Search Results (380)

Pancreatic ductal adenocarcinoma (PDA) exhibits diverse molecular aberrancies that contribute to its aggressive behaviour and poor patient survival. P-21-activated kinase 1 (PAK1) and PAK4 drive the tumorigenesis of PDA. However, their roles in tumour vasculature and the impact on immune response are unclear. This study aims to investigate the effects of PAK1 and PAK4 on tumour vasculature, immune cell infiltration, and the connection between using PAK1-knockout (KO), PAK4 KO, and wild-type (WT) PDA cells in cell-based and mouse experiments. Tumour tissues isolated from a syngeneic mouse model were immuno-stained to determine the changes in tumour vasculature and immune cell infiltration/activation, followed by a proteomic study to assess biological processes involved. PAK1KO or PAK4KO suppressed tumour growth by reducing angiogenesis while enhancing vascular normalisation, enhanced the infiltration/activation of T-cells and dendritic cells associated with upregulation of ICAM-1 and VCAM-1 in the tumour microenvironment, and stimulated vascular immune crosstalk via an ICAM-1-mediated mechanism. This was supported by proteomic profiles indicating the regulation of endothelial cell and leukocyte trans-endothelial migration in PAK1- or PAK4-knockout tumours. In conclusion, PAK1KO or PAK4KO enhanced tumour vascular normalisation while reducing angiogenesis, stimulating immune cell infiltration and activation to suppress tumour growth. Full article

Background: Osteosarcoma (OS) is a fast-growing malignant bone tumor that occurs most often in children and teenagers. Development of pulmonary metastasis is the primary cause of treatment failure and mortality. Our previous studies demonstrated that cytoplasmic p27 interacts with PAK1, enhancing PAK1 phosphorylation and promoting OS pulmonary metastasis. However, the cellular functions of p27 and PAK1 are primarily regulated by phosphorylation, and the roles of specific phosphorylation residues in modulating OS metastatic potential remain unclear. Methods: To study tumor invasiveness and lung metastasis, we employed a CRISPR-based knock-in method to introduce specific mutations—p27-T157A, p27-T157D, PAK1-T423E, and PAK1-K299R—into the 143B OS cell line, followed by in vitro invasion and orthotopic xenograft mouse experiments. These residues were selected for their therapeutic potential, as T157 regulates p27 nuclear–cytoplasmic shuttling, while T423 and K299 modulate PAK1 kinase activity. Results: No significant differences in pulmonary metastasis were observed across p27 mutants compared to parental controls. However, the p27-T157D mutant exhibited increased cytoplasmic mislocalization, elevated PAK1-S144 phosphorylation, and enhanced in vitro invasiveness compared to the p27-T157A mutant and parental 143B cells. The PAK1-K299R mutant, designed to be kinase-dead, showed negligible S144 phosphorylation, consistent with loss of kinase activity. Unexpectedly, this mutant displayed increased T423 phosphorylation and in vitro invasiveness, and significantly enhanced pulmonary metastasis in vivo compared to the PAK1-T423E mutant and parental controls. Conclusions: These findings highlight the complexity of targeting specific p27 and PAK1 phosphorylation sites as an anti-metastatic strategy for OS. While p27-T157 phosphorylation influences cytoplasmic localization and invasiveness, it does not significantly alter metastatic outcomes. Conversely, PAK1-T423 phosphorylation is critical in driving OS metastatic potential, and the kinase-dead K299R mutant’s unexpected pro-metastatic effect suggests that kinase-independent mechanisms or compensatory pathways may contribute to metastasis. Our findings suggest the necessity for a more comprehensive understanding of the phosphorylation dynamics of p27 and PAK1 in metastatic OS. They also indicate that conventional kinase inhibition may be insufficient and underscore the potential benefits of alternative or combinatorial therapeutic strategies, such as targeting kinase-independent functions or other upstream kinases involved in these regulatory pathways. Full article

Background: [⁶⁸Ga]Ga-DOTATOC is widely used in PET imaging of neuroendocrine tumors (NETs) due to its high affinity for somatostatin receptors. Given the short physical half-life of gallium-68 (~68 min), rapid, reproducible, and GMP-compliant synthesis is essential for clinical application. Methods: An optimized cassette-based automated synthesis protocol was developed using a commercial cassette. Improvements included direct generator elution into the reactor without pre-purification, use of a SepPak^® C18 Plus Light cartridge for purification, replacement of HEPES with 0.3 M sodium acetate buffer (final pH ~3.8), and implementation of a non-vented sterile filter enabling automated pressure-hold integrity testing. Results: Across all batches, the synthesis yielded [⁶⁸Ga]Ga-DOTATOC with high radiochemical purity (> 97%) and reproducible decay-corrected radiochemical yields up to 88.3 ± 0.6%. Total synthesis time was approximately 13 min. The final product remained stable for at least 3 h post-synthesis. The use of acetate buffer eliminated the need for HEPES-specific testing, streamlining the workflow. Automated filter testing improved GMP-compliant documentation and reduced radiation exposure for personnel. Conclusions: This optimized, cassette-based synthesis protocol enables fast, high-yield, and GMP-compliant production of [⁶⁸Ga]Ga-DOTATOC. It supports clinical theranostic workflows by ensuring product quality, process standardization, and regulatory compliance. Full article

The number of thoracolumbar vertebrae (NTLV) and the number of ribs (NR) are economically important traits in pigs due to their influence on carcass length and meat yield. Although VRTN is an established key gene, it fails to fully account for population-level variation in vertebral count, necessitating a further exploration of its genetic mechanisms. Given the efficacy of crossbred populations in mapping the genetic determinants of phenotypic variation, we analyzed 439 pigs from a Landrace × Yorkshire cross. Genotyping was performed via a 50 K SNP chip. Both NTLV and NR showed high heritability (0.700 and 0.752, respectively), while the number of lumbar vertebrae (NLV) showed limited variation (92.5% of pigs had NLV = 6). Using the BLINK model, four significant loci were identified. The most significant SNP, rs3469762345, located in the intergenic region between ABCD4 and VRTN, corresponds to a previously known QTL. Additionally, three novel variant sites (rs81211244, rs81347323, and rs81416674) were identified within or near the ALDH7A1, PTPRT, and PAK1 genes, which are known to play a role in bone development. This study uncovers novel swine candidate genes associated with vertebral and rib number variation, subsequently facilitating targeted research into their molecular mechanisms. Full article

The retrofit of semi-open transitional spaces in university buildings is essential for enhancing both thermal comfort and energy efficiency. However, most studies have focused on conventional indoor environments, overlooking the unique thermal characteristics of semi-open spaces and their impact on occupant comfort. This study integrated field measurements, occupant surveys, and AirPak simulations to develop a three-tier evaluation framework covering environmental parameters, subjective thermal perception, and simulation-based validation. Focusing on teaching buildings at Zhejiang University’s Zijingang Campus, the analysis revealed that the retrofit increased the daily mean air temperature by 2.1 °C and decreased the relative humidity by 3.6% in winter. The peak thermal comfort indices PET and PMV improved by 4.4 °C and 0.98, respectively, with a neutral PET identified at 13.3 °C. PMV showed a stronger correlation with TSV (p = 0.94, R² = 0.81) than PET. Simulations further validated the retrofit’s effectiveness in stabilizing the indoor thermal environment and reducing airflow discomfort. These findings provide both theoretical insights and practical guidance for the climate-responsive, energy-efficient retrofitting of campus buildings in hot summer and cold winter (HSCW) zones. Full article

Prostate cancer is a clinically heterogeneous disease. Since PSA is not cancer-specific, and due to the bone metastases seen in the advanced stage and bone deformations caused by hormone therapy, it is necessary to use new biomarkers. Formin-like-protein 1 (FMNL1), a member of the formin protein family, is of great importance in actin polymerization, cell attachment, and migration processes. p21-activated kinase 1 (PAK1) proteins, members of the PAK protein kinases, play a role in cytoskeletal organization, as well as regulating other cellular activities such as cell survival, mitosis, and transcription. In our study, plasma and urine samples of 60 prostate cancer patients and 20 healthy controls were studied using RT-PCR and ELISA methods. No statistical difference was found between FMNL1 mRNA and protein expression levels of patients and controls in both plasma and urine samples (p > 0.05). There was no statistical difference between PAK1 mRNA expression levels of patients and controls in plasma and urine samples (p > 0.05). While no significant difference was found in PAK1 protein levels in plasma samples (p > 0.05), it was found to be lower in urine samples of patients compared to the control group (p = 0.00). Both marker molecules have low expression levels in early-stage PCa. Full article

Developing environmentally friendly and cost-effective substrates is critical to enhance resource efficiency and productivity in plant factories with artificial lighting (PFALs). This study employed a molded coconut coir substrate (coconut coir composited with polyurethane hydrophilic adhesive, MCCS) in PFALs to cultivate lettuce (Lactuca sativa L.) and pak choi (Brassica rapa ssp. chinensis). During the transplanting stage, the roots exposed outside the MCCS of lettuce and pak choi were 13.40% and 19.92% shorter, respectively, than in the sponge treatment, and more amenable to mechanical transplanting. This compensated for the neglect of operational efficiency in traditional lifecycle assessment (LCA). Furthermore, compared with sponge and rockwool, MCCS significantly enhanced the yield of lettuce and pak choi by up to 27.33% and 67.19%, respectively. Meanwhile, MCCS significantly increased the chlorophyll content of lettuce compared to sponge by 8.56%. Compared with rockwool, MCCS significantly increased the chlorophyll b content (7.36%), antioxidant content, and antioxidant activity (total phenolics by 13.59%, total flavonoid by 18.43%, FRAP by 12.96%, and DPPH by 19.87%) of lettuce. For pak choi, MCCS increased the soluble protein content in the blade and total phenolics content in the petiole by 32.01% and 14.76%, respectively. More importantly, the use of MCCS led to a significant reduction in the energy consumption per unit area yield of lettuce and pak choi, with maximum reductions of 22.98% and 40.91%, respectively. This eco-friendly substrate is suitable for replacing sponge and rockwool in the production of lettuce and pak choi in PFALs. Full article

This study investigates growth responses, heavy metal (Cd, As) uptake, translocation, and mineral nutrient regulation in leafy vegetables with varying heavy metal tolerance, addressing the threat posed by combined Cd and As pollution. Three high-tolerance, four moderate-tolerance, and one sensitive leafy vegetable were grown in Cd+As-contaminated hydroponics. Post-harvest yields and concentrations of Cd, As, and trace elements were assessed. Results showed that (1) compared with single heavy metal treatments, the combination of Cd and As significantly increased the translocation factor of Cd in black bean sprouts and white radish sprouts by up to 83.83% and 503.2%; (2) changes in mineral nutrient concentrations in leafy vegetables were similar between single and combined heavy metal stresses, but the regulatory patterns varied among different leafy vegetable species; (3) under Cd/As exposure, high-tolerance leafy vegetables (e.g., pak choi) had strong heavy metal accumulation abilities, and heavy metal stress positively regulated mineral elements in their roots; In contrast, sensitive leafy vegetables (e.g., pea sprouts) often exhibited suppressed mineral element content in their roots, which was a result of their strategy to reduce heavy metal uptake. These results offer key insights into resistance mechanisms against combined heavy metal pollution in leafy vegetables, supporting phytoremediation efforts and safe production. Full article

In recent years, the region of Valparaíso has faced devastating fires, notably the Viña del Mar fire on 2 February 2024, which affected 9252 hectares. This study analyzes fire behavior in informal settlements and assesses the effectiveness of different construction materials through scaled prototypes of dwellings made from MDF, OSB, TetraPak, and flame-retardant resin composites. Controlled fire experiments were conducted, recording fire spread times and atmospheric conditions. Results confirm significant differences in fire spread rates and structural survival times between materials, highlighting the practical benefit of fire-resistant alternatives. The Kaplan–Meier survival analysis indicates critical time thresholds for rapid flame escalation and structural collapse under semi-open conditions, supporting the need for improved safety measures. Burn pattern observations further revealed the role of wind, thermal radiation, and material properties in fire dynamics. Overall, this study provides experimental evidence aligned with real fire scenarios, offering quantified insights to enhance fire prevention and response strategies in vulnerable settlements. These findings provide an exploratory basis for understanding fire dynamics in informal settlements but do not constitute definitive design prescriptions. Full article

Microgreens, having short growth cycles and efficient nutrient uptake, are ideal candidates for biofortification. This study investigated the effects of selenium (Se) and zinc (Zn) on photosynthetic performance in four hydroponically grown Brassica microgreens (broccoli, pak choi, kohlrabi, and kale), using direct and modulated chlorophyll a fluorescence and chlorophyll-to-carotenoid ratios (Chl/Car). The plants were treated with Na₂SeO₄ at 0 (control), 2, 5, and 10 mg/L or ZnSO₄ × 7H₂O at 0 (control), 5, 10, and 20 mg/L. The results showed species-specific responses with Se or Zn uptake. Selenium enhanced photosynthetic efficiency in a dose-dependent manner for most species (8–26% on average compared to controls). It increased the plant performance index (PI_tot), particularly in pak choi (+62%), by improving both primary photochemistry and inter-photosystem energy transfer. Kale and kohlrabi exhibited high PSII-PSI connectivity for efficient energy distribution, with increased cyclic electron flow around PSI and reduced Chl/Car up to 8.5%, while broccoli was the least responsive. Zinc induced variable responses, reducing PI_tot at lower doses (19–23% average decline), with partial recovery at 20 mg/L (9% average reduction). Broccoli exhibited higher susceptibility, with inhibited Q_A re-oxidation, low electron turnover due to donor-side restrictions, and increased pigment ratio (+3.6%). Kohlrabi and pak choi tolerated moderate Zn levels by redirecting electron flow, but higher Zn levels impaired PSII and PSI function. Kale showed the highest tolerance, maintaining stable photochemical parameters and total electron flow, with increased pigment ratio (+4.5%) indicating better acclimation. These results highlight the beneficial stimulant role of Se and the dual essential/toxic nature of Zn, thus emphasizing genotype and dose-specific optimizations for effective biofortification. Full article

This study aimed to investigate the underlying mechanisms by which dandelion extract inhibits the proliferation of breast cancer MDA-MB-231 cells. Dandelion root and leaf extracts were prepared using a heat reflux method and subjected to solvent gradient extraction to obtain fractions with different polarities. MTT assays revealed that the ethyl acetate fraction exhibited the strongest inhibitory effect on cell proliferation. LC-MS analysis identified 12 potential active compounds, including sesquiterpenes such as Isoalantolactone and Artemisinin, which showed significantly lower toxicity toward normal mammary epithelial MCF-10A cells compared to tumor cells (p < 0.01). Mechanistic studies demonstrated that the extract induced apoptosis in a dose-dependent manner, with an apoptosis rate as high as 85.04%, and significantly arrested the cell cycle at the S and G2/M phases. Label-free quantitative proteomics identified 137 differentially expressed proteins (|FC| > 2, p < 0.05). GO enrichment analysis indicated that these proteins were mainly involved in cell cycle regulation and apoptosis. KEGG pathway analysis revealed that the antitumor effects were primarily mediated through the regulation of PI3K-Akt (hsa04151), JAK-STAT (hsa04630), and PPAR (hsa03320) signaling pathways. Moreover, differential proteins such as PI3K, AKT1S1, SIRT6, JAK1, SCD, STAT3, CASP8, STAT2, STAT6, and PAK1 showed strong correlation with the core components of the EA-2 fraction of dandelion. Molecular docking results demonstrated that these active compounds exhibited strong binding affinities with key target proteins such as PI3K and JAK1 (binding energy < −5.0 kcal/mol). This study elucidates the multi-target, multi-pathway synergistic mechanisms by which dandelion extract inhibits breast cancer, providing a theoretical basis for the development of novel antitumor agents. Full article

Chronic myeloid leukemia (CML) is a leading example of a malignancy where a molecular targeted therapy revolutionized treatment but has rarely led to cures. Overcoming tyrosine kinase inhibitor (TKI) drug resistance remains a challenge in the treatment of CML. We have recently identified miR-185 as a predictive biomarker where reduced expression in CD34⁺ treatment-naïve CML cells was associated with TKI resistance. We have also identified PAK6 as a target gene of miR-185 that was upregulated in CD34⁺ TKI-nonresponder cells. However, its role in regulating TKI resistance remains largely unknown. In this study, we specifically targeted PAK6 in imatinib (IM)-resistant cells and CD34⁺ stem/progenitor cells from IM-nonresponders using a lentiviral-mediated PAK6 knockdown strategy. Interestingly, the genetic and pharmacological suppression of PAK6 significantly reduced proliferation and increased apoptosis in TKI-resistant cells. Cell survivability was further diminished when IM was combined with PAK6 knockdown. Importantly, PAK6 inhibition in TKI-resistant cells induced cell cycle arrest in the G2-M phase and cellular senescence, accompanied by increased levels of DNA damage-associated senescence markers. Mechanically, we identified a PAK6-mediated MDM2-p21 axis that regulates cell cycle arrest and senescence. Thus, PAK6 plays a critical role in determining alternative cell fates in leukemic cells, and targeting PAK6 may offer a therapeutic strategy to selectively eradicate TKI-resistant cells. Full article

Salinization severely impairs crop growth by inducing oxidative stress and disrupting cellular homeostasis. This study systematically investigates the synergistic effects of salt-tolerant plant-growth-promoting rhizobacteria (ST-PGPR) and foliar silicon fertilizer spraying (FSFS) on antioxidant responses in Pak choi under salt stress. Two-season pot experiments were carried out to evaluate key indicators, including antioxidant enzyme activities (superoxide dismutase: SOD; peroxidase: POD; catalase: CAT), oxidative stress (malondialdehyde: MDA), osmolyte accumulation (proline, soluble protein), and hormones (Jasmonic Acid: JA; Salicylic Acid: SA; Abscisic acid: ABA). The results demonstrate that combining ST-PGPR with FSFS significantly enhances SOD (6.18–2353.85%), POD (3.44–153.29%), and CAT (25.71–319.29%) activities while reducing MDA content (8.12–35.87%). Proline and soluble protein levels increased by 1.56–15.71% and 5.03–188.87%, respectively. Hormonal regulation increased JA, SA, and ABA levels by 1.05–31.81%, 2.09–34.29%, and 3.18–30.09%, respectively. Notably, ST-PGPR treatments at 10⁴ and 10⁶ cfu·mL⁻¹, combined with foliar silicon application, consistently ranked highest in overall antioxidant performance across both seasons based on a principal component analysis. These findings provide novel insights into microbial–mineral interactions for sustainable saline agriculture. Full article

Background: While the mechanism of asymmetric gonadal development is generally understood, the mechanism of asymmetric oviduct development remains unclear. Methods: Right and left oviducts were collected from chick embryos at three developmental stages (Embryonic day 7.5, E9.5, and E11.5) for RNA-seq analysis (RNA-seq). Whole-genome resequencing (WGRS) was performed on hens with bilateral reproductive systems (a rare natural occurrence) and unilateral controls. These data were co-analyzed with public RNA-seq data of female embryonic gonads at different developmental stages (E4.5, E5.5, and E6.5) to screen for candidate genes affecting oviduct degeneration/development. Results: RNA-seq analyses showed that a total of 27, 10, and 38 DEGs were identified between the left and right oviducts at E7.5, E9.5, and E11.5, respectively. WGRS analyses revealed 1045 differentially mutated genes (DMGs) between bilateral (D) and unilateral (S) groups. Preliminary validation highlighted BMP7, PAK3, SLC6A11, PITX2, and SMC1B as candidate genes influencing oviduct asymmetry. Conclusions: This study provides insights into the genetic basis of asymmetric oviduct development and lays the groundwork for breeding hens with bilateral reproductive systems. Full article

Health and wellness tourism is a rapidly expanding segment of the global tourism industry, driven by increasing consumer awareness of well-being and lifestyle enhancement. As the demand for wellness travel grows, destinations are expected to offer high standards of safety, hygiene, rehabilitation, and holistic experiences. This study aims to identify and evaluate the key attributes and determinants for developing health and wellness tourism destinations by applying the 6As Tourism Development framework: Attractions, Accessibility, Amenities, Activities, Available Packages, and Ancillary Services. A multi-criteria decision-making approach, specifically the TOPSIS, was employed to assess destination potential through a case study of Nakhon Ratchasima Province, Thailand. The results indicate that Attractions, Accessibility, and Amenities are the top three priorities for wellness tourists. Sub-criteria such as natural scenery, cultural significance, accessibility for all, safety, and accommodation quality are particularly influential. Three districts in Nakhon Ratchasima were found to exhibit distinct strengths—Pak Chong is best suited for rehabilitative tourism (e.g., aroma and water therapy), aligning with mind and nutrition wellness components; Wang Nam Khiao is ideal for ecotourism and cultural experiences, supporting environmental and nutritional dimensions; while Mueang Nakhon Ratchasima excels in sports tourism, supporting physical and nutritional well-being. The study offers practical insights for policymakers and tourism stakeholders to design sustainable, visitor-centered wellness destinations. The proposed framework supports strategic planning and resource allocation for health-focused tourism development. Full article