Search

Exosomes are small extracellular vesicles (EVs) found in culture supernatants, blood, and breast milk. The size of these nanocomplexes limits the methods of EV analyses. In this study, nitrobenzoxadiazole (NBD), a fluorophore, conjugated endosome-lysosome imager, GIF-2250 and its derivative, GIF-2276, were evaluated for exosome analyses. A correlation was established between GIF-2250 intensity and protein maker levels in bovine milk exosomes. We found that high-temperature sterilization milk may not contain intact exosomes. For precise analysis, we synthesized GIF-2276, which allows for the covalent attachment of NBD to the Lys residue of exosome proteins, and labeled milk exosomes were separated using a gel filtration system. GIF-2276 showed chromatographic peaks of milk exosomes containing >3 ng protein. The area (quantity) and retention time (size) of the exosome peaks were correlated to biological activity (NO synthesis suppression in RAW264.7 murine macrophages). Heat denaturation of purified milk-derived exosomes disrupted these indicators. Proteome analyses revealed GIF-2276-labeled immunomodulators, such as butyrophilin subfamily 1 member A1 and polymeric immunoglobulin receptor. The immunogenicity and quantity of these factors decreased by heat denaturation. When milk exosomes were purified from market-sourced milk we found that raw and low-temperature sterilization milk samples, contained exosomes (none in high-temperature sterilization milk). These results were also supported by transmission electron microscopy analyses. We also found that GIF-2276 could monitor exosome transportation into HEK293 cells. These results suggested that GIF-2250/2276 may be helpful to evaluate milk exosomes., Competing Interests: Declaration of competing interest H.T. and S.Y. owns 3600 USD of Gifu Exosome Co. LTD. shares. K.F. and Y.M. owns 1800 USD of Gifu Exosome shares. S.Y. is employed by Seki Milk Co. LTD. Others have nothing to declare. GIF-2250 and GIF-2276 have been submitted patents, WO2022163446 and PCT/JP2023/28846., (Copyright © 2024 Elsevier Inc. All rights reserved.)

Background: Latent chronic inflammation has been proposed as a key mediator of multiple derangements in metabolic syndrome (MetS), which are increasingly becoming recognized as risk factors for age-related cognitive decline. However, the question remains whether latent chronic inflammation indeed induces brain inflammation and cognitive decline., Methods: A mouse model of latent chronic inflammation was constructed by a chronic subcutaneous infusion of low dose lipopolysaccharide (LPS) for four weeks. A receptor for advanced glycation end products (RAGE) knockout mouse, a chimeric myeloid cell specific RAGE-deficient mouse established by bone marrow transplantation and a human endogenous secretory RAGE (esRAGE) overexpressing adenovirus system were utilized to examine the role of RAGE in vivo. The cognitive function was examined by a Y-maze test, and the expression level of genes was determined by quantitative RT-PCR, western blot, immunohistochemical staining, or ELISA assays., Results: Latent chronic inflammation induced MetS features in C57BL/6J mice, which were associated with cognitive decline and brain inflammation characterized by microgliosis, monocyte infiltration and endothelial inflammation, without significant changes in circulating cytokines including TNF-α and IL-1β. These changes as well as cognitive impairment were rescued in RAGE knockout mice or chimeric mice lacking RAGE in bone marrow cells. P-selectin glycoprotein ligand-1 (PSGL-1), a critical adhesion molecule, was induced in circulating mononuclear cells in latent chronic inflammation in wild-type but not RAGE knockout mice. These inflammatory changes and cognitive decline induced in the wild-type mice were ameliorated by an adenoviral increase in circulating esRAGE. Meanwhile, chimeric RAGE knockout mice possessing RAGE in myeloid cells were still resistant to cognitive decline and brain inflammation., Conclusions: These findings indicate that RAGE in inflammatory cells is necessary to mediate stimuli of latent chronic inflammation that cause brain inflammation and cognitive decline, potentially by orchestrating monocyte activation via regulation of PSGL-1 expression. Our results also suggest esRAGE-mediated inflammatory regulation as a potential therapeutic option for cognitive dysfunction in MetS with latent chronic inflammation., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Inc. All rights reserved.)

Recent studies have shown a role of inflammation in muscle atrophy and sarcopenia. However, no anti-inflammatory pharmacotherapy has been established for the treatment of sarcopenia. Here, we investigate the potential role of PPARα and its ligands on inflammatory response and PGC-1α gene expression in LPS-treated C2C12 myotubes. Knockdown of PPARα, whose expression was upregulated upon differentiation, augmented IL-6 or TNFα gene expression. Conversely, PPARα overexpression or its activation by ligands suppressed 2-h LPS-induced cytokine expression, with pemafibrate attenuating NF-κB or STAT3 phosphorylation. Of note, reduction of PGC-1α gene expression by LPS treatment for 24 hours was partially reversed by fenofibrate. Our data demonstrate a critical inhibitory role of PPARα in inflammatory response of C2C12 myotubes and suggest a future possibility of PPARα ligands as a candidate for anti-inflammatory therapy against sarcopenia., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Tomohiro Tanaka reports a relationship with Kowa Company Ltd that includes: speaking and lecture fees. All the other authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Inc. All rights reserved.)

Amino acids exist in two chiral forms, namely L and D. Although l-amino acids are predominant in vivo, certain limited circumstances have reported the usage of d-amino acids. d-aspartate (Asp), among them, plays crucial physiological roles in living organisms and is biosynthesized from L-Asp by the enzyme named aspartate racemase (AspRase). D-Asp is known to accumulate in large amounts in the nervous system of cephalopods. To understand the function of D-Asp in nervous system in more detail, it is necessary to elucidate its metabolic pathway; however, AspRase gene has not been identified in cephalopods as in the case of mammals. In this study, we successfully identified a novel gene encoding AspRase from the optic ganglion of Japanese common squid Todarodes pacificus. Our discovery of the squid AspRase challenges the prevailing assumption that AspRases across different animals share similar structures. Surprisingly, the squid AspRase is a unique enzyme that differs significantly from known AspRases, being structurally and phylogenetically related to aspartate aminotransferase (AST) and possessing both AspRase and AST activities. The optimum pH and temperature for AspRase activity using L-Asp as a substrate are approximately 7.0 and 20 °C, respectively. Moreover, we have found that AspRase activity is enhanced in the presence of 2-oxoacids. These findings have far-reaching implications for the understanding of enzymology and suggest that yet-to-be-identified mammalian AspRases may also be phylogenetically related to AST, rather than conventional AspRases. Furthermore, our results provide valuable insights into the evolution of the D-Asp biosynthetic pathway., (Copyright © 2023 Elsevier Inc. All rights reserved.)

Non-human primate (NHP) renal transplantation models are widely used vivo models for researching new immunosuppressive therapies including allograft tolerance strategies. To enroll animals into a tolerance study, an immunosuppressive regimen that efficiently establishes stable renal function in NHPs is needed. Here, we assessed the effect of triple therapy comprising 2.0 mg/kg tacrolimus, mycophenolate mofetil and a steroid and its success rate for achieving stable renal function. In addition, to predict the pathophysiological consequences of withdrawing immunosuppressants, an indispensable process after induction of tolerance, we also assessed changes in the stable renal state maintained by triple therapy after drug withdrawal. Six cynomolgus monkeys were used. The median survival time was >176 days over the dosing period and 45 days after drug withdrawal. The triple therapy successfully induced stable graft function without calcineurin inhibitor nephrotoxicity in three of six recipients, although adopting trough-dependent tacrolimus dose adjustment rather than a preset dose regimen could improve on the present strategy. Further, drug withdrawal led to deterioration of renal function, de novo donor specific antibody production and increased the memory/naïve T cell ratio within two weeks post drug withdrawal. We expect that these findings contribute to establish one of the choices for animal model for evaluating future tolerance therapy for renal transplantation., Competing Interests: Declaration of Competing Interest YN, MM, KN, HF, HS, RM, KH, YH, HK, TM were employees of Astellas Pharma Inc. and Astellas Research Technologies Co., Ltd. during the conduct of the study., (Copyright © 2022 The Authors. Published by Elsevier B.V. All rights reserved.)

Introduction: Complicated skin and soft tissue infections (cSSTIs) and bacteremia caused by Staphylococcus aureus, including methicillin-resistant S. aureus (MRSA), are common causes of infection for children worldwide. Here, the safety and efficacy of daptomycin in Japanese pediatric participants are reported., Methods: This open-label, single-arm phase 2 study (NCT03643952) enrolled Japanese pediatric participants (age 1-17 years) with cSSTI or bacteremia caused by gram-positive cocci. Participants received age-adjusted doses of intravenous daptomycin for 5 to up to 14 days (cSSTI) or 5 to up to 42 days (bacteremia). The primary objective was safety and tolerability; efficacy among participants with infections caused by MRSA was a secondary objective., Results: A total of 18 participants (cSSTI, n = 14; bacteremia, n = 4) were enrolled across 12 study sites in Japan. The most common pathogen was S. aureus (15/18 [83.3%]), including methicillin-susceptible and -resistant isolates. Adverse events (AE) were reported in 42.9% (6/14) of participants with cSSTI and 100% (4/4) of participants with bacteremia. No deaths, serious AEs, discontinuations of study medication due to an AE, or events of clinical interest occurred in the study. In participants with infections caused by MRSA, 87.5% [7/8] achieved favorable clinical response at test of cure (TOC) visit (cSSTI, 85.7% [6/7]; bacteremia, 100% [1/1]). In this population, favorable microbiological response at TOC was achieved by 71.4% (5/7) of participants with cSSTI and 100% (1/1) of participants with bacteremia., Conclusions: Daptomycin was well tolerated, exhibited a favorable safety profile, and was effective for the treatment of cSSTI or bacteremia in Japanese children., (Copyright © 2021. Published by Elsevier Ltd.)

Smoking cessation increases body weight. The underlying mechanisms, however, have not been fully understood. We here report an establishment of a mouse model that exhibits an augmented body weight gain after nicotine withdrawal. High fat diet-fed mice were infused with nicotine for two weeks, and then with vehicle for another two weeks using osmotic minipumps. Body weight increased immediately after nicotine cessation and was significantly higher than that of mice continued on nicotine. Mice switched to vehicle consumed more food than nicotine-continued mice during the first week of cessation, while oxygen consumption was comparable. Elevated expression of orexigenic agouti-related peptide was observed in the hypothalamic appetite center. Pair-feeding experiment revealed that the accelerated weight gain after nicotine withdrawal is explained by enhanced energy intake. As a showcase of an efficacy of pharmacologic intervention, exendin-4 was administered and showed a potent suppression of energy intake and weight gain in mice withdrawn from nicotine. Our current model provides a unique platform for the investigation of the changes of energy regulation after smoking cessation., Competing Interests: Declaration of competing interest Tomohiro Tanaka reports financial support was provided by Japan Society for the Promotion of Science. Daisuke Aotani reports financial support was provided by Japan Society for the Promotion of Science. Takashi Yagi reports financial support was provided by Japan Society for the Promotion of Science. Tingting Guo reports financial support was provided by Japan Society for the Promotion of Science. Tomohiro Tanaka reports financial support was provided by Smoking Research Foundation. Daisuke Aotani reports financial support was provided by Smoking Research Foundation., (Copyright © 2021 Elsevier Inc. All rights reserved.)

Obesity is a growing worldwide public health issue and is associated with a range of comorbidities, including cognitive deficits. The present study investigated synaptic changes in the hippocampus during the development of obesity. The treatment of newborn mice with monosodium-L-glutamate (MSG, 2 mg/g) induced obesity and recognition memory deficits in the novel object recognition (NOR) test at 16-17 weeks, but not at 8-9 weeks. Hippocampal synaptic plasticity, including long-term potentiation (LTP) and long-term depression (LTD), and excitatory synaptic transmission at Schaffer collateral-CA1 (SC-CA1) synapses were compared between MSG-treated mice and age-matched control mice. LTP and fiber volley amplitudes were enhanced in MSG-treated mice at 16-17 weeks, but not at 8-9 weeks. Furthermore, the strength of paired-pulse facilitation (PPF) changed in MSG-treated mice at 16-17 weeks, but not at 8-9 weeks. These results suggest that enhanced LTP in the SC-CA1 synapses of MSG-induced obese mice involves presynaptic rather than postsynaptic mechanisms., Competing Interests: Declaration of Competing Interest The authors declare no conflicts of interest., (Copyright © 2020 Elsevier B.V. and Japan Neuroscience Society. All rights reserved.)

Background: This study evaluated the relationship between preoperative and postoperative knee kinematics, moreover, investigated tibial rotational position and the extent of tibial internal rotation from knee extension to flexion as factors to obtain significant knee flexion after total knee arthroplasty (TKA)., Methods: Fifty-four patients (60 knees total; 15 males, 16 knees; 39 females, 44 knees) who underwent posterior-stabilized TKA using a navigation system were included. Intraoperative knee kinematics involving tibial rotational position relative to the femur and the extent of tibial internal rotation were examined at two time points: 1) after landmarks registration (pre-TKA) and 2) after skin closure (post-TKA). The relationship between the knee flexion angle at one year postoperatively and intraoperative tibial rotational position, or the extent of tibial rotation among several knee flexion angles calculated with a navigation system were investigated., Results: The postoperative knee flexion angle was positively associated with the preoperative flexion angle and intraoperative knee kinematics at post-TKA involving tibial external position relative to the femur at knee extension and the extent of tibial internal rotation from extension to 90° of flexion or to maximum flexion. There was a positive relationship between the extent of tibial internal rotation at pre-TKA and that at post-TKA., Conclusions: The intraoperative kinematics of the extent of tibial internal rotation at post-TKA was influenced by that at pre-TKA. The greater external position of the tibia relative to the femur at knee extension and the greater extent of tibial internal rotation at post-TKA might lead to good knee flexion angle., Competing Interests: Declaration of competing interest Mitsuru Hanada, Kensuke Hotta, Hiroki Furuhashi, Hiroshi Koyama, and Yukihiro Matsuyama declare that they have no conflict of interest., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Cell movement is crucial for morphogenesis in multicellular organisms. Growing embryos or tissues often expand isotropically, i.e., uniformly, in all dimensions. On the surfaces of these expanding environments, which we call "fields," cells are subjected to frictional forces and move passively in response. However, the potential roles of isotropically expanding fields in morphogenetic events have not been investigated well. Our previous mathematical simulations showed that a tissue was elongated on an isotropically expanding field (Imuta et al., 2014). However, the underlying mechanism remains unclarified, and how cells behave during tissue elongation was not investigated. In this study, we mathematically analyzed the effect of isotropically expanding fields using a vertex model, a standard type of multi-cellular model. We found that cells located on fields were elongated along a similar direction each other and exhibited a columnar configuration with nearly single-cell width. Simultaneously, it was confirmed that the cell clusters were also elongated, even though field expansion was absolutely isotropic. We then investigated the mechanism underlying these counterintuitive phenomena. In particular, we asked whether the dynamics of elongation was predominantly determined by the properties of the field, the cell cluster, or both. Theoretical analyses involving simplification of the model revealed that cell clusters have an intrinsic ability to asymmetrically deform, leading to their elongation. Importantly, this ability is effective only under the non-equilibrium conditions provided by field expansion. This may explain the elongation of the notochord, located on the surface of the growing mouse embryo. We established the mechanism underlying tissue elongation induced by isotropically expanding external environments, and its involvement in collective cell alignment with cell elongation, providing key insight into morphogenesis involving multiple adjacent tissues., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Introduction: Isoproterenol is a non-selective β receptor agonist, which is a drug approved for bradycardia and bronchial asthma in many countries. Recently, isoproterenol has been reported to have the potential as a drug for the treatment of Alzheimer's disease by inhibiting the aggregation of tau protein. Isoproterenol is a highly potent drug causing increases in heart rates even when its plasma concentration is very low. Thus, it is critical to know if potentially effective therapeutic levels of isoproterenol can be achieved, maintaining safe plasma levels without any untoward pharmacological effects. The purpose of the study is to investigate the brain pharmacokinetics and biodistribution of 11 C-labeled isoproterenol in rodents., Methods: We performed positron emission tomography (PET) brain imaging and biodistribution studies of [ 11 C]isoproterenol. 120-min scans with arterial blood sampling were performed in rats. Additionally, plasma and brain homogenates were analyzed with radio-HPLC to characterize its metabolite profiles. As a measure of [ 11 C]isoproterenol brain uptake, total distribution volumes were determined by a pharmacokinetic compartment model. Biodistribution of [ 11 C]isoproterenol was investigated in mice at six-time points from 1-min to 90-min after injection., Results: We found a modest brain uptake of [ 11 C]isoproterenol. Its brain pharmacokinetics showed that the concentration of isoproterenol in the brain at equilibrium was about two-fold higher than in the plasma (total distribution volumes 2.0 ± 0.2 cm 3 /mL). Only unmetabolized isoproterenol was detected in the brain at 30 min after injection, although isoproterenol was rapidly metabolized in plasma. The biodistribution study showed that isoproterenol and its metabolite are excreted mainly via the urinary system. CONCLUSIONS, ADVANCES IN KNOWLEDGE AND IMPLICATIONS FOR PATIENT CARE: In this study, we have shown that rat brain concentrations of isoproterenol are only two-fold of that in plasma at equilibrium. If the brain pharmacokinetics are similar in the human brain, it may be difficult to achieve potentially therapeutic levels of this drug safely in humans. Further studies appear warranted to investigate the brain pharmacokinetics in humans with PET using [ 11 C]isoproterenol., (Copyright © 2020 Elsevier Inc. All rights reserved.)

Pemafibrate, a selective peroxisome proliferator-activated receptor (PPAR) α modulator, is a new drug that specifically modulates PPARα conformation and co-activator recruitment, thereby lowers plasma triglycerides with less off-target effects. Classical PPARα ligands such as fenofibrate suppress inflammatory cells including microglia. However, effects of pemafibrate on microglia have never been addressed. Here we show that pemafibrate, like other PPARα ligands, potently suppressed NF-κB phosphorylation and cytokine expression in microglial cells. PPARα knockdown significantly amplified LPS-induced cytokine expression. Pemafibrate-induced suppression of IL-6 expression was reversed by PPARα knockdown. However, suppression by fenofibrate was not reversed by PPARα knockdown but by Sirtuin 1 (SIRT1) knockdown. In conclusion, pemafibrate and fenofibrate similarly suppresses microglial activation but through distinct PPARα and SIRT1-dependet pathways., Competing Interests: Declaration of competing interest The authors declare no competing interests., (Copyright © 2020 Elsevier Inc. All rights reserved.)

Objective: Exercise training has multiple beneficial effects in patients with arteriosclerotic diseases; however, the exact underlying mechanisms of the effects are not completely understood. This study aimed to evaluate the effectiveness of a supervised exercise program in improving gait parameters, including the variability and walking performance of lower limb movements, in patients with peripheral artery disease (PAD) and intermittent claudication (IC)., Methods: Sixteen patients with a history of PAD and IC were recruited for this study, and they completed a 3-month supervised bicycle exercise program. The ankle-brachial index and responses to quality of life (QOL) questionnaires were evaluated. Near-infrared spectroscopy was also performed to determine the hemoglobin oxygen saturation in the calf. Patients' kinematics and dynamics, including joint range of motion and muscle tension, were evaluated using an optical motion capture system. Computed tomography images of each muscle were assessed by manual outlining. Data were collected before and after the supervised bicycle exercise program, and differences were analyzed., Results: Significant differences were not found in step length, ankle-brachial index, and hemoglobin oxygen saturation before and after the supervised bicycle exercise program; however, IC distance (P = .034), maximum walking distance (P = .006), and all QOL questionnaire scores (P < .001) showed significant improvement. Hip range of motion (P = .035), maximum hip joint torque (right, P = .031; left, P = .044), maximum tension of the gluteus maximus muscle (right, P = .044; left, P = .042), and maximum hip joint work (right, P = .048; left, P = .043) also significantly decreased bilaterally. Computed tomography images showed a significant increase in the cross-sectional area of the abdominal, trunk, and thigh muscles but not in that of the lower leg muscles after the supervised exercise program intervention., Conclusions: In this study, bicycle exercise training improved the QOL and walking distance and decreased hip movement. The results showed that bicycling might be as useful as walking in patients with PAD., (Copyright © 2019 The Authors. Published by Elsevier Inc. All rights reserved.)

A case of small bowel obstruction caused by Bolbosoma sp. infection is reported. A 27-year-old woman admitted with abdominal pain was diagnosed as small bowel obstruction. Laparoscopic surgery revealed induration in jejunum at ca. 120 cm distal to the ligament of Treiz, attributed to a band connecting the serosa to the ascending mesocolon. Resected band contained an acanthocephalan accompanying foreign body reaction with abscess formation. The parasite belonged to the genus Bolbosoma, of which identification was made by DNA sequence analysis. This is the eighth case of Bolbosoma infection in humans, and the first one causing an ileus., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Tubular organs and tissues often show various morphological fold patterns in their luminal epithelia. Computational studies have revealed that these patterns could be explained by mechanical deformation of the epithelia. However, experimental validations of this are sparse, and the mechanisms linking genetic and cellular functions to fold mechanics are poorly understood. In the oviduct of the female reproductive tract, the epithelium forms multiple well-aligned straight folds. Disruption of Celsr1, a planar cell polarity-related gene, causes ectopically-branched folds in mice. Here we discuss the pattern formation of the folds with respect to the growth and mechanics of the epithelium, and the cellular and genetic functions, and compare these with other tubular organs such as the gut., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Purpose: To directly compare fat suppression efficacy of Dixon, spectral attenuated inversion recovery (SPAIR) and short tau inversion recovery (STIR) techniques for a 3T MR system., Materials and Methods: Nineteen consecutive patients (11 men, 8 women; mean age 67.9years) underwent chest MR imaging. Contrast-noise ratio (CNR) between muscle and fat (CNR Fat ) was calculated by ROI measurement. Then, two radiologists used a five-point scale for visual assessment of image quality, fat suppression capability and lesion detection. Next, the quantitative calculations obtained with all three techniques were compared by means of Tukey's honest significant difference (HSD) test. Inter-observer agreements were assessed by kappa statistics and χ 2 test. Fisher's least significant difference (LSD) test was used for comparison among all scores., Results: CNR Fat of SPAIR were significantly higher than those of Dixon and STIR (p<0.001). Inter-observer agreements were assessed as significantly moderate, substantial or almost perfect (0.51<κ<0.89, p<0.0001). Total image quality and fat suppression capability of Dixon and STIR were significantly higher than those of SPAIR (p<0.001)., Conclusion: Dixon technique has better fat suppression capability on T2-weighted turbo SE imaging than SPAIR technique on chest MR imaging examined with a 3T MR system, although it can't substitute STIR technique on fat suppression and lesion visualization., (Copyright © 2017 Elsevier Inc. All rights reserved.)

Background and Aims: Improvement in sleep quality is considered to be a viable target for prevention and treatment of cardiovascular diseases. To gain insight into its underlying mechanisms, we evaluated the significance of objectively measured sleep quality in patients with regard to progression of arterial stiffness over a 3-year follow-up period., Methods: This prospective cohort study included 306 serial patients registered in the Hyogo Sleep Cardio-Autonomic Atherosclerosis (HSCAA) study. In addition to classical cardiovascular risk factors (body mass index, current smoking, past history of cardiovascular disease, dyslipidemia, diabetes mellitus), the participants were examined for ambulatory blood pressure (BP), apnea-hypopnea index (AHI), standard deviation of the NN (RR) interval (SDNN) for heart rate variability (HRV), and objective sleep quality using actigraphy findings. Brachial-ankle pulse wave velocity (baPWV) was measured at both baseline and follow-up (36.6 ± 6.8 months) as a parameter of arterial stiffness., Results: Increases in PWV (%) were greater (p = 0.03) in the low sleep quality (LSQ) group (5.75 ± 1.15%) as compared to the normal sleep quality group (2.69 ± 0.85%). Patients with the greatest increase (≥20%) from baseline exhibited a significantly (p < 0.05) larger percentage of LSQ (75% vs. 49.6%) as compared to those without PWV progression (<0%), with the association still significant (odds ratio 3.62, 95% confidence interval 1.04-12.55, p = 0.04) even after adjustment for other clinical risk factors. For all subjects, univariate logistic regression analyses showed that diabetes and LSQ were significantly associated with the greatest increase of PWV. Comparisons of characteristics among specific subgroups showed more prominent associations of LSQ with the greatest increase of PWV in patients with greater age, dyslipidemia, and higher AHI., Conclusions: LSQ was associated with progression of arterial stiffness over a 3-year period, independent of cardiovascular risk factors such as BP, AHI, and HRV., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Medaka (Oryzias latipes) is a temperate eurythermal fish that is able to survive over a wide range of water temperatures ranging from near zero to over 30°C throughout the year; it maintains its normal physiological and biochemical processes through temperature acclimation. To determine the mechanisms involved in temperature acclimation of fish, the fast skeletal muscle tissues of medaka underwent global gene expression analysis using next-generation sequencing. Ten individuals were placed into two aquariums at 24°C. While the water temperature of one aquarium was decreased to 10°C, that of the other aquarium was increased to 30°C; these temperatures were subsequently maintained for 5weeks. RNA sequencing (RNA-Seq) analyses revealed that 11 genes involved in energy metabolism and muscle atrophy were significantly highly expressed in the 10°C-acclimated fish. Meanwhile, significantly higher expression levels were observed for 20 genes encoding myofibrillar proteins and heat shock proteins in the 30°C-acclimated fish. Moreover, 1103 genes had at least fourfold differential expression between the acclimation groups. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses provided important information: although the expression of genes related to metabolic processes were activated, muscle atrophy occurred in the 10°C-acclimated fish, and muscle cells divided actively in the 30°C-acclimated fish and avoided thermal stress by expressing heat shock proteins. Therefore, RNA-Seq analyses with the available genome database will be useful for better understanding the molecular mechanisms involved in the temperature acclimation of fish., (Copyright © 2017 Elsevier Inc. All rights reserved.)

Background: In total hip arthroplasty (THA) surgery, hip muscle preservation is important in strengthening the stability of the hip and improving the activities of the patient. However, whether the type of femoral stem affects the recovery of the hip muscles remains unknown. The aim of this study was to compare the postoperative hip muscle recovery among femoral stem varieties after THA., Methods: The computed tomography (CT) images of 44 patients (44 hips) who underwent THA using an anterolateral approach were reviewed. Twenty-two patients received a fit-and-fill (FF) stem and 22 received the tapered-wedge (TW) stem. The volumes of the gluteus maximus (GMA), gluteus medius (GME), and obturator internus (OI) were measured on three-dimensional models reconstructed using preoperative and 6-month postoperative CT images. Relationships between muscle volume changes and factors including the femoral stem length were evaluated., Results: The GMA and GME volumes increased postoperatively by 8.2% and 8.3%, respectively, in the FF stem group and 7% and 6%, respectively, in the TW stem group, with no group differences. In contrast, the OI volume decreased postoperatively by 17.8% in the FF group and was preserved in TW group (p < 0.001). Moreover, OI volume was decreased in 19 patients (86%) in the FF group and in 11 patients (50%) in the TW group (p = 0.01). The normalized stem length was significantly associated with the postoperative change in OI volume (r = -0.45, p = 0.002)., Conclusions: The TW stem showed a significant advantage over the FF stem in terms of OI preservation. Surgeons should pay close attention during surgery to avoid OI injury when using different femoral stem types. We suggest that a short and reduced lateral shoulder femoral stem is a better choice for the preservation of external rotation muscles., (Copyright © 2017 The Japanese Orthopaedic Association. Published by Elsevier B.V. All rights reserved.)

Background: Excessive mucin secretion in the airway is an important feature of airway inflammatory diseases. MUC5AC expression is regulated by a variety of stimuli such as cytokines. Little is known about the role of interferon (IFN)-γ in MUC5AC expression in human bronchial epithelial cells., Methods: Human pulmonary mucoepidermoid carcinoma cell line (NCI-H292) and normal human bronchial epithelial (NHBE) cells were used to assess the effects of IFN-γ on MUC5AC transcription., Results: Transforming growth factor (TGF)-α and double-stranded RNA (polyI:C)-induced MUC5AC mRNA and protein expression was repressed by IFN-γ in a concentration-dependent manner. IFN-γ showed limited effects on TGF-α and polyI:C-induced activation of epidermal growth factor receptor (EGFR) and extracellular signal-regulated kinase (ERK). A chromatin immunoprecipitation assay indicated that Sp1 bound to its cognate sequence located on the MUC5AC promoter. The Sp1 inhibitor mithramycin A inhibited MUC5AC mRNA expression, implying a critical role for Sp1 in MUC5AC induction. Importantly, IFN-γ impeded Sp1 binding to the MUC5AC promoter., Conclusions: These results suggest that IFN-γ represses MUC5AC expression, disturbing binding of Sp1 to its target sequences., (Copyright © 2016 Japanese Society of Allergology. Production and hosting by Elsevier B.V. All rights reserved.)

Introduction: Determine the usefulness of dyssynchrony indices derived from two-dimensional speckle tracking echocardiography for the detection of mechanical dyssynchrony in a canine model of left bundle branch block., Animals: Ten healthy beagles., Materials and Methods: Segmental, time-radial strain curves were obtained using two-dimensional speckle tracking echocardiography. The maximum difference and standard deviation of the time to peak radial strain for six predefined segments (MaxD-TpSR and 6SD-TpSR) were calculated, together with the left ventricular dyssynchrony by radial strain (DysSR), before and after ablation of the left bundle branch block. Receiver operating characteristic curve analysis was performed using dogs after ablation as positive controls., Results: After ablation, all dogs showed multiple peaks in at least one segment on the time-radial strain curve, while all dyssynchrony indices increased significantly (MaxD-TpSR from 16.25 ± 16.04 [mean ± standard deviation] to 44.4 ± 26.18 ms, 6SD-TpSR from 7.59 ± 7.40 to 19.62 ± 11.91 ms, and DysSR from 4.20 ± 2.12 to 10.87± 2.92%, p<0.05). In receiver operating characteristic curve analysis, the areas under the curve for MaxD-TpSR, 6SD-TpSR, and DysSR were 0.825, 0.800, and 0.980, respectively., Conclusions: Left ventricular dyssynchrony by radial strain can detect mechanical dyssynchrony with higher sensitivity and specificity than dyssynchrony indices, based on the time to peak radial strain., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Objectives: The objective of our study was to quantitatively measure systolic torsional deformations in cats with hypertrophic cardiomyopathy (HCM) and in controls., Animals: Twenty-six client-owned cats with HCM and 14 healthy cats. HCM cats were categorized based on their symptoms (asymptomatic and symptomatic) and with or without left ventricular outflow tract obstruction (obstructive and non-obstructive)., Methods: The cats were examined for myocardial deformations using two-dimensional speckle-tracking echocardiography and were evaluated for peak systolic rotation and the rotation rate at each basal and apical view. Cats were also evaluated for the peak systolic torsion and torsion rate., Results: The peak systolic apical rotation and torsion were higher in asymptomatic and symptomatic cats with HCM than in control cats. Also, the peak systolic apical rotation, apical rotation rate, torsion, and torsion rate were higher in cats with obstructive HCM than in control cats., Conclusions: Myocardial torsional deformations assessed by two-dimensional speckle-tracking echocardiography may be useful for evaluating compensatory myocardial function of HCM., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Despite extensive studies on clinical indices for the selection of patient candidates for cardiac resynchronization therapy (CRT), approximately 30% of selected patients do not respond to this therapy. Herein, we examined whether CRT simulations based on individualized realistic three-dimensional heart models can predict the therapeutic effect of CRT in a canine model of heart failure with left bundle branch block. In four canine models of failing heart with dyssynchrony, individualized three-dimensional heart models reproducing the electromechanical activity of each animal were created based on the computer tomographic images. CRT simulations were performed for 25 patterns of three ventricular pacing lead positions. Lead positions producing the best and the worst therapeutic effects were selected in each model. The validity of predictions was tested in acute experiments in which hearts were paced from the sites identified by simulations. We found significant correlations between the experimentally observed improvement in ejection fraction (EF) and the predicted improvements in ejection fraction (P<0.01) or the maximum value of the derivative of left ventricular pressure (P<0.01). The optimal lead positions produced better outcomes compared with the worst positioning in all dogs studied, although there were significant variations in responses. Variations in ventricular wall thickness among the dogs may have contributed to these responses. Thus CRT simulations using the individualized three-dimensional heart models can predict acute hemodynamic improvement, and help determine the optimal positions of the pacing lead., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Objective: We previously observed that silencing of eukaryotic translation initiation factor 3 subunit e (eIF3e), a hypoxia-independent downregulator of hypoxia-inducible factor 2α (HIF-2α), led to neoangiogenesis by promoting HIF-2α activity under normoxic conditions. In the current study, we investigated whether temporary silencing of eIF3e in muscles affects blood flow recovery in a mouse ischemic limb model., Methods: eIF3e silencing was performed using small interfering RNA (siRNA), and changes in gene transcription and protein expression were analyzed in vitro using murine primary skeletal muscle myoblast and human primary skeletal muscle myoblast cell cultures. In unilateral femoral artery ligation experiments, eIF3e siRNA-expressing plasmids were injected into the muscles of BALB/c mice near the ligation sites, and tissue damage and loss of limb function were scored for 28 days while serial measurements of limb perfusions were performed with laser Doppler perfusion imaging., Results: Silencing of eIF3e in murine primary skeletal muscle myoblasts led to stabilization of HIF-2α and upregulation of angiogenic transcripts, including basic fibroblast growth factor and platelet-derived growth factor B (P < .05), and the supernatant of eIF3e-silenced human primary skeletal muscle myoblasts triggered the tube formation of human umbilical vein endothelial cells. The in vivo mouse model of hindlimb ischemia revealed that single intramuscular injections of eIF3e siRNA-expressing plasmids significantly enhanced perfusion of ischemia-damaged limbs (P < .05) at days 7 and 14 and functional recovery at days 7, 14, and 21 (P < .05)., Conclusions: eIF3e is an angiogenesis suppressor and may be a therapeutic target for promoting angiogenesis after ischemic injuries., (Copyright © 2016 Society for Vascular Surgery. Published by Elsevier Inc. All rights reserved.)

Background: Induction of collateral development to improve tissue perfusion is a promising approach for the treatment of arterial occlusive diseases. Several growth factors and cells have been reported to increase collateral circulation; however, the appropriate site for the delivery of these factors and cells is unclear. In this study, we identified the delivery site for growth factor in a rabbit model of limb ischemia and evaluated whether specific delivery of basic fibroblast growth factor (bFGF) to this site enhanced collateral augmentation., Methods: The left femoral artery of Japanese white rabbits was excised to induce limb ischemia. Twenty-eight days thereafter, angiograms were obtained to identify the typical pattern of collateral development in this model. Subsequently, bFGF (100 μg) was selectively injected into the left coccygeofemoral muscle (coccygeo group) or adductor muscle (adductor group), major thigh muscles in proximity. Collateral development was evaluated at 28 days after injection, and its mechanism was assessed by immunologic and morphometric analyses of muscle samples., Results: Angiographic evaluation of this model revealed that after femoral artery excision, collateral vessels generally developed in the left coccygeofemoral muscle, whereas few collateral vessels were detected in the left adductor muscle. At 28 days after injection, calf blood pressure ratio, defined as left pressure to right pressure, was significantly higher in the coccygeo group than in the adductor group (0.85 ± 0.05 vs 0.69 ± 0.05, respectively; P < .01). Similar results were observed in blood flow through the internal iliac artery (resting: 24.6 ± 6.1 vs 17.4 ± 8.0 mL/min, P < .01; maximum: 47.4 ± 12.3 vs 33.2 ± 10.7 mL/min, P < .01) and in the angiographic score (0.67 ± 0.13 vs 0.39 ± 0.11; P < .01). Immunologic analyses of the coccygeofemoral muscle at day 3 showed marked expressions of Ki-67, monocyte chemotactic protein 1, and FGF receptor 1 in the coccygeo group compared with the adductor group. Morphometric analyses of the same muscle at day 14 also revealed that collateral vessel density and wall thickness were significantly increased in the coccygeal group compared with the adductor group., Conclusions: These findings demonstrated that selective bFGF delivery to the coccygeofemoral muscle markedly improved collateral development and limb perfusion compared with delivery to the adductor muscle, suggesting that site selection is important in increasing therapeutic efficacy., (Copyright © 2015 Society for Vascular Surgery. Published by Elsevier Inc. All rights reserved.)

Methicillin-resistant Staphylococcus aureus (MRSA) has now been recognized as a common pathogen in the community. Sequence type (ST) 398 MRSA is generally considered as an emerging zoonotic agent spreading among livestock and personnel who have direct contact with animals, mainly in Europe. A 37-year-old Chinese woman receiving steroid therapy for systemic lupus erythematosus with general fatigue and myalgia was brought to the emergency department in critical condition. Her condition deteriorated despite aggressive management and she died on day 7. Her blood culture revealed ST398 MRSA-SCCmec V with Panton-Valentine Leukocidin (PVL) gene. This is the first case report of a fatal infection caused by this lineage. According to the results of molecular analyses, the isolate from this particular patient's blood was genetically close to a lineage detected in China, and is less likely to be related to an animal-associated lineage., (Copyright © 2015 Japanese Society of Chemotherapy and The Japanese Association for Infectious Diseases. Published by Elsevier Ltd. All rights reserved.)

Hypothermia can occur during fasting when thermoregulatory mechanisms, involving fatty acid (FA) utilization, are disturbed. CD36/FA translocase is a membrane protein which facilitates membrane transport of long-chain FA in the FA consuming heart, skeletal muscle (SkM) and adipose tissues. It also accelerates uptake of triglyceride-rich lipoprotein by brown adipose tissue (BAT) in a cold environment. In mice deficient for CD36 (CD36(-/-) mice), FA uptake is markedly reduced with a compensatory increase in glucose uptake in the heart and SkM, resulting in lower levels of blood glucose especially during fasting. However, the role of CD36 in thermogenic activity during fasting remains to be determined. In fasted CD36(-/-) mice, body temperature drastically decreased shortly after cold exposure. The hypothermia was accompanied by a marked reduction in blood glucose and in stores of triacylglycerols in BAT and of glycogen in glycolytic SkM. Biodistribution analysis using the FA analogue (125)I-BMIPP and the glucose analogue (18)F-FDG revealed that uptake of FA and glucose was severely impaired in BAT and glycolytic SkM in cold-exposed CD36(-/-) mice. Further, induction of the genes of thermogenesis in BAT was blunted in fasted CD36(-/-) mice after cold exposure. These findings strongly suggest that CD36(-/-) mice exhibit pronounced hypothermia after fasting due to depletion of energy storage in BAT and glycolytic SkM and to reduced supply of energy substrates to these tissues. Our study underscores the importance of CD36 for nutrient homeostasis to survive potentially life-threatening challenges, such as cold and starvation., (Copyright © 2015 Elsevier Inc. All rights reserved.)

Objectives: Behavioral and psychosocial factors have been gaining increased attention in regard to cardiovascular diseases. We evaluated sleep conditions, cardiac autonomic function, and carotid atherosclerosis in subjects who participated in the Hyogo Sleep Cardio-Autonomic Atherosclerosis (HSCAA) Study., Methods: This cross-sectional study included 330 serial patients registered in the HSCAA study who were free from past cardiovascular diseases, and prescribing α- or β-blockers. In addition to clinical background and classical cardiovascular risk factors, sleep efficiency, apnea hypopnea index (AHI), awake physical activity, heart rate variability (HRV), carotid intima-media thickness (IMT), presence of plaque and plaque score were determined., Results: Sleep efficiency (r = -0.183) and all HRV parameters (SDNN: r = -0.202; rMSSD: r = -0.234; pNN50: r = -0.277) were significantly (p < 0.01) and negatively associated with IMT, while AHI (r = 0.220, p < 0.001) was positively associated with IMT. Similarly, sleep efficiency (r = -0.129), HRV parameters (SDNN: r = -0.170; rMSSD: r = -0.217; pNN50: r = -0.260) and AHI (r = 0.184) were also significantly (p < 0.05) associated with plaque scores. Multivariate logistic regression analyses showed that rMSSD, but not sleep efficiency or AHI, was significantly associated with carotid plaque (OR 0.74, 95% CI 0.56-0.98, p = 0.037), independent of classical risk factors. The association of rMSSD with carotid plaque remained significant even after adjustment for sleep efficiency or AHI. A comparison of risk factors in specific subgroups showed that the association of lower HRV with carotid plaque was more prominent in patients with cardiovascular risk factors including male gender, hypertension, dyslipidemia and diabetes mellitus., Conclusion: Cardiac autonomic nervous dysfunction was independently associated with carotid atherosclerosis, independent of sleep condition. Moreover, that association was more prominent in specific subgroups with cardiovascular risk factors., (Copyright © 2014 Elsevier Ireland Ltd. All rights reserved.)

The clinical success of cell-based therapeutic angiogenesis has been limited in diabetic patients with critical limb ischemia. We previously reported that an injectable cell scaffold (ICS), which is a nano-scaled hydroxyapatite (HAp)-coated polymer microsphere, enhances therapeutic angiogenesis. Subsequently, we developed a modified ICS for clinical use, measuring 50 μm in diameter using poly(l-lactide-co-ε-caprolactone) as a biodegradable polymer, which achieved appropriately accelerated absorption in vivo. The aim of the present study was to evaluate the effectiveness of this practical ICS in diabetic hindlimb ischemia. Bone-marrow mononuclear cells (BMNCs) were intramuscularly injected, without or with a practical ICS, into the ischemic hindlimbs of mice (BMNCs or ICS+BMNCs group, respectively). Kaplan-Meier analysis demonstrated that the beneficial effects of BMNC transplantation for limb salvage after ischemic surgery were almost entirely abrogated in streptozotocin-induced diabetic mice. In contrast, injection of ICS+BMNCs revealed significant limb salvage in diabetic mice to a similar extent as in non-diabetic mice. The number of apoptotic transplanted BMNCs was 1.8-fold higher in diabetic mice 10 days after transplantation compared to non-diabetic mice, while that in the ICS+BMNCs group was markedly lower (8.3% of that in the BMNCs group) even in diabetic mice. The proangiogenic factors VEGF and FGF2, also known as antiapoptotic factors, mostly co-localized with transplanted GFP-positive BMNCs that were closely aggregated around the ICS in ischemic tissue. In conclusion, the practical ICS significantly augmented cell-based therapeutic angiogenesis even in diabetic animals, through local accumulation of proangiogenic factors and antiapoptotic effects in transplanted cells., (Copyright © 2014 Elsevier Inc. All rights reserved.)

Ameloblastoma is an odontogenic benign tumor that occurs in the jawbone, which invades bone and reoccurs locally. This tumor is treated by wide surgical excision and causes various problems, including changes in facial countenance and mastication disorders. Ameloblastomas have abundant tumor stroma, including fibroblasts and immune cells. Although cell-to-cell interactions are considered to be involved in the pathogenesis of many diseases, intercellular communications in ameloblastoma have not been fully investigated. In this study, we examined interactions between tumor cells and stromal fibroblasts via soluble factors in ameloblastoma. We used a human ameloblastoma cell line (AM-3 ameloblastoma cells), human fibroblasts (HFF-2 fibroblasts), and primary-cultured fibroblasts from human ameloblastoma tissues, and analyzed the effect of ameloblastoma-associated cell-to-cell communications on gene expression, cytokine secretion, cellular motility and proliferation. AM-3 ameloblastoma cells secreted higher levels of interleukin (IL)-1α than HFF-2 fibroblasts. Treatment with conditioned medium from AM-3 ameloblastoma cells upregulated gene expression and secretion of IL-6 and IL-8 of HFF-2 fibroblasts and primary-cultured fibroblast cells from ameloblastoma tissues. The AM3-stimulated production of IL-6 and IL-8 in fibroblasts was neutralized by pretreatment of AM-3 cells with anti-IL-1α antibody and IL-1 receptor antagonist. Reciprocally, cellular motility of AM-3 ameloblastoma cells was stimulated by HFF-2 fibroblasts in IL-6 and IL-8 dependent manner. In conclusion, ameloblastoma cells and stromal fibroblasts behave interactively via these cytokines to create a microenvironment that leads to the extension of ameloblastomas., (Copyright © 2014 Elsevier Inc. All rights reserved.)

Objective: The receptor for advanced glycation end-products (RAGE) is involved in vascular complications in diabetic patients. Pioglitazone, in contrast to glimepiride, has been shown to be protective against atherosclerotic disorders. In this study, we directly compared the effects of those drugs on RAGE system., Methods: Sixty-three type 2 diabetic patients (age 20-80 years, hemoglobin A1c 6.4-10.3%) being treated with sulfonylurea (glimepiride 0.5-2.0 mg/day, glyclazide 20-80 mg/day, glibenclamide 1.25-5.0 mg/day), or with nateglinide or metiglynide were randomly assigned to receive either pioglitazone (n = 31) or glimepiride (n = 32). Levels in plasma of soluble RAGE (sRAGE) and endogenous secretory RAGE (esRAGE), and RAGE expression in peripheral mononuclear cells were determined at 0, 12, and 24 weeks., Results: Twenty-seven patients in the pioglitazone group (15-30 mg) and 30 in the glimepiride group (0.5-4 mg) completed the 24-week trial. Increases in plasma esRAGE were significantly greater in the pioglitazone group (12 weeks: 55 ± 15 pg/mL, p = 0.018; 24 weeks: 90 ± 14 pg/mL, p = 0.003) as compared to the glimepiride group (12 weeks: 12 ± 9 pg/mL; 24 weeks: 29 ± 14 pg/mL). Increases in plasma sRAGE were also significantly (p = 0.037) higher in the pioglitazone group at 24 weeks (170 ± 166 vs.74 ± 171 pg/mL). Furthermore, RAGE expression in mononuclear cells was significantly (p = 0.008) decreased to a greater degree in the pioglitazone group at 24 weeks (-7.39 ± 5.18 vs. -3.39 ± 5.72 MFI). Changes in HbA1c, IRI, and insulin resistance index (HOMA) at 24 weeks were not significantly different between the groups., Conclusion: Pioglitazone suppresses RAGE expression and increases circulating sRAGE/esRAGE, and those activities are not necessarily dependent on plasma glucose or insulin resistance levels., Clinical Trial No: UMIN000002055., (Copyright © 2014 Elsevier Ireland Ltd. All rights reserved.)

Mammalian embryos develop in coordination with extraembryonic tissues, which support embryonic development by implanting embryos into the uterus, supplying nutrition, providing a confined niche, and also providing patterning signals to embryos. Here, we show that in mouse embryos, the expansion of the amniotic cavity (AC), which is formed between embryonic and extraembryonic tissues, provides the mechanical forces required for a type of morphogenetic movement of the notochord known as convergent extension (CE) in which the cells converge to the midline and the tissue elongates along the antero-posterior (AP) axis. The notochord is stretched along the AP axis, and the expansion of the AC is required for CE. Both mathematical modeling and physical simulation showed that a rectangular morphology of the early notochord caused the application of anisotropic force along the AP axis to the notochord through the isotropic expansion of the AC. AC expansion acts upstream of planar cell polarity (PCP) signaling, which regulates CE movement. Our results highlight the importance of extraembryonic tissues as a source of the forces that control the morphogenesis of embryos., (Copyright © 2014 Elsevier Ireland Ltd. All rights reserved.)

Background: The forced oscillation technique (FOT) is useful for studying pulmonary function in children, as well as in school children with asthma. However, the standard values for respiratory resistance (Rrs) in Asian school children remain unknown. We evaluated the standard Rrs using a type of FOT, impulse oscillometry (IOS), in healthy Japanese children at elementary and junior high schools., Methods: A total of 795 children (age range: 6-15 years; mean age ± SD: 11.1 ± 2.4 years; 404 boys, 391 girls) at elementary and junior high schools participated in the study. Of the 795 children, we evaluated the Rrs of 537 children aged 6-15 years (mean ± SD: 10.8 ± 2.4 years) using IOS., Results: Regression analyses with three IOS parameters, Rrs at 5Hz (R5), Rrs at 20Hz (R20), and Rrs difference between 5Hz and 20Hz (R5-R20), for age, height, weight, and degree of obesity as independent variables demonstrated the strongest correlation between each parameter and children's height. All parameters decreased with increasing height. Using the lambda-mu-sigma (LMS) method, we created standard curves for the Rrs values based on height., Conclusions: Our standard curves could be useful for diagnosis and control evaluation of childhood asthma.

Objectives: The objective of this study was to evaluate the effect of age on myocardial function assessed by two-dimensional speckle-tracking echocardiography (2D-STE) in healthy dogs., Animals: Thirty-two healthy Beagles were used., Methods: Myocardial function was assessed in each dog by using 2D-STE, and the results were compared between young and old dogs., Results: The myocardial deformations in systole, besides the apical rotation rate, were not significantly different between young and old dogs. In contrast, the early diastolic circumferential strain rate, basal rotation rate, and torsion rate were significantly lower in old dogs than in young dogs (P = 0.03, P = 0.033, and P = 0.015, respectively). Late diastolic longitudinal and radial strain rates were significantly higher in old dogs than in young dogs (P = 0.002 and P = 0.018, respectively)., Conclusions: Young and old dogs showed similar systolic myocardial deformations, but significant differences in the values of some diastolic deformation variables were found between young and old dogs, highlighting the need for using age-matched control subjects in studies of diastolic function., (Copyright © 2013 Elsevier B.V. All rights reserved.)

Objectives: To determine the quantitative diagnostic capability of magnetic resonance imaging (MRI), compared to dual-energy X-ray absorptiometry (DEXA), for osteoporosis., Materials and Methods: Eight male volunteers and eight patients underwent both DEXA and MRI. Results were obtained from each subject's L2 to L4, for a total of 48 lumbar vertebrae. Based on their bone mineral density (BMD) acquired from DEXA, the vertebrae were classified as follows: normal (n=28), osteopenic (n=0), and osteoporotic (n=20). All MR examinations were performed on a 1.5-T scanner to obtain T1-weighted imaging (T1WI), fat-suppression T2-weighted imaging (FS-T2WI), and diffusion-weighted imaging (DWI). These quantitative MR parameters were determined: T1WI and FS-T2WI signal-to-noise ratios and DWI apparent diffusion coefficient values. To determine the diagnostic capability of MRI as an osteoporosis indicator, MR parameters were assessed statistically., Results: All MR parameters significantly correlated with BMD (T1WI: r=-0.64, FS-T2WI: r=-0.36, DWI: r=-0.29), with significant differences among normal and osteoporotic vertebrae (P<.05). By receiver operating characteristic analysis, the area under the curve of T1WI was significantly higher than others (P<.05). When adapted as feasible threshold values, sensitivity, specificity, and accuracy of T1WI were 95.0% (19/20), 92.9% (26/28), and 93.8% (45/48), respectively., Conclusion: Routine MRI, in particular T1WI, had a potential for the assessment of osteoporosis., (Copyright © 2013 Elsevier Inc. All rights reserved.)

Objective: The objective of this study was to evaluate the influence of heart rate (HR) on myocardial function assessed by two-dimensional speckle-tracking echocardiography (2D-STE) in healthy dogs., Animals: Thirteen healthy beagle dogs., Methods: Animals were anesthetized and HR was controlled with right atrial pacing. Myocardial function of each dog was assessed using 2D-STE at pacing rates of 120, 140, 160, and 180 bpm., Results: All strain and strain rate variables in the longitudinal, circumferential, and radial directions were not significantly different between pacing rates. Peak early diastolic torsion rate at 180 bpm was significantly increased compared with that at 120 bpm (P = 0.003)., Conclusion: Torsion rate in early diastole was elevated at 180 bpm, which may reflect improved myocardial relaxation with higher HR. Changes in left ventricular torsion during tachycardia may play an important role in preserving stroke volume in the presence of shortened ejection and filling times., (Copyright © 2013 Elsevier B.V. All rights reserved.)

Objective: The objective of this study was to clinically assess myocardial deformations in dogs with chronic mitral valve insufficiency (CMVI) using two-dimensional speckle-tracking echocardiography (2D-STE)., Animals: 87 dogs with CMVI., Methods: Dogs were placed into 1 of 3 classes, based on the International Small Animal Cardiac Health Council classification. In addition, 20 weight- and age-matched healthy dogs were enrolled as controls. The dogs were examined for myocardial deformations using 2D-STE, and strain and strain rate in the longitudinal, circumferential, and radial directions were evaluated., Results: Class II and III dogs had higher circumferential strain than class I dogs (P = 0.002 and P = 0.001, respectively) and controls (P < 0.001 and P < 0.001, respectively). Class III dogs had higher radial strain than class I dogs (P = 0.001) and controls (P < 0.001). Class III dogs had higher radial strain rate than class I dogs (P = 0.006) and controls (P = 0.001). Other deformations, including longitudinal deformations, were not significantly different between classes of CMVI or between CMVI dogs and controls., Conclusions: In the clinical progression of CMVI in dogs, myocardial deformations, as assessed by 2D-STE, differed according to myocardial contractile direction. Thus, assessments of multidirectional myocardial deformations may be important for better assessment of clinical cardiac function in dogs with CMVI., (Copyright © 2013 Elsevier B.V. All rights reserved.)

Influenza viruses have developed resistance to current drugs, creating a need for new antiviral targets and new drugs to treat influenza virus infections. In this study, computational and experimental screening of an extensive compound library identified THC19, which was able to suppress influenza virus replication. This compound had no cytotoxic effects and did not disrupt cell cycle progression or induce apoptosis in MDCK cells as confirmed by WST-1 assays, flow cytometry analysis, and caspase-3 assays. Time-of-addition experiments showed that THC19 acts at a relatively early stage of the viral lifecycle. Subsequent mini-genome assays revealed that THC19 inhibited viral genome replication and/or transcription, suggesting that it interferes with one or more of the viral components that form the ribonucleoprotein complexes, namely polymerase basic 2 (PB2), polymerase basic 1 (PB1), polymerase acidic (PA), nucleoprotein (NP) and viral RNA. Finally, mini-genome assays where PB2, PB1, PA or NP from A/WSN/33 (H1N1) virus were replaced with those from A/Udorn/307/1972 (H3N2) virus effectively demonstrated that THC19 inhibited viral multiplication in a manner dependent upon the PA subunit. Taken together, these results suggest that influenza virus PA protein is a potential target for, and may aid the development of, novel compounds that inhibit influenza A virus replication., (Copyright © 2012 Institut Pasteur. Published by Elsevier Masson SAS. All rights reserved.)

Using Tribolium castaneum, we quantitatively investigated the induction of nine antimicrobial peptide (AMP) genes by live gram-negative bacteria (Escherichia coli and Enterobacter cloacae), gram-positive bacteria (Micrococcus luteus and Bacillus subtilis) and the budding yeast (Saccharomyces cerevisiae). Then, five representative AMP genes were selected, and the involvement of the Toll and IMD pathways in their induction by E. coli, M. luteus and S. cerevisiae was examined by utilizing RNA interference of either MyD88 or IMD. Results indicated: Robust and acute induction of three genes by the two bacterial species was mediated mainly by the IMD pathway; slow and sustained induction of one gene by the two bacteria was mediated mainly by the Toll pathway; induction of the remaining one gene by the two bacteria was mediated by both pathways; induction of the five genes by the yeast was mediated by the Toll and/or IMD pathways depending on respective genes. These results suggest that more promiscuous activation and usage of the two pathways may occur in T. castaneum than in Drosophila melanogaster. In addition, the IMD pathway was revealed to dominantly contribute to defense against two bacterial species, gram-negative E. cloacae and gram-positive B. subtilis that possesses DAP-type peptidoglycan.

Phenylenediamine derivatives can function as a hydrogen donor and reportedly exert various biological actions including cytoprotective effects against oxidative stress, possibly by acting as an antioxidant. Previous studies showed that feeding of such compounds to mice reduced their body weight, but the precise mechanism remains unknown at present. Here, we found that these compounds inhibited the in vitro differentiation of mouse preadipocytes, 3T3-L1 cells, into adipocytes, suggesting that, at least in part, reduced generation of adipocytes might contribute to the observed weight loss in mice. Next, we performed array analysis and found that the expression of GDF-15/MIC-1, which is a TGFβ superfamily cytokine, and Trib 3, an intracellular downstream effector of the cytokines, was up-regulated by these derivatives. Thus, we identified the compounds as inducers of GDF-15/MIC-1 and suggest that such induction may have led to inhibition of adipocyte differentiation, which could account for the weight-loss effect of these compounds., (Copyright © 2011 Elsevier Inc. All rights reserved.)