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We investigated the relationship between small dense low-density cholesterol (sdLDL-C) and risk of major adverse cardiovascular events (MACE) in patients treated with high- or low-dose statin therapy.This was a prospective case-cohort study within the Randomized Evaluation of Aggressive or Moderate Lipid-Lowering Therapy with Pitavastatin in Coronary Artery Disease (REAL-CAD) study, a randomized trial of high- or low-dose (4 or 1 mg/d pitavastatin, respectively) statin therapy, in patients with stable coronary artery disease (CAD). Serum sdLDL-C was determined using an automated homogenous assay at baseline (randomization after a rule-in period, ＞1 month with 1 mg/d pitavastatin) and 6 months after randomization, in 497 MACE cases, and 1543 participants randomly selected from the REAL-CAD study population.High-dose pitavastatin reduced sdLDL-C by 20% than low-dose pitavastatin (p for interaction ＜0.001). Among patients receiving low-dose pitavastatin, baseline sdLDL-C demonstrated higher MACE risk independent of LDL-C (hazard ratio [95% confidence interval], 4th versus 1st quartile, 1.67 [1.04-2.68]; p for trend=0.034). High-dose (versus low-dose) pitavastatin reduced MACE risk by 46% in patients in the highest baseline sdLDL-C quartile (＞34.3 mg/dL; 0.54 [0.36-0.81]; p=0.003), but increased relative risk by 40% in patients with 1st quartile (≤ 19.5 mg/dL; 1.40 [0.94-2.09]; p=0.099) and did not alter risk in those in 2nd and 3rd quartiles (p for interaction=0.002).These findings associate sdLDL-C and cardiovascular risk, independent of LDL-C, in statin-treated CAD patients. Notably, high-dose statin therapy reduces this risk in those with the highest baseline sdLDL-C.

In this article, we propose a novel microsolid-phase extraction and elution technique, which we called the thin-layer solid-phase extraction-liquid film elution technique. The thin-layer solid-phase extraction phase is an octadecylsilylated sol gel- coated porous silica thin film prepared on the outer wall of a test tube, which has a larger surface area for the extraction of the target compounds compared to a conventional solid-phase microextraction phase. After optimization of the extraction procedure for five types of polycyclic aromatic hydrocarbons, the liquid film elution technique was investigated. Liquid film elution is an elution technique wherein the compounds extracted into the thin-layer solid-phase extraction phase are eluted using a small volume of solvent film formed around the extraction phase. The results show that the elution can be carried out using 150 μL of eluent. Enrichment factors between 20 and 34 were obtained for polycyclic aromatic hydrocarbons containing more than four aromatic rings in 10 mL aliquots of aqueous samples. Finally, recoveries of 85-112% were obtained for polycyclic aromatic hydrocarbons containing more than four aromatic rings from spiked natural water samples using the thin-layer solid-phase extraction-liquid film elution technique.

In this study, we investigated a combination of nonaqueous CE with capillary gel electrophoresis to achieve highly efficient analysis of metal nanoclusters. In the nonaqueous capillary gel electrophoresis (NACGE), PVA and hydroxypropyl methylcellulose were dissolved in DMSO. In addition, to enhance the entanglement of the polymer chains, Li+ ions were also added. By employing the PVA-DMSO-Li+ solution, we studied the effects of the molecular weight, the degree of hydrolysis, and the concentration of the polymers and Li+ on the separation. As a result, good separations of standard mononuclear metal complexes and tetrairon nanoclusters were achieved by NACGE.

An on-line sample preconcentration technique based on transient trapping (tr-trapping) in micellar electrokinetic chromatography (MEKC) was applied for steroid detection with UV (tr-trapping-UV) and electrospray ionization mass spectrometry detection (tr-trapping-ESI-MS). ESI-MS was used to improve the sensitivity in MEKC. The MEKC separation was carried out using volatile ammonium formate as a background solution to facilitate the coupling with ESI-MS. The partial introduction of a sodium dodecyl sulfate (SDS) micellar solution before the introduction of a sample solution to the capillary provided the effective preconcentration of analytes. At the same time, the SDS micelle would not enter the ESI-MS system, so its interference in ESI-MS detection was suppressed under the optimal condition, then five steroids can be separated by the developed method. In tr-trapping-ESI-MS, an acidic condition of pH 3.5 was employed to suppress the electroosmotic flow, which can avoid micellar solution migrating to the MS instrument. The developed method showed that the micellar solution requires a twofold slower time than the sample to migrate along the column, which can prohibit the cause of the problem with the MS instrument and interference signal of SDS in the steroid's detection. The tr-trapping-ESI-MS protocol showed up to 540-fold enhancements of the peak intensity and 50-fold improvement of the limit of detection compared with capillary zone electrophoresis using androsterone as a model sample.

Left-ventricular systolic dysfunction (LVSD) comorbid with atrial fibrillation is reversible, but recovery is limited in a subset of patients. The Selvester QRS (S-QRS) score is an electrocardiogram-based assessment that reportedly reflects myocardial scar/damage. We evaluated the predictability of S-QRS score for the recovery of left-ventricular ejection fraction (LVEF) in persistent AF (PeAF) patients with LVSD undergoing catheter ablation (CA).CA was performed in 51 PeAF patients with reduced LVEF (40%); S-QRS scores were measured after restoration of sinus rhythm. LVEF was re-evaluated at one year after CA; LVEF recovery was related to the S-QRS score.The median [interquartile range] S-QRS score was 1 point [0-2]. LVEF increased from 32% [28-37] at baseline to 56% [49-57] at 1 year after CA. Thirty-seven patients achieved normalization of LVEF (≥50%, Group A); 14 patients did not (Group B). Group A had significantly lower S-QRS scores than Group B (0 point [0-2] vs. 2 points [2-3], p .05). In univariate/multivariate analyses, S-QRS score was an independent predictor of LVEF normalization. In the receiver operating characteristic curve, the cut-off value of S-QRS score was 2 points for prediction of the LVEF normalization (AUC = 0.79). Patients with low S-QRS score (2 points) had greater LVEF improvement than those with high S-QRS score (≥2 points, ΔLVEF: 23% [17-28] vs. 17% [12-24], p .05).S-QRS scoring noninvasively assesses the improvement of LVEF in PeAF patients with LVSD after CA. A high S-QRS score may indicate underlying myocardial scar/damage associated with unknown etiologies for LVSD other than PeAF.

To achieve highly sensitive analysis without labor-intensive experimental procedures in capillary electrophoresis (CE), large-volume sample stacking with an electroosmotic flow pump (LVSEP)-field-amplified sample injection (FASI) was combined with a dynamic coating technique. In this study, poly(vinyl pyrrolidone) (PVP) was employed for the dynamic coating additive. Since a standard fluorescent dye, fluorescein, was well concentrated in a conventional LVSEP, the PVP dynamically-coated capillaries can be also applied to the LVSEP-FASI analysis. In our home-made CE apparatus, however, current breakdown was often caused, especially at a longer electrokinetic injection time due to bubble formation. To avoid the interference of bubble formation, the distance between the tips of the electrode and the capillary in the vertical direction was changed from 0 to 2.5 cm under the magnetic stirring condition. This allowed for a long electrokinetic injection time of up to 20 min, resulting in a sensitive enhancement factor (SEF) of 34900 for fluorescein. The developed method was applied to the chiral analysis of amino acids in CE. As a result, leucine (Leu) was successfully separated in LVSEP-FASI with SEFs of 6420 and 4500 for the D- and L-Leu peaks, respectively.

The prognostic role of D-dimer in different types of heart failure (HF) is poorly understood. We investigated the prognostic value of D-dimer on admission, both independently and in combination with the Get With The Guidelines—Heart Failure (GWTG-HF) risk score and N-terminal pro-B-type natriuretic peptide (NT-proBNP), in patients with preserved left ventricular ejection fraction (LVEF) and acute decompensated HF (HFpEF) or reduced LVEF (HFrEF). Baseline D-dimer levels were measured on admission in 1670 patients (mean age: 75 years) who were hospitalized for worsening HF. Of those patients, 586 (35%) were categorized as HFpEF (LVEF ≥ 50%) and 1084 as HFrEF (LVEF &lt, 50%). During the 12-month follow-up period after admission, 360 patients died. Elevated levels (at least the highest tertile value) of D-dimer, GWTG-HF risk score, and NT-proBNP were all independently associated with mortality in all HFpEF and HFrEF patients (all p &lt, 0.05). Adding D-dimer to a baseline model with a GWTG-HF risk score and NT-proBNP improved the net reclassification and integrated discrimination improvement for mortality greater than the baseline model alone in all populations (all p &lt, 0.001). The number of elevations in D-dimer, GWTG-HF risk score, and NT-proBNP were independently associated with a higher risk of mortality in all study populations (HFpEF and HFrEF patients, all p &lt, 0.001). The combination of D-dimer, which is independently predictive of mortality, with the GWTG-HF risk score and NT-proBNP could improve early prediction of 12-month mortality in patients with acute decompensated HF, regardless of the HF phenotype.

In this study, we investigated a combination of nonaqueous CE with capillary gel electrophoresis to achieve highly efficient analysis of metal nanoclusters. In the nonaqueous capillary gel electrophoresis (NACGE), PVA and hydroxypropyl methylcellulose were dissolved in DMSO. In addition, to enhance the entanglement of the polymer chains, Li

Accessibility and specific surface area are important factors related to the functions and efficiencies of thin films that act as adsorbents, supports for catalysts, and so on. Therefore, controlling the surface morphology or macro porosity has attracted a great deal of attention. In this work, a spontaneous silica film growth process in a sol–gel solution was studied by observing electron microscopy images as well as by using UV-VIS spectroscopy and surface roughness measurements. As a result, a macroporous silica film was formed in a sol–gel solution containing Pluronic P123 as a structure-directing agent. Furthermore, a preparation of macroporous silica films that have different morphologies was attained by using different Pluronic surfactants and inorganic salts. Some films had a micrometer order rod-like structure, and others had a reticular structure composed of a concatenation of short rod-like nanoparticles. Nitrogen adsorption desorption isotherms were measured for silica deposits that formed during film growth in the sol–gel solution to discuss the characteristics of the silica thin films.

A combination of two online sample concentration techniques, large-volume sample stacking with an electroosmotic flow (EOF) pump (LVSEP) and field-amplified sample injection (FASI), was investigated in microchip electrophoresis (MCE) to achieve highly sensitive analysis. By applying reversed-polarity voltages on a cross-channel microchip, anionic analytes injected throughout a microchannel were first concentrated on the basis of LVSEP, followed by the electrokinetic stacking injection of the analytes from a sample reservoir by the FASI mechanism. As well as the voltage application, a pressure was also applied to the sample reservoir in LVSEP-FASI. The applied pressure generated a counter-flow against the EOF to reduce the migration velocity of the stacked analytes, especially around the cross section of the microchannel, which facilitated the FASI concentration. At the hydrodynamic pressure of 15 Pa, 4520-fold sensitivity increase was obtained in the LVSEP-FASI analysis of a standard dye, which was 33-times higher than that obtained with a normal LVSEP. Furthermore, the use of the sharper channel was effective for enhancing the sensitivity, e.g., 29 100-fold sensitivity increase was achieved with the 75-μm wide channel. The developed method was applied to the chiral analysis of amino acids in MCE, resulting in the sensitivity enhancement factor of 2920 for the separated d-leucine.

Modern regenerative medicine and pharmaceutical manufacturing require a large-scale culture of cells. Consequently, cells must be suspension-cultured at high densities, which is challenging because more metabolites, especially ammonia, are produced that inhibit cell proliferation. To address this problem, we tested four zeolites (L-type zeolite, ferrierite, mordenite, and Y-type zeolite) and Prussian blue for their ability to adsorb NH4+ (the predominant form of ammonia in a culture-solution pH) from an aqueous solution and a culture solution. The L-type zeolite and Prussian blue effectively adsorbed NH4+ from aqueous solutions, while the other absorbents were ineffective. Notably, neither adsorbent removed glucose, a representative nutrient, from the aqueous solution. Kinetics and equilibrium studies revealed that both adsorbents (L-type zeolite and Prussian blue) obey pseudo-second-order kinetics and are well fitted by the Langmuir isotherm model. The L-type zeolite also exhibited a high NH4+-adsorption in the culture solution. Further, while the L-type zeolite exhibited minimal cytotoxicity, Prussian blue was cytotoxic. Using the L-type zeolite for adsorption is simple and cost-effective, thus making it a suitable NH4+ adsorbent for culture solutions. Furthermore, reusing an expensive culture solution on a large scale can effectively lower costs of producing cell cultures intended for pharmaceutical manufacturing and regenerative medicine purposes. Thus, it is expected that this method will be used to regenerate culture solutions on a large-scale.

The proliferation of pluripotent stem cells in mass cell culture is hindered by the release of cell metabolites such as lactate and ammonia. In this work, a porous strong-acid cation exchange resin (PK216LH) and Mg–Al layered doubled oxide (LDO) were used to take up NH4+ and lactate, respectively. Initially, PK216LH could take up NH4+ from an aqueous solution. The uptake of NH4+ by PK216LH followed pseudo-second-order kinetics and corresponded to the Langmuir adsorption isotherm, with an apparent activation energy of 28.2 kJ/mol and a maximum adsorption amount of 1.38 mmol/g. The simultaneous use of Mg–Al LDO and PK216LH could not sufficiently take up lactate and NH4+. In contrast, the individual use of Mg–Al LDO and PK216LH in two stages enabled efficient removal of lactate and NH4+.

When cells proliferate in a culture medium, metabolites such as lactic acid gradually accumulate and eventually hinder cell growth. Herein, we considered various layered double hydroxides (LDH) and layered double oxides (LDO) for the selective removal of lactate in aqueous solution and a culture medium by anion exchange. NO3•Mg-Al LDH showed high lactate adsorption, low adsorption of glucose (a nutrient in culture medium), appropriate pH stability, and low cytotoxicity within a limited contact time. In contrast, NO3•Ni-Al LDH, NO3•Ca-Al LDH, Mg-Al LDO, and Cl•Mg-Al LDH had a lower performance than NO3•Mg-Al LDH. To further improve the performance, the anion of 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES, a common pH buffer in culture media) was tested as the intercalated anion in Mg-Al LDH. HEPES•Mg-Al LDH (Mg/Al = 2) was found to be the most promising adsorbent for culture medium regeneration because its adsorption behavior was similar to that of NO3•Mg-Al LDH, while it had a much lower cytotoxicity. The traditional methods for regenerating a culture medium by removing accumulated lactate to increase the cell density in large-scale cell culture and lower the costs, including electrolysis, cell modification, and modification of the culture medium’s composition, are complicated to operate and expensive. The adsorption method using HEPES•Mg-Al LDH presented in this study, instead, can be easily operated and has low costs. Thus, it is expected to be applied in actual cases for regenerating culture media.

BACKGROUND A modestly elevated circulating D-dimer level may be relevant to coronary artery disease (CAD), but its prognostic value, both independently and in combination with estimated glomerular filtration rate (eGFR), for long-term death has not been fully evaluated in stable CAD patients.Methods and Results:Baseline plasma D-dimer levels and eGFR were measured in 1,341 outpatients (mean age: 65 years) with prior myocardial infarction (MI), coronary revascularization, and/or angiographic evidence of a significant stenosis (>50%) for at least one of the major coronary arteries. Among these patients, 43% had prior MI, 47% had prior coronary revascularization, 41% had multivessel CAD, 14% had paroxysmal or persistent atrial fibrillation, 32% had diabetes, and 32% had chronic kidney disease (eGFR

To achieve an on-line coupling of the sample preconcentration by a large-volume sample stacking with an electroosmotic flow pump (LVSEP) with microchip gel electrophoresis (MCGE), a sample solution, a background solution for LVSEP and a sieving solution for MCGE were loaded in a T-form channel and three reservoirs on PDMS microchips. By utilizing the difference in the flow resistance of the two channels, a low-viscosity sample and a viscous polymer solution were easily introduced into the LVSEP and MCGE channels, respectively. Fluorescence imaging of the sequential LVSEP-MCGE processes clearly demonstrated that a faster stacking of anionic fluorescein and successive introduction into the MCGE channel can be carried out on the T-channel chip. To evaluate the preconcentration performance, a conventional MCZE analysis of fluorescein on the cross-channel chip was compared with LVSEP-MCGE on the short T-channel chip, and as a result that the value of sensitive enhancement factor (SEF) was estimated to be 370. The repeatability of the peak height was good with the RSD value of 3.2%, indicating the robustness of the enrichment performance. In the successive LVSEP-MCGE analysis of φX174/HaeIII digest, the DNA fragments were well enriched to a sharp peak in the LVSEP channel, and they were separated in the MCGE channel, whose electropherogram was well-resembled with that in the conventional MCGE. The values of SEF for the DNA fragments were calculated to be ranging from 74 to 108. Thus, the successive LVSEP-MCGE analysis was effective for both preconcentrating and separating DNA fragments.

Lactate is an important metabolite generated during the culture of pluripotent stem cells and is known to inhibit the cell growth. With the increase in the prominence of pluripotent stem cell applications, reliable methods are required to remove metabolites such as lactate for enabling the high-density culture of cells. Herein, we report an investigation of layered double hydroxides (LDHs) and layered double oxides (LDOs) as anion-adsorbents for lactate removal to regenerate the cell culture medium. The lactate absorption ability of Cu-Al and Mg-Al LDH and Cu-Al and Mg-Al LDO adsorbents was evaluated. The influence of various process parameters, including temperature and pH, on the lactate adsorption, were investigated. The kinetics of the adsorption process and equilibrium dynamics were studied, and the maximum adsorption amounts were determined. The Cu-Al LDH, Cu-Al LDO, Mg-Al LDH, and Mg-Al LDO exhibited a lactate adsorption that followed a pseudo-second-order kinetic model, and their apparent activation energies for adsorption were 32.0, 47.4, 95.7, and 61.7 kJ/mol, respectively; these values indicated that chemical adsorption was the underlying mechanism of adsorption. The Langmuir and Freundlich models were analyzed and revealed the nature of the anion-exchange process. This study is expected to promote the high-density culture of cells by ensuring the regeneration of the media by the efficient removal of the undesired lactate.

In this study, we found that the polarity switching was effective to enrich and separate fluorescent analytes which have weakly-dissociated groups in a floating platinum electrode (width, 50 µm; thickness, 2.5 µm)-integrated straight-channel in microchip electrophoresis (MCE). In the straight channel filled with an Alexa Flour 488 (AF488) solution, a sharp peak was observed after the polarity inversion with a 530-fold enhancement of the sensitivity relative to the conventional MCE analysis. By using a fluorescent pH indicator, we verified that a sharp high-pH zone was generated nearby the floating electrode and moved toward the anode with maintaining the high pH, which induced the sample enrichment like a dynamic pH junction mechanism. In the floating electrode-embedded channel, the mixture of AF488-labeled proteins was also well concentrated and separated within 100 s.

Electrophoretic on-line sample preconcentration techniques in microfluidic channels improve the sensitivity prior to the separation. Among various techniques, the most important field-amplified sample stacking and sweeping on cross-channel microchips are demonstrated. As a novel microfluidic preconcentration approach, a large-volume sample stacking with electroosmotic flow pump (LVSEP) on straight-channel chips is also presented, which can omit a complicated voltage program for sample injection processes. In this chapter, we describe how to prepare and how to run these on-line sample preconcentration methods in microchip electrophoresis.

Electrophoretic on-line sample preconcentration techniques in microfluidic channels improve the sensitivity prior to the separation. Among various techniques, the most important field-amplified sample stacking and sweeping on cross-channel microchips are demonstrated. As a novel microfluidic preconcentration approach, a large-volume sample stacking with electroosmotic flow pump (LVSEP) on straight-channel chips is also presented, which can omit a complicated voltage program for sample injection processes. In this chapter, we describe how to prepare and how to run these on-line sample preconcentration methods in microchip electrophoresis.

Background The early prediction of acute kidney injury (AKI) can facilitate timely intervention and prevent complications. We aimed to understand the predictive value of urinary liver-type fatty-acid binding protein (L-FABP) levels on admission to medical (non-surgical) cardiac intensive care units (CICUs) for AKI, both independently and in combination with serum N-terminal pro-B-type natriuretic peptide (NT-proBNP) levels. Methods We prospectively investigated the predictive value of L-FABP and NT-proBNP for AKI in a large, heterogeneous cohort of patients treated in medical CICUs. Baseline urinary L-FABP and serum NT-proBNP were measured on admission. AKI was diagnosed according to the Kidney Disease: Improving Global Outcomes criteria. We studied 1273 patients (mean age, 68 years), among whom 46% had acute coronary syndromes, 38% had acute decompensated heart failure, 5% had arrhythmia, 3% had pulmonary hypertension, 2% had acute aortic syndrome, 2% had infective endocarditis, and 1% had Takotsubo cardiomyopathy. Results Urinary L-FABP levels correlated with serum NT-proBNP levels (r = 0.17, p

A simple and rapid vacuum-drying modification method was applied to several neutral and charged polymers to obtain coating layers for controlling electroosmotic flow (EOF) and suppressing sample adsorption on poly(dimethyl siloxane) (PDMS)-glass hybrid microchips. In the vacuum-dried poly(vinylpyrrolidone) coating, the electroosmotic mobility (μeo) was suppressed from +2.1 to +0.88 × 10(-4) cm(2)/V·s, and the relative standard deviation (RSD) of μeo was improved from 10.2 to 2.5% relative to the bare microchannel. Among several neutral polymers, poly(vinylalcohol) (PVA) and poly(dimethylacrylamide) coatings gave more suppressed and repeatable EOF with RSDs of less than 2.3%. The vacuum-drying method was also applicable to polyanions and polycations to provide accelerated and inversed EOF, respectively, with acceptable RSDs of less than 4.9%. In the microchip electrophoresis (MCE) analysis of bovine serum albumin (BSA) in the vacuum-dried and thermally-treated PVA coating channel, an almost symmetric peak of BSA was obtained, while in the native microchannel a significantly skewed peak was observed. The results demonstrated that the vacuum-dried polymer coatings were effective to control the EOF, and reduced the surface adsorption of proteins in MCE.

BACKGROUND We prospectively investigated the prognostic value of the combined use of cardiac troponin T (TnT), B-type natriuretic peptide (BNP), and high-sensitivity C-reactive protein (CRP) for long-term mortality in hemodialysis (HD) patients. METHODS AND RESULTS: Baseline measurements of TnT, BNP, and CRP were performed in 516 patients on chronic HD. Patients were followed up for 10 years. Using the Cox multivariate model with these 3 biomarkers as variables categorized into tertiles for mortality, a simplified score was obtained by underscoring individual biomarkers based on the adjusted hazard ratio (HR). The multimarker score was defined as the sum of these points. TnT, BNP, and CRP levels were individually independent predictors for mortality (P

A combination of two online sample concentration techniques, large-volume sample stacking with an electroosmotic flow (EOF) pump (LVSEP) and field-amplified sample injection (FASI), was investigated in microchip electrophoresis (MCE) to achieve highly sensitive analysis. By applying reversed-polarity voltages on a cross-channel microchip, anionic analytes injected throughout a microchannel were first concentrated on the basis of LVSEP, followed by the electrokinetic stacking injection of the analytes from a sample reservoir by the FASI mechanism. As well as the voltage application, a pressure was also applied to the sample reservoir in LVSEP-FASI. The applied pressure generated a counter-flow against the EOF to reduce the migration velocity of the stacked analytes, especially around the cross section of the microchannel, which facilitated the FASI concentration. At the hydrodynamic pressure of 15 Pa, 4520-fold sensitivity increase was obtained in the LVSEP-FASI analysis of a standard dye, which was 33-times higher than that obtained with a normal LVSEP. Furthermore, the use of the sharper channel was effective for enhancing the sensitivity, e.g., 29 100-fold sensitivity increase was achieved with the 75-μm wide channel. The developed method was applied to the chiral analysis of amino acids in MCE, resulting in the sensitivity enhancement factor of 2920 for the separated d-leucine.

Urinary liver-type fatty acid-binding protein (L-FABP) reflects the degree of stress in proximal tubules of the kidney. We examined the level of L-FABP in type-2 diabetes mellitus (T2DM) patients with chronic kidney disease (CKD) stage G1 and G2, and its relationship with cardiac markers and electrocardiographic (ECG) abnormalities. T2DM patients whose estimated glomerular filtration rate (eGFR) was ≥60 mL/min/1.73 m(2) were recruited [n = 276 (165 males), mean age 64 years]. The median level of urinary L-FABP was 6.6 μg/gCr. Urinary L-FABP showed significant correlation with urinary albumin-to-creatinine ratio (ACR) (r = 0.51, p0.0001). Median (25th-75th percentile) eGFR was 82 (72-95) mL/min/1.73 m2. We divided patients into four subgroups (group 1, L-FABP ≤8.4 μg/gCr and ACR ≤30 mg/gCr; group 2, L-FABP ≤8.4 μg/gCr and ACR30 mg/gCr; group 3, L-FABP8.4 μg/gCr and ACR ≤30 mg/gCr; group 4, L-FABP8.4 μg/gCr and ACR30 mg/gCr). Compared with group 1, group 4 was significantly higher in systolic blood pressure, and eGFR using standardized serum cystatin C, high-sensitivity troponin T, and N-terminal pro-brain natriuretic peptide (NT-proBNP). Group 4 had significantly higher level of NT-proBNP than group 3. Groups 2, 3 and 4 showed more ECG abnormalities than group 1. These findings suggest that simultaneous measurement of urinary L-FABP and ACR should be useful to assess cardiovascular damage reflecting on the elevation of cardiac markers and ECG abnormalities in T2DM with CKD G1 and G2.

Chronic kidney disease (CKD) is associated with an increased risk of cardiovascular (CV) events. Recently, elevated neutrophil gelatinase-associated lipocalin (NGAL) levels have been reported in patients with heart failure, coronary heart disease, or stroke. Our aim was to assess urinary NGAL as a predictor of CV events in patients with CKD. This was a prospective observational cohort study of 404 patients with predialysis CKD. CV events were defined as CV death, acute coronary syndrome, hospitalization for worsening heart failure, stroke and dissection of aorta. During a mean follow-up period of 33 months, 77 CV events (19.1 %) occurred. After adjustment for gender, age, diabetes, previous cardiovascular disease, urinary albumin/creatinine ratio (UACR), estimated glomerular filtration rate, hemoglobin, and high-sensitivity C-reactive protein, patients with the other quartiles of urinary NGAL had significantly higher risk of CV events compared with patients with the lowest quartile (hazard ratio (HR) 2.81, 95 % confidence interval (CI) 1.01–7.81, P = 0.047 for Q2, HR 3.31, 95 % CI 1.22–9.00, P = 0.019 for Q3, and HR 3.27, 95 % CI 1.15–9.29, P = 0.026 for Q4). Regarding the combination of urinary NGAL with UACR, we also stratified patients into four groups according to whether the level of each marker was above or below the median (61.8 μg per gram creatinine (gCr) for NGAL and 351.1 mg/gCr for UACR). Four-year CV event-free survival rates were 89.2, 79.6, 71.8, and 51.5 % in order for the four respective groups (P < 0.0001). Elevated urinary NGAL was able to predict future CV events in CKD patients, and had incremental predictive value with elevated UACR.

Human mesenchymal stem cells (hMSCs) that can differentiate into chondrocytes are a potential autologous cell source for repair of damaged tissue. Current methods usually induce the formation of all three chondrocyte phenotypes, hyaline, fibrous, and elastic, without the ability to selectively induce only one of them. By controlling the size of hMSC cell clusters, it may be possible to direct differentiation more uniformly toward hyaline chondrocytes. We designed new cell culture platforms containing microwells of different diameters. The platforms and wells were composed of a zirconia ceramics substratum. hMSCs briefly adhered to the substratum before releasing and entering the microwells. The physical restraints imposed by the microwells enabled hMSC clusters to homogenously differentiate into hyaline chondrocyte-like cells. Chondrogenic aggregates in microwells expressed the hyaline chondrocyte-specific genes Col II, aggrecan (ACAN), and cartilage oligomeric protein (COMP). The cultures also produced hyaline chondrocyte-specific matrix proteins Col II and ACAN homogenously throughout the aggregates. In contrast, chondrogenesis in pellet cultures was heterogeneous with the expression of nonhyaline chondrocyte genes CD105, Col X, and Col I. In these pellet cultures, hyaline and nonhyaline chondrocyte-specific matrix proteins were distributed heterogeneously. Thus, this novel ceramic microwell substratum technology efficiently directed the differentiation of hyaline chondrocyte-like cells from hMSCs. These results indicate that there is a close relationship between hMSC cluster size regulation in the microwells and differentiation tendency. This microwell culture differentiation method will provide a valuable experimental system for both experimental and potential clinical studies.

Background Several epidemiologic studies have reported an inverse association between serum levels of carotenoids and cardiovascular disease risk. However, no studies have reported an association between serum carotenoids and N-terminal pro-brain-type natriuretic peptide (NT-proBNP) in the general population. Methods In this cross-sectional study, we investigated whether serum carotenoids were associated with serum NT-proBNP in 1056 Japanese subjects (390 men, 666 women) who attended a health examination. Serum levels of carotenoids were separately determined by high-performance liquid chromatography. Serum NT-proBNP level was measured by electrochemiluminescence immunoassay. Results Serum NT-proBNP was elevated (≥55 pg/ml) in 31.8% of men and 48.2% of women. Multivariate logistic regression analyses adjusted for confounding factors showed a significant association between the highest quartile of serum α-carotene and elevated NT-proBNP in men (odds ratio [OR] = 0.40, 95% CI = 0.19-0.82, P for trend = 0.005) and women (OR = 0.62, 95% CI = 0.39-0.99, P for trend = 0.047). In women, moreover, elevated serum NT-proBNP was significantly associated with serum canthaxanthin (OR = 0.57, 95% CI = 0.36-0.90 for highest quartile, P for trend = 0.026) and β-cryptoxanthin (OR = 0.53, 95% CI = 0.32-0.85 for highest quartile, P for trend = 0.026), after adjusting for potential confounders. Conclusions Higher levels of serum carotenoids were associated with lower risk of elevated serum NT-proBNP levels after adjusting for possible confounders, which suggests that a diet rich in carotenoids could help prevent cardiac overload in the Japanese population.

The effects of a low-conductivity (low-C) zone of an on-line sample preconcentration based on field-amplified sample stacking (FASS) under an acidic condition was investigated in microchip micellar electrokinetic chromatography (MCMEKC). By employing originally fabricated microchips with T- and cross-channel injectors on a single device, partial injection of the low-C zone and large-volume injection of a sample solution were demonstrated. As a typical result, the observed peak intensities of the fluorescent dyes were 1.5 - 2.0-fold enhanced by partial injection of the low-conductivity zone, as compared to those in FASS-MCMEKC without injecting the low-C zone; also, 30 - 40-fold enhancements of the detectability were achieved as compared to the conventional MCMEKC analyses. The resolutions of the hydrophobic analytes were also improved by introducing the low-C zone. The difference in the retention factors of model analytes in the high-C background and low-C zones was also estimated in order to clarify the effect of the low-C zone on the resolution and detectability in FASS-MCMEKC.

The preparation of cinchonidine (CCND) nanocrystals and their immobilization onto the inner surface of a fused silica capillary was investigated for use in the enantioseparation by capillary electrochromatography. The CCND nanocrystals were prepared by an emulsion method that utilizes crystal growth in an oil-in-water emulsion without a stabilizer. As a result, white-colored aqueous dispersions of organic nanoparticles were obtained. SEM images showed that the prepared CCND nanocrystals were from 300 to 700 nm in size. When the obtained dispersion was introduced into a poly(diallyldimethylammonium chloride) (PDDAC) coated capillary, the nanocrystals were strongly adsorbed onto the inner surface due to an electrostatic interaction between cationic PDDAC and the negatively charged organic nanocrystals. In CCND nanocrystals coated capillary, CEC enantioseparations of racemic amino acids were successfully achieved. The reproducibilities of the detection times were acceptable with a relative standard deviation of less than 7%, indicating that stable nanocrystal coating could be obtained by our proposed method.

Whether trough-phase rivaroxaban concentrations provide sufficient anticoagulation needs more study. We evaluated levels of coagulation activation markers in the trough concentration phase in nonvalvular atrial fibrillation (NVAF) patients, and the correlation between these markers and rivaroxaban concentration. Fifty-five Japanese NVAF patients received 24-week rivaroxaban treatment of either 15 or 10 mg once-daily in the morning. Of these, 26 patients had no history of anticoagulant therapy (naive group) and 29 had switched from warfarin (warfarin group). D-dimer and prothrombin fragment 1 + 2 (F1 + 2) levels, and protein C activities were measured at 0 (baseline), 12 and 24 weeks of rivaroxaban treatment just before the patient’s regular dosing time (trough phase). For 49 patients, D-dimer, F1 + 2, and rivaroxaban concentrations were also measured twice between 28 and 32 weeks of rivaroxaban treatment at non-trough times to achieve a range of drug concentrations for correlation analysis. For the naive group, D-dimer and F1 + 2 levels were significantly reduced (p

Background. Our aim was to assess plasma neutrophil gelatinase-associated lipocalin (NGAL) as a predictor of cardiovascular (CV) events in patients with chronic kidney disease (CKD) and no history of CV events.Methods. This was a prospective observational cohort study of 252 patients with predialysis CKD. CV events were defined as CV death, acute coronary syndrome, and hospitalization for worsening heart failure, stroke, and aortic dissection.Results. During a median follow-up period of 63 months, 36 CV events occurred. On Cox stepwise multivariate analysis, plasma NGAL and B-type natriuretic peptide (BNP) were significant predictors of CV events. Kaplan-Meier incidence rates of CV event-free survival at 5 years were 96.6%, 92.9%, 85.9%, and 61.3%, respectively, among quartiles of plasma NGAL (P<0.0001). The C-index for the receiver-operating characteristic curves for CV events was greater when plasma NGAL was added to an established risk model (0.801, 95% CI 0.717–0.885), compared to the model without plasma NGAL (0.746, 95% CI 0.653–0.840,P=0.021).Conclusion. Elevated plasma NGAL could predict future CV events in CKD patients with no history of CV events and add incremental value to the established risk model.

To prepare mesoporous silica thin-films by a simple and environmentally friendly pathway, spontaneous formation of silica thin-films in a sol–gel solution without surfactants was studied. The silica thin-films were prepared by immersing a glass substrate, such as a cylindrical test-tube, into a sol–gel solution containing tetraethoxysilane, water, acetic acid, 2-propanol, and triethylene glycol as a pore-forming agent. The resulting thin-films were transparent. The presence of mesopores was confirmed by field-emission scanning electron microscopy, atomic-force microscopy and nitrogen adsorption–desorption isotherms. The Barrett–Joyner–Halenda plot from the adsorption branch of the isotherms indicated that the pore diameter was 2.6 nm. To illustrate the potential use of the film as an adsorbent, the extraction of polycyclic aromatic hydrocarbons was conducted using the chemically modified silica thin-film coated on a test-tube. The composition of the sol–gel solution was optimized using experimental design, i.e., Doehlert design, and recovery of polycyclic aromatic hydrocarbons.

An on-line sample preconcentration based on transient trapping was applied to cyclodextrin electrokinetic chromatography (CDEKC) to realize a highly sensitive chiral analysis in capillary electrophoresis. The partial introduction of a micellar solution before the electrokinetic injection of a sample solution provided the effective preconcentration and enantioseparation of chiral compounds, resulting in the up to 240-fold enhancements of peak intensity and 100-fold improvements of limit of detection of (R),(S)-1-aminoindan as the model analyte. The demonstrated method could be applied to the other pharmaceutical compounds, which allowed five chiral analytes to be resolved with 40- to 160-fold sensitivity enhancements at once. The enantioseparation efficiency of the proposed method was slightly lower as compared to the conventional CDEKC, while the acceptable baseline separations of enantiomers were obtained in transient trapping-CDEKC relative to the undesirable resolution in the CDEKC with other preconcentration techniques. Additionally, it was clarified that transient trapping-CDEKC was also applicable to the analysis of enantiomeric excess, providing the sensitive detection of 43 ppb of (R)-chlorpheniramine in 5 ppm (S)-chlorpheniramine solution commercially available.

To realize the high-performance and simple-operation analysis of cationic compounds in capillary electrophoresis, we investigated large-volume sample stacking with an electroosmotic flow pump (LVSEP) using capillaries with hydrophilic and weakly cationic inner surface. Three capillary modification methods were employed: thermally passivated physical coating with polymer mixture of poly(vinyl alcohol) and poly(allylamine); covalent modification with random copolymer of acryl amide and 3-(methacryloylamino)propyltrimethylammonium chloride; easily preparable physical coating with dimethyldioctadecylammonium bromide and polyoxyethylene stearate. In these capillaries, the electroosmotic flow (EOF) was well suppressed in the high ionic strength (I) electrolyte under the acidic and basic pH, whereas the EOF was enhanced in the low I electrolyte, indicating a suitable EOF property for the rapid LVSEP and following separation. In the LVSEP-capillary zone electrophoresis (CZE) analyses of benzylamine and 1-naphthylethylamine, up to 550-fold sensitivity increases were successfully obtained in the three capillaries without significantly reducing the repeatability and resolution. LVSEP-cyclodextrin-modified CZE of chlorpheniramine and brompheniramine was also carried out, resulting in up to 380-fold sensitivity enhancement with keeping the baseline separation for the enantiomers. Finally, we performed the LVSEP-CZE analysis of basic proteins, where up to 100-fold sensitivity increases were achieved, but a peak broadening was observed due to the sample adsorption in the low I sample matrix.

ObjectivesThe clinical benefit of ezetimibe, an intestinal cholesterol transporter inhibitor, for treatment of postprandial hyperlipidemia was assessed in subjects who ingested a high-fat and high-glucose test meal to mimic westernized diet.MethodsWe enrolled 20 male volunteers who had at least one of the following: waist circumference ≥ 85cm, body mass index ≥ 25kg/m2, or triglycerides (TG) from 150 to 400mg/dL. After 4 weeks of treatment with ezetimibe (10mg/day), the subjects ingested a high-fat and high-glucose meal. Then changes in serum lipid and glucose levels were monitored after 0, 2, 4, and 6h, and the area under the curve (AUC) was calculated for the change in each parameter.Results and conclusionAt 4 and 6h postprandially, TG levels were decreased (p

Serum troponin T levels using a highly sensitive assay (hsTnT) in patients with chronic kidney disease (CKD) not on dialysis have not been examined. The aim of this prospective cohort study was to investigate the association of hsTnT with cardiac events in ambulatory CKD patients not on dialysis. The serum hsTnT level was measured in 442 ambulatory CKD patients not on dialysis whose estimated glomerular filtration rate was

To improve the sensitivity in chiral analysis by capillary electrophoresis without loss of optical resolution, application of large-volume sample stacking with an electroosmotic flow pump (LVSEP) was investigated. Effects of the addition of cyclodextrin (CD) into a running solution on the LVSEP preconcentration was theoretically studied, where the preconcentration efficiency and effective separation length would be slightly increased if the effective electrophoretic velocity (v(ep, eff, BGS)) of the analytes was decreased by interacting with CD. In LVSEP-CD-modified capillary zone electrophoresis (CDCZE) and LVSEP-CD electrokinetic chromatography with reduced v(ep, eff, BGS), up to 1000-fold sensitivity increases were achieved with almost no loss of resolution. In LVSEP-CD-modified micellar electrokinetic chromatography of amino acids with increased v(ep, eff, BGS), a 1300-fold sensitivity increase was achieved without much loss of resolution, indicating the versatile applicability of LVSEP to many separation modes. An enantio-excess (EE) assay was also carried out in LVSEP-CDCZE, resulting in successful analyses of up to 99.6% EE. Finally, we analyzed ibuprofen in urine by desalting with a C(18) solid-phase extraction column. As a typical result, 250ppb ibuprofen was well concentrated and optically resolved with 84.0-86.6% recovery in LVSEP-CDCZE, indicating the applicability of LVSEP to real samples containing a large amount of unnecessary background salts.