Temperature Dependence of the Beating Frequency of hiPSC-CMs Using a MEMS Force Sensor.

It is expected that human iPS cell-derived cardiomyocytes (hiPSC-CMs) can be used to treat serious heart diseases. However, the properties and functions of human adult cardiomyocytes and hiPSC-CMs, including cell maturation, differ. In this study, we focused on the temperature dependence of hiPSC-CMs by integrating the temperature regulation system into our sensor platform, which can directly and quantitatively measure their mechanical motion. We measured the beating frequency of hiPSC-CMs at different environmental temperatures and found that the beating frequency increased as the temperature increased. Although the rate at which the beating frequency increased with temperature varied, the temperature at which the beating stopped was relatively stable at approximately 20 °C. The stopping of beating at this temperature was stable, even in immature hiPSC-CMs, and was considered to be a primitive property of cardiomyocytes.

Muscular Hemorrhages Around the Scapula Provide Insight on the Manner of Asphyxia: A Preliminary Study.

ABSTRACT: Muscular hemorrhages around the scapula that are caused by upper extremity motion are rarely reported in cases other than drowning. We examined differences in the frequency of muscular hemorrhages around the scapula and stratified them by the cause of death. Muscular hemorrhages were mostly noted in cases of drowning (118/185, 63.8%), followed by cases of asphyxia (23/44, 52.3%). In addition, muscular hemorrhages around the scapula were found in cases of ligature strangulation (8/11, 72.7%), manual strangulation (2/3, 66.7%), choking (11/14, 78.6%), and traumatic asphyxia (2/2, 100%). Muscular hemorrhages were not found in cases of hanging or environmental suffocation, possibly because of the short time interval preceding the loss of consciousness. The distribution of muscular hemorrhages was similar in choking and drowning cases. Muscular hemorrhages due to strangulation were mainly unilateral, whereas those due to choking and drowning were mainly bilateral. During all types of asphyxia, muscular hemorrhages around the scapula were considered to be the result of conscious, active upper extremity motion. Because the frequency and laterality of muscular hemorrhages around the scapula differed based on the type of asphyxia, our findings provide insights into the manner of asphyxia that could be used to aid in the identification of homicidal hangings.

A Case of Personal Identification Via Postmortem Computed Tomography Confirmation of Aphakic Eyes.

Postmortem computed tomography (PMCT) is used commonly in forensic medicine. Postmortem computed tomography of the head provides information on the eyes and orbits and may reveal intraocular abnormalities. We present a case in which the identity of a deceased man was confirmed by the presence of aphakic eyes detected on PMCT. A decomposed body was found in a house and suspected to be a man in his 40s who lived alone. Autopsy identified pontine hemorrhage as the cause of death. Although signs of previous dental treatment were evident, the storage period of the householder's dental records had expired. He had no family members to provide DNA for a comparative DNA analysis. Postmortem computed tomography before autopsy revealed aphakia (ie, the absence of eye lenses) suggestive of previous ocular surgery. His medical records revealed that he had undergone ocular surgery for secondary glaucoma due to uveitis, which had resulted in artificial aphakia. Peripheral iridectomy scars fully matched the findings in his medical records, thus identifying the body. The unusual finding of artificial aphakia in a man in his 40s facilitated the personal identification in this case. In conclusion, PMCT provides additional information, and the correct interpretation of intraocular PMCT findings by forensic pathologists is crucial.

Effect of drug-coated balloon angioplasty on in-stent restenotic coronary lesions analyzed with optical coherence tomography and serial coronary artery angioscopy.

Drug-coated balloon angioplasty (DCBA) has been recognized for its utility in preventing in-stent re-restenosis (ISR); however, imaging of the neointima immediately after treatment and during follow-up has only been described in a few case reports. This study aimed to determine the efficacy and mechanism of the DCBA using imaging studies both immediately after the DCBA and during the follow-up period. We enrolled 15 consecutive patients who underwent DCBA for in-stent restenosis (ISR). The in-stent neointimal volume was evaluated using optical coherence tomography (OCT), and the in-stent yellow grade was assessed using coronary angioscopy (CAS) immediately after DCBA and during the median follow-up period of 9 (8-15) months. The neointimal volume was significantly reduced from 77.1 ± 36.2 mm3 at baseline to 60.2 ± 23.9 mm3 immediately after DCBA (p = 0.0012 vs. baseline) and to 46.7 ± 21.9 mm3 during the follow-up (p = 0.0002 vs. post DCBA). The yellow grade of the residual plaques at the ISR lesion, which indicated plaque vulnerability, was significantly decreased in the follow-up CAG (from baseline: 1.79 ± 1.03, during the follow-up: 0.76 ± 0.82; p < 0.0001). These data suggest that DCBA may inhibit neointimal formation and provide angioscopic intimal stabilization for ISR lesions.

Effect of Aggressive lipid-lowering treatment with Rosuvastatin on vascular endoTHelium function: evaluation of vascular endothelium function (EARTH study).

Vascular endothelial dysfunction plays an important role in the process of atherosclerosis up to the final stage of plaque rupture. Vascular endothelial dysfunction is reversible, and can be recovered by medications and life-style changes. Improvement in endothelial function may reduce cardiovascular events and improve long-term prognosis. A total of 50 patients with stable angina and dyslipidemia were enrolled, including patients who had not received prior treatment with statins and had serum LDL-C levels ≥ 100 mg/dL, and patients who had previously received statin treatment. All agreed to register regardless of their LDL-C level. Rosuvastatin was initially administered at a dose of 2.5 mg and appropriately titrated up to the maximum dose of 20 mg or until LDL-C levels lower than 80 mg/dL were achieved, for 24 weeks. Endothelial function was assessed by the reactive hyperemia peripheral arterial tonometry (RH-PAT) index in the radial artery by Endo-PAT® 2000 (Endo-PAT®2000, software version 3.0.4, Itamar Medical Ltd., Caesarea, Israel). RH-PAT data were digitally analyzed online by Endo-PAT®2000 at baseline and at 24 weeks. LDL-C and MDA-LDL-C decreased from 112.6 ± 23.3 to 85.5 ± 20.2 mg/dL and from 135.1 ± 36.4 to 113.9 ± 23.5 mg/dL respectively (p < 0.0001). However, HDL-C, hs-CRP and TG did not change significantly after treatment. RH-PAT index levels significantly improved, from 1.60 ± 0.31 to 1.77 ± 0.57 (p = 0.04) after treatment, and the percent change of the RH-PAT index was 12.8 ± 36.9%. Results of multivariate analysis show that serum LDL-C levels over 24 weeks did not act as a predictor of improvement of the RH-PAT index. However, HbA1c at baseline was an independent predictor which influenced the 24-week RH-PAT index level. The RH-PAT index of patients with high HbA1c at baseline did not improve after administration of rosuvastatin but it did improve in patients with low HbA1c at baseline. Aggressive lowering of LDL-C with rosuvastatin significantly improved the RH-PAT index, suggesting that it may improve endothelial function in patients with coronary artery disease.Clinical Trial Registration No: UMIN-CTR, UMIN000010040.

A case of carbon monoxide poisoning before a vehicle fire: Availability of volatile hydrocarbons analysis.

Benzene is one of volatile hydrocarbons contained in fire smoke, and the concentrations in the blood are known to be positively correlated with that of carbon monoxide-hemoglobin (CO-Hb) in fire-related deaths. In this report, we present a vehicle fire case in which CO and benzene concentration is atypically un-correlated. The car driven by the vehicle dweller ran into an oncoming lane at high speed, hitting a traffic signal pole. A vehicle fire started when the rescuer opened the car door. A burned body and briquette stove were found when the fire was extinguished. An autopsy revealed a small amount of soot deposit in the airways. The CO-Hb concentration in the heart blood was 63%. Volatile hydrocarbon analysis of the blood was performed; compared with the CO-Hb concentration, the benzene concentration was significantly lower than expected. High CO-Hb concentration without a hydrocarbon component indicated that the deceased inhaled CO that was not related to fire smoke. Thus, we concluded that the cause of death was CO poisoning caused by the briquette stove before the vehicle fire started. Comparing volatile hydrocarbon concentrations with CO-Hb concentrations could provide more information about the circumstances surrounding a vehicle fire-related death.

Toxicologic pathological mechanism of acute lung injury induced by oral administration of benzalkonium chloride in mice.

Benzalkonium chloride (BAC) intoxication causes fatal lung injuries, such as acute lung injury (ALI) and acute respiratory distress syndrome (ARDS). However, the pathogenesis of ALI/ARDS induced by BAC ingestion is poorly understood. This study aimed to clarify the mechanism of lung toxicity after BAC ingestion in a mouse model. BAC was orally administered to C57BL/6 mice at doses of 100, 250, and 1250 mg/kg. After administration, BAC concentrations in the blood and lungs were evaluated via liquid chromatography with tandem mass spectrometry. Lung tissue injury was evaluated via histological and protein analyses. Blood and lung BAC concentration levels after oral administration increased in a dose-dependent manner, with the concentrations directly proportional to the dose administered. The severity of lung injury worsened over time after the oral administration of 1250 mg/kg BAC. An increase in the terminal transferase dUTP nick end labeling-positive cells and cleaved caspase-3 levels was observed in the lungs after 1250 mg/kg BAC administration. In addition, increased cleaved caspase-9 levels and mitochondrial cytochrome c release into the cytosol were observed. These results suggest that lung tissue injury with excessive apoptosis contributes to BAC-induced ALI development and exacerbation. Our findings provide useful information for developing an effective treatment for ALI/ARDS induced by BAC ingestion.

Detection of helium in a fire victim: A case report.

We report here detection of helium in specimens derived from a burn autopsy case. A male was found in a burnt bedroom. Part of a heat-denatured plastic bag, sealing tape, and flexible tubing remained on his head and neck. In addition, five helium tanks were found near him. His history in conjunction with the discovery conditions suggested a suicide attempt by inhalation of helium. The body had extensive first to fourth degree burns caused by heat. A small amount of soot was deposited in the respiratory tract. Except for the thermal burns, no other injuries were found. Toxicologically, the blood carboxyhemoglobin saturation levels were less than 6%, while combustion-derived volatile hydrocarbons such as benzene or toluene were detected in the blood. In addition, tracheal gas, gastric gas, headspace gas of lung tissue, brain, and heart blood were collected during autopsy for detection of helium. Analysis was performed using headspace gas chromatography with a thermal conductivity detector. Helium was detected in all of the samples tested. Etizolam at a low limit of therapeutic concentration or less was detected in the blood. Neither ethanol nor other drugs of abuse were detected in his blood or urine. Autopsy findings and experiments suggest that the victim inhaled helium and was still alive when a fire broke out. The cause of his death was diagnosed as death from fire and flames. The present result suggests that helium may remain in a burned body and that investigation of helium in cases of fire-related deaths is informative for determination of the cause of death or confirmation of the ante mortem involvement of helium.

Back hemorrhage in bath-related deaths: Insights into the mechanism of bath-related deaths.

Forensic pathologists often encounter difficulties while diagnosing bath-related deaths owing to a lack of specific findings for determining the cause. This study reports the characteristic findings noted during autopsy associated with bath-related deaths. The subjects included individuals found deceased in water-filled bathtubs. Originally, a "bathtub death" was defined as a case of drowning in a bathtub without a known cause despite complete investigation including a complete autopsy and examination of the death scene. We also excluded cases with ethanol and/or high drug concentrations in the blood from "bathtub death" cases. Muscular hemorrhages around the scapula were the most common in cases of accidental drowning (72.7%). The frequency of muscular hemorrhages around the scapula was low in cases involving bathtub deaths (11.1%) and natural disease. Subcutaneous hemorrhages in the back were the most frequent in cases of bathtub death (88.9%), followed by accidental drowning (81.8%). The frequency of subcutaneous hemorrhages in the back was low in cases involving suicide and natural disease. The presence of subcutaneous hemorrhages in the back without muscular hemorrhages around the scapula were strongly associated with bathtub deaths (Fisher's exact test, P < 0.0001; sensitivity, 0.84; specificity, 0.91). Subcutaneous hemorrhages were the most frequent in the waist (68.8%), followed by the upper back (50%). Subcutaneous hemorrhages in the back without muscular hemorrhages around the scapula indicated that the deceased had hit their back on falling from a standing position after experiencing disturbances of consciousness. We found the occurrence of orthostatic hypotension while getting out of the bathtub to be the most likely cause for these autopsy findings. Subcutaneous hemorrhages in the back without muscular hemorrhages around the scapula is a characteristic finding of "bathtub deaths". Although this was a preliminary study, the presence of this finding may aid in clarifying the mechanism of bath-related deaths.

Muscular hemorrhages around the scapula resulting from excessive upper extremity motion in cases of fatal drowning: autopsy findings for insights on manner of death.

Muscular hemorrhages around the scapula are frequently found in fatal drowning cases without injuries in the upper back at time of autopsy. The present study investigated the frequency of muscular hemorrhages around the scapula and determined the mechanism responsible for this phenomenon. Muscular hemorrhages around the scapula were found in 104 of 164 (63.4%) bodies assessed. Hemorrhage in the infraspinatus muscle was most common, followed by the supraspinatus muscle. These muscular hemorrhages were not associated with upper extremity injuries and were most frequently found in accidental drowning cases (78.7%), followed by cases of suicide (56.0%), and in natural disease prior to drowning (19.0%). We examined muscular hemorrhage around the scapula and cervical injuries that restricted the active motion of upper extremities in drowning cases. The vertebral level of cervical injuries were related to muscular hemorrhages around the scapula. These results suggested that muscular hemorrhages around the scapula were not a result of agonal convulsions but were caused by consciously active excessive motion of the upper extremities while drowning. Investigations of the muscular hemorrhages around the scapula can provide insights towards the manner of fatal drowning.

Protective role for CD1d-reactive invariant natural killer T cells in cauterization-induced corneal inflammation.

PURPOSE: Corneal inflammation can be induced by various stimuli, such as chemical burns, trauma, and acute bacterial infection, and directly impairs visual acuity. Natural killer T (NKT) cells belong to a specialized population of leukocytes that coexpress the T-cell receptor and NK markers. This study examined the role of CD1d-reactive invariant NKT cells in cauterization-induced acute corneal inflammation. METHODS: The corneas of CD1d-knockout (KO) mice and Jalpha18-KO mice (both of which are NKT cell deficient) and control mice were cauterized with silver nitrate. Corneal edema and opacity were examined, and the phenotypes of the corneal-infiltrating cells were analyzed histologically at 24 hours and by flow cytometry at 96 hours. Reverse transcription-polymerase chain reaction (RT-PCR) was used to determine the expression of vascular endothelial growth factor (VEGF), interferon (IFN)gamma, and tumor necrosis factor (TNF)alpha in the cauterized corneas. RESULTS: The CD1d-KO and Jalpha18-KO mice had significantly greater levels of corneal edema and opacity than did the control mice. Although the number of infiltrating cells was not significantly different at 96 hours, both groups of NKT cell-deficient mice demonstrated increased early neutrophil accumulation at 24 hours and early expression of VEGF, IFNgamma, and TNFalpha. There was no difference in the level of VEGF-induced corneal neovascularization. CONCLUSIONS: NKT cells appear to regulate the early accumulation of neutrophils, protect the cornea from excessive inflammation, and maintain corneal clarity. However, in this study, they did not affect the corneal revascularization process induced by VEGF.

A novel beta-myosin heavy chain gene mutation, p.Met531Arg, identified in isolated left ventricular non-compaction in humans, results in left ventricular hypertrophy that progresses to dilation in a mouse model.

Mutations in the betaMHC (beta-myosin heavy chain), a sarcomeric protein are responsible for hypertrophic and dilated cardiomyopathy. However, the mechanisms whereby distinct mutations in the betaMHC gene cause two kinds of cardiomyopathy are still unclear. In the present study we report a novel betaMHC mutation found in a patient with isolated LVNC [LV (left ventricular) non-compaction] and the phenotype of a mouse mutant model carrying the same mutation. To find the mutation responsible, we searched for genomic mutations in 99 unrelated probands with dilated cardiomyopathy and five probands with isolated LVNC, and identified a p.Met531Arg mutation in betaMHC in a 13-year-old girl with isolated LVNC. Next, we generated six lines of transgenic mice carrying a p.Met532Arg mutant alphaMHC gene, which was identical with the p.Met531Arg mutation in the human betaMHC. Among these, two lines with strong expression of the mutant alphaMHC gene were chosen for further studies. Although they did not exhibit the features characteristic of LVNC, approx. 50% and 70% of transgenic mice in each line displayed LVH (LV hypertrophy) by 2-3 months of age. Furthermore, LVD (LV dilation) developed in approx. 25% of transgenic mice by 18 months of age, demonstrating biphasic changes in LV wall thickness. The present study supports the idea that common mechanisms may be involved in LVH and LVD. The novel mouse model generated can provide important information for the understanding of the pathological processes and aetiology of cardiac dilation in humans.

Injury patterns of fatal bear attacks in Japan: A description of seven cases.

In recent years, the number of bear attacks has risen in Akita prefecture, Japan. Here, we present the injury patterns of seven fatal bear attacks, and discuss the implication of these findings. We included five cases of Asiatic black bear attacks and two cases of Ezo brown bear attacks. In all cases, the injuries, 2-5 parallel linear lacerations with severe hemorrhaging and decollement, were mainly located on the upper body. These injuries were thought to be fatal as, upon a first encounter, bears often stand and first attack the victim's head and face using their claws. Four lacerations were located at the vertex of the trapezoid in all cases, without severe hemorrhaging on the neck, extremities, or around the antemortem injuries. These injuries were thought to be bite marks incurred by the bears' four large canines, mainly occurring postmortem during the process of predation. These findings differed from those of fatal biting around the neck by other animals such as lions, mountain lions, or large-sized dogs. Further, laceration with avulsion of the skin was found in the inguinal region, without severe hemorrhaging. In some cases, the intestine had been removed from the inguinal injury for predation. In conclusion, the injuries of bear attacks are different from those caused by other animals, owing to the characteristics of bears. By investigating the injuries caused by fatal bear attacks, we can better understand the patterns of such injuries. Especially, the diameter between the canines is sometimes useful to estimate the size and the number of offending bears.

Visualization of old bruises in children: Use of violet light to record long-term bruises.

When a child consultation center evaluates abused children, evidence of the abuse must be documented. Children who have suffered physical abuse often have bruises, which cannot be confirmed by visualization when the bruise has healed. In forensics, a method to confirm an old bruise by ultraviolet radiation is possible. However, long-term ultraviolet exposure can be harmful. In this study, we observed 15 bruises on healthy volunteer children using four kinds of light sources, including ultraviolet radiation, over time. The violet light source enabled observation of a bruise for a longer time than ultraviolet radiation. Although other studies have shown that violet light is superior for enhancing bruises, this is the first report to evaluate data concerning the ability to identify bruises over time in living children. These results might help to identify evidence of past violence suffered by children.

Development of a new mouse model of branch retinal vein occlusion and retinal neovascularization.

PURPOSE: Retinal neovascularization (NV) is associated with various disorders, such as retinal vein occlusion, diabetic retinopathy, and retinopathy of prematurity, and often causes severe loss of vision. To determine the mechanism of retinal NV and develop new therapy, we developed a mouse model using a photodynamic method. METHODS: C57BL/6 mice were injected with rose bengal via the tail vein, and then selected venous points were photocoagulated. RESULTS: All eyes demonstrated venous occlusion on day 1, and capillary nonperfusion areas were observed until day 3. Twenty of 33 eyes (60.6%) developed retinal NV on day 14, confirmed by fluorescein isothiocyanate-perfused retinal flat-mounts and immunochemical and histopathological analyses. Reverse transcriptase-polymerase chain reaction showed an increase in the expression of vascular endothelial growth factor at the retina on day 7. CONCLUSIONS: Because of the simplicity, low cost, and feasibility of genetic manipulations, our model is believed to represent an advance in investigating molecular mechanisms and establishing therapy for retinal NV.

Case studies in poststroke hemiplegic patients using SEMUL: a passive 2-DOF rehabilitation robot.

Following stroke, rehabilitation exercises paired with medical treatment are important for the recovery and maintenance of upper limb function. We developed a 2-DOF passive haptic system for upper limb rehabilitation and software focusing on reaching exercises, known as the coin collecting program, which provides training and evaluation of motor function, particularly dexterity. Past studies have revealed quantitative, reproducible evaluation indices using this software. In this paper, we compared case studies of post-stroke patients with results from healthy participants, and confirmed the reliability of the evaluation indices in grading or scoring motor skills.

A case of giant saphenous vein graft aneurysm followed serially after coronary artery bypass surgery.

Saphenous vein graft aneurysm (SVGA) is one of the chronic complications after coronary aorta bypass grafting (CABG) and may be caused by atherosclerosis-like phenomena of the vein graft, weakness around the vein valve, rupturing of the suture of the graft anastomosis, or perioperative graft injury. We describe a case of a large, growing saphenous vein graft aneurysm that was followed serially by chest radiography and computed tomography. Eighteen years after CABG, an SVGA (23 × 24 mm) was incidentally detected. The patient was asymptomatic and was followed conservatively. Four years later, coronary computed tomographic angiography showed that the giant aneurysm had grown to 52.1 by 63.8 mm and revealed a second, smaller aneurysm. Finally, the SVG was ultimately resected without bypass via off-pump surgery. Therefore, this case suggested that aggressive treatment that includes surgical intervention should be considered before the aneurysm becomes larger, even if it is asymptomatic.

Cellular events in the normal and inflamed cornea.

PURPOSE: Although normal cornea is an avascular transparent structure, bone marrow (BM)-derived cells exist in situ even in noninflamed conditions. We evaluated constitutive cellular trafficking into the cornea in the naive state and investigated how corneal inflammation may be initiated by various stimuli. METHODS: BM chimeric mice were generated using BM from enhanced green fluorescent protein (eGFP) transgenic mice. Corneas of chimeric mice were carefully studied by fluorescent biomicroscopy until 6 months after transplantation. To analyze initiation of cellular events in corneal immune response, we cauterized the center of the cornea or inoculated IL-1 beta into the corneal micropocket. Cellular events were analyzed by flow cytometry. RESULTS: At 2 weeks after BM transplantation, GFP cells gradually migrated into the cornea from the limbal area and were distributed over the entire cornea at 6 months. In both the cauterization and micropocket assays, infiltration of neutrophils and macrophages occurred on days 2 and 4, respectively. Depletion of neutrophils by anti-Gr-1 Ab significantly reduced corneal edema/opacity induced by cauterization. IL-1 beta-induced angiogenesis was markedly reduced in monocyte chemoattractant protein-1 knockout mice. CONCLUSIONS: BM-derived cells are recruited into the normal cornea and may be essential to maintain corneal clarity. In the inflamed cornea, neutrophils might be responsible for acute corneal edema/opacity and macrophages for corneal angiogenesis and chronic inflammation.

Three cases of suprachoroidal hemorrhage associated with chest compression or asphyxiation and detected using postmortem computed tomography.

We report 3 cases of suprachoroidal hemorrhage (SCH) found to be triggered by increased intrathoracic pressure and detected using postmortem computed tomography (PMCT). Case 1 was a man aged in his 50s who was found dead at a landslide site. The autopsy showed clogging of the upper respiratory tract with soil debris from the landslide. The cause of death was determined to be asphyxia. PMCT showed SCH in both eyes, which was believed to be caused by chest compression or choking on the soil debris from the landslide. Case 2 was a woman aged in her 60s who was found dead in the sea. The autopsy revealed injuries primarily to her chest. We concluded that the cause of death was drowning. PMCT showed SCH in her right eye that was believed to be caused by chest compression. Case 3 was a woman aged in her 80s who was buried in a snowdrift and potentially died from hypothermia. PMCT showed SCH in both eyes, which was considered to be from an increase in intrathoracic pressure that might have been caused by the burial in the snow. Histological findings showed serous retinal detachment associated with retinal pigment epithelium damage due to SCH, which indicated that she was alive for several hours after the onset of SCH. The increase in intrathoracic pressure caused by dyspnea or chest compression was considered responsible for the onset of SCH in all of the present cases. PMCT might assist with the differential diagnosis of traumatic asphyxiation by SCH.