Search Results (430)

The mealworm beetle, Tenebrio molitor, is a coleopteran of importance for both immunological and biotechnological research, and it has even been considered as a potential nutraceutical. In recent years, the study of T. molitor has undergone significant development, including immune response, host–parasite interactions, and physiological approaches. However, to perform this type of study, one of the main obstacles is obtaining sufficient hemolymph and viable hemocytes. Thus, we developed a protocol for adult specimens that enables the collection of up to 300 μL of the hemolymph–anticoagulant buffer mixture per specimen, containing approximately 1.5 × 10⁵ hemocytes, with viability ranging from 85% to 90%. The technique involves a double mesothoracic puncture (DMP) and the use of a modified anticoagulant buffer that prevents hemolymph clotting, enabling continuous extravasation and ensuring high yields. Additionally, the hemocytes recovered with this protocol are intact and can be used for subsequent analysis. The hemolymph obtained using this protocol and its applications will help to better understand the processes involving hemolymph and its components in T. molitor, paving the way for further applications. Full article

Background: Subarachnoid hemorrhage (SAH) results from extravasation of blood into the subarachnoid space and is associated with high morbidity and mortality. This study aimed to compare systolic blood pressure variability (SBPV) and intracranial pressure variability (ICPV) in three 8 h intervals during the first 24 h after hospital admission and investigate their associations with discharge disposition and in-hospital mortality. Methods: We retrospectively reviewed charts of adult patients with spontaneous, non-traumatic SAH admitted for at least 24 h from 2016 to 2020. Hourly measurements were recorded for both systolic blood pressure (SBP) and intracranial pressure (ICP), and SBPV and ICPV were measured using successive variation (SV) and standard deviation (SD). Results: A total of 240 patients were included (mean age 57 ± 14.2 years, 64.6% female); 40 (16.7%) died. Univariate analyses showed higher SBP-SV (22.7 ± 13.8) in the first 8 h to be significantly associated with mortality (p = 0.028) and not being discharged home (p = 0.022), compared to those who survived (17.6 ± 7.5) or were discharged home (16.7 ± 5.5). Multivariate logistic regression did not show an association between SBPV and outcomes of interest. Conclusions: Greater early SBPV was associated with mortality in univariate analysis but was not independently predictive after adjustment for clinical severity, suggesting it reflects underlying physiologic instability rather than an independent prognostic factor in SAH. Full article

Background: Coronary artery perforation is a potentially life-threatening complication in 0.2–0.6% of all patients undergoing percutaneous coronary intervention. Despite the ongoing development of technical skills and coronary devices, severe recalcitrant calcified coronary lesions remain a challenge for interventional cardiologists and can carry a potential risk for life-threatening complications, including coronary perforation. Discussion and Conclusion: The algorithm for cardiac vessel perforation could be more comprehensive and cover preventive and predictive measures. It is necessary to take into consideration prompt recognition, implement actions to restabilize the hemodynamic status, understand the source and mechanism of bleeding, and classify the cause of bleeding into proximal, distal, coronary artery bypass graft and collateral vessel, pericardial, myocardial extravasation, and vessel-chamber perforation, as each causality would necessitate a different management strategy for a successful outcome. Imaging information about cardiac vessel injury is useful for a better understanding of the spatial orientation of the coronary vessels. It also helps to detect a hematoma that deteriorates the hemodynamic status without effusion “dry tamponade” and could have a particular role in cardiac interventions to predict and prevent this complication. Full article

Human mesenchymal stem/stromal cells (MSCs) have attracted significant interest in regenerative medicine due to their self-renewal capacity, immunomodulatory functions, multipotency, and relative ease of isolation and expansion. However, several limitations restrict their clinical application, including cellular heterogeneity, challenges in large-scale expansion, and poor in vivo persistence after transplantation. Systemically administered MSCs are rapidly cleared because of limited adhesion, short survival time, and inefficient extravasation, resulting in suboptimal therapeutic efficacy. To overcome these challenges, various strategies have been developed, such as hypoxic preconditioning, biomaterial-based approaches, and genetic modification. Among these, genetic modification represents a particularly powerful and versatile strategy, as it enables targeted enhancement of specific functional properties of MSCs and even the introduction of novel therapeutic capabilities. In this review, we summarize recent advances in genetically engineered MSCs and categorize these approaches into four functional domains: migration, adhesion, secretion, and survival. We further discuss their therapeutic outcomes across diverse disease models in vivo. Collectively, genetic modification substantially enhances the intrinsic therapeutic potential of MSCs and represents a promising direction for the development of next-generation cell-based therapies. Full article

Background/Objectives: Ureteral complications following oblique lumbar interbody fusion (OLIF) are uncommon and are typically attributed to direct mechanical injury. Functional ureteral obstruction without overt ureteral damage remains poorly characterized. We report two cases that provide clinical and intraoperative evidence of a previously underrecognized mechanism of ureteral obstruction associated with anterior cage positioning during OLIF. Case Presentation: Among 180 OLIF procedures performed by a single surgeon, two cases (1.1%) of postoperative or intraoperative ureteral compromise without direct structural injury were identified. In the first case, postoperative imaging revealed hydronephrosis and focal angulation of the left proximal ureter at the level of the interbody cage, without contrast extravasation. The obstruction was managed with double-J ureteral stenting, and serial renal function monitoring confirmed preserved renal function throughout the clinical course. In the second case, retroperitoneal tissue including the ureter was directly observed intraoperatively to be interposed between the anterior longitudinal ligament and the interbody cage during anterior cage placement. Release of the interposed tissue resulted in immediate ureteral decompression without structural damage. Correlation of the postoperative findings in the first case with the intraoperative observations of the second case supports a unified mechanistic explanation: anterior cage advancement may draw retroperitoneal tissue into the cage–anterior longitudinal ligament interface, subjecting the ureter to focal compression or angulation. Conclusions: Functional ureteral obstruction during OLIF may occur secondary to retroperitoneal tissue interposition rather than direct ureteral trauma. Awareness of this mechanism and meticulous protection of the anterior retroperitoneal layer during cage advancement may help prevent avoidable ureteral complications. Full article

Metastasis is a complex, multistep process in which cancer spreads from its original tumor to other sites in the body. During metastasis, tumor cells move away from the primary tumor and intravasate into the lymphatics or circulation. Surviving tumor cells can then extravasate into and remain in distant tissues until they once again begin to proliferate, forming secondary tumors. An excess of reactive oxygen species (ROS) can promote metastasis, dependent on the ROS molecule, its level of excess, and the examined step within the metastatic cascade. Here, we highlight recent studies where ROS promote epithelial-to-mesenchymal transition, cell migration and invasion, circulating tumor cell survival and disseminated tumor cell dormancy. Additionally discussed are novel in vivo ROS detection methods, FDA-approved therapies and clinical trials that manipulate ROS to improve cancer patient survival. Since metastasis is the major cause of cancer-related death, a better understanding of this process and ROS as a contributing factor will help to identify novel targets for inhibition or prevention. Full article

Background/Objectives: Coronary perforations are infrequent but potentially fatal complications during percutaneous coronary intervention (PCI). Interventional management aims to stop extravasation and restore distal flow to prevent tamponade and cardiogenic shock. In current practice, the ping-pong technique is recommended to ensure sealing of the perforation during covered stent delivery. However, this method is complex, time-consuming, and requires a second vascular access. Therefore, we developed a technique that seals the perforation and enables covered stent implantation using a single guide catheter. Methods: This technical note describes a novel technique in which a guide extension catheter (GEC) can be advanced across a vascular perforation after balloon inflation. The insertion of the GEC is made possible by detachment of the balloon hypotube. To minimize leakage, a regular coronary wire introducer needle is attached to the snapped hypotube after GEC loading and continuously inflated to hold nominal pressure. Advancement of the GEC across the perforation immediately limits hemorrhage and facilitates covered stent deployment via a single vascular access. The technique was first evaluated in bench testing and subsequently applied in three illustrative clinical cases at a tertiary referral center using standard, commercially available devices. Results: Bench testing confirmed the reproducibility of the ripping maneuver and successful ballon inflation over enough time to advance the GEC with the introducer married with the ripped hypotube. In all clinical cases, the GEC was successfully advanced across the perforation, allowing prompt covered stent deployment where necessary using a single guide catheter and access site without technical failure. Conclusions: The RIP (Rip and Inflate in Perforations)—technique is a feasible and reproducible alternative to the ping-pong technique. Bench validation and initial clinical application suggest that it may simplify the management of large-vessel perforations while reducing procedural complexity and the need for additional vascular access. Full article

This study investigated the effects of chronic heat stress (HS) on brain injury in broilers and the associated molecular changes. A chronic HS model was established by exposing broilers to 35 °C from 08:00 to 20:00 daily from 21 to 42 days of age, and samples were collected at 28, 35, and 42 days of age. Chronic HS significantly impaired growth performance and was associated with histopathological and ultrastructural alterations in brain tissue. Serum antioxidant enzyme activities and the total antioxidant capacity were significantly reduced, whereas malondialdehyde levels were significantly increased, indicating sustained oxidative stress (OS). Blood–brain barrier (BBB) permeability, assessed by Evans blue extravasation, was significantly higher in HS birds and was accompanied by reduced mRNA expression of the tight junction-related genes ZO-1 and Claudin-5. In addition, chronic HS was associated with increased mRNA expression in genes related to cellular stress, oxidative stress, and inflammation, including key components of the TLR4/MyD88/NF-κB/NLRP3 pathway, as well as decreased expression of IL-4. These findings suggest that chronic HS is associated with enhanced OS, altered neuroinflammatory gene expression, and BBB impairment in the broiler brain. Overall, this study provides evidence that chronic HS is associated with brain injury in broilers and highlights a potential link among OS, inflammation-related transcriptional changes, and BBB dysfunction, thereby offering a basis for further mechanistic and interventional studies. Full article

Brain metastases remain a major problem for cancer patients, impacting their treatment and survival. The pathogenesis of brain metastases is largely unknown. Recent reports indicate that the adhesion molecule protocadherin γ C3 (PCDHGC3) is differentially expressed in various cancer cells and endothelial cells of the blood–brain barrier (BBB), suggesting its involvement in the development of brain metastases. Therefore, we generated a PCDHGC3 knockout (KO) in the triple-negative breast cancer cell line HCC1806 and the malignant melanoma cell line A2058. Control and KO cells were compared using cell proliferation, adhesion and invasion assays, gene expression analyses and matrix metalloproteinase (MMP) activity assays. While the PCDHGC3 KO mutation led to increased proliferation in HCC1806 cells, with no difference observed in A2058, it significantly increased adhesion to in vitro BBB models as well as invasion in both HCC1806 and A2058 KO cell lines. Although changes in mRNA expression of genes involved in metastasis, angiogenesis and cell adhesion were found in PCDHGC3 KO breast cancer and melanoma cells, the number of genes with significantly increased mRNA expression was higher in A2058 KO cells than in HCC1806 KO cells. While the mRNA expression of MMP1 and 2 was increased in A2058 KO cells, no significant changes were found in HCC1806 KO cells. However, increased MMP activity in the cell culture medium was detected in HCC1806 KO cells, while A2058 KO cells showed lower MMP-activity compared to control. These findings provide insights into the role of PCDHGC3 in cancer cell extravasation during metastatic process and identify potential therapeutic targets for further investigation. Full article

Despite the identification of several mediators of arteriogenesis, the growth of natural bypass, the role of lymphocytes, particularly T cells, in this process remains poorly defined. Among these, γδ T cells, which express alternative T cell receptors, have emerged as a key immune component. This study examined the roles of αβ and γδ T cells in arteriogenesis using a murine hindlimb model. While the absence of αβ T cells did not affect arteriogenesis, γδ T cell depletion markedly reduced vascular cell proliferation and perfusion recovery. Early phase analyses revealed impaired mast cell activation, whereas platelet–neutrophil aggregates and neutrophil extravasation were unaffected. In the later proliferative phase, γδ T cell depletion hindered perivascular M2-like (MRC1⁺) macrophage accumulation. Flow cytometric analysis of whole blood in wildtype mice revealed a temporal shift in γδ T cell populations from a CD27⁺/CD39⁻ phenotype, commonly associated with pro-inflammatory functions and IFNγ production, to CD39⁺ phenotypes, which have been linked to anti-inflammatory properties and IL-10 production. In rescue experiments, administration of IFNγ to γδ T cell-depleted mice restored mast cell activation, whereas IL-10 treatment reestablished M2-like (MRC1⁺) macrophage accumulation. These findings collectively identify γδ T cells as critical regulators of both early and late phases of arteriogenesis through coordinated inflammatory and regenerative mechanisms. Full article

Background: Breast cancer brain metastases (BCBM) lack effective treatments, contributing to breast cancer-related morbidity and mortality. Integrating translational animal models and advanced non-invasive imaging can accelerate the development of urgently needed therapies. Method: In this study, we developed an intracarotid method mimicking BCBM and compared it to the stereotactic model in terms of animal welfare, tumour establishment, and blood–brain barrier (BBB) permeability. BCBM was established through intracarotid or stereotactic inoculation of BT474 and MDA-MB-231.Luc2 cells in NMRI nude mice. We utilised magnetic resonance imaging (MRI) and bioluminescence imaging (BLI) to monitor tumour growth and BBB permeability, supported by fluorescent immunohistochemistry for validation. Finally, light sheet microscopy (LSM) was employed to visualise tumour establishment in intact brains. Results: Both inoculation methods achieved a survival rate > 70%, with animals recovering within a week post-surgery. MRI and BLI effectively visualised tumour growth with stereotactic implantation, resulting in single tumours, while intracarotid inoculation led to micro-seeding of up to seven tumours in one brain. Tumour growth was rapid and homogenous in the stereotactic model, whereas the intracarotid model exhibited slower, heterogenous growth. Notably, BBB permeability was significantly higher in small tumours in the stereotactic model when compared to the intracarotid model (p = 0.003). Ex vivo analyses validated these findings with the identification of multiple metastasis in the intracarotid model and single tumours in the stereotactic model. Conclusions: We developed an animal model that closely mimics BCBM, highlighting extravasation and micro-seeding while maintaining animal welfare. Our established imaging protocols enable longitudinal evaluations of BBB permeability and treatment response, creating a translational platform for testing novel anti-cancer therapies. Full article

Background/Objectives: Hyperreflective foci (HRF) are supportive optical coherence tomography (OCT) imaging biomarkers that have been examined for their association with disease progression and severity in various retinal disorders. The accurate identification and segmentation of these tiny structures of lipid extravasation remain complicated because of their small size, class imbalance, similarity in the reflectivity patterns with the surrounding structures and imaging artifacts. While U-Net-based models have promised exceptional results for medical image segmentation, optimal architectural settings and suitable preprocessing methods for HRF detection remain unclear. Methods: This research assessed optimal settings for U-Net-based models for HRF segmentation by evaluating standard U-Net and attention U-Net under different preprocessing regimes. Attention U-Net employed Z-score normalization and contrast-limited adaptive histogram equalization (CLAHE) enhancement with soft dice loss. The standard U-Net was trained on OCT images with CLAHE using focal Tversky loss. A total of 435 fovea-centered OCT B scans with the corresponding, consensus-annotated HRF masks were utilized for this research. Results: The standard U-Net outperformed attention U-Net with a dice score of 0.5207, an AUC of 0.8411, and a recall of 0.6439 on raw OCT images. The attention U-Net with preprocessing (dice: 0.5033, AUC: 0.6987, recall: 0.5391) demonstrated satisfactory performance. The results showed that the U-Net model with CLAHE and focal Tversky loss improved recall by 19.4% relative to the attention U-Net, and this corresponds roughly to a 23% relative decline in false negatives. This indicates increased sensitivity in identifying HRF regions. Conclusions: The best-performing configuration using U-Net-based architectures for segmentation of HRFs combines the standard U-Net model with CLAHE and focal Tversky loss for handling class imbalance. This approach yields relatively higher sensitivity, indicating that the standard U-Net model delivers a simple and robust framework for automated HRF segmentation on the evaluated dataset, promising further validation in broader clinical datasets. Full article

Metastasis is the leading cause of cancer-related death, underscoring the need to elucidate the key mechanisms behind this process. Neutrophil extracellular traps (NETs) have emerged as critical regulators of tumor progression and metastasis. This review summarizes the primary stimuli and signaling pathways that govern NET formation and outlines the mechanistic roles of NET components in tumor growth and metastatic spread. We focus on environmental and tumor microenvironment-derived factors, including psychological stress, tumor-secreted cytokines, and treatment-related responses, that drive NET formation. The involvement of NETs in multiple stages of the metastatic cascade is discussed, including angiogenesis, tumor cell intravasation and extravasation, circulating tumor cell survival, metastatic colonization, and the reactivation of dormant tumor cells. Additionally, we examine how NETs contribute to the establishment of an immunosuppressive microenvironment. Finally, emerging therapeutic strategies targeting NETs are briefly reviewed, highlighting their potential relevance in metastatic cancer treatment. Full article

Injection-related errors remain a common clinical issue and can cause patient discomfort, hematoma formation, and procedural inefficiencies. The visualization of subcutaneous veins using near-infrared (NIR) imaging has gained attention as an effective approach to reducing such errors, as blood exhibits a higher absorption of NIR light than surrounding tissue. In this study, a low-cost, non-invasive vein visualization system is presented to support safer and more accurate venous access. The proposed system integrates an NIR illumination source and a modified webcam within a compact equipment enclosure, allowing subjects to be conveniently examined by placing their arm inside the device. Vein images are automatically acquired using a laptop-based platform, followed by digital image processing techniques for vein enhancement and visualization. Laboratory-scale experiments were conducted on healthy volunteers to evaluate system performance under multiple conditions, including different vein locations (upper and lower arm regions), varying distances between the NIR light source and the arm (15 cm and 20 cm), and ambient illumination interference (light sources on and off). The experimental results demonstrate the successful implementation and reliable operation of the proposed system. Effective vein visualization was achieved across all test conditions, as confirmed by qualitative visual assessment and quantitative image quality metrics, including the Peak Signal-to-Noise Ratio (PSNR) and Mean Squared Error (MSE). Overall, the proposed system offers a practical, accessible, and cost-effective solution for vein visualization, showing strong potential for clinical and experimental applications aimed at reducing injection errors and improving venous access reliability. Full article

Objectives: While several physiological functions of milt peptides have been discovered, the structural characteristics of Theragra chalcogramma milt peptides (TMP) and their anti-fatigue mechanisms remain unclear. Methods: TMP was obtained by hydrolysis via flavor enzyme and alkaline protease, and its structural characteristics were analyzed. A mice model of exercise-induced fatigue was established. The anti-fatigue effect of TMP was evaluated by determining the main biochemical indices in the serum, liver, and skeletal muscle of mice. Additionally, qPCR analysis was conducted to investigate its regulatory effects on relevant energy metabolism pathways. Results: TMP contained 18.2% branched-chain amino acids, with those with molecular weights below 1000 Da accounting for 91.6%. A total of 154 characteristic peptides, such as VPFPR and LPPGR, were identified from TMP, among which 64% of the peptides contained glutamic acid, arginine, or aspartic acid. Molecular docking of potential bioactive peptides to AMP-activated protein kinase (AMPK) revealed binding energies from −9.1 to −5.5 kcal/mol. The exhaustive swimming test showed that oral administration of TMP prolonged the swimming duration. In the fatigue murine model, TMP reduced blood urea nitrogen and blood lactic acid levels while enhancing the content of muscle glycogen. Meanwhile, TMP significantly increased the activity of glutathione peroxidase and superoxide dismutase and reduced the accumulation of malondialdehyde, demonstrating antioxidant properties. Additionally, TMP significantly decreased creatine kinase and lactate dehydrogenase extravasation, thereby protecting muscle tissue, as corroborated by immunohistochemical analyses. Mechanistically, TMP upregulated AMPK and peroxisome proliferator-activated receptor gamma coactivator-1α (PGC-1α) expression, promoting mitochondrial biogenesis via the AMPK/PGC-1α pathway. Conclusions: These findings suggest TMP has potential as a dietary supplement for alleviating physical fatigue. Full article