A let-7 microRNA-RALB axis links the immune properties of iPSC-derived megakaryocytes with platelet producibility.

We recently achieved the first-in-human transfusion of induced pluripotent stem cell-derived platelets (iPSC-PLTs) as an alternative to standard transfusions, which are dependent on donors and therefore variable in supply. However, heterogeneity characterized by thrombopoiesis-biased or immune-biased megakaryocytes (MKs) continues to pose a bottleneck against the standardization of iPSC-PLT manufacturing. To address this problem, here we employ microRNA (miRNA) switch biotechnology to distinguish subpopulations of imMKCLs, the MK cell lines producing iPSC-PLTs. Upon miRNA switch-based screening, we find imMKCLs with lower let-7 activity exhibit an immune-skewed transcriptional signature. Notably, the low activity of let-7a-5p results in the upregulation of RAS like proto-oncogene B (RALB) expression, which is crucial for the lineage determination of immune-biased imMKCL subpopulations and leads to the activation of interferon-dependent signaling. The dysregulation of immune properties/subpopulations, along with the secretion of inflammatory cytokines, contributes to a decline in the quality of the whole imMKCL population.

[Left Ventricular Outflow Tract Obstruction After Transcatheter Aortic Valve Replacement due to Pannus Formation].

We experienced a case of surgical aortic valve re-replacement due to structural valve deterioration caused by pannus formation 4 years after transcatheter aortic valve replacement( TAVR). The patient underwent surgery because the mean transvalvular pressure gradient increased to 48 mmHg on echocardiography. Contrast-enhanced computed tomography (CT) was useful for predicting the site of adhesion to surrounding tissue preoperatively and exploring the presence of the pannus. Intraoperative findings showed the TAVR valve was covered with neointima except around the origins of the left and right coronary arteries and was firmly adhered to the surrounding tissues. As residual pannus was present in the subvalvular tissues, it was carefully removed. The explanted TAVR valve functioned well with good opening and closure. The postoperative course was uneventful. Pannus formation can result from mechanical stress. TAVR valves put significantly greater stress on the left ventricular outflow tract than surgical valves and may be more likely to cause pannus formation.

Remimazolam in perioperative management of Eisenmenger syndrome: a case report.

BACKGROUND: Eisenmenger syndrome (ES) is characterized by severe and irreversible pulmonary hypertension stemming from an uncorrected intracardiac shunt of significant size. The imbalance between systemic and pulmonary artery pressures predisposes patients with ES to the risk of cardiac arrest. Remimazolam has caused less circulatory depression, which may be advantageous for ES. However, no studies reported the use of remimazolam in perioperative ES management. CASE PRESENTATION: A 45-year-old female patient with ES derived from a ventricular septal defect was scheduled to undergo bilateral adnexectomy for an ovarian tumor. Her oxygen saturation was 80% with 3 L/min oxygen at rest, and her pulmonary and systemic flow ratio was 0.53. She underwent general anesthesia with remimazolam, and intraoperative hemodynamics was stable without hypotension or reduced oxygen saturation. CONCLUSIONS: Our successful management of ovarian tumor surgery in a patient with ES using remimazolam reveals its potential effectiveness in perioperative care.

[Surgical Explantation of a Bioprosthesis Nine Months After Transcatheter Aortic Valve Implantation].

CASE: 82-year-old female. In October 2020, the patient underwent transcatheter aortic valve implantation( TAVI)[Evolut PRO R 23 mm] via left subclavian artery approach for severe aortic valve stenosis. The patient was discharged home without any issues. However, nine months after surgery, the patient was hospitalized at another hospital for a right upper arm fracture and developed a fever. The patient was transferred to our hospital and was diagnosed with Enterococcal bacteremia while receiving antibiotic treatment. Echocardiography revealed leaflet thickening and cord-like structure on the artificial valve, and a diagnosis of prosthetic valve endocarditis (PVE) was made. The patient then underwent surgical removal of the valve. The patient had a good postoperative course and completed 4 weeks of antibiotic treatment before being transferred back to the referring hospital 31 days after surgery. TAVI made unprecendented revolution in the treatment of aortic valve stenosis. TAVI is often used for elderly patients with comorbidities because of high perioperative risk is for surgical aortic valve replacement. Although TAVI is widely promoted for its benefits, it is not without limitations. In cases with more than one year of follow-up, there are many complications and the risk of surgery is high. There have been few reports of cases from Japan requiring surgical removal of TAVI valve. In this case, PVE was diagnosed nine months after TAVI and the patient had a good outcome.

Successful lung-protective ventilatory management during the VV-ECMO in a severe COVID-19 pneumonia patient with extensive pneumomediastinum and subcutaneous emphysema: a case report.

BACKGROUND: Ventilatory management of respiratory failure with pneumomediastinum/subcutaneous emphysema is not established. Herein, we report a case of severe COVID-19 pneumonia with extensive pneumomediastinum/subcutaneous emphysema, rescued by thorough lung-protective ventilatory management after applying the VV-ECMO. CASE PRESENTATION: A 68-year-old male with no medical history was admitted to a local hospital and diagnosed with COVID-19 pneumonia. His pulmonary parameters worsened during invasive ventilation due to the development of pneumomediastinum/subcutaneous emphysema, and then he was transferred to our hospital. On arrival, we immediately decided to apply VV-ECMO and switch to ultraprotective ventilation. After maintaining the initial ventilation with a neuromuscular blocking agent for 2 days, we gradually increased PEEP while limiting PIP to 25 cmH2O. The patient was weaned off VV-ECMO on day 10; he was transferred to the medical ward after extubation. CONCLUSIONS: Lung-protective ventilatory management should be performed thoroughly during VV-ECMO in severe COVID-19 pneumonia with pneumomediastinum/subcutaneous emphysema.

A ß1-tubulin-based megakaryocyte maturation reporter system identifies novel drugs that promote platelet production.

During maturation, megakaryocytes (MKs) express ß1-tubulin (TUBB1) and rearrange their microtubule components to enlarge, form proplatelets, and eventually release platelets. The development of a platform to identify in vitro conditions that would efficiently promote MK development could potentially enable large-scale platelet production. Here, we show that an immortalized MK cell line (imMKCL) genetically modified to express the ß1-tubulin-Venus reporter provides a practical system to efficiently monitor the in vitro production of platelet-like particles (PLPs). The Venus transgene was inserted downstream of the TUBB1 locus in imMKCLs using CRISPR/Cas9, and the expression was visualized by Venus fluorescence intensity. This imMKCL reporter line was then used for high-throughput drug screening. We identified several compounds that significantly improved the efficiency of PLP production in vitro under feeder-free conditions and showed a significant tendency to recover platelets in vivo in a mouse thrombocytopenia model induced by anti-GPIbα antibody administration. Interestingly, most of these compounds, including a WNT signaling pathway inhibitor, Wnt-C59, antagonized the aryl hydrocarbon receptor (AhR) to increase PLP production, confirming the crucial role of AhR inhibition in MK maturation. Consistently, small interfering RNA treatment against AhR increased the Venus intensity and PLP production. TCS 359, an FLT3 inhibitor, significantly increased PLP production independently of FLT3 or AhR. This study highlights the usefulness of the ß1-tubulin reporter MK line as a useful tool to study the mechanisms underlying thrombopoiesis and to identify novel inducers of ex vivo platelet production.

Turbulence Activates Platelet Biogenesis to Enable Clinical Scale Ex Vivo Production.

The ex vivo generation of platelets from human-induced pluripotent cells (hiPSCs) is expected to compensate donor-dependent transfusion systems. However, manufacturing the clinically required number of platelets remains unachieved due to the low platelet release from hiPSC-derived megakaryocytes (hiPSC-MKs). Here, we report turbulence as a physical regulator in thrombopoiesis in vivo and its application to turbulence-controllable bioreactors. The identification of turbulent energy as a determinant parameter allowed scale-up to 8 L for the generation of 100 billion-order platelets from hiPSC-MKs, which satisfies clinical requirements. Turbulent flow promoted the release from megakaryocytes of IGFBP2, MIF, and Nardilysin to facilitate platelet shedding. hiPSC-platelets showed properties of bona fide human platelets, including circulation and hemostasis capacities upon transfusion in two animal models. This study provides a concept in which a coordinated physico-chemical mechanism promotes platelet biogenesis and an innovative strategy for ex vivo platelet manufacturing.

High electromechanical strain and enhanced temperature characteristics in lead-free (Na,Bi)TiO3-BaTiO3 thin films on Si substrates.

Here, we demonstrate the high electromechanical strain and enhanced temperature characteristics in the c-axis-oriented lead-free (Na,Bi)TiO3-BaTiO3 (NBT-BT) polycrystalline thin film prepared on Si substrates by rf magnetron sputtering. The effective transverse piezoelectric coefficient, e31*, estimated from the electromechanical strain measured under high electric field, reaches a high level of -12.5 C/m2, and is comparable to those of conventional Pb(Zr,Ti)O3 films. In-situ X-ray diffraction measurement and electron diffraction analysis revealed the electromechanical strain of the NBT-BT film to originate predominantly in elongation of the tetragonal (P4bm) crystal lattice in the c-axis direction. In addition to the large e31*, the NBT-BT film exhibits enhanced permittivity maximum temperature, Tm, of ~400 °C and no depolarization below Tm, as compared to bulk NBT-BT having Tm ≈ 300 °C and a depolarization temperature of ~100 °C. We conclude that the enhancement of temperature characteristics is associated with the distorted P4bm crystal lattice formed by deposition-induced stress and defects. We believe that the present study paves the way for practical applications of lead-free piezoelectric thin films in electromechanical devices.

Printable Organic Light-Emitting Devices and Application for Optical Signal Transmission.

Organic light emitting devices, in particular, properties of polymer light-emitting transistors with printed electrodes and bilayer printed devices with in-plane emission have been investigated and discussed. The semitransparent device based on poly(9,9-dioctylfluorene-co-benzothiadiazole) (F8BT) with Ag-nanowire source/drain and transparent carbon nanotube gate electrodes exhibits ambipolar and light-emitting characteristics. For the devices with oriented poly(9,9-dioctylfluorene) (F8) films, enhanced electron and hole field-effect mobilities have been achieved by aligning the polymer chains parallel to the transport direction. The bilayer device using F8BT lower layer and oriented F8 upper layer with the channel direction parallel to the polymer orientation exhibits improved EL intensity and higher external quantum efficiency than that with the channel direction perpendicular to the polymer chains orientation. The optical pulses of more than 100 Hz frequency are generated by directly modulating a bilayer device with an in-plane emission pattern.

Cell wall structure suitable for surface display of proteins in Saccharomyces cerevisiae.

A display system for adding new protein functions to the cell surfaces of microorganisms has been developed, and applications of the system to various fields have been proposed. With the aim of constructing a cell surface environment suitable for protein display in Saccharomyces cerevisiae, the cell surface structures of cell wall mutants were investigated. Four cell wall mutant strains were selected by analyses using a GFP display system via a GPI anchor. ß-Glucosidase and endoglucanase II were displayed on the cell surface in the four mutants, and their activities were evaluated. mnn2 deletion strain exhibited the highest activity for both the enzymes. In particular, endoglucanase II activity using carboxymethylcellulose as a substrate in the mutant strain was 1.9-fold higher than that of the wild-type strain. In addition, the activity of endoglucanase II released from the mnn2 deletion strain by Zymolyase 20T treatment was higher than that from the wild-type strain. The results of green fluorescent protein (GFP) and endoglucanase displays suggest that the amounts of enzyme displayed on the cell surface were increased by the mnn2 deletion. The enzyme activity of the mnn2 deletion strain was compared with that of the wild-type strain. The relative value (mnn2 deletion mutant/wild-type strain) of endoglucanase II activity using carboxymethylcellulose as a substrate was higher than that of ß-glucosidase activity using p-nitrophenyl-ß-glucopyranoside as a substrate, suggesting that the cell surface environment of the mnn2 deletion strain facilitates the binding of high-molecular-weight substrates to the active sites of the displayed enzymes.

Effect of steroids on increased permeability of blood vessels of the stria vascularis after auditory ossicle vibration by a drill in otologic surgery.

OBJECTIVES: The vibration caused by drills used for middle ear surgery is considered one of the causes of postoperative sensorineural deafness. Seki et al reported that when drill-induced damage was created in the auditory ossicles of guinea pigs, permeability across the capillary vessels in the stria vascularis increased significantly with the duration of drill-induced vibration. The present study was undertaken to examine changes in permeability across the stria vascularis capillaries following vibration in experimental animals pretreated with steroids, with the goal of developing a method of preventing a vibration-induced increase in permeability across these capillaries. METHODS: After an intravenous dose of hydrocortisone and horseradish peroxidase, the auditory ossicles of guinea pigs were vibrated with a drill for 60 seconds. RESULTS: Intravenous steroid administration before vibration reduced the leakage of horseradish peroxidase from the stria vascularis capillaries after vibration. CONCLUSIONS: The findings suggested that steroids suppress the increase in permeability across the stria vascularis capillaries that results from drill-induced vibration.

Caged gene-inducer spatially and temporally controls gene expression and plant development in transgenic Arabidopsis plant.

Two new types of caged gene-inducers, caged 17beta-estradiol and caged dexamethazone, were synthesized. Caged gene-inducers were applied to transgenic Arabidopsis plants carrying a steroid hormone-inducible transactivation system. Light uncaged caged gene-inducers and controlled spatial and temporal expression of transgene in the transgenic plant. Furthermore, caged gene-inducers enabled the control of root development by light.

Magnetic resonance imaging for diagnosis of congenital anosmia.

Magnetic Resonance Imaging (MRI) was performed on 9 patients who lacked a sense of smell since birth. Seven of them, including two patients with Kallmann syndrome, exhibited abnormality of the olfactory bulb, olfactory tract, olfactory sulcus, or rectus gyrus, with some variation among patients in type and degree of abnormality. The other two patients exhibited normal olfactory pathway morphology, and for them the possibility of acquired sensorineural anosmia could not be ruled out. MRI is useful for determining whether patients with congenital anosmia have olfactory pathway anomalies. Many patients with congenital anosmia and hypoplasty or aplasty of the olfactory pathway nevertheless had no gonadal or endocrinological disorders.

A case of fatal respiratory failure after surgery for advanced supraglottic laryngeal carcinoma.

Pulmonary lymphangitic carcinomatosis (PLC) is an uncommon type of pulmonary metastasis from laryngeal carcinoma. We encountered a case of fatal respiratory failure after surgery for supraglottic laryngeal carcinoma with multiple cervical lymph node metastases. On day 25 postoperatively, the patient suddenly developed fever and noted slowly progressive difficulty in breathing. Laboratory data, radiological findings, and the clinical course of this patient were strongly suggestive of PLC. The patient refused fiberoptic bronchoscopy and glucocorticoid therapy, leading to rapid deterioration due to increasing respiratory distress, and died of respiratory failure on postoperative day 32. PLC should be considered a possible presentation of pulmonary metastasis associated with head and neck carcinoma. Chest CT may be needed to detect subclinical PLC, especially in cases of advanced head and neck carcinomas.

[A case of advanced lateral wall oropharyngeal carcinoma showing complete response to chemotherapy with TS-1].

We report a recent case of squamous cell carcinoma originating in the right tonsil, with metastasis to the right superior cervical lymph nodes (T3N2bM0: stage IVA). Chemo-radiotherapy was administered as first-line therapy; and adjuvant chemotherapy with TS-1 was applied during the subsequent 4-week period for the treatment of the residual tumor in the neck. In this patient (female, 61 years old), the primary tonsillar tumor showed a complete response to the first-line therapy, however, the metastatic lesions in the cervical lymph nodes persisted, although the nodes also decreased markedly in size. Subsequent chemotherapy with TS-1 (80 mg/day) given for a 4-week period resulted in complete disappearance of the residual malignancy (CR), as determined by palpation and diagnostic imaging. No serious adverse events occurred during the therapy. Surgical treatment in patients with advanced head and neck cancer often results in a diminished quality of life. Although further accumulation of cases is necessary, TS-1, which allows oral chemotherapy on an outpatient basis, would seem to be a useful drug for adjuvant chemotherapy following radical irradiation in patients with advanced head and neck cancer.