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Could enzymatic activities and their cooperative functions act as cellular temperature-sensing systems? This review introduces recent opto-thermal technologies for microscopic analyses of various types of cellular temperature-sensing system. Optical microheating technologies have been developed for local and rapid temperature manipulations at the cellular level. Advanced luminescent thermometers visualize the dynamics of cellular local temperature in space and time during microheating. An optical heater and thermometer can be combined into one smart nanomaterial that demonstrates hybrid function. These technologies have revealed a variety of cellular responses to spatial and temporal changes in temperature. Spatial temperature gradients cause asymmetric deformations during mitosis and neurite outgrowth. Rapid changes in temperature causes imbalance of intracellular Ca2+ homeostasis and membrane potential. Among those responses, heat-induced muscle contractions are highlighted. It is also demonstrated that the short-term heating hyperactivates molecular motors to exceed their maximal activities at optimal temperatures. We discuss future prospects for opto-thermal manipulation of cellular functions and contributions to obtain a deeper understanding of the mechanisms of cellular temperature-sensing systems.

Drebrin E is a regulatory protein of intracellular force produced by actomyosin complexes, that is, myosin molecular motors interacting with actin filaments. The expression level of drebrin E in nerve cells decreases as the animal grows, suggesting its pivotal but unclarified role in neuronal development. Here, by applying the microscopic heat pulse method to actomyosin motility assay, the regulatory mechanism is examined from the room temperature up to 37 °C without a thermal denaturing of proteins. We show that the inhibition of actomyosin motility by drebrin E is eliminated immediately and reversibly during heating and depends on drebrin E concentration. The direct observation of quantum dot-labeled drebrin E implies its stable binding to actin filaments during the heat-induced sliding. Our results suggest that drebrin E allosterically modifies the actin filament structure to regulate cooperatively the actomyosin activity at the maintained in vivo body temperature., 胎児の神経を形作る仕組みは精密な温度センサー --母体の体温維持が神経の成熟に重要であることを示唆--. 京都大学プレスリリース. 2021-11-10.

Could enzymatic activities and their cooperative functions act as cellular temperature-sensing systems? This review introduces recent opto-thermal technologies for microscopic analyses of various types of cellular temperature-sensing system. Optical microheating technologies have been developed for local and rapid temperature manipulations at the cellular level. Advanced luminescent thermometers visualize the dynamics of cellular local temperature in space and time during microheating. An optical heater and thermometer can be combined into one smart nanomaterial that demonstrates hybrid function. These technologies have revealed a variety of cellular responses to spatial and temporal changes in temperature. Spatial temperature gradients cause asymmetric deformations during mitosis and neurite outgrowth. Rapid changes in temperature causes imbalance of intracellular Ca2+ homeostasis and membrane potential. Among those responses, heat-induced muscle contractions are highlighted. It is also demonstrated that the short-term heating hyperactivates molecular motors to exceed their maximal activities at optimal temperatures. We discuss future prospects for opto-thermal manipulation of cellular functions and contributions to obtain a deeper understanding of the mechanisms of cellular temperature-sensing systems.

According to current short-term evidence, ventricular septal defect (VSD) closure should be performed as early as possible after aortic regurgitation (AR) diagnosis in pediatric patients to prevent AR progression. However, long-term follow-up data are lacking. Therefore, our aim was to evaluate the long-term follow-up (≥ 10 years) of patients who underwent VSD closure as early as possible after AR diagnosis and to evaluate whether early VSD closure prevents progression of AR. This was a retrospective cohort study of 42 patients with VSD and AR who had undergone VSD closure at a median age of 2.9 years, with a median waiting period from AR diagnosis to VSD closure of 3.4 months and follow-up of a median 13.1 years (interquartile range 10.0-15.8 years). The preoperative degree of AR was trivial in 25 patients, mild in 15, and moderate in 2. Of the 33 patients followed up for ≥ 10 years, none required aortic valve replacement and there was no incidence of mortality. The degree of AR improved or did not change, except in 1 patient. The size of the VSD (odds ratio [OR] 0.85; 95% confidence interval [CI] 0.62-1.18; p = 0.33), time from diagnosis to surgery (OR 1.00; 95% CI 1.000-1.001; p = 0.657), and age at the time of surgery (OR 1.00; 95% CI 0.998-1.004; p = 0.452) were not predictive of persistent postoperative AR. Therefore, VSD closure performed as early as possible after AR diagnosis could successfully prevent AR progression in patients with less than moderate preoperative AR, eliminating the need for aortic valve replacement and valvuloplasty.

Background Mutations in GNAO1 typically result in neurodevelopmental disorders, including involuntary movements. They may be improved using calcium-channel modulators. Case The patient visited our hospital at age 2 years because of moderate global developmental delay. Her intermittent, generalized involuntary movements started at age 8 years. A de novo GNAO1 mutation, NM_020988.2:c.626G > A, (p.Arg209Cys), was identified by whole exome sequencing. At age 9 years, she experienced severe, intermittent involuntary movements, which led to rhabdomyolysis. She needed intensive care with administration of midazolam, dantrolene sodium hydrate, and plasma exchange. We started treating her with gabapentin (GBP), after which she recovered completely. At age 11 years, she developed continuous, generalized involuntary movements. This prompted us to increase the GBP dose, which again resolved the involuntary movements completely. Conclusion In the case of movement disorders associated with GNAO1 mutations, GBP treatment may be attempted before more invasive procedures are performed.

The patient was a 28-year-old woman who was misdiagnosed with tetralogy of Fallot and straddling mitral valve after birth. She underwent a left modified Blalock-Taussig shunt at the age of 1 year. At age 28, she presented with fatigue and progressive cyanosis. Finally, she was diagnosed with tetralogy of Fallot and complete atrioventricular septal defect. To measure the exact biventricular volumes, we performed cardiac magnetic resonance imaging in addition to cardiac angiography and ensured adequate volume capacity. We eventually decided to perform biventricular repair. Her postoperative course was uneventful, and she returned to full-time work.

Introduction A late aortic aneurysm at the site of previous open surgery for coarctation of the aorta (CoA) is a life threatening complication. Several strategies have been reported, however, these strategies have issues. This is the report of a case involving successful ascending to descending aortic bypass concomitant with endovascular embolisation for a pseudoaneurysm. Report The patient was a 23 year old man with a history of patch aortoplasty for coarctation of the aorta (CoA) via a left thoracotomy and patch closure of a ventricular septal defect. Enhanced computed tomography (CT) angiography performed in 2014 revealed a pseudoaneurysm at the site of the previous patch aortoplasty. CT also revealed a hypoplastic and severely kinked aortic arch that made it difficult to perform conventional thoracic endovascular aortic repair for this aneurysm. Therefore in order to prevent rupture, a combination of ascending to descending aortic bypass and endovascular embolisation using Amplatzer vascular plugs and coils was employed. Transection of the aortic arch and transposition of the left subclavian artery were performed to prevent antegrade flow into the aneurysm. Discussion The aneurysm was successfully excluded without complications. CT performed three years after the operation showed that the pseudoaneurysm had shrunk completely. Extra-anatomic bypass concomitant with endovascular embolisation for a late coarctation patched site aneurysm might be an alternative strategy that should be carefully considered in specific cases., Highlights • A case of late coarctation patched site pseudoaneurysm is presented. • Ascending to descending aortic bypass was performed successfully. • The aneurysm was treated successfully by endovascular embolisation. • Follow up computed tomography showed that the pseudoaneurysm had shrunk completely. • This approach might be an alternative strategy that should be carefully considered.

Type 1 ryanodine receptor (RYR1) is a Ca2+ release channel in the sarcoplasmic reticulum (SR) of the skeletal muscle and plays a critical role in excitation–contraction coupling. Mutations in RYR1 cause severe muscle diseases, such as malignant hyperthermia, a disorder of Ca2+-induced Ca2+ release (CICR) through RYR1 from the SR. We recently reported that volatile anesthetics induce malignant hyperthermia (MH)-like episodes through enhanced CICR in heterozygous R2509C-RYR1 mice. However, the characterization of Ca2+ dynamics has yet to be investigated in skeletal muscle cells from homozygous mice because these animals die in utero. In the present study, we generated primary cultured skeletal myocytes from R2509C-RYR1 mice. No differences in cellular morphology were detected between wild type (WT) and mutant myocytes. Spontaneous Ca2+ transients and cellular contractions occurred in WT and heterozygous myocytes, but not in homozygous myocytes. Electron microscopic observation revealed that the sarcomere length was shortened to ∼1.7 µm in homozygous myocytes, as compared to ∼2.2 and ∼2.3 µm in WT and heterozygous myocytes, respectively. Consistently, the resting intracellular Ca2+ concentration was higher in homozygous myocytes than in WT or heterozygous myocytes, which may be coupled with a reduced Ca2+ concentration in the SR. Finally, using infrared laser-based microheating, we found that heterozygous myocytes showed larger heat-induced Ca2+ transients than WT myocytes. Our findings suggest that the R2509C mutation in RYR1 causes dysfunctional Ca2+ dynamics in a mutant-gene dose-dependent manner in the skeletal muscles, in turn provoking MH-like episodes and embryonic lethality in heterozygous and homozygous mice, respectively.

私たちの体の中には、臓器が正常に機能するに不可欠なヒダや突起が存在する。しかし、細胞を従来から用いられている硬いプラスチック皿で培養しても平面状に広がるだけで、体の中の立体構造がいかにして形作られるのか、未だ詳細が分かっていない。そこで細胞の力に注目し、細胞が接着面を引っ張る小さな力でも変形するフレキシブル薄膜をイオンビーム加工技術で開発した。薄膜の厚みやパターンの制御により、培養細胞に薄膜を変形させて、ヒダや突起構造を形作らせることに成功した。, QST高崎サイエンスフェスタ2021

ディッシュ上で培養した細胞は平面状に広がるだけで、生体内の細胞がどのようにして立体構造を形作るのか、未だによく分かっていない。我々は、細胞の力で変形する培養用薄膜があれば、細胞が立体構造を作るのではないかと考えた。イオンビームがポリ乳酸中に誘起する分解と架橋の割合を空間的に制御することで、細胞の小さな力で剥離・変形可能な薄膜を開発し、細胞にヒダや突起を作らせて３次元細胞シートを得ることに成功した。, 日本原子力学会 関東・甲越支部 第20回 若手研究者・技術者発表討論会

人工血管移植後の漿液腫は、入院期間の延長や再手術のみならず感染合併により最悪肢切断に至る重篤な合併症である。移植後の周囲器質化を促進させ漿液腫を予防するために、ガンマ線架橋ゼラチンハイドロゲル(RXゲル)を人工血管外側に被覆した。RXゲルは細胞外マトリックスに極めて近い組成・物性を有し、ゲル硬度や含水度を制御することができる。RXゲル被覆人工血管を動物に移植して評価したところ、従来の人工血管に比べ器質化が促進されていることが分かったので報告する。, 第62回日本脈管学会総会

Sarcomeric contraction in cardiomyocytes serves as the basis for the heart’s pump functions. It has generally been considered that in cardiac muscle as well as in skeletal muscle, sarcomeres equally contribute to myofibrillar dynamics in myocytes at varying loads by producing similar levels of active and passive force. In the present study, we expressed α-actinin–AcGFP in Z-disks to analyze dynamic behaviors of sequentially connected individual sarcomeres along a myofibril in a left ventricular (LV) myocyte of the in vivo beating mouse heart. To quantify the magnitude of the contribution of individual sarcomeres to myofibrillar dynamics, we introduced the novel parameter “contribution index” (CI) to measure the synchrony in movements between a sarcomere and a myofibril (from −1 [complete asynchrony] to 1 [complete synchrony]). First, CI varied markedly between sarcomeres, with an average value of ∼0.3 during normal systole. Second, when the movements between adjacent sarcomeres were asynchronous (CI < 0), a sarcomere and the ones next to the adjacent sarcomeres and farther away moved in synchrony (CI > 0) along a myofibril. Third, when difference in LV pressure in diastole and systole (ΔLVP) was lowered to

Complex geometries such as folds and protrusions in various organs are formed by the wrinkling/buckling of flat epithelial sheets. This morphogenesis is considered to be triggered by mechanical instabilities between the epithelial sheet and the mesenchyme foundation, similar to that observed in non-living film-on-substrate structures. Here, we report a film-on-substrate cell culture system (FoSS), in which the cells can induce wrinkling and buckling-delamination of the surface layer from the foundation. Ion beam irradiation on polylactic acid (PLA) produced three layers: a crosslinked/carbonized surface, a decomposed middle layer that undergoes solation under the cell culture condition, and a foundation. This system was realized by controlling the three-dimensional (3D) distributions of scission/crosslinking probabilities in PLA. Madin-Darby canine kidney epithelial cells adhered well to the surface layer of FoSS. Cellular forces caused the layer to become wrinkled and subsequently delaminated as a thin film (~165 nm) from the foundation. The cell population then crumpled the peeled films, and within 48 h formed 3D cell sheets with folds and protrusions depending on patterns pre-set on the FoSS. FoSS can be utilized to produce in vitro models of 3D morphogenesis and 3D cell sheets that are currently in demand for tissue engineering and regenerative medicine.

The predictive ability of neonatal illness severity scores for mortality or morbidity in extremely premature infants has not been extensively studied. We aimed to evaluate the ability of neonatal illness severity scores [Clinical Risk Index for Babies II (CRIB II), Score for Neonatal Acute Physiology II (SNAP-II), and SNAP-Perinatal Extension II (SNAPPE-II)] in predicting mortality and short-term morbidity of extremely premature infants.This retrospective study involved 171 infants with gestational age (GA) between 22 and 27 weeks who were admitted to the NICU during 2010-2017. Predictive ability of neonatal illness severity scores for mortality and short-term morbidity (bronchopulmonary dysplasia, retinopathy of prematurity, intraventricular hemorrhage, necrotizing enterocolitis, and gastrointestinal perforation) was assessed by comparing their area under the receiver operating characteristic curve.The overall mortality rate was 11.1%. Mortality at 23 weeks' gestation was higher than that at 24-27 weeks' gestation (The neonatal illness severity scores were excellent predictors of early death in extremely premature infants and might be useful for selecting extremely preterm infants who need intervention.

Background Overnight catheter drainage (OCD) is introduced to avoid overdistention of the bladder at night-time when clean intermittent catheterization proves ineffective for daytime management of neurogenic bladder. We adopted OCD using disposable silicone no-balloon (DSnB) catheters, with the distal end outside the body opening into diapers. OCD using DSnB catheter, however, had risks of retrograde bacterial contamination. Therefore, in this study, the validity of equipping DSnB catheters with check valves to prevent retrograde bacterial contamination was examined. Methods For the in vitro study, excised saline-filled swine bladders were drained using DSnB catheters with or without check valves, and the time required for intravesical pressure to reach 5 cmH2 O was measured. For the in vivo study, in cross-over experiments comparing DSnB catheters with and without check valves, OCD using DSnB catheters for 10 h was performed in rabbits under analgesia. Bacterial growth from urine samples before and after OCD and residual urine volume were examined. Results For the in vitro experiment, the median drainage time was 368.2 s (range, 88-1,085 s) and 344.7 s (range, 28-840 s) with and without check valves, respectively (n = 6), which was not significantly different. For the in vivo experiment, in cross-over experiments (n = 8) new bacterial growth rates after OCD did not differ, and median residual urine volume was 17.1 mL (range, 0-75 mL) and 1.2 mL (range, 0-5 mL) with and without check valves, respectively (P = 0.055). Conclusions Installing a check valve in the DSnB catheter did not decrease new bacterial growth, while tending to increase residual urine volume. DSnB catheters without check valves appear to be better for continuous drainage of urine from bladder.

Muscle fiber contraction involves chemical reactions and structural changes that are sensitive to temperature. Ishii et al. show that rapid heating causes cardiac thin filament sliding, and the data could indicate that thin filaments are partially activated during diastole at mammalian body temperature., During the excitation–contraction coupling of the heart, sarcomeres are activated via thin filament structural changes (i.e., from the “off” state to the “on” state) in response to a release of Ca2+ from the sarcoplasmic reticulum. This process involves chemical reactions that are highly dependent on ambient temperature; for example, catalytic activity of the actomyosin ATPase rises with increasing temperature. Here, we investigate the effects of rapid heating by focused infrared (IR) laser irradiation on the sliding of thin filaments reconstituted with human α-tropomyosin and bovine ventricular troponin in an in vitro motility assay. We perform high-precision analyses measuring temperature by the fluorescence intensity of rhodamine-phalloidin–labeled F-actin coupled with a fluorescent thermosensor sheet containing the temperature-sensitive dye Europium (III) thenoyltrifluoroacetonate trihydrate. This approach enables a shift in temperature from 25°C to ∼46°C within 0.2 s. We find that in the absence of Ca2+ and presence of ATP, IR laser irradiation elicits sliding movements of reconstituted thin filaments with a sliding velocity that increases as a function of temperature. The heating-induced acceleration of thin filament sliding likewise occurs in the presence of Ca2+ and ATP; however, the temperature dependence is more than twofold less pronounced. These findings could indicate that in the mammalian heart, the on–off equilibrium of the cardiac thin filament state is partially shifted toward the on state in diastole at physiological body temperature, enabling rapid and efficient myocardial dynamics in systole.

We describe a seven-month-old boy with tetralogy of Fallot and an absent left pulmonary artery. Due to the diminutive size of the left pulmonary artery, we performed a native tissue left pulmonary artery reconstruction and intrapulmonary artery septation procedure with a left modified Blalock-Taussig shunt. After confirming left pulmonary artery growth, the patient underwent tetralogy of Fallot repair, removal of septation patch, and division of the Blalock-Taussig shunt. Nine months post-surgery, we confirmed his balanced lung perfusion (R/L ratio 6:4). The intrapulmonary artery septation procedure would be suitable for both the resuscitation and reconstruction of the hypoplastic absent pulmonary artery.

Objective: To compare left ventricular (LV) and right ventricular (RV) volume, function, and image quality of a respiratory-triggered two-dimensional (2D)-cine k-adaptive-t-autocalibrating reconstruction for Cartesian sampling (2D kat-ARC) with those of the standard reference, namely, breath-hold 2D balanced steady-state free precession (2D SSFP), in patients with repaired tetralogy of Fallot (TOF). Methods: 30 patients (14 males, mean age 32.2 ± 13.9 years) underwent cardiac magnetic resonance, and 2D kat-ARC and 2D SSFP images were acquired on short-axis view. Biventricular end-diastolic volume (EDV) and end-systolic volume (ESV), stroke volume (SV), ejection fraction (EF), and LV mass (LVM) were analysed. Results: The 2D kat-ARC had significantly shorter scan time (35.2 ± 9.1 s vs 80.4 ± 16.7 s; p < 0.0001). Despite an analysis of image quality showed significant impairment using 2D kat-ARC compared to 2D SSFP cine (p < 0.0001), the two sequences demonstrated no significant difference in terms of biventricular EDV, LVESV, LVSV, LVEF, and LVM. However, the RVESV was overestimated for 2D kat-ARC compared with that for 2D SSFP (73.8 ± 43.2 ml vs 70.3 ± 44.5 ml, p = 0.0002) and the RVSV and RVEF were underestimated (RVSV = 46.2±20.5 ml vs 49.4 ± 20.4 ml, p = 0.0024; RVEF = 40.2±12.7% vs. 43.5±14.0%, p = 0.0002). Conclusion: Respiratory-triggered 2D kat-ARC cine is a reliable technique that could be used in the evaluation of LV volumes and function. Advances in knowledge: 2D cine kat-ARC is a reliable technique for the assessment LV volume and function in patients with repaired TOF.

量子ビーム橋かけ技術を駆使してタンパク質をゲル化し、さらにその硬さや形状を制御する新技術を実現した。このゲルはタンパク質本来の生体適合性や生分解性を保持しており、硬さを軟組織全域で自在に調整でき、なおかつ表面に微小な凹凸構造を付与することもできる。生体内における細胞外マトリックスの化学的・物理的特性を再現したゲルの上では、ディッシュ上では見ることができない３次元的な細胞の姿や機能発現を見ることができる。, 第20回日本再生医療学会総会

Hydrostatic pressure control has attracted much attention and presents a still challenging objective from mechanobiological viewpoints. Herein, we reveal the calcium entry processes in HeLa cells by means of hydrostatic pressure spectroscopy. The steady-state fluorescence spectral data comprehensively elucidated the factors controlling the outcomes of the hydrostatic pressure-stimulated calcium entry behavior. The present work leads to a new perspective on ion regulations in living cells and an attractive alternative to conventional mechanostimuli.

Turner syndrome (TS) is the most frequent sex abnormality in women. The physical features include short stature, webbing of the neck, and gonadal dysgenesis. Typically, patients with Turner syndrome exhibit no intellectual disability, and a few cases of TS have been associated with epilepsy. Herein, we present a case of TS with intractable epilepsy. The patient presented with global developmental delay at the age of two and karyotyping revealed mosaicism [45, X/46, X del (X) (q21.1)]. At the age of seven, she had generalized tonic epilepsy as well as several focal-onset seizures. She developed daily seizures, which were refractory to several antiepileptic drugs. Interictal electroencephalography (EEG) revealed multifocal spikes, and ictal EEG revealed shifting foci. She visited our hospital at the age of 13. Her peripheral white blood cells G-band and fluorescence in situ hybridization (FISH) method chromosome with cheek swab examinations revealed 45, X. Her peripheral white blood cell mosaic pattern may have disappeared over time or become indetectable. We treated her with clobazam, and then lamotrigine and valproic acid combination therapy, which resulted in a reduction in the frequency of seizures by approximately 50%. Epilepsy and intellectual disability in this case may be due to the mosaic deletion at Xq21.1. Further analysis of similar cases may provide valuable information for effective therapeutic strategies.

The elasticity, topography, and chemical composition of cell culture substrates influence cell behavior. However, the cellular responses toin vivoextracellular matrix (ECM), a hydrogel of proteins (mainly collagen) and polysaccharides, remain unknown as there is no substrate that preserves the key features of native ECM. This study introduces novel collagen hydrogels that can combine elasticity, topography, and composition and reproduce the correlation between collagen concentration (C) and elastic modulus (E) in native ECM. A simple reagent-free method based on radiation-cross-linking altered ECM-derived collagen I and hydrolyzed collagen (gelatin or collagen peptide) solutions into hydrogels with tunable elastic moduli covering a broad range of soft tissues (E= 1-236 kPa) originating from the final collagen density in the hydrogels (C= 0.3%-14%) and precise microtopographies (⩾1 μm). The amino acid composition ratio was almost unchanged by this method, and the obtained collagen hydrogels maintained enzyme-mediated degradability. These collagen hydrogels enabled investigation of the responses of cell lines (fibroblasts, epithelial cells, and myoblasts) and primary cells (rat cardiomyocytes) to soft topographic cues such as thosein vivounder the positive correlation betweenCandE. These cells adhered directly to the collagen hydrogels and chose to stay atop or spontaneously migrate into them depending onE, that is, the density of the collagen network,C. We revealed that the cell morphology and actin cytoskeleton organization conformed to the topographic cues, even when they are as soft asin vivoECM. The stiffer microgrooves on collagen hydrogels aligned cells more effectively, except HeLa cells that underwent drastic changes in cell morphology. These collagen hydrogels may not only reducein vivoandin vitrocell behavioral disparity but also facilitate artificial ECM design to control cell function and fate for applications in tissue engineering and regenerative medicine.

Background: Left ventricular noncompaction (LVNC) is a hereditary type of cardiomyopathy. Although it is associated with high morbidity and mortality, the related ion channel gene variants in children have not been fully investigated. This study aimed to elucidate the ion channel genetic landscape of LVNC and identify genotype-phenotype correlations in a large Japanese cohort. Methods: We enrolled 206 children with LVNC from 2002 to 2017 in Japan. LVNC was classified as follows: LVNC with congenital heart defects, arrhythmia, dilated phenotype, or normal function. In the enrolled patients, 182 genes associated with cardiomyopathy were screened using next-generation sequencing. Results: We identified 99 pathogenic variants in 40 genes in 87 patients. Of the pathogenic variants, 8.8% were in genes associated with channelopathies, 27% were in sarcomere genes, and 11.5% were in mitochondrial genes. Ion channel gene variants were mostly associated with the arrhythmia classification, whereas sarcomere and mitochondrial gene variants were associated with the dilated phenotype. Echocardiography revealed that the group with ion channel gene variants had almost normal LV ejection fraction and LV diastolic diameter Z scores. Fragmented QRS, old age, and an arrhythmia phenotype were the most significant risk factors for ventricular tachycardia ( P =0.165, 0.0428, and 0.0074, respectively). Moreover, the group with ion channel variants exhibited a greater risk of a higher prevalence of arrhythmias such as ventricular tachycardia, rather than congestive heart failure. Conclusions: This is the first study that focused on genotype-phenotype correlations in a large pediatric LVNC patient cohort with ion channel gene variants that were determined using next-generation sequencing. Ion channel gene variants were strongly correlated with arrhythmia phenotypes. Genetic testing and phenotype specification allow for appropriate medical management of specific LVNC targets.

Aims Critical congenital heart disease (CCHD) requires surgery or catheter intervention within the first year of life; delayed diagnoses result in worsened outcomes. In Japan, there are few reports of delayed CCHD diagnosis. We investigated the diagnoses and factors associated with the late detection of CCHD. Methods This retrospective cohort study evaluated 88 CCHD infants admitted to a level IV facility. We compared the late detection rates across facility levels of neonatal care and CCHD characteristics. Results Critical congenital heart disease was identified prenatally in 46 (52%) infants: early (≤3 days postnatally) in 29 (33%) and late (≥4 days postnatally) in 13 (15%). The oxygen saturation of 27 of 29 infants with early detection and 7 of 13 infants with late detection was measured using pulse oximetry within 3 days postnatally. Factors associated with the late detection of CCHD were lack of clinical recognition of symptoms in five infants, referral to higher-level facilities after discharge in four infants, and definitive diagnosis not confirmed using echocardiography in four infants. The most common factors associated with late detection were referral to higher-level facilities after discharge in level I facilities and definitive diagnosis not confirmed using echocardiography in level II facilities. Conclusion Critical congenital heart disease may require advanced knowledge and echocardiographic techniques for diagnosis. Pulse oximetry and telemedicine should also be incorporated in the diagnostic algorithm. Improvement in these factors might contribute to reducing the late detection of CCHD.

Polydimethylsiloxane (PDMS) has many desirable features for microfluidics applications, particularly in diagnostics and pharmaceuticals, but its hydrophobicity and the lack of a practical method for bonding PDMS layers limit its use. Moreover, the flexibility of PDMS causes unwanted deformation during use in some applications. Here, we report a simple method for solving these problems simultaneously using an electron beam (EB) or γ-rays, which are commonly used for sterilizing medical products. Simply by applying EB or γ-ray irradiation to stacked PDMS layers, we can not only bond the interfaces between the layers by forming Si–O–Si covalent bonds but also achieve long-lasting hydrophilization and sterilization of the internal microchannels and chambers, prevent nonspecific adsorption and absorption of hydrophobic small molecules, and enhance the mechanical strength of the material by converting bulk PDMS into a Si–Ox-rich (where x is 3 or 4) structure though crosslinking. Unlike the one-at-a-time plasma process, EBs and γ-rays can penetrate through many stacked layers of PDMS sealed in their final package, enabling batch modification and bonding. The method requires no chemical crosslinkers, adhesive agents, or fillers; hence, it does not undermine the advantages of PDMS such as ease of molding in soft lithography, biocompatibility, and optical transparency. Furthermore, bonding is achieved with high-throughput yield because it occurs after re-adjustable alignment. We demonstrate that this method is applicable in the mass production of 3D integrated PDMS microfluidic chips with some glass-like properties as well as for 3D structures with complex shapes that are difficult to fabricate with plastic or glass.

腹部手術後に起こる周囲臓器との癒着は患者のQOLを著しく低下させる炎症反応であり、手術時に体内に留置する癒着防止剤の開発が望まれている。本研究では、細胞非接着材料のカルボキシメチルセルロース(CMC)と生体適合性・生分解性に優れたゼラチンからなるハイブリッドゲルをガンマ線架橋技術で創出した。架橋剤等を用いずに作製したこのゲルは、母材の生体適合性・生分解性を保っている。成分比を調整した結果、CMC60%・ゼラチン40%の割合で作製したゲルは、線維芽細胞の接着・浸潤を防止しつつ、細胞生存率を有意に高めることが分かった。また、このゲルの生分解速度を調整すると共に、硬さを腹部と同程度となる数10kPaの圧縮弾性率に調整することもできた。移植物と臓器との機械的なマッチングは異物反応を抑制することが分かっており、開発したゲルは、移植後の異物反応を抑制し創傷治癒後に分解される、機能性癒着防止剤として応用が期待される。

Vitamin K deficiency in pregnant women causes intracranial hemorrhage (ICH) in fetuses. Fetal ICH frequently causes life-threatening and persistent neurological damage. However, indicators for preventing fetal ICH are not established. Two pregnant women developed long-term eating disorders caused by psychosis. They were administered intravenous fluid and vitamin supplementation, excluding vitamin K. The intracranial low-hypoechoic area on fetal ultrasound was suggestive of fetal ICH due to vitamin K deficiency. Their neonates showed severe developmental delay. Laboratory analysis revealed a normal prothrombin time, but elevated protein induced by vitamin K absence II. Pregnant women who have eating disorders more than 3 weeks could develop fetal ICH due to maternal subclinical vitamin K deficiency. Illness duration and protein induced by vitamin K absence II of pregnant woman may be indicators for vitamin K administration to prevent fetal intracranial hemorrhage.

The present study was conducted to systematically investigate the optimal viral titer as well as the volume of the adenovirus vector (ADV) that expresses α-actinin-AcGFP in the Z-disks of myocytes in the left ventricle (LV) of mice. An injection of 10 μL ADV at viral titers of 2 to 4 × 1011 viral particles per mL (VP/mL) into the LV epicardial surface consistently expressed α-actinin-AcGFP in myocytes in vivo, with the fraction of AcGFP-expressing myocytes at ~10%. Our analysis revealed that SL was ~1.90-2.15 μm upon heart arrest via deep anesthesia. Likewise, we developed a novel fluorescence labeling method of the T-tubular system by treating the LV surface with CellMask Orange (CellMask). We found that the T-tubular distance was ~2.10-2.25 μm, similar to SL, in the healthy heart in vivo. Therefore, the present high-precision visualization method for the Z-disks or the T-tubules is beneficial to unveiling the mechanisms of myocyte contraction in health and disease in vivo.