exerts its effect through interaction with the 16 KLVF 19 binding site on the Aβ peptide via its aromatic rings as well as hydroxyl and methyl groups. On the other hand, molecular dynamics simulations confirmed that the increasing concentration of Ator is necessary for the inhibition of the conformational transition of Aβ from an α-helix-dominant to a β-sheet-dominant structure., Competing Interests: Conflict of interest The authors declare that they have no conflicts of interest with the contents of this article., (Copyright © 2022 The Authors. Published by Elsevier Inc. All rights reserved.)

The voltage-dependent anion channel (VDAC), the most abundant protein in the outer mitochondrial membrane, is responsible for the transport of all ions and metabolites into and out of mitochondria. Larger than any of the β-barrel structures determined to date by magic-angle spinning (MAS) NMR, but smaller than the size limit of cryo-electron microscopy (cryo-EM), VDAC1's 31 kDa size has long been a bottleneck in determining its structure in a near-native lipid bilayer environment. Using a single two-dimensional (2D) crystalline sample of human VDAC1 in lipids, we applied proton-detected fast magic-angle spinning NMR spectroscopy to determine the arrangement of β strands. Combining these data with long-range restraints from a spin-labeled sample, chemical shift-based secondary structure prediction, and previous MAS NMR and atomic force microscopy (AFM) data, we determined the channel's structure at a 2.2 Å root-mean-square deviation (RMSD). The structure, a 19-stranded β-barrel, with an N-terminal α-helix in the pore is in agreement with previous data in detergent, which was questioned due to the potential for the detergent to perturb the protein's functional structure. Using a quintuple mutant implementing the channel's closed state, we found that dynamics are a key element in the protein's gating behavior, as channel closure leads to the destabilization of not only the C-terminal barrel residues but also the α2 helix. We showed that cholesterol, previously shown to reduce the frequency of channel closure, stabilizes the barrel relative to the N-terminal helix. Furthermore, we observed channel closure through steric blockage by a drug shown to selectively bind to the channel, the Bcl2-antisense oligonucleotide G3139.

Female mammalian reproductive functions are closely linked to body condition and metabolic status. Energy homeostasis is regulated by endocrine hormones such as insulin, IGF-I, leptin, and adiponectin via the hypothalamic-pituitary-adrenal axis. These metabolic hormones and their receptors are also expressed in reproductive tissues and the embryo. We investigated the relationship between circulating leptin and the fatty acid (FA) and amino acid (AA) composition of the equine uterine fluid (UF) and peripheral blood plasma (BP) by using a mass spectrometry-based approach. UF and BP were collected from ten broodmares on days 6 and 7 post ovulation, respectively. The mares were retrospectively assigned to two groups according to their BP leptin concentrations (high leptin [> 1.6 ng/mL] versus low leptin [<0.8 ng/mL]). Specific AA and FA compositions for BP and UF were found with different levels of respective metabolite abundances. The main FAs in BP were stearic, palmitic and linoleic acid. In UF, the three most abundant FAs were eicosapentaenoic, arachidonic and stearic acid. The AA profile of BP was dominated by glycine, glutamine, serine and alanine, which were likewise among the highly abundant AAs in UF. In UF, glutamic acid had by far the highest concentration. Therefore, BP leptin concentration within a physiological range does not seem to affect the specific FA nor the AA composition of the UF. The composition of the UF may therefore be mediated by local rather than by peripheral metabolic hormones., (Copyright © 2021 The Authors. Published by Elsevier Inc. All rights reserved.)

The dire need for COVID-19 treatments has inspired strategies of repurposing approved drugs. Amantadine has been suggested as a candidate, and cellular as well as clinical studies have indicated beneficial effects of this drug. We demonstrate that amantadine and hexamethylene-amiloride (HMA), but not rimantadine, block the ion channel activity of Protein E from SARS-CoV-2, a conserved viroporin among coronaviruses. These findings agree with their binding to Protein E as evaluated by solution NMR and molecular dynamics simulations. Moreover, we identify two novel viroporins of SARS-CoV-2; ORF7b and ORF10, by showing ion channel activity in a X. laevis oocyte expression system. Notably, amantadine also blocks the ion channel activity of ORF10, thereby providing two ion channel targets in SARS-CoV-2 for amantadine treatment in COVID-19 patients. A screen of known viroporin inhibitors on Protein E, ORF7b, ORF10 and Protein 3a from SARS-CoV-2 revealed inhibition of Protein E and ORF7b by emodin and xanthene, the latter also blocking Protein 3a. This illustrates a general potential of well-known ion channel blockers against SARS-CoV-2 and specifically a dual molecular basis for the promising effects of amantadine in COVID-19 treatment. We therefore propose amantadine as a novel, cheap, readily available and effective way to treat COVID-19., (© 2021. The Author(s).)