CRISPR/Cas9-based toolkit for rapid marker recycling and combinatorial libraries in Komagataella phaffii.

Komagataella phaffii, a nonconventional yeast, is increasingly attractive to researchers owing to its posttranslational modification ability, strict methanol regulatory mechanism, and lack of Crabtree effect. Although CRISPR-based gene editing systems have been established in K. phaffii, there are still some inadequacies compared to the model organism Saccharomyces cerevisiae. In this study, a redesigned gRNA plasmid carrying red and green fluorescent proteins facilitated plasmid construction and marker recycling, respectively, making marker recycling more convenient and reliable. Subsequently, based on the knockdown of Ku70 and DNA ligase IV, we experimented with integrating multiple DNA fragments at a single locus. A 26.5-kb-long DNA fragment divided into 11 expression cassettes for lycopene synthesis could be successfully integrated into a single locus at one time with a success rate of 57%. A 27-kb-long DNA fragment could also be precisely knocked out with a 50% positive rate in K. phaffii by introducing two DSBs simultaneously. Finally, to explore the feasibility of rapidly balancing the expression intensity of multiple genes in a metabolic pathway, a yeast combinatorial library was successfully constructed in K. phaffii using lycopene as an indicator, and an optimal combination of the metabolic pathway was identified by screening, with a yield titer of up to 182.73 mg/L in shake flask fermentation. KEY POINTS: • Rapid marker recycling based on the visualization of a green fluorescent protein • One-step multifragment integration and large fragment knockout in the genome • A random assembly of multiple DNA elements to create yeast libraries in K. phaffii.

Developmental role of macrophages modeled in human pluripotent stem cell-derived intestinal tissue.

Macrophages populate the embryo early in gestation, but their role in development is not well defined. In particular, specification and function of macrophages in intestinal development remain little explored. To study this event in the human developmental context, we derived and combined human intestinal organoid and macrophages from pluripotent stem cells. Macrophages migrate into the organoid, proliferate, and occupy the emerging microanatomical niches of epithelial crypts and ganglia. They also acquire a transcriptomic profile similar to that of fetal intestinal macrophages and display tissue macrophage behaviors, such as recruitment to tissue injury. Using this model, we show that macrophages reduce glycolysis in mesenchymal cells and limit tissue growth without affecting tissue architecture, in contrast to the pro-growth effect of enteric neurons. In short, we engineered an intestinal tissue model populated with macrophages, and we suggest that resident macrophages contribute to the regulation of metabolism and growth of the developing intestine.

Quantifying global colonization pressures of alien vertebrates from wildlife trade.

The global trade in live wildlife elevates the risk of biological invasions by increasing colonization pressure (the number of alien species introduced to an area). Yet, our understanding of species traded as aliens remains limited. We created a comprehensive global database on live terrestrial vertebrate trade and use it to investigate the number of traded alien species, and correlates of establishment richness for aliens. We identify 7,780 species involved in this trade globally. Approximately 85.7% of these species are traded as aliens, and 12.2% of aliens establish populations. Countries with greater trading power, higher incomes, and larger human populations import more alien species. These countries, along with island nations, emerge as hotspots for establishment richness of aliens. Colonization pressure and insularity consistently promote establishment richness across countries, while socio-economic factors impact specific taxa. Governments must prioritize policies to mitigate the release or escape of traded animals and protect global biosecurity.

KLRC1 knockout overcomes HLA-E-mediated inhibition and improves NK cell antitumor activity against solid tumors.

Introduction: Natural Killer (NK) cells hold the potential to shift cell therapy from a complex autologous option to a universal off-the-shelf one. Although NK cells have demonstrated efficacy and safety in the treatment of leukemia, the limited efficacy of NK cell-based immunotherapies against solid tumors still represents a major hurdle. In the immunosuppressive tumor microenvironment (TME), inhibitory interactions between cancer and immune cells impair antitumoral immunity. KLRC1 gene encodes the NK cell inhibitory receptor NKG2A, which is a potent NK cell immune checkpoint. NKG2A specifically binds HLA-E, a non-classical HLA class I molecule frequently overexpressed in tumors, leading to the transmission of inhibitory signals that strongly impair NK cell function. Methods: To restore NK cell cytotoxicity against HLA-E+ tumors, we have targeted the NKG2A/HLA-E immune checkpoint by using a CRISPR-mediated KLRC1 gene editing. Results: KLRC1 knockout resulted in a reduction of 81% of NKG2A+ cell frequency in ex vivo expanded human NK cells post-cell sorting. In vitro, the overexpression of HLA-E by tumor cells significantly inhibited wild-type (WT) NK cell cytotoxicity with p-values ranging from 0.0071 to 0.0473 depending on tumor cell lines. In contrast, KLRC1 KO NK cells exhibited significantly higher cytotoxicity when compared to WT NK cells against four different HLA-E+ solid tumor cell lines, with p-values ranging from<0.0001 to 0.0154. Interestingly, a proportion of 43.5% to 60.2% of NKG2A- NK cells within the edited NK cell population was sufficient to reverse at its maximum the HLA-E-mediated inhibition of NK cell cytotoxicity. The expression of the activating receptor NKG2C was increased in KLRC1 KO NK cells and contributed to the improved NK cell cytotoxicity against HLA-E+ tumors. In vivo, the adoptive transfer of human KLRC1 KO NK cells significantly delayed tumor progression and increased survival in a xenogeneic mouse model of HLA-E+ metastatic breast cancer, as compared to WT NK cells (p = 0.0015). Conclusions: Our results demonstrate that KLRC1 knockout is an effective strategy to improve NK cell antitumor activity against HLA-E+ tumors and could be applied in the development of NK cell therapy for solid tumors.

Effects of Fermentation on Bioactivity and the Composition of Polyphenols Contained in Polyphenol-Rich Foods: A Review.

Polyphenols, as common components with various functional activities in plants, have become a research hotspot. However, researchers have found that the bioavailability and bioactivity of plant polyphenols is generally low because they are usually in the form of tannins, anthocyanins and glycosides. Polyphenol-rich fermented foods (PFFs) are reported to have better bioavailability and bioactivity than polyphenol-rich foods, because polyphenols are used as substrates during food fermentation and are hydrolyzed into smaller phenolic compounds (such as quercetin, kaempferol, gallic acid, ellagic acid, etc.) with higher bioactivity and bioavailability by polyphenol-associated enzymes (PAEs, e.g., tannases, esterases, phenolic acid decarboxylases and glycosidases). Biotransformation pathways of different polyphenols by PAEs secreted by different microorganisms are different. Meanwhile, polyphenols could also promote the growth of beneficial bacteria during the fermentation process while inhibiting the growth of pathogenic bacteria. Therefore, during the fermentation of PFFs, there must be an interactive relationship between polyphenols and microorganisms. The present study is an integration and analysis of the interaction mechanism between PFFs and microorganisms and is systematically elaborated. The present study will provide some new insights to explore the bioavailability and bioactivity of polyphenol-rich foods and greater exploitation of the availability of functional components (such as polyphenols) in plant-derived foods.

Interaction of glycine with Li+ in the (H2O)n (n = 0-8) clusters.

CONTEXT: We investigated the interaction between glycine and Li+ in water environment based on the Gly·Li+(H2O)n (n = 0-8) cluster. Our study shows that for Gly·Li+, Li+ binds to both carbonyl oxygen and amino nitrogen to form a bidentate structure, and the first three water molecules preferentially interact with Li+. For n = 0-5, the complexes of Gly·Li+(H2O)n exist in neutral form, and when the water number reached 6, the complex can coexist in neutral and zwitterionic form, then zwitterionic structures are dominant for n = 7, 8. The analyses by RDG, AIM, and ESP in conjunction with the calculated interaction energies show that the interaction between Li+ and Gly decreases gradually with the water molecules involved successively from n = 1 to 6 and then increases for n = 7-8. Additionally, the infrared spectra of Gly·Li+(H2O)n (n = 0-8) are also calculated. METHODS: The initial structures were optimized using Gaussian 09 program package in B3LYP-D3 (BJ)/6-311G(d, p) method, and the frequency was calculated with 6-311 + G(2d, p) basis set. GaussView5.0.9 was used to view simulation infrared spectra. The noncovalent interaction method (NCl), energy decomposition (EDA), atoms in molecules (AIM) analysis, and electrostatic potential (ESP) analyses were conducted using Multiwfn software to gain a deeper understanding of the interaction properties of Gly, Li+, and water.

Immobilized glucoamylase based on ZIF-8: Preparation, response surface optimization, characterization.

The glucoamylase@ZIF-8 was prepared using ZIF-8 material as the carrier in this study. The preparation process was optimized by response surface methodology, and the stability of glucoamylase@ZIF-8 was determined. The material was characterized by scanning electron microscopy, X-ray diffraction, and Fourier transform infrared spectroscopy. The results showed that the optimum preparation process of glucoamylase@ZIF-8 was 1.65 mol 2-methylimidazole, 5.85 mL glucoamylase, 33°C stirring temperature, 90 min stirring time, and 84.0230% ± 0.6006% embedding rate. At 100°C, the free glucoamylase completely lost its activity, whereas the glucoamylase@ZIF-8 still had a retained enzyme activity of 12.0123% ± 0.86158%; at pH 3-6, the highest activity of glucoamylase@ZIF-8 was 95.9531% ± 0.96181%, and about 80% of glucoamylase activity could be retained under alkaline conditions. When the ethanol concentration was 13%, the retained enzyme activity was 7.9316% ± 0.19805%, significantly higher than free enzymes. The Km of glucoamylase@ZIF-8 and free enzyme were 1235.6825 and 80.317 mg/mL, respectively. Vmax was 0.2453 and 0.149 mg/(mL min), respectively. The appearance, crystal strength, and thermal stability of glucoamylase@ZIF-8 were improved after optimization, and they had high reusability.

Estimation of nutrient (N and P) fluxes into Newark Bay, USA.

This study was conducted in northern New Jersey, USA, to estimate the nutrient fluxes from the Passaic River, the Hackensack River and other sources into Newark Bay and the nutrient residence time in Newark Bay. Bi-weekly total inorganic nitrogen (TIN) and orthophosphate concentration data in the Passaic River, the Hackensack River, and Newark Bay for over 15 years (2004-2019) were collected along with daily river discharge data from the public database. The annual TIN and orthophosphate (ortho-P) loading from the Passaic River ranged from 915 × 103 kg y-1 to 251 × 104 kg y-1 and 94 × 103 kg y-1to 372 × 103 kg y-1, respectively. The annual TIN and ortho-P loading from the Hackensack River ranged from 3.13 × 103 kg y-1 to 234 × 103 kg y-1 and 0.28 × 103 kg y-1 to 6.97 × 103 kg y-1, respectively. Seasonal variation results indicated that hurricane events highly increased TIN and ortho-P loading from riverine input and reduced residence time in Newark Bay.

Generation of functional human T cell development in NOD/SCID/IL2rγnull humanized mice without using fetal tissue: Application as a model of HIV infection and persistence.

Humanization of mice with functional T cells currently relies on co-implantation of hematopoietic stem cells from fetal liver and autologous fetal thymic tissue (so-called BLT mouse model). Here, we show that NOD/SCID/IL2rγnull mice humanized with cord blood- derived CD34+ cells and implanted with allogeneic pediatric thymic tissues excised during cardiac surgeries (CCST) represent an alternative to BLT mice. CCST mice displayed a strong immune reconstitution, with functional T cells originating from CD34+ progenitor cells. They were equally susceptible to mucosal or intraperitoneal HIV infection and had significantly higher HIV-specific T cell responses. Antiretroviral therapy (ART) robustly suppressed viremia and reduced the frequencies of cells carrying integrated HIV DNA. As in BLT mice, we observed a complete viral rebound following ART interruption, suggesting the presence of HIV reservoirs. In conclusion, CCST mice represent a practical alternative to BLT mice, broadening the use of humanized mice for research.

Convolutional neural network-based segmentation network applied to image recognition of angiodysplasias lesion under capsule endoscopy.

BACKGROUND: Small intestinal vascular malformations (angiodysplasias) are common causes of small intestinal bleeding. While capsule endoscopy has become the primary diagnostic method for angiodysplasia, manual reading of the entire gastrointestinal tract is time-consuming and requires a heavy workload, which affects the accuracy of diagnosis. AIM: To evaluate whether artificial intelligence can assist the diagnosis and increase the detection rate of angiodysplasias in the small intestine, achieve automatic disease detection, and shorten the capsule endoscopy (CE) reading time. METHODS: A convolutional neural network semantic segmentation model with a feature fusion method, which automatically recognizes the category of vascular dysplasia under CE and draws the lesion contour, thus improving the efficiency and accuracy of identifying small intestinal vascular malformation lesions, was proposed. Resnet-50 was used as the skeleton network to design the fusion mechanism, fuse the shallow and depth features, and classify the images at the pixel level to achieve the segmentation and recognition of vascular dysplasia. The training set and test set were constructed and compared with PSPNet, Deeplab3+, and UperNet. RESULTS: The test set constructed in the study achieved satisfactory results, where pixel accuracy was 99%, mean intersection over union was 0.69, negative predictive value was 98.74%, and positive predictive value was 94.27%. The model parameter was 46.38 M, the float calculation was 467.2 G, and the time length to segment and recognize a picture was 0.6 s. CONCLUSION: Constructing a segmentation network based on deep learning to segment and recognize angiodysplasias lesions is an effective and feasible method for diagnosing angiodysplasias lesions.

Preparation and characterization of glucoamylase microcapsules prepared by W/O/W type complex coacervation freeze drying.

Glucoamylase was often used in the brewing industry but was unstable to several environmental factors and reacted quickly to produce fermentable sugar, which limited its applications. Microencapsulation could effectively overcome the drawbacks. This study evaluated the feasibility of the preparation of glucoamylase microcapsules (GM) using W/O/W complex coacervation-freeze-drying method. The parameters of the microcapsules were optimized by the response surface optimization design: core-wall ratio at 1:1, wall-material concentration at 8%, and coagulation time for 20 min. Under current condition, the final microencapsulation efficiency reached 85.64 ± 1.60%. Glucoamylase could be slowly released for more than 96 h in the liquid state, and could react slowly for more than 336 h in the solid state. The optimized GM were incubated for 1 h, and the relative enzyme activity was better than that of free glucoamylase under high temperature conditions. The water capacity, solubility, morphology, differential scanning calorimetry, and Fourier transform infrared spectroscopy were conducted. Glucoamylase exhibited good sustained release characteristics. The microcapsules were more resistant to environmental stimuli and showed stronger robustness after optimization. PRACTICAL APPLICATION: Saccharification enzymes are often used in the winemaking industry, and direct addition will cause the fermentation process to heat up too quickly, resulting in the inactivation of microorganisms and saccharification enzymes, affecting the efficiency of saccharification enzymes. Therefore, microcapsules are used to encapsulate the saccharification enzyme, and its efficacy is slowly released for a long time during the fermentation process.

Chemosensory Characteristics of Brandies from Chinese Core Production Area and First Insights into Their Differences from Cognac.

This work aimed to compare the aroma characteristics of representative brandies with different grades from Yantai (one of the Chinese core production areas) and Cognac and to establish relationships between sensory descriptors and chemical composition. Descriptive analysis was performed with a trained panel to obtain the sensory profiles. Forty-three aroma-active compounds were quantified by four different methodologies. A prediction model on the basis of partial least squares analysis was performed to identify candidate compounds that were unique to a certain group of brandies. The result showed that brandies from Yantai could be distinguished from Cognac brandies on the basis of spicy, dried fruit, floral, and fruity-like aromas, which were associated with an aromatic balance between concentrations of a set of compounds such as 5-methylfurfural, γ-nonalactone, and γ-dodecalactone. Meanwhile, brandy with different grades could be distinguished on the basis of compounds derived mostly during the aging process.

Autologous humanized mouse models of iPSC-derived tumors enable characterization and modulation of cancer-immune cell interactions.

Modeling the tumor-immune cell interactions in humanized mice is complex and limits drug development. Here, we generated easily accessible tumor models by transforming either primary skin fibroblasts or induced pluripotent stem cell-derived cell lines injected in immune-deficient mice reconstituted with human autologous immune cells. Our results showed that fibroblastic, hepatic, or neural tumors were all efficiently infiltrated and partially or totally rejected by autologous immune cells in humanized mice. Characterization of tumor-immune infiltrates revealed high expression levels of the dysfunction markers Tim3 and PD-1 in T cells and an enrichment in regulatory T cells, suggesting rapid establishment of immunomodulatory phenotypes. Inhibition of PD-1 by Nivolumab in humanized mice resulted in increased immune cell infiltration and a slight decrease in tumor growth. We expect that these versatile and accessible cancer models will facilitate preclinical studies and the evaluation of autologous cancer immunotherapies across a range of different tumor cell types.

Assessing the contribution of odor-active compounds in icewine considering odor mixture-induced interactions through gas chromatography-olfactometry and Olfactoscan.

The sensory impact of odor-active compounds on icewine aroma could be influenced by perceptual interactions with other odor-active compounds. The aim of this study was to establish an approach to evaluate the contribution of odor-active compounds found in icewine considering mixture-induced perceptual interactions. By comparing the impact of key odorants detected in icewine following a gas chromatography-olfactometry approach with an Olfactoscan-based methodology using a background odor of icewine, 69 odor zones were detected, and their related compounds were further identified. The results revealed that icewine background odor could exert odor masking or enhancement on key odorants when they are considered in the complex wine aroma buffer. Several compounds can induce qualitative changes in the overall wine aroma. This study underlined the efficiency of Olfactoscan-like approaches to screen for the real impact of key odorants and to pinpoint specific compounds that could be highly influential once embedded in the aroma buffer.

Sedimentary organic carbon and nutrient distributions in an endorheic lake in semiarid area of the Mongolian Plateau.

Due to the lack of outlets, inflowing pollutants are often deposited in an endorheic lake, posing potential pressure on the environment. With climate change, extreme weather is expected to be more frequent and will contribute to the release of carbon and nutrients buried in the lakebeds. However, the distribution of sedimentary organic carbon and nutrients and the mechanisms that control the distribution are not fully understood, despite their significance to environmental development in endorheic lakes being widely recognized. In this study, the mechanisms controlling the sedimentary organic carbon and nutrient distributions in endorheic lakes were examined based on the analysis of an endorheic lake in the semiarid area of the Mongolian Plateau. The field survey results indicate that the concentrations of sedimentary organic carbon (TOC) and nutrients (NH3-N and TP) on the lakebed have significant correlations and present spatial heterogeneities. To further study the distribution mechanisms, numerical models were established to calculate the age of the water discharged from the rivers around the lake, and satellite remote sensing data were applied to examine the external source of organic carbon and nutrients and the factors influencing their movements to the lake. Based on the distribution of the water age, the water flow and mass transport trends in Lake Hulun were determined, and the time scales of the environmental processes were compared with those of water circulation. Further analysis indicates that the water circulation in the lake favors the accumulation of sedimentary organic carbon and nutrients in the northwestern part of the lake, and the organic carbon produced in the lake is transported to this region within an ice-free period. Satellite remote sensing data indicate that the region on the northwest bank of the lake experiences a larger terrestrial slope and better vegetation coverage than that on the southeast bank, which corresponds to a higher concentration of sedimentary organic carbon and nutrients in the northwest of the lake. This suggests that the sediment quality is closely related to the environment around the endorheic lake, and the larger slope and better vegetation coverage are significant factors for the high concentration of sedimentary organic carbon and nutrients on the lakebed under the conditions of scarce precipitation and low temperature. This study provides a theoretical basis and direction for further protection and management of the ecological environment of endorheic lakes.

Trained immunity induced by in vivo peptide-based STAT6 inhibition prevents ragweed allergy in mice.

BACKGROUND: Trained immunity is the ability of the innate immune system to form immune memory responses to provide support the formation of appropriate adaptive responses. Allergic airways disease (AAD) is a maladapted immune response to allergens, initiated and maintained by the type 2 (T2) inflammatory pathway. It is predicated by the elaboration of cytokines IL-4 and IL-13 and follows activation of the STAT6 transcription factor. OBJECTIVE: To investigate the role of trained immunity in mucosal immune responses following neonatal vaccination with the STAT6 inhibitory peptide (STAT6-IP), in preventing the development of ragweed-induced AAD. METHODS: We demonstrate that transfer of CD4+ T cells or dendritic cells (DC) from STAT6-IP vaccinated wild-type BALB/c mice to naïve mice, that were subsequently chronically exposed to sensitizing doses of ragweed allergen, is sufficient to prevent development of T2 responses in recipients. RESULTS: Our results demonstrate significant reductions in; airways hyperresponsiveness (AHR); ragweed-specific IgE; pulmonary inflammation; T2 cytokines; and inflammatory gene expressions in recipient mice. Expression of IDO, TGFß and T regulatory cells were all significantly increased. Anti-TGFß treatment during the ragweed sensitization phase re-constituted the pro-inflammatory T2 immune response. We show that tolerance can be attained via DC trained in the STAT6-IP-mediated tolerant milieu. This effect is not restricted to a particular allergen and does not require antigen-mediated T cell activation prior to transfer. CONCLUSION: Adoptive transfer experiments suggest that STAT6-IP treatment trains dendritic and cells to mediate tolerant immunity to chronic ragweed exposure in the airways. This indicates that early transient STAT6-inhibition constitutes an effective immunomodulatory airways allergy preventative strategy.

Identification, quantitation and sensorial contribution of lactones in brandies between China and France.

Lactones are important flavor compounds in lots of foodstuffs. They also play an important role in brandy, but have not been studied at large. In this study, solid-phase extraction (SPE) and stir bar sorptive extraction (SBSE) combined with comprehensive two-dimensional gas chromatography and time-of-flight mass spectrometry (GC × GC-TOFMS) were applied to identify and quantify lactones in brandies between China and France. Totally 17 lactones were identified, four of which were detected only in SBSE. Among them, γ-valerolactone, γ-heptalactone, δ-octalactone, γ-undecanolactone and δ-dodecalactone were detected in brandy for the first time. The results of partial least squares-discriminant analysis (PLS-DA) revealed that lactones distinguished regional characteristics among different brandies. The omission test showed that four lactones (OAV > 1) had direct impact on the aroma of brandy, and other seven lactones at sub-threshold (0.1 < OAV < 1) provided peach and apricot aroma characteristics through synergistic effects.

Myogenic progenitor cells derived from human induced pluripotent stem cell are immune-tolerated in humanized mice.

It is still unclear if immune responses will compromise the large-scale utilization of human induced pluripotent stem cells (hiPSCs)-derived cell therapies. To answer this question, we used humanized mouse models generated by the adoptive transfer of peripheral blood mononuclear cells or the cotransplantation of hematopoietic stem cells and human thymic tissue. Using these mice, we evaluated the engraftment in skeletal muscle of myoblasts derived either directly from a muscle biopsy or differentiated from hiPSCs or fibroblasts. Our results showed that while allogeneic grafts were mostly rejected and highly infiltrated with human T cells, engraftment of autologous cells was tolerated. We also observed that hiPSC-derived myogenic progenitor cells (MPCs) are not targeted by autologous T cells and natural killer cells in vitro. These findings suggest that the reprogramming and differentiation procedures we used are not immunogenic and that hiPSC-derived MPCs will be tolerated in the presence of a competent human immune system.

Penisarins A and B, Sesquiterpene Coumarins Isolated from an Endophytic Penicillium sp.

Penisarins A (1) and B (2), sesquiterpene coumarins with an unusual tricyclic sesquiterpene system, were isolated from endophytic Penicillium sp. KMU18029. Their structures were elucidated on the basis of spectroscopic methods, single-crystal X-ray diffraction, and electronic circular dichroism calculations. Compound 2 showed significant cytotoxicities against two human cancer cell lines, HL-60 and SMMC-7721, with IC50 values of 3.6 ± 0.2 and 3.7 ± 0.2 µM, respectively.

Microfibril-Associated Protein 2 (MFAP2) Potentiates Invasion and Migration of Melanoma by EMT and Wnt/ß-Catenin Pathway.

BACKGROUND Growing evidence indicates an association between microfibril-associated protein 2 (MFAP2) and a number of physiological and pathological mechanisms. The potential role of MFAP2 in cancer requires further elucidation. The present study investigated the biological behavior of MFAP2 in melanoma patients. MATERIAL AND METHODS MFAP2 inhibition was established in the B16 melanoma cell line through the use of RNA interference and was assessed by quantitative real-time PCR (qRT-PCR) and Western blot analysis. Wound-healing analysis, transwell assay, and in vivo imaging were performed to investigate the roles of MFAP2 reducing cell mobility, migration, and invasion abilities in vitro and in vivo. RESULTS We found substantially higher MFAP2 expression in B16 melanoma cells. The knockdown of MFAP2 inhibited B16 melanoma cells migration and invasion. Western blot analysis was used to assess changes in biomarkers of EMT, indicating the function of MFAP2 in EMT. We found that downregulation of MFAP2 altered the expression of Wnt/ß-catenin-linked protein. CONCLUSIONS Our results suggest that MFAP2 has potential as a molecular target to treat melanoma and suppress metastasis of melanoma cells.