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Interstitial lung disease (ILD) is a life-threatening pathological condition that causes respiratory failure and often presents as pulmonary fibrosis. Although it is treated using immunosuppressive and antifibrotic agents, the beneficial effects of these agents remain limited. Thus, the development of new therapeutic strategies for lung fibrosis is crucial. Mesenchymal stem/stromal cells (MSCs) have multilineage differentiation potential; additionally, they have anti-inflammatory and antifibrotic effects as well as the ability to modulate the immune response and modify the microenvironment at the site of engraftment. Numerous adipose-derived MSCs (ASCs) are present in the adipose tissue. Heparin and low-molecular-weight heparin (LMWH) mediate the secretion of several cytokines and growth factors with cell migratory and antifibrotic effects. This study aimed to confirm the therapeutic effect of LMWH-activated ASCs on ILD. Mouse ASCs (mASCs) were cultured in an LMWH-supplemented medium. LMWH significantly increased the number of mASC and enhanced their migratory, anti-inflammatory, and antifibrotic effects. Furthermore, mice with bleomycin-induced pulmonary fibrosis were intravenously administered LMWH-activated mASCs. The relative mRNA expression of inflammation-related genes in ILD lungs was significantly lower in the treatment group than in the pathological model group. Our findings suggest that LMWH-activated mASC administration reduces lung fibrosis.

Exhaled gas analysis is a non-invasive test ideal for continuous monitoring of biological metabolic information. We analyzed the exhaled gas of patients with inflammatory diseases for trace gas components that could serve as biomarkers that enable early detection of inflammatory diseases and assessment of treatment efficacy. Furthermore, we examined the clinical potential of this method. We enrolled 34 patients with inflammatory disease (male/female = 14/20, aged 20–89 years) and 69 healthy participants as controls (male/female ratio = 49/20 aged 30–70 years). Volatile components from exhaled gas were collected and analyzed by a gas chromatography-mass spectrometry system, and the data were examined for gender differences, age, correlation with inflammatory markers (white blood cell count, C-reactive protein, erythrocyte sedimentation rate), and changes in markers before and after treatment. The data was tested for statistical significance through discriminant analysis by Volcano plot, Analysis of variance test, principal component analysis, and cluster analysis comparing healthy and patient groups. There were no significant differences in the trace components of exhaled gas by gender or age. However, we found difference in some components of the exhaled gas between healthy and untreated patients. In addition, after treatment, gas patterns including the patient-specific components changed to a state closer to the inflammation-free status. We identified trace components in the exhaled gas of patients with inflammatory diseases and found that some of these regressed after treatment.

Maxillomandibular advancement surgery is useful for treatment of sleep apnea. However, preoperative analysis and evaluation to facilitate decision-making regarding the direction and distance of maxillomandibular movement has primarily consisted of morphological analysis; physiological function is not evaluated. To improve preoperative prediction, this study used fluid simulation to investigate the characteristics and effects of airway changes associated with maxillomandibular movement. A one-dimensional model with general applicability was thus developed. Actual measurements of flow in patients were used in this fluid simulation, thus achieving an analysis closer to clinical conditions. The simulation results were qualitatively consistent with the actual measurements, which confirmed the usefulness of the simulation. In addition, the results of the one-dimensional model were within the error ranges of the actual measurements. The present results establish a foundation for using accumulating preoperative measurement data for more-precise prediction of postoperative outcomes.

The purpose of this study was to develop a simulation approach for predicting maxillomandibular advancement-induced airway changes using computational fluid dynamics. Eight patients with jaw deformities who underwent maxillomandibular advancement and genioglossus advancement surgery were included in this study. Computed tomography scans and rhinomanometric readings were performed both preoperatively and postoperatively. Computational fluid dynamics models were created, and airflow simulations were performed using computational fluid dynamics software; the preferable number of computational mesh points was at least 10 million cells. The results for the right and left nares, including simulation and postoperative measurements, were qualitatively consistent, and surgery reduced airflow pressure loss. Geometry prediction simulation results were qualitatively consistent with the postoperative stereolithography data and postoperative simulation results. Simulations were performed with either the right or left naris blocked, and the predicted values were similar to those found clinically. In addition, geometry prediction simulation results were qualitatively consistent with the postoperative stereolithography data and postoperative simulation results. These findings suggest that geometry prediction simulation facilitates the preoperative prediction of the postoperative structural outcome.

Japan Agency for Marine-Earth Science and Technology (JAMSTEC) has been disseminated information of data and samples obtained by JAMSTEC's research vessels and submersibles via some databases. The ...

This study aims to achieve high spatial-resolution tandem mass spectrometry (MS/MS) imaging for depicting longitudinal and transverse distribution of drugs in hair, which can provide indispensable information for the proper interpretation of hair test results, including the mechanism of drug incorporation into hair. Two types of hair samples were obtained and analyzed: User's Hair, sampled from a volunteer who took an over-the-counter medicine containing methoxyphenamine (MOP), a nonregulated analogue of methamphetamine; and Soaked Hair, prepared by soaking blank hair in MOP solution. Longitudinal and transverse-sectioning of single hair shafts was accomplished by freeze-sectioning using customized microtomes. Vapor deposition of α-cyano-4-hydroxycinnamic acid provided the finest matrix layer (resolution

In order to investigate the incorporation behavior of drugs into hair in early stage (within 24 h) after intake, time-course changes in drug distribution in black hair were carefully analyzed after a single oral administration of methoxyphenamine (MOP), a non-regulated analog of methamphetamine. Single-hair specimens collected by plucking with the roots intact at appropriate intervals post-intake were each divided into 1-mm segments from the proximal end, and MOP in each segment was determined by a validated liquid chromatography–tandem mass spectrometry procedure. At 10 min after intake, MOP was not detected in any of the segments. MOP became detectable 30 min after intake in the hair bulb (0–1-mm segment from the proximal end) and 1 h after intake in the upper dermis zone (1–2-mm to 4–5-mm segments). The amount of MOP in the hair bulb increased rapidly over 3 h after intake and reached a maximum concentration of ∼100–900 pg/1-mm single hair (11–95 ng/mg) around 3–10 h after intake, whereas that in the upper dermis zone increased at a more gradual pace over 24 h and reached a plateau at ∼30–100 pg/1-mm hair (3–11 ng/mg). These differences can be attributed to the different incorporation mechanisms of the drug. Results from this study can further elucidate the drug incorporation mechanism, which is crucial for accurately interpreting results in hair analyses. Our findings also suggest that hair drug analysis with special attention to the hair root can serve as a useful complementary approach to urine- and blood-based testing in the field of forensic toxicology.

Introduction The aim of this study was to assess the contribution of a large torus mandibularis to induction of obstructive sleep apnea (OSA) due to upper-airway obstruction and to examine the efficacy of torus mandibularis resection in expanding the upper airway. Methods The Japanese version of the Epworth Sleepiness Scale (JESS), unattended portable sleep monitoring and computed tomography (CT) were performed before and at 18 months after surgery. Result The JESS score changed from 6 to 4. The lowest SpO 2 and the percentage of time during which the SpO 2 was 3 to 6,166.58 mm 3 at the level of the posterior soft palate, and from 8,802.93 mm 3 to 12,444.68 mm 3 at the level of the posterior tongue. The volume enlargement ratio evident on 3D image reconstruction was 92.8% at the level of the posterior soft palate and 143.37% at the level of the posterior tongue. The distance from the apex linguae to the anterosuperior point of the hyoid changed from 60 mm to 65 mm, and the distance from the SN plane to the superior point of the dorsum linguae changed from 53 mm to 47 mm. Conclusion Resection of a large torus mandibularis results in expansion of the upper- airway and may reduce airway obstruction during sleep.

The thymus, the organ that is the most sensitive to stress, presents acute involution as a result of exposure to strong stress in childhood. Thymic involution is thus often considered evidence of child abuse/neglect in forensic autopsies. A portion of the thymic epithelial cells express leptin receptor, and leptin showed a thymo-protective function against stress-induced thymic involution in an animal model. Leptin receptor-expressing thymic epithelial cells (LR-TECs) may play a key role in the thymic remodeling provoked by a stressful environment. Here, we sought to clarify the changes of histopathological findings and human LR-TECs in stressful environment. We examined human thymus specimens obtained from 40 forensic autopsy cases (26 male, 14 female; age 21 to 3221 days). We divided the cases into stressor-positive (SP, n = 29) and stressor-negative (SN, n = 11) groups. Cases were classified according to the histological classification of thymic involution and investigated by leptin receptor immunostaining. The results revealed that (1) the SP group showed obvious histological thymic involution (p

育成すべき資質・能力の中核「深く考える（思考力）」の構成要素として，「論理的思考力」及び「メタ認知・学び方の学び」が挙げられている。これを受け，これまでも重視されてきた論理的思考をさらに自覚的なものにすること，つまり「論理的思考の方略化」を図ることが必要であり，そのための学習デザインを検討することが求められると考える。本研究は，国語科の説明的文章を教材とした学習指導において，アクティブ・ラーニングを視点として学習デザインの要件を措定，実践を通して具体化し，その有効性を検証するものである。小学校第５学年児童を対象に実践を行い，論理的思考を促す課題に取り組む学習場面において，学習者の行為である「認知プロセスの外化」の様相から「論理的思考の方略化」の実際を分析・考察した。その結果，「論理的思考の方略化」を図る学習デザインの要件として，探究的な課題（単元レベル）と論理的思考を促す課題（１時間レベル）の設定，対話の重視，パターンや原理を含む教材の選択，思考ツールとしての教具の工夫，学習の振り返りの取り組みの６点が有効であることを明らかにした。

This study evaluated the effect of maxillary advancement surgery on the size of the pharyngeal airway space (PAS). Lateral cephalometric radiographs were collected for 90 patients (29 men and 61 women; average age, 27.2 ± 8.1 years) before (T1) and 1 year after (T2) maxillary advancement surgery. Horizontal and vertical changes in the maxilla and PAS were measured and classified by distance. The maxilla was advanced horizontally by 2.9 ± 1.7 mm and vertically by 2.7 ± 1.4 mm. Upward maxillary movement of ≥4 mm significantly increased PAS (mean change in PAS, 2.6 mm), and upward maxillary movement significantly decreased the posterior nasal spine to the P-point. Only patients with vertical advancement ≥4 mm and horizontal advancement of 3 mm had significant increases in all three PAS parameters. Although forward maxillary movement is believed to have a large effect on PAS, it is suggest that upward vertical movement is more effective for improving PAS. Both the extent and direction of maxillar movement should be considered. Future studies should use cone-beam computed tomography to evaluate the effect of axial direction and differences in PAS.

In order to investigate the influence of pigmentation on the incorporation of drugs into hair, time-course changes in drug distribution along non-pigmented (white) hairs as well as pigmented (black) hairs plucked from the same subject was observed following single administrations of two basic drugs with different properties, zolpidem and methoxyphenamine. These drugs in 1-mm sections of single hair specimens were each determined by a liquid chromatography–tandem mass spectrometric procedure. During the early stage (12–36 h) after intake, for black hairs, both drugs were detected over the entire area of hair root (4–5 mm in length), in which notable concentration of these drugs in the hair bulb (0–1-mm segment from the bottom of hair root, Region 1) and lower concentrations in the upper dermis zone (1–2-mm to 3–4-mm or to 4–5-mm segments, Region 2) were commonly observed. Meanwhile, for white hairs, high drug concentrations in Region 1 as detected in black hairs were not observed although only small amounts of these drugs were detected over Region 2. Subsequent time-course changes in the concentration of drugs in hair demonstrated that the drugs once incorporated into white hair via Region 2 decreased gradually over the period from 24 h to 35 days after intake, but those of black hairs remained almost unchanged. These findings revealed here suggest that hair pigments have two important roles in the distribution of drugs: (1) incorporation of drugs into hair via Region 1, and (2) retention of already incorporated drugs in the hair tissue. These findings would be useful for discussing individual drug-use history based on hair analysis in the forensic fields.

Vestimentiferan tubeworms acquire their symbionts through horizontal transmission from the surrounding environment. In the present study, we constructed a 16S rRNA gene clone library to investigate the phylogenetic relationship between diverse microbes in the sediment and symbiotic bacteria in the trophosome of the tubeworm, Lamellibrachia satsuma, from Kagoshima Bay, Japan. Two symbiotic bacterial phylotypes belonging to the classes γ- and e-Proteobacteria were found from this tubeworm trophosome. They were very closely related to the symbionts of several other marine invertebrates. The most predominant bacteria in the sediment were e-Proteobacteria. A broad diversity of bacteria belonged to non-proteobacterial phyla such as Planctomycetes, Acidobacteria, and Chloroflexi was observed. The presence of sulfur oxidizers (i.e., e-Proteobacteria and γ-Proteobacteria) and sulfur reducers (i.e., δ-Proteobacteria) may play a significant role in the sulfur cycle in these habitats and provide multiple sources of nutrition to the cold-seep communities. Closely related clones of e-Proteobacteria symbiont in the species level and of γ-Proteobacteria symbiont in the genus level were found in the surrounding sediment. The similarity of symbiont clones of L. satsuma with other symbionts and free-living bacteria suggests the possibility of opportunistic symbiosis in e-Proteobacteria and the co-evolution of γ-Proteobacteria having occurred after symbiosis with the tubeworms.

Reactivation of latent infection is considered to be the main mechanism underlying the development of Pneumocystis pneumonia in immunosuppressed patients. We retrospectively assessed the effects of prophylactic administration of sulfamethoxazole-trimethoprim on the development of P. pneumonia and airway colonization with P. jirovecii in patients undergoing examinations to diagnose or rule out P. pneumonia. Polymerase chain reaction was performed to detect P. jirovecii in bronchoalveolar lavage fluid or sputum of 60 consecutive patients between 2004 and 2012. No patients who received the prophylactic administration of sulfamethoxazole-trimethoprim (n = 10) developed P. pneumonia or demonstrated airway colonization with P. jirovecii, and none of the patients who developed P. pneumonia (n = 11) or showed colonization (n = 9) had received prophylactic treatment. Furthermore, 20 (40%) of 50 patients without prophylactic treatment showed positive results on the P. jirovecii DNA polymerase chain reaction, but all 10 patients who had prophylactic treatment showed negative results (Fisher's exact test, P = 0.02). Therefore, the prophylactic administration of sulfamethoxazole-trimethoprim has potential to be effective in preventing P. pneumonia as well as eliminating airway colonization with P. jirovecii. Further studies targeting large cohorts of patients with a variety of underlying diseases are required to develop recommendations regarding the prophylactic administration of sulfamethoxazole-trimethoprim.

Many cell types maintain their function short-term after death. Stem cells isolated from postmortem tissues have been successfully applied in transplantation studies. However, stem cell viability and stemness are reported to decline with increased time after death. Although postmortem stem cells may be useful for regenerative therapy and forensic diagnostics, their characteristic remain to be better understood. Adipose-derived mesenchymal stromal/stem cells (ASCs) have the capacity to differentiate through several cell lineages and are able to survive in an ischemic environment for a prolonged time. This study aimed to confirm whether human postmortem ASCs can be collected and culture-expanded from cadavers. Axilla subcutaneous adipose tissues were harvested during forensic autopsy and enzymatically digested to obtain a heterogeneous cell mixture, including the ASCs population. The mixture was seeded onto collagen-coated cell culture dishes and spindle-shaped adhesive and proliferative ASCs were confirmed. Senescent cells were also present, visualized as large and flattened cells. When maintained in a cool environment, ASCs were able to survive in the postmortem tissues for up to 7 days after death. We conclude that postmortem ASCs can be readily isolated and culture-expanded from adipose tissues.

For forensic pathologists, the most important practice is clarifying the “cause of death” of victims at autopsy. But it is sometimes difficult to determine the cause of death or forensic important matters (e.g., postmortem interval (PMI)) by only gross dissection and microscopic examinations.

Aims Serotonin syndrome (SS) is an adverse outcome of selective serotonin reuptake inhibitors, though its mechanism is not understood and there is no specific clinical biomarker. In this article, metabolic profiles of the SS model rats and causes of metabolome disruption were investigated. Main methods Gas chromatography-tandem mass spectrometry (GC/MS/MS)-based metabolomics, clinical biomarker measurements and qRT-PCR analysis for UCP-3 in skeletal muscles were performed. Key findings Metabolome analysis demonstrated that 55, 22, 49 and 41 of those were significantly altered in plasma, liver, gastrocnemius muscle, and trapezius, respectively. In particular, lactic acid significantly accumulated in the gastrocnemius muscle of the model, while the branched chain amino acids were not consumed in the trapezius, suggesting site differences in abnormal muscular contractions in the model. This result was supported by UCP-3 expression analysis. Alteration of the urea cycle was observed in the liver of the model, attributed mainly to catabolism of proteins and/or amino acids from excess skeletal muscle activity, which was supported by plasma BUN: BUN levels in the model were significantly higher than those in the control. In contrast, almost all metabolites including amino acids and TCA-cycle intermediates significantly increased in plasma of the model, suggesting these were not consumed in some parts of the muscle due to acceleration of anaerobic respiration. Significance Metabolic profiling revealed that abnormal muscular contractions occurred in specific skeletal muscles and enhanced energy production by up-regulation of anaerobic respiration, followed by excess expression of UCP-3, which contributes to the hyper-thermogenesis observed in the SS model.

1-Phenyl-2-(pyrrolidin-1-yl)octan-1-one (PV9) and 16 metabolites, including diastereomers and conjugates, were identified or tentatively detected in human urine by gas chromatography–mass spectrometry and liquid chromatography–high-resolution tandem mass spectrometry. These urinary metabolites indicated that the metabolic pathways of PV9 include: (1) the reduction of ketone groups to their corresponding alcohols; (2) oxidation of the pyrrolidine ring to the corresponding pyrrolidone; (3) aliphatic oxidation of the terminal carbon atom to the corresponding carboxylate form, possibly through an alcohol intermediate (not detected); and (4) hydroxylation at the penultimate carbon atom to the corresponding alcohols followed by further oxidation to ketones, and combinations of these steps. In addition, results from the quantitative analyses of five phase-I metabolites using newly synthesized authentic standards suggested that the main metabolic pathway includes the aliphatic oxidation of terminal and/or penultimate carbons. Human metabolism of PV9 differed significantly from those of α-pyrrolidinovalerophenone and α-pyrrolidinobutiophenone, suggesting that the main metabolic pathways of α-pyrrolidinophenones significantly change depending on the alkyl chain length of the parent molecule.

Estimation of postmortem interval (PMI) is an important goal in judicial autopsy. Although many approaches can estimate PMI through physical findings and biochemical tests, accurate PMI calculation by these conventional methods remains difficult because PMI is readily affected by surrounding conditions, such as ambient temperature and humidity. In this study, Sprague-Dawley (SD) rats (10 weeks) were sacrificed by suffocation, and blood was collected by dissection at various time intervals (0, 3, 6, 12, 24, and 48 h; n = 6) after death. A total of 70 endogenous metabolites were detected in plasma by gas chromatography-tandem mass spectrometry (GC-MS/MS). Each time group was separated from each other on the principal component analysis (PCA) score plot, suggesting that the various endogenous metabolites changed with time after death. To prepare a prediction model of a PMI, a partial least squares (or projection to latent structure, PLS) regression model was constructed using the levels of significantly different metabolites determined by variable importance in the projection (VIP) score and the Kruskal-Wallis test (P < 0.05). Because the constructed PLS regression model could successfully predict each PMI, this model was validated with another validation set (n = 3). In conclusion, plasma metabolic profiling demonstrated its ability to successfully estimate PMI under a certain condition. This result can be considered to be the first step for using the metabolomics method in future forensic casework.

Arrhythmogenic right ventricular cardiomyopathy (ARVC) results in an increased risk of sudden death. We sought mutations of desmoglein-2 (DSG2), desmoplakin (DSP), and plakophilin-2 (PKP2) in 15 cases of sudden death whose causes of death could not be determined at autopsy. In three victims, mutations were identified in DSP. Two of these mutations were novel; one had previously been reported in a patient with ARVC that had been diagnosed clinically. Histological findings were not typical of ARVC; however, it was notable that these mutations were present in three of 15 cases, a relatively high proportion. The causal relationship between the mutations and ARVC is unclear, but the mutations might have been associated with faulty desmosomal proteins resulting in fatal arrhythmia. Combining information gathered by the traditional means of gross and histological examination with postmortem genetic analysis of young victims would assist in identifying their cause of death.

Little information is available on the amounts of hypnotics incorporated into hair after a single administration and about effective analytical procedures to document cases like drug-facilitated sexual assaults. To obtain basic information, single-hair specimens from a volunteer who took a single dose of 10-mg zolpidem (ZP) were analyzed by a newly established liquid chromatography–tandem mass spectrometry procedure, using a one-pot pulverization extraction method. The detection limit of ZP was 50 fg/2-cm single hair, and ZP in each segment was determined for the single black hair specimens (n = 15). ZP was detectable in 14 hairs (positive for all the proximal 0–2 cm segments, negative for all 2–4 cm segments), but was not detected in a single hair (probably in the telogen stage). The amounts of ZP detected in each positive 2-cm segment of single hair ranged from 27 to 63 pg (average 43 pg). The estimated total incorporation of ZP in the scalp hair (black hair ~110,000 strands) was about 4.7 μg, which corresponds to about 0.06 % of the single 10-mg dose (8.03 mg as free ZP). In addition, the direct detection of single-dose ZP incorporated in hair and its imaging were successfully achieved by matrix-assisted laser desorption ionization-mass spectrometry. It is suggested that a combination of these methodologies will provide the highest-level evidence to document such exposure to a hypnotic drug.

In Japan, there are increasing reports of death by poisoning following butane abuse. To determine the specific cause of death in such cases, it is important to confirm the presence of fuel gas components in the body, although careful analysis is required because of their volatile properties. In most reported cases, the subject died suddenly during or immediately after butane aspiration. Thus, the butane concentration in the samples from the deceased should be relatively high. Herein, we present a case of an 18-year-old man found with cardiopulmonary arrest, who then exhibited hypoxic encephalopathy for 16days in a hospital. At autopsy, we detected hypoxic encephalopathy, pneumonia, and ischemia-reperfusion injury of the myocardium, while the cause of cardiac arrest remained unclear. Toxicological analysis was then performed for fuel gas components in several specimens collected at autopsy. Results showed that n-butane and isobutane were detected in the adipose tissue at 16days after inhalation, indicating a role of butane gas inhalation as the cause of death. These data suggest that adipose tissue may be the most appropriate analysis sample to be collected at postmortem in cases where involvement of volatile and fat-soluble gas inhalation is suspected.