Assessing Molecular Localization of Symbiont Microalgae in Coral Branches Through Mass Spectrometry Imaging.

Reef-building corals are a fundamental pillar of coral reef ecosystems in tropical and subtropical shallow environments. Corals harbor symbiotic dinoflagellates belonging to the family Symbiodiniaceae, commonly known as zooxanthellae. Extensive research has been conducted on this symbiotic relationship, yet the fundamental information about the distribution and localization of Symbiodiniaceae cells in corals is still limited. This information is crucial to understanding the mechanism underlying the metabolite exchange between corals and their algal symbionts, as well as the metabolic flow within holobionts. To examine the distribution of Symbiodiniaceae cells within corals, in this study, we used fluorescence imaging and matrix-assisted laser desorption/ionization (MALDI) mass spectrometry imaging (MS-Imaging) on branches of the Acropora tenuis coral. We successfully prepared frozen sections of the coral for molecular imaging without fixing or decalcifying the coral branches. By combining the results of MS-Imaging with that of the fluorescence imaging, we determined that the algal Symbiodiniaceae symbionts were not only localized in the tentacle and surface region of the coral branches but also inhabited the in inner parts. Therefore, the molecular imaging technique used in this study could be valuable to further investigate the molecular dynamics between corals and their symbionts.

Modeling the musculoskeletal system of an insect thorax for flapping flight.

Indirect actuation of the wings via thoracic deformation is a unique mechanism widely observed in flying insect species. The physical properties of the thorax have been intensively studied in terms of their ability to efficiently generate wingbeats. The basic mechanism of indirect wing actuation is generally explained as a lever model on a cross-sectional plane, where the dorsoventral movement of the mesonotum (dorsal exoskeleton of the mesothorax) generated by contractions of indirect muscles actuates the wing. However, the model considers the mesonotum as an ideal flat plane, whereas the mesonotum is hemispherical and becomes locally deformed during flight. Furthermore, the conventional model is two-dimensional; therefore, three-dimensional wing kinematics by indirect muscles have not been studied to date. In this study, we develop structural models of the mesonotum and mesothorax of the hawkmothAgrius convolvuli, reconstructed from serial cross-sectional images. External forces are applied to the models to mimic muscle contraction, and mesonotum deformation and wing trajectories are analyzed using finite element analysis. We find that applying longitudinal strain to the mesonotum to mimic strain by depressor muscle contraction reproduces local deformation comparable to that of the thorax during flight. Furthermore, the phase difference of the forces applied to the depressor and elevator muscles changes the wing trajectory from a figure eight to a circle, which is qualitatively consistent with the tethered flight experiment. These results indicate that the local deformation of the mesonotum due to its morphology and the thoracic deformation via indirect power muscles can modulate three-dimensional wing trajectories.

ViBrism DB: an interactive search and viewer platform for 2D/3D anatomical images of gene expression and co-expression networks.

Understanding anatomical structures and biological functions based on gene expression is critical in a systemic approach to address the complexity of the mammalian brain, where >25 000 genes are expressed in a precise manner. Co-expressed genes are thought to regulate cell type- or region-specific brain functions. Thus, well-designed data acquisition and visualization systems for profiling combinatorial gene expression in relation to anatomical structures are crucial. To this purpose, using our techniques of microtomy-based gene expression measurements and WebGL-based visualization programs, we mapped spatial expression densities of genome-wide transcripts to the 3D coordinates of mouse brains at four post-natal stages, and built a database, ViBrism DB (http://vibrism.neuroinf.jp/). With the DB platform, users can access a total of 172 022 expression maps of transcripts, including coding, non-coding and lncRNAs in the whole context of 3D magnetic resonance (MR) images. Co-expression of transcripts is represented in the image space and in topological network graphs. In situ hybridization images and anatomical area maps are browsable in the same space of 3D expression maps using a new browser-based 2D/3D viewer, BAH viewer. Created images are shareable using URLs, including scene-setting parameters. The DB has multiple links and is expandable by community activity.

A hepatic pDNA delivery system based on an intracellular environment sensitive vitamin E-scaffold lipid-like material with the aid of an anti-inflammatory drug.

Non-viral vectors are considered to be an attractive approach for gene delivery, since an artificial material is less immunogenic and oncogenic compared to a viral vector. We previously reported on the hepatic delivery of plasmid DNA (pDNA) by using lipid-like material (an SS-cleavable and pH-activated lipid-like material: ssPalm) which mounts two hydrophobic scaffolds, proton-accepting motifs (tertiary amines), and a cleavable unit (disulfide bonding). In the present study, we report on an advanced hepatic gene delivery system that uses a new type of ssPalm derivative: ssPalmE-Paz4-C2. The hepatic transgene expression of the intravenously administrated lipid nanoparticle (LNP) that was formed with the ssPalmE-Paz4-C2 (LNPssPalmE-Paz4-C2) was significantly higher than that of conventional LNPs formed with a myristic acid-scaffold ssPalm (LNPssPalmM). However, the LNPssPalmE-Paz4-C2 particle induced a severe innate immune response that involved the production of the pro-inflammatory cytokines (IL-6 and TNFα), intracellular DNA sensor-related cytokine (IL-1ß) and interferon (IFNß), even when a pDNA free from CpG-motifs was encapsulated. The production of the pro-inflammatory cytokines and the DNA sensor-related cytokines is attributed to the combination of vitamin E scaffolds and encapsulated pDNA. The depletion of macrophages by chlodronate-encapsulating liposomes dramatically reduced inflammatory gene expression. Based on the above findings, we conclude that the use of a certain type of non-viral carrier that shows a robust gene expression activity is attended by a risk of eliciting an innate immune response. When a highly hydrophobic derivative of dexamethasone, an anti-inflammatory glucocorticoid compound, was co-loaded to the particle, this inflammatory response was relieved, and gene expression efficiency was enhanced. It is thus concluded that the co-delivery of dexamethasone and pDNA is a promising approach for reducing these risks.

Evaluation of the Microbiological Efficacy of a Single 2-Gram Dose of Extended-Release Azithromycin by Population Pharmacokinetics and Simulation in Japanese Patients with Gonococcal Urethritis.

The objective of this study was to analyze the relationship between the pharmacokinetic (PK)/pharmacodynamic (PD) parameters of a single 2-g dose of extended-release formulation of azithromycin (AZM-SR) and its microbiological efficacy against gonococcal urethritis. Fifty male patients with gonococcal urethritis were enrolled in this study. In 36 patients, the plasma AZM concentrations were measured using liquid chromatography-tandem mass spectrometry, the AZM MIC values for the Neisseria gonorrhoeae isolates were determined, and the microbiological outcomes were assessed. AZM-SR monotherapy eradicated N. gonorrhoeae in 30 (83%) of the 36 patients. AZM MICs ranged from 0.03 to 2 mg/liter. The mean value of the area under the concentration-time curve (AUC), estimated by population PK analysis using a two-compartment model, was 20.8 mg · h/liter. Logistic regression analysis showed that the PK/PD target value required to predict an N. gonorrhoeae eradication rate of ≥95% was a calculated AUC/MIC of ≥59.5. The AUC/MIC value was significantly higher in patients who achieved microbiological cure than in patients who achieved microbiological failure. Monte Carlo simulation using this MIC distribution revealed that the probability that AZM-SR monotherapy would produce an AUC/MIC exceeding the AUC/MIC target of 59.5 was 47%. Furthermore, the MIC distribution for strains isolated in this study was mostly consistent with that for strains currently circulating in Japan. In conclusion, in Japan, AZM-SR monotherapy may not be effective against gonococcal urethritis. Therefore, use of a single 2-g dose of AZM-SR either with or without other antibiotics could be an option to treat gonococcal urethritis if patients are allergic to ceftriaxone and spectinomycin or are diagnosed to be infected with an AZM-sensitive strain.

Analysis of the equine ovarian structure during the first twelve months of life by three-dimensional internal structure microscopy.

A three-dimensional internal structure microscopy (3D-ISM) can clarify the anatomical arrangement of internal structures of equine ovaries. In this study, morphological changes of the equine ovary over the first 12 months of life were investigated by 3D-ISM in 59 fillies and by histological analysis in 2 fillies. The weight and volume of the paired ovaries initially decreased from 0 to 1 months to 2 to 3 months of age and then significantly increased at 8 to 12 months of age. The ovulation fossa was first observed around the 3rd month and became evident after the 6th month. The number of follicles with a diameter of ≥10 mm and the diameter of the largest follicle increased gradually after 6 months of age. On a volume basis, the medulla accounted for nearly 90% of the whole ovary at 0 to 1 months of age, but significantly decreased from 2 to 3 months of age. The volume of the cortex increased progressively after birth and reached approximately 60% of the total volume at 8 to 12 months of age. This significant development of the cortex coincided with the increased number and size of large follicles observed from 6 months of age. These results suggest that the development of the cortex plays a role in the maturation of the follicles and the equine ovary undergoes substantial morphological changes postnatally until puberty.

Artificial oxygen carriers rescue placental hypoxia and improve fetal development in the rat pre-eclampsia model.

Pre-eclampsia affects approximately 5% of all pregnant women and remains a major cause of maternal and fetal morbidity and mortality. The hypertension associated with pre-eclampsia develops during pregnancy and remits after delivery, suggesting that the placenta is the most likely origin of this disease. The pathophysiology involves insufficient trophoblast invasion, resulting in incomplete narrow placental spiral artery remodeling. Placental insufficiency, which limits the maternal-fetal exchange of gas and nutrients, leads to fetal intrauterine growth restriction. In this study, in our attempt to develop a new therapy for pre-eclampsia, we directly rescued placental and fetal hypoxia with nano-scale size artificial oxygen carriers (hemoglobin vesicles). The present study is the first to demonstrate that artificial oxygen carriers successfully treat placental hypoxia, decrease maternal plasma levels of anti-angiogenic proteins and ameliorate fetal growth restriction in the pre-eclampsia rat model.

[A case of subglottic stenosis with bridging granuloma after intubation with double-lumen endotracheal tube].

We present a case of subglottic stenosis with rare bridging granuloma after intubation with double-lumen endotracheal tube. An 81-year-old woman was diagnosed with the lung tumor and scheduled for the thoracoscopic surgery. We induced anesthesia with propofol, remifentanil and rocuronium. A 35 Fr double-lumen intratracheal tube was inserted to the trachea with some resistance, when the tube passed through the glottis. A few days later, she suffered from respiratory discomfort. An otolaryngologist examined her larynx and subglottis. Laryngoscopic examination revealed bridging granuloma leading to tracheal stenosis. Tracheostomy and resection of granuloma were performed, and her symptom improved. If we feel resistance in intubating a double-lumen endotracheal tube in a patient with a history of intubation with a tracheal tube, we should operate gently adjusting the size of the tracheal tube.

Transcriptome tomography for brain analysis in the web-accessible anatomical space.

Increased information on the encoded mammalian genome is expected to facilitate an integrated understanding of complex anatomical structure and function based on the knowledge of gene products. Determination of gene expression-anatomy associations is crucial for this understanding. To elicit the association in the three-dimensional (3D) space, we introduce a novel technique for comprehensive mapping of endogenous gene expression into a web-accessible standard space: Transcriptome Tomography. The technique is based on conjugation of sequential tissue-block sectioning, all fractions of which are used for molecular measurements of gene expression densities, and the block- face imaging, which are used for 3D reconstruction of the fractions. To generate a 3D map, tissues are serially sectioned in each of three orthogonal planes and the expression density data are mapped using a tomographic technique. This rapid and unbiased mapping technique using a relatively small number of original data points allows researchers to create their own expression maps in the broad anatomical context of the space. In the first instance we generated a dataset of 36,000 maps, reconstructed from data of 61 fractions measured with microarray, covering the whole mouse brain (ViBrism: http://vibrism.riken.jp/3dviewer/ex/index.html) in one month. After computational estimation of the mapping accuracy we validated the dataset against existing data with respect to the expression location and density. To demonstrate the relevance of the framework, we showed disease related expression of Huntington's disease gene and Bdnf. Our tomographic approach is applicable to analysis of any biological molecules derived from frozen tissues, organs and whole embryos, and the maps are spatially isotropic and well suited to the analysis in the standard space (e.g. Waxholm Space for brain-atlas databases). This will facilitate research creating and using open-standards for a molecular-based understanding of complex structures; and will contribute to new insights into a broad range of biological and medical questions.

[Influence of intraoperative fentanyl and remifentanil infusion on early postoperative oral intake after general anesthesia].

BACKGROUND: There is no reports on influence of intraoperative fentanyl and remifentanil infusion on ability of oral intake after short stay surgery under general anesthesia. METHODS: Data were collected retrospectively on 497 patients undergoing short stay surgery under general anesthesia with intraoperative remifentanil infusion (R group, n = 273) or without intraoperative remifentanil infusion (NR group, n = 224). The amount of fentanyl used and percentage of patients unable to eat supper were compared. RESULTS: Amount of fentanyl used in patients unable to eat supper were significantly higher than in patients able to eat supper in both NR and R groups. There is no difference in the percentage of patients unable to take supper between group NR and R. Total amount of fentanyl used was significantly lower in group R than in group NR. CONCLUSIONS: With or without the use of remifentanil, greater the amount of fentanyl used, greater the percentage of patients unable to eat. Amount of fentanyl used in R group was significantly less than in group NR; however the difference in percentage of patients unable to eat supper was not observed.

A biomechanical study on burst mechanisms of plant fruit: stress analysis of pericarps before bursting.

Bursting of fruit is a very interesting biomechanical phenomenon because its mechanism is directly related to the plant's reproduction. A plant that produces fruit that bursts powerfully and spreads the seeds widely has the advantage of reproduction without relying on other mechanisms such as transportation of fruit by insects. The structures of many types of fruit have likely been optimized by evolution, although the structure itself appears rather simplistic. Strain energy is stored in each pericarp because of growth deformation, swelling or desiccation just before bursting. Throughout these changes, the mechanical stress of the pericarps is at equilibrium. At the instant of bursting, the stored strain energy is released very rapidly. Quick and wide motion of the pericarps in a certain direction is advantageous for throwing the seed a long distance. The motion and deformation of bursting pericarps depend on their tissue structure and mechanical stress condition just before the burst. We tracked the bursting motion by using a high-speed camera. Then we calculated the pre-burst stress generated in a pericarp of Impatiens by using the finite-element method. The boundary condition obtained by experiments using a high-speed video camera is given, and the stress was calculated using reverse deformation analysis. The stress distribution of the pericarp is effective in causing the pericarp motion to throw the seeds far away.

Expression analysis for inverted effects of serotonin transporter inactivation.

Inactivation of serotonin transporter (HTT) by pharmacologically in the neonate or genetically increases risk for depression in adulthood, whereas pharmacological inhibition of HTT ameliorates symptoms in depressed patients. The differing role of HTT function during early development and in adult brain plasticity in causing or reversing depression remains an unexplained paradox. To address this we profiled the gene expression of adult Htt knockout (Htt KO) mice and HTT inhibitor-treated mice. Inverted profile changes between the two experimental conditions were seen in 30 genes. Consistent results of the upstream regulatory element search and the co-localization search of these genes indicated that the regulation may be executed by Pax5, Pax7 and Gata3, known to be involved in the survival, proliferation, and migration of serotonergic neurons in the developing brain, and these factors are supposed to keep functioning to regulate downstream genes related to serotonin system in the adult brain.

Transgenic frogs expressing the highly fluorescent protein venus under the control of a strong mammalian promoter suitable for monitoring living cells.

To easily monitor living cells and organisms, we have created a transgenic Xenopus line expressing Venus, a brighter variant of yellow fluorescent protein, under the control of the CMV enhancer/chicken beta-actin (CAG) promoter. The established line exhibited high fluorescent intensity not only in most tissues of tadpoles to adult frogs but also in germ cells of both sexes, which enabled three-dimensional imaging of fluorescing organs from images of the serial slices of the transgenic animals. Furthermore, by using this transgenic line, we generated chimeric animals by brain implantation and importantly, we found that the brain grafts survived and expressed Venus in recipients after development, highlighting the boundary between fluorescent and nonfluorescent areas in live animals. Thus, Venus-expressing transgenic frogs, tadpoles, and embryos would facilitate their use in many applications, including the tracing of the fluorescent cells after tissue/organ transplantation.