Search Results (208)

Bud eye identification is a critical step in the intelligent seed cutting process for potatoes. This study focuses on the challenges of low testing accuracy and excessive weighted memory in testing models for potato bud eye detection. It proposes an improved potato bud eye detection method based on YOLOv5s, referred to as the YOLO-SCA model, which synergistically optimizing three main modules. The improved model introduces the ShuffleNetV2 module to reconstruct the backbone network. The channel shuffling mechanism reduces the model’s weighted memory and computational load, while enhancing bud eye features. Additionally, the CBAM attention mechanism is embedded at specific layers, using dual-path feature weighting (channel and spatial) to enhance sensitivity to key bud eye features in complex contexts. Then, the Alpha-IoU function is used to replace the CloU function as the bounding box regression loss function. Its single-parameter control mechanism and adaptive gradient amplification characteristics significantly improve the accuracy of bud eye positioning and strengthen the model’s anti-interference ability. Finally, we conduct pruning based on the channel evaluation after sparse training, accurately removing redundant channels, significantly reducing the amount of computation and weighted memory, and achieving real-time performance of the model. This study aims to address how potato bud eye detection models can achieve high-precision real-time detection under the conditions of limited computational resources and storage space. The improved YOLO-SCA model has a size of 3.6 MB, which is 35.3% of the original model; the number of parameters is 1.7 M, which is 25% of the original model; and the average accuracy rate is 95.3%, which is a 12.5% improvement over the original model. This study provides theoretical support for the development of potato bud eye recognition technology and intelligent cutting equipment. Full article

Double-petal ornamental pomegranate presents for its enhanced ornamental value. Thus, cultivation techniques that promote petaloidy while modulating related gene expression are desired. To screen out the efficient treatments of plant growth regulator and key genes that enhance petaloidy, this study treated the flower buds of double- and single-petal ornamental pomegranate varieties with different concentrations of plant growth regulators naphthaleneacetic acid (NAA), methyl jasmonate (MeJA), abscisic acid (ABA), and ethephon (ETH) and quantified the number of petalized stamens (NOPSs) and the number of petals (NOPs) in both varieties. Furthermore, we investigated the expression levels of the genes flavin-containing monooxygenase (YUC), IAA-amino acid hydrolase (ILR1),indole-3-acetic acid-amido synthetase (GH3.17), auxin transporter (LAX2), auxin response factor (ARF), auxin-induced in root cultures protein (AIR12), jasmonic acid-amido synthetase (JAR1), and ABA stress ripening-induced protein (ASR) under the different treatments and analyzed their role in regulating relevant phenotypic traits. Plant growth regulator experiments demonstrated that NAA (10 mg/L) significantly increased the number of petalized stamens (NOPSs) and petals (NOPs), MeJA (100 mg/L) significantly increased the number of petalized stamens, while neither ABA nor ETH induced this morphological shift. qRT-PCR analysis confirmed that NAA upregulated ILR1, LAX2, ARF, and JAR1 in the stamens of single-petal flowers (StSi) and double-petal flowers (StDo) and petals of single-petal flowers (PeSi) and double-petal flowers (PeDo), with their expression levels strongly positively correlated with NOPS in both single- and double-petal flowers and NOP in double-petal flowers. MeJA upregulated ILR1, GH3.17, LAX2, ARF, and JAR1 in StDo and PeDo and was strongly positively correlated with NOPS and NOP in double-petal flowers. Consequently, NAA (10 mg/L) and MeJA (100 mg/L) were efficient treatments, and ILR1, GH3.17, LAX2, ARF, and JAR1 were identified as key genes in NAA- and MeJA-mediated petaloidy in ornamental pomegranates. Our results provide theoretical support for identifying the formation mechanism and improving industrial cultivation techniques for double-petal pomegranates. Full article

Crataegus harbisonii Beadle (Harbison’s or Harbison hawthorn) is a Tennessee (USA) endemic tree of the Rosaceae family, currently considered “critically imperiled” at the state, national, and global levels. It is known from only two extant wild locations, one in Davidson County, Tennessee consisting of a single living individual and a population of less than 100 individuals in Obion County, Tennessee. Key ex situ conservation efforts undertaken over the last three years with this critically imperiled species are reported here. The Obion County population was intensively surveyed and all C. harbisonii individuals documented. Over three seasons, seeds were collected and propagated, and clones were generated via chip-budding and grafting. Conservation seed orchards were planned and established to provide a stable, long-term source of genetically robust seed for reforestation and research. To date, 19 sources from the Obion County location as well as the single Davidson County genotype have been successfully preserved through clonal propagation, and open-pollinated seedlings produced from 12 unique mother trees. Additional material is being added annually. We report lessons learned as well as key future research directions, now enabled through the establishment of germplasm resources. Full article

Background/Objectives: Smart Formulation is an artificial intelligence-based platform designed to predict the Beyond Use Dates (BUDs) of compounded oral solid dosage forms. The study aims to develop a decision-support tool for pharmacists by integrating molecular, formulation, and environmental parameters to assist in optimizing the stability of extemporaneous preparations. Methods: A tree ensemble regression model was trained using a curated dataset of 55 experimental BUD values collected from the Stabilis database. Each formulation was encoded with molecular descriptors, excipient composition, packaging type, and storage conditions. The model was implemented using the KNIME platform, allowing the integration of cheminformatics and machine learning workflows. After training, the model was used to predict BUDs for 3166 APIs under various formulation and storage scenarios. Results: The analysis revealed a significant impact of excipient type, number, and environmental conditions on API stability. APIs with lower LogP values generally exhibited greater stability, particularly when formulated with a single excipient. Excipients such as cellulose, silica, sucrose, and mannitol were associated with improved stability, whereas HPMC and lactose contributed to faster degradation. The use of two excipients instead of one frequently resulted in reduced BUDs, possibly due to moisture redistribution or phase separation effects. Conclusions: Smart Formulation represents a valuable contribution to computational pharmaceutics, bridging theoretical formulation design with practical compounding needs. The platform offers a scalable, cost-effective alternative to traditional stability testing and is already available for use by healthcare professionals. Its implementation in hospital and community pharmacies may help mitigate drug shortages, support formulation standardization, and improve patient care. Future developments will focus on real-time stability monitoring and adaptive learning for enhanced precision. Full article

Understanding how leaf maturity affects the flavor attributes of green tea is crucial for optimizing harvest timing and processing strategies. This study comprehensively characterized the flavor profiles of Fudingdabai green teas at three distinct leaf maturity stages—single bud (FDQSG), one bud + one leaf (FDMJ1G), and one bud + two leaves (FDTC2G)—using a multimodal approach integrating electronic nose, electronic tongue, HS-GC-IMS, relative odor activity value (rOAV) evaluation, and machine learning algorithms. A total of 85 volatile compounds (VOCs) were identified, of which 41 had rOAV > 1. Notably, 2-methylbutanal, 2-ethyl-3,5-dimethylpyrazine, and linalool exhibited extremely high rOAVs (>1000). FDQSG was enriched with LOX (lipoxygenase)-derived fresh, grassy volatiles such as (Z)-3-hexen-1-ol and nonanal. FDMJ1G showed a pronounced accumulation of floral and fruity compounds, especially linalool (rOAV: 7400), while FDTC2G featured Maillard- and phenylalanine-derived volatiles like benzene acetaldehyde and 2,5-dimethylfuran, contributing to roasted and cocoa-like aromas. KEGG (Kyoto Encyclopedia of Genes and Genomes) analysis revealed significant enrichment in butanoate metabolism and monoterpenoid biosynthesis. Random forest–SHAP analysis identified 20 key flavor markers, mostly VOCs, that effectively discriminated samples by tenderness grade. ROC–AUC validation further confirmed their diagnostic performance (accuracy ≥ 0.8). These findings provide a scientific basis for flavor-driven harvest management and the quality-oriented grading of Fudingdaibai green tea. Full article

This investigation examines the function of the mouse Mycn gene in regulating and activating quiescent mammary stem cells, which are vital for mammary gland development. The mammary gland, consisting of luminal and basal cells, progresses through complex developmental stages from embryonic development through puberty, adulthood, pregnancy, lactation, and involution. Quiescent stem cells, existing in a reversible non-proliferative state, are essential for gland maintenance, yet their activation mechanisms remain poorly understood. Mycn, a member of the Myc/MYC oncogene family, is recognized for its roles in embryonic development and cancer, notably aggressive neuroblastoma and triple-negative breast cancer. Through single-cell RNA sequencing (scRNA-seq), CRISPR knockout, and overexpression experiments, this study demonstrates that Mycn is highly enriched in the terminal end buds (TEBs) of the pubertal mammary gland, particularly in basal cells, and is critical for ductal development. Both deletion and overexpression of Mycn diminish the stemness and regenerative capacity of mammary stem cells. Mycn enhances cell proliferation while downregulating quiescent stem cell markers and regulators, including Bcl11b and Tspan8, affecting stem cell maintenance and differentiation. This research clarifies the regulatory role of Bcl11b in controlling Tspan8 expression and demonstrates that Mycn indirectly targets both under normal conditions. Maintaining appropriate levels of Mycn expression is essential for normal development and cancer prevention. These insights contribute to understanding diseases and aggressive cancers, including triple-negative breast cancer (TNBC), and suggest potential therapeutic approaches. Full article

The Golgi of goblet cells represents a specialized machine for mucin glycosylation. This process occurs in a specialized form of the secretory pathway, which remains poorly examined. Here, using high-resolution three-dimensional electron microscopy (EM), EM tomography, serial block face scanning EM (SBF-SEM) and immune EM we analyzed the secretory pathway in goblet cells and revealed that COPII-coated buds on the endoplasmic reticulum (ER) are extremely rare. The ERES vesicles with dimensions typical for the COPII-dependent vesicles were not found. The Golgi is formed by a single cisterna organized in a spiral with characteristics of the cycloid surface. This ribbon has a shape of a cup with irregular perforations. The Golgi cup is filled with secretory granules (SGs) containing glycosylated mucins. Their diameter is close to 1 µm. The cup is connected with ER exit sites (ERESs) with temporal bead-like connections, which are observed mostly near the craters observed at the externally located cis surface of the cup. The craters represent conus-like cavities formed by aligned holes of gradually decreasing diameters through the first three Golgi cisternae. These craters are localized directly opposite the ERES. Clusters of the 52 nm vesicles are visible between Golgi cisternae and between SGs. The accumulation of mucin, started in the fourth cisternal layer, induces distensions of the cisternal lumen. The thickness of these distensions gradually increases in size through the next cisternal layers. The spherical distensions are observed at the edges of the Golgi cup, where they fuse with SGs and detach from the cisternae. After the fusion of SGs located just below the apical plasma membrane (APM) with APM, mucus is secreted. The content of this SG becomes less osmiophilic and the excessive surface area of the APM is formed. This membrane is eliminated through the detachment of bubbles filled with another SG and surrounded with a double membrane or by collapse of the empty SG and transformation of the double membrane lacking a visible lumen into multilayered organelles, which move to the cell basis and are secreted into the intercellular space where the processes of dendritic cells are localized. These data are evaluated from the point of view of existing models of intracellular transport. Full article

The CRISPR/Cas9 system is a powerful genome-editing tool that is applied in baculovirus engineering. In this study, we present the first report of the AcMNPV genome deletions for bioproduction purposes, using a dual single-guide RNA (sgRNA) CRISPR/Cas9 approach. We used this method to remove nonessential genes for the budded virus and boost recombinant protein yields when applied as BEVS. We show that the co-delivery of two distinct ribonucleoprotein (RNP) complexes, each assembled with a sgRNA and Cas9, into Sf9 insect cells efficiently generated deletions of fragments containing tandem genes in the genome. To evaluate the potential of this method, we assessed the expression of two model proteins, eGFP and HRPc, in insect cells and larvae. The gene deletions had diverse effects on protein expression: some significantly enhanced it while others reduced production. These results indicate that, although the targeted genes are nonessential, their removal can differentially affect recombinant protein yields depending on the host. Notably, HRPC expression increased up to 3.1-fold in Spodoptera frugiperda larvae. These findings validate an effective strategy for developing minimized baculovirus genomes and demonstrate that dual-guide CRISPR/Cas9 editing is a rapid and precise tool for baculovirus genome engineering. Full article

The synthesis of various heterocyclic base modifications of nucleic acids has been thoroughly investigated; however, much less is known about their catabolism. Also, little is known about the transport of such compounds across the microbial cell membranes. Using the Saccharomyces cerevisiae single-gene deletion library, we performed genome-wide screening for genes affecting the growth of yeast in minimal media supplemented with N⁴-acetylcytosine as a source of uracil. We found that Fcy1, Fcy21, Bud16, Gnd1, and Fur4 proteins are required for efficient growth in the tested medium. Additionally, we used several heterocyclic pyrimidine bases and Fcy homolog mutants to test their growth in respective minimal media. We found that tested permeases differently affect the growth of yeast that is dependent on the heterocyclic pyrimidine bases used as a source of uracil. The most pronounced effect was observed for the ∆fur4 mutant, which was growing much slower than the corresponding wild-type strain in the media supplemented with N⁴-acetylcytosine, 4-methylthiouracil, N⁴-methylcytosine, N⁴,N⁴-dimethylcytosine, 2-thiouracil, or 4-thiouracil. We suggest that Fur4 protein is the major yeast transporter of modified heterocyclic pyrimidine bases. Our observations might be helpful when investigating the actions of various heterocyclic base-based antifungal, anticancer, and antiviral drugs. Full article

Wood is an important raw material for industrial applications. Its fiber-specific genetic modification provides an effective strategy to alter wood characteristics in tree breeding. Here, we performed a cross-analysis of previously reported single-cell RNA sequencing and the AspWood database during wood formation to identify potential xylem fiber-dominant expressing genes in poplar. As a result, 32 candidate genes were obtained, and subsequently, we further examined the expression of these genes in fibers and/or vessels of stem secondary xylem using the laser capture microdissection technique and RT-qPCR. Analysis identified nine candidate genes, including PtrFLA12-2, PtrIRX12, PtrFLA12-6, PtrMYB52, PtrMYB103, PtrMAP70, PtrLRR-1, PtrKIFC2-3, and PtrNAC12. Next, we cloned the promoter regions of the nine candidate genes and created promoter::GUS transgenic poplars. Histochemical GUS staining was used to investigate the tissue expression activities of these gene promoters in transgenic poplars. In one month, transgenic plantlets grown in medium showed intensive GUS staining signals that were visible in the leaves and apical buds, suggesting substantial expression activities of these gene promoters in plantlets predominantly undergoing primary growth. In contrast, for three-month-old transgenic poplars in the greenhouse with predominantly developed secondary stem tissues, the promoters of seven of nine candidate genes, including PtrMYB103, PtrIRX12, and PtrMAP70, showed secondary xylem fiber-dominant GUS signals with considerable spatial specificity. Overall, this study presents xylem fiber-dominant promoters that are well-suited for specifically expressing genes of interest in wood fibers for forest tree breeding. Full article

Turmeric (Curcuma longa L. cv. Trang 1), a high-value cultivar known for its elevated curcuminoid and volatile oil content, holds significant potential in pharmaceutical and food applications. However, its commercial propagation is constrained by low rhizome productivity and the limitations of conventional vegetative propagation. This study aimed to improve the propagation efficiency of turmeric cv. Trang 1 by developing optimized protocols for explant sterilization, shoot proliferation, root induction, and acclimatization. Sprouted rhizome buds were sterilized and cultured on a Murashige and Skoog (MS) medium supplemented with various plant growth regulators, including cytokinins (benzyladenine [BA], thidiazuron [TDZ], and meta-topolin [mT]) and auxins (indole-3-butyric acid [IBA] and 1-naphthaleneacetic acid [NAA]). The shoot induction (4.60 ± 1.47 shoots per explant) and shoot height (2.34 ± 0.61 cm) were observed on the MS medium with 3.0 mg/L BA, while the TDZ, at 0.5 mg/L, also induced a high number of shoots (5.22 ± 0.64). When using single shoots derived from bud explants, mT at 1.5 mg/L significantly enhanced the shoot formation. For the root induction, 2.0 mg/L IBA yielded the highest number of roots (7.33 ± 1.49), while NAA was less effective. The plantlets acclimatized in a 1:1 soil and peat moss mixture showed the highest survival rate (86.67%). This improved protocol enables the efficient production of turmeric plantlets, supporting commercial deployment. Full article

The leaf plastochron serves as an indicator of the rate of leaf appearance, biomass accumulation, and branch number, while also impacting plant architecture and seed yield. However, research on the leaf plastochron of crops remains limited. In this study, 2116C exhibited a rapid leaf plastochron compared to ZH18 during both rosette and bud periods. There were significant positive correlations among the leaf plastochron and primary branch number of the F₂ populations (r ranging from 0.395 to 0.635, p < 0.01). Genetic analyses over two years demonstrated that two equally dominant genes might govern the leaf plastochron. Through bulk segregant analysis sequencing (BSA-seq), three novel genomic intervals were identified on chromosomes A02 (9.04–9.48 Mb and 13.52–13.66 Mb) and A04 (19.84–20.14 Mb) of ZS11 and Darmor-bzh reference genomes. By gene functional annotations, single-nucleotide variation (SNV) analyses, transcriptome data from parents, genetic progeny, and natural accessions, we identified ten candidate genes within the intervals, including FLOWERING LOCUS T, RGL1, MYB-like, CYP96A8, BLH3, NIT2, ASK6, and three CLAVATA3/ESR (CLE)-related genes. These findings lay the molecular foundation for further exploration into the leaf plastochron and the implications in plastochron-related breeding in rapeseed. Full article

Chronic lower limb ischemia is a debilitating condition, particularly prevalent among elderly patients and individuals ineligible for revascularization procedures. Gene therapy aimed at promoting therapeutic angiogenesis presents a promising alternative treatment strategy. Objectives: This study evaluated the preclinical safety of a gene therapy drug composed of the plasmids pBudK-coVEGF-coANG and pBudK-coGDNF in laboratory animals. Safety assessment followed a single intramuscular injection at a dose 30 times higher than the proposed therapeutic level. Methods: Acute toxicity was monitored over a 24-h period. Genotoxicity was assessed using the micronucleus test at doses of 200, 1000, and 5000 μg/kg. Bone marrow cytology was analyzed to detect hematopoietic toxicity. Delayed toxicity was evaluated over a two-week recovery period. Results: No signs of acute toxicity were observed, even at the highest dose. The micronucleus test revealed no genotoxic effects, with no significant increase in micronucleated polychromatic erythrocytes compared to control groups. Bone marrow erythroblast parameters remained within normal physiological ranges. Additionally, no delayed adverse effects were detected during the recovery period. Conclusions: The gene therapy drug demonstrated a favorable preclinical safety profile, exhibiting no evidence of toxicity or genotoxicity, even at substantially elevated doses. These findings support the continued development of this therapy as a potential treatment for chronic lower limb ischemia in patients who are not candidates for surgical intervention. Full article

In order to explore the optimal vibration parameters for the selective harvesting of coffee fruits, a high-velocity dynamic photography monitoring system was developed to analyze the vibration-assisted harvesting process. This system identified the optimal vibration position on coffee branches and facilitated theoretical energy transfer analysis, obtaining a mathematical formula for calculating the total kinetic energy of coffee branches. A single-factor experiment was conducted with the vibration position as the experimental factor and the total kinetic energy of coffee branches as the response variable. The results showed that the total kinetic energy of the branches was the highest at Vibration Position 2 (the position between the third and the fourth Y-shaped bud tips on the branch). Therefore, Vibration Position 2 was determined as the optimal vibration position. Further analysis established a mathematical model linking coffee cherry motion parameters to theoretical detachment force. A factorial experiment was conducted with vibration frequency and amplitude as test factors, using detachment rates of green, semi-ripe, and ripe cherries as indicators. The results showed that at 55 Hz and 10.10 mm amplitude, the detachment rate of ripe cherries was highest (83.33%), while green and semi-ripe cherries detached at 16.67% and 33.33%, respectively. A field validation experiment, with Vibration Position 2, 55 Hz frequency, 10.10 mm amplitude, and 1 s vibration duration, yielded actual detachment rates of 15.86%, 35.17%, and 89.50% for green, semi-ripe, and ripe cherries, respectively. The error margins compared with the theoretical values were all below 10%. These results confirm the feasibility of optimizing vibration harvesting parameters through high-velocity photography dynamic analysis. Full article

The HIV-1 capsid is one of virology’s most iconic structures, yet how it assembles has long remained elusive. Remarkably, the capsid is made from just a single protein, CA, which forms a lattice of ~250 hexamers and exactly 12 pentamers. Conical capsids form inside budded virions during maturation, but early efforts to reproduce this in vitro resulted instead in open-ended tubes with a purely hexameric lattice. The missing component in capsid assembly was finally identified as the metabolite inositol hexakisphosphate (IP6). Simply mixing soluble CA protein with IP6 is sufficient to drive the spontaneous assembly of conical capsids with a similar size and shape to those inside of infectious virions. Equally important, IP6 stabilises capsids once formed, increasing their stability from minutes to hours. Indeed, such is the dependence of HIV-1 on IP6 that the virus actively packages it into virions during production. These discoveries have stimulated work from multiple labs into the role and importance of IP6 in HIV-1 replication, and is the subject of this review. Full article