Simultaneous Imaging and Characterization of Polyunsaturated Fatty Acids, Carotenoids, and Microcrystalline Guanine in Single Aurantiochytrium limacinum Cells with Linear and Nonlinear Raman Microspectroscopy.

Thraustochytrids are heterotrophic marine protists known for their high production capacity of various compounds with health benefits, such as polyunsaturated fatty acids and carotenoids. Although much effort has been focused on developing optimal cultivation methods for efficient microbial production, these high-value compounds and their interrelationships are not well understood at the single-cell level. Here we used spontaneous (linear) Raman and multiplex coherent anti-Stokes Raman scattering (CARS) microspectroscopy to visualize and characterize lipids (e.g., docosahexaenoic acid) and carotenoids (e.g., astaxanthin) accumulated in single living Aurantiochytrium limacinum cells. Spontaneous Raman imaging with the help of multivariate curve resolution-alternating least-squares enabled us to make unambiguous assignments of the molecular components we detected and derive their intracellular distributions separately. Near-IR excited CARS imaging yielded the Raman images at least an order of magnitude faster than spontaneous Raman imaging, with suppressed contributions of carotenoids. As the culture time increased from 2 to 5 days, the lipid amount increased by a factor of ∼7, whereas the carotenoid amount did not change significantly. Furthermore, we observed a highly localized component in A. limacinum cells. This component was found to be mixed crystals of guanine and other purine derivatives. The present study demonstrates the potential of the linear-nonlinear Raman hybrid approach that allows for accurate molecular identification and fast imaging in a label-free manner to link information derived from single cells with strategies for mass culture of useful thraustochytrids.

Observation of the Pockels Effect in Ionic Liquids and Insights into the Length Scale of Potential-Induced Ordering.

Ionic liquids (ILs) under electric fields play essential roles in the electrochemical utilization of ILs. Recently, long-range organization of ILs in the vicinity of charged (and even neutral) surfaces has been revealed, but experimental evidence for such an ordering is still limited and its spatial length scale remains controversial. Here, we use confocal Raman microspectroscopy to investigate the effect of an applied electric potential on the IL 1-butyl-3-methylimidazolium bis(trifluoromethylsulfonyl)imide and its analogues in a space-resolved manner. Much to our surprise, the observed Raman difference spectra of the ILs obtained with and without an applied potential exhibit uniform intensity changes independent of vibrational modes of cations and anions, a finding in sharp contrast with the electric field effects on molecular liquids that we have previously observed. We interpret this unexpected finding in terms of the Pockels effect that occurs as a result of a potential-induced ordering of the IL near an IL-electrode interface. The refractive index changes due to the applied potential are estimated using the experimental Raman intensity changes. The results allow us to deduce that the length scale of the ordering in the ILs is tens to hundreds of nanometers, extending more than would be expected for the electrical double layer but not as far as a micrometer scale.

Multiarm Star-Crosslinked Hydrogel: Polymer Network with Thermoresponsive Free-End Chains Densely Connected to Crosslinking Points.

Soft tissue in biological system is a hydrogel with elaborate structure exhibiting repeatable dynamic function. In order to approach such sophisticated system, precise construction of a designed network with multi-components is desired. This communication presents a novel hydrogel having highly dense stimuli-responsive free-end chains around crosslinking structure. A key molecule is a core-crosslinked star-shaped polymer with multiple thermoresponsive arms, which can be prepared by reversible addition-fragmentation chain transfer polymerization of divinyl crosslinker with poly(N-isopropylacrylamide) (PNIPAAm) macro-chain transfer agent and have a number of unreacted carbon-carbon double bonds in the core. These unreacted double bonds can be utilized as a crosslinker for poly(acrylamide) (PAAm) gel synthesis by free radical polymerization. The obtained gel contains homogeneously dispersed star PNIPAAms as crosslinking points and exhibits thermoresponsive swelling behavior in water depending on the star contents. In particular, the gel with low content of the star crosslinker shows localized responsive behavior with expansion and shrinkage of the star in one molecule. The mechanical properties of the star-crosslinked gel are significantly high compared to the conventional PAAm gels particularly in compressive strength (≈9 MPa). Moreover, the star-crosslinked gel has thermoresponsive mechanical toughening property.

Pulmonary vein thrombosis and cerebral infarction after video-assisted thoracic surgery of the left upper lobe: a case series.

BACKGROUND: Pulmonary vein thrombosis (PVT) and cerebral infarction are rare but critical complications after video-assisted thoracic surgery (VATS). CASE PRESENTATION: We experienced two cases of massive middle cerebral artery infarction after VATS for the left upper lobe. Although the precise source of their embolus was never identified, both cases were clinically suspected PVT. Unfortunately, case 2 died because of progressive cerebral herniation. We decided to perform contrast-enhanced computed tomography routinely after VATS for the left upper lobectomy (VATS-LUL) after these cases. Case 3, a 79-year-old female patient, underwent VATS-LUL for lung cancer. She developed PVT in the stump of the left upper pulmonary vein on postoperative day 4. Anti-coagulation therapy was begun immediately and continued for 3 months. She was free of complications 7 months after the operation. CONCLUSION: PVT and cerebral infarction may occur after VATS-LUL. Appropriate postoperative management is required to recognize PVT and to prevent life-threatening stroke.

Inter- and Intragrain Inhomogeneity in 2D Perovskite Thin Films Revealed by Relative Grain Orientation Imaging Using Low-Frequency Polarized Raman Microspectroscopy.

Two-dimensional (2D) organic-inorganic hybrid lead halide perovskites make up an emerging class of semiconductor materials for optoelectronic applications such as solar cells. The grain structure of polycrystalline 2D perovskites is one of the key factors that dictate their functionality in the devices, but currently available methods for in situ, chemically specific characterization of 2D perovskite grains are scarce. Here we show that ultra-low-frequency polarized Raman microspectroscopy is a facile yet powerful tool for visualizing relative grain orientations within 2D perovskite thin films. We demonstrate this method on the simplest 2D perovskite, (CH3(CH2)3NH3)2PbI4. Hierarchical clustering and detailed band decomposition analysis of the low-frequency polarized Raman imaging data reveal not only relative grain orientations but also intragrain inhomogeneity. We envisage that with high chemical specificity, this method will find broad applications ranging from other 2D perovskites to perovskite-based optoelectronic devices.

Early chest tube removal after surgery for primary spontaneous pneumothorax.

OBJECTIVE: The purpose of this study was to analyze the safety and validity of early chest tube removal after bullectomy for primary spontaneous pneumothorax (PSP). METHODS: Between January 2005 and July 2018, 123 cases of thoracoscopic surgery for PSP were reviewed. The principle procedure was bullectomy accompanied by mechanical and chemical pleurodesis. Approximately 4 h after surgery, we confirmed patients' sufficient recovery from anesthesia and investigated the presence of air leak in a sitting position. Chest tubes were removed on the same day as the surgery if there was no air leak or bloody drainage observed. Postoperative complications and factors that prevented early chest tube removal were analyzed by comparing the early removal group and the remaining tube group. RESULTS: Chest tubes could be removed on the day of the surgery in 105 cases (85.4%). There were 7 cases (5.7%) in which chest tubes could be removed because air leak was not detected after patients' recovery despite intraoperative detection of minor air leak from the resection stump. No patients required chest tube reinsertion during their hospital stay. The mean length of postoperative hospital stay was 1.1 ± 0.5 days. In a logistic regression analysis, surgical history of ipsilateral PSP was independently and significantly associated with the prevention of early chest tube removal. CONCLUSIONS: Chest tube removal on the day of surgery for PSP appears to be safe when air leak examination can be performed after sufficient recovery from anesthesia.

Ultra Cryo-Milling with Liquid Nitrogen and Dry Ice Beads: Characterization of Dry Ice as Milling Beads for Application to Various Drug Compounds.

Ultra cryo-milling using liquid nitrogen (LN2) and dry ice beads has been proposed as a contamination-free milling technique. The morphological change of dry ice beads was visually monitored in LN2 to clarify their production process and cryo-milling process. We found that dry ice pellets, which are starting material of beads and available on the market, immediately disintegrate in LN2, resulting in the spontaneous production of dry ice beads. In addition, the resultant beads maintain their size and shape even under vigorous agitation in LN2, demonstrating that they could play a role of milling media in the milling process. The driving conditions of this cryogenic milling process including beads size were optimized to enhance the milling efficiency. Dry ice beads provided superior milling efficiency compared to original pellets. The milling efficiency increased as the size of the dry ice beads decreased; furthermore, the larger the amount of beads used, the finer the milled particles. Any crystals of three drug compounds were effectively pulverized to the sub- or single-micron range. Cryo-milling with dry ice beads is valuable on pharmaceutical field because it does not contaminate the product with fractured and/or eroded beads.

Hydrogen Bonded Structures of Confined Water Molecules and Electric Field Induced Shift of Their Equilibrium Revealed by IR Electroabsorption Spectroscopy.

Water confined on a nanometer scale plays an essential role in various chemical and biological processes. Confined water molecules are often exposed to electric fields as manifested by those that occur on protein surfaces or in electrical double layers, but the electric field effects on confined water are not fully understood. We used IR electroabsorption (EA) spectroscopy with unprecedented sensitivity to observe electric-field-induced changes in the OH stretching absorption of water (H2O) molecules dissolved in 1,4-dioxane, which constitute a simple model system for confined water. A multivariate curve resolution analysis of the normal IR spectra (without an electric field) of water in 1,4-dioxane at different concentrations indicates the presence of the monomer and dimer of the confined water molecules and equilibrium between them. We find that the IR EA spectrum that is free from the contribution of field-induced molecular reorientation is mainly attributable to a field-induced shift of the equilibrium toward the dimer. This result demonstrates a possible control of the polarity of confined water by simply applying an external electric field and the ability of our method to elucidate how it is achieved.

A new method for electric impedance imaging using an eddy current with a tetrapolar circuit.

A new contactless technique for electrical impedance imaging, using an eddy current managed along with the tetrapolar circuit method, is proposed. The eddy current produced by a magnetic field is superimposed on a constant current that is normally used in the tetrapolar circuit method, and thus is used to control the current distribution in the body. By changing the current distribution, a set of voltage differences is measured with a pair of electrodes. This set of voltage differences is used in the image reconstruction of the resistivity distribution. The least square error minimization method is used in the reconstruction algorithm. The principle of this method is explained theoretically. A backprojection algorithm was used to get 2-D images. Based on this principle, a measurement system was developed and model experiments were conducted with a saline-filled phantom. The estimated shape of each model in the reconstructed image was similar to that of the corresponding model. From the results of these experiments, it is confirmed that the proposed method is applicable to the realization of electrical conductivity imaging.

Venovenous extracorporeal membrane oxygenation support for treatment of bilateral spontaneous pneumothorax.

Bilateral spontaneous pneumothorax is a rare but serious cause of respiratory distress. We treated a 77-year-old male with severe hypoxia caused by bilateral spontaneous pneumothorax using video-assisted thoracoscopic bullectomy assisted by a venovenous extracorporeal membrane oxygenation (ECMO) device. The patient came to the emergency department of our hospital with complaints of cough and dyspnea, and was hospitalized with right-side spontaneous pneumothorax and left-side pneumonia. After 12 days, a chest radiograph was performed to investigate persistent progressive shortness of breath at rest, which demonstrated contralateral pneumothorax. A chest tube was inserted into the left pleural cavity, and surgery was performed for bilateral pneumothorax by video-assisted thoracoscopic surgery (VATS) assisted by venovenous ECMO. Gas exchange was satisfactory throughout the surgical procedure and the postoperative course was uneventful without complications. Venovenous ECMO was effective for facilitation of VATS and reduced the risk of an intra-operative hypoxic condition.

Lung metastasis of thymoma manifesting as myasthenia gravis 12 years after thymomectomy: report of a case.

A 51-year-old woman, who had undergone thymomectomy for asymptomatic noninvasive thymoma 12 years before, was admitted to our hospital with blepharoptosis caused by myasthenia gravis. A chest computed tomogram (CT) showed an abnormal shadow in the right lower lung field and CT-guided needle biopsy revealed findings of a thymoma. We performed extended thymectomy and partial resection of the right lung using three different approaches, via a cervical incision and bilateral video-assisted thoracoscopic surgery. The resected specimen was a lung metastasis of thymoma, and the residual mediastinal tissue showed no sign of malignancy. Because thymoma and post-thymomectomy myasthenia gravis can both recur, we recommend performing extended thymectomy or thymothymectomy, even for patients who are asymptomatic.

Protective effects of preischemic treatment with pioglitazone, a peroxisome proliferator-activated receptor-gamma ligand, on lung ischemia-reperfusion injury in rats.

OBJECTIVES: Lung injury induced by ischemia-reperfusion is the main cause of early graft failure after lung transplantation, which may result from oxygen-free radicals, inflammatory cytokine production, and polymorphonuclear leukocyte accumulation into the interstitium, resulting in severe lung edema. Peroxisome proliferator-activated receptor-gamma (PPAR-gamma) belongs to the nuclear receptor superfamily and has an anti-inflammatory effect by preventing the activation of transcription factors such as nuclear factor-kappaB (NF-kappaB). NF-kappaB regulates the expression of many genes of early response products in the development of acute inflammation. We examined the effects of pioglitazone, a synthetic ligand of PPAR-gamma, against lung ischemia-reperfusion injury in rats. METHODS: The left lungs of male Wistar rats were rendered ischemic for 90 min and then reperfused for 2 h. Treated animals received pioglitazone (10 mg/kg) 2 h before induction of ischemia. Lung injury was quantified in terms of lung microvascular permeability (Evans blue dye extravasation), tissue lipid peroxidation (thiobarbituric acid reactive substances), and tissue polymorphonuclear leukocyte accumulation (myeloperoxidase activity). The tissue concentrations of tumor necrosis factor-alpha (TNF-alpha) and cytokine-induced neutrophil chemoattractant-1 (CINC-1) were also measured. Statistical analyses were performed by one-way analysis of variance, followed by Sheffe's multiple comparison test. RESULTS: The lung vascular permeability in pioglitazone-treated animals was reduced by 55% of the increase of Evans blue dye extravasation relative to control animals (P=0.003). The protective effects of pioglitazone treatment were correlated with the reduction by 79% of the increase of thiobarbituric acid reactive substances (P=0.045) and the reduction by 58% of myeloperoxidase activity increase (P<0.001). The production of TNF-alpha was reduced by 63% of the increase (P<0.001) and the reduction of CINC-1 was 45% (P<0.001). Pioglitazone did not affect the lung in the sham animals. CONCLUSIONS: Pioglitazone treatment before ischemia attenuated lung ischemia-reperfusion injury in rats. The mechanism of these protective effects involves inhibition of the production of proinflammatory cytokines, polymorphonuclear leukocyte accumulation, and tissue lipid peroxidation, resulting in reduced lung edema.

Extrapericardial pulmonary vein clamp technique for pulmonary arteriovenous fistula: report of a case.

The major concern when operating on a patient with a neurologically symptomatic pulmonary arteriovenous fistula (PAVF) is how to prevent a thromboembolic event during surgery. We describe a new technique whereby the extrapericardial pulmonary vein is clamped before transecting the afferent and efferent vessels of the fistula. The potentially pooled clots that can form while manipulating lung are stopped by the clamp. Before the extrapericardial pulmonary vein is declamped, one of the drainage veins is incised halfway and the pooled blood containing the potential clots is completely washed out. We successfully performed segmentectomy using this technique in a 66-year-old man with chronic left hemianopia and a large PAVF in the left anteromedial and lateral basal segments, and no thromboembolic events occurred.

VWRPY motif-dependent and -independent roles of AML1/Runx1 transcription factor in murine hematopoietic development.

AML1/Runx1 is a frequent target of leukemia-associated gene aberration, and it encodes a transcription factor essential for definitive hematopoiesis. We previously reported that the AML1 molecules with trans-activation subdomains retained can rescue in vitro hematopoietic defects of AML1-deficient mouse embryonic stem (ES) cells when expressed by using a knock-in approach. Extending this notion to in vivo conditions, we found that the knock-in ES cell clones with AML1 mutants, which retain trans-activation subdomains but lack C-terminal repression subdomains including the conserved VWRPY motif, contribute to hematopoietic tissues in chimera mice. We also found that germline mice homozygous for the mutated AML1 allele, which lacks the VWRPY motif, exhibit a minimal effect on hematopoietic development, as was observed in control knock-in mice with full-length AML1. On the other hand, reduced cell numbers and deviant CD4 expression were observed during early T-lymphoid ontogeny in the VWRPY-deficient mice, whereas the contribution to the thymus by the corresponding ES cell clones was inadequate. These findings demonstrate that AML1 with its trans-activating subdomains is essential and sufficient for hematopoietic development in the context of the entire mouse. In addition, its trans-repression activity, depending on the C-terminal VWRPY motif, plays a role in early thymocyte development.

Usefulness of new automated cutting needle for tissue-core biopsy of lung nodules under CT fluoroscopic guidance.

OBJECTIVES: The goal of our study was to evaluate the efficacy and safety of a new type of automated cutting needle for tissue-core lung biopsy under real-time CT fluoroscopic guidance. DESIGN: Observational MATERIALS AND METHODS: One hundred ten percutaneous needle tissue-core lung biopsies were performed with the Auto Surecut needle (Create Medic; Yokohama, Japan) under CT fluoroscopic guidance, and the specimens obtained underwent histopathologic evaluation. A final diagnosis was confirmed by independent surgical pathology. RESULTS: The rate of success for the diagnosis for specimens that were adequate for histopathologic analysis was 94.5% (104 of 110 specimens). The sensitivity, specificity, and accuracy in diagnosing malignancy were 95.1%, 100%, and 96.2%, respectively. A specific cell type could be characterized in 95.2% of those 104 lesions (99 lesions; malignant, 76 lesions; benign, 23 lesions). The specific cell type was precisely diagnosed, and was confirmed after surgery in 65 malignant lesions and 23 benign lesions. The biopsy-induced complications encountered were pneumothorax in 34.5% (38 of 110 patients) and hemoptysis in 6.4% (7 of 110 patients). No patient had a serious complication. CONCLUSION: CT fluoroscopy-guided lung biopsy using the Auto Surecut needle provides a high degree of diagnostic accuracy, allows for the specific characterization of lung nodules, and can be performed safely.

Preparation of antibacterial silver-doped silica glass microspheres.

Various types of inorganic substances doped with silver ions have been developed as antibacterial materials, and some have already been commercialized. Colorless and chemically durable materials that slowly release silver ions are, however, still need to be developed. The present authors have previously shown that when a silica glass doped with silver and aluminium ions is prepared using the sol-gel method, the resultant product is colorless, chemically durable, and slowly releases silver ions into water over a long period. The doped silica glass takes a form of microspheres <1 microm in diameter, it is easily mixed with organic polymers, and the mixture can be formed into a thin film or fine fibers, etc. We report on the preparation of silver doped silica glass microspheres having a diameter =1 microm, using the sol-gel method. Initially, tetraethoxysilane was partially prehydrolyzed by water in ethanol, and then aluminium triisopropoxide was added to the solution to form Si-O-Al bonds. Finally, an ammonia solution containing silver nitrate was added to form silica microspheres doped with silver ion together with aluminium ions. The results show monodispersed microspheres 0.4-0.6 microm in diameter were obtained with nominal compositions of Si/Al/Ag = 1/0.01-0.03/0.003-0.03, with a molar ratio of Al/Ag = 1-3.3. The microspheres were colorless, showed a high chemical durability, and slowly released silver ions into water at 37 degrees C. Microspheres with the composition Si/Al/Ag = 1/0.01/0.01 showed excellent antibacterial activity against Escherichia coli. The minimum inhibitory concentration (MIC) of the microspheres was 400, which is less than the MIC value (800) of commercial antibacterial materials.