Optical Frequency Domain Imaging of Very Late Stent Thrombosis Following Bare-Metal Stent Implantation for Acute Myocardial Infarction.

A 43-year-old man was admitted to our hospital with ST-segment elevation acute coronary syndrome. He had experienced myocardial infarction 19 months previously, and a bare-metal stent (BMS) had been implanted in the culprit distal right coronary artery at another hospital. Emergency coronary angiography showed thrombotic in-stent occlusion of the BMS. Intravascular ultrasound revealed an undersized stent compared with the size of the vessel and late stent malapposition (LSM) with abundant thrombi. The lesion was successfully recanalized via thrombectomy and plain old balloon angioplasty. Optical frequency domain imaging performed at follow-up coronary angiography confirmed the improvement of the LSM and incomplete neointimal stent coverage. This report illustrates the importance of imaging modalities in elucidating the mechanism of BMS-related very late stent thrombosis.

Efficient isolation of human metapneumovirus using MNT-1, a human malignant melanoma cell line with early and distinct cytopathic effects.

Isolation of human metapneumovirus (HMPV) from clinical specimens is currently inefficient because of the lack of a cell culture system in which a distinct cytopathic effect (CPE) occurs. The cell lines LLC-MK2, Vero and Vero E6 are used for isolation of HMPV; however, the CPE in these cell lines is subtle and usually requires a long observation period and sometimes blind passages. Thus, a cell line in which an early and distinct CPE occurs following HMPV inoculation is highly desired by clinical virology laboratories. In this study, it was demonstrated that, in the human malignant melanoma cell line MNT-1, obvious syncytium formation occurs shortly after inoculation with HMPV-positive clinical specimens. In addition, the growth and efficiency of isolation of HMPV were greater using MNT-1 than using any other conventional cell line. Addition of this cell line to our routine viral isolation system for clinical specimens markedly enhanced isolation frequency, allowing isolation-based surveillance. MNT-1 has the potential to facilitate clinical and epidemiological studies of HMPV.

Novel full-spectral flow cytometry with multiple spectrally-adjacent fluorescent proteins and fluorochromes and visualization of in vivo cellular movement.

Flow cytometric analysis with multicolor fluoroprobes is an essential method for detecting biological signatures of cells. Here, we present a new full-spectral flow cytometer (spectral-FCM). Unlike conventional flow cytometer, this spectral-FCM acquires the emitted fluorescence for all probes across the full-spectrum from each cell with 32 channels sequential PMT unit after dispersion with prism, and extracts the signals of each fluoroprobe based on the spectral shape of each fluoroprobe using unique algorithm in high speed, high sensitive, accurate, automatic and real-time. The spectral-FCM detects the continuous changes in emission spectra from green to red of the photoconvertible protein, KikGR with high-spectral resolution and separates spectrally-adjacent fluoroprobes, such as FITC (Emission peak (Em) 519 nm) and EGFP (Em 507 nm). Moreover, the spectral-FCM can measure and subtract autofluorescence of each cell providing increased signal-to-noise ratios and improved resolution of dim samples, which leads to a transformative technology for investigation of single cell state and function. These advances make it possible to perform 11-color fluorescence analysis to visualize movement of multilinage immune cells by using KikGR-expressing mice. Thus, the novel spectral flow cytometry improves the combinational use of spectrally-adjacent various FPs and multicolor fluorochromes in metabolically active cell for the investigation of not only the immune system but also other research and clinical fields of use.

Transcatheter aspiration of a thrombus and percutaneous transluminal coronary recanalization for ST-segment elevation myocardial infarction related to coronavirus disease 2019.

Although the novel coronavirus disease 2019 (COVID-19) causes severe viral pneumonia, it has also been reported, in some cases, to co-exist with ST-segment elevation myocardial infarction. Here, we describe the case of a patient with COVID-19 and coronary risk factors for hypertension, including smoking and obesity, who developed acute myocardial infarction due to primary coronary artery thrombosis and was treated with transcatheter thrombus aspiration and percutaneous transluminal coronary recanalization (PTCR) with intracoronary urokinase administration. A large volume of thrombus was collected and thrombolysis in myocardial infarction flow grade 3 was obtained after the procedures. PTCR with or without transcatheter thrombus aspiration may be a useful treatment option. .

2-aminophenoxazine-3-one prevents pulmonary metastasis of mouse B16 melanoma cells in mice.

2-Aminophenoxazine-3-one (Phx-3) induced cellular apoptosis in mouse melanoma B16 cells as detected by DNA laddering and upregulated Fas expression in the cells in vitro. Next, the anti-metastatic effects of Phx-3 were investigated in C56BL/6 mice. When B16 melanoma cells were injected into the tail veins of mice, significant metastasis of the cells was indicated in the lungs, 14 days after treatment. In contrast, when 0.5 mg/kg Phx-3 was administered to mice through the tail veins, once simultaneously with or every three days after the administration of B16 melanoma cells, the number of metastasized pulmonary cells was extremely reduced. Moderate reduction of the number of metastasized pulmonary cells was indicated in the mice with a single dose of Phx-3 on day 3 after injection of the cells. However, when Phx-3 was administered in a single dose, 6 or 9 days after the injection of the cells, the number of metastasized pulmonary cells remained the same. The present results indicate that the metastasis of mouse B16 melanoma cells to the lung was significantly inhibited in mice administered Phx-3, which activated the intrinsic and extrinsic apoptotic pathways. The present study suggests that Phx-3 might be a potential anti-metastatic agent as well as an anticancer agent.

Acute myocardial infarction in a 25-year-old woman with sitosterolemia.

We report the case of acute myocardial infarction in a 25-year-old woman with sitosterolemia. She was treated using statins, but her low-density lipoprotein cholesterol (LDL-C) levels did not decrease appreciably. Genetic analysis revealed mutations in the ABCG8 gene. Ezetimibe treatment was initiated, and her LDL-C levels decreased substantially. Sitosterolemia must be considered in the differential diagnosis of familial hypercholesterolemia in case of early onset cardiovascular disease patient with high LDL-C.

Reproducibility of Brachial Artery Ultrasound Measurements.

Recent findings suggest that systemic artery endothelial function is associated with the preclinical phase of vascular disease and related to traditional atherosclerosis risk factors. Brachial artery diameter changes in response to hyperemia have been proposed recently as a noninvasive tool to assess endothelial function. To evaluate the reproducibility of brachial artery diameter measurements using ultrasound, we studied 12 healthy subjects (eight men and four women, mean age 37 +/- 9 years). An ATL HDI 3000 machine with a 5- to 10-MHz broadband transducer was used to image the right brachial artery at rest, approximately 4 cm above the elbow. Gray scale ultrasound and color Doppler long-axis images were recorded. Brachial arterial outer diameter (i.e., from anterior adventitia to posterior adventitia) and inner diameter (i.e., from anterior lumen-intima interface to posterior lumen-intima interface) were measured in each subject by two observers. An offline analysis system was used to make measurements at end-diastole from four cardiac cycles. Interobserver and intraobserver measurement variabilities (technical error rates) for brachial artery inner diameter were excellent, ranging from 2.5% to 3.8%. However, interobserver technical error rates for outer diameter measurements were significantly greater than those for inner diameter measurements, ranging from 16.3% to 22.1% (P < 0.001), presumably related to the difficulty in accurately defining the adventitial lines. There were no significant differences in interobserver and intraobserver variability for measurements made using gray scale and color Doppler-aided techniques. We conclude that interobserver and intraobserver reproducibility for brachial artery inner diameter measurements made from ultrasound images is excellent.

Programmable wireless light-emitting diode stimulator for chronic stimulation of optogenetic molecules in freely moving mice.

We produced a miniaturized, multicode, multiband, and programmable light-emitting diode (LED) stimulator for wireless control of optogenetic experiments. The LED stimulator is capable of driving three independent LEDs upon reception of an infrared (IR) signal generated by a custom-made IR transmitter. Individual LED photopulse patterns are assigned to different codes of the IR signals (up to 256 codes). The photopulse patterns can be programmed in the on-board microcontroller by specifying the parameters of duration ([Formula: see text]), frequency ([Formula: see text]), and pulse width ([Formula: see text]). The IR signals were modulated at multiple carrier frequencies to establish multiband IR transmission. Using these devices, we could remotely control the moving direction of a Thy1-ChR2-YFP transgenic mouse by transcranially illuminating the corresponding hemisphere of the primary motor cortex. IR transmitter and LED stimulator will be particularly useful in experiments where free movement or patterned concurrent stimulation is desired, such as testing social communication of rodents.

Tracking and quantification of dendritic cell migration and antigen trafficking between the skin and lymph nodes.

Skin-derived dendritic cells (DCs) play a crucial role in the maintenance of immune homeostasis due to their role in antigen trafficking from the skin to the draining lymph nodes (dLNs). To quantify the spatiotemporal regulation of skin-derived DCs in vivo, we generated knock-in mice expressing the photoconvertible fluorescent protein KikGR. By exposing the skin or dLN of these mice to violet light, we were able to label and track the migration and turnover of endogenous skin-derived DCs. Langerhans cells and CD103(+)DCs, including Langerin(+)CD103(+)dermal DCs (DDCs), remained in the dLN for 4-4.5 days after migration from the skin, while CD103(-)DDCs persisted for only two days. Application of a skin irritant (chemical stress) induced a transient >10-fold increase in CD103(-)DDC migration from the skin to the dLN. Tape stripping (mechanical injury) induced a long-lasting four-fold increase in CD103(-)DDC migration to the dLN and accelerated the trafficking of exogenous protein antigens by these cells. Both stresses increased the turnover of CD103(-)DDCs within the dLN, causing these cells to die within one day of arrival. Therefore, CD103(-)DDCs act as sentinels against skin invasion that respond with increased cellular migration and antigen trafficking from the skin to the dLNs.

Contrasting quiescent G0 phase with mitotic cell cycling in the mouse immune system.

A transgenic mouse line expressing Fucci (fluorescent ubiquitination-based cell-cycle indicator) probes allows us to monitor the cell cycle in the hematopoietic system. Two populations with high and low intensities of Fucci signals for Cdt1(30/120) accumulation were identified by FACS analysis, and these correspond to quiescent G0 and cycling G1 cells, respectively. We observed the transition of immune cells between quiescent and proliferative phases in lymphoid organs during differentiation and immune responses.

Isolated adrenocorticotropin deficiency associated with delirium and takotsubo cardiomyopathy.

We report a 65-year-old woman with isolated adrenocorticotropic hormone (ACTH) deficiency. The patient was transported to the emergency outpatient department by ambulance complaining of malaise and nausea. Because her laboratory data revealed hyponatremia, we performed endocrinological examinations and diagnosed isolated ACTH deficiency. After admission, she went into a delirious state and suffered from takotsubo cardiomyopathy due to adrenal insufficiency. Replacement therapy with hydrocortisone sufficiently improved her delirium and cardiomyopathy. We conclude that her unstable mental state and myocardial dysfunction were closely related to adrenal insufficiency and suggest that adrenal crisis may cause delirium and Takotsubo cardiomyopathy.

IFN-γ production downstream of NKT cell activation in mice infected with influenza virus enhances the cytolytic activities of both NK cells and viral antigen-specific CD8+ T cells.

Natural killer T (NKT) cell activation is responsible for eliminating pathogens. However, the biological functions of NKT cells against influenza virus are not fully understood. We therefore investigated the effects of NKT cells in viral infection using CD1d knockout (KO) mice. When CD1d KO or wild-type (WT) mice were infected with a sub-lethal dosage of the influenza virus, the survival rate of CD1d KO mice was significantly lower than for WT mice in association with delayed viral clearance in the lungs. Consistently, IFN-γ production in bronchoalveolar lavage fluid of CD1d KO mice was largely reduced compared to WT mice during infection. Moreover, the cytotoxic activities of NK cells and viral antigen-specific CD8(+) T cells were impaired in CD1d KO mice. It was concluded that activated NKT cell-induced IFN-γ release enhances both NK-cell activity and antigen-specific CD8(+) T cells to eliminate the influenza virus, thus leading to an enhanced survival.

Oxygen therapy prevents ventricular arrhythmias in patients with congestive heart failure and sleep apnea.

BACKGROUND: Although ventricular arrhythmia is critical for the prognosis of patients with severe congestive heart failure (CHF), it is difficult to control the arrhythmia using conservative therapies. However, many CHF patients also have sleep apnea syndrome (SAS) and oxygen supply improves their prognosis. The beneficial effects of oxygen treatment for ventricular arrhythmia have not yet been clarified, so the present study was designed to evaluate the effects of oxygen treatment for premature ventricular contraction (PVC). METHODS AND RESULTS: Patients with CHF and SAS were divided into 3 groups: (1) the "PVC declined" group that included patients who had frequent PVCs and oxygen treatment that suppressed the number of PVC; (2) the "PVC not affected" group that included patients who had frequent PVCs and oxygen treatment did not affect the number of PVC; and (3) the "few PVC" group that included patients who had no or few PVCs. The group 1 patients showed higher apnea-hypopnea index, standard deviation of all R-R intervals, left ventricular ejection fraction, and brain natriuretic peptide levels than the patients in group 2. Oxygen treatment in group 3 did not affect the PVC frequency. CONCLUSIONS: Oxygen treatment may be useful for preventing ventricular arrhythmia in selected patients with CHF and SAS.