One-Pot Synthesis and Structure–Property Relationship of Aminomaleimides: Fluorescence Efficiencies in Monomers and Aggregates Easily Tuned by Switch of Aryl and Alkyl

Organic fluorophores have attracted great interest owing to their wide applications. They usually contain an electron-conjugated system with an aromatic moiety and show high emission in dilute solutions but weaker or even no emission upon aggregation. Here, a simple one-pot, three-component reaction (3CR) (method I) for the synthesis of various di- and monosubstituted aminomaleimides (DAMIs and MAMIs) has been developed, and the reported 3CR (method II) has been found to be efficient only for the synthesis of MAMIs with R² = alkyl. Twelve AMIs were designed and synthesized for investigation of the influence of structures on their optical properties in monomers and aggregates. It was found that alkyl MAMIs, alkyl DAMIs, and aryl AMIs/DAMIs show very different fluorescence efficiencies in different solvents, and only MAMIs with butyl and oleyl show high emissions in powders similar to those in nonpolar solutions. Single-crystal structures indicate that their fluorescence efficiencies in aggregates mainly correlate with molecular packing modes. The efficient synthesis method, the sensitive fluorescence on–off response to protic solvents or polar solvents, and the unusual high emissions of AMI without any aromatic moiety in both monomer and aggregates are expected to attract great interest in the fields of application and theory

Table_1_Assessment of biochemical outcomes in patients with primary aldosteronism after adrenalectomy based on CT scan diagnosis of unilateral adenoma without adrenal vein sampling.doc

PurposeThe purpose of this study was to assess the surgical outcomes of patients with primary aldosteronism when surgery was based only on CT finding of unilateral adenoma without adrenal vein sampling (AVS).MethodsThis is a retrospective review of the records of patients who had undergone retroperitoneal laparoscopic adrenalectomy for primary aldosteronism based on CT scan finding of unilateral adenoma and had a follow-up of at least 6–12 months from January 2012 to December 2020 in a single center; decision for adrenalectomy was based on CT scan, and AVS was not used. The clinical and biochemical outcomes were accessed using the standardized primary aldosteronism surgical outcome (PASO) criteria. Patient’s demographics and preoperative factors were analyzed to assess for independent predictor of surgical success.ResultsAccording to the PASO criteria, 172 patients finally enrolled in the training dataset, and 20 patients enrolled in the validation dataset. In the training dataset, complete clinical success was achieved in 71 patients (41.3%), partial success in 87 (50.6%), and absent success in 14 (8.1%). Biochemical outcomes showed that 151 patients (87.8%) were completely cured, 14 patients (8.1%) got a partial biochemical success, and an absent biochemical success was found in seven patients (4.1%). Multivariate logistic regression analysis showed that age, body mass index (BMI), tumor size, mean arterial pressure (MAP), and serum potassium were the most independent factors for incomplete biochemical success. Based on the results of statistical analysis, our study constructed a nomogram prognostic evaluation model for patients after unilateral primary aldosterone surgery.ConclusionsLaparoscopic adrenalectomy for patients with primary aldosteronism base on CT scan finding of a unilateral adenoma without AVS had a high rate of complete biochemical cure at 12 months. Risk factors for incomplete biochemical success include age, BMI, tumor size, MAP, and serum potassium. Our study constructed a nomogram prognostic evaluation model for patients after unilateral primary aldosterone surgery. The nomogram accurately and reliably predicted the incomplete biochemical success.</p

Additional file 2 of Ovulatory signal-triggered chromatin remodeling in ovarian granulosa cells by HDAC2 phosphorylation activation-mediated histone deacetylation

Additional file 2: Table S1. Oligonucleotides used for qPCR or siRNA. Table S2. Reagents or commercial kits used in this study. Table S3. The DEG list of RNA-seq data (H4 vs H0)

Additional file 1 of Ovulatory signal-triggered chromatin remodeling in ovarian granulosa cells by HDAC2 phosphorylation activation-mediated histone deacetylation

Additional file 1. Figure S1. The Dynamics of H3K27Ac Levels during Follicular Growth and Ovulation. Mice were treated with pregnant mare serum gonadotrophin (PMSG) to stimulate follicle growth or human chorionic gonadotropin (hCG) to trigger ovulation. Ovaries were collected at the indicated timepoints for immunofluorescence. The representative images showing the H3K27Ac (red) levels with DAPI (blue) co-stained for visualization of nucleus (n=3). Scale bar=100um. Figure S2. The H3K27Ac ChIP-seq Analysis for H3K27Ac-gain Genes after Ovulation Signal Induction. (A) Pie charts represent the ratio of stable H3K27Ac-enriched peaks, H3K27-loss peaks and H3K27-gain peakss (de novo H3K27Ac-deposited peaks and H3K27Ac-increased peaks). (B) Genome browser snapshot shows Prss56 is one of de novo H3K27Ac-gain genes after hCG induction. (C) Gene Ontology (GO) analysis shows the biological process (BP) of de novo H3K27Ac-gain genes after hCG induction. (D) Bar charts representing the top enriched KEGG pathway of the ovulatory specific genes 4 h post hCG. (E) The HOMER known and de novo motif analysis for the H3K27Ac-gain peaks to predict transcriptional factors. Figure S3. Semiquantitative analysis of protein levels in Fig. 3. The western blot band intensities of Fig. 3B were measured with ImageJ software. HDAC1 and p-HDAC2 protein levels were normalized to Histone H3 levels. Data were expressed as mean±SD. P value was determined by two-way ANOVA followed by Tukey’s post-test. ** P<0.01.Figure S4. Depletion or inhibition of HDAC2 Causes Increased H3K27Ac Levels and Affects COC expansion. (A) Western blot analysis and quantification of HDAC2 and H3K27Ac levels in siNC and siHdac2 group. Primary granulosa cells were transfected of negative control (siNC) or Hdac2 siRNAs (siHdac2) for 36 h followed by treatment of forskolin (FSK, 10 μM) and phorbol 12-myristate 13-acetate (PMA, 20 nM) for indicated time length. All the protein levels were normalized to Histone H3 levels. The HDAC2 bands’ intensities were compared with siNC and siHdac2 groups in C. The bands’ intensities of H3K27Ac were compared among FSK/PMA non-treated group (0h) and FSK/PMA treated groups (0.5h, 1h, 2h and 4h). (B) Semiquantitative analysis of p-HDAC2 and H3K27Ac levels in Fig. 4A. The western blot band intensities of Fig. 4A were measured with ImageJ software. All protein levels were normalized to Histone H3 levels. The siHDAC2 bands’ intensities were compared with siNC bands’ intensities. (C) Comparation of COCs expansion areas in Fig. 4B. The areas of COCs expansion in siNC and siHDAC2 groups were measured with ImageJ software. (D) Quantitative analysis of H3K27Ac, H3K9Ac and H4K16Ac levels in Fig. 5B. All the protein levels were normalized to histone H3 levels. The FK228 bands’ intensities were compared with control bands’ intensities. All data in A-D were expressed as mean±SD. P value was determined by two-way ANOVA followed by Tukey’s post-test. ns. means no significance. * P<0.05, ** P<0.01. Figure S5. Inhibition of HDAC1/2 Hinders Cumulus Expansion and Ovulation. Hematoxylin and eosin (HE) staining results showing FK228, a HDAC1/2 inhibitor, blocks cumulus expansion and cause follicle atresia. Mice pretreated PMSG for 44 h were injected with DMSO or FK228, and 4 h later followed by hCG injection to induce ovulation (n=12). Ovaries were collected at 0 h, 4 h and 8 h post-hCG in mice of control and FK228 group. Scale bar, 50 um. Figure S6. Semiquantitative analysis of CK2α, p-HDAC2, H3K27Ac and p-ERK1/2 levels in Fig. 6A and C. The western blot band intensities were measured with ImageJ software. CK2α, p-HDAC2 and H3K27Ac protein levels were normalized to Histone H3 levels and p-ERK1/2 levels were normalized to ERK1/2. The TBB bands’ intensities were compared with control bands’ intensities. Data were expressed as mean±SD. P value was determined by two-way ANOVA followed by Tukey’s post-test. ** P<0.0