Inhibition of porcine reproductive and respiratory syndrome virus replication by rifampicin in vitro.

Porcine reproductive and respiratory syndrome virus (PRRSV) continues to cause significant economic losses to the global swine industry, yet effective prevention and control measures remain elusive. The development of novel antivirals is thus urgently needed. Rifampicin (RFP), a semisynthetic derivative of rifamycin, has been previously reported to inhibit the replication of certain mammalian DNA viruses as well as RNA viruses. In this study, we unveil RFP as a potent inhibitor of PRRSV both in Marc-145 cells (half-maximal inhibitory concentration 61.26 µM) and porcine alveolar macrophages (half-maximal inhibitory concentration 53.09 µM). The inhibitory effect of RFP occurred during viral replication rather than binding, internalization and release. We also demonstrated that RFP inhibits PRRSV proteins production in the early stage of infection, without inhibiting host protein synthesis. Moreover, RFP effectively restricted porcine epidemic diarrhea virus (PEDV) and porcine enteric alphacoronavirus (PEAV) infection in Vero cells. In summary, these findings indicate the promising potential of RFP as a therapeutic agent for PRRSV, PEDV and PEAV infection in pig farms.

Predicting sequenced dental treatment plans from electronic dental records using deep learning.

BACKGROUND: Designing appropriate clinical dental treatment plans is an urgent need because a growing number of dental patients are suffering from partial edentulism with the population getting older. OBJECTIVES: The aim of this study is to predict sequential treatment plans from electronic dental records. METHODS: We construct a clinical decision support model, MultiTP, explores the unique topology of teeth information and the variation of complicated treatments, integrates deep learning models (convolutional neural network and recurrent neural network) adaptively, and embeds the attention mechanism to produce optimal treatment plans. RESULTS: MultiTP shows its promising performance with an AUC of 0.9079 and an F score of 0.8472 over five treatment plans. The interpretability analysis also indicates its capability in mining clinical knowledge from the textual data. CONCLUSIONS: MultiTP's novel problem formulation, neural network framework, and interpretability analysis techniques allow for broad applications of deep learning in dental healthcare, providing valuable support for predicting dental treatment plans in the clinic and benefiting dental patients. CLINICAL IMPLICATIONS: The MultiTP is an efficient tool that can be implemented in clinical practice and integrated into the existing EDR system. By predicting treatment plans for partial edentulism, the model will help dentists improve their clinical decisions.

The pig pangenome provides insights into the roles of coding structural variations in genetic diversity and adaptation.

Structural variations have emerged as an important driving force for genome evolution and phenotypic variation in various organisms, yet their contributions to genetic diversity and adaptation in domesticated animals remain largely unknown. Here we constructed a pangenome based on 250 sequenced individuals from 32 pig breeds in Eurasia and systematically characterized coding sequence presence/absence variations (PAVs) within pigs. We identified 308.3-Mb nonreference sequences and 3438 novel genes absent from the current reference genome. Gene PAV analysis showed that 16.8% of the genes in the pangene catalog undergo PAV. A number of newly identified dispensable genes showed close associations with adaptation. For instance, several novel swine leukocyte antigen (SLA) genes discovered in nonreference sequences potentially participate in immune responses to productive and respiratory syndrome virus (PRRSV) infection. We delineated previously unidentified features of the pig mobilome that contained 490,480 transposable element insertion polymorphisms (TIPs) resulting from recent mobilization of 970 TE families, and investigated their population dynamics along with influences on population differentiation and gene expression. In addition, several candidate adaptive TE insertions were detected to be co-opted into genes responsible for responses to hypoxia, skeletal development, regulation of heart contraction, and neuronal cell development, likely contributing to local adaptation of Tibetan wild boars. These findings enhance our understanding on hidden layers of the genetic diversity in pigs and provide novel insights into the role of SVs in the evolutionary adaptation of mammals.

PRRSV Infection Induces Gasdermin D-Driven Pyroptosis of Porcine Alveolar Macrophages through NLRP3 Inflammasome Activation.

For more than 3 decades, mounting evidence has associated porcine reproductive and respiratory syndrome virus (PRRSV) infection with late-term abortions and stillbirths in sows and respiratory disease in piglets, causing enormous economic losses to the global swine industry. However, to date, the underlying mechanisms of PRRSV-triggered cell death have not been well clarified, especially in the pulmonary inflammatory injury characterized by the massive release of pro-inflammatory factors. Here, we demonstrated that PRRSV infection triggered gasdermin D-mediated host pyroptosis in vitro and in vivo. Mechanistically, PRRSV infection triggered disassembly of the trans-Golgi network (TGN); the dispersed TGN then acted as a scaffold for NLRP3 activation through phosphatidylinositol-4-phosphate. In addition, PRRSV replication-transcription complex (RTC) formation stimulated TGN dispersion and pyroptotic cell death. Furthermore, our results indicated that TMEM41B, an endoplasmic reticulum (ER)-resident host protein, functioned as a crucial host factor in the formation of PRRSV RTC, which is surrounded by the intermediate filament network. Collectively, these findings uncover new insights into clinical features as previously unrecognized mechanisms for PRRSV-induced pathological effects, which may be conducive to providing treatment options for PRRSV-associated diseases and may be conserved during infection by other highly pathogenic viruses. IMPORTANCE Porcine reproductive and respiratory syndrome virus (PRRSV) is one of the pathogens responsible for major economic losses in the global swine industry. Characterizing the detailed process by which PRRSV induces cell death pathways will help us better understand viral pathogenesis and provide implications for therapeutic intervention against PRRSV. Here, we showed that PRRSV infection induces GSDMD-driven host pyroptosis and IL-1ß secretion through NOD-, LRR- and pyrin domain-containing protein 3 (NLRP3) inflammasome activation in vitro and in vivo. Furthermore, the molecular mechanisms of PRRSV-induced NLRP3 inflammasome activation and pyroptosis are elucidated here. The dispersed trans-Golgi network (TGN) induced by PRRSV serves as a scaffold for NLRP3 aggregation into multiple puncta via phosphatidylinositol 4-phosphate (PtdIns4P). Moreover, the formation of PRRSV replication-transcription complex is essential for TGN dispersion and host pyroptosis. This research advances our understanding of the PRRSV-mediated inflammatory response and cell death pathways, paving the way for the development of effective treatments for PRRSV diseases.

Tandutinib (MLN518) reverses multidrug resistance by inhibiting the efflux activity of the multidrug resistance protein 7 (ABCC10).

It is well established that ATP-binding cassette (ABC) transporter-mediated multidrug resistance (MDR) is one of the major mechanisms that causes resistance to antineoplastic drugs in cancer cells. ABC transporters can significantly decrease the intracellular concentration of antineoplastic drugs by increasing their efflux, thereby lowering their cytotoxic activity. One of these transporters, the multidrug resistance protein 7 (MRP7/ABCC10), has already been shown to produce resistance to antineoplastic drugs by increasing the efflux of the drugs. In the present study, we investigated whether tandutinib, an FMS-like tyrosine kinase 3 (FLT3) inhibitor, has the potential to reverse MRP7-mediated MDR. Our results revealed that tandutinib significantly enhanced the sensitivity of MRP7-transfected HEK293 cells to the 2 established MRP7 substrates, paclitaxel and vincristine, whereas there was less or no effect on the control vector-transfected HEK293 cells. [³H]-paclitaxel accumulation and efflux studies demonstrated that tandutinib increased the intracellular accumulation of [³H]-paclitaxel and inhibited the efflux of [³H]-paclitaxel from HEK-MRP7 cells. In addition, western blot analysis showed that tandutinib did not significantly affect MRP7 expression. Thus, we conclude that the FLT3 inhibitor tandutinib can reverse MRP7-mediated MDR through inhibition of the drug efflux function and may have potential to be used clinically in combination therapy for cancer patients.

Measuring thoracic excursion: reliability of the cloth tape measure technique.

OBJECTIVE: To assess the reliability of using a cloth tape measure to determine thoracic respiratory excursion as a measurement of chest expansion or mobility. METHODS: Physicians and residents experienced in osteopathic manipulative treatment measured thoracic excursion with a cloth tape measure held around the circumference of healthy male subjects' chests at two levels. Upper thoracic excursion measurements were taken at the level of the fifth thoracic spinous process and the third intercostal space at the midclavicular line. Lower thoracic excursion measurements were taken at the level of the 10th thoracic spinous process and the xiphoid process. At peak inhalation and exhalation, three examiners measured thoracic excursion at both levels. In the first session (n=5), examiners measured the same subject inhalation and exhalation. In the second session (n=4), examiners measured separate respiratory cycles. For each session, interexaminer intraclass correlation coefficients (ICCs) were calculated for thoracic excursion, inhalation, and exhalation in the upper and lower positions using a two-way random-effects analysis of variance model. RESULTS: Intraclass correlation coefficients for thoracic excursion ranged from 0.81 to 0.91 (95% confidence interval, 0.69-0.99) at both measurement levels in both sessions. When inhalation and exhalation were considered separately, interexaminer ICCs were 0.99 and greater. Standard deviations for measurements of each subject's thoracic excursion at both levels ranged from 0.5 cm to 0.8 cm with a mean of 0.6 cm. CONCLUSION: The method of using a tape measure to assess thoracic excursion was highly reliable in men, resulting in ICCs of substantial reliability. The SDs at each level of measurement indicate that this method may be most useful in measuring changes in thoracic excursion that are expected to be 0.6 cm or greater.