Search Results (93)

Both let-7a and miR-34a have been repeatedly studied as pivotal suppressors for Hepatocellular carcinoma; however, their combined regulations remain to be fully elucidated. In the present study, we performed a comprehensive in silico analysis for let-7a and miR-34a using a wealth of updated tools: miRWalk, Genetrail and miRnet. In addition, our study is the first to quantify both miRs and their three predicted yet not experimentally validated oncogenic targets: FNDC3B, IGF2 and SOX4. This was assessed in HepG2 cell model following treatment by PEGP-vector expressing the miRs by MTT assay, florescence microscopy, qPCR and immune-florescence. Our bioinformatics analysis revealed a pool of common predicted hepatocarcinogenic targets shared by both let-7a and miR-34a. Importantly, three targets were identified as co-regulated through multiple canonical binding sites for each miR, and these had not been experimentally validated before. Furthermore, functional enrichment of these putative targets demonstrated their significant involvement in major and emerging HCC hallmarks, such as reprogramming of energy metabolism and evading immune destruction. These findings support our concept of simultaneous co-regulation of these oncogenes through the signaling networks and GO terms associated with both miRs. Consistently, our experimental results verified the significant overexpression of both miRs in HepG2 cells, leading to reduced tumor cell proliferation and decreased levels of the three oncogenic transcripts. Interestingly, miR-34a exhibited a superior suppression effect, reaching 38.7%, and SOX4 was identified as the most significantly downregulated target at both transcriptional and translational levels. Our findings provide new insights into the interconnected anti-HCC effects of let-7a and miR-34a and highlight the potential of applying their combined use to achieve the best therapeutic outcomes for this invasive tumor. Full article

Background: The standard diagnostic approach for prostate cancer (PCa) diagnosis consists of serum prostate-specific antigen (PSA) testing, digital rectal examination (DRE) and image-guided targeted biopsies. Given the invasive nature, potential adverse events and costs associated with these techniques, alternative approaches have been investigated, specifically with serum and urine assays. The work presented here is intended to further validate a novel noninvasive optical technique for PCa detection, targeting the VPAC genomic receptors that are overexpressed on prostate cancer’s malignant cells (MC), in non-DRE voided urine. Methods: Patients (N = 62) who had image-guided biopsy and histologically confirmed localized PCa, and who were scheduled for radical prostatectomy, provided a non-DRE voided urine sample prior to surgery. Urine was cytocentrifuged and cells fixed on a glass slide, incubated with 0.5 μg TP4303 (a receptor-specific fluorophore developed in our laboratory with high affinity for VPAC), excess washed and treated with 4,6-diamidodino-2-phenylindole (DAPI) for nuclear staining. The field of cells on each slide was analyzed using a Zeiss AX10 Observer microscope (20×). The total number of cells and MC were then counted, and the florescent intensity around each MC was measured using Zeiss software. Additionally, non-DRE voided urine samples collected from clinically determined BPH patients (N = 97), were also analyzed similarly. Results: Urine samples from 62 patients were processed and analyzed. Mean PSA levels by Gleason grade (GG) group were 6.5 ± 4.1 ng/mL for GG1 (N = 10), 7.2 ± 3.8 for GG2 (N = 31), 13.2 ± 14.6 for GG3 (N = 13), 6.2 ± 2.2 for GG4 (N = 2) and 50.2 ± 104.9 for GG5 (N = 6). Like the PSA, % MC shed (66.7 ± 27.7) in voided urine and the fluorescent intensity (35.8 ± 5.7) were highest in patients with GG5 prostate cancer. All PCa patients in GG1 to GG5 shed MC in voided urine with increasing % of MC and increasing fluorescence intensity which correlated with the increasing GG for PCa. For BPH, the specificity for the assay was 89.6% (95% CI:81.9–94.9%), PPV was 0.0% and NPV was 100% (95.9% CI, 95.9–100%). Conclusions: These data indicate the following: (i) PCa MC shed in non-DRE voided urine can be detected by targeting VPAC receptors, (ii) MC are shed in non-DRE voided urine with increasing quantity, corresponding to the severity of the disease, and (iii) this non-DRE voided urine optical assay provides a simple, noninvasive, and reliable method for the preliminary detection of PCa with potentially a lower cost than the currently available pre-biopsy detection technologies. Full article

Background/Objectives: The increasing complexity of spinal oncology procedures, particularly in en-bloc tumor resections, creates challenges in tissue perfusion assessment due to extended operative times and extensive surgical dissection. Real-time visualization of tissue perfusion can be achieved with ICG using commercially available handheld imaging systems, offering potential advantages in spinal oncology cases. This study assessed the utility of ICG in analyzing soft-tissue viability during complex spine procedures extending beyond 7.5 h, with a particular focus on oncologic resections. Methods: Three cases that required over 7.5 h of operative time were chosen for ICG utilization. These cases included an en-bloc malignant peripheral nerve sheath tumor resection, an en-bloc resection of a malignant epithelioid neoplasm, and a long-segment fusion revision for pseudoarthrosis. At the conclusion of the critical portion of the procedure, a handheld intraoperative fluorescence camera was utilized to visualize the tissue penetration of intravenous ICG. Results: Prior to injecting ICG, devascularized tissue was not clearly visible. Injecting ICG allowed clear separation of vascularized (fluorescing) and devascularized (non-fluorescing) tissues. One region of non-florescent tissue was later confirmed to be devascularized with MRI and experienced postoperative infection. Conclusions: As the complexity of spinal oncology procedures increases, ICG fluorescence imaging offers a novel method for real-time assessment of tissue perfusion. This technique may be particularly valuable in extensive tumor resections, post-radiation cases, and revision surgeries where tissue viability is at risk. Further investigation in the spinal oncology population could help establish whether early identification of poorly perfused tissues impacts wound healing outcomes. Full article

Soil remediation for cadmium (Cd) toxicity is essential for successful tobacco cultivation and production. Melatonin application can relieve heavy metal stress and promote plant growth; however, it remains somewhat unclear whether melatonin supplementation can remediate the effects of Cd toxicity on the growth and development of tobacco seedlings. Herein, we evaluated the effect of soil-applied melatonin on Cd accumulation in tobacco seedlings, as well as the responses in growth, physiological and biochemical parameters, and the expression of stress-responsive genes. Our results demonstrate that melatonin application mitigated Cd stress in tobacco, and thus promoted plant growth. It increased root fresh weight, dry weight, shoot fresh weight and dry weight by 58.40%, 163.80%, 34.70% and 84.09%, respectively, compared to the control. Physiological analyses also showed significant differences in photosynthetic rate and pigment formation among the treatments, with the highest improvements recorded for melatonin application. In addition, melatonin application alleviated Cd-induced oxidative damage by reducing MDA content and enhancing the activities of enzymatic antioxidants (CAT, SOD, POD and APX) as well as non-enzymatic antioxidants (GSH and AsA). Moreover, confocal microscopic imaging confirmed the effectiveness of melatonin application in sustaining cell integrity under Cd stress. Scanning Electron Microscopy (SEM) observations illustrated the alleviative role of melatonin on stomata and ultrastructural features under Cd toxicity. The qRT-PCR analysis revealed that melatonin application upregulated the expression of photosynthetic and antioxidant-related genes, including SNtChl, q-NtCSD1, NtPsy2 and QntFSD1, in tobacco leaves. Together, our results suggest that soil-applied melatonin can promote tobacco tolerance to Cd stress by modulating morpho-physiological and biochemical changes, as well as the expression of relevant genes. Full article

This study explored the sustainable valorization of fruit waste extracts from sugarcane bagasse (SB), banana peel (BP), and watermelon rind (WR) for Euglena gracilis biomass and β-1,3-glucan production. The extracts were prepared using water extraction (WE), high-temperature and pressure treatment (HTP), and dilute sulfuric acid treatment (DSA). The DSA-treated extracts consistently yielded the best results. E. gracilis cultured in SB-DSA showed the highest cell density with a 2.08-fold increase compared to the commercial HUT medium, followed by BP-DSA (1.35-fold) and WR-DSA (1.70-fold). Photosynthetic pigment production increased significantly, with chlorophyll a yield being highest in SB-DSA (1.90-fold increase). The chlorophyll a/b ratio and total carotenoid content also improved, indicating enhanced light-harvesting capacity and photoprotection. Photosynthetic efficiency, measured by chlorophyll fluorescence, notably improved. The maximum quantum yield of PSII (Fv/Fm) increased by up to 25.88% in SB-DSA, suggesting reduced stress and improved overall photosynthetic health. The potential photochemical efficiency (Fv/F0) showed even greater improvements: up to 40.53% in SB-DSA. Cell morphology analysis revealed larger cell aspect ratios, implying a more active cellular physiological state. β-1,3-glucan yield also increased by 23.99%, 12.92%, and 23.38% in SB-DSA, BP-DSA, and WR-DSA, respectively. This study demonstrates the potential of pretreated fruit waste as a cost-effective and sustainable medium for E. gracilis cultivation, offering the dual benefits of waste valorization and high-value compound production. These findings contribute to the development of more efficient biorefinery processes and align with the circular economy principles in food biotechnology. Full article

The genetic diversity and structure of 17 populations of J. curcas, including 92 accessions from different provenances (tropical and subtropical), were investigated and effectively evaluated using twelve inter-simple sequence repeats (ISSRs) and seven pairs of florescence-amplified fragment-length polymorphism (AFLP) primers. Genetic diversity, at the overall level among populations of J. curcas based on the ISSR markers, showed that the observed number of alleles (Na) was 1.593, the effective number of alleles (Ne) was 1.330, Nei’s gene diversity (H) was 0.200, Shannon’s information index (I) was 0.303, and the percentage of polymorphic loci was 59.29%, indicating moderate genetic diversity between and within the different populations of J. curcas. Based on the genetic diversity analysis of AFLP markers, there were 1.464 (Na) and 1.216 (Ne) alleles, Nei’s gene diversity (H) was 0.132, Shannon’s information index (I) was 0.204, and the percentage of polymorphic loci was 46.40%. The AMOVA analysis showed that this large variance was due to differences within the populations, with genetic distinctions and limited gene flow among those from varied regions. The 17 populations were clustered into five main groups via UPGMA clustering analysis based on Nei’s genetic distance, and the genetic relationships among the populations exhibited no significant correlations with geographical provenances. The genetic variation among Chinese populations of J. curcas distributed in dry-hot valley areas was remarkable, and the American germplasm presented with distinct genetic differentiation. Full article

Maize is among the most significant crops in the world regarding production and yield, but it is highly sensitive to drought, which reduces the growth, photosynthetic efficiency, grain quality, and yield production of a plant. Quantum yield efficiency of photosystem II is a critical photosynthetic component that is susceptible to drought stress. This study intended to investigate the effects of drought stress on growth and morpho-physiological parameters using three maize hybrids (‘P-3011w’, ‘P-3092’ and ‘iku20’) with contrasting soil moisture contents (100%, 40%) at the pre-flowering stage. The stress treatment (40%) was initiated at stage V7, for a period of 15 days; the experimental units were established in a completely randomized design with split-plot arrangement along with three repetitions in 42 L pots using a substrate of peat moss, black soil and poultry manure (1:2:1). The morphological, growth-related and physiological parameters were assessed, including chlorophyll fluorescence (Fv/Fm), which was measured using a LiCor-6400-40 fluorometer. The results showed that all morphological, growth-related and physiological variables decreased under drought stress during the reproductive stage, with the exception of leaf temperature and intercellular CO₂ concentration, which increased by 12% and 54%, respectively. Drought stress significantly reduced the photosynthetic chlorophyll fluorescence (43%), due to damage to photosystem II. The lowest percentage of damage to photosystem II (34%) was observed in the iku20 genotype. In contrast, P-3011w and P-3092 had the highest levels of significantly similar damage (49% and 46%, respectively). The correlation analysis showed a highly positive interaction of chlorophyll fluorescence (Fv/Fm) with net photosynthetic rate and stomatal conductance under drought conditions, and multiple regression analysis revealed that the maximum effect on net photosynthetic rate under drought was due to the damage it caused to photosystem II. Thus, iku20 might have a tendency to be able to withstand drought stress in the dry northeast region of Mexico. Overall, we concluded that the photosystem II was negatively impacted by drought stress thus causing a reduction in all physiological, morphological and growth-related variables. Full article

Endothelial cells (ECs) maintain vessel tone and barrier integrity, regulate blood homeostasis, and prevent the extravasation of leukocytes under normal physiological conditions. Because of the limited lifespans and batch-to-batch differences with respect to the genetic make-up of primary ECs, established immortal EC lines are extensively used for studying endothelial biology. To address this issue, the immortal endothelial cell line EA.hy926 was developed by fusing primary human umbilical vein endothelial cells (HUVECs) with human lung carcinoma A549 cells. EA.hy926 cells share a number of similar endothelial properties with HUVECs and are considered the immortal counterpart to primary HUVECs. However, the cytogenetic integrity of EA.hy926 cells is not fully elucidated. We characterized EA.hy926 cells with conventional G-banding and molecular cytogenetic techniques such as spectral karyotyping and subtelomeric fluorescence in situ hybridization. Cytogenetic analysis revealed an array of numerical and stable structural chromosomal rearrangements including one deletion, one duplication, one isochromosome, seven simple translocations, and five complex translocations in Ea.hy926 cells. These findings will advance comprehension of EA.hy926 cell biology and augment future endothelial studies, specifically in comparison studies between HUVECs and EA.hy926 cells. Full article

Carbon dots (CDs) have emerged as significant fluorescent nanomaterials due to their bright, stable fluorescence, good biocompatibility, facile synthesis, etc. They are widely used in various scientific and practical applications, particularly in combination with mesoporous, florescent, or magnetic nanomaterials to enhance their properties. Recent research has focused on employing CDs and their composites in drug analysis, drug loading, biological imaging, disease diagnosis, and temperature sensing, with a growing interest in their biological and medical applications. In this study, we synthesized blue-fluorescent S, N-co-doped CDs (cys-CDs) using hydrothermal synthesis with L-cysteine and sodium citrate. These resulting cys-CD particles were approximately 3.8 nm in size and exhibited stable fluorescence with a quantum yield of 0.66. By leveraging the fluorescence quenching of the cys-CDs, we developed a rapid and sensitive method for baicalein detection, achieving high sensitivity in the low micromolar range with a detection limit for baicalein of 33 nM. Our investigation revealed that the fluorescence-quenching mechanism involved static quenching and inner-filter effect components. Overall, cys-CDs proved to be effective for accurate quantitative baicalein detection in real-world samples. Full article

The activation of caspases is a crucial event and an indicator of programmed cell death, also known as apoptosis. These enzymes play a central role in cancer biology and are considered one promising target for current and future advancements in therapeutic interventions. Traditional methods of measuring caspase activity such as antibody-based methods provide fundamental insights into their biological functions, and are considered essential tools in the fields of cell and cancer biology, pharmacology and toxicology, and drug discovery. However, traditional methods, though extensively used, are now recognized as having various shortcomings. In addition, these methods fall short of providing solutions to and matching the needs of the rapid and expansive progress achieved in studying caspases. For these reasons, there has been a continuous improvement in detection methods for caspases and the network of pathways involved in their activation and downstream signaling. Over the past decade, newer methods based on cutting-edge state-of-the-art technologies have been introduced to the biomedical community. These methods enable both the temporal and spatial monitoring of the activity of caspases and their downstream substrates, and with enhanced accuracy and precision. These include fluorescent-labeled inhibitors (FLIs) for live imaging, single-cell live imaging, fluorescence resonance energy transfer (FRET) sensors, and activatable multifunctional probes for in vivo imaging. Recently, the recruitment of mass spectrometry (MS) techniques in the investigation of these enzymes expanded the repertoire of tools available for the identification and quantification of caspase substrates, cleavage products, and post-translational modifications in addition to unveiling the complex regulatory networks implicated. Collectively, these methods are enabling researchers to unravel much of the complex cellular processes involved in apoptosis, and are helping generate a clearer and comprehensive understanding of caspase-mediated proteolysis during apoptosis. Herein, we provide a comprehensive review of various assays and detection methods as they have evolved over the years, so to encourage further exploration of these enzymes, which should have direct implications for the advancement of therapeutics for cancer and other diseases. Full article

Canna glauca, an ornamental plant widely cultivated in aquatic habitats, is notable for its long florescence and showy flowers. The flower of this species is distinguished by its petaloid staminodes, which comprise the majority of the overall floral display. Flavonoids have been reported to be the predominant pigment groups that determine most flower colors. However, the influence of flavonoid metabolic pathways on the flower color of C. glauca remains to be investigated. In this study, comprehensive floral transcriptomes and metabolite profiles of the wild type (yellow flower) and ‘Erebus’ cultivar (pink flower) of C. glauca were analyzed. We identified 432 flavonoid metabolites, including 20 anthocyanins. ‘Erebus’ accumulated higher levels of 18 anthocyanins than the wild type, including 10 cyanidins, 4 pelargonidins, and 4 peonidins. The wild type accumulated higher levels of two malvidins. Through the joint analysis of transcriptomics and metabonomics, we observed a notable association between the expression of three DEGs and eleven anthocyanin levels. Furthermore, we analyzed the expression patterns of key genes that determine flavonoid biosynthesis, such as CHS, CHI, F3′H, and DFR. These findings provide enlightenment on the anthocyanin accumulation of Canna glauca, serving as a basis for exploring biochemical and molecular mechanisms underlying flower coloration. Full article

Industrial hemp Cannabis sativa L. is an economically important crop mostly grown for its fiber, oil, and seeds. Due to its increasing applications in the pharmaceutical industry and a lack of knowledge of gene functions in cannabinoid biosynthesis pathways, developing an efficient transformation platform for the genetic engineering of industrial hemp has become necessary to enable functional genomic and industrial application studies. A critical step in the development of Agrobacterium tumefaciens-mediated transformation in the hemp genus is the establishment of optimal conditions for T-DNA gene delivery into different explants from which whole plantlets can be regenerated. As a first step in the development of a successful Agrobacterium tumefaciens-mediated transformation method for hemp gene editing, the factors influencing the successful T-DNA integration and expression (as measured by transient β-glucuronidase (GUS) and Green Florescent Protein (GFP) expression) were investigated. In this study, the parameters for an agroinfiltration system in hemp, which applies to the stable transformation method, were optimized. In the present study, we tested different explants, such as 1- to 3-week-old leaves, cotyledons, hypocotyls, root segments, nodal parts, and 2- to 3-week-old leaf-derived calli. We observed that the 3-week-old leaves were the best explant for transient gene expression. Fully expanded 2- to 3-week-old leaf explants, in combination with 30 min of immersion time, 60 µM silver nitrate, 0.5 µM calcium chloride, 150 µM natural phenolic compound acetosyringone, and a bacterial density of OD_600nm = 0.4 resulted in the highest GUS and GFP expression. The improved method of genetic transformation established in the present study will be useful for the introduction of foreign genes of interest, using the latest technologies such as genome editing, and studying gene functions that regulate secondary metabolites in hemp. Full article

Streptococcus mutans, the primary cause of dental caries, relies on its ability to create and sustain a biofilm (dental plaque) for survival and pathogenicity in the oral cavity. This study was focused on the antimicrobial biofilm formation control and biofilm dispersal potential of Coumaric acid (CA) against Streptococcus mutans on the dentin surface. The biofilm was analyzed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) viability assay, microtiter plate assay, production of extracellular polymeric substances (EPSs), florescence microscopy (surface coverage and biomass μm²) and three-dimensional (3D) surface plots. It was observed that CA at 0.01 mg/mL reduced bacterial growth by 5.51%, whereases at 1 mg/mL, a significant (p < 0.05) reduction (98.37%) was observed. However, at 1 mg/mL of CA, a 95.48% biofilm formation reduction was achieved, while a 73.45% biofilm dispersal (after 24 h. treatment) was achieved against the preformed biofilm. The MTT assay showed that at 1 mg/mL of CA, the viability of bacteria in the biofilm was markedly (p < 0.05) reduced to 73.44%. Moreover, polysaccharide (EPS) was reduced to 24.80 μg/mL and protein (EPS) to 41.47 μg/mL. ImageJ software (version 1.54 g) was used to process florescence images, and it was observed that the biofilm mass was reduced to 213 (μm²); the surface coverage was reduced to 0.079%. Furthermore, the 3D surface plots showed that the untreated biofilm was highly dense, with more fibril-like projections. Additionally, molecular docking predicted a possible interaction pattern of CA (ligand) with the receptor Competence Stimulating Peptide (UA159sp, PDB ID: 2I2J). Our findings suggest that CA has antibacterial and biofilm control efficacy against S. mutans associated with dental plaque under tested conditions. Full article

Alzheimer’s disease (AD) is the most widespread form of senile dementia worldwide and represents a leading socioeconomic problem in healthcare. Although it is widely debated, the aggregation of the amyloid β peptide (Aβ) is linked to the onset and progression of this neurodegenerative disease. Molecules capable of interfering with specific steps in the fibrillation process remain of pharmacological interest. To identify such compounds, we have set up a small molecule screening process combining multiple experimental methods (UV and florescence spectrometry, ITC, and ATR-FTIR) to identify and characterise potential modulators of Aβ_1-42 fibrillation through the description of the biochemical interactions (molecule–membrane Aβ peptide). Three known modulators, namely bexarotene, Chicago sky blue and indomethacin, have been evaluated through this process, and their modulation mechanism in the presence of a biomembrane has been described. Such a well-adapted physico-chemical approach to drug discovery proves to be an undeniable asset for the rapid characterisation of compounds of therapeutic interest for Alzheimer’s disease. This strategy could be adapted and transposed to search for modulators of other amyloids such as tau protein. Full article

Thrips have become some of the most challenging pests to control in mango production due to their short developmental time, hidden locations and resistance to pesticides, in the tropical regions of China. To improve pesticides efficacy, the tank-mix adjuvants Qi Gong (QG) and the thrips attractant Lv Dian (LD) were added when using an unmanned aerial vehicle (UAV) to control thrips. The surface tension, contact angle on mango leaves, droplet size, spreading rate, and drying time of the two tank-mix adjuvants were determined. The effects of the two tank-mix adjuvants using a UAV sprayer on the droplet coverage and control efficacy against thrips on mango inflorescences were tested through field trials. The results showed that both QG and LD could reduce the surface tension of the liquid and the contact angle on mango leaves and could increase the spreading performance. The droplet coverage in the upper layer of the canopy was about 2% higher than that in the lower layer, and the coverage at the top of the panicle was 5% higher than that at the bottom. QG improved the deposition coverage of mango inflorescences by about 31.5%. The addition of QG increased the efficacy by 18.24% and 8.03%, respectively, at florescence and the young fruit stage. The addition of the LD increased the efficacy by 24.56% and 14.38%, respectively, at florescence and the young fruit stage. These test results can provide a scientific basis for the control of mango thrips with UAVs. Full article