Search Results (194)

Cerebral palsy (CP) is a group of permanent, but not immutable, disorders of movement and posture caused by a non-progressive lesion or an abnormality in the development of the immature brain. It is the leading cause of motor disability in childhood. The most common sign is muscle spasticity, which leads to the development of contractures and bone deformities. Among the treatments for spasticity, botulinum toxin injection has the strongest scientific evidence and is used from the age of 2 years. In this article, we report three pediatric patients with CP who developed muscle fasciculations after botulinum toxin injection, which were administered according to planned dosages and required clinical monitoring and stretching by a healthcare professional. We review the literature and analyze the possible physiopathological mechanisms; however no other similar clinical cases have been reported. In conclusion, we propose to include muscle fasciculations as a potential transient and non-harmful adverse effect of botulinum toxin infiltration, which remains the treatment of choice for muscle spasticity when combined with rehabilitation in cerebral palsy. Full article

A three-year-old male bearded dragon (Pogona vitticeps) exhibited acute anorexia. Biochemistry revealed mild hyperproteinemia (88 g/L) and elevated liver enzymes (ALT 60 U/L, AST 272 U/L), while the hematology report showed marked lymphocytosis. The animal had been clinically normal at a routine examination 10 months earlier. Based on the clinical and laboratory findings, acute lymphoblastic leukemia was suspected. Treatment was initiated with methylprednisolone (1 mg/kg PO q24h), marbofloxacin (10 mg/kg IM q24h), and lomustine (80 mg/m² PO q14d), calculated according to reptile-specific body surface area formulas. A transient stabilization was followed by sudden deterioration on day 3, characterized by hematemesis and severe respiratory distress, leading to spontaneous death. A complete necropsy including histopathology and anti-CD3 immunohistochemistry revealed disseminated infiltration of neoplastic T-lymphocytes throughout all major visceral organs and confirmed the diagnosis of T-cell lymphoblastic lymphoma/leukemia (L/L). This case represents a rare report of systemic acute lymphoblastic L/L in a bearded dragon and underlies the importance of comprehensive diagnostics in reptiles with non-specific clinical signs and the challenges in the treatment of neoplastic diseases in exotic species. Full article

Chimeric antigen receptor (CAR) T-cell infusion has led to improved outcomes in patients with B-lymphoblastic leukemia, B-cell lymphoma, and multiple myeloma. The spectrum of post-CAR T-cell hematolymphoid abnormalities is expanding, although they remain under-recognized. Pathologists play a key role in characterizing hematolymphoid proliferation after CAR T-cell therapy. This review presents clinical and pathologic findings of common hematolymphoid proliferation after CAR T-cell therapy, illustrated by selected cases. A review of the literature is presented in the context of individual cases, and our current understanding of the pathomechanism is discussed. Infused CAR T-cells undergo a series of four phases: distribution, expansion, contraction, and persistence. In the expansion phase, transient peripheral blood lymphocytosis occurs, reaching a peak two weeks post-infusion. Delayed contraction of CAR T-cells may give rise to hemophagocytic lymphohistiocytosis-like syndrome. Immune effector cell-associated enterocolitis presents in the persistence phase, about 3–6 months after infusion. Pathologic findings include a T-cell infiltrate in the intestinal mucosa and changes resembling graft versus host disease (GVHD). This entity requires differentiation from infections and from T-cell neoplasms, including those derived from CAR T-cells. Secondary myeloid malignancies follow the same pathways as therapy-related myeloid neoplasm but present with a shorter median latency. It is essential for pathologists to recognize post-CAR T-cell hematolymphoid proliferation to support clinical decision making in a high-risk patient population. Full article

Monocytes/macrophages promote the repair of acutely injured muscle while contributing to dystrophic changes in chronically injured muscle in Duchenne muscular dystrophy (DMD) patients and animal models including mdx and mdx^5cv mice. To elucidate the molecular mechanisms underlying this functional difference, we compared the transcriptomes of intramuscular monocytes/macrophages from wild-typed (WT) uninjured muscles, WT acutely injured muscles, and mdx^5cv dystrophic muscles, using single cell-based RNA sequencing (scRNA-seq) analysis. Our study identified multiple transcriptomically diverse monocyte/macrophage subclusters, which appear to be induced by the intramuscular microenvironment. They expressed feature genes differentially involved in muscle inflammation, regeneration, and extracellular matrix (ECM) remodeling, but none of them conform to strict M1 or M2 activation. The Gpnmb⁺Spp1⁺ macrophage subcluster, an injury-associated subcluster that features the signature genes of reported scar-associated macrophages (SAMs) involved in ECM remodeling and fibrosis, is present transiently in acutely injured muscle and persistently in chronically injured dystrophic muscle, along with the persistence of monocytes. Our findings suggest that the persistent monocyte/macrophage infiltration and activation induced by continuous injury may underlie the pathogenic roles of macrophages in mdx^5cv muscles. Controlling muscle injury and subsequent macrophage infiltration and activation may be important to the treatment of DMD. Full article

Sunflower (Helianthus annuus L.) is an important oilseed crop in Northwest China, exhibiting resistance to salt and drought. Mining its excellent tolerance genes can be used for breeding. However, the current platforms for identifying gene function in sunflower is inadequate. The transient transformation system, which can rapidly validate gene function, shows promising prospects in research. In this study, we established an efficient transient expression transformation system for sunflower using three methods: Agrobacterium-mediated infiltration, injection, and ultrasonic-vacuum. The detailed procedures were as follows: Agrobacterium GV3101 carrying a GUS reporter gene on the pBI121 vector with an OD₆₀₀ of 0.8 as the bacterial suspension and 0.02% Silwet L-77 as the surfactant were utilized in all three approaches. For the infiltration method, seedlings grown hydroponically for 3 days were immersed in a bacterial suspension containing 0.02% Silwet L-77 for 2 h; for the injection method, the same solution was injected into the cotyledons of seedlings grown in soil for 4 to 6 days. Subsequently, the seedlings were cultured in the dark at room temperature for three days; for the ultrasonic-vacuum method, seedlings cultured in Petri dishes for 3 days were first subjected to ultrasonication at 40 kHz for 1 min, followed by vacuum infiltration at 0.05 kPa for 5–10 min. Agrobacterium-mediated transient transformation efficiency achieved by the three methods exceeded 90%, with gene expression being sustained for at least 6 days. Next, we employed the infiltration-based sunflower transient transformation technology with the Arabidopsis stable transformation platform to confirm salt and drought stress tolerance of candidate gene HaNAC76 from sunflower responding to various abiotic stresses. Altogether, this study successfully established an Agrobacterium-mediated transient transformation system for sunflower using these three methods, which can rapidly identify gene function and explore the molecular mechanisms underlying sunflower’s resistance traits. Full article

Rainfall infiltration is a key factor affecting the stability of the slope. To study the impact of rainfall on the instability mechanism and stability of slopes, this paper employs numerical simulation to establish a rainfall infiltration slope model and conducts a saturated–unsaturated slope flow and solid coupling numerical analysis. By combining the strength reduction method with the calculation of slope stability under rainfall infiltration, the safety factor of the slope is obtained. A comprehensive analysis is conducted from the perspectives of the seepage field, displacement field and other factors to examine the impact of heavy rainfall patterns and rainfall intensities on the instability mechanism and stability of the slope. The results indicate that heavy rainfall causes the transient saturation zone within the landslide body to continuously move upward, forming a continuous sliding surface inside the slope, which may lead to instability and sliding of the soil in the upper part of the slope toe. The heavy rainfall patterns significantly affect the temporal and spatial evolution of pore water pressure, displacement and safety factors of the slope. Pore water pressure and displacement show a positive correlation with the rainfall intensity at various times during heavy rainfall events. The pre-peak rainfall pattern causes the largest decrease in the safety factor of the slope, and the slope failure occurs earlier, which is the most detrimental to the stability of the slope. The rainfall intensity is inversely proportional to the safety factor. As the rainfall intensity increases, the decrease in the slope’s safety factor becomes more significant, and the time required for slope instability is also shortened. The results of this study provide a scientific basis for analyzing rainfall-induced slope instability and failure. Full article

Background/objectives: Being overweight increases the predisposition to obesity and type 2 diabetes (T2D), which significantly elevate stroke risk and the likelihood of severe post-stroke disability. Dietary nitrate (NO₃⁻) supplementation can mitigate obesity and metabolic impairments, making it a promising approach to halt overweight people from developing overt obesity/T2D, thereby potentially also improving stroke outcome. We determined whether NO3⁻ supplementation prevents overweight mice from progressing into obesity and T2D and whether this intervention improves stroke outcome. Methods: An overweight condition was induced via 6 weeks of a high-fat diet (HFD), after which animals were randomized to either a HFD or a HFD with NO₃⁻ supplementation. After 24 weeks, when HFD-mice without NO₃⁻ developed obesity and T2D, all animals were subjected to transient middle cerebral artery occlusion and stroke outcome was assessed via behavioral testing and infarct size. The effect of NO₃⁻ on post-stroke neuroinflammation, neurogenesis, and neovascularization was analyzed by immunohistochemistry. Results: Sustained NO₃⁻ supplementation in overweight mice did not prevent obesity or insulin resistance. However, it attenuated weight gain, prevented hyperglycemia, and significantly improved functional recovery after stroke, without affecting infarct size. Moreover, NO₃⁻ decreased post-stroke neuroinflammation by reducing microglial infiltration. NO₃⁻ did not affect stroke-induced neurogenesis or vascularization. Conclusion: These results highlight the potential of NO₃⁻ supplementation to prevent metabolic impairment in the overweight population and improve stroke prognosis in this large group of people at risk of stroke and severe stroke sequelae. Full article

The Gliomas account for 81% of all malignant central nervous system tumors and are classified by WHO into four grades of malignancy. Glioblastoma multiforme (GBM), the most common grade IV glioma, exhibits an extremely aggressive phenotype and a dismal five-year survival rate of only 6%, underscoring the urgent need for novel therapeutic approaches. Immunotherapy has emerged as a promising strategy, and photodynamic therapy (PDT) in particular has attracted attention for its dual cytotoxic and immunostimulatory effects. In GBM models, PDT induces immunogenic cell death characterized by the release of damage-associated molecular patterns (DAMPs), which promote antigen presentation and activate T cell responses. Additionally, PDT transiently increases blood–brain barrier permeability, facilitating immune cell infiltration into the tumor microenvironment, and enhances clearance of waste products via stimulation of meningeal lymphatic vessels. Importantly, PDT can reprogram or inactivate immunosuppressive tumor-associated macrophages, thereby counteracting the pro-tumoral microenvironment. Despite these encouraging findings, further preclinical and clinical studies are required to elucidate PDT’s underlying immunological mechanisms fully and to optimize treatment regimens that maximize its efficacy as part of integrated immunotherapeutic strategies against GBM. Full article

Resin Transfer Molding (RTM) is a widely used composite manufacturing process where liquid resin is injected into a closed mold filled with a fibrous preform. By applying this process, large pieces with complex shapes can be produced on an industrial scale, presenting excellent properties and quality. A true physical phenomenon occurring in the RTM process, especially when using vegetable fibers, is related to the absorption of resin by the fiber during the infiltration process. The real effect is related to the slowdown in the advance of the fluid flow front, increasing the mold filling time. This phenomenon is little explored in the literature, especially for non-isothermal conditions. In this sense, this paper does a numerical study of the liquid injection process in a closed and heated mold. The proposed mathematical modeling considers the radial, three-dimensional, and transient flow, variable injection pressure, and fluid viscosity, including the effect of liquid fluid absorption by the reinforcement (fiber). Simulations were carried out using Computational Fluid Dynamic tools. The numerical results of the filling time were compared with experimental results, and a good approximation was obtained. Further, the pressure, temperature, velocity, and volumetric fraction fields, as well as the transient history of the fluid front position and injection fluid volumetric flow rate, are presented and analyzed. Full article

Calcium (Ca²⁺) signaling is a fundamental regulatory mechanism controlling essential processes in the endothelium, vascular smooth muscle cells (VSMCs), and the extracellular matrix (ECM), including maintaining the endothelial barrier, modulation of vascular tone, and vascular remodeling. Cytosolic free Ca²⁺ concentration is tightly regulated by a balance between Ca²⁺ mobilization mechanisms, including Ca²⁺ release from the intracellular stores in the sarcoplasmic/endoplasmic reticulum and Ca²⁺ entry via voltage-dependent, transient-receptor potential, and store-operated Ca²⁺ channels, and Ca²⁺ elimination pathways including Ca²⁺ extrusion by the plasma membrane Ca²⁺-ATPase and Na⁺/Ca²⁺ exchanger and Ca²⁺ re-uptake by the sarco(endo)plasmic reticulum Ca²⁺-ATPase and the mitochondria. Some cell membranes/organelles are multifunctional and have both Ca²⁺ mobilization and Ca²⁺ removal pathways. Also, the individual Ca²⁺ handling pathways could be integrated to function in a regenerative, capacitative, cooperative, bidirectional, or reciprocal feed-forward or feed-back manner. Disruption of these pathways causes dysregulation of the Ca²⁺ signaling dynamics and leads to pathological cardiovascular conditions such as hypertension, coronary artery disease, atherosclerosis, and vascular calcification. In the endothelium, dysregulated Ca²⁺ signaling impairs nitric oxide production, reduces vasodilatory capacity, and increases vascular permeability. In VSMCs, Ca²⁺-dependent phosphorylation of the myosin light chain and Ca²⁺ sensitization by protein kinase-C (PKC) and Rho-kinase (ROCK) increase vascular tone and could lead to increased blood pressure and hypertension. Ca²⁺ activation of matrix metalloproteinases causes collagen/elastin imbalance and promotes vascular remodeling. Ca²⁺-dependent immune cell activation, leukocyte infiltration, and cholesterol accumulation by macrophages promote foam cell formation and atherosclerotic plaque progression. Chronic increases in VSMCs Ca²⁺ promote phenotypic switching to mesenchymal cells and osteogenic transformation and thereby accelerate vascular calcification and plaque instability. Emerging therapeutic strategies targeting these Ca²⁺-dependent mechanisms, including Ca²⁺ channel blockers and PKC and ROCK inhibitors, hold promise for restoring Ca²⁺ homeostasis and mitigating vascular disease progression. Full article

T lymphocytes infiltrate the CNS in response to murine cytomegalovirus (MCMV) infection and form a pool of long-lived brain tissue-resident memory T-cells (bT_RMs), which display markers of residency (i.e., CD103, CD69, CD49a). However, the functional role of these bT_RMs is still unknown. By 30 days postinfection, a latent viral brain infection was established, as indicated by absence of viral transcripts (IE1, E1, and gB) produced during productive infection. Following intracerebroventricular injection of either depleting α-CD8 Ab (clone YTS169.4) or α-CD103-sap (clone IT50) into the brain, 90–95% T-cell depletion was achieved. Using luciferase-expressing mice, we observed recommenced imaging signals indicative of de novo MCMV IE promoter activity in depleted animals. Surprisingly, using an explant assay, we efficiently recovered reactivatable, infectious virus from untreated, latent animals, but not from those depleted of bT_RMs (viral recovery in explants was reduced from 100% to 50% by day 21). We identified Lgals3 (galectin 3), Gpnmb (glycoprotein nonmetastatic melanoma protein B) and Hmox1 (heme oxygenase 1) as genes that were most upregulated in bT_RM-depleted groups. When bT_RMs were depleted, there was transient expression of viral IE genes which resulted in antiviral microglia with a phagocytic, disease-associated (DAM) or neurodegenerative (MGnD) phenotype. These data provide new insights into the role of bT_RMs in controlling both CNS reactivation and driving microglial phenotypes. Full article

This study investigates rainfall-induced slope instability in fine-grained clayey soils through a probabilistic and sensitivity analysis framework that integrates spatial variability. Moving beyond traditional deterministic methods, Monte Carlo simulations were employed to quantify uncertainty in geotechnical parameters—unit weight, cohesion, and friction angle—modeled as random fields with a 1 m spatial resolution. This approach realistically captures natural soil heterogeneity and its influence on slope behavior during rainfall events. Transient seepage and slope stability analyses were performed using SEEP/W and SLOPE/W, respectively, with the Spencer method ensuring full equilibrium. This study examined how slope height, inclination, rainfall intensity and duration, and soil properties affect the factor of safety (FS). The results showed that higher rainfall intensity and longer durations significantly increase failure risk. For example, under 9 mm/h rainfall for 48 h, slopes taller than 10 m at 45° inclination exhibited failure probabilities over 30%. At 20 m, FS dropped to 0.68 with a 100% probability of failure. Sensitivity analysis confirmed cohesion and friction angle as key stabilizing factors, though their impact diminishes with infiltration. A dataset of 9984 slope scenarios was generated, supporting future machine learning applications for risk assessment and climate-resilient slope design. Full article

Cannabis sativa produces pharmacologically valuable cannabinoids. In this study, we developed and optimized a transient transformation system using Cannabis sativa ‘Cheungsam’ to facilitate gene functional analysis. Various experimental conditions, including plant developmental stages, light conditions, Agrobacterium strains, tissue types, and physical treatments such as sonication and vacuum infiltration, were systematically evaluated using GUS histochemical staining and qPCR analysis. Among these, 7-day-old seedlings cultured under dark conditions and transformed with the GV3101 strain exhibited high transformation efficiency. Leaf tissue showed a higher GUS staining proportion and GUS staining area compared to hypocotyl and cotyledon tissues. The application of a combination of sonication and vacuum infiltration techniques resulted in the most intense GUS expression. Using the optimized protocol, we introduced a recombinant vector carrying CsCBDAS2, a key gene in cannabidiol (CBD) biosynthesis. qPCR analysis revealed that CsCBDAS2 overexpression led to significant upregulation of multiple upstream CBD biosynthetic genes (CsOAC, CsGOT, CsPT1, CsPT4, CsCBDAS1, and CsCBDAS2) and the transcription factor (TF) CsWRKY20, suggesting coordinated co-expression and potential involvement of a transcriptional feedback loop. These results demonstrate the effectiveness of our transient transformation system and provide insights into the regulatory mechanisms of cannabinoid biosynthesis in cannabis. Full article

Buckwheat is a promising crop with grains that are rich in nutrients and bioactive compounds. Genome sequence data for common and Tartary buckwheat have recently become available. Currently, there is a critical need for the development of a simple and reliable transient gene expression protocol, as well as a stable genetic transformation method, to facilitate metabolic engineering of bioactive compounds, functional analysis of genes, targeted editing, and, in a long-term perspective, to accelerate the breeding process in buckwheat. In this paper, we report optimized methods for Agrobacterium-mediated transient and stable transformation of Fagopyrum esculentum and F. tartaricum. Leaf and cotyledon tissues were infiltrated with an A. tumefaciens-bearing construct containing eGFP and GUS reporter genes. Histochemical staining and Western blotting were used to confirm the expression of reporter proteins. We also demonstrate the usefulness of the developed method for engineering the gramine biosynthetic pathway in buckwheat. HvAMIS and HvNMT genes were transiently expressed in buckwheat leaves, and the de novo production of gramine was confirmed by LC-MS. Moreover, in planta genetic transformation of common and Tartary buckwheat with a reporter gene (eGFP) and selectable marker gene (NptII) was achieved by Agrobacterium-mediated vacuum infiltration. Genomic integration of the construct was confirmed by polymerase chain reaction (PCR), whereas the production of eGFP was confirmed by fluorescence microscopy. Full article

The reduction in the stability of rock slopes due to rainfall is a significant issue in tropical regions. Unsaturated soil, commonly found on hill slopes, provides higher shear strength compared to saturated soil due to matric suction. Soil moisture plays a crucial role in determining slope stability during rainfall events, yet it is often overlooked in geotechnical engineering projects. This study integrates both steady-state and transient analyses to examine how rainfall intensity affects the stability of a rock slope near a tunnel portal. Transient seepage analysis was conducted using SEEP/W to simulate changes in pore water pressure (PWP) resulting from rainfall infiltration under historical and future precipitation conditions. The analysis considers medium (SSP245) and worst-case (SSP585) climate change scenarios as per Coupled Model Intercomparison Project Phase 6 (CMIP6). The findings underscore the significant impact of rainfall-induced infiltration on slope stability and highlight the importance of incorporating soil moisture dynamics in slope stability assessments. The safety factor, initially 1.54 before accounting for rainfall effects, decreases to 1.34 when the effects of rainfall are included. Full article