Self-renewing resident arterial macrophages arise from embryonic CX3CR1(+) precursors and circulating monocytes immediately after birth.

Resident macrophages densely populate the normal arterial wall, yet their origins and the mechanisms that sustain them are poorly understood. Here we use gene-expression profiling to show that arterial macrophages constitute a distinct population among macrophages. Using multiple fate-mapping approaches, we show that arterial macrophages arise embryonically from CX3CR1(+) precursors and postnatally from bone marrow-derived monocytes that colonize the tissue immediately after birth. In adulthood, proliferation (rather than monocyte recruitment) sustains arterial macrophages in the steady state and after severe depletion following sepsis. After infection, arterial macrophages return rapidly to functional homeostasis. Finally, survival of resident arterial macrophages depends on a CX3CR1-CX3CL1 axis within the vascular niche.

First report of Tulip Virus X infecting tulip (Tulipa gesneriana) in Korea.

Tulip virus X (tulip virus X, TVX) is a member of the genus Potexvirus (family Alphaflexiviridae) and is a positive single-stranded RNA virus. TVX was described first in Scotland (Mowat 1982), followed by several countries (Yamaji et al. 2001; Tzanetakis et al. 2005; Ward et al. 2008; Dees et al. 2011; Sochacki and Komorowska 2012; Wylie et al. 2019). In April 2021, 86 whole tulip plants showing viral symptoms in leaves (mosaic, yellowing, and malformation) and flowers (color breaking) were collected in Chilgok, Chuncheon, Goseong, Yecheon and Yesan, Korea. Furthermore, high-throughput sequencing was performed to identify viruses that infect tulips in Korea. Total RNA was extracted from pooled the leaves and petals using a Maxwell® 16 LEV Plant RNA Kit (Promega, Madison, USA). We constructed a single library using the TruSeq Stranded Total RNA LT Sample Prep Kit for Plant (Illumina, San Diego, USA). The library was 100 bp paired-end sequenced using Illumina's NovaSeq 6000 (Macrogen, Seoul, Korea) and was assembled de novo using Trinity software version trinityrnaseq_r20140717, with default parameters. The contigs were annotated as in previous study (Lee et al. 2020), revealing a single contig each related to TVX, lily symptomless virus (LSV), and tulip breaking virus (TBV) was generated from 648 million total reads. The TVX-related contig (GenBank ON205948) consisting of 6,076 bp showed 99.52% nucleotide identity (6027/6056 bp) with TVX-J (GenBank AB066288). We conducted an RT-PCR assay to validate the presence of viruses with specific primers as TVX-F5093/R5624 (5'-CTATCCGGACTCATTCTACTTC/GTGCGTTCCAGATAAGCTTG-3'), LSV-F7013/R7338 (5'-CTTGGTCGACAGGGACATAAC/GATTGGAATTGTGCTTTTCAGC-3'), and TBV-F7515/R8116 (5'-GTGTGTCATGGATGATTGTTG/CAACTGATTTGCTACCGCTAG-3'). Consequently, TVX were detected in 13 of 86 samples. Moreover, LSV and TBV were detected in 15 and 26 samples, respectively. However, the yellowing and mosaic observed in the TVX infected samples were not observed in the LSV and TBV infected samples. Subsequently, two TVX amplicons were selected, cloned and sequenced. The obtained sequences were 532 bp and were named YS24 and YS38 (GenBank LC664027 and LC664028), respectively. The Korean isolates showed 98.68% (525/532 bp) and 99.62% (530/532 bp) identity with Australian isolate (GenBank MH886522) in BLASTn analysis. To bioassay for TVX, the infected tulip leaf tissue from which YS24 was obtained was used to sap-inoculate, in triplicates, 15 species of indicator plants (Nicotiana benthamiana, N. clevelandii, N. debneyi, N. glutinosa, N. rustica, N. tabacum, Datura stramonium, Glycine max, Phaseolus vulgaris, Chenopodium amaranticolor, C. quinoa, Cucumis sativus, Cu. melo, Gomphrena globosa, and Tetragonia tetragonioides). After 14 days of inoculation, we observed distinct chlorotic spots on inoculated and upper leaves of C. quinoa, but no symptoms were observed in other indicator plants. In RT-PCR assay using TVX-specific primers, only C. quinoa showed a positive reaction. In previous studies, C. amaranticolor, C. quinoa, G. globosa, and N. benthamiana were known as the experimental host of TVX (Dees et al. 2011; Tzanetakis et al. 2005), but only C. quinoa was confirmed to be susceptible to the Korean isolate. Furthermore, transmission electron microscopy revealed typical flexuous rod-shaped viral particles in the inoculated C. quinoa. To our knowledge, this is the first report of TVX infecting tulips in Korea.

Downregulation of MHC Class II by Ubiquitination Is Required for the Migration of CD206+ Dendritic Cells to Skin-Draining Lymph Nodes.

Dendritic cells (DCs) are critical players in skin homeostasis. A subset of mannose receptor (CD206)-expressing monocyte-derived DCs was found in skin, and their migratory counterpart is present in skin-draining lymph nodes (sdLNs). Skin CD206+ DCs were shown to upregulate MHC class II (MHCII) progressively, raising the question of whether this feature affects their biology. In this study, we assessed the role of MHCII regulation in the development and migration of these cells in mouse models expressing differential MHCII levels. Using CD206 as a surrogate marker, we found that skin CD206+ DCs develop in an MHCII-independent manner. However, their migration to sdLNs was affected by overexpression rather than absence or lower expression of MHCII. Accordingly, B16 tumor growth was exacerbated in mice overexpressing MHCII in the absence of ubiquitination. Mechanistically, CD206+ DCs from these mice showed decreased IRF4 and CCR7 expression. LPS, which is known to promote monocyte-derived DC recruitment to sdLNs, partially improved these defects. However, GM-CSF delivery restored CD206+ DC migration by promoting IRF4 expression. Collectively, these data show that MHCII downregulation is crucial for IRF4-dependent migration of CD206+ DCs to sdLNs in health and disease.

Transcriptome Analysis Reveals Nonfoamy Rather Than Foamy Plaque Macrophages Are Proinflammatory in Atherosclerotic Murine Models.

RATIONALE: Monocyte infiltration into the subintimal space and its intracellular lipid accumulation are the most prominent features of atherosclerosis. To understand the pathophysiology of atherosclerotic disease, we need to understand the characteristics of lipid-laden foamy macrophages in the subintimal space during atherosclerosis. OBJECTIVE: We sought to examine the transcriptomic profiles of foamy and nonfoamy macrophages isolated from atherosclerotic intima. METHODS AND RESULTS: Single-cell RNA sequencing analysis of CD45+ leukocytes from murine atherosclerotic aorta revealed that there are macrophage subpopulations with distinct differentially expressed genes involved in various functional pathways. To specifically characterize the intimal foamy macrophages of plaque, we developed a lipid staining-based flow cytometric method for analyzing the lipid-laden foam cells of atherosclerotic aortas. We used the fluorescent lipid probe BODIPY493/503 and assessed side-scattered light as an indication of cellular granularity. BODIPYhiSSChi foamy macrophages were found residing in intima and expressing CD11c. Foamy macrophage accumulation determined by flow cytometry was positively correlated with the severity of atherosclerosis. Bulk RNA sequencing analysis showed that compared with nonfoamy macrophages, foamy macrophages expressed few inflammatory genes but many lipid-processing genes. Intimal nonfoamy macrophages formed the major population expressing IL (interleukin)-1ß and many other inflammatory transcripts in atherosclerotic aorta. CONCLUSIONS: RNA sequencing analysis of intimal macrophages from atherosclerotic aorta revealed that lipid-loaded plaque macrophages are not likely the plaque macrophages that drive lesional inflammation.

Conventional Dendritic Cells Impair Recovery after Myocardial Infarction.

Ischemic myocardial injury results in sterile cardiac inflammation that leads to tissue repair, two processes controlled by mononuclear phagocytes. Despite global burden of cardiovascular diseases, we do not understand the functional contribution to pathogenesis of specific cardiac mononuclear phagocyte lineages, in particular dendritic cells. To address this limitation, we used detailed lineage tracing and genetic studies to identify bona fide murine and human CD103+ conventional dendritic cell (cDC)1s, CD11b+ cDC2s, and plasmacytoid DCs (pDCs) in the heart of normal mice and immunocompromised NSG mice reconstituted with human CD34+ cells, respectively. After myocardial infarction (MI), the specific depletion of cDCs, but not pDCs, improved cardiac function and prevented adverse cardiac remodeling. Our results showed that fractional shortening measured after MI was not influenced by the absence of pDCs. Interestingly, however, depletion of cDCs significantly improved reduction in fractional shortening. Moreover, fibrosis and cell areas were reduced in infarcted zones. This correlated with reduced numbers of cardiac macrophages, neutrophils, and T cells, indicating a blunted inflammatory response. Accordingly, mRNA levels of proinflammatory cytokines IL-1ß and IFN-γ were reduced. Collectively, our results demonstrate the unequivocal pathological role of cDCs following MI.

Multiple drilling and multiple matchstick-like bone allografts for large osteonecrotic lesions in the femoral head: an average 3-year follow-up study.

INTRODUCTION: We aimed to present the clinical outcomes of multiple drilling and multiple matchstick-like bone allograft for large osteonecrotic lesions of the femoral head as a joint-preserving surgery. MATERIALS AND METHODS: Between March 2014 and March 2018, 57 patients (77 hips) who underwent multiple drilling and multiple matchstick-like bone allograft for large lesions (≥ 30%) in osteonecrosis of the femoral head (ONFH) were included. Harris hip scores (HHS) and Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC) were obtained preoperatively and at the latest follow-up. Plain radiographs were obtained every 3 months. Femoral head collapse ≥ 2 mm was defined as a radiological failure, and conversion to total hip arthroplasty (THA) was regarded as a clinical failure. RESULTS: After exclusion of 5 patients (5 hips) who lost to follow-up, 52 patients (34 men, 18 women; 72 hips) were finally enrolled. The mean follow-up period was 3.4 (range 2-4.5) years. Nineteen hips (28.4%) required conversion to THA at a mean of 21.6 (range 6-42) months postoperatively. In the remaining 53 hips (71.6%) with clinical success, the mean HHS and WOMAC improved from 63 and 31.3 preoperatively to 80.6 and 16.3 at the final follow-up, respectively (p < 0.001). Radiological failure occurred in four hips (6%). The overall failure rate was 31.9% (23/72 hips), and the mean survival duration until failure was 21.2 months (6-42 months). The lesion size, lesion location, and the use of corticosteroids as the cause of ONFH were associated with clinical failure. CONCLUSION: Multiple drilling and multiple matchstick-like bone allograft may be a useful treatment option for alleviating the symptoms in ONFH patients with large lesions who want to preserve their hips.

Adaptive Noise Reduction Algorithm to Improve R Peak Detection in ECG Measured by Capacitive ECG Sensors.

Electrocardiograms (ECGs) can be conveniently obtained using capacitive ECG sensors. However, motion noise in measured ECGs can degrade R peak detection. To reduce noise, properties of reference signal and ECG measured by the sensors are analyzed and a new method of active noise cancellation (ANC) is proposed in this study. In the proposed algorithm, the original ECG signal at QRS interval is regarded as impulsive noise because the adaptive filter updates its weight as if impulsive noise is added. As the proposed algorithm does not affect impulsive noise, the original signal is not reduced during ANC. Therefore, the proposed algorithm can conserve the power of the original signal within the QRS interval and reduce only the power of noise at other intervals. The proposed algorithm was verified through comparisons with recent research using data from both indoor and outdoor experiments. The proposed algorithm will benefit a noise reduction of noisy biomedical signal measured from sensors.

Changes in cellular microRNA expression induced by porcine circovirus type 2-encoded proteins.

Porcine circovirus type 2 (PCV2) is the primary causative agent of postweaning multisystemic wasting syndrome, which leads to serious economic losses in the pig industry worldwide. While the molecular basis of PCV2 replication and pathogenicity remains elusive, it is increasingly apparent that the microRNA (miRNA) pathway plays a key role in controlling virus-host interactions, in addition to a wide range of cellular processes. Here, we employed Solexa deep sequencing technology to determine which cellular miRNAs were differentially regulated after expression of each of three PCV2-encoded open reading frames (ORFs) in porcine kidney epithelial (PK15) cells. We identified 51 ORF1-regulated miRNAs, 74 ORF2-regulated miRNAs, and 32 ORF3-regulated miRNAs that differed in abundance compared to the control. Gene ontology analysis of the putative targets of these miRNAs identified transcriptional regulation as the most significantly enriched biological process, while KEGG pathway analysis revealed significant enrichment for several pathways including MAPK signaling, which is activated during PCV2 infection. Among the potential target genes of ORF-regulated miRNAs, two genes encoding proteins that are known to interact with PCV2-encoded proteins, zinc finger protein 265 (ZNF265) and regulator of G protein signaling 16 (RGS16), were selected for further analysis. We provide evidence that ZNF265 and RGS16 are direct targets of miR-139-5p and let-7e, respectively, which are both down-regulated by ORF2. Our data will initiate further studies to elucidate the roles of ORF-regulated cellular miRNAs in PCV2-host interactions.

Ribonanza: deep learning of RNA structure through dual crowdsourcing.

Prediction of RNA structure from sequence remains an unsolved problem, and progress has been slowed by a paucity of experimental data. Here, we present Ribonanza, a dataset of chemical mapping measurements on two million diverse RNA sequences collected through Eterna and other crowdsourced initiatives. Ribonanza measurements enabled solicitation, training, and prospective evaluation of diverse deep neural networks through a Kaggle challenge, followed by distillation into a single, self-contained model called RibonanzaNet. When fine tuned on auxiliary datasets, RibonanzaNet achieves state-of-the-art performance in modeling experimental sequence dropout, RNA hydrolytic degradation, and RNA secondary structure, with implications for modeling RNA tertiary structure.

The luminescent characterization of (Sr, Ca)2SiO4:Eu2+ nanopowders synthesized by a co-precipitation method.

(Sr, Ca)2SiO4:Eu2+ nanopowders were prepared by a co-precipitation method, and then the effects of Ca2+ ions on the structural and luminescent properties were investigated. The pure Sr2SiO4:Eu2+ powders were perfectly composed of the beta-phase, whereas the substitution of Ca2+ ions led to the beta --> alpha' phase transition. The photoluminescence spectra of Sr2SiO4:Eu2+ exhibited two excitation bands at around 330 and 375 nm assigned to Eu(I) and (II) sites, respectively, resulting in two emission bands at around 473 and 543 nm. Meanwhile, the dominant peak wavelengths of the emission spectra of (Sr, Ca)2SiO4:Eu2+ could be tuned, depending on the cation ratio of Ca2+ to Sr2+. The substitution of Ca2+ ions for Sr2+ ions caused the red-shift of the emission peaks of Sr(2-x)Ca(x)SiO4:Eu2+ powders with increasing Ca2+ content (x = 0-1.0) due to the increase in the crystal field strength.

Effects of polymorphisms in the porcine microRNA MIR206 / MIR133B cluster on muscle fiber and meat quality traits.

MicroRNAs are a class of small non-coding RNA molecules that repress gene expression primarily at the post-transcriptional level. Genetic variations in microRNA genes may contribute to phenotypic differences by altering the expression of microRNAs and their targets. Here, we identified 12 single nucleotide polymorphisms (SNPs) in the genomic region of the porcine MIR206 / MIR133B cluster, 10 and 2 of which were associated with MIR206 and MIR133B respectively. All 12 SNPs were located within primary microRNAs. Allele frequency determination in different pig breeds (Berkshire, n = 153; Landrace, n = 125; Yorkshire, n = 173) and association studies of muscle fiber characteristics, lean meat production and meat quality traits were performed on the MIR206 and MIR133B SNPs. The MIR206 SNPs were associated with the percentage of type IIa and IIb fibers for muscle fiber area composition, meat quality traits including drip loss and lightness, and backfat thickness, a parameter of lean meat production. In addition, we found significant association of the MIR133B SNPs with total muscle fiber number, loin eye area, and muscle pH. Furthermore, these SNPs significantly affected the levels of mature MIR206 and MIR133B , respectively, primarily by regulating the processing of primary microRNAs into precursor microRNAs. Interestingly, altered MIR206 levels correlated with phenotypic variability among genotypes of the MIR206 SNP. Our data suggest that polymorphisms in the porcine MIR206 / MIR133B cluster are a genetic factor affecting muscle and meat quality traits.

Effects of polymorphisms in the 3' untranslated region of the porcine PPARGC1A gene on muscle fiber characteristics and meat quality traits.

Peroxisome proliferator-activated receptor γ coactivator 1 α (PPARGC1A) is a transcriptional coactivator that is involved in a variety of biological processes including muscle fiber type composition. Here, we identified two single nucleotide polymorphisms (SNPs; *2690T>C and *2864T>C) and one insertion/deletion in the 3' untranslated region of porcine PPARGC1A. These SNPs were genotyped by direct sequencing in a total of 439 pigs representing three different pig breeds (Berkshire, n = 156; Yorkshire, n = 163; Landrace, n = 120). We evaluated the effects of diplotypes of individual PPARGC1A 3'UTR SNPs on muscle fiber characteristics and meat quality traits. The *2690T>C polymorphism was significantly associated with the percentage of type I and IIb fibers for both muscle fiber number and area composition (P < 0.05), and also showed a significant association with muscle pH, a parameter of meat quality (P = 0.0188). The *2864T>C polymorphism was also associated with meat quality traits including muscle pH (P = 0.0071), drip loss (P = 0.0006), and lightness (P = 0.0702), but showed no significant association with muscle fiber characteristics. Interestingly, each SNP affected PPARGC1A expression significantly at the protein level but not at the mRNA level, thereby accounting for phenotypic variability among genotypes. Taken together, our data suggest that the *2690T>C and *2864T>C polymorphisms can be used as genetic markers for selection toward improved meat quality.

Synthesis and luminescent characterization of Sr2SiO4:Eu2+ nanopowders.

Sr2SiO4:Eu2+ nanopowders were synthesized by a co-precipitation method using strontium nitrate, 3-aminopropyltriethoxysilane (APTES), europium nitrate hydrate, and a flux (NH4Cl). The structural and luminescent properties strongly depended on the firing conditions, the amount of APTES, and the flux content. Alpha'-Sr2SiO4 was produced as a dominant phase after firing as-prepared powders without a flux, whereas the addition of NH4Cl caused beta-Sr2SiO4 to be the primary phase. At a small amount of APTES, the as-prepared powders that were fired with NH4Cl consisted of the beta-Sr2SiO4, Sr3SiO5, and un-reacted SrO phases. Then, the phase transformation from SrO and Sr3SiO5 to beta-Sr2SiO4 gradually proceeded as the amount of APTES was increased, leading to the pure beta-Sr2SiO4 phase at 0.5 M APTES. The phase transition and photoluminescence properties strongly relied on the amount of NH4Cl. The preparation condition of 0.5 M APTES and 2 wt% NH4Cl was the optimum condition to obtain the pure beta-Sr2SiO4:Eu2+ phase and the most excellent PL intensity.

Dependence of the up-conversion properties of NaYF4:Yb3+/Er3+ nanopowders on the structure and particle size.

NaYF4:Yb3+/Er3+ powders were synthesized by a solvothermal method. The effects of an amount of NaF, solvothermal temperatures, and solvothermal time on the up-conversion emission and the phase transition were investigated. Hexagonal rods (beta-form) with well-defined facets were obtained at the solvothermal condition of 210 degrees C and 24 h, whereas cubic nanocrystals (alpha-form) were produced at the low solvothermal temperature and short solvothermal time. These results demonstrated that the high solvothermal temperature of 210 degrees C supplied the enough thermodynamic energy to activate the alpha --> beta transformation, while long solvothermal time was necessary to complete the kinetic process for this phase transition. An amount of NaF obviously contributed to the phase transition, particle size, and the particle morphology. The beta-form with irregular shapes was superior to the well-defined beta-form in the emission intensity due to the enhanced surface scattering. This work suggested that the beta-form could not be obtained through its own nucleation and growth in the solutions during the solvothermal treatment, but could be produced through the alpha --> beta phase transition.

Machine Learning-Based Automatic Rating for Cardinal Symptoms of Parkinson Disease.

OBJECTIVE: We developed and investigated the feasibility of a machine learning-based automated rating for the 2 cardinal symptoms of Parkinson disease (PD): resting tremor and bradykinesia. METHODS: Using OpenPose, a deep learning-based human pose estimation program, we analyzed video clips for resting tremor and finger tapping of the bilateral upper limbs of 55 patients with PD (110 arms). Key motion parameters, including resting tremor amplitude and finger tapping speed, amplitude, and fatigue, were extracted to develop a machine learning-based automatic Unified Parkinson's Disease Rating Scale (UPDRS) rating using support vector machine (SVM) method. To evaluate the performance of this model, we calculated weighted κ and intraclass correlation coefficients (ICCs) between the model and the gold standard rating by a movement disorder specialist who is trained and certified by the Movement Disorder Society for UPDRS rating. These values were compared to weighted κ and ICC between a nontrained human rater and the gold standard rating. RESULTS: For resting tremors, the SVM model showed a very good to excellent reliability range with the gold standard rating (κ 0.791; ICC 0.927), with both values higher than that of nontrained human rater (κ 0.662; ICC 0.861). For finger tapping, the SVM model showed a very good reliability range with the gold standard rating (κ 0.700 and ICC 0.793), which was comparable to that for nontrained human raters (κ 0.627; ICC 0.797). CONCLUSION: Machine learning-based algorithms that automatically rate PD cardinal symptoms are feasible, with more accurate results than nontrained human ratings. CLASSIFICATION OF EVIDENCE: This study provides Class II evidence that machine learning-based automated rating of resting tremor and bradykinesia in people with PD has very good reliability compared to a rating by a movement disorder specialist.

Identification of a stroma-mediated Wnt/beta-catenin signal promoting self-renewal of hematopoietic stem cells in the stem cell niche.

With contrasting observations on the effects of beta-catenin on hematopoietic stem cells (HSCs), the precise role of Wnt/beta-catenin signals on HSC regulation remains unclear. Here, we show a distinct mode of Wnt/beta-catenin signal that can regulate HSCs in a stroma-dependent manner. Stabilization of beta-catenin in the bone marrow stromal cells promoted maintenance and self-renewal of HSCs in a contact-dependent manner, whereas direct stabilization in hematopoietic cells caused loss of HSCs. Interestingly, canonical Wnt receptors and beta-catenin accumulation were predominantly enriched in the stromal rather than the hematopoietic compartment of bone marrows. Moreover, the active form of beta-catenin accumulated selectively in the trabecular endosteum in "Wnt 3a-stimulated" or "irradiation-stressed," but not in "steady-state" marrows. Notably, notch ligands were induced in Wnt/beta-catenin activated bone marrow stroma and downstream notch signal activation was seen in the HSCs in contact with the activated stroma. Taken together, Wnt/beta-catenin activated stroma and their cross-talk with HSCs may function as a physiologically regulated microenvironmental cue for HSC self-renewal in the stem cell niche.

Fuzzy support vector machine-based personalizing method to address the inter-subject variance problem of physiological signals in a driver monitoring system.

Physiological signals can be utilized to monitor conditions of a driver, but the inter-subject variance of physiological signals can degrade the classification accuracy of the monitoring system. Personalization of the system using the data of a tested subject, called local data, can be a solution, but the acquisition of sufficient local data may not be possible in real situations. Therefore, this paper proposes an effective personalizing method using small-sized local data. The proposed method utilizes a fuzzy support vector machine to allocate higher weight to the local data than to others, and a fuzzy membership is assigned to the training data by analyzing the importance of each datum. Three classification problems for a physiological signal-based driver monitoring system are introduced and utilized to validate the proposed method. The classification accuracy is compared with that of other personalizing methods, and the results show that the proposed method achieves a better accuracy on average, which is 3.46% higher than that of the simple approach using a basic support vector machine, thereby proving its effectiveness. The proposed method can train a personalized classifier with improved accuracy for a tested subject. The advantages of the proposed method can be utilized to develop a practical driver monitoring system.

Lack of the E3 Ubiquitin Ligase March1 Affects CD8 T Cell Fate and Exacerbates Insulin Resistance in Obese Mice.

Obesity is a major risk factor for the development of insulin resistance and type 2 diabetes. However, the mechanisms that trigger the underlying adipose tissues inflammation are not completely understood. Here, we show that the E3 ubiquitin ligase March1 controls the phenotypic and functional properties of CD8+ T cells in mice white adipose tissue. In a diet-induced obesity model, mice lacking March1 [March1 knockout (KO)] show increased insulin resistance compared to their WT counterparts. Also, in obese March1 KO mice, the proportions of effector/memory (Tem) and resident/memory (Trm) CD8+ T cells were higher in the visceral adipose tissue, but not in the spleen. The effect of March1 on insulin resistance and on the phenotype of adipose tissue CD8+ T cells was independent of major histocompatibility complex class II ubiquitination. Interestingly, we adoptively transferred either WT or March1 KO splenic CD8+ T cells into obese WT chimeras that had been reconstituted with Rag1-deficient bone marrow. We observed an enrichment of Tem and Trm cells and exacerbated insulin resistance in mice that received March1 KO CD8 T cells. Mechanistically, we found that March1 deficiency alters the metabolic activity of CD8+ T cells. Our results provide additional evidence of the involvement of CD8+ T cells in adipose tissue inflammation and suggest that March1 controls the metabolic reprogramming of these cells.

Isolation and Characterization of Aortic Dendritic Cells and Lymphocytes in Atherosclerosis.

Dendritic cells (DCs) are central to initiate antigen-specific immunity and tolerance. The in vivo development and distribution of DCs are now better understood even in nonlymphoid tissues [1]. Atherosclerosis is a chronic inflammatory disease of blood vessels and DCs are highly enriched in the intimal area of the aorta, which is predisposed to develop atherosclerosis. Previously, we were the first to show antigen presenting DCs and their subsets in the aorta [2, 3]. Here, we discuss several useful methods to characterize not only DCs but also other immune cells in steady state and atherosclerotic aorta. These comprise multiparameter flow cytometry strategies including intracellular staining and cell sorting, en face immunohistochemistry of DCs and regulatory T cells (Tregs), and Oil Red O staining of atherosclerotic lesions in the aorta.

From the Cover: Lifelong Exposure of C57bl/6n Male Mice to Bisphenol A or Bisphenol S Reduces Recovery From a Myocardial Infarction.

Bisphenol A (BPA) leaches from plastics to contaminate foodstuffs. Analogs, such as bisphenol S (BPS), are now used increasingly in manufacturing. Greater BPA exposure has been correlated with exacerbation of cardiovascular disease, including myocardial infarction (MI). To test the hypothesis that bisphenol exposure impairs cardiac healing, we exposed C57bl/6n mice to water containing 25ng/ml BPA or BPS from conception and surgically induced an MI in adult male progeny. Increased early death and cardiac dilation, and reduced cardiac function were found post-MI in BPA- and BPS-exposed mice. Flow cytometry revealed increased monocyte and macrophage infiltration that correlated with increased chemokine C-C motif ligand-2 expression in the infarct. In vitro BPA and BPS addition increased matrix metalloproteinase-9 (MMP) protein and secreted activity in RAW264.7 macrophage cells suggesting that invivo increases in MMP2 and MMP9 in exposed infarcts were myeloid-derived. Bone marrow-derived monocytes isolated from exposed mice had greater expression of pro-inflammatory polarization markers when chemokine stimulated indicating an enhanced susceptibility to develop a pro-inflammatory monocyte population. Chronic BPA exposure of estrogen receptor beta (ERß) deficient mice did not worsen early death, cardiac structure/function, or expression of myeloid markers after an MI. In contrast, BPS exposure of ERß-deficient mice resulted in greater death and expression of myeloid markers. We conclude that lifelong exposure to BPA or BPS augmented the monocyte/macrophage inflammatory response and adverse remodeling from an MI thereby reducing the ability to survive and successfully recover, and that the adverse effect of BPA, but not BPS, is downstream of ERß signaling.