Biomimetic Synthesis of Moschamine-Related Indole Alkaloids via Iron-Catalyzed Selectively Oxidative Radical Coupling

An iron-catalyzed oxidative radical coupling reaction was developed to selectively construct indolofuran or bisphenolic indole cores, which exist in two types of moschamine-related indole alkaloids. Both (+)-decursivine and 4,4″-bis­(<i>N</i>-feruloyl)­serotonin were biomimetically synthesized by using coupling reactions. The proposed reassignment of the structure of montamine as 4,4″-bis­(<i>N</i>-feruloyl)­serotonin was excluded

MAb 8E6G4 competes with b96.11 for binding to GAD65.

<p><b>A:</b> GAD65 was translated <i>in vitro</i> using rabbit reticulocyte lysate. Different amounts (0–40 µl, corresponding to 0–160 ng) of recombinant GAD65 were incubated with 1.4 µg/ml MAb 8E6G4 and subsequently analyzed for binding to b96.11 by ELISA. Rabbit reticulocyte lysate alone was used as a negative control (open squares). Binding in the absence of competitor was set as 100%. <b>B:</b> Binding of monoclonal GAD65Ab b96.11 (black squares), b78 (open squares), and N-GAD65mAb (open circles) to radiolabeled GAD65 in the presence of the indicated concentrations of MAb 8E6G4 was determined. Binding is reported at percent binding, binding of the respective antibody to radiolabeled GAD65 in the absence of MAb 8E6G4 is set as 100%.</p

Table_1_Association between blood chromium and hepatic steatosis assessed by liver ultrasound transient elastography: National Health and Nutrition Examination Survey 2017–2020.docx

BackgroundHepatic steatosis is a significant pathological feature of fatty liver disease (FLD) which is widely spread with no effective treatment available. Previous studies suggest that chromium (Cr) intake reduces lipid deposition in the liver in animals. However, the connection between blood Cr and hepatic steatosis among humans remains inconclusive.MethodsUsing the data from the National Health and Nutrition Examination Survey (NHANES) 2017–2020, we performed a cross-sectional analysis, including 4,926 participants. The controlled attenuation parameter (CAP) measured by the vibration controlled transient elastography (VCTE) was used to evaluate the degree of liver steatosis. Weighted univariate regression, multivariate linear regression, smooth fitting curves and subgroup analysis were used. In addition, we carried out trend tests, multiple interpolations, and interaction analyses to conduct sensitivity analyses.ResultsAfter adjusting with various covariables, multivariate linear regression analysis demonstrated a significant negative correlation between blood Cr and CAP [β (95% CI) = −5.62 (−11.02, −0.21)]. The negative correlation between blood Cr and CAP was more significant in the males, 50–59 years, overweight, hypercholesterolemia, HDL-C ≥ 65 mg/dL, HbA1c (5.70–6.10 %), HOMA-IR (0.12–2.76), total bilirubin (0.30–0.40 mg/dL), ever alcohol consumption subjects. Of note, the relationships between blood Cr and CAP followed a U-shaped curve in the smokers and non-smokers, with blood Cr thresholds of 0.48, 0.69 μg/L, respectively.ConclusionsThere is an independently negative correlation between blood Cr and hepatic steatosis in American. Our study provides clinical researchers with a new insight into the prospective prevention of hepatic steatosis.</p

MAb 8E6G4 binds specifically to b96.11.

<p><b>A:</b> Purified MAb 8E6G4, supernatant of hybridoma 8E6G4, or BSA were analyzed for binding by b96.11 by Dot blot. From left: supernatant of hybridoma 8E6G4, purified MAb 8E6G4, and BSA. <b>B:</b> Binding of MAb 8E6G4 and supernatant of hybridoma 8E6G4 to b96.11 was analyzed by immunoprecipitation followed by Western blot analysis. Binding of MAb 8E6G4 to control antibody hLF served as negative control. From left: supernatant of hybridoma 8E6G4 with b96.11, MAb 8E6G4 with b96.11, and MAb 8E6G4 with hLF. <b>C:</b> MAb 8E6G4 binds to b96.11 with high affinity. Dissociation constant was calculated by non-competing ELISA. Different concentrations of b96.11 were tested for binding by MAb 8E6G4. Kd of each concentration was calculated. The Kd was calculated as follows: Kd = 2(nKd′-Kd)/(n-1), where n = [b96.11]′/[b96.11]. The average Kd calculated was 480.4 pM.</p

MAb 8E6G4 binds GAD65Ab present in human sera.

<p><b>A:</b> Human sera obtained from GAD65Ab-negative healthy individuals (circles) (n = 40), GAD65Ab-negative T2D patients (squares) (n = 22), and T1D patients (diamonds) (n = 10), were analyzed for binding to MAb 8E6G4 by ELISA. Binding of human sera to three mouse monoclonal antibodies specific to human HGF is shown as a negative control (open symbols). Median binding is indicated. <b>B:</b> Human sera obtained from healthy individuals (n = 40) (GAD65Ab-negative in RBA) were tested for presence of masked GAD65Ab. Sera were precipitated on immobilized MAb 8E6G4, and subsequently eluted. Elutions were tested for presence of GAD65Ab by RBA. GAD65Ab titers of sera prior to precipitation (black circles) and elutions (open circles) are presented as GAD65Ab index. Median binding is indicated.</p

The histopathology of pancreatic islets at onset of diabetes or at 40 weeks of age.

<p>Representative images of pancreatic islets from animals injected with 50 µg (A) and 100 µg (B) MAb 8E6G4 IgG, or control animals (C). Pancreatic tissues were sectioned and stained with hematoxylin and eosin.</p

Degree of insulitis in NOD mice.

<p>Islets obtained from animals injected with MAb 8E6G4 IgG (50 and 100 µg), control IgG, and PBS were scored as normal islets (score 1), perivascular/periductal infiltration (score 2), peri-insulitis (score 3), mild insulitis (<25% of the islet infiltrated; score 4), and severe insulitis (more than 25% of the islet infiltrated, score 5). The mean score for each group was calculated by dividing the total score by the number of islets scored.</p

Cumulative incidence rate of diabetes development.

<p>NOD mice were injected with 50 and 100 µg MAb 8E6G4, presented as black circles and squares, respectively. Control animals were injected with PBS (white circles) or control mouse monoclonal antibody (white squares). Animals were weekly monitored for diabetes development. Cumulative incidence rate of diabetes development is presented as percentage.</p