Search Results (2,068)

MicroRNAs (miRNAs) are short, non-coding RNA molecules that play a crucial role in regulating various biological processes by influencing post-transcriptional gene expression and gene silencing. Background/Objectives: In this study, rabbit embryos were utilised as a model system to investigate potential biomarkers relevant to human embryo development. Seven microRNAs (miRNAs) identified in the embryo culture medium were evaluated as biomarkers by analysing the correlation between their expression levels and the developmental quality of rabbit embryos at days 4 and 6. Methods: We analysed the expression of seven development-specific miRNAs (miR-24-3p, miR-28-3p, miR-103a-3p, miR-181a-5p, miR-191-5p, miR-320a-3p, miR-378a-3p) in 4-day-old and 6-day-old rabbit embryos, along with their culture media. Results: Our findings revealed significant differences in the expression levels of these miRNAs between the 4-day-old and 6-day-old embryos. On the other hand, the expression patterns observed in the culture medium samples showed less variation between the two age groups. Nonetheless, analysis of miRNA expression profiles in the spent culture medium from individually cultured embryos enabled the identification of lower-quality embryos, characterised by smaller size and impaired or delayed development. Conclusions: The detection of these miRNAs in embryo culture medium may serve as a reliable indicator of successful progression to the blastocyst stage. Our experimental results identified specific miRNAs whose expression profiles differ according to embryonic stage and quality, thereby reflecting key developmental milestones. Notably, the detectability of these miRNAs in the medium—without prior RNA isolation—indicates their active secretion into the extracellular environment. By synthesising our findings with the existing literature, we refined a panel of miRNAs essential for the development of implantation-competent embryos in both rabbits and humans. Consequently, we developed a non-invasive assay for predicting implantation and pregnancy outcomes, which may have significant applications in human reproductive medicine. Full article

Lung development is governed by tightly regulated signaling mechanisms, including endocytosis-mediated pathways critical for epithelial–mesenchymal communication and tissue remodeling. This study investigated the effects of Dab1 deficiency on the expression of endocytic and signaling-related proteins, Megalin, Cubilin, Caveolin-1, GIPC1, and Dab2IP, during embryonic lung development in yotari mice. Using immunofluorescence and quantitative image analysis, protein expressions were compared between yotari and wild-type embryos at gestational days E13.5 and E15.5. Results showed significantly reduced expression of Caveolin-1 in the yotari epithelium across both stages, along with diminished mesenchymal levels of Megalin and GIPC1 at E13.5. Cubilin and Dab2IP expression patterns showed no statistically significant differences, although developmental and compartmental shifts were observed. These findings suggest that Dab1 deficiency selectively disrupts endocytic and signaling scaffolds crucial for branching morphogenesis and alveolar maturation. The altered spatiotemporal expression of these proteins underscores the essential role of Dab1 in regulating lung epithelial–mesenchymal dynamics and maintaining developmental homeostasis during critical stages of organogenesis. Full article

Plodia interpunctella is a globally significant pest of stored grains, posing a major threat to food safety. To explore its cold-adaptation mechanisms, this study evaluated the physiological and developmental responses of different life stages following short-term cold acclimation at 4 °C. Results showed that cold acclimation significantly reduced the supercooling points (SCPs) of larvae and pupae, with the greatest reduction observed in the second instar larvae. Antioxidant enzyme assays revealed marked increases in the activities of superoxide dismutase (SOD), catalase (CAT), and peroxidase (POD), indicating enhanced oxidative stress resistance. Developmental durations were significantly shortened at lower temperatures in acclimated individuals, and fecundity was notably increased at 24 °C, although no significant changes were observed at higher temperatures. These findings suggest that cold acclimation improves the cold tolerance and reproductive performance of P. interpunctella under low-temperature conditions, offering insights into insect adaptability and providing theoretical support for the development of low-temperature-based pest management strategies in stored grain systems. Full article

Trypanosoma cruzi is a hemoflagellate protozoan and the causative agent of Chagas disease, also known as American trypanosomiasis. Transmission occurs through the feces of triatomine insects, its biological vector. It is estimated that around 7 million people are infected across Mexico, Central America, and South America. This study aimed to identify and characterize T. cruzi isolates obtained from wild triatomine vectors collected in Aguascalientes, Mexico. Molecular identification was performed at different developmental stages—epimastigotes in culture media, metacyclic trypomastigotes in triatomine feces, and amastigotes in mouse cardiac tissue—using endpoint PCR targeting satDNA and mtCytB regions. In addition, next-generation sequencing was employed to analyze variable regions of kinetoplast DNA minicircles. The pathogenicity of the isolated and identified T. cruzi strain was assessed in a murine model, where trypomastigote stages were detected in peripheral blood and amastigote stages in muscle tissue. Molecular analyses confirmed the presence of T. cruzi across different developmental stages from wild vectors, demonstrating that the isolated wild strain possesses pathogenic potential when completing its life cycle in an experimental mammalian host, specifically BALB/c mice. Full article

Although traditional genetically modified (GM) cotton has reduced lepidopteran pests, secondary pests such as Adelphocoris suturalis Jakovlev (Hemiptera: Miridae) have become increasingly problematic. RNA interference (RNAi)-based insect-resistant plants offer a promising alternative, but their potential ecological risks, particularly within the plant–pest–natural enemy framework, require comprehensive evaluation. As a key natural enemy in cotton agroecosystems, Harmonia axyridis Pallas (Coleoptera: Coccinellidae) plays a vital role in regulating pest populations and thus serves as an important non-target insect for environmental risk assessment. In this study, dsAsFAR transgenic cotton and the non-target insect H. axyridis were used to evaluate potential ecological risks. Based on the sequences of the HaFAR, AsFAR, and GFP genes, dsAsFAR, dsHaFAR, and dsGFP were synthesized in vitro and incorporated into artificial diets fed to H. axyridis, along with a ddH₂O control group. No significant differences were observed among the treatments across various developmental stages. Furthermore, H. axyridis was fed aphids that had been reared on either transgenic or non-transgenic cotton plants cultivated in a greenhouse. The findings indicated no significant differences in the growth, development, predatory ability, or fecundity of H. axyridis. These results suggest that transgenic dsAsFAR cotton targeting A. suturalis poses no detectable adverse effects on H. axyridis, supporting the environmental safety of RNAi crops within a plant–pest–natural enemy ecological interaction framework. Moreover, the dsAsFAR expressed in transgenic cotton was not effectively transferred through the food chain to affect homologous gene expression in H. axyridis. Full article

The BBX gene family functions as a key transcription factor implicated in plant growth, development, and stress responses. However, research on this gene family in melon remains absent. In the present study, we identified 19 BBX family genes within the melon genome, distributed across chromosomes 1, 2, 3, 4, 5, 7, 8, 10, 11, and 12. Phylogenetic analysis categorized these genes into five distinct subfamilies, with notable similarities observed in gene structure and conserved motifs among members of the same subfamily. Synteny analysis revealed seven syntenic relationships among melon BBX genes, 17 between melon and Arabidopsis, and one between melon and rice. Reanalysis of transcriptome data indicated that certain BBX genes exhibit high expression levels across various tissues and developmental stages of fruits, while others display tissue specificity. Under both abiotic and biotic stress conditions, genes such as CmBBX3, CmBBX5, CmBBX2, CmBBX18, CmBBX15, and CmBBX11 demonstrated significant differential expression, highlighting their critical roles in melon growth and development. Additionally, RT-qPCR analysis was conducted to examine the expression levels of melon BBX genes at different time points under salt stress, further validating the transcriptome data. This study provides a theoretical foundation for future molecular breeding efforts in melon. Full article

Propsilocerus akamusi (Tokunaga, 1938) larvae serve as key bioindicators for water quality assessment. This study identifies optimal reference genes for RT-qPCR under diverse experimental conditions. Fifteen candidate genes commonly employed in other insect species were selected, candidate genes commonly used in other insect species. Homologous genes were identified in the P. akamusi genome through sequence alignment with their Drosophila melanogaster counterparts. Expression stability across developmental stages, body parts, temperature variations, and deltamethrin/nickel chloride exposures was systematically evaluated using geNorm, NormFinder, BestKeeper, and ΔCt methods. RPL32 exhibited the highest expression stability across different body parts of adults under varying temperature conditions, and RPS11 and RPL8 showed the greatest stability across developmental stages and in larvae exposed to different temperatures. Furthermore, under nickel chloride and deltamethrin treatments, RPS11 and RPL8 maintained the highest expression stability. The results indicated that the expression stability of reference genes varied under different conditions. Among different body parts of adults, RPL32 and RPL4 exhibited the most stable expression. Across different developmental stages, RPS11 and RPL8 performed best in terms of expression stability. Under different temperature treatments, RPL32 and RPL4 remained stable in adults, while RPS11 and RPL8 showed the greatest stability in larvae. Similarly, under nickel chloride and deltamethrin treatments, RPS11 and RPL8 demonstrated the most stable expression in larvae. Through the above research, we can advance ecosystem-impact insights and bolster environmental protection and water quality monitoring. Full article

Pepper is a major horticultural crop cultivated extensively worldwide. Among its various agronomic characteristics, fruit length is a key trait influencing both yield and visual quality. Despite its importance, the genetic mechanisms regulating fruit length in Capsicum remain insufficiently characterized, hindering the development of high-yielding and aesthetically desirable cultivars. In this study, fruits at three developmental stages (0, 15, and 30 days after flowering) were sampled from the long-fruit mutant fe1 and its wild-type progenitor LY0. Phenotypic characterization and transcriptomic sequencing were conducted to identify candidate genes associated with fruit length regulation. Morphological analysis revealed that the most pronounced difference in fruit length occurred at 30 days after flowering. RNA-seq analysis identified 41,194 genes, including 13,512 differentially expressed genes (DEGs). Enrichment analysis highlighted key pathways, such as plant–pathogen interaction, plant hormone signal transduction, and the MAPK signaling pathway. DEG classification suggested that several downregulated genes related to early auxin responses may contribute to the regulation of fruit elongation. Notably, the gibberellin signaling gene SCL13 (Caz12g26660), transcription factors MYB48 (Caz11g07190) and ERF3-like (Caz10g00810), and the cell-wall-modifying gene XTH15-like (Caz07g19100) showed significantly elevated expression in 30-day-old fruits of fe1. Weighted gene co-expression network analysis (WGCNA) further revealed a strong positive correlation among these genes. Quantitative RT-PCR analysis of eight selected DEGs confirmed the RNA-seq results. This study provides a foundational framework for dissecting the molecular regulatory network of fruit length in Capsicum, offering valuable insights for breeding programs. Full article

Antheraea pernyi (Lepidoptera: Saturniidae) is an economically important silk-producing insect, whose gut microbiota play a crucial role in growth, development, and nutrient metabolism. This study focused on the entire larval developmental stages of A. pernyi. Using the Illumina MiSeq high-throughput sequencing platform, we performed 16S rRNA gene amplicon sequencing on the gut microbiota of laboratory-reared A. pernyi larvae, analyzing in detail the composition and diversity characteristics of the gut microbial communities across all five instars (1st to 5th instar). Additionally, functional predictions were conducted to explore the potential roles of these microbiota during larvae development. The study revealed that the core gut microbiota of A. pernyi larvae primarily consisted of Actinomycetota (39.78%), Cyanobacteriota (32.46%), Bacillota (18.08%), and Pseudomonadota (9.02%). Among these, Actinomycetota dominated in the 1st to 4th-instar larvae, while Cyanobacteriota became the predominant phylum in the 5th instar. Linear discriminant analysis effect size identified statistically significant biomarkers across different instar larvae of A. pernyi. Alpha diversity analysis showed that gut microbiota diversity initially increased and then decreased with larval development, peaking in the 3rd instar, and reaching its lowest level in the 5th instar. Principal coordinate analysis (PCoA) of beta diversity indicated that the gut microbiota structures of the 1st to 4th instars were similar but significantly differed from that of the 5th instar. Functional prediction analysis based on the KEGG database revealed that Carbohydrate metabolism and Amino acid metabolism-related genes were significantly lower in the 5th instar compared to other instars, while Energy metabolism and Cofactor and vitamin metabolism-related genes were significantly higher. This study offers valuable insights for the development of gut microbial resources in Lepidoptera insects. Full article

Aspergillus fumigatus is an opportunistic filamentous fungus that primarily affects the respiratory tract of the human body. Depending on its host’s immune response, the pathogen can cause invasive pulmonary aspergillosis (IPA). Biofilm formation by A. fumigatus increases virulence and resistance against antifungals and immune response and is one important factor in IPA development. Here, two human-like models, precision cut lung slices (PCLS) and a biofilm co-culture model, have been developed to test the anti-biofilm activity of voriconazole, amphotericin B, as well as luliconazole against A. fumigatus. In both assays, metabolically active A. fumigatus biofilms were examined at different biofilm developmental stages using an XTT assay. A decrease in the metabolic activity of the fungal biofilms was detected for each of the tested agents in both assays. Significant anti-biofilm effects exist against early-stage biofilm in the co-culture model. In the PCLS assay, amphotericin B showed the strongest inhibition after 24 h. In conclusion, the applied PCLS ex vivo model can be used to study the property and activity of certain antifungal compounds against Aspergillus biofilm. With its close resemblance to human conditions, the PCLS model has the potential for improving the current understanding of biofilm treatments in laboratory settings. Full article

Starch synthase collaboratively accomplishes the synthesis of starch by forming a protein complex. In barley (Hordeum vulgare), the interaction mechanisms within the soluble starch synthase 3 (SSIII) protein complex and their specific roles in starch biosynthesis remain poorly understood. To investigate proteins interacting with SSIII isoforms (SSIIIa/SSIIIb), the polyclonal antibodies against HvSSIIIa and HvSSIIIb were generated in rabbits. Co-immunoprecipitation (Co-IP) was employed to enrich HvSSIIIa- and HvSSIIIb-associated protein complexes from developing barley seeds. The potential interacting proteins were identified using mass spectrometry coupled with database searches. The results demonstrated the successful production of highly specific polyclonal antibodies against HvSSIIIa and HvSSIIIb. These antibodies were used to analyze the expression levels of HvSSIIIa and HvSSIIIb across different barley tissue and seed developmental stages. Both isoforms exhibited peak expression during early seed development. Further Co-IP-based screening revealed several candidate interacting proteins, and the interacting proteins had some overlaps. Among them, adenosine diphosphate glucose pyrophosphorylase, as a key rate-limiting enzyme in starch synthesis, interacts with SSIII to jointly regulate the supply of ADP-glucose precursors. Differential proteins show functional differentiation. In SSIIIa, the interacting protein DELLA protein SLR1-like is involved in processes such as hormone regulation, and in SSIIIb, the interacting protein 14-3-3 is related to metabolism. These findings provide new insights into the composition and function of the SSIII protein complex in barley, facilitating future studies on its regulatory role in starch biosynthesis. Full article

This study evaluated the effects of red and blue laser irradiation on the development and reproduction of the Asian corn borer (Ostrinia furnacalis (Guenée)) under controlled laboratory conditions, aiming to explore its potential for non-chemical pest control. Larvae were exposed to laser light at different wavelengths and intensities, and key biological parameters—including egg hatching, larval duration, pupation, adult emergence, and oviposition—were assessed. Red laser light slightly delayed egg hatching but had minimal effects on subsequent developmental stages. In contrast, blue laser irradiation significantly prolonged the larval period and reduced pupation rates. Combined red–blue treatments produced similar inhibitory effects to blue light alone, suggesting that blue wavelengths were the primary factor driving developmental delays. These findings demonstrate that blue and red–blue laser irradiation can effectively interfere with the life cycle of O. furnacalis, offering a promising approach for sustainable, light-based pest management strategies. Full article

Background: Problematic Internet gaming (PIG), considered an early stage of Internet gaming addiction (IGA), has become increasingly prevalent among adolescents. This study focused on deviant peer affiliation (DPA) and hedonic gaming experience (HGE) as key mediators and examined four psychosocial risk factors closely related to them: interpersonal incompetence (II), perceived stress (PS), frustration (FR), and emotional loneliness (EL). Specifically, the study investigated how these four psychosocial risk factors influence adolescents’ DPA, HGE, and PIG, and whether DPA and HGE mediate these relationships. Methods: Based on existing validated scales, we developed a questionnaire to measure these seven constructs (II, PS, FR, IC, DPA, HGE, and PIG), proposed 14 hypotheses, and collected 214 valid responses from adolescents. Structural equation modeling (SEM) was used to test the hypothesized model. Findings: The results showed that all 14 hypotheses were supported. Specifically, interpersonal incompetence significantly predicted perceived stress; stress led to frustration; and frustration, in turn, contributed to emotional loneliness. Furthermore, all four psychosocial risk factors significantly predicted deviant peer affiliation, hedonic gaming experience, and ultimately, problematic Internet gaming among adolescents. Both DPA and HGE mediated the effects of psychosocial risk factors on adolescent problematic Internet gaming (PIG), with the model explaining moderate-to-high variance. This study highlights the importance of segmenting adolescents into more specific subgroups based on the distinct developmental pathways leading to PIG. Implications: Understanding the step-by-step mechanisms and psychological drivers of different adolescent subtypes can provide a more solid foundation for early identification and targeted intervention efforts. Full article

Forests, both natural and managed, provide a critical habitat for a significant part of global biodiversity. Among many different groups of forest biota, tree species occupy a special position as they create conditions upon which the existence of virtually all other forest organisms depends, either directly or indirectly. To permanently play this role, particular tree species must be demographically stable; i.e., their populations should be distinguished by the balanced, size-dependent proportions of individuals representing different developmental stages (from seedlings and saplings to mature and old trees). In this study, we examined the extent to which this condition is met in the managed part of Białowieża Forest in northeastern Poland, an important biodiversity hotspot in Central Europe. Comparison of species-specific equilibrium vs. actual size distributions revealed that almost half of all trees growing in Białowieża Forest represented “inappropriate” (i.e., occurring in excess compared to the balanced models) species and/or diameter ranges. The amount of deficits was also large (around 30% of the current tree number), concerning primarily the smallest trees. Considering this, we recommend targeted, active management strategies to restore the demographic balance of key tree species and, thus, to enhance the conservation of local biodiversity. We also indicate that the key elements of such strategies should be the gradual removal of trees from surplus diameter ranges and assisted regeneration of species with the greatest deficiencies in small diameter classes. Full article

Intra-specific variation in functional traits and their inter-relationships reflect how plants allocate resources, adapt, and evolve in response to environmental changes. This study investigated eight functional traits—leaf area (LA), specific leaf area (SLA), leaf dry matter content (LDMC), chlorophyll content (CHL), leaf nitrogen content (LNC), leaf phosphorus content (LPC), twig tissue density (TTD), and wood density (WD)—in Cunninghamia lanceolata plantations of three stand ages (15, 30, and 50 years), using a space-for-time substitution approach. We examined differences in trait values, intra-specific variation, and trait correlations across forest ages and diameter classes. The results showed that (1) Functional traits exhibited varying degrees of intra-specific variation, with LA having the highest coefficient of variation (21.66%) and LPC is lowest (9.31%). (2) Forest age had a stronger influence on trait variation than diameter class, with all traits differing significantly across ages, while only WD varied significantly among diameter classes. (3) PC1 (25.5%) and PC2 (19.4%) together explained approximately 44.9% of the total variation, with PC1 primarily reflecting functional trait changes driven by forest age. PCA results showed that LA and CHL tended to exhibit higher values in young forests, whereas SLA, LDMC, LPC, and LNC had relatively higher values in mature forests. This pattern suggests a shift in functional trait expression from resource acquisition to resource conservation strategies with increasing forest age. (4) Significant positive correlations between LNC and LPC, and negative correlations between SLA and LDMC, were observed in most groups, except in large-diameter trees at the over-mature stage. C. lanceolata adjusts trait combinations to enhance fitness across developmental stages. Juvenile trees adopt traits favoring efficient light and nutrient use to support rapid growth and competition. Middle-aged trees prioritize balanced water and nutrient use to maintain productivity and resist disturbances. Mature trees focus on sustained resource use and offspring protection to support ecosystem stability and regeneration. These findings reveal age-specific adaptive strategies and provide insights into the coordination and trade-offs among traits in response to environmental conditions. Full article