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In this study, we report the complexities and challenges associated with achieving robust RNA interference (RNAi)-mediated gene knockdown in the mosquitoes Aedes aegypti and Aedes albopictus, a pivotal approach for genetic analysis and vector control. Despite RNAi's potential for species-specific gene targeting, our independent efforts to establish oral delivery of RNAi for identifying genes critical for mosquito development and fitness encountered significant challenges, failing to reproduce previously reported potent RNAi effects. We independently evaluated a range of RNAi-inducing molecules (siRNAs, shRNAs, and dsRNAs) and administration methods (oral delivery, immersion, and microinjection) in three different laboratories. We also tested various mosquito strains and utilized microorganisms for RNA delivery. Our results reveal a pronounced inconsistency in RNAi efficacy, characterized by minimal effects on larval survival and gene expression levels in most instances despite strong published effects for the tested targets. One or multiple factors, including RNase activity in the gut, the cellular internalization and processing of RNA molecules, and the systemic dissemination of the RNAi signal, could be involved in this variability, all of which are barely understood in mosquitoes. The challenges identified in this study highlight the necessity for additional research into the underlying mechanisms of mosquito RNAi to develop more robust RNAi-based methodologies. Our findings emphasize the intricacies of RNAi application in mosquitoes, which present a substantial barrier to its utilization in genetic control strategies. [ABSTRACT FROM AUTHOR]

Hepatocellular carcinoma is the most common primary liver cancer and the fifth most frequently diagnosed cancer worldwide. Most patients with advanced disease are offered non-surgical palliative treatment options. This work explores the first alpha-particle emitting radioembolization for the treatment and monitoring of hepatic tumors. Furthermore, this works demonstrates the first in vivo simultaneous multiple-radionuclide SPECT-images of the complex decay chain of an [225Ac]Ac-labeled agent using a clinical SPECT system to monitor the temporal distribution. A DOTA chelator was modified with a lipophilic moiety and radiolabeled with the α-particle emitter Actinium-225. The resulting agent, [225Ac]Ac-DOTA-TDA, was emulsified in ethiodized oil and evaluated in vivo in mouse model and the VX2 rabbit technical model of liver cancer. SPECT imaging was performed to monitor distribution of the TAT agent and the free daughters. The [225Ac]Ac-DOTA-TDA emulsion was shown to retain within the HEP2G tumors and VX2 tumor, with minimal uptake within normal tissue. In the mouse model, significant improvements in overall survival were observed. SPECT-imaging was able to distinguish between the Actinium-225 agent (Francium-221) and the loss of the longer lived daughter, Bismuth-213. An α-particle emitting TARE agent is capable of targeting liver tumors with minimal accumulation in normal tissue, providing a potential therapeutic agent for the treatment of hepatocellular carcinoma as well as a variety of hepatic tumors. In addition, SPECT-imaging presented here supports the further development of imaging methodology and protocols that can be incorporated into the clinic to monitor Actinium-225-labeled agents. [ABSTRACT FROM AUTHOR]

Alternating magnetic field (AMF) tissue interaction models are generally not validated. Our aim was to develop and validate a coupled electromagnetic and thermal model for estimating temperatures in large organs during magnetic nanoparticle hyperthermia (MNH). Coupled finite element electromagnetic and thermal model validation was performed by comparing the results to experimental data obtained from temperatures measured in homogeneous agar gel phantoms exposed to an AMF at fixed frequency (155 ± 10 kHz). The validated model was applied to a three-dimensional (3D) rabbit liver built from computed tomography (CT) images to investigate the contribution of nanoparticle heating and nonspecific eddy current heating as a function of AMF amplitude. Computed temperatures from the model were in excellent agreement with temperatures calculated using the analytical method (error < 1%) and temperatures measured in phantoms (maximum absolute error <2% at each probe location). The 3D rabbit liver model for a fixed concentration of 5 mg Fe/cm3 of tumor revealed a maximum temperature ∼44 °C in tumor and ∼40 °C in liver at AMF amplitude of ∼12 kA/m (peak). A validated coupled electromagnetic and thermal model was developed to estimate temperatures due to eddy current heating in homogeneous tissue phantoms. The validated model was successfully used to analyze temperature distribution in complex rabbit liver tumor geometry during MNH. In future, model validation should be extended to heterogeneous tissue phantoms, and include heat sink effects from major blood vessels. [ABSTRACT FROM AUTHOR]

Purpose: Many interventional procedures aim at changing soft tissue perfusion or blood flow. One problem at present is that soft tissue perfusion and its changes cannot be assessed in an interventional suite because cone‐beam computed tomography is too slow (it takes 4–10 s per volume scan). In order to address the problem, we propose a novel method called IPEN for Intra‐operative four‐dimensional soft tissue PErfusion using a standard x‐ray system with No gantry rotation. Methods: IPEN uses two input datasets: (a) the contours and locations of three‐dimensional regions‐of‐interest (ROIs) such as arteries and sub‐sections of cancerous lesions, and (b) a series of x‐ray projection data obtained from an intra‐arterial contrast injection to contrast enhancement to wash‐out. IPEN then estimates a time‐enhancement curve (TEC) for each ROI directly from projections without reconstructing cross‐sectional images by maximizing the agreement between synthesized and measured projections with a temporal roughness penalty. When path lengths through ROIs are known for each x‐ray beam, the ROI‐specific enhancement can be accurately estimated from projections. Computer simulations are performed to assess the performance of the IPEN algorithm. Intra‐arterial contrast‐enhanced liver scans over 25 s were simulated using XCAT phantom version 2.0 with heterogeneous tissue textures and cancerous lesions. The following four sub‐studies were performed: (a) The accuracy of the estimated TECs with overlapped lesions was evaluated at various noise (dose) levels with either homogeneous or heterogeneous lesion enhancement patterns; (b) the accuracy of IPEN with inaccurate ROI contours was assessed; (c) we investigated how overlapping ROIs and noise in projections affected the accuracy of the IPEN algorithm; and (d) the accuracy of the perfusion indices was assessed. Results: The TECs estimated by IPEN were sufficiently accurate at a reference dose level with the root‐mean‐square deviation (RMSD) of 0.0027 ± 0.0001 cm−1 or 13 ± 1 Hounsfield unit (mean ± standard deviation) for the homogeneous lesion enhancement and 0.0032 ± 0.0005 cm−1 for the heterogeneous enhancement (N = 20 each). The accuracy was degraded with decreasing doses: The RMSD with homogeneous enhancement was 0.0220 ± 0.0003 cm−1 for 20% of the reference dose level. Performing 3 × 3 pixel averaging on projection data improved the RMSDs to 0.0051 ± 0.0002 cm−1 for 20% dose. When the ROI contours were inaccurate, smaller ROI contours resulted in positive biases in TECs, whereas larger ROI contours produced negative biases. The bias remained small, within ± 0.0070 cm−1, when the Sorenson–Dice coefficients (SDCs) were larger than 0.81. The RMSD of the TEC estimation was strongly associated with the condition of the problem, which can be empirically quantified using the condition number of a matrix Az that maps a vector of ROI enhancement values z to projection data and a weighted variance of projection data: a linear correlation coefficient (R) was 0.794 (P < 0.001). The perfusion index values computed from the estimated TECs agreed well with the true values (R ≥ 0.985, P < 0.0001). Conclusion: The IPEN algorithm can estimate ROI‐specific TECs with high accuracy especially when 3 × 3 pixel averaging is applied, even when lesion enhancement is heterogeneous, or ROI contours are inaccurate but the SDC is at least 0.81. [ABSTRACT FROM AUTHOR]

An increasing number of studies reveal the importance of long noncoding RNAs (lncRNAs) in gene expression control underlying many physiological and pathological processes. However, their role in skin wound healing remains poorly understood. Our study focused on a skin-specific lncRNA, LOC105372576, whose expression was increased during physiological wound healing. In human nonhealing wounds, however, its level was significantly lower compared with normal wounds under reepithelialization. We characterized LOC105372576 as a nuclear-localized, RNAPII-transcribed, and polyade- nylated lncRNA. In keratinocytes, its expression was induced by TGF-β signaling. Knockdown of LOC105372576 and activation of its endogenous transcription, respectively, reduced and increased the motility of keratinocytes and reepithelialization of human ex vivo skin wounds. Therefore, LOC105372576 was termed "wound and keratinocyte migration-associated lncRNA 1" (WAKMAR1). Further study revealed that WAKMAR1 regulated a network of protein-coding genes important for cell migration, most of which were under the control of transcription factor E2F1. Mechanistically, WAKMAR1 enhanced E2F1 expression by interfering with E2F1 promoter methylation through the sequestration of DNA methyltransferases. Collectively, we have identified a lncRNA important for keratinocyte migration, whose deficiency may be involved in the pathogenesis of chronic wounds. [ABSTRACT FROM AUTHOR]

Purpose/objective:The aim of this study was to develop and investigate the properties of a magnetic iron oxide nanoparticle–ethiodised oil formulation for image-guided thermal therapy of liver cancer. Materials and methods:The formulation comprises bionised nano-ferrite (BNF) nanoparticles suspended in ethiodised oil, emulsified with polysorbate 20 (BNF-lip). Nanoparticle size was measured via photon correlation spectroscopy and transmission electron microscopy.In vivothermal therapy capability was tested in two groups of male Foxn1numice bearing subcutaneous HepG2 xenograft tumours. Group I (n = 12) was used to screen conditions for group II (n = 48). In group II, mice received one of BNF-lip (n = 18), BNF alone (n = 16), or PBS (n = 14), followed by alternating magnetic field (AMF) hyperthermia, with either varied duration (15 or 20 min) or amplitude (0, 16, 20, or 24 kA/m). Image-guided fluoroscopic intra-arterial injection of BNF-lip was tested in New Zealand white rabbits (n = 10), bearing liver VX2 tumours. The animals were subsequently imaged with CT and 3 T MRI, up to 7 days post-injection. The tumours were histopathologically evaluated for distribution of BNF-lip. Results:The BNF showed larger aggregate diameters when suspended in BNF-lip, compared to clear solution. The BNF-lip formulation produced maximum tumour temperatures with AMF >20 kA/m and showed positive X-ray visibility and substantial shortening of T1 and T2 relaxation time, with sustained intratumoural retention up to 7 days post-injection. On pathology, intratumoural BNF-lip distribution correlated well with CT imaging of intratumoural BNF-lip distribution. Conclusion:The BNF-lip formulation has favourable thermal and dual imaging capabilities for image-guided thermal therapy of liver cancer, suggesting further exploration for clinical applications. [ABSTRACT FROM PUBLISHER]