Journal: EMBO Reports

Article Title: miRNA-target complementarity in cnidarians resembles its counterpart in plants

doi: 10.1038/s44319-024-00350-z

Figure Lengend Snippet: Reagents and tools table

Article Snippet: In vitro transcription was conducted with HighYield T7 Cap 1 AG (3‘-OMe) mRNA Synthesis Kit (m5CTP) (Jena Bioscience, Germany) using 800 ng of amplified template followed by Turbo DNase treatment (Thermo Fisher Scientific) by incubation with 1 µl of DNase for 30 min at 37 °C twice sequentially.

Techniques: Sequencing, shRNA, SYBR Green Assay, Protease Inhibitor, Bicinchoninic Acid Protein Assay, Enzyme-linked Immunosorbent Assay, Plasmid Preparation, Magnetic Beads, Multiplex Assay, Sensitive Assay, Software, Imaging, Spectrophotometry, Real-time Polymerase Chain Reaction

Journal: International Journal of Molecular Sciences

Article Title: Liver-Specific Overexpression of Prostasin Attenuates High-Fat Diet-Induced Metabolic Dysregulation in Mice

doi: 10.3390/ijms22158314

Figure Lengend Snippet: Serum PRSS8 levels was negatively associated with the presence of type 2 diabetes and increased intima media thickness. ( a ) The ELISA of human PRSS8 levels in the serum of the nondiabetes mellitus (non-DM) and type 2 diabetes mellitus (T2DM) participants (n = 39–40 per group). Values are shown as the mean ± SEM; * p -value < 0.05 ( t test). ( b ) The maximum intima media thickness (max IMT) measured by carotid ultrasonography in T2DM. Values are shown as the mean ± SEM; ** p -value < 0.01 ( t test). ( c ) Homeostatic model assessment beta cell function (HOMA-β) in non-DM was shown (Pearson product-moment correlation coefficient).

Article Snippet: Measurement of serum fasting PRSS8 levels in WT/LTg mice was used by Human Prostasin ELISA Kit (BOSTER Biological Technology Co., Ltd. Pleasanton, CA, USA).

Techniques: Enzyme-linked Immunosorbent Assay, Cell Function Assay

Journal: Medicine

Article Title: Role of serum CAP1 protein in the diagnosis of patients with first-time acute myocardial infarction

doi: 10.1097/MD.0000000000034700

Figure Lengend Snippet: The expression of CAP1 is upregulated in serum of patients with first-time AMI. (A) ELISA analysis of the concentration of CAP1 protein in serum samples from 70 patients with first-time AMI and 62 healthy controls. (B) Analysis of the CAP1 protein expression at 0, 6, 12, 24, 48 hours and 7 d of the onset of chest pain in patients with first-time AMI. (C) Pearson correlation analysis showed CAP1 protein expression was positively correlated with cTnI level in patients with first-time AMI. (D) A positive correlation was found between CAP1 protein expression and ox-LDL level in patients with first-time AMI. The results were shown as the means ± standard deviations. * P < .05, ** P < .01, *** P < .001. AMI = acute myocardial infarction, CAP1 = cyclase associated actin cytoskeleton regulatory protein 1, cTnI = cardiac troponin I, ELISA = enzyme-linked immunosorbent assay, ox-LDL = oxidized low-density lipoprotein.

Article Snippet: The concentration of CAP1 protein in serum was detected with human CAP1 ELISA assay kit from CUSABIO (Wuhan, China).

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Concentration Assay

Journal: Medicine

Article Title: Role of serum CAP1 protein in the diagnosis of patients with first-time acute myocardial infarction

doi: 10.1097/MD.0000000000034700

Figure Lengend Snippet: Comparisons of the AUC by serum CAP1 protein and cTnT level for the diagnosis of patients with first-time AMI. The ROC curve analysis was carried out to detect CAP1 as biomarker for differentiating patients with first-time AMI from healthy control. (A) The AUC was 0.794 (95% CI: 0.718–0.869) for CAP1. (B) The AUC was 0.720 (95% CI: 0.633–0.807) for cTnT. (C) The AUC was 0.864 (95% CI: 0.802–0.925) for CAP1 and cTnT combination. AMI = acute myocardial infarction. AUC = area under curve, CAP1 = cyclase associated actin cytoskeleton regulatory protein 1, ROC = receiver operating characteristic.

Article Snippet: The concentration of CAP1 protein in serum was detected with human CAP1 ELISA assay kit from CUSABIO (Wuhan, China).

Techniques: Biomarker Discovery, Control

Journal: Medicine

Article Title: Role of serum CAP1 protein in the diagnosis of patients with first-time acute myocardial infarction

doi: 10.1097/MD.0000000000034700

Figure Lengend Snippet: The effect of CAP1 on cell proliferation, inflammation, and NF-κB pathway activity in ox-LDL induced HUVEC. (A) 5-BrdU assay analysis of the proliferative ability of ox-LDL induced HUVEC by CAP1 knockdown and overexpression. (B) ELISA assay analysis of IL-1β, TNF-α, MCP-1, and IL-6 expression in ox-LDL induced HUVEC by overexpression of CAP1. (C) CAP1 knockdown by CAP1-siRNA inhibited the expression of IL-1β, TNF-α, MCP-1, and IL-6. (D) Luciferase reporter gene assay analysis of NF-κB luciferase activity in ox-LDL induced HUVEC by CAP1 knockdown and overexpression. The results were shown as the means ± standard deviations. * P < .05, ** P < .01, *** P < .001, ns P > 0.05. BrdU = bromodeoxyuridine, CAP1 = cyclase associated actin cytoskeleton regulatory protein 1, ELISA = enzyme-linked immunosorbent assay, HUVEC = human umbilical vein endothelial cells, ox-LDL = oxidized low density lipoprotein.

Article Snippet: The concentration of CAP1 protein in serum was detected with human CAP1 ELISA assay kit from CUSABIO (Wuhan, China).

Techniques: Activity Assay, BrdU Staining, Knockdown, Over Expression, Enzyme-linked Immunosorbent Assay, Expressing, Luciferase, Reporter Gene Assay

Journal: Nature Communications

Article Title: Amplified fluorogenic immunoassay for early diagnosis and monitoring of Alzheimer’s disease from tear fluid

doi: 10.1038/s41467-023-43995-5

Figure Lengend Snippet: Sixty human tear fluid samples were utilized in two cohorts: a discovery cohort and a verification cohort. The discovery cohort was comprised of tear samples from 7 healthy controls (HC), 7 patients with mild cognitive impairment (MCI), and 7 patients with Alzheimer’s disease (AD) for a total of 21 tear samples. Proteomic analysis using mass spectrometry identified adenylyl cyclase-associated protein 1 (CAP1) as a promising potential biomarker. The verification cohort consisted of 14 HC, 15 MCI patients, and 10 AD patients, with a total of 39 tear samples. A self-assembled nanoparticle-mediated fluorescence immunoassay (SNAFIA) was applied to detect the target protein, CAP1, in human tear fluid samples. The presence of the CAP1 protein generated a sandwich immunocomplex with an antibody-conjugated magnetic nanoparticle (Ab-MNP) and an antibody-conjugated polymeric nanoprobe (Ab-PNP), enabling the discrimination of the three groups of participants based on the analysis of the fluorescence signals from the SNAFIA assay. The schematic was created with BioRender.com.

Article Snippet: Adenylate cyclase-associated protein 1 (CAP1) protein (cat. no. LS-G74326), CAP1 antibodies (cat. no. LS-C381814 and cat. no. LS-C411294), and a human CAP1 ELISA kit (product no. LS-F22718) were purchased from LSBio.

Techniques: Mass Spectrometry, Biomarker Assay, Fluorescence, Generated

Journal: Nature Communications

Article Title: Amplified fluorogenic immunoassay for early diagnosis and monitoring of Alzheimer’s disease from tear fluid

doi: 10.1038/s41467-023-43995-5

Figure Lengend Snippet: a Schematic of the immunocomplex constructed by Ab-MNPs and Ab-PNPs in the presence of the target protein (CAP1) in tear fluid. b Schematic representation of Ab-MNPs labeled with immunogold (Ab-AuNPs) for better visualization of the primary capture antibody bound to the MNPs. c Representative transmission electron microscopy (TEM) image of Ab-MNPs labeled with immunogold (Ab-AuNPs, approximately 10 nm in diameter). Red arrows indicate the localized AuNPs on the surface of Ab-MNPs. The scale bars indicate 100 nm and 50 nm (inset), respectively. d Magnetization curves of MNPs (black) and Ab-MNPs (red) obtained by vibrating sample magnetometer (VSM) at 25 °C. e Schematic of Förster resonance energy transfer (FRET) signal changes of Ab-PNPs induced by treatment with TX-100 surfactant as lysis buffer. f TEM image of Ab-PNPs negatively stained with 3% (w/v) phosphotungstic acid solution (pH 6.81). The scale bar represents 100 nm. g Emission fluorescence spectra of Ab-PNPs before (black circles) and after (red circles) treatment with lysis buffer (excitation at 475 nm). Data represent mean ± s.d. for three independent experiments. The representative images were taken from different samples and repeated at least 50 times independently collection with similar results. Schematics were created with BioRender.com.

Article Snippet: Adenylate cyclase-associated protein 1 (CAP1) protein (cat. no. LS-G74326), CAP1 antibodies (cat. no. LS-C381814 and cat. no. LS-C411294), and a human CAP1 ELISA kit (product no. LS-F22718) were purchased from LSBio.

Techniques: Construct, Labeling, Transmission Assay, Electron Microscopy, Förster Resonance Energy Transfer, Lysis, Staining, Fluorescence

Journal: Nature Communications

Article Title: Amplified fluorogenic immunoassay for early diagnosis and monitoring of Alzheimer’s disease from tear fluid

doi: 10.1038/s41467-023-43995-5

Figure Lengend Snippet: a Detailed process of SNAFIA test using tear fluid. First, (i) tears are combined with Ab-MNPs and Ab-PNPs to form a sandwich immunocomplex with a target protein. (ii) The immunocomplex is subsequently separated from the mixture using a magnet, removing non-target and excess Ab-PNPs. (iii) TX-100 surfactant is added as a lysis buffer to disrupt Ab-PNPs and allow to release of FRET dyes from the Ab-PNPs, amplifying the signal. b Normalized fluorescence intensity (FI) of SNAFIA with increasing concentrations of CAP1 protein spiked in phosphate-buffered saline (PBS, black circles) and artificial tear fluid (ATF, turquoise circles) solution (excitation at 475 nm, emission at 500 nm). The limit of detection (LOD) of SNAFIA in each condition was determined by three-sigma (3σ) and calculated to be 0.282 and 0.236 fM, respectively. c Fluorescence and absorbance signal responses of the SNAFIA (turquoise dots), half-SNAFIA (hSNAFIA, yellowish dots), and ELISA (black dots) tests with different concentrations of CAP1 protein ranging from 10 −2 to 10 8 fM. Normalized FI and absorbance data were fitted to the four-parameter logistic curve (dotted line), and the linear dynamic range of each test is shown by the shaded area. d Selectivity of SNAFIA for various AD biomarker candidates and their mixtures in PBS. The concentrations of CAP1 (turquoise bars), apolipoprotein E (APOE, magenta bars), and acetylcholinesterase (ACHE, orange bars) were varied to 1 pM (1×), 5 pM (5×), and 10 pM (10×). Statistical analysis was performed by multiple comparisons of Brown-Forsythe and Welch one-way analysis of variance tests (**** p < 0.0001). The measurement was performed in triplicate, and all reported values represent mean ± s.d.; n = 3 repeated tests. Schematics were created with BioRender.com.

Article Snippet: Adenylate cyclase-associated protein 1 (CAP1) protein (cat. no. LS-G74326), CAP1 antibodies (cat. no. LS-C381814 and cat. no. LS-C411294), and a human CAP1 ELISA kit (product no. LS-F22718) were purchased from LSBio.

Techniques: Lysis, Fluorescence, Saline, Enzyme-linked Immunosorbent Assay, Biomarker Assay

Journal: Nature Communications

Article Title: Amplified fluorogenic immunoassay for early diagnosis and monitoring of Alzheimer’s disease from tear fluid

doi: 10.1038/s41467-023-43995-5

Figure Lengend Snippet: a Schematic of CAP1 detection by SNAFIA using tear fluid. The SNAFIA test is performed using non-invasively collected tear fluid. The intensity of the fluorescence signals obtained from the SNAFIA test determines the stage of AD progression. b SNAFIA test results showing the normalized FI using human tear fluid from the HC (left), MCI (middle), and AD (right) groups. The pink box labeled patient (patient ID: 16) showed disease progression from MCI to definite AD over two years. c Comparison of normalized FI of the CAP1 biomarker candidate in the human tear fluid of HC, MCI, and AD groups. Each signal analysis of SNAFIA was performed on HC ( n = 14), MCI ( n = 15), and AD ( n = 10) individuals from the verification cohort. Statistical analysis was performed using a one-way analysis of variance (*** p = 0.0003, * p = 0.0251, Kruskal-Wallis test). All measurements were performed in triplicate, and data represent mean ± s.d. d Receiver operating characteristic (ROC) curves of SNAFIA for MCI (yellow dots) or AD (red dots) groups compared with HC individuals. The area under the ROC curve values is shown in the graph. e Correlation between fluorescence signals of SNAFIA using clinical tear samples and the patient’s Mini-Mental State Examination (MMSE) score. The scatter plot shows that the SNAFIA signals and MMSE scores are negatively correlated, with a Pearson correlation coefficient value of −0.8255. Statistical analysis was performed by two-tailed (*** p = 0.0002). Schematics were created with BioRender.com.

Article Snippet: Adenylate cyclase-associated protein 1 (CAP1) protein (cat. no. LS-G74326), CAP1 antibodies (cat. no. LS-C381814 and cat. no. LS-C411294), and a human CAP1 ELISA kit (product no. LS-F22718) were purchased from LSBio.

Techniques: Fluorescence, Labeling, Comparison, Biomarker Assay, Two Tailed Test