Dual Stimuli-Responsive Phenylboronic Acid-Containing Framboidal Nanoparticles by One-Step Aqueous Dispersion Polymerization

Phenylboronic acid-containing nanomaterials have found applications in various fields including biomedical engineering due to their unique stimuli-responsive characteristics. Contrary to the many reports on spherical nanoparticles, we are interested in nanostructures with different morphology which could potentially exhibit additional morphology-related effects. Here, phenylboronic acid-containing nanoparticles (PBANPs) in the size range of 80–250 nm in diameter were synthesized via aqueous dispersion polymerization of <i>N</i>-acryloyl-3-aminophenyl­boronic acid (PBAAM) using methoxy poly­(ethylene glycol) acrylamide (PEGAM) as a polymerizable dispersant and <i>N</i>,<i>N</i>′-methylenebis­(acrylamide) (MBAM) as a cross-linker. Microscopic analysis revealed that PBANPs were clusters, composed of smaller primary nanoparticles of ∼20 nm in diameter, possessing a framboidal morphology. The size of the PBANPs was significantly affected by the concentrations of PBAAM and PEGAM. Furthermore, PBANPs showed reversible swelling behavior in response to the changes in pH and fructose concentration. PBANPs could be used for fructose detection by the PBA-Alizarin Red S displacement assay. The unique framboidal morphology together with the characteristic properties of phenylboronic acid groups may be useful in biosensing applications

Dual Stimuli-Responsive Phenylboronic Acid-Containing Framboidal Nanoparticles by One-Step Aqueous Dispersion Polymerization

Phenylboronic acid-containing nanomaterials have found applications in various fields including biomedical engineering due to their unique stimuli-responsive characteristics. Contrary to the many reports on spherical nanoparticles, we are interested in nanostructures with different morphology which could potentially exhibit additional morphology-related effects. Here, phenylboronic acid-containing nanoparticles (PBANPs) in the size range of 80–250 nm in diameter were synthesized via aqueous dispersion polymerization of <i>N</i>-acryloyl-3-aminophenyl­boronic acid (PBAAM) using methoxy poly­(ethylene glycol) acrylamide (PEGAM) as a polymerizable dispersant and <i>N</i>,<i>N</i>′-methylenebis­(acrylamide) (MBAM) as a cross-linker. Microscopic analysis revealed that PBANPs were clusters, composed of smaller primary nanoparticles of ∼20 nm in diameter, possessing a framboidal morphology. The size of the PBANPs was significantly affected by the concentrations of PBAAM and PEGAM. Furthermore, PBANPs showed reversible swelling behavior in response to the changes in pH and fructose concentration. PBANPs could be used for fructose detection by the PBA-Alizarin Red S displacement assay. The unique framboidal morphology together with the characteristic properties of phenylboronic acid groups may be useful in biosensing applications

A Development of Nucleic Chromatin Measurements as a New Prognostic Marker for Severe Chronic Heart Failure

<div><p>Background</p><p>Accurate prediction of both mortality and morbidity is of significant importance, but it is challenging in patients with severe heart failure. It is especially difficult to detect the optimal time for implanting mechanical circulatory support devices in such patients. We aimed to analyze the morphometric ultrastructure of nuclear chromatin in cardiomyocytes by developing an original clinical histopathological method. Using this method, we developed a biomarker to predict poor outcome in patients with dilated cardiomyopathy (DCM).</p><p>Methods and Results</p><p>As a part of their diagnostic evaluation, 171 patients underwent endomyocardial biopsy (EMB). Of these, 63 patients diagnosed with DCM were included in this study. We used electron microscopic imaging of cardiomyocyte nuclei and an automated image analysis software program to assess whether it was possible to detect discontinuity of the nuclear periphery. Twelve months after EMB, all patients with a discontinuous nuclear periphery (Group A, n = 11) died from heart failure or underwent left ventricular assist device (VAD) implantation. In contrast, in patients with a continuous nuclear periphery (Group N, n = 52) only 7 patients (13%) underwent VAD implantation and there were no deaths (p<0.01). We then evaluated chromatin particle density (Nuc-CS) and chromatin thickness in the nuclear periphery (Per-CS) in Group N patients; these new parameters were able to identify patients with poor prognosis.</p><p>Conclusions</p><p>We developed novel morphometric methods based on cardiomyocyte nuclear chromatin that may provide pivotal information for early prediction of poor prognosis in patients with DCM.</p></div

Distribution of Chromatin Score Values (Nuc-CS and Per-CS) and Cardiac Events.

<p>(A) Relationship between Nuc-CS and cardiac events. A white circle represents a patient who did not experience a cardiac event and a dark circle represents a patient who experienced a cardiac event. (B) Relationship between Per-CS and cardiac events. A white circle indicates a patient who did not experience a cardiac event and a dark circle represents a patient who experienced a cardiac event. (C) Receiver operating characteristic (ROC) curve analysis for Nuc-CS in predicting ventricular assist device (VAD) implantation 1 year after endomyocardial biopsy (EMB). (D) ROC curve analysis for Per-CS predicting VAD implantation 1 year after EMB. (E) ROC curve analysis for BNP (black), LVEDD (light blue), LVEF (dark blue), and %Fib (gray) in predicting VAD implantation 1 year after EMB. AUC, area under the curve; BNP, brain natriuretic peptide; LVEDD, left ventricular end-diastolic diameter; LVEF, left ventricular ejection fraction; %Fib, area of fibrosis within a specimen.</p

Algorithm for DCM Therapy Based on Myocardial Biopsy Parameters.

<p>The first step is morphometric analysis of cardiomyocyte nuclei. When chromatin in the nuclear periphery is discontinuous and there is a homogeneous aggregation of particles in the nucleoplasm, the patient is classified as Group A, which is associated with poor prognosis. When a patient does not have these characteristics, a second step, which involves additional quantitative analysis, is added. Based on the chromatin scores Nuc-CS and Per-CS, a treatment plan can be determined.</p

Clinical Outcomes of Groups A and N.

<p>Clinical Outcomes of Groups A and N.</p

Representative Nuclei of 6 Patients from Each Group.

<p>Representative images are shown for each group. Six patients from Group A (A)–(F). Six patients from Group N: (G)–(L). Scale bar: 2 μm.</p

Relationship between the Nucleoplasmic Chromatin Score (Nuc-CS), Perinuclear Chromatin Score (Per-CS), and Clinical Parameters.

<p>Scatter plots of clinical parameters are shown. % Fib: area of fibrosis within the specimen, LVEF: left ventricular ejection fraction, LVEDD: left ventricular end-diastolic diameter, BNP: brain natriuretic peptide. p: correlation coefficient.</p

Classification of Groups A and N Based on Quantitative Analysis.

<p>(A) Representative original electron microscopic images before digital analysis are shown (10,000× magnification). The perinuclear condensed chromatin was isolated automatically using image analysis software. The area of perinuclear condensed chromatin, which is defined as the area with a grayscale value between 146 and 256, is outlined in red. Black arrows indicate areas of discontinuous nuclear periphery (discontinuous signals of perinuclear condensed chromatin by automatic detection). (B) Two types of nuclei, Group A and Group N, are shown. Representative conventional electron microscopic images (80 kV). Group A: These nuclei have discontinuous perinuclear condensed chromatin with irregular margins and unclear borders. There are homogeneous aggregations of particles in the nucleoplasm. Group N: These nuclei have continuous margins and clear borders. The granular particles in the nucleoplasm occasionally accumulate in spots, but do not aggregate closely. Scale bar: 1 μm. (C) High-power conventional electron microscopic (100 kV) images of chromatin structure in Patients 1 and 2, respectively. In Patient 1, the nucleus has an aggregated structure composed of poorly defined particles. In Patient 2, the nucleus has relatively sparse accumulations of particles less than 30 nm in size. Scale bar: 200 nm.</p