Search Results (184)

Mabroom dates (Phoenix dactylifera L.) are recognized as one of the most important crops in Qatar. Fresh fruit dates are susceptible to mould and post-harvest spoilage, resulting in a significant financial loss. Octanoic fatty acid (OFA) has been shown to regulate the growth of mould-causing organisms such as fungi and bacteria. It is known to have antibacterial properties. The objective of the current study was to evaluate the in vitro effect of OFA on the post-harvest pathogens of Mabroom fruits. Fresh, apparently healthy, and fully ripe Mabroom dates were obtained from the National Agriculture and Food Corporation (NAFCO). The chosen fruits were packed in sterile, well-ventilated plastic boxes and transported to the lab under controlled conditions. The fruits were distributed into five groups (G1 to G5). The groups G1, G2, and G3 received 1%, 2%, and 3.5% OFA, respectively, while G4 was left untreated and G5 was washed only with tap water as a positive control treatment. Each group contained 200 g of fresh and healthy semi-soft dates. The samples were then dried and incubated in a humidity chamber at 25 °C ± 2 for seven days. The signs and symptoms of decay were monitored and recorded. The presence of pathogens was confirmed via phenotypic and microscopic-based methods. The results showed a significant difference (p ≤ 0.05) among the groups. OFA at 3.5% had the strongest inhibitory action against post-harvest pathogens, followed by OFA2%. However, there were no differences (p ≤ 0.05) between OFA1% and the control groups. Aspergillus spp., Penicillium spp., Rhizopus spp., and Botrytis spp. were most abundant in the control group, followed by OFA2% and OFA1%, respectively. In conclusion, octanoic fatty acid at 3.5% may improve the quality of date fruits through its high antimicrobial activity, reduce the effect of post-harvest decay, minimize the loss of date fruits during storage, and improve the sustainability of date fruits. Further experiments are necessary to confirm the effectiveness of OFA as a green solution for sustainable date fruit production. Full article

Maize (Zea mays L.), a vital global food crop, relies on its stomatal structure for regulating photosynthesis and responding to drought. Conventional manual stomatal detection methods are inefficient, subjective, and inadequate for high-throughput plant phenotyping research. To address this, we curated a dataset of over 1500 maize leaf epidermal stomata images and developed a novel lightweight detection model, StomaYOLO, tailored for small stomatal targets and subtle features in microscopic images. Leveraging the YOLOv11 framework, StomaYOLO integrates the Small Object Detection layer P2, the dynamic convolution module, and exploits large-scale epidermal cell features to enhance stomatal recognition through auxiliary training. Our model achieved a remarkable 91.8% mean average precision (mAP) and 98.5% precision, surpassing numerous mainstream detection models while maintaining computational efficiency. Ablation and comparative analyses demonstrated that the Small Object Detection layer, dynamic convolutional module, multi-task training, and knowledge distillation strategies substantially enhanced detection performance. Integrating all four strategies yielded a nearly 9% mAP improvement over the baseline model, with computational complexity under 8.4 GFLOPS. Our findings underscore the superior detection capabilities of StomaYOLO compared to existing methods, offering a cost-effective solution that is suitable for practical implementation. This study presents a valuable tool for maize stomatal phenotyping, supporting crop breeding and smart agriculture advancements. Full article

Kernicterus spectrum disorder is the permanent and highly disabling neurologic sequel of neonatal exposure to hyperbilirubinemia, presenting, among other symptoms, variable and untreatable motor disabilities. To search for potential biomolecular explanations, we used a Gunn rat colony exhibiting spontaneous hyperbilirubinemia and a large variability of motor deficits on a beam-walking test. Histological and microscopic analyses confirmed worsening damage in the cerebellum (Cll; hypoplasia, increased death of neurons, and disrupted astroglial structures) and parietal motor cortex (hCtx; increased cell sufferance and astrogliosis). Clustering and network analyses of transcriptomic data reveal rearrangement of the physiological expression patterns and signaling pathways associated with bilirubin neurotoxicity. Bilirubin content among hyperbilirubinemic (jj) animals is overlapped, which suggests that the amount of bilirubin challenge does not fully explain the tissue, transcriptomic, proteomic, and neurobehavioral alterations. The expression of nine genes involved in key postnatal brain development processes is permanently altered in a phenotype-dependent manner. Among them, Grm1, a metabotropic glutamatergic receptor involved in glutamate neurotoxicity, is consistently downregulated in both brain regions both at the transcriptomic and proteomic levels. Our results support the role of Grm1 and glutamate as biomolecular markers of ongoing bilirubin neurotoxicity, suggesting the possibility to improve diagnosis by ¹H-MR spectroscopy. Full article

Collagen II is a vital structural component in developing bones and mature cartilage. Mutations in this protein cause spondyloepiphyseal dysplasia, a disease characterized primarily by altered skeletal growth and manifesting with a range of phenotypes, from lethal to mild. This study examined transgenic mice harboring the R992C (p.R1124C) substitution in collagen II. Previous research demonstrated significant growth abnormalities and disorganized growth plate structure in these mice, and histological signs of osteoarthritic changes in the knee joints of 9-month-old mice with the R992C mutation. Our study focuses on detecting early structural changes in the articular cartilage that occur before histological signs become apparent. Through microscopic and spectroscopic analyses, we observed significant alterations in the distribution gradients of collagenous proteins and proteoglycans in the cartilage of R992C mutant mice. We propose that these early changes, eventually leading to articular cartilage degeneration in older mice, underscore the progressive nature of osteoarthritic changes linked to collagen II mutations. By identifying these early structural aberrations, our findings emphasize the importance of early detection of osteoarthritic changes, potentially facilitating timely, non-surgical interventions. Full article

Objectives: The oral cavity hosts one of the most complex microbial communities in the body. A disruption of the balance favors the growth of pathogenic species, contributing to oral diseases. The rise in microbial resistance has limited the effectiveness of conventional treatments, shifting the interest to natural product-based alternatives. Given its superior bioavailability and bioactivity in other models, this study investigates the antifungal potential of a novel curcumin derivative, PAC (3,5-bis(4-hydroxy-3-methoxybenzylidene)-N-methyl-4-piperidone), and studies its impact on host–pathogen dynamics and host defense mechanisms. Methods: Candida albicans was used as the model organism. Viability, growth kinetics, and colony formation were evaluated using optical density, agar culture, and MTT assay. Biofilm formation was assessed through electron microscopy and total sugar quantification. The morphological transition from hyphae to the less virulent blastospore was monitored using an optical microscope. The gene expression of adhesion factors and host defense markers was analyzed using RT-PCR. Results: PAC impairs C. albicans viability and reduces virulence by compromising biofilm formation and ensuring phenotypic transition to a blastospore form. Also, PAC controls C. albicans growth via necrosis/ROS pathways. As a result, PAC appears to repress host–pathogen interaction by downregulating SAPs, EAP1, and HWP1 adhesion genes, thus relieving the need to activate gingival epithelial cell defense mechanisms. This is highlighted by recording baseline levels of IL-6, IL-8, and IL-1β cytokines and antimicrobial β-defensin peptides in the presence of less virulent candida forms. Conclusions: PAC effectively reduces C. albicans virulence by limiting biofilm formation and adhesion while minimizing inflammatory responses. These findings support its potential as a promising therapeutic agent for infectious disease control. Full article

This paper aims to present the phenotypic characteristics, such as length, width, circular diameter, volume, surface area to cross-sectional area ratio, surface uniformity, and surface texture, of the foodborne pathogens Salmonella enterica serovar Agona, Salmonella enterica serovar Enteritidis, Listeria monocytogenes, and Campylobacter jejuni. It is a novel “field-guide” presentation of the observable morphological characteristics of these four species in the form of a Dichotomous Key. Phenotypic values of the pathogens were measured using a high-accuracy, nano-level-resolution 3D laser confocal scanning microscope. All samples were fixed with glutaraldehyde and stained with methylene blue, ensuring morphological preservation. The images of the pathogens were captured under various imaging modes, including 3D, laser, and transmission electron microscopy resolutions. Results show that these pathogens exhibit distinct morphological and surface properties, with Campylobacter jejuni showing unique features. The dichotomous key shows a simplified way of classifying selected foodborne pathogens from their morphometric properties. The morphometric measurements, in many instances first reported in the literature, and images provide a readily observable way to identify and classify microorganisms, allowing researchers to potentially study evolutionary relationships, assess species diversity, and understand how organisms interact with their environment, especially when genetic information is limited or difficult to obtain. Full article

In homeostasis, the glial cells support pivotal functions, such as neuronal differentiation, neuroprotection, nutrition, drug metabolism, and immune response in the central nervous system (CNS). Among these cells, astrocytes and microglia have been highlighted due to their role in the pathogenesis of several diseases or due to their role in the defense against several insults (ex., chemicals, and pathogens). In Vitro cytological analysis of astrocytes and microglia has contributed to the understanding of the role of morphological changes in glial cells associated with a neuroprotective or neurotoxic phenotype. Currently, the main tools used for the investigation of glial cell morphology in culture are phase contrast microscopy or immunolabeling/fluorescence microscopy. However, generally, phase contrast microscopy does not generate images with high resolution and therefore does not contribute to visualizing a single cell morphology in confluent cell cultures. On the other hand, immunolabeling requires high-cost consumable antibodies, epifluorescence microscope or confocal microscope, and presents critical steps during the procedure. Therefore, identifying a fast, reproducible, low-cost alternative method that allows the evaluation of glial morphology is essential, especially for neuroscientists from low-income countries. This article aims to revise the use of Rosenfeld’s staining, as an alternative low-cost and easy-to-reproduce method to analyze astrocytic and microglial morphology in culture. Additionally, it shows Rosenfeld’s staining as a valuable tool to analyze changes in neural cell morphology in toxicological studies. Full article

Seed rope direct-seeding is an advanced precision sowing technique that involves encapsulating seeds within rope materials, adhering to specific spacing and quantity, and then deploying these ropes in the field as an alternative to conventional direct-seeding. This method offers the dual benefits of minimal sprout damage and precise control over row-to-seed spacing. The mechanical properties of the seed rope material and the integrity of the wrapped seeds are critical factors that influence the growth and development of the plants’ root system, which in turn is a key determinant for the optimization of the seed rope automatic seeder. This paper employed uniaxial tensile testing to investigate the mechanical properties and tensile failure characteristics of seed ropes across various materials, seed wrapping techniques, and seed soaking methods. Additionally, scanning electron microscopy was utilized to scrutinize the microstructural features of the tensile fracture surfaces of the seed ropes. The results showed that the tensile strength of paper-based seed ropes ranged from 1.80 to 2.89 N/mm, with elongation at the break between 31.4% and 47.5%, and a critical stress range of 5.67 to 9.06 N. In contrast, non-woven fabric ropes exhibited a tensile strength range of 0.91 to 1.23 N/mm², an elongation at break range of 160.3 to 284.2%, and a critical stress range of 2.86 to 3.86 N. Electron microscope scanning imagery analysis indicated that the broken fibers were disordered, and the fibers of the soaked ropes showed minor surface damage, which is attributed to the decline in tensile strength observed in soaked ropes. Regarding the phenotypic study of root growth and development, the root growth and development phenotypes of two types of rope materials across four different vegetable varieties were explored; the results indicated that the influence of the seed rope material on the root system was pronounced in the early stages of growth and development. As plants progressed to the middle stage of growth, the trend in root length mirrored that of the early stage, with the seed rope material continuing to significantly impact root system development. In the late stage of growth, the effect of the seed rope material on root growth gradually diminished as the seed rope material decomposed. Interestingly, the root length under non-woven fabric wrapping not only caught up to, but in some instances, surpassed the root length of unwrapped seeds. This research provides valuable theoretical insights and data to support the optimization of the parameters for the automatic seed rope direct-seeder. Full article

Several etiologic factors for the development of acute leukemias have been suggested; however, none is applicable to all cases. We isolated a certain mycovirus-containing Aspergillus flavus (MCAF) from the home of a patient with acute lymphoblastic leukemia. Repeated electron microscopic evaluations proved the existence of mycovirus in this organism. According to chemical analysis, this organism does not produce any aflatoxin, possibly due to its infestation with mycoviruses. We reported that using the ELISA technique, forty pediatric patients with acute lymphoblastic leukemia (ALL) uniformly had antibodies to the products of MCAF. In contrast, three separate groups of controls, consisting of normal blood donors, individuals with solid tumors, and patients with sickle cell disease, were negative. In vitro exposure of mononuclear blood cells from patients with ALL, in full remission, to the products of MCAF induced redevelopment of cell surface phenotypes and genetic markers characteristic of ALL. The controls were negative. The incubation of normal and ALL cell lines with the products of MCAF resulted in significant cellular apoptosis, changes in the cell cycle, and the downregulation of transcription factors, including PAX-5 and Ikaros (75 and 55 kDa). Fungi are widespread in nature, and many contain mycoviruses. Normally, an individual inhales 1 to 10 fungal spores per minute, while farmers can inhale up to 75,000 spores per minute. It is known that farmers and foresters, who are more exposed to fungi, have a higher rate of acute leukemia. In contrast, asthmatics, most of whom are allergic to fungal agents, and individuals working in office settings have a lower rate. One of the theories for the development of acute leukemia suggests a genetic predisposition followed by exposure to an infectious agent. With the above findings, we propose that mycovirus-containing Aspergillus flavus may have an etiological role in leukemogenesis in immune-depressed and genetically susceptible individuals. Full article

Background: The crosstalk between cancer-associated fibroblasts (CAFs) and tumor cells promotes proliferation, tumor relapse, and the acquisition of a partial epithelial-to-mesenchymal (pEMT) phenotype in tumor cells. The aim of this study was to investigate the effects of patient-derived CAFs on tumor cell growth and radioresistance in head and neck squamous cell carcinoma (HNSCC). Methods: CAFs were isolated and cultured in a three-dimensional spheroid formation. SCC-25 tumor cells educated by the CAFs (SCC25-E cells) were subjected to irradiation, and the response of the CAF-stimulated tumor cells to radiotherapy was determined using an MTT assay, a clonogenic assay, and Western blotting. Tumor cell morphological changes and growth dynamics were assessed using 3D holotomographic microscopy and a live video microscope. Results: Patient-derived CAFs significantly increased the growth rate of SCC-25 cells. CAFs drove fibrosis in the tumor microenvironment (TME), functioned as a physical barrier, temporarily stopped tumor growth, and induced the p38 mitogen-activated protein kinase (MAPK) signaling pathway. Viability after irradiation at 4–8 Gy was significantly higher in SCC25-E cells than in the controls (p = 8 × 10^–4 or lower). Furthermore, irradiation triggered the pEMT profile in HNSCC cells. Conclusions: CAFs’ education of tumor cells and the induced p38 phosphorylation had no influence on irradiation sensitivity. SCC25-E cultures demonstrated increased tumor cell growth, viability, and stress-induced phospho-p38 activation. Full article

Mung bean (Vigna radiata) plays a significant role in agricultural trade, food processing and utilization, and cropping structure adjustment due to its abundant nutritional components, medicine-food homology, capacity for nitrogen fixation, and soil improvement. The low yield level is a crucial limitation factor in the mung bean industry, while heterosis is an efficient path for increasing crop yields. The flexible utilization of male sterile mung bean materials may solve this pressing demand in the industry. This study identified a completely male-sterile mutant, vrnp 1, in the EMS-mutagenized mung bean cultivar Jilv 10 population, which is controlled by a single recessive nuclear gene. Furthermore, we employed a series of microscopical and histological techniques and observed that the tapetal cells in the vrnp 1 mutant did not perform as expected when reaching stage 8 of anther development, notably exhibiting a delay in entering PCD. This was accompanied by a failure to deposit cell wall components onto the pollen wall, culminating in a complete absence of mature pollen and the manifestation of male sterility. In conclusion, the vrnp 1 mutant could potentially serve as a promising candidate for male sterility in exploiting hybrid vigor in mung bean. Our research may elucidate how the delayed initiation of programmed cell death in tapetal cells contributes to a factor implicated in mung bean male sterility. Furthermore, the phenotypic data collected during pivotal developmental phases may have contributed to a better grasp of mung bean microspores and anther development. Full article

Spontaneous abortion, commonly known as miscarriage, is a significant concern during early pregnancy. Histopathological examination of tissue samples is a widely used method to diagnose and classify tissue phenotypes found in products of conception (POC) after spontaneous abortion. Background: Histopathological examination is subjective and dependent on the skill and experience of the examiner. In recent years, artificial intelligence (AI)-based techniques have emerged as a promising tool in medical imaging, offering the potential to revolutionize tissue phenotyping and improve the accuracy and reliability of the histopathological examination process. The goal of this study was to investigate the use of AI techniques for the detection of various tissue phenotypes in POC after spontaneous abortion and evaluate the accuracy and reliability of these techniques compared to traditional manual methods. Methods: We present a novel publicly available dataset named HistoPoC, which is believed to be the first of its kind, focusing on spontaneous abortion (miscarriage) in early pregnancy. A diverse dataset of 5666 annotated images was prepared from previously diagnosed cases of POC from Atia General Hospital, Karachi, Pakistan, for this purpose. The digital images were prepared at 10× through a camera-connected microscope by a consultant histopathologist. Results: The dataset’s effectiveness was validated using several deep learning-based models, demonstrating its applicability and supporting its use in intelligent diagnostic systems. Conclusions: The insights gained from this study could illuminate the causes of spontaneous abortion and guide the development of novel treatments. Additionally, this study could contribute to advancements in the field of tissue phenotyping and the wider application of deep learning techniques in medical diagnostics and treatment. Full article

Background/Objectives: This study aims to investigate the effects of 4-methylumbelliferone (4-MU) on islet morphology, cell phenotype and function, and to explore possible mechanisms of β cell regeneration. Methods: The Type 1 diabetes (T1D) model was induced by continuous dose injection of streptozotocin (STZ), and mice were treated with 4-MU for 3 weeks. Plasma insulin level, islet cell phenotype and immune infiltration were determined by IPGTT, ELISA, HE and immunofluorescence. The Ins2^Cre/+/Rosa26-eGFP transgenic mice model was used to detect β identity change. Primary rodent islets were incubated with 4-MU or vehicle in the presence or absence of STZ, AO/PI staining, and a scanning electron microscope (SEM), PCR and ELISA were used to evaluated islet viability, islet morphology, the specific markers of islet β cells and insulin secretion. Results: Treatment with 4-MU significantly decreased blood glucose and increased plasma insulin levels in STZ-induced diabetes. The plasma insulin level in the STZ group was 7.211 ± 2.602 ng/mL, which was significantly lower than the control group level (26.94 ± 4.300 ng/mL, p < 0.001). In contrast, the plasma insulin level in the STZ + 4-MU group was 22.29 ± 7.791 ng/mL, which was significantly higher than the STZ group (p < 0.05). The 4-MU treatment increased islet and β cells numbers and decreased α cell numbers in STZ-induced diabetes. Conclusions: Islet inflammation as indicated by insulin and CD3 was caused by infiltrates, and the β cell proliferation as indicated by insulin and Ki67 was boosted by 4-MU. β cell dedifferentiation was inhibited by 4-MU as assessed by insulin and glucagon double-positive cells and confirmed by Ins2^Cre/+/Rosa26-eGFP mice. In cultured primary rodent islets, 4-MU restored islet viability, protected islet morphology, inhibited β-cell dedifferentiation, and promoted insulin secretion. The benefits of 4-MU in T1D have been proved to be associated with β cells self-replication, dedifferentiation inhibition and immune progression suppression, which help to maintain β cell mass. Full article

MYBPC3, encoding cardiac myosin binding protein-C (cMyBP-C), is the most mutated gene known to cause hypertrophic cardiomyopathy (HCM). However, since little is known about the underlying etiology, additional in vitro studies are crucial to defining the underlying molecular mechanisms. Accordingly, this study aimed to investigate the molecular mechanisms underlying the pathogenesis of HCM associated with a polymorphic variant (D389V) in MYBPC3 by using isogenic human-induced pluripotent stem cell (hiPSC)-derived cardiac organoids (hCOs). The hiPSC-derived cardiomyocytes (hiPSC-CMs) and hCOs were generated from human subjects to define the molecular, cellular, functional, and energetic changes caused by the MYBPC3^D389V variant, which is associated with increased fractional shortening and highly prevalent in South Asian descendants. Recombinant C0-C2, N’ region of cMyBP-C (wild-type and D389V), and myosin S2 proteins were also utilized to perform binding and motility assays in vitro. Confocal and electron microscopic analyses of hCOs generated from noncarriers (NC) and carriers of the MYBPC3^D389V variant revealed the presence of highly organized sarcomeres. Furthermore, functional experiments showed hypercontractility, faster calcium cycling, and faster contractile kinetics in hCOs expressing MYBPC3^D389V than NC hCOs. Interestingly, significantly increased cMyBP-C phosphorylation in MYBPC3^D389V hCOs was observed, but without changes in total protein levels, in addition to higher oxidative stress and lower mitochondrial membrane potential (ΔΨm). Next, spatial mapping revealed the presence of endothelial cells, fibroblasts, macrophages, immune cells, and cardiomyocytes in the hCOs. The hypercontractile function was significantly improved after the treatment of the myosin inhibitor mavacamten (CAMZYOS^®) in MYBPC3^D389V hCOs. Lastly, various vitro binding assays revealed a significant loss of affinity in the presence of MYBPC3^D389V with myosin S2 region as a likely mechanism for hypercontraction. Conceptually, we showed the feasibility of assessing the functional and molecular mechanisms of HCM using highly translatable hCOs through pragmatic experiments that led to determining the MYBPC3^D389V hypercontractile phenotype, which was rescued by the administration of a myosin inhibitor. Full article

Stomata regulate CO₂ and water vapor exchange between leaves and the atmosphere, serving as a vital indicator of climate change resilience. Therefore, understanding the difference in stomatal numbers and patterns among different soybean cultivars across growth stages is essential to comprehending the complex mechanisms underlying soybean adaptation to climate change. The accurate measurements of stomatal density in soybean leaves are essential to understanding the complexity of stomatal density by environmental conditions. We demonstrated that the five epidermal sections and five microscopic images taken from both sides of each epidermal section at each leaf position (tip, middle, and bottom) were sufficient for stomatal measurements. Furthermore, we investigated variations in stomatal density among leaflet locations (left, right, and central) and leaf position across different growth stages. Notably, while there was no significant variation between the two leaves of the vegetative cotyledon (VC) stage and among the three leaflets of the V1 (first trifoliate) to V4 (fourth trifoliate) growth stages, leaves of the VC stage exhibited the lowest stomatal density, whereas those of the V4 stage exhibited the highest stomatal density. These findings could serve as a valuable tool for evaluating stomatal density, analyzing physiological differences under adverse climatic conditions, and phenotyping a large-scale population to identify the genetic factors responsible for stomatal density variations in soybean genotypes. Full article