Extremely Simple and Rapid HPLC Analysis of Tocilizumab in Human Serum with Selective Precipitation Using Alkylamine.

An assay using HPLC with fluorescence (FL) detection method for monitoring native FL of tocilizumab (TCZ) in human serum combined with extremely simple and rapid pretreatment without any antigen-antibody reaction was developed. Good separation of TCZ was achieved within 13 min on a Presto FF-C18 column (100 × 4.6 mm i.d., 2 µm). Simple pretreatment with acetonitrile containing primary and secondary alkylamines having longer than C3 in the alkyl chain removed immunoglobulin G subclass 1 and TCZ could be recovered selectively. The spiked calibration curve of TCZ in human serum showed good linearity in the range of 40-1000 µg/mL (r > 0.997). The lower limit of quantitation (S/N = 10) of the TCZ was 19.7 µg/mL. The accuracy was within 103.5-114.9%, and the intra- and inter-day precisions as relative standard deviations were less than 5.3 and 7.8% (n = 5), respectively. The recovery of TCZ was 42.2 ± 3.4% (n = 3). The TCZ in pretreated sample was confirmed to be stable for 6 h (>95%) at room temperature and 24 h (>95%) at 4 °C. The proposed method is considered extremely superior to the previous methods in terms of time requirement for analysis. Therefore, the developed method may be more useful than conventional methods in urgent situations, such as confirming therapeutic efficacy of cytokine-release syndrome by 2019 coronavirus disease.

Safety and efficacy of uninterrupted treatment with edoxaban or warfarin during the peri-procedural period of catheter ablation for atrial fibrillation.

BACKGROUND: The real-world safety and efficacy of uninterrupted anticoagulation treatment with edoxaban (EDX) or warfarin (WFR) during the peri-procedural period of catheter ablation (CA) for atrial fibrillation (AF) are yet to be investigated. METHODS: We conducted a two-center experience, observational study to retrospectively investigate consecutive patients who underwent CA for AF and received EDX or WFR. We examined the incidence of thromboembolic and bleeding complications during the peri-procedural period. RESULTS: The EDX and WFR groups included 153 and 103 patients, respectively (total: 256 patients). Demise or thromboembolic events did not occur in either of the groups. The incidence of major bleeding in the EDX and WFR groups was 0.7% and 2.9%, respectively. The total incidence of major/minor bleeding in the EDX and WFR groups was 7.8% and 8.7%, respectively. Of note, the incidence of bleeding complications in the uninterrupted WFR strategy group was markedly high in patients with an estimated glomerular filtration rate (eGFR) <30 (75%) or a HAS-BLED score ≥3 (60%). Patients with eGFR ≥30 and a HAS-BLED score ≤2 had a lower incidence of bleeding (<10%), regardless of the administered anticoagulation drug (EDX or WFR). CONCLUSIONS: This study confirmed the safety and efficacy of uninterrupted anticoagulation therapy using EDX or WFR in real-world patients undergoing CA for AF. Patients with severely impaired renal function and/or a higher bleeding risk during uninterrupted therapy with WFR were at a prominent risk of bleeding. Therefore, particular attention should be paid in the treatment of these patients.

Predominant microvessel proliferation in coronary stent restenotic tissue in patients with diabetes: insights from optical coherence tomography image analysis.

BACKGROUND: Coronary optical coherence tomography (OCT) enables virtual depiction of histological findings of in-stent restenotic tissue. The aim of this study was to investigate the microvessel proliferation within in-stent restenotic tissue and the influence of diabetes mellitus (DM). METHODS: We examined 54 in-stent restenotic coronary artery lesions (stenotic area>50%) from 50 consecutive patients including 28 with DM (56%) and 9 insulin-treated DM patients (18%); who underwent coronary time-domain OCT imaging with automatic pull back (1mm/s, 20 frames/s). Microvessels were defined as low-signal cavities with a diameter of 50-150 microns and a trajectory parallel to the lumen recognized on 3 consecutive cross-sectional OCT image frames. The microvessel index was calculated as the number of frames with microvessel/total number of frames × 100. Patients were stratified into 3 groups: 1) without microvessels, 2) with a low (< median value) microvessel index, 3) with a high microvessel index. RESULTS: Microvessels were detected in 566 frames (3.1%) from 26 lesions (48%) in 24 patients (48%). A greater incidence of DM and higher serum glucose levels were observed in the high microvessel index group (DM: 42% vs 58% vs 83%, p=0.049; serum glucose level: 118.2 ± 44.6 vs 122.6 ± 31.0 vs 172.8 ± 63.1mg/dL, p=0.03 between low and high microvessel index group, p=0.005 between no microvessel and high microvessel index group). CONCLUSIONS: Microvessel formation may be a unique pathophysiological factor of in-stent restenoses in patients with DM.

Impact of combined assessment of coronary artery calcium score, carotid artery plaque score, and brachial-ankle pulse wave velocity for early coronary revascularization in patients with suspected coronary artery disease.

Coronary artery calcium score (CACS) measured by multi-detector computed tomography, carotid plaque score (CPS) measured by carotid artery ultrasound, and brachial-ankle pulse wave velocity (baPWV) are noninvasive screening tools for coronary artery disease. The aim of this study was to determine whether the combination of CACS, CPS, and baPWV improves the prognostic value for future cardiac events. CACS, CPS, and baPWV were assessed in 77 patients (mean age, 65 years, 49 males) undergoing invasive coronary angiography. ECG-triggered MSCT was used to assess CACS. CPS was defined as the sum of all plaque heights in bilateral carotid arteries. The highest baPWV was used for analysis. Cardiac events were defined as cardiac death, nonfatal myocardial infarction, or coronary revascularization. Thirty-two cardiac events (41.6%) occurred during follow-up (23.6 ± 20.8 months), consisting of 28 PCIs and 4 CABGs. The best cutoff values of positive CACS, CPS, and baPWV for predicting cardiac events were ≥ 50, ≥ 5, and ≥1.6 m/second, respectively. For the combination of the 3 modalities, the positive test was defined as having at least 1 positive result by each method. The negative predictive value of all 3 modalities combined was better than that of CACS alone. The event-free rate was higher in patients with negative results for all 3 parameters compared with those that were positive (100% versus 44.8%, P < 0.0001). The prognostic value of using combined assessment of CACS, CPS, and baPWV is more effective for predicting cardiac events than CACS alone.

Single-molecule observation of photocatalytic reaction in TiO2 nanotube: importance of molecular transport through porous structures.

We have investigated the photocatalytic activity of individual porous TiO(2) nanotubes by the single-molecule counting of (*)OH using a specific fluorescent probe. The time- and space-resolved observation of emissive fluorescein generated by the photocatalytic reaction clearly reveals the importance of the transport behavior of reagents through the porous structures on the photocatalytic activity and the existence of the spatial heterogeneity of reactive sites even in an isolated TiO(2) nanotube. An experiment on a single nanotube provided information that was useful for elucidating the reaction mechanism of the heterogeneous (photo)catalyst and for designing advanced porous materials.

Single-molecule detection of airborne singlet oxygen.

Airborne singlet oxygen (1O2) molecules, which are generated during the TiO2 photocatalytic reactions and diffused from the surface into air, were detected at the opposite surface using terrylenediimide (TDI) molecules at the single-molecule level. The novel 1O2 nanosensor, which has a detectable number of about 1000 1O2 molecules in 70 x 70 square micrometers, can easily detect the single 1O2 molecule at a distance of over 1000 micrometers from the place of its creation in ambient air.

Single-molecule fluorescence imaging of the remote TiO2 photocatalytic oxidation.

The remote TiO2 photocatalytic oxidation reaction of single dyes has been investigated by the single-molecule fluorescent imaging technique. The present results suggest that the active oxygen species (Ox) is most probably the .OH radical, which is generated from the photodecomposition of H2O2 by UV light. The analyses of the number, intensity, and spectrum of individual fluorescence spots at the single-molecule level also indicate that unoxidized and oxidized dyes exist during the bleaching processes of single dyes.