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The Chinese caterpillar mushroom, Ophiocordyceps sinensis (O. sinensis), is a rarely medicinal fungus in traditional chinese herbal medicine due to its unique medicinal values, and the expression stability of reference genes is essential to normalize its gene expression analysis. In this study, BestKeeper, NormFinder and geNorm, three authoritative statistical arithmetics, were applied to evaluate the expression stability of sixteen candidate reference genes (CRGs) in O. sinensis under different stress [low temperature (4°C), light treatment (300 lx), NaCl (3.8%)] and different development stages (mycelia, primordia and fruit bodies) and formation of morphologic mycelium (aeriasubstrate, hyphae knot mycelium). The paired variation values indicated that two genes could be enough to accurate standardization exposed to different conditions of O.sinensis. Among these sixteen CRGs, 18S ribosomal RNA (18S rRNA) and beta-Tubulin (β-TUB) showed the topmost expression stability in O.sinensis exposed to all conditions, while glutathione hydrolase proenzym (GGT) and Phosphoglucose isomerase (PGI) showed the least expression stability. The optimal reference gene in different conditions was various. β-TUB and Ubiquitin (UBQ) were identified as the two most stable genes in different primordia developmental stage, while phosphoglucomutase (PGM) with elongation factor 1-alpha (EF1-α) and 18S rRNA with UBQ were the most stably expressed for differentially morphologic mycelium stages and different stresses, respectively. These results will contribute to more accurate evaluation of the gene relative expression levels in O.sinensis under different conditions using the optimal reference gene in real-time quantitative PCR (RT-qPCR) analysis.

Sleep deprivation (SD) disrupts circadian rhythms; however, its effects on SD and the mechanisms involved require further investigation. Previous studies on SD were mainly conducted on rodents, such as mice, with few studies on its effects on the liver of large diurnal animals, such as sheep. In this study, we used a Tibetan sheep model for the first time to investigate the effects of SD on the liver by exposing Tibetan sheep (Ovis aries) to 7 days of SD (6 h/day) and performed transcriptome sequencing analysis on liver samples taken at 4 h intervals over 24 h. The results revealed that SD significantly altered the circadian expression of genes and their expression patterns in the liver of Tibetan sheep. Enrichment analysis of the circadian rhythm-altered genes revealed changes in the pathways related to lipid metabolism in the liver. Further evidence from serum markers and gene expression analyses using qualitative real-time polymerase chain reaction and Oil Red O and apoptosis staining indicated that SD leads to abnormal lipid metabolism in the liver, potentially causing liver damage. Therefore, our results suggest that SD disrupts the circadian rhythms of metabolism-related genes in the Tibetan sheep liver, thereby affecting metabolic homeostasis.

Simple Summary: Sleep is a vital physiological process essential for maintaining mental and physical health. Insufficient sleep affects various physiological processes, including liver function and metabolism. In this study, we used a Tibetan sheep model for the first time to investigate the effects of sleep deprivation (SD) for 7 days (6 h/day) on liver function and circadian rhythms. We found that SD significantly disrupted the expression patterns of circadian rhythm genes in the liver. Gene enrichment analysis further indicated alterations in pathways related to lipid metabolism in the liver, suggesting that SD resets the expression of metabolic genes, causing liver damage by disrupting lipid metabolism. Sleep deprivation (SD) disrupts circadian rhythms; however, its effects on SD and the mechanisms involved require further investigation. Previous studies on SD were mainly conducted on rodents, such as mice, with few studies on its effects on the liver of large diurnal animals, such as sheep. In this study, we used a Tibetan sheep model for the first time to investigate the effects of SD on the liver by exposing Tibetan sheep (Ovis aries) to 7 days of SD (6 h/day) and performed transcriptome sequencing analysis on liver samples taken at 4 h intervals over 24 h. The results revealed that SD significantly altered the circadian expression of genes and their expression patterns in the liver of Tibetan sheep. Enrichment analysis of the circadian rhythm-altered genes revealed changes in the pathways related to lipid metabolism in the liver. Further evidence from serum markers and gene expression analyses using qualitative real-time polymerase chain reaction and Oil Red O and apoptosis staining indicated that SD leads to abnormal lipid metabolism in the liver, potentially causing liver damage. Therefore, our results suggest that SD disrupts the circadian rhythms of metabolism-related genes in the Tibetan sheep liver, thereby affecting metabolic homeostasis. [ABSTRACT FROM AUTHOR]

Heat stress (HS) has become a common stressor, owing to the increasing frequency of extreme high-temperature weather triggered by global warming, which has seriously affected the reproductive capacity of important livestock such as sheep. However, little is known about whether HS reduces sperm motility by inducing circadian rhythm disorders in rumen microorganisms and metabolites in sheep. In this study, the year-round reproduction of two-year-old Hu rams was selected, and the samples were collected in May and July 2022 at average environmental temperatures between 18.71 °C and 33.58 °C, respectively. The experiment revealed that the mean temperature-humidity index was 86.34 in July, indicating that Hu rams suffered from HS. Our research revealed that HS significantly decreased sperm motility in Hu rams. Microbiome analysis further revealed that HS reshaped the composition and circadian rhythm of rumen microorganisms, leading to the circadian disruption of microorganisms that drive cortisol and testosterone synthesis. Serum indicators further confirmed that HS significantly increased the concentrations of cortisol during the daytime and decreased the testosterone concentration at the highest body temperature. Untargeted metabolomics analysis revealed that the circadian rhythm of rumen fluid metabolites in the HS group was enriched by the cortisol and steroid synthesis pathways. Moreover, HS downregulated metabolites, such as kaempferol and L-tryptophan in rumen fluid and seminal plasma, which are associated with promotion of spermatogenesis and sperm motility; furthermore, these metabolites were found to be strongly positively correlated with Veillonellaceae_UCG_001. Overall, this study revealed the relationship between the HS-induced circadian rhythm disruption of rumen microorganisms and metabolites and sperm motility decline. Our findings provide a new perspective for further interventions in enhancing sheep sperm motility with regard to the circadian time scale. [ABSTRACT FROM AUTHOR]

Simple Summary: This study compared the growth and meat quality of juvenile male and female Nile tilapia. Males grew bigger and heavier, while females excelled in feed conversion. Males had more crude fat and ash, but moisture and protein levels were unchanged. Amino acid profiles were similar overall, though females had an advantage in non-essential amino acids. This study compared the growth and flesh quality of juvenile male and female Nile tilapia grown in separate RAS tanks. The Genetic Sex Determination method yielded 40 males and 40 females. Males grew longer and heavier than females, and the results showed significant variation between the sexes in terms of weight gain rate (WGR), specific growth rate (SGR), and final body length and weight. In terms of the feed conversion ratio (FCR), Gonad Somatic Index (GSI), and Viscera Somatic Index (VSI), females presented better results than males. Male and female Hepatosomatic Index (HSI) values did not differ significantly—no difference in blood serum values. The meat's moisture and crude protein composition did not alter significantly from male to female; however, the crude fat and ash levels did differ significantly. Male and female animals were given the same seventeen distinct types of amino acids, and there was no distinct variation in the profiles of total amino acids (TAAs) and essential amino acids (EAAs) between the sexes. However, in the non-essential amino acid (NEAA) category, there were marginally significant differences, wherein females performed better than males. Males and females differed considerably in crude fat and ash levels but not in the moisture content or crude protein composition of the meat. Regarding fatty acids, males outperformed females in terms of total fatty acids (TFAs), polyunsaturated fatty acids (PUFAs), and saturated fatty acids (SFAs). However, no significant difference in the amount of monounsaturated fatty acids (MUFAs) in muscle was found between males and females. [ABSTRACT FROM AUTHOR]

Developing high‐performance lithium‐sulfur batteries is a promising way to attain higher energy density at a lower cost beyond the state‐of‐the‐art lithium‐ion battery technology. However, the major issues impeding their practical applications are the sluggish kinetics and the parasitic shuttling reactions of sulfur and polysulfides. Here, pillaring the multilayer graphene membrane with a metal–organic framework (MOF) demonstrates the substantial impact of a versatile interlayer design in tackling these issues. Unlike regular composite separators reported so far, the participation of tri‐metallic Ni−Co−Mn MOF as pillars supports the construction of an ion‐channel interconnected interlayer structure, unexpectedly balancing the interfacial concentration polarization, spatially confining the soluble polysulfides, and vastly affording the lithiophilic sites for highly efficient polysulfide sieving/conversion. As a demonstration, we show that the MOF‐pillared interlayer structure enables outstanding capacity (1634 mAh g−1 at 0.1 C) and longevity (average capacity decay of 0.034 % per cycle in 2000 cycles) for lithium‐sulfur batteries. Besides, the multilayer separator can be readily integrated into the high‐nickel cathode (LiNi0.91Mn0.03Co0.06O2)‐based lithium‐ion batteries, which efficiently suppresses the undesired phase evolution upon cycling. These findings suggest the potential of “gap‐filling” materials in fabricating multi‐functional separators, bringing forward the pillared interlayer structure for energy‐storage applications. [ABSTRACT FROM AUTHOR]

Simple Summary: Capacitation confers competency to spermatozoa to fertilize the oocyte, yet its regulatory mechanisms are not fully understood. Thus, we aimed to investigate the comparative proteomic profiling in ram spermatozoa under non-capacitating (NC) and capacitating (CAP) conditions in vitro using a liquid chromatography–tandem mass spectrometry (LCMS/MS) combined with tandem mass tag (TMT) labeling strategy. Functional enrichment analysis indicated that the differentially abundant proteins Prune Exopolyphosphatase 1, Galactose-1-Phosphate Uridylyltransferase, and ATP Citrate Lyase were strictly related to energy production and conversion, and Phosphoglycolate phosphatase, Glucosamine-6-Phosphate Deaminase 1 and 2 were related to metabolism, RNA processing, and vesicular transport pathways. Furthermore, the networks of protein–protein interaction indicated a strong interaction among these differential proteins in annotated pathways such as ubiquitin and transport metabolism. Together, our results provided the database for studying the differentially expressed proteins during ram sperm capacitation. Ram sperm undergo a sequence of physiological and biochemical changes collectively termed as capacitation to perform oocyte fertilization. However, the protein changes induced by capacitation remain in need of further exploration. Thus, the present study investigated the comparative proteomic profiling in ram spermatozoa under non-capacitating (NC) and capacitating (CAP) conditions in vitro using a liquid chromatography–tandem mass spectrometry combined with tandem mass tag labeling strategy. As a results, 2050 proteins were identified and quantified; 348 of them were differentially abundant, with 280 of the proteins upregulated and 68 of the proteins downregulated between the CAP and NC spermatozoa, respectively. Functional enrichment analysis indicated that the differentially abundant proteins Prune Exopolyphosphatase 1, Galactose-1-Phosphate Uridylyltransferase, and ATP Citrate Lyase were strictly related to energy production and conversion, and Phosphoglycolate phosphatase, Glucosamine-6-Phosphate Deaminase 1 and 2 were related to metabolism, RNA processing, and vesicular transport pathways. Furthermore, the networks of protein–protein interaction indicated a strong interaction among these differential proteins in annotated pathways such as ubiquitin and transport metabolism. Our findings indicate that capacitation progress might be regulated through different pathways, providing insights into mechanisms involved in ram sperm capacitation and fertility. [ABSTRACT FROM AUTHOR]

The high-precision and high-purity isolation of circulating tumor cells (CTCs) from whole blood is vital to early cancer detection. Cascaded microfluidic separation is highly efficient because it connects multiple-stage separations in series. Here, we numerically investigated sheathless tumor cell separation with size-dependent cascaded inertial and deterministic lateral displacement (DLD) microfluidic device. The inertial microfluidic is arranged in the first-stage unit for particle focusing and rough sorting, and the cascaded DLD microfluidic is arranged in the second stage for realizing further sorting and purification. A parametric study with flow rate range from 100–600 μl/min and aspect ratio range from 60:100 to 60:300 of the first stage was carried out to optimize channel structure for realizing high-efficiency separation. Then, the pre-separation mechanism within the spiral microchannel was analyzed. The purity of the obtained CTCs and the separation efficiency were further improved using a droplet-type microcolumn DLD microfluidic device as the second unit. The cascade eliminates the need for additional force fields and reduces device complexity while simplifying operation and reducing the chance of sample contamination. [ABSTRACT FROM AUTHOR]