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39 pages, 5261 KB  
Article
Effects of Glucose and Fructose on Production Traits, Organ Weights and Metabolomic Indices in Rats on Different Energy and Nutrient Dense Diets
by József Szabó, Gergely Maróti, Norbert Solymosi, Emese Andrásofszky, András Bersényi, Geza Bruckner and István Hullár
Nutrients 2025, 17(17), 2746; https://doi.org/10.3390/nu17172746 (registering DOI) - 25 Aug 2025
Abstract
The objectives of this study were to determine the dose effect/s of glucose (G) and fructose (F) at different energy densities (ED) of diets on feed intake, body and organ weights, chemical composition of liver, feed conversion, and metabolomic indices (enzymes and hormones). [...] Read more.
The objectives of this study were to determine the dose effect/s of glucose (G) and fructose (F) at different energy densities (ED) of diets on feed intake, body and organ weights, chemical composition of liver, feed conversion, and metabolomic indices (enzymes and hormones). Methods: Seventy-two 10-week-old male Wistar SPF rats were divided into 9 dietary groups and housed individually in metabolic cages. The control group was on a carbohydrate-free high lard diet (L), and for the other 8 treatment groups, the L content of the control diet was gradually replaced by G or F to decrease the dietary ED, in such a way that the nutrients (protein, minerals and vitamins) to energy ratio of the feeds remained constant. These experimental diets were fed to rats for 28 days. Feed intake and body weight were measured twice weekly. On the 28th day of the experiment, the rats were euthanized, and blood and organ samples were collected for further tests. Results and conclusions: The effects of F and G on twenty-six parameters were measured at different EDs of diets. Significant specific F effects (SFE) over the rats on G diets were found in case of feed intake (statistics with pooled data of feed intake (Fi) showed ~7% more feed intake of F rats: 10.8, 6.4, 9.5 and 2.0% at 5.28, 4.70, 4.23 and 3.85 kcal/g ED, respectively); body weight gain (the relation is polynomial; 8.0, 10.3, 0.1, and −10.2% at 5.28, 4.70, 4.23, and 3.85 kcal/g ED; it related to the weight change of viscera: liver, kidney and RWAT); liver fat (3.98, 21.42, 49.20 and 11.05% at 5.28, 4.70, 4.23, and 3.85 kcal/g ED, respectively); serum triglyceride (the relation is polynomial; 63.2, 88.1, 79.2 and 42.6% at 5.28, 4.73, 4.23, and 3.85 kcal/g ED, respectively); serum glucagon (−1.2, 380.2, 248.3 and 74.7% at 5.28, 4.70, 4.23, and 3.85 kcal/g ED, respectively), and serum leptin (9.59, 30.53, 72.64, and −46.49% at 5.28, 4.70, 4.23, and 3.85 kcal/g ED, respectively). An important conclusion is that in several cases, the effects of F and G were similar in the direction of change, but the magnitude of the effects was different. In case of feed conversion rate, there was no difference between the effect of G and F, however it is important to note that the higher the dietary energy and nutrient density, the better the feed conversion rate (FCR); The potential mechanism(s) of effect for each parameter is discussed and, where appropriate, the clinical relevance of the data compared to the known literature. Full article
(This article belongs to the Section Carbohydrates)
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16 pages, 2468 KB  
Article
Targeting the Oviduct Microbiota and Redox Status: A Novel Perspective on Probiotic Use in Laying Hens
by Gabriela Miotto Galli, Ines Andretta, Camila Lopes Carvalho, Aleksandro Schafer da Silva and Marcos Kipper
Poultry 2025, 4(3), 35; https://doi.org/10.3390/poultry4030035 - 7 Aug 2025
Viewed by 366
Abstract
(1) Background: The goal of the present study was to evaluate whether the supplementation with a multi-species probiotic in the diet of laying hens can change the microbiota and health status of the oviduct. (2) Methods: A total of 60 cages housing lightweight [...] Read more.
(1) Background: The goal of the present study was to evaluate whether the supplementation with a multi-species probiotic in the diet of laying hens can change the microbiota and health status of the oviduct. (2) Methods: A total of 60 cages housing lightweight laying hens (36 weeks old) were randomly assigned to the following two different treatments: a control group fed a diet without probiotic, and a treatment group receiving diets supplemented with 50 g/ton of probiotics. The trial lasted for 26 weeks, after which five layers were slaughtered per treatment for oviduct (magnum) assessment, focusing on microbiome composition, oxidant and antioxidant status, and morphological analyses. Additionally, intestinal (jejunum) samples were collected to determine oxidant and antioxidant status. (3) Results: Probiotic supplementation resulted in lower counts of organisms from the RB41 order (p = 0.039) and Burkholderia genus (p = 0.017), and a total reduction in Bacillus and Corynebacterium (p = 0.050) compared to the control treatment. Genera Burkholderia (p = 0.017), Corynebacterium (p = 0.050), and Bacillus (p = 0.050) were also lower with the probiotic supplementation in relation to the control. Genera Epulopiscium (p = 0.089), Flavobacterium (p = 0.100), Ruminococcus (p = 0.089), and Staphylococcus (p = 0.100) tended to be lower in the probiotic group compared to the control. No significant differences were found between treatments for oviduct lesions. Probiotic treatment resulted in a higher protein thiol level in the intestine compared to the control (p < 0.001). However, the use of probiotics tended to reduce glutathione S-transferase levels in the oviduct compared to the control (p = 0.068). (4) Conclusions: These results suggest that dietary supplementation with probiotics can modulate the oviduct microbiota and improve the antioxidant status of laying hens, without causing tissue damage. Further research is warranted to explore the long-term implications of these changes on reproductive performance and egg quality. Full article
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20 pages, 998 KB  
Article
Colony Nutrition Enhances Bee Resilience to Fungicides, While the Benefit of Propolis Supplementation Depends on Stress Conditions
by Yara Martins Molina Ferraz, Aline Yukari Kato, Tainá Angelica de Lima Freitas, Cássia Regina de Avelar Gomes, Thais Regina Ramos Alves, Matheus Franco Trivellato, Samir Moura Kadri, Ricardo de Oliveira Orsi, David De Jong, Jaqueline Dalbello Biller and Daniel Nicodemo
Agriculture 2025, 15(15), 1665; https://doi.org/10.3390/agriculture15151665 - 1 Aug 2025
Viewed by 478
Abstract
Enhanced colony nutrition can support brood development, resulting in better physiological conditions and increased resilience in adult honey bees, particularly under stress. This study investigated the effects of colony nutrition and adult dietary supplementation with green propolis on bee health under fungicide exposure. [...] Read more.
Enhanced colony nutrition can support brood development, resulting in better physiological conditions and increased resilience in adult honey bees, particularly under stress. This study investigated the effects of colony nutrition and adult dietary supplementation with green propolis on bee health under fungicide exposure. Colonies were managed under food restriction or nutritional supplementation for 22 weeks. Newly emerged bees from each colony were then caged and fed protein diets consisting of honey-pollen patties contaminated or not with fungicide, and sucrose sugar syrup with or without aqueous green propolis extract. Bees from supplemented colonies showed greater body weight, higher hemolymph protein levels, and higher consumption of protein food after seven days in cages. Fungicide exposure reduced hemolymph protein levels, altered the expression of detoxification and immune-related genes, and significantly decreased bee survival. Interestingly, propolis supplementation alone changed gene expression patterns and slightly reduced longevity compared to bees not exposed to propolis or fungicide. However, under fungicide stress, bees that ingested propolis survived longer, indicating a protective effect. While colony nutritional supplementation clearly promotes honey bee resilience against fungicide exposure, feeding propolis also showed promising effects, though further studies are needed to determine an optimal dietary concentration. Full article
(This article belongs to the Special Issue Honey Bees and Wild Pollinators in Agricultural Ecosystems)
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13 pages, 652 KB  
Article
Evaluation of Alterations in Nutrient Utilization and Intestinal Health in Response to Heat Stress in Pekin Ducks Based on a Pair-Feeding Experimental Design
by Xiangyi Zeng, Arshad Javid, Gregory S. Fraley, Gang Tian, Keying Zhang, Shiping Bai, Xuemei Ding, Jianping Wang, Yan Liu, Yue Xuan, Shanshan Li and Qiufeng Zeng
Animals 2025, 15(15), 2213; https://doi.org/10.3390/ani15152213 - 28 Jul 2025
Viewed by 355
Abstract
The objective of this study was to investigate alterations in nutrient utilization, standardized ileal digestibility of amino acids (SIDAA), and intestinal health in response to heat stress (HS) in Pekin ducks. A total of 240 healthy 28-day-old male Pekin ducks were randomly allocated [...] Read more.
The objective of this study was to investigate alterations in nutrient utilization, standardized ileal digestibility of amino acids (SIDAA), and intestinal health in response to heat stress (HS) in Pekin ducks. A total of 240 healthy 28-day-old male Pekin ducks were randomly allocated to three groups: a normal control (NC) group, an HS group, and a pair-fed (PF; provided an amount of feed equal to that consumed by the HS group to eliminate the effects of feed intake) group, each with eight replicate cages of ten birds. The results showed that HS significantly reduced the apparent utilization of dietary energy, ether extract, and crude protein compared to both the NC and PF groups (p < 0.05), but yielded comparable SIDAA to the PF group. The HS group exhibited reduced mRNA levels of EAAT3 and PepT1, along with elevated mRNA levels of CAT1, GLUT5, and FATP6 in the jejunum compared to the NC or PF groups, respectively (p < 0.05). Furthermore, HS resulted in a significant deterioration of jejunal morphology and goblet cell count compared to the NC and PF groups (p < 0.05). Serum fluorescein isothiocyanate-dextran levels were significantly higher in HS ducks than in NC ducks (p < 0.05), but did not differ from PF ducks. At order-level classification of ileal mucosal microbiota, HS markedly increased the relative abundance of Bacillales, Deferribacterales, and Actinomycetales versus NC (p < 0.05), while significantly decreasing Bifidobacteriales abundance relative to PF (p < 0.05). Collectively, HS induces a leaky gut and microbiota dysbiosis that compromises gut health, thereby reducing dietary nutrient utilization in Pekin ducks. The observed reduction in feed intake constitutes a primary driver of intestinal health deterioration in heat-stressed Pekin ducks. Full article
(This article belongs to the Section Poultry)
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21 pages, 3372 KB  
Article
Advanced Research on Biological Properties—A Study on the Activity of the Apis mellifera Antioxidant System and the Crystallographic and Spectroscopic Properties of 7-Diethylamino-4-hydroxycoumarin
by Klaudia Rząd, Iwona Budziak-Wieczorek, Aneta Strachecka, Patrycja Staniszewska, Adam Staniszewski, Anna Gryboś, Alicja Matwijczuk, Bożena Gładyszewska, Karolina Starzak, Anna A. Hoser, Maurycy E. Nowak, Małgorzata Figiel, Sylwia Okoń and Arkadiusz Paweł Matwijczuk
Int. J. Mol. Sci. 2025, 26(14), 7015; https://doi.org/10.3390/ijms26147015 - 21 Jul 2025
Viewed by 562
Abstract
The search for substances that increase the immunity of bees is becoming a necessity in the era of various environmental threats and the declining immunocompetence of these insects. Therefore, we tested the biological and physicochemical properties of 7-diethylamino-4-hydroxycoumarin (7DOC). In a cage test, [...] Read more.
The search for substances that increase the immunity of bees is becoming a necessity in the era of various environmental threats and the declining immunocompetence of these insects. Therefore, we tested the biological and physicochemical properties of 7-diethylamino-4-hydroxycoumarin (7DOC). In a cage test, two groups of bees were created: a control group fed with sugar syrup and an experimental group fed with sugar syrup with the addition of 7DOC. In each group, the longevity of the bees was determined and the protein concentrations and antioxidant activities in the bees’ hemolymph were determined. The bees fed with 7DOC lived 2.7 times longer than those in the control group. The protein concentrations and activities of SOD, CAT, GPx and GST, as well as the TAC levels, were significantly higher in the hemolymph of the supplemented workers. To confirm these potent biological properties of 7DOC, the UV-Vis spectra, emission and excitation of fluorescence, synchronous spectra and finally the fluorescence lifetimes of this compound were measured using the time-correlated single photon counting method, in various environments differing in polarity and in the environment applied in bee research. This compound was shown to be sensitive to changes in solvent polarity. The spectroscopic assays were complemented with crystallographic tests of the obtained monocrystals of the aforementioned compounds, which attested to the aggregation effects observed in the spectra measurements for the selected coumarin. The research results confirm that this compound has the potential to be implemented in apiary management, which will be our application goal, but further research into apiary conditions is required. Full article
(This article belongs to the Section Bioactives and Nutraceuticals)
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27 pages, 4515 KB  
Article
Effects of Different Farming Models on Muscle Quality, Intestinal Microbiota Diversity, and Liver Metabolism of Rice Field Eel (Monopterus albus)
by Yifan Zhao, Wenzong Zhou, Muyan Li, Yuning Zhang, Weiwei Lv, Weiwei Huang, Hang Yang, Quan Yuan and Mingyou Li
Foods 2025, 14(13), 2383; https://doi.org/10.3390/foods14132383 - 5 Jul 2025
Viewed by 643
Abstract
As consumer demand for quality fish products continues to rise, quality has become a key factor in market competition. Ecological aquaculture research is exploring various farming methods to balance high-quality demand with environmental protection. This study compared three aquaculture models—cage culture (CG), recirculating [...] Read more.
As consumer demand for quality fish products continues to rise, quality has become a key factor in market competition. Ecological aquaculture research is exploring various farming methods to balance high-quality demand with environmental protection. This study compared three aquaculture models—cage culture (CG), recirculating aquaculture (RAG), and rice–fish co-culture (RG)—by analyzing muscle quality (AOAC, GC-MS), intestinal microbiota (16S rRNA), and liver metabolism (LC-MS) to assess their effects on M. albus. In terms of muscle quality, the RG group showed increased levels of EPA and DHA, reduced muscle moisture and crude lipid content, and enhanced crude protein accumulation. The crude protein content was significantly higher in the RAG group than in the CG group (p < 0.05). The RG group also had the highest levels of total, essential, and umami amino acids, followed by the RAG and CG groups. In terms of intestinal microbiota, the RG group had the highest microbial diversity and stability, with increased abundance of Firmicutes and Bacteroidetes and decreased levels of Proteobacteria. Compared to the CG, the RAG group also showed increased microbial diversity and a reduction in pathogenic genera. Liver metabolomics analysis demonstrated that the RG group had significant advantages over the CG group in amino acid, lipid, and energy metabolism. The RAG group exhibited upregulation of glycerophospholipid metabolism and a decrease in oxidative stress marker levels. Overall, the RG group enhanced muscle quality and optimized intestinal and liver metabolism in M. albus. Full article
(This article belongs to the Section Meat)
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24 pages, 8724 KB  
Article
Transcriptomic Analysis of Trachinotus ovatus Under Flow Velocity Stress
by Jing Zhang, Xixi Liu, Jiayue Dai, Sufang Niu, Xuefeng Wang and Baogui Tang
Animals 2025, 15(13), 1932; https://doi.org/10.3390/ani15131932 - 30 Jun 2025
Viewed by 365
Abstract
Trachinotus ovatus is a euryhaline, warm-water pelagic fish species with strong adaptability, rapid growth, and a high survival rate, making it one of the most important marine aquaculture species in China. In recent years, extensive experience has been accumulated in the cage farming [...] Read more.
Trachinotus ovatus is a euryhaline, warm-water pelagic fish species with strong adaptability, rapid growth, and a high survival rate, making it one of the most important marine aquaculture species in China. In recent years, extensive experience has been accumulated in the cage farming of T. ovatus, but whether it can adapt to deep-sea environments and grow normally remains a current research focus. This study used RNA-Seq sequencing technology to analyze the gene expression changes in the liver of T. ovatus under three conditions: rest (0 cm/s), medium flow velocity (54 cm/s), and high flow velocity (90 cm/s). Through differential expression analysis, Short Time-series Expression Miner (STEM) analysis and protein–protein interaction (PPI) network analysis, a total of 5107 differentially expressed genes (DEGs), three significantly expressed gene profiles (profile6, profile1, and profile5), and 15 hub genes were identified. The results showed that changes in flow speed significantly impacted key biological processes such as energy metabolism, protein homeostasis, and endoplasmic reticulum (ER) stress response. Under moderate and high flow conditions, glycolysis-related genes were upregulated to meet the energy demands of swimming, while the downregulation of the PPARγ-RXRG complex and its downstream genes in the lipid metabolism pathway suggested a limitation in its fatty acid β-oxidation capacity. At the same time, protein synthesis was enhanced, and the unfolded protein response (UPR) was activated to help cope with ER stress. Furthermore, when the flow speed reached 90 cm/s, the expression of UPR- related genes and the anti-apoptotic factor JNK significantly decreased, suggesting that the stress response was nearing its limit and could potentially trigger cell apoptosis. These findings provide new insights into the molecular adaptation mechanisms of T. ovatus to flow speed stress and offer theoretical support for its rational farming in deep-sea cages, suggesting that the water flow speed in farming should not exceed 90 cm/s. Full article
(This article belongs to the Section Aquatic Animals)
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19 pages, 2192 KB  
Article
Transcriptome Analysis of the Hippocampus in Domestic Laying Hens with Different Fear Responses to the Tonic Immobility Test
by Jingyi Zhang, Min Li, Liying Pan, Ye Wang, Hui Yuan, Zhiwei Zhang, Chaochao Luo and Runxiang Zhang
Animals 2025, 15(13), 1889; https://doi.org/10.3390/ani15131889 - 26 Jun 2025
Viewed by 372
Abstract
Fear is a critical welfare concern in laying hens. Fearful behaviors in domestic chickens are influenced by both genetic and environmental factors, contributing to individual differences in stress responses. Tonic immobility (TI) duration is widely recognized as a reliable indicator of fear levels. [...] Read more.
Fear is a critical welfare concern in laying hens. Fearful behaviors in domestic chickens are influenced by both genetic and environmental factors, contributing to individual differences in stress responses. Tonic immobility (TI) duration is widely recognized as a reliable indicator of fear levels. The hippocampus, a critical brain region for emotional states, plays a pivotal role in associating fearful experiences with specific stimuli, enabling adaptive behavioral responses. This study investigated hippocampal histological characteristics and transcriptomic profiles in laying hens with different fear responses categorized based on TI duration. A total of 80 native Lindian hens (75 weeks old) were individually housed in modified conventional cages. At 76 weeks of age, hens exhibiting the longest and shortest TI durations were classified into the high-fear (TH) and low-fear (TL) groups, respectively. Whole hemibrains were collected for histological and immunohistochemical analyses, while hippocampal tissues underwent transcriptome sequencing. The results showed a significant reduction in Nissl body counts in hippocampal neurons of high-fear hens (p < 0.05), suggesting potential neuronal damage or functional impairment. Transcriptomic analysis revealed 365 differentially expressed genes (DEGs) between two groups, with 277 upregulated and 88 downregulated genes in TH chickens. KEGG pathway enrichment analysis identified seven significantly associated pathways (p < 0.01), including retinol metabolism, vitamin B6 metabolism, and nicotinate and nicotinamide metabolism, all of which are crucial for neuronal function and immune regulation. In addition, a significant increase in DCX protein expression (p < 0.05) and a decrease in c-Fos protein expression (p < 0.05) was noted in in high-fear hens, whereas PCNA levels remained unchanged (p > 0.05) under immunohistochemical validation. The neuronal alterations observed in high fear individuals suggest neural damage, while transcriptomic variations point to potential disruptions in neurogenesis, synaptic signaling, and stress-related pathways. Collectively, these results provide novel insights into the neurobiological basis of fear regulation in laying hens and may have implications for poultry welfare and management strategies. Full article
(This article belongs to the Special Issue Welfare and Behavior of Laying Hens)
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25 pages, 554 KB  
Article
Assessment of the Effects of Stocking Density on Laying Hens Raised in Colony Cages: Part II—Egg Production, Egg Quality, and Welfare Parameters
by Benjamin N. Alig, Kenneth E. Anderson, Dimitri M. Malheiros, Kari L. Harding and Ramon D. Malheiros
Poultry 2025, 4(3), 28; https://doi.org/10.3390/poultry4030028 - 20 Jun 2025
Viewed by 833
Abstract
Stocking density is one of the major concerns in all production systems, which is why lawmakers, retailers, and consumers are increasingly concerned about this issue and its relation to animal welfare. The aim of this study was to identify if stocking density had [...] Read more.
Stocking density is one of the major concerns in all production systems, which is why lawmakers, retailers, and consumers are increasingly concerned about this issue and its relation to animal welfare. The aim of this study was to identify if stocking density had an effect on white egg layer production, egg quality, bird health, and welfare parameters. For this study, five stocking densities were evaluated in colony cages: 1342 cm2/hen, 897 cm2/hen, 671 cm2/hen, 535 cm2/hen, and 445 cm2/hen. Egg production and physical egg quality were measured. Hen health and welfare parameters including corticosterone levels, H/L, oxidative stress pathways, jejunum tight junction protein expression, bone health, gut histology, body condition, and cytokine expression were evaluated. The results from this indicated that higher stocking densities resulted in lower production but larger eggs, while feed efficiency remained unaffected. Furthermore, physical egg quality parameters also remained unaffected. When stress and welfare parameters were analyzed, this research identified that decreasing stocking density did not affect jejunum oxidative stress pathways, pro-inflammatory cytokine expression, bone health, or intestinal health. At the final sampling period (69 weeks), the highest stocking density demonstrated higher corticosterone concentrations and IL-10 expression compared to the lowest stocking density. Furthermore, feather scores were found to be poorer as density increased. Finally, the highest density had higher Hansen’s test scores compared to other densities, which indicates a greater fear response. In conclusion, it appears that decreasing stocking density may provide some benefits in production and welfare to commercial egg layers, particularly at the end of the laying cycle. Full article
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18 pages, 1517 KB  
Article
Renal and Vascular Effects of the Allosteric Transglutaminase 2 Modulator LDN-27219 in One-Kidney DOCA–Salt Mice
by Ian Mees, Judit Prat-Duran, Simon Comerma-Steffensen, Ulf Simonsen, Estéfano Pinilla and Niels Henrik Buus
Int. J. Mol. Sci. 2025, 26(12), 5724; https://doi.org/10.3390/ijms26125724 - 14 Jun 2025
Viewed by 656
Abstract
The enzyme transglutaminase 2 (TG2) has an open conformation with transamidase activity which crosslinks matrix proteins contributing to fibrosis development. LDN-27219 promotes the closed conformation of TG2, which can enhance vasodilation, but its effects in renal tissue are unknown. We investigated whether LDN-27219 [...] Read more.
The enzyme transglutaminase 2 (TG2) has an open conformation with transamidase activity which crosslinks matrix proteins contributing to fibrosis development. LDN-27219 promotes the closed conformation of TG2, which can enhance vasodilation, but its effects in renal tissue are unknown. We investigated whether LDN-27219 treatment affects albuminuria and markers of renal fibrosis as well as ex vivo vasodilatation. Male C57BL/6 mice (n = 48) underwent unilateral nephrectomy plus insertion of a deoxycorticosterone acetate pellet (DOCA group) or nephrectomy only (sham group). Both groups were randomized to intraperitoneal treatment with either LDN-27219 (8 mg/kg twice daily) or vehicle for 2 weeks. Urine albumin excretion was evaluated by metabolic cages. Kidney tissue fibrosis markers were assessed by qPCR and Western blotting, while the TG2 conformational state was evaluated using native gel electrophoresis. Collagen staining was performed using Picrosirius red and quantified under circularly polarized light. Mesenteric arteries were mounted in wire myographs for evaluation of vasorelaxation. DOCA mouse developed significant albuminuria (p < 0.001 vs. sham), but neither TG2 mRNA nor protein expression was upregulated in the kidney. However, the relative amount of TG2 in the closed conformation was higher in DOCA mice. LDN-27219 did not affect albuminuria, but LDN-27219-treated DOCA mice showed less urine production and less collagen staining than vehicle-treated DOCA mice. LDN-27219 did not affect TG2 mRNA or TG2 protein expression or mRNA of fibrosis markers. LDN-27219-treated mice had enhanced vasorelaxation to the nitric oxide donor sodium nitroprusside. In conclusion, LDN-27219 treatment in the one-kidney DOCA–salt model did not affect renal TG2 mRNA and protein expression or albuminuria but still exerted beneficial effects in terms of reduced kidney fibrosis and urine production in addition to enhanced vasodilatation. Full article
(This article belongs to the Special Issue The Latest Molecular Research on Renal Disease)
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23 pages, 925 KB  
Article
Bi-Symmetric Polyhedral Cages with Nearly Maximally Connected Faces and Small Holes
by Bernard Piette
Symmetry 2025, 17(6), 940; https://doi.org/10.3390/sym17060940 - 12 Jun 2025
Cited by 1 | Viewed by 611
Abstract
Polyhedral cages (p-cages) provide a good description of the geometry of some families of artificial protein cages. In this paper we identify p-cages made out of two families of equivalent polygonal faces/protein rings, where each face has at least four neighbours and where [...] Read more.
Polyhedral cages (p-cages) provide a good description of the geometry of some families of artificial protein cages. In this paper we identify p-cages made out of two families of equivalent polygonal faces/protein rings, where each face has at least four neighbours and where the holes are contributed by at most four faces. We start the construction from a planar graph made out of two families of equivalent nodes. We construct the dual of the solid corresponding to that graph, and we tile its faces with regular or nearly regular polygons. We define an energy function describing the amount of irregularity of the p-cages, which we then minimise using a simulated annealing algorithm. We analyse over 600,000 possible geometries but restrict ourselves to p-cages made out of faces with deformations not exceeding 10%. We then present graphically some of the most promising geometries for protein nanocages. Full article
(This article belongs to the Special Issue Chemistry: Symmetry/Asymmetry—Feature Papers and Reviews)
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15 pages, 1915 KB  
Communication
Performance of Imidazoquinoline Glycoconjugate BAIT628 as a TLR7 Agonist Prodrug for Prostate Cancer
by Seyedeh A. Najibi, S. M. Al Muied Pranto, Muhammad Haroon, Amy E. Nielsen and Rock J. Mancini
Pharmaceuticals 2025, 18(6), 804; https://doi.org/10.3390/ph18060804 - 27 May 2025
Viewed by 1013
Abstract
Despite broad anti-cancer efficacy as Toll-Like Receptor (TLR) 7/8 agonists, imidazoquinolines remain limited in use via systemic administration or in situ vaccination therapies due to inflammatory toxicity. One approach to address this challenge involves better targeting the action of imidazoquinolines by caging them [...] Read more.
Despite broad anti-cancer efficacy as Toll-Like Receptor (TLR) 7/8 agonists, imidazoquinolines remain limited in use via systemic administration or in situ vaccination therapies due to inflammatory toxicity. One approach to address this challenge involves better targeting the action of imidazoquinolines by caging them as glycoconjugate prodrugs. Within cancer cells, imidazoquinoline glycoconjugates are activated by hydrolases prior to efflux by ABC transport proteins, where they then elicit tumoricidal effects from the assistance of bystander immune cells, such as tumor-infiltrating lymphocytes and associated macrophages, in local proximity. While this concept of Bystander-Assisted ImmunoTherapy (BAIT) has been established at a molecular level in vitro, tolerability or efficacy of BAIT has not been reported in vivo. Here, we evaluate the MTD and tumor growth delay efficacy of a lead BAIT prodrug (BAIT628) in a male C57BL/6 mouse TRAMP-C2 prostate cancer model to further establish this methodology. Overall, we find that systemic BAIT628 is well tolerated at over 5-fold the dose-limiting inflammatory toxicity of the parent imidazoquinoline (up to 5 mg/mouse/day I.P. for 10 days). Analyzing serum cytokines reveals that IL-10 production, elicited by the mannoside caging group, likely contributes to the enhanced MTD. Using BAIT628 as an in situ vaccination immunotherapy (seven times over 3 weeks) resulted in significant tumor growth delay and increased survival, both alone and in combination with a murinized α-PD-L1 checkpoint blockade. The tumor histology of tumor-infiltrating immune cell subsets (CD4+, CD8+, CD11c+) reveals significant increases in CD11c+ populations, consistent with TLR7/8 agonism. Overall, BAIT628 is well tolerated and exhibits significant efficacy in the TRAMP-C2 model. These results demonstrate how the BAIT approach can optimize imidazoquinolines for in vivo tolerability and subsequent efficacy as cancer immunotherapeutics. Full article
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21 pages, 9809 KB  
Article
Genome-Wide Identification and Expression Analysis of the YTH Domain-Containing Protein Gene Family in Salvia miltiorrhiza
by Chunling Wang, Yunliang Peng, Xian Pan, Sixuan Zhang, Yayun Xu, Caili Li, Butuo Zhu, Lili Niu and Shanfa Lu
Int. J. Mol. Sci. 2025, 26(10), 4645; https://doi.org/10.3390/ijms26104645 - 13 May 2025
Viewed by 586
Abstract
YTH domain-containing proteins act as the primary readers of N6-methyladenosine (m6A), playing an important role in plant development and stress responses. However, little is known about the YTH proteins in medicinal plants. Genome-wide identification of the YTH gene family [...] Read more.
YTH domain-containing proteins act as the primary readers of N6-methyladenosine (m6A), playing an important role in plant development and stress responses. However, little is known about the YTH proteins in medicinal plants. Genome-wide identification of the YTH gene family in the medicinal model plant, Salvia miltiorrhiza Bunge, identified a total of nineteen SmYTH genes from five chromosomes, with SmYTH8SmYTH19 clustered on chromosome 8. Phylogenetic analysis showed that SmYTH proteins belong to the YTHDF category. No YTHDC members were identified. Conserved domain identification, amino acid sequence alignment, and phase separation prediction revealed that SmYTH1–SmYTH4 exhibited the characteristic m6A reader protein feature, containing conserved aromatic cages (WWW) capable of binding m6A residues. SmYTH5–SmYTH19 proteins contain a unique conserved F-box protein interaction domain that has not been reported previously. qRT-PCR analysis revealed tissue-specific patterns, with SmYTH1SmYTH4 genes highly expressed in roots and leaves, whereas SmYTH8SmYTH19 were mainly expressed in leaves. The results were consistent with RNA-seq data. The expression of various SmYTHs and the content of phenolic acid active ingredients were significantly altered under MeJA and SA treatments. The results provide useful information for further studies on the biological functions of m6A and YTH proteins in S. miltiorrhiza. Full article
(This article belongs to the Section Molecular Plant Sciences)
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23 pages, 23181 KB  
Article
Ultrasound-Assisted Ferritin Extraction from Northern Pike Liver: An Innovative Approach for Chlorogenic Acid Encapsulation with Enhanced Thermal Stability
by Zhikun Xing, Yi Wang, Yabo Wei, Xin Guo, Xiaoyue Liang, Xiaorong Deng, Lianfu Zhang and Jian Zhang
Molecules 2025, 30(9), 2080; https://doi.org/10.3390/molecules30092080 - 7 May 2025
Cited by 1 | Viewed by 573
Abstract
Ferritin, an emerging protein resource, has garnered significant attention in scientific research due to its biocompatibility and unique cavity structure capable of encapsulating bioactive compounds. This study aimed to optimize ultrasound-assisted extraction (UAE) for enhancing ferritin yield from northern pike liver byproducts and [...] Read more.
Ferritin, an emerging protein resource, has garnered significant attention in scientific research due to its biocompatibility and unique cavity structure capable of encapsulating bioactive compounds. This study aimed to optimize ultrasound-assisted extraction (UAE) for enhancing ferritin yield from northern pike liver byproducts and evaluate its potential as a nanocarrier for chlorogenic acid (CA). Through response surface methodology (RSM), the optimal UAE parameters were established as 200 W ultrasonic power, 1:3 solid–liquid ratio, and 25 min extraction time. Under these conditions, the ferritin extraction yield reached 139.46 mg/kg, representing a 4.02-fold increase compared to conventional methods (34.65 mg/mL). Electrophoretic analysis confirmed the electrophoretic purity of the extracted liver ferritin. Comprehensive characterization using UV-vis spectroscopy, FTIR, and fluorescence spectroscopy revealed preserved structural integrity of UAE-extracted ferritin. Homology modeling provided molecular insights into the ferritin architecture. Successful encapsulation of CA was achieved with an encapsulation efficiency of 13.25%, as quantified by HPLC. Analysis by DLS and ζ potential as well as TG and DSC showed that not only the thermal stability of CA was enhanced after ferritin encapsulation, but also that the ferritin remained stable with a cage-like structure. This investigation establishes UAE as an effective strategy for valorizing fish processing byproducts through high-yield ferritin extraction while demonstrating the protein’s functional capacity as a nanocarrier for bioactive compound delivery. The findings highlight the dual advantage of sustainable resource utilization and advanced delivery system development through this biotechnological approach. Full article
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Article
Effect of Berchemia discolor Leaf Meal (Muni Tree) on Feed Intake, Growth Performance and Digestibility of Non-Descript Goats
by Jobere Anastacia Mashiachidi, Tlou Grace Manyelo and Busisiwe Gunya
Animals 2025, 15(9), 1342; https://doi.org/10.3390/ani15091342 - 6 May 2025
Viewed by 446
Abstract
Seasonal feed shortages often limit goat productivity in rural areas, creating a need for alternative, locally available feed resources. This study evaluated the effect of Berchemia discolor leaf meal on feed intake, digestibility and growth performance of South African non-descript goats. Sixteen yearling [...] Read more.
Seasonal feed shortages often limit goat productivity in rural areas, creating a need for alternative, locally available feed resources. This study evaluated the effect of Berchemia discolor leaf meal on feed intake, digestibility and growth performance of South African non-descript goats. Sixteen yearling goats (average age 12 months, body weight 19.63 ± 1.68 kg) were randomly assigned to four dietary treatments in a completely randomized design and housed individually in metabolic cages. They were fed diets containing 0%, 15%, 20% and 30% B. discolor leaf meal, formulated on a dry matter basis for 42 days (14-day adaptation, 28-day measurement). B. discolor supplementation had no effect on weight and feed intake (p > 0.05). However, goats fed diets with 0% and 30% inclusion levels had a significantly higher average daily gain and more efficient feed conversion ratio compared to those fed 15% and 20% inclusion levels (p < 0.05). Daily dry matter intake did not differ (p > 0.05) among groups. However, B. discolor had lower (p < 0.05) crude protein (CP), neutral detergent fiber (NDF) and acid detergent fiber (ADF) contents compared to the control (0% inclusion). Metabolic weight intakes were similar (p > 0.05) across treatments. B. discolor had no effect (p > 0.05) on dry matter digestibility, but CP and ADF digestibility were higher (p < 0.05) in goats fed 0% B. discolor than those in 15%, 20% or 30% inclusion groups. In conclusion, B. discolor inclusion improved growth performance and feed conversion efficiency in non-descript goats, though higher levels reduced nutrient digestibility. Inclusion up to 20% supported intake and growth, but the highest feed conversion ratio was observed at 0% and 30% inclusion. Full article
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