A method for predicting needle insertion deflection in soft tissue based on cutting force identification.

The deflection modeling during the insertion of bevel-tipped flexible needles into soft tissues is crucial for robot-assisted flexible needle insertion into specific target locations within the human body during percutaneous biopsy surgery. This paper proposes a mechanical model based on cutting force identification to predict the deflection of flexible needles in soft tissues. Unlike other models, this method does not require measuring Young's modulus (E) and Poisson's ratio (ν) of tissues, which require complex hardware to obtain. In the model, the needle puncture process is discretized into a series of uniform-depth puncture steps. The needle is simplified as a cantilever beam supported by a series of virtual springs, and the influence of tissue stiffness on needle deformation is represented by the spring stiffness coefficient of the virtual spring. By theoretical modeling and experimental parameter identification of cutting force, the spring stiffness coefficients are obtained, thereby modeling the deflection of the needle. To verify the accuracy of the proposed model, the predicted model results were compared with the deflection of the puncture experiment in polyvinyl alcohol (PVA) gel samples, and the average maximum error range predicted by the model was between 0.606 ± 0.167 mm and 1.005 ± 0.174 mm, which showed that the model can successfully predict the deflection of the needle. This work will contribute to the design of automatic control strategies for needles.

MOTAI: A Novel Method for the Study of O-GalNAcylation and Complex O-Glycosylation in Cancer.

The Tn antigen, an immature truncated O-glycosylation, is a promising biomarker for cancer detection and diagnosis. However, reliable methods for analyzing O-GalNAcylation and complex O-glycosylation are lacking. Here, we develop a novel method, MOTAI, for the sequential analysis of O-glycosylation using different O-glycoproteases. MOTAI conjugates glycopeptides on a solid support and releases different types of O-glycosylation through sequential enzymatic digestion by O-glycoproteases, including OpeRATOR and IMPa. Because OpeRATOR has less activity on O-GalNAcylation, MOTAI enriches O-GalNAcylation for subsequent analysis. We demonstrate the effectiveness of MOTAI by analyzing fetuin O-glycosylation and Jurkat cell lines. We then apply MOTAI to analyze colorectal cancer and benign colorectal polyps. We identify 32 Tn/sTn-glycoproteins and 43 T/sT-glycoproteins that are significantly increased in tumor tissues. Gene Ontology analysis reveals that most of these proteins are ECM proteins involved in the adhesion process of the intercellular matrix. Additionally, the protein disulfide isomerase CRELD2 has a significant difference in Tn expression, and the abnormally glycosylated T345 and S349 O-glycosylation sites in cancer group samples may promote the secretion of CRELD2 and ultimately tumorigenesis through ECM reshaping. In summary, MOTAI provides a powerful new tool for the in-depth analysis of O-GalNAcylation and complex O-glycosylation. It also reveals the upregulation of Tn/sTn-glycoproteins in colorectal cancer, which may provide new insights into cancer biology and biomarker discovery.

A back propagation neural network based respiratory motion modelling method.

BACKGROUND: This study presents the development of a backpropagation neural network-based respiratory motion modelling method (BP-RMM) for precisely tracking arbitrary points within lung tissue throughout free respiration, encompassing deep inspiration and expiration phases. METHODS: Internal and external respiratory data from four-dimensional computed tomography (4DCT) are processed using various artificial intelligence algorithms. Data augmentation through polynomial interpolation is employed to enhance dataset robustness. A BP neural network is then constructed to comprehensively track lung tissue movement. RESULTS: The BP-RMM demonstrates promising accuracy. In cases from the public 4DCT dataset, the average target registration error (TRE) between authentic deep respiration phases and those forecasted by BP-RMM for 75 marked points is 1.819 mm. Notably, TRE for normal respiration phases is significantly lower, with a minimum error of 0.511 mm. CONCLUSIONS: The proposed method is validated for its high accuracy and robustness, establishing it as a promising tool for surgical navigation within the lung.

Development and validation of machine learning models and nomograms for predicting the surgical difficulty of laparoscopic resection in rectal cancer.

BACKGROUND: The objective of this study is to develop and validate a machine learning (ML) prediction model for the assessment of laparoscopic total mesorectal excision (LaTME) surgery difficulty, as well as to identify independent risk factors that influence surgical difficulty. Establishing a nomogram aims to assist clinical practitioners in formulating more effective surgical plans before the procedure. METHODS: This study included 186 patients with rectal cancer who underwent LaTME from January 2018 to December 2020. They were divided into a training cohort (n = 131) versus a validation cohort (n = 55). The difficulty of LaTME was defined based on Escal's et al. scoring criteria with modifications. We utilized Lasso regression to screen the preoperative clinical characteristic variables and intraoperative information most relevant to surgical difficulty for the development and validation of four ML models: logistic regression (LR), support vector machine (SVM), random forest (RF), and decision tree (DT). The performance of the model was assessed based on the area under the receiver operating characteristic curve(AUC), sensitivity, specificity, and accuracy. Logistic regression-based column-line plots were created to visualize the predictive model. Consistency statistics (C-statistic) and calibration curves were used to discriminate and calibrate the nomogram, respectively. RESULTS: In the validation cohort, all four ML models demonstrate good performance: SVM AUC = 0.987, RF AUC = 0.953, LR AUC = 0.950, and DT AUC = 0.904. To enhance visual evaluation, a logistic regression-based nomogram has been established. Predictive factors included in the nomogram are body mass index (BMI), distance between the tumor to the dentate line ≤ 10 cm, radiodensity of visceral adipose tissue (VAT), area of subcutaneous adipose tissue (SAT), tumor diameter >3 cm, and comorbid hypertension. CONCLUSION: In this study, four ML models based on intraoperative and preoperative risk factors and a nomogram based on logistic regression may be of help to surgeons in evaluating the surgical difficulty before operation and adopting appropriate responses and surgical protocols.

A spatial registration method based on 2D-3D registration for an augmented reality spinal surgery navigation system.

BACKGROUND: In order to provide accurate and reliable image guidance for augmented reality (AR) spinal surgery navigation, a spatial registration method has been proposed. METHODS: In the AR spinal surgery navigation system, grayscale-based 2D/3D registration technology has been used to register preoperative computed tomography images with intraoperative X-ray images to complete the spatial registration, and then the fusion of virtual image and real spine has been realised. RESULTS: In the image registration experiment, the success rate of spine model registration was 90%. In the spinal model verification experiment, the surface registration error of the spinal model ranged from 0.361 to 0.612 mm, and the total average surface registration error was 0.501 mm. CONCLUSION: The spatial registration method based on 2D/3D registration technology can be used in AR spinal surgery navigation systems and is highly accurate and minimally invasive.

Deciphering Protein O-GalNAcylation: Method Development and Disease Implication.

Mucin-type O-glycosylation is an important protein post-translational modification that is abundantly expressed on cell surface proteins. Protein O-glycosylation plays a variety of roles in cellular biological functions including protein structure and signal transduction to the immune response. Cell surface mucins are highly O-glycosylated and are the main substance of the mucosal barrier that protects the gastrointestinal or respiratory tract from infection by pathogens or microorganisms. Dysregulation of mucin O-glycosylation may impair mucosal protection against pathogens that can invade cells to trigger infection or immune evasion. Truncated O-glycosylation, also known as Tn antigen or O-GalNAcylation, is highly upregulated in diseases such cancer, autoimmune disorders, neurodegenerative diseases, and IgA nephropathy. Characterization of O-GalNAcylation helps decipher the role of Tn antigen in physiopathology and therapy. However, the analysis of O-glycosylation, specifically the Tn antigen, remains challenging due to the lack of reliable enrichment and identification assays compared to N-glycosylation. Here, we summarize recent advances in analytical methods for O-GalNAcylation enrichment and identification and highlight the biological role of the Tn antigen in various diseases and the clinical implications of identifying aberrant O-GalNAcylation.

Isolation of biofluids from tissues using a vacuum-assisted filtration biomedical device.

A biopsy is usually used to remove a piece of tissue from a patient for laboratory testing. The interstitial fluid is taken out at the same time as the tissue sample. Since interstitial fluid flows between cells and capillaries in tissues, similar to blood plasma, it is necessary to separate interstitial fluid from tissues in order to study them separately. Vacuum blood sampling has been used to draw blood into vacuum-sealed tubes, while interstitial fluid can be removed directly from the skin using microneedles with standard pumps. However, no methods are available to separate blood or interstitial fluid from the tissue itself for molecular characterization. In this study, we designed a biomedical device that can separate interstitial fluid from tissue using a vacuum-assisted filtration method. The device has a chamber that collects fluid extracted from the tissue that remains on top of the filter. We characterized the weight change and glycan profiles of tissues before and after vacuum-assisted filtration. The results demonstrate that the biomedical device can remove interstitial fluid and facilitate the analysis of tissue-specific molecules while minimizing information from the interstitial fluid.

Quantitative analysis of fucosylated glycoproteins by immobilized lectin-affinity fluorescent labeling.

Human biofluids are often used to discover disease-specific glycosylation, since abnormal changes in protein glycosylation can discern physiopathological states. Highly glycosylated proteins in biofluids make it possible to identify disease signatures. Glycoproteomic studies on saliva glycoproteins showed that fucosylation was significantly increased during tumorigenesis and that glycoproteins became hyperfucosylated in lung metastases, and tumor stage is associated with fucosylation. Quantification of salivary fucosylation can be achieved by mass spectrometric analysis of fucosylated glycoproteins or fucosylated glycans; however, the use of mass spectrometry is non-trivial for clinical practice. Here, we developed a high-throughput quantitative method, lectin-affinity fluorescent labeling quantification (LAFLQ), to quantify fucosylated glycoproteins without relying on mass spectrometry. Lectins with a specific affinity for fucoses are immobilized on the resin and effectively capture fluorescently labeled fucosylated glycoproteins, which are further quantitatively characterized by fluorescence detection in a 96-well plate. Our results demonstrated that serum IgG can be accurately quantified by lectin and fluorescence detection. Quantification in saliva showed significantly higher fucosylation in lung cancer patients compared to healthy controls or other non-cancer diseases, suggesting that this method has the potential to quantify stage-related fucosylation in lung cancer saliva.

Surgical Navigation System for Hypertensive Intracerebral Hemorrhage Based on Mixed Reality.

Hypertensive intracerebral hemorrhage (HICH) is an intracerebral bleeding disease that affects 2.5 per 10,000 people worldwide each year. An effective way to cure this disease is puncture through the dura with a brain puncture drill and tube; the accuracy of the insertion determines the quality of the surgery. In recent decades, surgical navigation systems have been widely used to improve the accuracy of surgery and minimize risks. Augmented reality- and mixed reality-based surgical navigation is a promising new technology for surgical navigation in the clinic, aiming to improve the safety and accuracy of the operation. In this study, we present a novel multimodel mixed reality navigation system for HICH surgery in which medical images and virtual anatomical structures can be aligned intraoperatively with the actual structures of the patient in a head-mounted device and adjusted when the patient moves in real time while under local anesthesia; this approach can help the surgeon intuitively perform intraoperative navigation. A novel registration method is used to register the holographic space and serves as an intraoperative optical tracker, and a method for calibrating the HICH surgical tools is used to track the tools in real time. The results of phantom experiments revealed a mean registration error of 1.03 mm and an average time consumption of 12.9 min. In clinical usage, the registration error was 1.94 mm, and the time consumption was 14.2 min, showing that this system is sufficiently accurate and effective for clinical application.

Characterization of Pathway-Specific Regulator NigR for High Yield Production of Nigericin in Streptomyces malaysiensis F913.

Nigericin is a polyether antibiotic with potent antibacterial, antifungal, antimalarial and anticancer activity. NigR, the only regulator in the nigericin biosynthetic gene cluster in Streptomyces malaysiensis F913, was identified as a SARP family regulator. Disruption of nigR abolished nigericin biosynthesis, while complementation of nigR restored nigericin production, suggesting that NigR is an essential positive regulator for nigericin biosynthesis. Overexpression of nigR in Streptomyces malaysiensis led to significant increase in nigericin production compared to the wild-type strain. Nigericin production in the overexpression strain was found to reach 0.56 g/L, which may be the highest nigericin titer reported to date. Transcriptional analysis suggested that nigR is required for the transcription of structural genes in the nig gene cluster; quantitative RT-PCR analysis revealed that the expression of structural genes was upregulated in the nigR overexpression strain. Our study suggested that NigR acts in a positive manner to modulate nigericin production by activating transcription of structural genes and provides an effective strategy for scaling up nigericin production.

Personalized virtual reality simulation training system for percutaneous needle insertion and comparison of zSpace and vive.

PURPOSE: In this research, we present a personalized simulation training system for percutaneous needle insertion based on virtual reality (VR). Within this system, surgeons can become more familiar with real surgical procedures, thereby reducing errors that may occur in real surgery. Additionally, different VR technologies, i.e., zSpace and Vive, were compared to provide surgeons with a better surgical training environment. METHODS: and Methods A VR system combined with the treatment planning system was developed to create personalized patient training environment. An evaluation study recruiting twenty novices was performed to demonstrate the system. Each participant performed six independent needle placements using the VR system. Placement time was recorded. Placement error was defined as the distance from the needle tip to the target center. The participants completed a seven-point Likert scale questionnaire after the simulation. RESULTS: Compared with Vive, using zspace reduces the placement time (from 90.32 s to 68.94 s). The placement error using zSpace and Vive was similar (1.27 ± 0.68 mm, 1.56 ± 0.81 mm, respectively). The questionnaire survey results show that most participants are highly satisfied with the training effect of the VR system. Participants prefer the operation mode and convenience of zSpace but think that the immersion of Vive is better. CONCLUSIONS: The personalized virtual reality surgical training system was effective as a training system for percutaneous needle insertion. The system based on zSpace had a shorter placement time while maintaining placement accuracy compared to the system based on Vive. The system based on zSpace achieved higher satisfaction in most aspects except for immersion.

Melatonin enhances the antioxidant capacity to rescue the honey bee Apis mellifera from the ecotoxicological effects caused by environmental imidacloprid.

Imidacloprid severely poisons the nontarget insect honey bee Apis mellifera. Few treatments are available to mitigate the adverse effects of imidacloprid. The primary concern is that the molecular understanding of imidacloprid toxicity is not comprehensive enough. Oxidative stress is the primary pathophysiological mechanism by which pesticides cause high mortality. Our pilot study found for the first time that imidacloprid stimulates bee brains to secrete melatonin, a free radical scavenger. However, the molecular basis for imidacloprid toxicity and the role of melatonin in coping with imidacloprid have not been systematically investigated in bees. This study administered an environmental dose of imidacloprid (36 ng/bee) orally to A. mellifera. The detoxification gene cytochrome P450 CYP4G11 was significantly induced. However, potent cytotoxicity of imidacloprid suppressed the expression of the antioxidants catalase (CAT) and thioredoxin reductase (TrxR), and the activity of guaiacol peroxidase (GPX), superoxide dismutase (SOD), and reduced glutathione (GSH) was not induced. The levels of reactive oxygen species (ROS) and the lipid peroxidation marker malondialdehyde (MDA) were increased. The expression of the apoptotic genes cysteinyl aspartate specific proteinase (Caspase-3) and apoptosis inducing factor (AIF) increased, and the apoptotic features of midgut cells were prominently apparent. These results suggest that imidacloprid disrupts the bee antioxidant system, causing severe oxidative stress and tissue damage and ultimately leading to apoptosis. Significantly, however, imidacloprid exposure also stimulated bee brains to continuously secrete melatonin. Further preadministration of exogenous melatonin (200 ng/bee) orally to bees significantly reversed and enhanced the activity of the imidacloprid-suppressed antioxidants CAT, SOD, and GSH, which allowed imidacloprid-induced ROS accumulation to be effectively alleviated. The MDA content, apoptotic genes Caspase-3 and AIF, and detoxification gene CYPG411 expression were restored to normalization; midgut cell damage, apoptosis, and mortality were significantly reduced. These findings strongly suggest that melatonin enhanced bee antioxidant capacity, thus attenuating oxidative stress and apoptosis to confer imidacloprid tolerance to honey bees. Melatonin secretion may be a defense mechanism to mitigate imidacloprid toxicity.

An integrated navigation system based on a dedicated breast support device for MRI-guided breast biopsy.

PURPOSE: Breast cancer is currently the cancer type with the highest incidence in the world, and it is extremely harmful to women's health. MRI-guided breast biopsy is a common method in clinical examination of breast cancer. However, traditional breast biopsy is less accurate and takes a long time. In this study, an integrated navigation system (INS) based on a dedicated breast support device (DBSD) was proposed to assist doctors in biopsy. METHODS: The grid-shaped DBSD can reduce the displacement and deformation of the breast during the biopsy operation and is convenient for puncture. The robot system based on the DBSD is designed to assist doctors in performing puncture action. The software system has functions such as registration, path planning, and real-time tracking of biopsy needles based on the DBSD, which can assist doctors in completing the entire biopsy procedure. A series of experiments are designed to verify the feasibility and accuracy of the system. RESULTS: Experiments prove that the robot system has reasonable structure and meets the requirements of MR compatibility. The latency of the INS during intraoperative navigation is 0.30 ± 0.03 s. In the phantom puncture experiment, the puncture error under the navigation of the INS is 1.04 ± 0.15 mm. CONCLUSION: The INS proposed in this paper can be applied to assist doctors in breast biopsy in MR environment, improve the accuracy of biopsy and shorten the time of biopsy. The experimental results show that the system is feasible and accurate.

Augmented reality surgical navigation system based on the spatial drift compensation method for glioma resection surgery.

BACKGROUND: The number of patients who suffer from glioma has been increasing, and this malignancy is a serious threat to human health. The mainstream treatment for glioma is surgical resection; therefore, accurate resection can improve postoperative patient recovery. PURPOSE: Many studies have investigated surgical navigation guided by mixed reality, with good outcomes. However, the limitations of mixed reality, such as spatial drift caused by environmental changes, limit its clinical application. Therefore, we present a mixed reality surgical navigation system for glioma resection. Preoperative information can be fused precisely with the real patient with the spatial compensation method to achieve clinically suitable accuracy. METHODS: A head-mounted device was used to display virtual information, and a markerless spatial registration method was applied to precisely align the virtual anatomy with the real patient preoperatively. High-accuracy preoperative and intraoperative movement and spatial drift compensation methods were used to increase the positional accuracy of the mixed reality-guided glioma resection system when the patient's head is fixed to the bed frame. Several experiments were designed to validate the accuracy and efficacy of this system. RESULTS: Phantom experiments were performed to test the efficacy and accuracy of this system under ideal conditions, and clinical tests were conducted to assess the performance of this system in clinical application. The accuracy of spatial registration was 1.18 mm in the phantom experiments and 1.86 mm in the clinical application. CONCLUSIONS: Herein, we present a mixed reality-based multimodality-fused surgical navigation system for assisting surgeons in intuitively identifying the glioma boundary intraoperatively. The experimental results indicate that this system has suitable accuracy and efficacy for clinical usage.

Magnetically Boosted Generation of Intracellular Reactive Oxygen Species toward Magneto-Photodynamic Therapy.

The generation of reactive oxygen species (ROS) in photodynamic therapy (PDT) involves excited-state intermediates with both singlet and triplet spin configurations, which provides possibilities to modulate the ROS production in PDT under an external magnetic field. Here, we present that magnetically modulated ROS production can promote PDT efficacy and develop a magnetic-field-assisted PDT (magneto-PDT) method for effectively and selectively killing cancer cells. The photosensitization reaction between excited-state riboflavin and oxygen molecules is influenced by the applied field, and the overall magnetic field effect (MFE) shows a moderate increase at a low field (<1000 G) and then a boost up to the saturation ∼100% at a high field (>1000 G). It is found that the spin precession occurring in radical ion pairs (electron transfer from riboflavin to oxygen) facilitates the O2•- generation at the low field. In comparison, the spin splitting in an encounter complex (energy transfer from riboflavin to oxygen) benefits the production of 1O2 species at the high field. The field modulation on the two types of ROS in PDT, i.e., O2•- and 1O2, is also demonstrated in living cells. The magneto-PDT strategy shows the capability to inhibit the proliferation of cancer cells (e.g., HeLa, RBL-2H3, and MCF-7) effectively and selectively, which reveals the potential of using the MFE on chemical reactions in biological applications.

DVH-based inverse planning for LDR pancreatic brachytherapy.

PURPOSE: Pancreatic cancer is one of the common types of malignant cancer. Low-dose-rate (LDR) brachytherapy has shown great efficacy in curing pancreatic cancer. However, a long preoperative planning time and an insufficient tumor dose are common issues. In this paper, we present and validate a method for inverse planning using simulated annealing (SA) for the treatment of pancreatic cancer. METHODS: The SA method was used for the inverse planning process. With the help of parallel computation and a quick dose field estimation algorithm. This method allowed inverse planning to be performed quickly. A novel length-control method was used to consider and limit the dose of the organ at risk. The effect of this system was validated by calculating the dose-volume histogram metric and time consumption. RESULTS: Regarding the percentage of the volume receiving 100% of the prescribed dose (V100), this approach yielded an average difference in V100 of 5.01% for the tumor and of 1.32% for the organ at risk in the small tumor group; in the large tumor group, the average difference in V100 was 2.3% for the tumor and - 4.49% for the organ at risk. The average time required for inverse planning was 1.63 ± 0.26 s for small tumors and 3.81 ± 0.51 s for large tumors. Compared with other inverse planning methods, the optimal quality of the plans yielded by this method was further improved. CONCLUSION: This paper presents a new type of inverse planning method for the treatment of pancreatic cancer based on SA. Compared with other LDR inverse planning methods, the method presented here can provide the prescribed dose to the tumor while considering the dose of the organ at risk. Also, the required time is significantly lower than other methods. All the experimental results indicate that this method is ready to be applied in further clinical studies.

An inverse planning simulated annealing algorithm with adaptive weight adjustment for LDR pancreatic brachytherapy.

PURPOSE: The inverse planning simulated annealing (IPSA) algorithm has shown good results in cancer surgical treatment planning. However, an adaptive approach has not well been proposed for different shapes and sizes of tumors. The purpose of this study was to propose an adaptive, efficient and safe algorithm to get high-quality treatment dose planning, which is presented for pancreatic cancer. METHODS: An algorithm employs an optimized IPSA and an adaptive process for adjusting the weight of organs at risk (OAR) and tumor. The algorithm, which was combined with ant colony optimization, was further optimized to reduce the number of needles. It could meet the clinical dose objectives within the tumors, reduce the dose distribution within the OAR and minimize the number of needles. Ten clinical cases were chosen randomly from patients, previously successfully treated in clinic to test our method. The algorithm was validated against clinical cases, using clinically relevant dose parameters. RESULTS: The results were compared with clinical results in ten cases, indicating that the dose distribution within the tumor meets the clinical dose objectives. The dose received by OAR had been greatly reduced, and the number of needles could be reduced by about 50%. It was a significant improvement over the clinical treatment planning. CONCLUSIONS: In this paper, we have devised an algorithm to optimize the treatment planning in brachytherapy. The method in this paper could meet the clinical dose objectives and reduce the difficulty of operation. The results were clinically acceptable. This algorithm is also applicable to other cancers such as lung cancer.

A neuroendoscopic navigation system based on dual-mode augmented reality for minimally invasive surgical treatment of hypertensive intracerebral hemorrhage.

BACKGROUND AND OBJECTIVE: Hypertensive intracerebral hemorrhage is characterized by a high rate of morbidity, mortality, disability and recurrence. Neuroendoscopy has been utilized for treatment as an advanced technology. However, traditional neuroendoscopy allows professionals to see only tissue surfaces, and the field of vision is limited, which cannot provide spatial guidance. In this study, an AR-based neuroendoscopic navigation system is proposed to assist surgeons in locating and clearing hematoma. METHODS: The neuroendoscope can be registered through the vector closed loop algorithm. The single-shot method is designed to register medical images with patients precisely. Real-time AR is realized based on video stream fusion. Dual-mode AR navigation is proposed to provide comprehensive guidance from catheter implantation to hematoma removal. A series of experiments is designed to validate the accuracy and significance of this system. RESULTS: The average root mean square error of the registration between medical images and patients is 0.784 mm, and the variance is 0.1426 mm. The pixel mismatching degrees are less than 1% in different AR modes. In catheter implantation experiments, the average error of distance is 1.28 mm, and the variance is 0.43 mm, while the average error of angles is 1.34°, and the variance is 0.45°. Comparative experiments are also conducted to evaluate the feasibility of this system. CONCLUSION: This system can provide stereo images with depth information fused with patients to guide surgeons to locate targets and remove hematoma. It has been validated to have high accuracy and feasibility.

Imidacloprid activates ROS and causes mortality in honey bees (Apis mellifera) by inducing iron overload.

Imidacloprid, a neonicotinoid pesticide widely used for insect pest control, has become a potential pollutant to pollinators. Previous reports have demonstrated the toxicity of this drug in activating oxidative stress resulting in high mortality in the honey bee Apis mellifera. However, the mechanisms underlying the toxicity of imidacloprid have not been fully elucidated. In this study, sublethal (36 ng/bee) and median lethal (132 ng/bee) doses of imidacloprid were administered to bees. The results showed dose-dependent increases in reactive oxygen species (ROS), Fe2+, and mortality in bees. Notably, imidacloprid also induced upregulation of the gene encoding ferritin (AmFth), which plays a pivotal role in reducing Fe2+ overload. Upregulation of AmFth has been suggested to be closely related to ROS accumulation and high mortality in bees. To confirm the role played by AmFth in imidacloprid-activated ROS, dsAmFth double-strand was orally administered to bees after exposure to imidacloprid. The results revealed aggravated Fe2+ overload, higher ROS activation, and elevated mortality in the bees, indicating that imidacloprid activated ROS and caused mortality in the bees, probably by inducing iron overload. This study helps to elucidate the molecular mechanisms underlying the toxicity of imidacloprid from the perspective of iron metabolism.

Characterizing the Xenoma of Vairimorpha necatrix Provides Insights Into the Most Efficient Mode of Microsporidian Proliferation.

Microsporidia are a group of obligated intracellular parasites that can infect nearly all vertebrates and invertebrates, including humans and economic animals. Microsporidian Vairimorpha necatrix is a natural pathogen of multiple insects and can massively proliferate by making tumor-like xenoma in host tissue. However, little is known about the subcellular structures of this xenoma and the proliferation features of the pathogens inside. Here, we characterized the V. necatrix xenoma produced in muscle cells of silkworm midgut. In result, the whitish xenoma was initially observed on the 12th day post infection on the outer surface of the midgut and later became larger and numerous. The observation by scanning electronic microscopy showed that the xenoma is mostly elliptical and spindle with dense pathogen-containing protrusions and spores on the surface, which were likely shedding off the xenoma through exocytosis and could be an infection source of other tissues. Demonstrated with transmission electron microscopy and fluorescent staining, the xenoma was enveloped by a monolayer membrane, and full of vesicle structures, mitochondria, and endoplasmic reticulum around parasites in development, suggesting that high level of energy and nutrients were produced to support the massive proliferation of the parasites. Multiple hypertrophic nuclei were found in one single xenoma, indicating that the cyst was probably formed by fusion of multiple muscle cells. Observed by fluorescence in situ hybridization, pathogens in the xenoma were in merongony, sporogony, and octosporogony, and mature stages. And mature spores were pushed to the center while vegetative pathogens were in the surface layer of the xenoma. The V. necatrix meront usually contained two to three nuclei, and sporont contained two nuclei and was wrapped by a thick membrane with high electron density. The V. necatrix sporogony produces two types of spores, the ordinary dikaryotic spore and unicellular octospores, the latter of which were smaller in size and packed in a sporophorous vesicle. In summary, V. necatrix xenoma is a specialized cyst likely formed by fusion of multiple muscle cells and provides high concentration of energy and nutrients with increased number of mitochondria and endoplasmic reticulum for the massive proliferation of pathogens inside.