Tail regeneration in lizards occurs after formation of a wound (regenerating) epidermis that surrounds and protects the inner mass of mesenchymal-like cells of regenerating blastema formed in highly hydrated conditions. Aside mechanical protection, enrichment of lipids mixed with Intermediate Filament Keratins (IFKs) is essential for maintaining the blastema alive to form a regenerated tail. After tail amputation in the wall lizard Podarcis muralis, a regenerating wound epidermis is formed which keratinocytes accumulate lipids among IFKs during their differentiation into corneocytes. The present mass spectrometric study characterizes in details the different lipids produced in normal and regenerating tail epidermis of this reptile. Among the classes of lipids that undergo to significative variations are included cholesterol esters and hexosylceramides that increase in comparison to the normal, non-regenerating epidermis. Among other lipid classes, also lysophospholipids, phosphoglycerids and cardiolipins undergo a significant increment in wound keratinocytes, probably related to cell membrane turnover during keratinocyte regeneration. In contrast, in wound keratinocytes triglycerides decrease in comparison to those of normal epidermis. Other classes of lipids do not differ significantly between normal and regenerating epidermis, indicating minor roles for the formation of a water-loss barrier. We suggest that the marked changes in lipid profile due to the increase in cholesteroyl-esters and hexosylceramides in parallel with the triacylglycerols decrease, are correlated with the establishment of a more effective barrier against water loss in the lizard epidermis. The evolution of an efficient barrier to water-loss was essential, in addition to specific gene developmental pathways, for tail regeneration in lizards.

This study evaluated the effects of partially replacing soybean meal (SBM) with a combination of cricket meal (CM; Acheta domesticus nymph meal) and Moringa oleifera leaf meal (MOLM) on growth performance, carcass characteristics, and apparent ileal nutrient digestibility in broiler chickens. A total of 240 one-day-old unsexed broilers were assigned to four dietary treatments with six replicate pens of ten birds each. Treatments consisted of a control diet (D0) and three diets in which 10% (D1), 20% (D2), or 30% (D3) of the SBM inclusion level was replaced by a cm-MOLM blend at a 3:1 ratio. Titanium dioxide was used as an indigestible marker to determine digestibility. Broilers fed D1 and D2 showed greater body weight gain (BWG) than those fed D0 and D3. Feed conversion ratio (FCR) was not significantly affected (P = 0.089), although birds fed D3 had lower growth despite higher feed intake (FI). Carcass weight and thigh yield were higher in D1 and D2 compared with the control (P < 0.001). Apparent ileal digestibility of crude protein and organic matter decreased at the 30% substitution level (P < 0.001), whereas ether extract digestibility increased with higher inclusion of the blend. Relative weights of the gizzard and liver increased in D3. Serum total cholesterol, triglycerides, and LDL-cholesterol decreased with increasing substitution levels (P < 0.001), while HDL-cholesterol increased. In conclusion, replacing up to 20% of SBM with a cm-MOLM blend improved growth performance, carcass traits, and lipid profiles without impairing nutrient digestibility. These findings suggest that a 20% substitution level can be effectively utilized as a sustainable alternative protein source to reduce reliance on SBM while enhancing lean meat yield and the lipid profile of broilers.

The current study investigated the impact of supplementing polyphenols extracted from shredded, steam-exploded pine particles (PSPP) on the performance, gene expression, and gut metagenome of broilers exposed to cyclic heat stress (CHS). A total of 216 chickens were distributed into a 2 (temperature) by 3 (diets) design, with each treatment consisting of six replicates of six chickens. Specifically, chickens were fed diets containing 0% PSPP, 0.5% PSPP, and 1% PSPP and exposed to two temperature conditions: CHS (31°C) and Thermoneutral (NT, 21°C). The CHS was conducted for 6 hours every day for 7 consecutive days. Final body weight, average daily gain, and average daily feed intake (ADFI) were decreased, while feed conversion ratio and rectal temperature were increased in heat-exposed chickens. Dietary PSPP supplementation enhanced ADFI. The weight of the liver, bursa, and length of the jejunum and ileum were decreased in heat-exposed chickens. Plasma cholesterol was increased, and triglycerides were decreased in heat-exposed chickens. After heat exposure, gene expression of ZO1, ZO2, GLP2, NOX1, SOD, GPX, HSP70, HSP90, NRF2, TLR2, and TLR4 increased in the jejunum. GLP2 gene expression was similar in 1%PSPP exposed to HS in comparison to the entire NT-exposed chickens. Concerning microbiota analysis, alpha diversity indices, such as Shannon and Gini-Simpson, were increased following CHS exposure. Beta diversity, measured through unweighted and weighted UniFrac distances, showed temperature, dose, and interaction effects. The relative abundance of the phylum Candidatus Melainabacteria was increased, while Tenericutes populations were decreased in heat-exposed chickens. Furthermore, a total of thirty genera were identified as microbial biomarkers of CHS. Interestingly, the relative abundance of five pathogenic bacterial genera was found to be decreased in the 0.5%PSPP treatment. Overall, CHS negatively influences growth performance, modulates the expression of the gut antioxidant-related genes, and favors the colonization of pathogenic bacteria. However, 0.5% PSPP may mitigate CHS by reducing pathogen colonization in the gut of broilers.

This study investigates how adding letrozole (LE) to the diet affects the growth, blood hormones and metabolites, rumen fermentation, and gut microbes of weaned female Turpan Black sheep. The results could help guide strategies to enhance female growth and open new avenues for research. The study selected 60 weaned female lambs of the Turpan Black sheep breed, aged 2 months, with an average body weight of (19.93 ± 0.46) kg and in good health, which were randomly divided into the Con group (baseline diet), the 0.1 LE group (baseline diet + 0.10 mg/kg BW-1), and the 0.2 LE group (baseline diet + 0.20 mg/kg BW-1), with 20 lambs in each group. The pre-feeding period lasted 5 days, and the main feeding period lasted 75 days. The results showed that 0.2 LE group significantly improved total weight gain, body length and chest girth (P < 0.05), and markedly increased plasma GH and IGF-1 concentrations (P < 0.01). On days 15 and 75, plasma LE concentration in the 0.2 LE group was extremely higher than that in the 0.1 LE group (P < 0.01). Metabolomic analysis indicated that differential metabolites in the 0.1 LE group were mainly enriched in bile acid metabolism, with significant upregulation of estrone, taurocholic acid and fumaric acid (P < 0.05); while the 0.2 LE group presented altered cholesterol metabolism, accompanied by increased creatine and letrozole levels but decreased estrone and fumaric acid (P < 0.05). After 75 days of feeding, ruminal LE concentration in the 0.2 LE group was significantly higher than that in the 0.1 LE group (P < 0.05). All LE supplementation groups had higher rumen volatile fatty acid contents than the control group (P < 0.05), and the acetate-propionate ratio was higher in the 0.1 LE group relative to the 0.2 LE group (P < 0.05). LE treatment increased Firmicutes and decreased Bacteroidetes abundance. Within Firmicutes, the families Lachnospiraceae and Eubacteriaceae were higher in relative abundance. The LEfSe analysis showed that Butyrivibrio fibrisolvens was significantly enriched in the 0.1 LE group (P < 0.05), while Oribacterium was significantly enriched in the 0.2 LE group (P < 0.05). In conclusion, LE supplementation increased ruminal Lachnospiraceae and Erysipelotrichaceae, elevated butyrate, propionate and other VFAs. These products together with LE modulated bile acid/cholesterol metabolism and plasma hormones, supporting lamb growth. The optimal dose was 0.20 mg/kg BW-1.

Polycystic ovary syndrome (PCOS) is a complex endocrine disorder. Exercise is one of the interventions that leads to PCOS improvement. This study aimed to investigate the potential of different durations of high-intensity interval training (HIIT) or continuous training (CT) with/ without metformin (MET) in a rat model of PCOS. For this purpose, the rats were randomly placed in seven groups, including control, PCOS, PCOS + CT, PCOS + HIIT, PCOS + MET, PCOS + MET + CT, and PCOS + MET + HIIT. The PCOS was induced by a single intra-muscular injection of 2.00 mg kg-1 estradiol valerate. The concentration of lipid indices showed that serum cholesterol and low-density lipoprotein decreased, and high-density lipoprotein increased with 8 and 12 weeks of exercise training in PCOS + CT, PCOS + HIIT, PCOS + MET + CT, and PCOS + MET + HIIT groups compared to the PCOS group, but no significant difference was found between groups. Triglyceride level decreased in treated groups with 12 weeks of exercise compared to the PCOS group, and there was no significant difference between the treated groups. Finally, serum malondialdehyde level decreased in groups received exercise intervention alone or combined with MET compared to the PCOS group. These findings revealed that HIIT and CT with longer periods with/without MET can lead to improvement of lipid profile and malondialdehyde level of PCOS patients.

Subterranean burrows of plateau zokors (Eospalax baileyi) on the Qinghai-Tibet Plateau become significantly colder and more hypoxic with increasing altitude. Because cholesterol biosynthesis is highly oxygen-intensive, it is typically inhibited by hypoxia. Here, our findings revealed that with increasing altitude, the expression and activation of SREBP2, alongside FDFT1 and SQLE expression, were significantly upregulated in the liver. Furthermore, the incomplete degradation of SQLE produced two distinct isoforms. High-altitude zokors showed an increase in the active trunSQLE-1 and a concomitant decrease in the inactive trunSQLE-2, completely opposite to low-altitude zokors. Additionally, blood cholesterol and oxygen consumption increased significantly, with no significant difference in arterial oxygen saturation. Notably, body weight changes of plateau zokors conformed to Bergmann's rule. These results suggested that the elevated cholesterol synthesis was not hypoxia-induced. Therefore, the increase in cholesterol synthesis in plateau zokors likely results from adaptation to the cold and humid underground burrow habitat.

Etoxazole (ETX) is a widely used organofluorine acaricide/insecticide applied to many food crops, and its residues have been detected in food matrices at levels of concern for human dietary exposure. However, its hepatotoxic and nephrotoxic potential following subchronic oral exposure remains insufficiently characterised. This study investigated the dose-dependent effects of repeated oral ETX administration on serum biochemical parameters, hepatic and renal antioxidant enzyme activities, lipid peroxidation, and histopathological features in Wistar albino rats. Six groups of seven female rats each received normal control (NC) or ETX at 25, 100, 250, 500, or 750 mg/kg/day by gavage for 28 consecutive days. At the end of the exposure period, serum parameters (AST, ALT, ALP, GGT, creatinine, uricacid, glucose, total cholesterol, triglycerides, HDL, LDL), catalase (CAT), glutathione S-transferase (GST), reduced glutathione (GSH), and malondialdehyde (MDA) levels in liver and kidney tissues, and histopathological changes were evaluated. ETX caused significant, dose-dependent increases in AST and ALT from ETX100, in ALP and GGT from ETX500, and in creatinine and uric acid at higher doses, indicating progressive hepatic and renal injury. Hepatic and renal CAT, GST, and GSH activities significantly decreased, whereas MDA levels significantly increased in the higher-dose groups, demonstrating oxidative stress-mediated disruption of antioxidant defences. Histopathological evaluation revealed dose-dependent liver injury and significant tubular damage in kidneys. The tentative NOAEL and LOAEL were estimated at 25 and 100 mg/kg/day, respectively. These findings demonstrate that ETX induces dose-dependent hepatorenal toxicity in rats through an oxidative stress-mediated mechanism and provide relevant data for dietary risk assessment.

The global escalation of antimicrobial resistance (AMR) necessitates innovative therapeutic interventions. Methicillin-resistant Staphylococcus aureus (MRSA) remains a primary clinical threat due to its extensive resistance profile. In this study, CRISPR-Cas9 technology was employed to target the ermA gene, which is responsible for erythromycin resistance in clinical MRSA isolates. A cationic liposome (Lip) composed of cholesterol, DOTAP, and DOPE was developed to deliver the pCasSA plasmid. Particle characterization revealed stable liposomes (Lips) (<200 nm, PDI < 0.3, ZP>+30 mV). pCasSA was loaded into the Lip, to form a lipoplex (Lpx), a complex of Lip and pCasSA, via electrostatic interactions. Cytotoxicity studies using mouse fibroblast L929 cells revealed that cell viabilities remained at approximately 93% and 83% following treatment with the highest concentration of Lip and Lpx formulations, respectively. Successful gene editing was confirmed via Sanger sequencing and RT-qPCR, showing a 75% reduction in ermA expression when Lpx wascombined with sonoporation. Phenotypic assays demonstrated a significant restoration of susceptibility. The minimum inhibitory concentration (MIC) of erythromycin decreased 8-fold (from 8 mg/L to 1 mg/L), and inhibition zones increased 2.5-fold (from 12 mm to 30 mm). These findings suggest that Lpx-mediated CRISPR-Cas9 delivery is a highly efficient strategy for antibiotic resensitization, potentially restoring the clinical utility of existing antimicrobials.