Stable Phenotype Research Articles

Unusual, Stable Phenotype in Patient with Ser70Arg TTR Mutation on Diflunisal Therapy (4490)

Unusual, Stable Phenotype in Patient with Ser70Arg TTR Mutation on Diflunisal Therapy (4490)

Left Ventricular Remodeling in Hypertrophic Cardiomyopathy: An Overview of Current Knowledge

While most patients with hypertrophic cardiomyopathy (HCM) show a relatively stable morphologic and clinical phenotype, in some others, progressive changes in the left ventricular (LV) wall thickness, cavity size, and function, defined, overall, as “LV remodeling”, may occur. The interplay of multiple pathophysiologic mechanisms, from genetic background to myocardial ischemia and fibrosis, is implicated in this process. Different patterns of LV remodeling have been recognized and are associated with a specific impact on the clinical course and management of the disease. These findings underline the need for and the importance of serial multimodal clinical and instrumental evaluations to identify and further characterize the LV remodeling phenomenon. A more complete definition of the stages of the disease may present a chance to improve the management of HCM patients.

Melanoma Differentiation Trajectories Determine Sensitivity toward Pre-Existing CD8+ Tumor-Infiltrating Lymphocytes

The highly plastic nature of melanoma enables its transition among diverse cell states to survive hostile conditions. However, the interplay between specific tumor cell states and intratumoral T cells remains poorly defined. With MAPK inhibitor‒treated BRAFV600-mutant tumors as models, we linked human melanoma state transition to CD8+ T cell responses. Repeatedly, we observed that isogenic melanoma cells could evolve along distinct differentiation trajectories on single BRAF inhibitor (BRAFi) treatment or dual BRAFi/MEKi treatment, resulting in BRAFi‒induced hyperdifferentiated and BRAFi/MEKi‒induced dedifferentiated resistant subtypes. Taking advantage of patient-derived autologous CD8+ tumor-infiltrating lymphocytes (TILs), we demonstrate that progressive melanoma cell state transition profoundly affects TIL function. Tumor cells along the hyperdifferentiation trajectory continuously gained sensitivity toward tumor-reactive CD8+ TILs, whereas those in the dedifferentiation trajectory acquired T cell resistance in part owing to the loss of differentiation antigens. Overall, our data reveal the tight connection of MAPKi‒induced temporary (drug-tolerant transition state) and stable (resistant state) phenotype alterations with T cell function and further broaden the current knowledge on melanoma plasticity in terms of sculpting local antitumor immune responses, with implications for guiding the optimal combination of targeted therapy and immunotherapy.

Mutation status and prognostic value of KRAS and NRAS mutations in Moroccan colon cancer patients: A first report.

This study aimed to estimate the incidence of KRAS, NRAS, and BRAF mutations in the Moroccan population, and investigate the associations of KRAS and NRAS gene mutations with clinicopathological characteristics and their prognosis value. To achieve these objectives, we reviewed medical and pathology reports for 210 patients. RAS testing was investigated by Sanger sequencing and Pyrosequencing technology. BRAF (exon 15) status was analyzed by the Sanger method. The expression of MMR proteins was evaluated by Immunohistochemistry. KRAS and NRAS mutations were found in 36.7% and 2.9% of 210 patients, respectively. KRAS exon 2 mutations were identified in 76.5% of the cases. RAS-mutated colon cancers were significantly associated with female gender, presence of vascular invasion, classical adenocarcinoma, moderately differentiated tumors, advanced TNM stage III-IV, left colon site, higher incidence of distant metastases at the time of diagnostic, microsatellite stable phenotype, lower number of total lymph nodes, and higher means of positive lymph nodes and lymph node ratio. KRAS exon 2-mutated colon cancers, compared with KRAS wild-type colon cancers were associated with the same clinicopathological features of RAS-mutated colon cancers. NRAS-mutated patients were associated with lower total lymph node rate and the presence of positive lymph node. Rare RAS-mutated tumors, compared with wild-type tumors were more frequently moderately differentiated and associated with lower lymph node rate. We found that KRAS codon 13-mutated, tumors compared to codon 12-mutated tumors were significantly correlated with a higher death cases number, a lower rate of positive lymph, lower follow-up time, and poor overall survival. Our findings show that KRAS and NRAS mutations have distinct clinicopathological features. KRAS codon 13-mutated status was the worst predictor of prognosis at all stages in our population.

Draft Genome Sequences of Blastococcus sp. Clones TML/M2B and TML/C7B, with Different Motilities, Isolated in a Laboratory

Two novel Blastococcus sp. clones, TML/M2B and TML/C7B, with 2 stable different growth phenotypes, were isolated from a laboratory tissue culture. The draft genome sequences generated through genomic sequencing of clones TML/M2B and TML/C7B contain 4 and 2 contigs, respectively. The respective genome sizes are 4.10 Mb and 4.11 Mb, with GC contents of 74.17% and 74.14%.

High-Throughput Culture Method of Induced Pluripotent Stem Cell Derived Alveolar Epithelial Cells Using Floating Matrigel Droplets

<h3>Purpose</h3> Induced pluripotent stem cell derived alveolar epithelial cells (iPSC-AECs) are a patient specific cell source for bio-engineering of human pulmonary epithelium. Disease modeling and therapeutic applications require cost effective and technically feasible differentiation and expansion protocols. Current culture protocols are labor and resource intensive and not scalable for large animal or human studies. We developed a scalable culture protocol switching from adherent hydrogel drops to suspension culture. <h3>Methods</h3> Genetically modified iPSCs with fluorescent reporters for Nkx2.1 and SPC underwent differentiation in a stepwise culture protocol that mimics lung development. Cells were purified, sorted and embedded in a liquid Matrigel precursor. Homogenous liquid precursor was added to culture plates to form adherent droplets or to a silicone mold to form cell laden Matrigel spheroids which were subsequently transferred to spinner flasks with media as floating droplets. After culture, monolayer spheres of iPSC-AECs were digested and trypsinized to form single cell suspensions. Equal numbers of iPSC-AECs from the two culture conditions were seeded into decellularized lung scaffolds. <h3>Results</h3> iPSC-AECs cultured in floating droplets were more proliferative than adherent droplets, with significantly higher total cell counts and Ki67 expression. Equivalent expression of the distal lung markers Nkx2.1, SPC, and AQP5 was observed for both culture conditions in isolated iPSC-AECs and iPSC-AECs from recellularized lung scaffolds. Similar histologic appearance was appreciated on lungs recellularized from both culture groups. The floating droplet method was more time and cost efficient. <h3>Conclusion</h3> We report a simple, scalable, new culture method for iPSC-AECs with stable cellular phenotype that could support large animal and human studies. Our protocol enhanced proliferation and reduced labor requirements compared to previously reported methods.

Slicing and dicing myositis for cures and prevention.

Myositis is a group of conditions that vary greatly in risk factors, clinical manifestations, laboratory markers, presumed pathogenetic mechanisms, treatment responses and prognoses. Approaches to divide myositis into mutually exclusive and stable phenotypes are being considered, but are we thinking comprehensively enough in our attempts at classification?

Retraction: Individual personalities shape task differentiation in a social spider.

You have accessMoreSectionsView PDF ToolsAdd to favoritesDownload CitationsTrack Citations ShareShare onFacebookTwitterLinked InRedditEmail Cite this article Proceedings B Editor-in-Chief 2021Retraction: Individual personalities shape task differentiation in a social spiderProc. R. Soc. B.2882021033520210335http://doi.org/10.1098/rspb.2021.0335SectionYou have accessRetractionsRetraction: Individual personalities shape task differentiation in a social spider Proceedings B Editor-in-Chief Google Scholar Find this author on PubMed Search for more papers by this author Proceedings B Editor-in-Chief Google Scholar Find this author on PubMed Search for more papers by this author Published:10 March 2021https://doi.org/10.1098/rspb.2021.0335This article retracts the followingResearch ArticleIndividual personalities shape task differentiation in a social spiderhttps://doi.org/10.1098/rspb.2013.1407 Lena Grinsted, Jonathan N. Pruitt, Virginia Settepani and Trine Bilde volume 280issue 1767Proceedings of the Royal Society B: Biological Sciences22 September 2013Proc. R. Soc. B280, 20131407. (Published Online 22 September 2013). (doi:10.1098/rspb.2013.1407)The Editor-in-Chief and the Royal Society are retracting the article ‘Individual personalities shape task differentiation in a social spider’ by Lena Grinsted, Jonathan N. Pruitt, Virginia Settepani and Trine Bild [1].Two of the authors, T Bilde and L Grinsted, have drawn the journal's attention to an overlap between the data on boldness in this paper and in the supplementary material of a paper published in Animal Behaviour by Pruitt, JN, Grinsted L and Settepani V [2] (now retracted [3]). Very high levels of duplication between datasets were confirmed upon inspection of the data as downloaded from Dryad [4] or supplied, separately, by JN Pruitt and L Grinsted (the data are identical). 74% of the unique values in the Pruitt et al. [2] dataset also appear in the Grinsted et al. [1] dataset. On this basis, co-authors L Grinsted, V Settepani and T Bilde requested a retraction of the paper.Subsequent investigation by the editors has determined that the nature of the boldness data collected in the Grinsted et al. [1] study is very different from the impression created by reading the paper and inspecting the associated data in Dryad. The initial impression is that spiders were tested individually, with high precision (to the nearest hundredth of a second) and repeatability (presented in the Pruitt et al. [2] supplementary material). In fact, spiders were tested in groups of up to 20 at low precision, with concomitant recording of identical values of latency (boldness) for individuals moving at similar times (JN Pruitt, personal communication). Moreover, spiders used to generate the two data sets (Grinsted et al. [1], Pruitt et al. [2]) were intermingled in these test groups. This design would allow for some level of duplication within and between the data sets. We carried out simulations to estimate the timing accuracy necessary to account for the very high frequency of duplicate values. These simulations suggest that spiders must have been scored, on average, as having the same time if they moved within 15 to 20 s of each other. Therefore, the boldness data are not independent, but may reflect aspects of their testing group, and we cannot rely on the estimates of repeatability.Without the support of the repeatability analysis in Pruitt et al. [2], we cannot determine whether the single boldness value used in the Grinsted et al. [1] analysis is an accurate characterization of an individual spider's stable behavioural phenotype. If we have no confidence that ‘boldness’ is repeatable in this population of spiders then two out of the three a priori hypotheses in Grinsted et al. [1] can no longer be definitively tested. The first (‘variation in individual personality trait values can predict task differentiation…’) and third (‘standardized personality assays, devoid of social context, can be used as predictors of individual behaviours in natural settings…’) hypotheses, as stated in the final paragraph of the introduction, both rely on the existence of ecologically-relevant levels of personality variation in these spiders. Without evidence of reliable repeatabilities in boldness, the findings of this study are reduced to a correlation of a single behavioural measure in captivity (that could reflect some unmeasured, ecologically-arbitrary influence) with behavioural outcomes in the field.On this basis, we are retracting the paper.Footnotes© 2021 The Author(s)Published by the Royal Society. All rights reserved.

Safety and Immunogenicity of a Stable, Cold-Adapted, Temperature-Sensitive/Conditional Lethal Enterovirus A71 in Monkey Study.

Enterovirus A71 (EV-A71) and coxsackievirus A16 (CA16) are major etiological agents of hand foot and mouth disease (HFMD) in children, which may result in fatal neurological complications. The development of safe, cost effective vaccines against HFMD, especially for use in developing countries, is still a top public health priority. We have successfully generated a stable, cold-adapted, temperature sensitive/conditional lethal EV-A71 through adaptive culturing in Vero cells at incrementally lower cultivation temperatures. An additional 40 passages at an incubation temperature of 28 °C, and a temperature reversion study at an incubation temperature of 37 °C and 39.5 °C, reveals the virus’s phenotypic and genetic stability at the predefined culture conditions. Six unique mutations (two in noncoding regions and four in nonstructural protein-coding genes) in combination may have contributed to its stable phenotype and inability to fully revert to its original wild phenotype. The safety and immunogenicity of this stable, cold-adapted, temperature sensitive/conditional lethal EV-A71 was performed in six monkeys. None of the inoculated monkeys developed any obvious clinical illness except one which developed a transient spike of fever. No gross postmortem lesion or abnormal histological finding was noted for all monkeys at autopsy. No virus was reisolated although EV-A71 specific RNA was detected in serum samples collected on both day 4 and day 8 postinoculation. Only EV-A71 RNA and viral antigen were detected in the spleen homogenate and peripheral blood mononuclear cells, respectively, collected on day 4. The two remaining monkeys developed good humoral immune response on day 14 and day 30 post-inoculation.

Nonconservative integration and diversity of a new family of integrative and conjugative elements associated with antibiotic resistance in zoonotic pathogen Streptococcus suis

Macrolide and tetracycline resistance in streptococci is mainly caused by acquisition of integrative and conjugative elements (ICEs) of the ICESa2603 family carrying erm(B) and tet(O). But the characteristics about the transferability and physiological consequences of ICEs with triplet serine integrases are still rare. This study tested the transferability of ICESsuYZDH1_SSU0877, a novel erm(B)- and tet(O)-carrying ICESa2603 family-like ICE with triplet serine integrases, and evaluated the physiological consequences after ICE transferred and integrated into recipient. The prevalence of ICESsuYZDH1-like ICEs in S. suis was analyzed based on 1334 genomic sequences available in GenBank and examined in 330 clinical isolates in China. Nonconservative transfer was observed by integrating of ICESsuYZDH1 into SSU1797 gene besides the primary SSU0877 site. Imperfect direct repeats of 2-/4-nt (5’-TC-3’/5’-TCCC-3’) and (5’-GC-3’/5’-TCCC-3’) were observed at SSU0877 and SSU1797 sites, respectively. The transconjugant suffered a weak fitness cost with stunted growth and less competition with recipient strain. Successive passages indicate the ICESsuYZDH1 could be persist and endued stable resistant phenotype. Comprehensive analysis of the ICESsuYZDH1-like ICEs from both public genome database and our clinical isolates revealed the widespread and diversity of the ICEs by integration at the sites of SSU0877, SSU0468, SSU1262, and SSU1797. The ICESsuYZDH1-like ICEs could stably co-exist within the host chromosome at more than one attachment sites, which is probably mediated by the triplet serine integrases. Nonconservative integration and diversity of the ICESsuYZDH1 family of ICEs might have contributed to the evolution of ICEs and the dissemination of macrolide and tetracycline resistance in S. suis.

Mining favorable alleles for rice sheath blight resistance by association mapping

Sheath blight is a serious rice disease that causes great yield losses worldwide. Sheath blight resistance is controlled by genes with minor effect. It is an urgent need to identify the resistance genes in rice germplasm and utilize them in breeding. The purpose of this study is to identify the sheath blight-resistant alleles in different rice germplasm by association mapping and explore the potential of the resistance alleles in utilization. A total of 273 rice genotypes were inoculated and evaluated for sheath blight resistance in the field for 5 years, and 158 genotypes with relatively stable disease resistance phenotype were chosen for association mapping. A wide range of phenotypic variation for sheath blight resistance was observed in the 158-genotype population. Structure analyses divided the 158 genotypes into 4 subgroups, which was consistent with principal component analysis and cluster analysis. Association mapping performed using mixed linear model with 213 markers identified 14 resistance loci, locating on all chromosomes except chromosome 6, 10 and 12. Three novel resistance loci (D704, D855, and D905) which were different from the previously reported were detected. Thirty-eight alleles were detected at the 14 loci, among which favorable alleles at 11 loci showed consistent resistance across 5 years. It was found that the favorable resistance alleles can be pyramided to improve sheath blight resistance. These results enhance the understanding of how different resistance alleles at multiple loci in regulating sheath blight resistance, and provide markers for sheath blight resistance breeding in rice.

Translational Control during Cellular Senescence.

Senescence is a state of long-term cell cycle arrest that arises in cells that have incurred sublethal damage. While senescent cells no longer replicate, they remain metabolically active and further develop unique and stable phenotypes that are not present in proliferating cells. On one hand, senescent cells increase in size, maintain an active mTORC1 complex, and produce and secrete a substantial amount of inflammatory proteins as part of the senescence-associated secretory phenotype (SASP). On the other hand, these progrowth phenotypes contrast with the p53-mediated growth arrest typical of senescent cells that is associated with nucleolar stress and an inhibition of rRNA processing and ribosome biogenesis. In sum, translation in senescent cells paradoxically comprises both a global repression of translation triggered by DNA damage and a select increase in the translation of specific proteins, including SASP factors.

Protective effect of estrogen receptors (ERα/β) against the intervertebral disc degeneration involves activating CCN5 via the promoter.

Due to the decrease of estrogen and estrogen receptor (ER) in postmenopausal women, they have a higher risk of intervertebral disc degeneration (IDD) than men. This study aims to explore how ERα and ERb interact with CCN5 and protect IDD. We used Chromatin immunoprecipitation (ChIP) and Luciferase reporter assay to determine whether the ERα/b protein binds to CCN5 promoter and activates its expression. We used TNF-α to induce nucleus pulposus (NP) cell degeneration to simulate the IDD process. The change of the expression of ERα/β and CCN5 was measured in the degenerated NP cells. To understand the function of ERα/β in the NP cells degeneration, we upregulated the ERα/b gene expression by vector transfection or 17b-estradiol (E2) stimulation. Besides, we also used the CCN5 gene-silenced NP cells by siRNA transfection as a comparison to determine the role of CCN5. We tested the cell proliferation and principal components of the extracellular matrix (ECM) to value the degree of NP cell degeneration. ERα and ERβ protein can bind to the same promoter regions of CCN5 and activate its expression, respectively. TNF-α degraded NP cells with a reduction of cell proliferation, collagen II, ACAN, ERα, ERβ, and CCN5 expression, and increased collagen I/III, and MMP-13 expression. Upregulated ERα or ERβ resulted in the maintains of CCN5 and alleviated the NP cell degeneration. Besides, 17β-E2 supplement increased the ERα, ERβ, and CCN5 expression, as well as stable NP cells phenotype. However, it was partly abolished by the silencing of CCN5. Upregulation of ERα and ERβ protects the NP cell degeneration during IDD through the activation of CCN5 by binding to its promoter.

Effects of Nicorandil on Inflammation, Apoptosis and Atherosclerotic Plaque Progression.

Nicorandil, a balanced vasodilator, is used in the second-line therapy of angina pectoris. In this study, we aimed to illuminate the effects of nicorandil on inflammation, apoptosis, and atherosclerotic plaque progression. Twenty-five LDL-R -/- mice were fed a high-fat diet for 14 weeks. After 6 weeks mice were randomly allocated to treatment with nicorandil (10 mg/kg/day) or tap water. Nicorandil treatment led to a more stable plaque phenotype, displaying an increased thickness of the fibrous cap (p = 0.014), a significant reduction in cholesterol clefts (p = 0.045), and enhanced smooth muscle cell content (p = 0.009). In endothelial cells nicorandil did not reduce the induction of adhesion molecules or proinflammatory cytokines. In H2O2 challenged endothelial cells, pretreatment with nicorandil significantly reduced the percentage of late apoptotic/necrotic cells (p = 0.016) and the ratio of apoptotic to living cells (p = 0.036). Atherosclerotic lesions of animals treated with nicorandil exhibited a significantly decreased content of cleaved caspase-3 (p = 0.034), lower numbers of apoptotic nuclei (p = 0.040), and reduced 8-oxogunanine staining (p = 0.039), demonstrating a stabilizing effect of nicorandil in established atherosclerotic lesions. We suggest that nicorandil has a positive effect on atherosclerotic plaque stabilization by reducing apoptosis.

Chronic Hyperglycemia Drives Functional Impairment of Lymphocytes in Diabetic INSC94Y Transgenic Pigs.

People with diabetes mellitus have an increased risk for infections, however, there is still a critical gap in precise knowledge about altered immune mechanisms in this disease. Since diabetic INSC94Y transgenic pigs exhibit elevated blood glucose and a stable diabetic phenotype soon after birth, they provide a favorable model to explore functional alterations of immune cells in an early stage of diabetes mellitus in vivo. Hence, we investigated peripheral blood mononuclear cells (PBMC) of these diabetic pigs compared to non-diabetic wild-type littermates. We found a 5-fold decreased proliferative response of T cells in INSC94Y tg pigs to polyclonal T cell mitogen phytohemagglutinin (PHA). Using label-free LC-MS/MS, a total of 3,487 proteins were quantified, and distinct changes in protein abundances in CD4+ T cells of early-stage diabetic pigs were detectable. Additionally, we found significant increases in mitochondrial oxygen consumption rate (OCR) and higher basal glycolytic activity in PBMC of diabetic INSC94Y tg pigs, indicating an altered metabolic immune cell phenotype. Thus, our study provides new insights into molecular mechanisms of dysregulated immune cells triggered by permanent hyperglycemia.

Early stability and late random tumor progression of a HER2-positive primary breast cancer patient-derived xenograft

We established patient-derived xenografts (PDX) from human primary breast cancers and studied whether stability or progressive events occurred during long-term in vivo passages (up to 4 years) in severely immunodeficient mice. While most PDX showed stable biomarker expression and growth phenotype, a HER2-positive PDX (PDX-BRB4) originated a subline (out of 6 studied in parallel) that progressively acquired a significantly increased tumor growth rate, resistance to cell senescence of in vitro cultures, increased stem cell marker expression and high lung metastatic ability, along with a strong decrease of BCL2 expression. RNAseq analysis of the progressed subline showed that BCL2 was connected to three main hub genes also down-regulated (CDKN2A, STAT5A and WT1). Gene expression of progressed subline suggested a partial epithelial-to-mesenchymal transition. PDX-BRB4 with its progressed subline is a preclinical model mirroring the clinical paradox of high level-BCL2 as a good prognostic factor in breast cancer. Sequential in vivo passages of PDX-BRB4 chronically treated with trastuzumab developed progressive loss of sensitivity to trastuzumab while HER2 expression and sensitivity to the pan-HER tyrosine kinase inhibitor neratinib were maintained. Long-term PDX studies, even though demanding, can originate new preclinical models, suitable to investigate the mechanisms of breast cancer progression and new therapeutic approaches.

Apoptotic Ablation of Platelets Reduces Atherosclerosis in Mice With Diabetes.

People with diabetes are at a significantly higher risk of cardiovascular disease, in part, due to accelerated atherosclerosis. Diabetic subjects have increased number of platelets that are activated, more reactive, and respond suboptimally to antiplatelet therapies. We hypothesized that reducing platelet numbers by inducing their premature apoptotic death would decrease atherosclerosis. Approach and Results: This was achieved by targeting the antiapoptotic protein Bcl-xL (B-cell lymphoma-extra large; which is essential for platelet viability) via distinct genetic and pharmacological approaches. In the former, we transplanted bone marrow from mice carrying the Tyr15 to Cys loss of function allele of Bcl-x (known as Bcl-xPlt20) or wild-type littermate controls into atherosclerotic-prone Ldlr+/- mice made diabetic with streptozotocin and fed a Western diet. Reduced Bcl-xL function in hematopoietic cells significantly decreased platelet numbers, exclusive of other hematologic changes. This led to a significant reduction in atherosclerotic lesion formation in Bcl-xPlt20 bone marrow transplanted Ldlr+/- mice. To assess the potential therapeutic relevance of reducing platelets in atherosclerosis, we next targeted Bcl-xL with a pharmacological strategy. This was achieved by low-dose administration of the BH3 (B-cell lymphoma-2 homology domain 3) mimetic, ABT-737 triweekly, in diabetic Apoe-/- mice for the final 6 weeks of a 12-week study. ABT-737 normalized platelet numbers along with platelet and leukocyte activation to that of nondiabetic controls, significantly reducing atherosclerosis while promoting a more stable plaque phenotype. These studies suggest that selectively reducing circulating platelets, by targeting Bcl-xL to promote platelet apoptosis, can reduce atherosclerosis and lower cardiovascular disease risk in diabetes. Graphic Abstract: A graphic abstract is available for this article.

Targeted De-Methylation of the FOXP3-TSDR Is Sufficient to Induce Physiological FOXP3 Expression but Not a Functional Treg Phenotype.

CD4+ regulatory T cells (Tregs) are key mediators of immunological tolerance and promising effector cells for immuno-suppressive adoptive cellular therapy to fight autoimmunity and chronic inflammation. Their functional stability is critical for their clinical utility and has been correlated to the demethylated state of the TSDR/CNS2 enhancer element in the Treg lineage transcription factor FOXP3. However, proof for a causal contribution of the TSDR de-methylation to FOXP3 stability and Treg induction is so far lacking. We here established a powerful transient-transfection CRISPR-Cas9-based epigenetic editing method for the selective de-methylation of the TSDR within the endogenous chromatin environment of a living cell. The induced de-methylated state was stable over weeks in clonal T cell proliferation cultures even after expression of the editing complex had ceased. Epigenetic editing of the TSDR resulted in FOXP3 expression, even in its physiological isoform distribution, proving a causal role for the de-methylated TSDR in FOXP3 regulation. However, successful FOXP3 induction was not associated with a switch towards a functional Treg phenotype, in contrast to what has been reported from FOXP3 overexpression approaches. Thus, TSDR de-methylation is required, but not sufficient for a stable Treg phenotype induction. Therefore, targeted demethylation of the TSDR may be a critical addition to published in vitro Treg induction protocols which so far lack FOXP3 stability.

Macrophage targeted theranostic strategy for accurate detection and rapid stabilization of the inflamed high-risk plaque.

Rationale: Inflammation plays a pivotal role in the pathogenesis of the acute coronary syndrome. Detecting plaques with high inflammatory activity and specifically treating those lesions can be crucial to prevent life-threatening cardiovascular events.Methods: Here, we developed a macrophage mannose receptor (MMR)-targeted theranostic nanodrug (mannose-polyethylene glycol-glycol chitosan-deoxycholic acid-cyanine 7-lobeglitazone; MMR-Lobe-Cy) designed to identify inflammatory activity as well as to deliver peroxisome proliferator-activated gamma (PPARγ) agonist, lobeglitazone, specifically to high-risk plaques based on the high mannose receptor specificity. The MMR-Lobe-Cy was intravenously injected into balloon-injured atheromatous rabbits and serial in vivo optical coherence tomography (OCT)-near-infrared fluorescence (NIRF) structural-molecular imaging was performed.Results: One week after MMR-Lobe-Cy administration, the inflammatory NIRF signals in the plaques notably decreased compared to the baseline whereas the signals in saline controls even increased over time. In accordance with in vivo imaging findings, ex vivo NIRF signals on fluorescence reflectance imaging (FRI) and plaque inflammation by immunostainings significantly decreased compared to oral lobeglitazone group or saline controls. The anti-inflammatory effect of MMR-Lobe-Cy was mediated by inhibition of TLR4/NF-κB pathway. Furthermore, acute resolution of inflammation altered the inflamed plaque into a stable phenotype with less macrophages and collagen-rich matrix.Conclusion: Macrophage targeted PPARγ activator labeled with NIRF rapidly stabilized the inflamed plaques in coronary sized artery, which could be quantitatively assessed using intravascular OCT-NIRF imaging. This novel theranostic approach provides a promising theranostic strategy for high-risk coronary plaques.

Widely targeted metabolomic analyses of albino tea germplasm ‘Huabai 1’ and ‘Baiye 1’

‘Huabai 1’ is a novel albino tea germplasm that has a stable albino phenotype and prolonged albino period. In this study, widely targeted metabolomics was employed to analyze the metabolic profile of ‘Huabai 1’, and compare metabolites between albino shoots of ‘Huabai 1’ (HBW) and albino shoots of the broadest cultivated cultivar ‘Baiye 1’ (BYW). A total of 359 differential metabolites were identified in ‘Huabai 1’. Comparative analysis of differential metabolites revealed that 83 and 161 differential metabolites were specific for HBW and BYW, respectively. The phenylpropanoid and flavonoids biosynthesis are the key metabolic pathways, contributing to the color change in ‘Huabai 1’, while the purine metabolism and lipid-related metabolites may play important roles in regulating albinism in ‘Baiye 1’. Comparative analysis of amino acids and catechins revealed that the umami taste of albino tea may be due to the internal balance of amino acid components and the decrease in phenol-ammonia ratio. In addition, lipid-related metabolites, the main contributors to the aroma of tea, were significantly enriched in BYW, while, HBW possess higher antioxidative components than that of BYW, such as apigenin and genistein etc., suggesting that the ‘Huabai 1’ may have high prospects in commercialization and future breeding programs.