Search Results (929)

The salivary gland is a key organ in insects that plays essential roles in food digestion, nutrient absorption, and energy metabolism, thereby highlighting the importance of studying salivary gland function for gaining a better understanding of nutritional utilization and insect–plant interactions. To date, however, a lack of salivary gland-specific promoters has limited functional analyses of salivary gland genes in Lepidoptera. In this study, based on microarray and salivary gland transcriptome data, we identified nine candidate genes characterized by high salivary gland expression. Semi-quantitative PCR analysis confirmed cholinesterase (BmCe, BGIBMGA010988) as the optimal candidate for promoter cloning. Temporal expression analysis revealed that the expression of BmCe reaches a peak during days 2–4 of the fifth larval instar. A 2152 bp fragment upstream of the transcription initiation site of BmCe was selected as the putative promoter sequence (designated BmCeP) and cloned to construct a piggyBac transgenic vector driving DsRed expression. Transgenic silkworms were obtained via embryonic microinjection and tissue expression analysis on day three of fifth-instar larvae revealed the predominant localization of DsRed expression in the salivary glands. In this study, we thus identified a gene promoter characterized by salivary gland-predominant expression in Bombyx mori, which we believe could serve as a valuable genetic tool for investigating the molecular mechanisms underlying silkworm nutritional utilization and interactions with its host plant, mulberry. Full article

Introduction: Hypervirulent Klebsiella pneumoniae (hvKp) is an emerging global pathogen that causes severe liver abscesses and metastatic infections. Despite rising concerns regarding multidrug-resistant convergence, molecular data in Romania remain limited. This study characterizes the epidemiological, clinical, and genomic profiles of hvKp liver abscesses in a tertiary hospital, aiming to describe the local virulence landscape and inform clinical management. Results: This study characterizes 15 cases of hvKp liver abscesses in a tertiary hospital. The cohort was predominantly male, with high rates of type 2 diabetes. Although clinical presentation was severe, featuring complications like endogenous endophthalmitis, the outcomes following prolonged antibiotic therapy were generally favorable. Phenotypically, 93.3% of isolates retained a wild-type susceptible profile, while a single ESBL-positive case highlighted the risk of resistance convergence. Genomic sequencing confirmed the presence of the ST23-K1 pandemic lineage carrying key virulence determinants (rmpA, iuc, and peg-344). Crucially, cgMLST analysis revealed genetic heterogeneity, suggesting sporadic community acquisition rather than a clonal nosocomial outbreak. Conclusions: These findings represent the first detailed molecular description of hvKp in Romania, confirming the local circulation of high-risk ST23 clones and underscoring the necessity for early detection and surveillance. Full article

Prolyl 3-hydroxylase 2 (P3H2) is a key enzyme involved in the architecture of the extracellular matrix (ECM). While previously shown to be regulated by VEGF-A and to play a role in angiogenesis, its function in cancer remains ambiguous. While characterized as a tumor suppressor, its precise function in colorectal cancer (CRC) progression is poorly defined. Bioinformatic analysis and patient data reveal that P3H2 transcript levels are significantly reduced in colon adenocarcinoma tissues, showing a progressive decline in metastatic lesions. Furthermore, VEGF-A exposure upregulates P3H2 transcripts in the HCT116 CRC cell line. To investigate its impact in CRC, we generated a stable HCT116 clone overexpressing P3H2. In vitro studies demonstrated that while P3H2 overexpression inhibited anchorage-independent growth, it significantly enhanced cellular invasion without altering cell proliferation. In vivo, however, P3H2-overexpressing tumors exhibited accelerated tumor growth and a statistically significant increase in lung metastases. P3H2 overexpression remodeled the tumor microenvironment (TME) by modifying its main substrate, Collagen IV, resulting in the induction of increased vessels density. Our study repositions P3H2 as a dynamic enzymatic switch within the TME. This work identifies P3H2-driven ECM remodeling as a promising therapeutic axis in advanced CRC, with particular relevance for combination strategies targeting angiogenesis. Full article

PIM kinases (PIM1, PIM2, PIM3) are serine/threonine kinases implicated in infection and reactivation of various viruses, but their roles in HIV-1 gene expression and particle production remain unclear. We examined their impact on HIV-1 and related viruses using co-transfection systems. PIM1 and PIM3, but not PIM2, markedly suppressed HIV-1 virion production without affecting infectivity. This inhibitory effect extended to transmitted/founder HIV-1 clones and SIV, indicating broad activity across lentiviruses. Kinase-dead mutants failed to reduce virion production, confirming the requirement for catalytic activity. Our data suggest that PIM1 and PIM3 act at distinct steps of HIV-1 gene expression: PIM1 reduces transcription, whereas PIM3 acts post-transcriptionally to diminish viral protein expression. Co-expression of PIM1 and PIM3 further enhanced suppression, suggesting complementary functions. Both kinases also inhibited expression from non-LTR promoters, implying involvement of general cellular factors. These findings reveal distinct and cooperative actions of PIM1 and PIM3 in limiting HIV-1 particle production, providing new insights into host kinase-mediated regulation of viral gene expression. Full article

Black locust (Robinia pseudoacacia L.) breeding is an important component of plantation forestry in Central and Eastern Europe; however, clone trials are still mainly evaluated using conventional field surveys, and the application of high-resolution red-edge satellite indices at the intraspecific level remains rarely applied. As a result, less information is available on the phenological status of black locust clones derived from red-edge satellite data. This study evaluates a clone trial established in Eastern Hungary on slightly acidic Arenosol soil, assessing the growth performance and seasonal spectral dynamics of newly bred black locust clones during their fifth growing season by integrating field measurements with PlanetScope-derived Normalized Difference Red-Edge Index (NDRE) time series. Clone NK2 exhibited the most vigorous growth, reaching a mean height of 11.1 ± 0.15 m and a diameter at breast height (DBH) of 11.21 ± 0.19 cm, which were 35.4% greater in height and 19.0% larger in DBH than those (8.2 ± 0.12 m height, and 9.42 ± 0.23 cm diameter) of the control (‘Üllői’ cultivar). Clone PL251 also exceeded the control by 25.6% in height and 19.2% in DBH. Spectral analysis (NDRE value ± standard error) revealed marked differences in phenological development: in the early stage (April 15), NK1 and PL040 had the highest NDRE values (0.472 ± 0.020 and 0.461 ± 0.019), whereas NK2 showed delayed leaf emergence (0.398 ± 0.019). By June 21, PL251 had reached an NDRE value of 0.692 ± 0.013, which was higher than that of the control (0.673 ± 0.016). In mid-July, NDRE peaked for NK2 and NK1 (0.732 ± 0.012 and 0.731 ± 0.013), with ‘Üllői’ showing consistently lower values across the season. In the final stage, NK2 maintained the highest NDRE values (October 22: 0.618 ± 0.015; November 9: 0.466 ± 0.021), indicating prolonged photosynthetic activity, while NK1 and ‘Üllői’ declined earlier (e.g., November 9: 0.354 ± 0.018 and 0.390 ± 0.027, respectively). These findings highlight NK2 and PL251 as superior candidates for high-yield, climate-resilient tree plantations because of their strong growth and extended physiological activity. Full article

The rise of SIM cloning, identity spoofing, and covert manipulation in mobile and IoT networks has created an urgent need for continuous post-registration verification. This work introduces an unsupervised deep learning framework for detecting behavioral anomalies in SIM-tagged network flows by modeling the intrinsic structure of benign behavioral descriptors (TTL, timing drift, payload statistics). A Temporal Deep Autoencoder (TDAE) combining Conv1D layers and an LSTM encoder is trained exclusively on normal traffic and used to identify deviations through reconstruction error, enabling one-class (label-free) training. For deployment, alarms are set using an unsupervised quantile threshold ${\tau }_{\alpha }$ calibrated on benign traffic with a false-alarm budget; ${\tau }^{*}$ is reported only as a diagnostic reference for model comparison. To ensure realism, a large-scale corpus of 3.6 million SIM-tagged flows was constructed by enriching public IoT traffic with pseudo-operator identifiers (synthetic SIM tags derived from device identifiers) and controlled anomaly injections. Cross-domain experiment transfer under SIM-grouped protocol: Training on clean Cassavia-like traffic and testing on attack-rich Guarascio-like flows yields a PR-AUC of 0.93 for the proposed Conv-LSTM Temporal Deep Autoencoder, outperforming Dense Autoencoder, Isolation Forest, One-Class SVM, and LOF baselines. Conversely, the reverse direction collapses to PR-AUC $\approx 0.5$, confirming the absence of data leakage and the validity of one-class behavioral learning. Sensitivity analysis shows that performance is stable around the unsupervised quantile operating point. Overall, the proposed framework provides a lightweight, interpretable, and data-efficient behavioral verification layer for detecting cloned or unauthorized SIM activity, complementing existing registration mechanisms in next-generation telecom and IoT ecosystems. Full article

Richter transformation (RT) represents a rare but highly lethal evolution of chronic lymphocytic leukemia/small lymphocytic lymphoma (CLL/SLL), most frequently manifesting as diffuse large B-cell lymphoma (DLBCL). Despite therapeutic advances in CLL, DLBCL-RT remains characterized by rapid progression, profound treatment refractoriness, and short survival with conventional chemoimmunotherapy, underscoring the need for a refined biological and therapeutic framework. A defining feature of RT is clonal relatedness: most cases arise through linear or branched evolution of the antecedent CLL clone and carry an inferior prognosis compared with clonally unrelated cases that resemble de novo DLBCL. Recent multi-omic data further indicate that clonally related RT commonly originates from minute, transformation-primed subclones detectable years before clinical emergence, shifting RT from a late stochastic event to an early-established evolutionary trajectory. At transformation, recurrent genetic lesions of TP53, CDKN2A/B, NOTCH1, and MYC cooperate with B-cell receptor-associated programs, epigenetic reconfiguration, and metabolic rewiring toward OXPHOS- and mTOR-driven states, collectively promoting genomic instability and aggressive growth. In parallel, RT develops within a profoundly immunosuppressive microenvironment marked by PD-1-expressing malignant B cells, PD-L1-rich myeloid niches, exhausted T cells, expanded regulatory T cells, and M2-skewed macrophages interconnected by redundant checkpoint and cytokine networks. Therapeutic strategies are rapidly evolving, including pathway inhibitors, immune checkpoint blockade, T-cell-engaging bispecific antibodies, CAR-T therapies, and antibody–drug conjugates. This review integrates current insights into RT pathogenesis, immune escape, and emerging therapies, highlighting opportunities for biomarker-driven patient stratification, rational combinations, and earlier interception of transformation-prone disease. Full article

Molecular genotyping is a key factor for plant breeding programming and plant variety protection (PVP). However, its potential still remains to be elucidated when considering ornamental plants like Petunia × hybrida. In this study, a petunia breeding clone collection, including sister line groups, was genotyped through double digest Restriction-site Associated DNA sequencing (ddRADseq), and its genetic diversity and structure were studied. In addition to estimating the high genetic similarity observed among sister lines, this approach allowed the unique discrimination of each clone too. Molecular results agreed with genealogy data, supporting the assessment of genotyping effectiveness. In addition, the minimal number of variants able to uniquely discriminate and/or correctly cluster the experimental lines was investigated. The loci number could be reduced to eight to achieve line discrimination, and a method to identify the specific variant sets is presented. Conversely, to preserve the original clustering with minor adjustments, one hundred loci were required and were obtained through minor allele frequency (MAF) filtering. Moreover, analysis of the chromosomal distribution of variants revealed a predominant accumulation in distal regions. Genetic analyses were repeated considering only variants located in coding sequences and results were in agreement with what previously observed, disclosing the potential of the expressed regions for genotyping purposes. Eventually, the applied approach enabled the investigation of SNPs within genes putatively involved in traits of interest. Our findings encourage the adoption of high-throughput and cost-effective sequencing techniques for petunia genotyping aimed at achieving PVP, supporting new variety registration, and developing marker-assisted breeding (MAB) and marker-assisted selection (MAS) strategies. Full article

Background/Objectives: Low-grade serous ovarian carcinoma (LGSOC) is known as an uncommon subtype of cancer with poor response to standard chemotherapy, so novel targets are required. The current study aims to highlight the Human Epidermal Growth Factor Receptor-2 (HER2/neu) immunohistochemical (IHC) expression in LGSOCs. Methods: The study was conducted using 33 cases of LGSOCs from the calendar years 2017–2024. IHC staining was performed using antibody anti-HER2/neu (clone 4B5). HER2/neu scoring was performed based on the American Society of Clinical Oncology and the College of American Pathologists (ASCO/CAP) criteria for breast carcinoma. Results: The mean age of the 33 patients was 46.5 years. International Federation of Gynecology and Obstetrics (FIGO) stage data for patients revealed a predominance of advanced disease: 82.7% (24/29) were in advanced stages. Early stages comprised 17.3% (5/29) of cases. The study did not find HER2/neu overexpression in all cases. In 3.0% of samples (1/33), HER2/neu IHC staining was scored as 1+, and in 6.1% (2/33) of all LGSOCs, ultralow phenotype was observed. Of 23 cases in the HER2-negative group, 6 patients were alive with progressive disease, 1 patient died in 5 months, and 16 were alive with no evidence of disease. Of two patients with the HER2-ultralow phenotype, one was alive with no evidence of disease at 16 months follow-up. Conclusions: The results support the idea that HER2/neu overexpression is exceptionally rare in LGSOC; nevertheless, future trials are essential to fully characterize the spectrum of HER2/neu alterations in LGSOC and to determine definitively whether the rare cases with mutations or ultralow expression could represent a small subgroup that might benefit from specific targeted agents. Full article

Background/Objectives: Pseudomonas aeruginosa is an opportunistic pathogen frequently implicated in healthcare-associated infections, particularly ventilator-associated pneumonia and other device-related infections. The global emergence of carbapenem-resistant P. aeruginosa (CRPA) represents a major clinical challenge due to its limited therapeutic options and high mortality rates. Methods: Relevant clinical data were obtained from medical records. Isolates were identified via 16S PCR, and antimicrobial susceptibility testing was performed using the Vitek2 Compact system following CLSI guidelines. Carbapenemase genes (blaGES, blaKPC, blaIMP, blaNDM, blaVIM) were detected via PCR. Clonal relationships were determined via RAPD-PCR, and some sequence types were assigned according to the global P. aeruginosa MLST database. Results: In this study, 40 non-duplicate CRPA isolates were collected from 35 patients in a tertiary-care hospital in Mexico. Most isolates originated from adult patients, predominantly from tracheal aspirates (32.5%) and urine cultures (25.0%). Mechanical ventilation was the most common invasive device associated with infection, and the overall mortality rate reached 14.3%. Antimicrobial susceptibility testing showed that 95% of isolates exhibited a multidrug-resistant phenotype, with high resistance rates to ciprofloxacin (70.0%) and β-lactams. Carbapenemase genes were detected in 55% of isolates, mainly bla_IMP, bla_GES, and bla_VIM, either alone or in combination. Notably, this is the first report of ST309 (bla_IMP), ST411 (bla_GES + bla_IMP), and ST167 (bla_GES + bla_VIM) carrying carbapenemase genes in Mexico. Conclusions: These findings highlight the persistence and genetic diversity of CRPA circulating in hospital settings and emphasize the urgent need for strengthened genomic surveillance and infection control programs to prevent the spread of these high-risk multidrug-resistant clones. Full article

This research simulates Aswan City’s urban transportation dynamics utilizing the Multi-Agent Transport Simulation (MATSim) framework. As a fast-expanding urban center, Aswan has many transportation difficulties that require extensive modeling toward sustainable mobility solutions. MATSim, recognized for its agent-based methodology, offers a detailed portrayal and analysis of individual travel behaviors and their interactions within the metropolitan transportation system. This study compiled and combined many databases, including demographic data, road infrastructure, public transit plans, and travel demand trends. These data are altered to produce a realistic digital clone of Aswan’s transportation system. Simulated scenarios analyze the consequences of several actions, such as increased public transit scheduling, traffic flow management, and the adoption of alternative transport modes, on minimizing congestion and boosting accessibility. Pilot findings show that MATSim effectively captures the distinct features of Aswan’s transportation network and offers practical insights for decision-makers. The results identified some opportunities to improve mobility and promote sustainable urban growth in developing cities. This study emphasized the importance of agent-based simulations in designing future transportation systems and urban infrastructure. Full article

Streptococcus pneumoniae (S. pneumoniae) is responsible for a wide range of infections. The aim of this study was to investigate the clonal diversity of S. pneumoniae in thirteen Arab countries. Multi-Locus Sequence Typing (MLST) data were extracted from PubMLST database. Genetic analysis was performed using DnaSP software version 6.0. A Minimum Spanning Tree (MST) analysis was conducted to evaluate the population structure of S. pneumoniae strains. Genetic data from 1008 Arab S. pneumoniae strains, collected over 22 years (1996–2018), were analyzed. MLST analysis identified a highly diverse population comprising 600 sequence types grouped into 87 clonal complexes and 295 singletons. Both internationally disseminated clones (e.g., ST156) and country-specific lineages (e.g., ST2307, Saudi Arabia) were observed, indicating substantial geographic structuring. Significant associations were detected between sequence types and geographical origin, decade of isolation, patient age, disease type, and serotype (p < 0.05). Although recombination events were presented, the population retained a predominantly clonal structure over time (I^S_A = 0.0715, p < 0.001). Overall, these findings demonstrated extensive genetic heterogeneity and spatiotemporal structuring of S. pneumoniae in the Arab region, providing valuable insights for regional surveillance and vaccine-related strategies. Full article

Pig somatic cell nuclear transfer (SCNT) has valuable applications in agriculture, biomedicine, and life sciences, yet low cloning efficiency remains a major constraint limiting its application. To systematically investigate factors related to the production efficiency of pig cloning, this study conducted a retrospective analysis of 367,701 SCNT embryos transferred into 2019 surrogate sows over five years, focusing on breeds of donor cells, the season of embryo transfers, and the number of embryos transferred per surrogate. Our data demonstrate that the genetic background of donor cells is a critical determinant. SCNT embryos generated by wild-type (WT) Pietrain and Duroc pigs yielded significantly higher cloning efficiencies compared to those from Large White and Yorkshire pigs. This breed-specific influence was also observed with genetically modified (GM) donor cells. Nevertheless, within the GM groups, GM-Duroc and GM-Yorkshire showed superior efficiency compared to GM-Large White and GM-Bama. Furthermore, Summer was identified as the least favorable season for embryo transfer, with significantly lower pregnancy rates, delivery rates, and cloning efficiency compared to the other seasons. Importantly, we established that transferring 100–150 embryos per recipient optimized cloning efficiency, significantly outperforming groups receiving higher embryo numbers without compromising pregnancy rates, delivery rates, or average litter sizes. Our findings provide valuable guidance for optimizing large-scale SCNT protocols in swine. Full article

Path planning in complex, dynamic environments presents a significant challenge. Deep Reinforcement Learning (DRL) offers an end-to-end solution but suffers from critical sample inefficiency and a “cold-start” problem. Imitation Learning (IL) accelerates training but is constrained by a performance ceiling and poor generalization. To address these limitations, we propose a novel Imitation–Reinforcement Hybrid Machine Learning Algorithm (Hybrid IL-RL). This framework balances exploration and performance via a two-stage process: First, an offline pre-training phase uses Behavioral Cloning (BC) with “non-expert” A* data from static environments for a “warm start”. Second, an online fine-tuning phase uses a DRL algorithm (SAC) to adapt this policy in complex, dynamic environments, allowing the agent to surpass the teacher’s limitations. Simulation experiments validate the approach. The framework demonstrates significantly faster convergence than DRL algorithms trained from scratch. Most critically, in the dynamic environment, our Hybrid IL-RL algorithm achieved the highest success rate (82.4%), while pure IL methods (BC, GAIL) failed due to poor generalization (e.g., 82.1% collision rate) and pure DRL methods struggled (approx. 51–56% success rate). Our results confirm the hybrid framework effectively solves the cold-start problem while using DRL to break the IL performance ceiling. Full article

Rhododendrons naturally inhabit acidic soils where aluminum (Al) toxicity severely restricts plant growth, yet the molecular basis underlying cultivar-dependent differences in Al tolerance remains poorly understood. In this study, we compared the transcriptional and physiological responses of an Al-resistant cultivar (Kangnaixin) and an Al-sensitive cultivar (Baijinpao) under Al stress. Transcriptome analysis was performed to identify Al-responsive differentially expressed genes (DEGs), followed by Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses to elucidate functional categories and metabolic pathways involved in stress adaptation. In addition, the Al tolerance-related gene RsALS3 was cloned and functionally characterized through heterologous overexpression in Arabidopsis thaliana. The two cultivars exhibited distinct transcriptional profiles in response to Al stress, with DEGs significantly enriched in abiotic stress responses, membrane-associated functions, and key metabolic pathways, including starch and sucrose metabolism, phenylpropanoid and flavonoid biosynthesis, and photosynthesis-related processes. These results suggest that Al stress disrupts membrane integrity and alters carbon metabolism in Rhododendron. Functional validation demonstrated that RsALS3 overexpression moderately alleviated Al-induced toxicity in A. thaliana, as evidenced by reduced leaf damage and improved photosynthetic efficiency. Although the observed phenotypic differences were modest, and some chlorophyll fluorescence kinetics data did not reach strong statistical significance. The overall physiological trends support a potential role of RsALS3 in Al stress adaptation. Collectively, these findings provide insight into cultivar-specific Al stress responses in Rhododendron and identify RsALS3 as a promising candidate gene for further investigation aimed at improving adaptation to acidic soils. Full article