Novel bispecific nanobody mitigates experimental intestinal inflammation in mice by targeting TNF-α and IL-23p19 bioactivities.

BACKGROUND: Inflammatory bowel diseases (IBDs) pose significant challenges in terms of treatment non-response, necessitating the development of novel therapeutic approaches. Although biological medicines that target TNF-α (tumour necrosis factor-α) have shown clinical success in some IBD patients, a substantial proportion still fails to respond. METHODS: We designed bispecific nanobodies (BsNbs) with the ability to simultaneously target human macrophage-expressed membrane TNF-α (hmTNF-α) and IL-23. Additionally, we fused the constant region of human IgG1 Fc (hIgG1 Fc) to BsNb to create BsNb-Fc.  Our study encompassed in vitro and in vivo characterization of BsNb and BsNb-Fc. RESULTS: BsNb-Fc exhibited an improved serum half-life, targeting capability and effector function than BsNb. It's demonstrated that BsNb-Fc exhibited superior anti-inflammatory effects compared to the anti-TNF-α mAb (infliximab, IFX) combined with anti-IL-12/IL-23p40 mAb (ustekinumab, UST) by Transwell co-culture assays. Notably, in murine models of acute colitis brought on by 2,4,6-trinitrobenzene sulfonic acid（TNBS) and dextran sulphate sodium (DSS), BsNb-Fc effectively alleviated colitis severity. Additionally, BsNb-Fc outperformed the IFX&UST combination in TNBS-induced colitis, significantly reducing colon inflammation in mice with colitis produced by TNBS and DSS. CONCLUSION: These findings highlight an enhanced efficacy and improved biostability of BsNb-Fc, suggesting its potential as a promising therapeutic option for IBD patients with insufficient response to TNF-α inhibition. KEY POINTS: A bispecific nanobody (BsNb) was created to target TNF-α and IL-23p19, exhibiting high affinity and remarkable stability. BsNb-Fc inhibited the release of cytokines in CD4+T cells during co-culture experiments. BsNb-Fc effectively alleviated colitis severity in mouse model with acute colitis induced by DSS or TNBS, outperforming the IFX&UST combination.

The ERK-cPLA2-ACSL4 axis mediating M2 macrophages ferroptosis impedes mucosal healing in ulcerative colitis.

Ulcerative colitis (UC) is a chronic gastrointestinal disease that can be managed with 5-aminosalicylic acid (5-ASA), the standard treatment for UC. However, the effectiveness of 5-ASA is not always optimal. Our study revealed that despite 5-ASA treatment, cells continued to experience excessive ferroptosis, which may hinder mucosal healing in UC and limit the success of this treatment approach in achieving disease remission. We found that combining 5-ASA with the ferroptosis inhibitor Fer-1 led to a significant inhibition of ferroptosis in macrophages present in the colon tissue, along with an increase in the proportion of M2 macrophages, suggesting that targeting ferroptosis in M2 macrophages could be a potential therapeutic strategy for alleviating UC. Our study also demonstrated that M2 macrophages are more susceptible to ferroptosis compared to M1 macrophages, and this susceptibility is associated with the activated arachidonic acid (AA) metabolism pathway mediated by ERK-cPLA2-ACSL4. Additionally, we found that the expression of cPLA2 gene pla2g4a was increased in the colon of UC patients compared to healthy controls. Furthermore, targeted metabolomics analysis revealed that the combination treatment group, as opposed to the 5-ASA treatment group, exhibited the ability to modulate AA metabolism. Overall, our findings emphasize the importance of addressing macrophage ferroptosis in order to enhance macrophage anti-inflammation, improve mucosal healing, and achieve better therapeutic outcomes for patients with UC.

Ferroptosis: a promising candidate for exosome-mediated regulation in different diseases.

Ferroptosis is a newly discovered form of cell death that is featured in a wide range of diseases. Exosome therapy is a promising therapeutic option that has attracted much attention due to its low immunogenicity, low toxicity, and ability to penetrate biological barriers. In addition, emerging evidence indicates that exosomes possess the ability to modulate the progression of diverse diseases by regulating ferroptosis in damaged cells. Hence, the mechanism by which cell-derived and noncellular-derived exosomes target ferroptosis in different diseases through the system Xc-/GSH/GPX4 axis, NAD(P)H/FSP1/CoQ10 axis, iron metabolism pathway and lipid metabolism pathway associated with ferroptosis, as well as its applications in liver disease, neurological diseases, lung injury, heart injury, cancer and other diseases, are summarized here. Additionally, the role of exosome-regulated ferroptosis as an emerging repair mechanism for damaged tissues and cells is also discussed, and this is expected to be a promising treatment direction for various diseases in the future. Video Abstract.

Characterization of light-dependent rhythm of courtship vibrational signals in Nilaparvata lugens: essential involvement of cryptochrome genes.

BACKGROUND: Vibrational signal plays a crucial role in courtship communication in many insects. However, it remains unclear whether insect vibrational signals exhibit daily rhythmicity in response to changes in environmental cues. RESULTS: In this study, we observed daily rhythms of both female vibrational signals (FVS) and male vibrational signals (MVS) in the brown planthopper (BPH), Nilaparvata lugens (Stål), one of the most notorious rice pests across Asia. Notably, oscillations of FVS and MVS in paired BPHs were synchronized as part of male-female duetting interactions, displaying significant day-night rhythmicity. Furthermore, we observed light dependency of FVS emissions under different photoperiodic regimes (18 L:6 D and 6 L:18 D) and illumination intensity levels (>300 lx, 50 lx, and 25 lx). Subsequently, the potential role of circadian clock genes cryptochromes (Nlcry1 and Nlcry2) in regulating FVS daily oscillations was examined using gene knockdown via RNA interference. We observed sharp declines and disrupted rhythms in FVS frequencies when either of the Nlcrys was downregulated, with Nlcry2 knockdown showing a more prominent effect. Moreover, we recorded a novel FVS variant (with a dominant frequency of 361.76 ± 4.31 Hz) emitted by dsNlcry1-treated BPH females, which significantly diminished the impact of courtship stimuli on receptive males. CONCLUSION: We observed light-dependent daily rhythms of substrate-borne vibrational signals (SBVS) in BPH and demonstrated essential yet distinct roles of the two Nlcrys. These findings enhanced our understanding of insect SBVS and illustrated the potential of novel precision physical control strategies for disrupting mating behaviors in this rice pest. © 2023 Society of Chemical Industry.

Double n→π* Interactions with One Electron Donor: Structural and Mechanistic Insights.

Double n→π* interactions between one common electron donor of the carbonyl oxygen and two individual acceptor aldehyde/imine units are presented. The structural and mechanistic insights were revealed through a collection of experimental and computational evidence. The orientation and further energetic dependence of orbital interactions were facilely regulated by the size of cyclic urea scaffolds, the bulkiness of aldehydes/imines, and the flexibility of imine macrocycles.

Breaking through the therapeutic ceiling of inflammatory bowel disease: Dual-targeted therapies.

Emerging biologics and small-molecule drugs have changed the clinical status quo of inflammatory bowel disease (IBD). However, current treatments remain at a standstill in terms of response and remission in many cases. Accumulating evidence indicates that dual-targeted therapy (DTT) could be promising in overcoming the existing ceiling of IBD treatment. However, data on the efficacy and safety of DTT on Crohn's disease and ulcerative colitis are still limited or insufficient. Moreover, there is a lack of studies delineating the mechanisms of DTT. Given that various targeted drugs have different targets among the extensive redundant inflammatory networks, DTT could result in various outcomes. In this review, we have summarized the current data on the safety, effectiveness, and clinical development status of novel targeted drugs related to refractory IBD, and have explored the mechanism of action of therapy. We have categorized therapeutic agents into "Therapeutic Agents Targeting Cellular Signaling Pathways" and "Therapeutic Agents Targeting Leukocyte Trafficking" based on the different therapeutic targets, and also by classifying therapeutic agents targeting the cellular signaling pathways into "JAK-dependent" and "JAK-independent," and placed the existing drug combinations into 3 categories based on their mechanisms, namely, overlapping, synergistic, and complementary effects. Lastly, we have proposed the possible mechanisms of DTT to conceive a theoretical framework for clinical decision-making and further drug development and research from an IBD standpoint.

Vibrational courtship disruption of Nilaparvata lugens using artificial disruptive signals.

The brown planthopper (BPH), Nilaparvata lugens (Stål; Hemiptera: Delphacidae) is a piercing-sucking pest that causes serious damage to rice plants by sucking the phloem sap from the plants and transmitting viruses. During courtship, the BPH vibrates its abdomen to produce signals that are transmitted to rice plants through its legs. Male BPHs search, locate, and mate with female BPHs after they exchange courtship signals with each other. Currently, spraying chemical pesticides is still the primary method for controlling BPH populations in paddy fields, although this approach has led to severe environmental pollution. A physical control method based on BPH courtship disruption to reduce the mating rate is a promising strategy for cutting environmental pollution. To acquire effective courtship disruptive signals, we developed a vibration signal recording, monitoring, and playback system for BPHs. Using this system, BPH courtship signals and male competition signals were collected and analyzed to obtain their frequency spectra. Results show that the mean main vibration frequency of female courtship signals is 234 Hz and the mean pulse rate is 23 Hz. The mean main vibration and pulse frequencies of the male courtship signals are 255 Hz and 82 Hz, respectively. Besides, the mean main vibration frequency of the male competition signal is 281 Hz. Seven different forms and frequencies of artificial signals were played back to male BPHs, then the courtship and behavioral responses of male BPHs were analyzed. Results indicate that a pure tone of 225 Hz prevents the males from recognizing female courtship signals. The male reply rate fell from 95.6 to 33.3% and the mean reply delay time increased from 5.3 s to 9.1 s. The reply rates of the other six artificial signals ranged from 42.9 to 83.7%, and the mean reply delays were between 5.0 s and 9.3 s. Therefore, the courtship behavior of BPHs can be disrupted by using specific artificial disruptive signals.

An intelligent monitoring system of diseases and pests on rice canopy.

Accurate and timely surveys of rice diseases and pests are important to control them and prevent the reduction of rice yields. The current manual survey method of rice diseases and pests is time-consuming, laborious, highly subjective and difficult to trace historical data. To address these issues, we developed an intelligent monitoring system for detecting and identifying the disease and pest lesions on the rice canopy. The system mainly includes a network camera, an intelligent detection model of diseases and pests on rice canopy, a web client and a server. Each camera of the system can collect rice images in about 310 m2 of paddy fields. An improved model YOLO-Diseases and Pests Detection (YOLO-DPD) was proposed to detect three lesions of Cnaphalocrocis medinalis, Chilo suppressalis, and Ustilaginoidea virens on rice canopy. The residual feature augmentation method was used to narrow the semantic gap between different scale features of rice disease and pest images. The convolution block attention module was added into the backbone network to enhance the regional disease and pest features for suppressing the background noises. Our experiments demonstrated that the improved model YOLO-DPD could detect three species of disease and pest lesions on rice canopy at different image scales with an average precision of 92.24, 87.35 and 90.74%, respectively, and a mean average precision of 90.11%. Compared to RetinaNet, Faster R-CNN and Yolov4 models, the mean average precision of YOLO-DPD increased by 18.20, 6.98, 6.10%, respectively. The average detection time of each image is 47 ms. Our system has the advantages of unattended operation, high detection precision, objective results, and data traceability.

Intelligent Monitoring System of Migratory Pests Based on Searchlight Trap and Machine Vision.

Three species of rice migratory pests (Cnaphalocrocis medinalis, Sogatella furcifera, and Nilaparvata lugens) cause severe yield and economic losses to rice food every year. It is important that these pests are timely and accurately monitored for controlling them and ensuring food security. Insect radar is effective monitoring equipment for migratory pests flying at high altitude. But insect radar is costly and has not been widely used in fields. Searchlight trap is an economical device, which uses light to trap migratory pests at high altitude. But the trapped pests need to be manually identified and counted from a large number of non-target insects, which is inefficient and labor-intensive. In order to replace manual identification of migratory pests, we develop an intelligent monitoring system of migratory pests based on searchlight trap and machine vision. This system includes a searchlight trap based on machine vision, an automatic identification model of migratory pests, a Web client, and a cloud server. The searchlight trap attracts the high-altitude migratory insects through lights at night and kills them with the infrared heater. All trapped insects are dispersed through a multiple layers of insect conveyor belts and a revolving brush. The machine vision module collects the dispersed insect images and sends them to the cloud server through 4G network. The improved model YOLO-MPNet based on YOLOv4 and SENet channel attention mechanism is proposed to detect three species of migratory pests in the images. The results show that the model effectively improves the detection effect of three migratory pests. The precision is 94.14% for C. medinalis, 85.82% for S. furcifera, and 88.79% for N. lugens. The recall is 91.99% for C. medinalis, 82.47% for S. furcifera, and 85.00% for N. lugens. Compared with some state-of-the-art models (Faster R-CNN, YOLOv3, and YOLOv5), our model shows a low false detection and missing detection rates. The intelligent monitoring system can real-timely and automatically monitor three migratory pests instead of manually pest identification and count, which can reduce the technician workload. The trapped pest images and historical data can be visualized and traced, which provides reliable evidence for forecasting and controlling migratory pests.

Evolutionary Insights Into Microbiota Transplantation in Inflammatory Bowel Disease.

The intestinal microbiome plays an essential role in human health and disease status. So far, microbiota transplantation is considered a potential therapeutic approach for treating some chronic diseases, including inflammatory bowel disease (IBD). The diversity of gut microbiota is critical for maintaining resilience, and therefore, transplantation with numerous genetically diverse gut microbiota with metabolic flexibility and functional redundancy can effectively improve gut health than a single probiotic strain supplement. Studies have shown that natural fecal microbiota transplantation or washing microbiota transplantation can alleviate colitis and improve intestinal dysbiosis in IBD patients. However, unexpected adverse reactions caused by the complex and unclear composition of the flora limit its wider application. The evolving strain isolation technology and modifiable pre-existing strains are driving the development of microbiota transplantation. This review summarized the updating clinical and preclinical data of IBD treatments from fecal microbiota transplantation to washing microbiota transplantation, and then to artificial consortium transplantation. In addition, the factors considered for strain combination were reviewed. Furthermore, four types of artificial consortium transplant products were collected to analyze their combination and possible compatibility principles. The perspective on individualized microbiota transplantation was also discussed ultimately.