The positive impact of targeted modulating food intolerance on immune-related recurrent pregnancy loss.

BACKGROUND: It has been proven that immune disorders are one of the vital risk factors of recurrent pregnancy loss (RPL), and the presence of food intolerance seems to play an essential role in this. However, the impact of immune status induced by food intolerance on RPL has not been reported. This study utilized a targeted diet avoiding food intolerance as much as possible for each participant to investigate their effects on pregnancy outcomes in RPL patients with positive autoimmune markers. METHODS: From January 2020 to May 2021, fifty-eight patients with RPL were enrolled. They were divided into two groups based on the presence of autoantibodies: the autoantibody-positive group (AP, n = 29) and the autoantibody-negative group (AN, n = 29). Their food-specific immunoglobulin (Ig) G antibodies for 90 foods were tested using enzyme-linked immunosorbent assay (ELISA). The levels of immune parameters and the presence of gastrointestinal discomforts (diarrhea or constipation, eczema, and mouth ulcers) were recorded before and after dietary conditioning, followed by the analysis of pregnancy outcomes. RESULTS: Compared to the AN group, the patients in the AP group showed immune disorders at baseline, such as reduced levels of IL-4 and complement C3, and increased levels of IL-2 and total B cells. These parameters within the AP group were significantly improved after dietary conditioning that avoided food intolerance, while no significant changes were observed in the AN group. Patients in the AP group had significantly higher food-specific IgG antibodies for cow's milk (89.66% vs. 48.28%, p < .001), yolk (86.21% vs. 27.59%, p < .001), bamboo shoots (86.21% vs. 44.83%, p < .001) compared to those in the AN group. Additionally, gastrointestinal discomforts including diarrhea or constipation, eczema, and mouth ulcers were more common in the AP group than in the AN group. After 3-month dietary conditioning, these significantly improved characteristics were only observed in the AP group (p < .001). Finally, the baby-holding rate was higher in the AP group compared to the AN group (p < .05). CONCLUSIONS: The RPL patients in the AN group did not exhibit immune disorders, whereas those in the AP group experienced immune disorders and gastrointestinal discomforts. For patient with positive autoantibodies, dietary intervention may mitigate immune disorders and gastrointestinal discomforts, presenting a promising approach to enhance pregnancy outcomes.

Crocin's role in modulating MMP2/TIMP1 and mitigating hypoxia-induced pulmonary hypertension in mice.

To explore the molecular pathogenesis of pulmonary arterial hypertension (PAH) and identify potential therapeutic targets, we performed transcriptome sequencing of lung tissue from mice with hypoxia-induced pulmonary hypertension. Our Gene Ontology analysis revealed that "extracellular matrix organization" ranked high in the biological process category, and matrix metallopeptidases (MMPs) and other proteases also played important roles in it. Moreover, compared with those in the normoxia group, we confirmed that MMPs expression was upregulated in the hypoxia group, while the hub gene Timp1 was downregulated. Crocin, a natural MMP inhibitor, was found to reduce inflammation, decrease MMPs levels, increase Timp1 expression levels, and attenuate hypoxia-induced pulmonary hypertension in mice. In addition, analysis of the cell distribution of MMPs and Timp1 in the human lung cell atlas using single-cell RNAseq datasets revealed that MMPs and Timp1 are mainly expressed in a population of fibroblasts. Moreover, in vitro experiments revealed that crocin significantly inhibited myofibroblast proliferation, migration, and extracellular matrix deposition. Furthermore, we demonstrated that crocin inhibited TGF-ß1-induced fibroblast activation and regulated the pulmonary arterial fibroblast MMP2/TIMP1 balance by inhibiting the TGF-ß1/Smad3 signaling pathway. In summary, our results indicate that crocin attenuates hypoxia-induced pulmonary hypertension in mice by inhibiting TGF-ß1-induced myofibroblast activation.

Label-Free Direct Identification of MicroRNAs Based on a Narrow Constant-Inner-Diameter Emitter Mass Spectrometry Analysis.

MicroRNAs (miRNAs) are a class of endogenous noncoding small RNAs that play important roles in various biological processes and diseases. Direct determination of miRNAs is a cost-efficient and accurate method for analysis. Herein, we established a novel method for the analysis of miRNAs based on a narrow constant-inner-diameter mass spectrometry emitter. We utilized the gravity-assisted sleeving etching method to prepare a constant-inner-diameter mass spectrometry emitter with a capillary inner diameter of 5.5 µm, coupled it with a high-voltage power supply and a high-resolution mass spectrometer, and used it for miRNA direct detection. The method showed high sensitivity and reproducibility for the analysis of four miRNAs, with a limit of detection of 100 nmol/L (170 amol) for the Hsa-miR-1290 analysis. Compared with commercial ion sources, our method achieved higher sensitivity for miRNA detection. In addition, we analyzed the total miRNAs in the A549 cells. The result indicated that both spiked and endogenous miRNAs could be quantified with high accuracy. As a result, this method offers a promising platform for highly sensitive and accurate miRNA analysis. Furthermore, this approach can be extended to the analysis of other small oligonucleotides and holds the potential for studying clinical samples and facilitating disease diagnosis.

Activation of the alternative complement pathway modulates inflammation in thoracic aortic aneurysm/dissection.

Thoracic aortic aneurysm/dissection (TAAD) is a lethal vascular disease, and several pathological factors participate in aortic medial degeneration. We previously discovered that the complement C3a-C3aR axis in smooth muscle cells promotes the development of thoracic aortic dissection (TAD) through regulation of matrix metalloproteinase 2. However, discerning the specific complement pathway that is activated and elucidating how inflammation of the aortic wall is initiated remain unknown. We ascertained that the plasma levels of C3a and C5a were significantly elevated in patients with TAD and that the levels of C3a, C4a, and C5a were higher in acute TAD than in chronic TAD. We also confirmed the activation of the complement in a TAD mouse model. Subsequently, knocking out Cfb (Cfb) or C4 in mice with TAD revealed that the alternative pathway and Cfb played a significant role in the TAD process. Activation of the alternative pathway led to generation of the anaphylatoxins C3a and C5a, and knocking out their receptors reduced the recruitment of inflammatory cells to the aortic wall. Moreover, we used serum from wild-type mice or recombinant mice Cfb as an exogenous source of Cfb to treat Cfb KO mice and observed that it exacerbated the onset and rupture of TAD. Finally, we knocked out Cfb in the FBN1C1041G/+ Marfan-syndrome mice and showed that the occurrence of TAA was reduced. In summary, the alternative complement pathway promoted the development of TAAD by recruiting infiltrating inflammatory cells. Targeting the alternative pathway may thus constitute a strategy for preventing the development of TAAD.NEW & NOTEWORTHY The alternative complement pathway promoted the development of TAAD by recruiting infiltrating inflammatory cells. Targeting the alternative pathway may thus constitute a strategy for preventing the development of TAAD.

Zein/hyaluronic acid nanoparticle stabilized Pickering emulsion for astaxanthin encapsulation.

Pickering emulsions have attracted considerable attention owing to the stability and functionality. In this study, zein/hyaluronic acid (ZH) nanoparticles were prepared and applied for stabilizing astaxanthin encapsulated Pickering emulsions. By non-covalent interaction between Zein and hyaluronic acid (HA), the conformation of zein changed and therefore improved the wettability of ZH nanoparticles. Unlike the spherical zein nanoparticles, ZH nanoparticles possessed a cross-linked structure with rough surface. Confocal laser scanning microscopy indicated that the nanoparticles accumulated at the oil-water interface. The Pickering emulsion stabilized by ZH nanoparticles exhibited high viscoelasticity and a solid-like behavior, as well as excellent stability during the storage. In vitro digestion results revealed that the presence of HA coating prevented the emulsion from pepsin hydrolysis and achieved efficient delivery of astaxanthin. This work confirmed that Pickering emulsion stabilized by ZH nanoparticles could be used as an effective deliver system for bioactive substances.

Quantitative Analysis of Nanoplastics in Single Cells by Subcellular Chromatography.

The accumulation and spatial distribution of intracellular nanoplastic particles provide useful information about their spatiotemporal toxicological effects mediated by the physicochemical parameters of nanoplastics in living cells. In this study, a sample injection-transfer method was designed with an accuracy of up to femtoliters to attoliters to match the volume required for ultranarrow-bore open-tubular liquid chromatography. The separation and concentration quantification of mixed polystyrenes in different regions in living cells were achieved by directly transferring picoliter/femtoliter volumes of intracellular cytoplasm to an ultranarrow-bore open-tubular chromatographic column. The measurement of pollutant concentration in different areas of a small-volume target (single cell) was realized. This method is expected to be used in the qualitative and quantitative analyses of complex, mixed, and label-free nanoplastics (a few nm in size) in the subregions of living cells.

Current status of diagnosis and treatment of pulmonary hypertension in Chinese tertiary hospitals: A nationwide survey.

We intended to evaluate the diagnosis and treatment status of pulmonary hypertension (PH) in China and provide the basis for the design of the Chinese PH centers system. A questionnaire survey was conducted by sampling from Chinese Class A tertiary hospitals that have carried out the clinical work of PH, including the composition of PH clinical team, the current application of examinations related to PH diagnosis, the availability of PAH-specific medicine and the implementation of PH-related intervention and surgery. A total of 44 valid questionnaires from 20 provinces were collected in this survey. In the vast majority of centers (83.33%, n = 35), pulmonary artery catheterization was routinely performed under X-ray guidance. In 19.05% (n = 8) of centers, pressure measurements were determined at the right time (the end of normal expiration). Only 73.81% (n = 31) centers have carried out acute vasoreactivity testing. Prostacyclin analogues and prostaglandin receptor agonists were just prescribed in 45.45% (n = 20) of the centers. 19 centers (43.18%) were capable of performing balloon pulmonary angioplasty (BPA) and pulmonary endarterectomy (PEA), while 25% (n = 11) were able to perform BPA, PEA, and lung transplantation. There was no significant difference in the diagnosis and treatment of PH between economic regions. The majority of Chinese tertiary hospitals were well equipped with the corresponding personnel, examinations and medicines related to PH, but the standardization and specialization of the management of PH need to be strengthened.

A comparative study on the diagnostic efficacy of different diagnostic criteria for exercise pulmonary hypertension.

BACKGROUND: Exercise pulmonary hypertension (ePH) has three common diagnostic criteria: the mean pulmonary artery pressure (mPAP) > 30 mmHg and total pulmonary resistance (TPR) at peak exercise >3 Wood units ("Joint criteria"), the mPAP/cardiac output (CO) slope of the two-point measurement (ΔmPAP/ΔCO) > 3 mmHg/L/min ("Two-point criteria"), and the mPAP/CO slope of the multi-point data >3 mmHg/L/min ("Multi-point criteria"). We compared the diagnostic efficacy of these criteria, which remain controversial. METHODS: Following resting right heart catheterization (RHC), all patients underwent exercise RHC (eRHC). The patients were divided into different ePH and non-exercise pulmonary hypertension (nPH) groups according to the above criteria. Joint criteria were used as the reference to compare the other two, namely diagnostic concordance, sensitivity and specificity. We conducted further analysis to determine the correlation between different diagnostic criteria grouping and the clinical severity of PH. RESULTS: Thirty-three patients with mPAPrest ≤ 20 mmHg were enrolled. a) Diagnostic concordance, sensitivity and specificity: compared with Joint criteria, the diagnostic concordances of Two-point criteria and Multi-point criteria were 78.8% (κ = 0.570, P < 0.01) and 90.9% (κ = 0.818, P < 0.01), respectively; the sensitivity of Two-point criteria was high (100%), but the specificity was poor (56.3%); however, Multi-point criteria exhibited higher sensitivity (94.1%) and specificity (87.5%). b) Clinically relevant analysis: a significant difference was observed in several clinical severity indicators between ePH and nPH patients according to Multi-point criteria grouping(all P < 0.05). CONCLUSION: Multi-point criteria are more clinically relevant and provide better diagnostic efficiency.

Single-Cell Metabolomics-Based Strategy for Studying the Mechanisms of Drug Action.

Studying the mechanisms of drug antitumor activity at the single-cell level can provide information about the responses of cell subpopulations to drug therapy, which is essential for the accurate treatment of cancer. Due to the small size of single cells and the low contents of metabolites, metabolomics-based approaches to studying the mechanisms of drug action at the single-cell level are lacking. Herein, we develop a label-free platform for studying the mechanisms of drug action based on single-cell metabolomics (sMDA-scM) by integrating intact living-cell electro-launching ionization mass spectrometry (ILCEI-MS) with metabolomics analysis. Using this platform, we reveal that non-small-cell lung cancer (NSCLC) cells treated by gefitinib can be clustered into two cell subpopulations with different metabolic responses. The glutathione metabolic pathway of the subpopulation containing 14.4% of the cells is not significantly affected by gefitinib, exhibiting certain resistance characteristics. The presence of these cells masked the judgment of whether cysteine and methionine metabolic pathway was remarkably influenced in the analysis of overall average results, revealing the heterogeneity of the response of single NSCLC cells to gefitinib treatment. The findings provide a basis for evaluating the early therapeutic effects of clinical medicines and insights for overcoming drug resistance in NSCLC subpopulations.

Progressive interstitial lung disease in hypomyopathic dermatomyositis in the COVID-19 pandemic: A case report.

BACKGROUND: Clinically amyopathic dermatomyositis (CADM) is characterized by typical skin lesions with no (amyopathic) or subclinical (hypomyopathic) evidence of muscle involvement. Patients with CADM may also develop rapidly progressive interstitial lung disease (ILD), and have a poor prognosis. However, the diagnosis of rapidly progressive ILD faces a challenge during the severe acute respiratory syndrome coronavirus 2 pandemic. Severe acute respiratory syndrome and ground-glass attenuation on a chest computed tomography scan are the presenting features in both conditions. CASE PRESENTATION: A 45-year-old woman with amyopathic dermatomyositis had acute onset of fever and dyspnea in February 2020. She had abnormal lung findings on CT scan. Polymerase chain reaction testing for SARS-CoV-2 was not available at that time. Chest CT revealed non-specific manifestations that could be either the signs of ILD or SARS-CoV-2 infection. Antiviral therapy was initiated with oseltamivir. Three days later, she had erythema on face, palm, and back. The ratio of lactate dehydrogenase (LDH) isoenzyme 3 to total LDH was elevated. The ratio of LDH isoenzyme 1 to total LDH was declined. Therefore, she was transferred to the rheumatology ward for further treatment. However, she died from respiratory failure 2 weeks later. CONCLUSIONS: We speculate that the altered LDH isoenzyme pattern may be an early biomarker for co-occurrence of CADM and ILD.

VO2-assisted multifunctional metamaterial for polarization conversion and asymmetric transmission.

A VO2-assisted temperature-controlled multifunctional metamaterial polarization converter with large asymmetric transmission (AT) is proposed by introducing a gold-VO2 grating. The converter can be switched between reflection mode and transmission mode by controlling the phase transition. When VO2 is in the metallic state, the converter works in reflection mode, converting the incident forward/backward linearly/circularly polarized waves into the cross-polarized waves, and the broadband polarization conversion rates (PCRs) can reach 90% with relative bandwidth of up to 91.1% and 87.5%, respectively; when VO2 is in the insulating state, the converter shows giant AT effect for circularly polarized waves at 0.64 THz and 1.28 THz. The multifunctional polarization converter holds great potential in the fields of communication and imaging, which provides a new way to design optical devices such as polarizers, isolators.

A novel pyroptosis related genes signature for predicting prognosis and estimating tumor immune microenvironment in lung adenocarcinoma.

Background: Pyroptosis is a newly found form of programmed cell death, accompanied by inflammatory response as well as immune response. Here, the specific function and prognosis predictive of pyroptosis-related genes (PRGs) were systematically explored in lung adenocarcinoma (LUAD). Methods: The gene expression data and corresponding clinical information of LUAD patients were obtained from The Cancer Genome Atlas (TCGA), and the expression level of PRGs was identified between normal and tumor tissues. Furthermore, univariate Cox proportional hazards regression was conducted to filter the PRGs related to overall survival, and least absolute shrinkage and selection operator (LASSO) regression was subsequent employed to establish the PRGs risk model. Besides, the correlation of risk score with patients' clinical features, tumor mutational burden (TMB) as well as tumor microenvironment (TME) was also investigated. Results: A total of 5 PRGs (NLRC4, NLRP1, NLRP3, NOD1, PLCG1, and BAK1) was used to establish the risk prognostic model. According the median value of risk score, all the patients were classified into low- and high-risk score group. Kaplan-Meier analysis indicted that the LUAD patients in low-risk group exhibited a better survival outcome compared the patients in high-risk group (P<0.001). After adjusting for age, gender, and clinical stage, the risk score was also considered as and independent risk factor affecting the overall survival of LUAD patients (HR =2.949, 95% CI: 1.762-4.937). Moreover, low-risk score group exhibited a higher Immune score and lower Tumor purity compared with high-risk score group. ssGSEA results proved that the enrichment scores of most immune cells and immune related signal pathway in low-risk score group was significant higher than that in high-risk score group. In addition, the PRGs risk score was also positive correlated with TMB in LUAD tissues. Conclusions: In this study, a novel prognostic model based on PRGs was constructed and used to predict the survival outcome of LUAD patients. In addition, the PRGs risk signature was also associated with TMB and anti-tumor immune environment. The induction of pyroptosis inside tumors might be considered a potential strategy in cancer treatments.

Identification of Signal Pathways and Hub Genes of Pulmonary Arterial Hypertension by Bioinformatic Analysis.

Pulmonary arterial hypertension (PAH) is a progressive and complex pulmonary vascular disease with poor prognosis. The aim of this study was to provide a new understanding of the pathogenesis of disease and potential treatment targets for patients with PAH based on multiple-microarray analysis.Two microarray datasets (GSE53408 and GSE113439) downloaded from the Gene Expression Omnibus (GEO) database were analysed. All the raw data were processed by R, and differentially expressed genes (DEGs) were screened out by the "limma" package. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses were performed and visualized by R and Cytoscape software. Protein-protein interactions (PPI) of DEGs were analysed based on the NetworkAnalyst online tool. A total of 442 upregulated DEGs and 84 downregulated DEGs were identified. GO enrichment analysis showed that these DEGs were mainly enriched in mitotic nuclear division, organelle fission, chromosome segregation, nuclear division, and sister chromatid segregation. Significant KEGG pathway enrichment included ribosome biogenesis in eukaryotes, RNA transport, proteoglycans in cancer, dilated cardiomyopathy, rheumatoid arthritis, vascular smooth muscle contraction, focal adhesion, regulation of the actin cytoskeleton, and hypertrophic cardiomyopathy. The PPI network identified 10 hub genes including HSP90AA1, CDC5L, MDM2, LRRK2, CFTR, IQGAP1, CAND1, TOP2A, DDX21, and HIF1A. We elucidated potential biomarkers and therapeutic targets for PAH by bioinformatic analysis, which provides a theoretical basis for future study.

Intact living-cell electrolaunching ionization mass spectrometry for single-cell metabolomics.

While single-cell mass spectrometry can reveal cellular heterogeneity and the molecular mechanisms of intracellular biochemical reactions, its application is limited by the insufficient detection sensitivity resulting from matrix interference and sample dilution. Herein, we propose an intact living-cell electrolaunching ionization mass spectrometry (ILCEI-MS) method. A capillary emitter with a narrow-bore, constant-inner-diameter ensures that the entire living cell enters the MS ion-transfer tube. Inlet ionization improves sample utilization, and no solvent is required, preventing sample dilution and matrix interference. Based on these features, the detection sensitivity is greatly improved, and the average signal-to-noise (S/N) ratio is about 20 : 1 of single-cell peaks in the TIC of ILCEI-MS. A high detection throughput of 51 cells per min was achieved by ILCEI-MS for the single-cell metabolic profiling of multiple cell lines, and 368 cellular metabolites were identified. Further, more than 4000 primary single cells digested from the fresh multi-organ tissues of mice were detected by ILCEI-MS, demonstrating its applicability and reliability.

Broadband Design of Midinfrared Chiral Metamaterials Based on the Indium Tin Oxide Conical Helix.

This paper proposed a periodic midinfrared broadband chiral structure composed of indium tin oxide (ITO) conical helix. The simulation results show that the structure achieves a good broadband CD in the midinfrared. At the wavelength of 9.2 µm, the maximum value of CD is 0.55, the FWHM (full width half maximum) of CD is 7.2 µm, and the wavelength range is from 5.7 µm to 12.9 µm. The simulation results also show that compared with the traditional uniform spiral structure with the same radius, the conical spiral structure can achieve better broadband circular dichroism in the midinfrared band. This provides a new idea for the design of broadband polarization state control devices in the midinfrared band.

Analytical methods for obtaining binding parameters of drug-protein interactions: A review.

The study of drug-protein interactions can reveal the corresponding binding mechanisms, providing valuable information for the early phase drug development and development of new drugs. This article reviews the methods used for obtaining the binding parameters of drug-protein systems. The methods include equilibrium dialysis, high-performance affinity chromatography, capillary electrophoresis, spectroscopy, calorimetry, competition and displacement, mass spectrometry, fluorescence resonance energy transfer, and thermal stability shift analysis. Relevant parameters include the association constant, number of binding sites, thermodynamic properties, binding force types, binding site types, binding distances, changes in protein conformation, and changes in protein stability. In addition, the review also summarizes the principles, advantages, and limitations of each method in detail. The comparison of parameter information can not only guide method selection but also provide valuable reference information for in-depth exploration of drug-protein interaction mechanisms.

Small proline-rich protein 1A promotes lung adenocarcinoma progression and indicates unfavorable clinical outcomes.

Small proline-rich protein 1A (SPRR1A) plays a critical role in regulating squamous cell differentiation. SPRR1A overexpression was reported to be closely related to the progression of some tumors, such as gastric cancer and colon cancer. However, the function of SPRR1A in lung adenocarcinoma (LUAD) has not been elucidated. Here, we first examined the expression pattern of SPRR1A in LUAD tissues, which indicated that the SPRR1A expression level was significantly elevated in LUAD tissues compared with normal lung tissues. High expression of SPRR1A was closely related to larger tumor size. LUAD patients with higher SPRR1A expression had poorer overall survival and SPRR1A was identified as an independent unfavorable prognosis factor. In addition, the effects of SPRR1A on lung cancer cells were tested through cellular experiments and the result demonstrated that knockdown of SPRR1A can suppress the proliferation and invasion capacities of tumor cells, while overexpressing SPRR1A exerted opposite effects. Finally, our findings were substantiated by the data obtained from in vivo xenografts using a mice model. In conclusion, LUAD patients with higher SPRR1A expression were more predisposed to poorer clinical outcomes and unfavorable prognoses, indicating the potential role of SPRR1A as a novel clinical biomarker and therapeutic target.

Super-resolution scanning imaging based on metal-dielectric composite metamaterials.

We propose super-resolution scanning imaging by using a metamaterial composed of a silver-silicon dioxide composite covered by a layer of chromium containing one slit and a silicon dioxide substrate. By simulating a distribution of energy flow in the metamaterial for an H-polarized wave, we find that the output beam exhibits focusing accompanied with good directional radiation, which is able to be designed as a super-resolution scanning probe. We also demonstrate numerically super-resolution imaging by scanning our designed metamaterial over a sub-wavelength object.

Investigation of metformin hydrochloride-bovine serum albumin interaction by narrow-bore capillary zone electrophoresis.

A narrow-bore capillary (inner diameter = 2 µm) zone electrophoresis method was developed to study the interaction between metformin hydrochloride and bovine serum albumin. Free metformin hydrochloride, free bovine serum albumin, and their complex were completely separated in a volume of tens of picoliters. The association constant (Ka = 1.04 × 103 L mol-1) and the number of binding sites (n = 0.9789) were then obtained.

High-Throughput NanoBiT-Based Screening for Inhibitors of HIV-1 Vpu and Host BST-2 Protein Interaction.

Bone marrow stromal cell antigen 2 (BST-2), also known as CD317 or tetherin, has been identified as a host restriction factor that suppresses the release of enveloped viruses from host cells by physically tethering viral particles to the cell surface; however, this host defense can be subverted by multiple viruses. For example, human immunodeficiency virus (HIV)-1 encodes a specific accessory protein, viral protein U (Vpu), to counteract BST-2 by binding to it and directing its lysosomal degradation. Thus, blocking the interaction between Vpu and BST-2 will provide a promising strategy for anti-HIV therapy. Here, we report a NanoLuc Binary Technology (NanoBiT)-based high-throughput screening assay to detect inhibitors that disrupt the Vpu-BST-2 interaction. Out of more than 1000 compounds screened, four inhibitors were identified with strong activity at nontoxic concentrations. In subsequent cell-based BST-2 degradation assays, inhibitor Y-39983 HCl restored the cell-surface and total cellular level of BST-2 in the presence of Vpu. Furthermore, the Vpu-mediated enhancement of pesudotyped viral particle production was inhibited by Y-39983 HCl. Our findings indicate that our newly developed assay can be used for the discovery of potential antiviral molecules with novel mechanisms of action.