Development of a C-reactive protein quantification method based on flow rate measurement of an ink solution pushed out by oxygen gas generated by catalase reaction.

A novel determination method for protein biomarkers based on on-chip flow rate measurement was developed using a microchip with organic photodiodes (OPDs). This quantitative method is based on the flow rate measurement of an ink solution pushed out by oxygen gas generated through catalase reaction. The amount of oxygen gas generated in the sample reservoir is dependent on the concentration of the analyte; therefore, the flow rate of the ink solution is also dependent on the concentration of the analyte. The concentration of the analyte can thus be estimated by measurement of the ink solution flow rate. The ink solution flow rate was estimated by measuring the migration time of the ink solution between two points using two OPDs placed below the microchannel. The principle of this method was demonstrated by the measurement of catalase using the microchip. In addition, the developed method was applied to the determination of C-reactive protein (CRP), a biomarker of inflammation, based on a catalase-linked immunosorbent assay (C-LISA). The limit of detection for CRP was 0.20 µg/mL. The method was also applied to the determination of CRP in human serum, and the quantitative values obtained by this method were in excellent agreement with those obtained by the conventional enzyme-linked immunosorbent assay (ELISA) method. The developed method does not require a photodetector with high sensitivity and is thus capable of downsizing; therefore, this will be useful for on-site analyses such as point-of-care testing and field measurements.

Phototunable magnetism in copper octacyanomolybdate.

We introduce copper molybdenum cyanides of general formula Cu2[Mo(CN)8] · nH2O, which can serve as optofunctional magnetic devices. Their ground states generally stay paramagnetic down to temperatures of the K order but exhibit a spontaneous magnetization upon photoirradiation usually below a few tens of K. To interest us still further, such a ferromagnetic stateinduced by blue-laser irradiation is demagnetized step by step through further application of red or near-infrared laser pulses. We solve this intriguing photomagnetism. The ground-state properties are fully revealed by means of a group-theoretical technique. Taking account of experimental observations, we simulate applying pump laser pulses to a likely ground state and successfully reproduce both the magnetization and demagnetization dynamics. We monitor the photorelaxation process through angle-resolved photoemission spectroscopy. Electrons are fully itinerant in any of the photoinduced steady states, forming a striking contrast to the initial equilibrium state of atomic aspect. The fully demagnetized final steady state looks completely different from the initial paramagnetism but bears good analogy to one of the possible ground states available with the Coulomb repulsion on Cu sites suppressed.

Development of small-sized fluorescence detector for pipette tip-based biosensor for on-site diagnosis.

A small-sized fluorescence detector (referred to as a pipette tip [PT]-reader) was developed for a pipette tip-based biosensor. The PT-reader allows us to measure the fluorescence intensity of a solution in a truncated cone-shaped pipette tip with only the tip inserted into the PT-reader. A pipette holder made from a mixture of polydimethylsiloxane (PDMS) and carbon black was capable of the rigorous position arrangement of a truncated cone shaped-pipette tip and the prevention of stray light. The detection performance of the PT-reader was evaluated by measurement of resorufin. The limit of detection (LOD; 3σ) and the relative standard deviation (RSD, n = 4) were estimated to be 0.46 µM and 0.47-4.1%, respectively. This performance was comparable to that of a desktop-type fluorescence microplate reader. In addition, the PT-reader was applied to the quantification of immunoglobulin A (IgA), and the LOD (3σ) of IgA was estimated to be 1.0 ng/mL. The quantitation values of IgA in human saliva obtained by the PT-based enzyme-linked immunosorbent assay (PT-ELISA) were in agreement with those obtained by conventional ELISA. The PT-reader is expected to be useful for low-cost and user-friendly measurements, and the technique of device development proposed in this study will contribute to the progress of on-site medical diagnosis.

Bottom-up realization of a porous metal-organic nanotubular assembly.

Nanotubes are generally prepared from their constituent elements at high temperatures, and thus it is difficult to control their size, shape and electronic states. One useful approach for synthesizing well-defined nanostructures involves the use of building blocks such as metal ions and organic molecules. Here, we show the successful creation of an assembly of infinite square prism-shaped metal-organic nanotubes obtained from the simple polymerization of a square-shaped metal-organic frame. The constituent nanotube has a one-dimensional (1D) channel with a window size of 5.9×5.9 Å(2), and can adsorb water (H(2)O) and alcohol vapours, whereas N(2) and CO(2) do not adhere. It consists of four 1D covalent chains that constitute a unique electronic structure of 'charge-density wave (CDW) quartets' on crystallization. Moreover, exchanging structural components and guest molecules enables us to control its semiconductive bandgap. These findings demonstrate the possibility of bottom-up construction of new porous nanotubes, where their degrees of freedom in both pore space and framework can be used.

Asian sand dust enhances murine lung inflammation caused by Klebsiella pneumoniae.

Inhaling concomitants from Asian sand dust (ASD) may result in exacerbation of pneumonia by the pathogen. The exacerbating effect of ASD on pneumonia induced by Klebsiella pneumoniae (KP) was investigated in ICR mice. The organic substances adsorbed onto ASD collected from the atmosphere of Iki-island in Japan were excluded by heat treatment at 360°C for 30min. ICR mice were instilled intratracheally with ASD at doses of 0.05mg or 0.2mg/mouse four times at 2-week intervals (total dose of 0.2mg or 0.8mg/mouse) and were administrated with ASD in the presence or absence of KP at the last intratracheal instillation. Pathologically, ASD caused exacerbation of pneumonia by KP as shown by increased inflammatory cells within the bronchiolar and the alveolar compartments. ASD enhanced the neutrophil number dose dependently as well as the expression of cytokines (IL-1ß, IL-6, IL-12, IFN-γ, TNF-α) and chemokines (KC, MCP-1, MIP-1α) related to KP in BALF. In an in vitro study using RAW264.7 cells, combined treatment of ASD and KP increased gene expression of IL-1ß, IL-6, IFN-ß, KC, MCP-1, and MIP-1α. The same treatment tended to increase the protein level of IL-1ß, TNF-α and MCP-1 in a culture medium compared to each treatment alone. The combined treatment tended to increase the gene expression of Toll-like receptor 2 (TLR2), and NALP3, ASC and caspase-1 compared with KP alone. These results suggest that the exacerbation of pneumonia by ASD+KP was due to the enhanced production of pro-inflammatory mediators via activation of TLR2 and NALP3 inflammasome pathways in alveolar macrophages.

Texture Management for Glossy Objects Using Tone Mapping.

In this paper, we proposed a method for matching the color and glossiness of an object between different displays by using tone mapping. Since displays have their own characteristics, such as maximum luminance and gamma characteristics, the color and glossiness of an object when displayed differs from one display to another. The color can be corrected by conventional color matching methods, but the glossiness, which greatly changes the impression of an object, needs to be corrected. Our practical challenge was to use tone mapping to correct the high-luminance part, also referred to as the glossy part, which cannot be fully corrected by color matching. Therefore, we performed color matching and tone mapping using high dynamic range images, which can record a wider range of luminance information as input. In addition, we varied the parameters of the tone-mapping function and the threshold at which the function was applied to study the effect on the object's appearance. We conducted a subjective evaluation experiment using the series category method on glossy-corrected images generated by applying various functions to each display. As a result, we found that the differences in glossiness between displays could be corrected by selecting the optimal function for each display.

Fabrication of a Human Skin Mockup with a Multilayered Concentration Map of Pigment Components Using a UV Printer.

In this paper, we propose a pipeline that reproduces human skin mockups using a UV printer by obtaining the spatial concentration map of pigments from an RGB image of human skin. The pigment concentration distributions were obtained by a separating method of skin pigment components with independent component analysis from the skin image. This method can extract the concentration of melanin and hemoglobin components, which are the main pigments that make up skin tone. Based on this concentration, we developed a procedure to reproduce a skin mockup with a multi-layered structure that is determined by mapping the absorbance of melanin and hemoglobin to CMYK (Cyan, Magenta, Yellow, Black) subtractive color mixing. In our proposed method, the multi-layered structure with different pigments in each layer contributes greatly to the accurate reproduction of skin tones. We use a UV printer because the printer is capable of layered fabrication by using UV-curable inks. As the result, subjective evaluation showed that the artificial skin reproduced by our method has a more skin-like appearance than that produced using conventional printing.

Potent and Selective PTDSS1 Inhibitors Induce Collateral Lethality in Cancers with PTDSS2 Deletion.

SIGNIFICANCE: This study identifies a specific dependency on PTDSS1 for phosphatidylserine synthesis following PTDSS2 deletion and introduces novel PTDSS1 inhibitors as a therapeutic option to induce collateral lethality in cancer with PTDSS2 loss.

Novel Antibody Exerts Antitumor Effect through Downregulation of CD147 and Activation of Multiple Stress Signals.

CD147 is an immunoglobulin-like receptor that is highly expressed in various cancers and involved in the growth, metastasis, and activation of inflammatory pathways via interactions with various functional molecules, such as integrins, CD44, and monocarboxylate transporters. Through screening of CD147-targeting antibodies with antitumor efficacy, we discovered a novel rat monoclonal antibody #147D. This humanized IgG4-formatted antibody, h4#147D, showed potent antitumor efficacy in xenograft mouse models harboring the human PDAC cell line MIA PaCa-2, HCC cell line Hep G2, and CML cell line KU812, which featured low sensitivity to the corresponding standard-of-care drugs (gemcitabine, sorafenib, and imatinib, respectively). An analysis of tumor cells derived from MIA PaCa-2 xenograft mice treated with h4#147D revealed that cell surface expression of CD147 and its binding partners, including CD44 and integrin α3ß1/α6ß1, was significantly reduced by h4#147D. Inhibition of focal adhesion kinase (FAK), activation of multiple stress responsible signal proteins such as c-JunN-terminal kinase (JNK) and mitogen-activated protein kinase p38 (p38MAPK), and expression of SMAD4, as well as activation of caspase-3 were obviously observed in the tumor cells, suggesting that h4#147D induced tumor shrinkage by inducing multiple stress responsible signals. These results suggest that the anti-CD147 antibody h4#147D offers promise as a new antibody drug candidate.

Development of an automated transportable continuous system to measure the total alkalinity of seawater.

Anthropogenic CO2 emissions are contributing to global warming and ocean acidification. Rapid and accurate measurements of seawater carbonate chemistry are critical to understand current changes in the ocean and to predict future effects of such changes on marine organisms and ecosystems. Total alkalinity (AT) measurements can be used to directly determine the calcification rate, but they are time-consuming and require large sample volumes. Herein, we describe an automated and transportable flow-through system that can conduct continuous AT measurement using an ion sensitive field effect transistor (ISFET) - Ag/AgCl sensor and three different reference materials. The response time, stability, and uncertainty of our system were evaluated by comparing AT values of calibrated reference materials to those calculated by our system. Our system requires only small amounts of seawater (<10 mL) and a short time per sample (<5 min) to produce results with a relative uncertainty of less than 0.1% (approx. 2.2 µmol kg-1). This system is expected to facilitate easy and rapid in-situ measurement of AT. Continuous AT measurements would enable us to determine short-term calcification responses to changes in light or temperature and improve our understanding of the metabolic mechanisms of creatures such as corals.

Strain differences influence N-methyl-D-aspartate receptor subunit gene expression in the olfactory bulb of an allergic mouse model following toluene exposure.

OBJECTIVE: Genetic variation is a causative factor in differing sensitivities to environmental chemicals. The present study explored whether differences in mouse strains influence N-methyl-D-aspartate (NMDA) receptor expression in the olfactory bulb after low-level toluene exposure. METHODS: Ten-week-old male C3H/HeN (H-2(k)), BALB/c (H-2(d)) and C57BL/10 (H-2(b)) mice were exposed to 0, 5, 50 or 500 ppm of toluene for 6 h per day, 5 days per week for 6 weeks. Because individuals with allergic disease are more susceptible to volatile organic compound exposure, the animals of each strain were divided into 2 main groups, a non-allergy (NAG) group and an allergy (AG) group. The AG groups were stimulated with ovalbumin before toluene exposure. The mRNA levels of NMDA receptor subunits (NR1, NR2A and NR2B) in the olfactory bulbs of the NAG and AG groups were examined using real-time RT-PCR. RESULTS: In C3H/HeN mice, the expression levels of NR1 and NR2B mRNA decreased significantly in the AG group exposed to 500 ppm of toluene; in the NAG group, however, the NR2A mRNA level increased significantly in mice exposed to 50 ppm while the NR2A and NR2B mRNA levels decreased significantly in mice exposed to 500 ppm of toluene. No significant changes were observed in the NAG groups of BALB/c or C57BL/10 mice. However, in the BALB/c mice, the mRNA levels of NR1, NR2A and NR2B decreased significantly in the AG group exposed to 500 ppm of toluene. CONCLUSION: Mammalian strain differences in NMDA receptor expression after allergic stimulation can be observed in the mouse olfactory bulb after toluene exposure.

Different sensitivity in expression of transcription factor mRNAs in congenic mice following exposure to low-level toluene.

To investigate the relationship between sensitivity to toluene exposure and genetic background, male congenic mice, C57BL/10 (H-2(b)) and B10.BR/Sg (H-2(k)) were exposed to 0, 5, and 50 ppm toluene for 6 h per day, 5 days per week for 6 weeks. Groups of mice were injected with ovalbumin (OVA) intraperitoneally before starting exposure schedule and these mice were then challenged with aerosolized OVA as a booster. Following 24 h of the last exposure, the spleens were collected. We examined spleen cell proliferation using DNA synthesis and T-helper 1/2-related transcription factor genes in spleen of two congenic mice using real-time reverse transcriptase-polymerase chain reaction. Although lipopolysaccharide-induced (LPS-induced) cell proliferation was significantly increased in 50 ppm toluene-exposed, nonimmunized B10.BR mice but not in C57BL/10 mice, exposure to 50 ppm toluene significantly decreased LPS-induced cell proliferation in immunized B10.BR mice. The expression of transcription factor forkhead box P3, signal transducers and activators of transcription (STAT)5 and STAT6 mRNAs was significantly increased in spleen from 5 ppm toluene-exposed, OVA-immunized B10.BR mice, but not in those of C57BL/10 mice. Although there may be other differences unrelated to H-2 locus between the congenic mice of the same background, the findings of the present study strongly suggest a possible role of H-2 locus in the toluene-induced immune disturbance.

Strain differences in extracellular amino acid neurotransmitter levels in the hippocampi of major histocompatibility complex congenic mice in response to toluene exposure.

OBJECTIVE: The aim of the present study was to investigate the possible role of the major histocompatibility complex locus in neurotransmitter systems in the mouse hippocampus following toluene exposure. METHODS: We compared the changes in toluene-induced extracellular amino acid neurotransmitter levels in the hippocampi of 2 strains of male congenic mice, C57BL/10 (H-2(b)) and B10.BR/Sg (H-2(k)). In vivo microdialysis was performed in each freely moving mouse after a single intraperitoneal injection of toluene (300 mg/kg), and neurotransmitters in the hippocampal microdialysates were measured using high-performance liquid chromatography. RESULTS: The basal extracelluar glutamate and glycine levels in the hippocampi of the C57BL mice were significantly higher than those in the B10.BR mice. However, the basal extracellular taurine levels in the hippocampi of the C57BL mice were significantly lower than those in the B10.BR mice. Although no changes in the glutamate levels were observed after toluene injection in either strain, the glycine levels increased significantly after toluene injection in the C57BL mice. On the other hand, significantly lower taurine levels were observed after toluene injection in both strains of mice. CONCLUSIONS: Our data demonstrate the existence of a relationship between H-2 haplotypes and hippocampal neurotransmitter levels in mice.

Suppression of Th1- and Th2-type immune responses in infant mouse spleen after prenatal and postnatal exposure to low-level toluene and peptidoglycan.

The aim of the present study was to investigate the effect of low-level concentrations, under the occupational acceptable limits, of toluene exposure and peptidoglycan (PGN) stimulation on Th1/Th2 immunity in infant mice. Pregnant BALB/c mice and their offspring were exposed to low-level toluene inhalation (0, 5, and 50 ppm) for 4 wk (from the late prenatal stage to early postnatal stage) in a whole-body exposure chamber. Some of the pregnant mice and their offspring were stimulated with PGN during toluene exposure. We measured total immunoglobulins of different subclasses in plasma, and production and expression level of cytokines in the lung and spleen, and transcription factors related to Th1/Th2 immunity in the spleen of infant (3 wk old) mice. Exposure of mice to 5 or 50 ppm toluene resulted in increased immunoglobulin (Ig) G1 and decreased IgG2a and IgE antibodies in the plasma; significantly decreased T-bet, GATA-3, and Foxp3 mRNA in the spleen; and a tendency toward decreased interferon (IFN)-gamma mRNA in spleen. Exposure of mice to low-level toluene together with PGN stimulation resulted in decreased IgG1 as well as IgG2a antibodies in the plasma and Foxp3 mRNA in spleen as compared with control or PGN-treated mice. These findings suggest that low-level toluene exposure and PGN stimulation from the late prenatal to early postnatal stage suppressed the splenic parameter related to Th1/Th2 immunity in infant mice.

Extracellular glutamate level and NMDA receptor subunit expression in mouse olfactory bulb following nanoparticle-rich diesel exhaust exposure.

In this present study, we aimed to investigate the extracellular glutamate level and memory function-related gene expression in the mouse olfactory bulb after exposure of the animals to nanoparticle-rich diesel exhaust (NRDE) with or without bacterial cell wall component. Lipoteichoic acid (LTA), a cell wall component derived from Staphylococcus aureus, was used to induce systemic inflammation. Male BALB/c mice were exposed to clean air (particle concentration, 4.58 microg/m(3)) or NRDE (148.86 microg/m(3)) 5 h per day on 5 consecutive days of the week for 4 wk with or without weekly intraperitoneal injection of LTA. We examined the extracellular glutamate levels in the olfactory bulb using in vivo microdialysis and high-performance liquid chromatography assay. Then, we collected the olfactory bulb to examine the expression of N-methyl-D-aspartate (NMDA) receptor subunits (NR1, NR2A, and NR2B) and calcium/calmodulin-dependent protein kinase (CaMK) IV and cyclic AMP response element binding protein (CREB)-1 using real-time reverse-transcription polymerase chain reaction (RT-PCR). NRDE and/or LTA caused significantly increased extracellular glutamate levels in the olfactory bulb of mice. Moreover, the exposure of mice to NRDE upregulates NR1, NR2A, NR2B, and CaMKIV mRNAs in the olfactory bulb, while LTA upregulates only NR2B and CREB1 mRNAs. These findings suggest that NRDE and LTA cause glutamate-induced neurotoxicity separately and accompanied by changes in the expression of NMDA receptor subunits and related kinase and transcription factor in the mouse olfactory bulb. This is the first study to show the correlation between glutamate toxicity and memory function-related gene expressions in the mouse olfactory bulb following exposure to NRDE.

Role of CD4(+) T cells in the modulation of neurotrophin production in mice exposed to low-level toluene.

To investigate the role of CD4(+) T cells in neurotrophin production following toluene exposure, male C3H mice were exposed to filtered air (control) or 9 ppm of toluene in a nose-only exposure chamber for 30 min on 3 consecutive days followed by weekly sessions for 4 weeks. All the mice were immunized with ovalbumin and some groups of mice were treated with anti-CD4 antibody. BDNF content in BAL fluid and NGF content in plasma were significantly increased in toluene-exposed mice. However, treatment with anti-CD4 mAb completely abrogated these effects. These findings suggest that the CD4(+) T cells may be involved in the toluene-induced modulation of neurotrophin production.

Establishment of a mouse model to assess brain neurotransmitter level and learning performance simultaneously following toxic chemical exposure: using in vivo microdialysis and schedule-controlled operant behavior.

Most of the volatile organic compounds used in industrial and household products are reported to be toxic. While the effects of toxic chemicals on pulmonary and other systemic functions have been studied substantially, little is known about their effects on higher brain functions, particularly on the events in learning performance. From the particular perspective of environmental health, it is necessary to examine the toxic effects of environmental chemicals on higher brain functions. The aim of the present study is to establish a mouse model for simultaneous assessment of the effect of toluene (150 mg/kg, i.p.) on extracellular glutamate level in the hippocampus, using in vivo microdialysis and learning performance with a procedure of schedule-controlled operant behavior (SCOB). Different patterns of hippocampal glutamate level were observed in control and toluene-treated mice during learning performance. During learning performance in an operant chamber, control mice showed a saturated and persistent increased glutamate level with good learning performance, while toluene-treated mice showed a decreased glutamate level with poor learning performance. This is the first time to establish a new method to assess the effect of environmental chemical exposure on the brain neurotransmitter level during learning performance in an animal model.

Spatial learning and memory function-related gene expression in the hippocampus of mouse exposed to nanoparticle-rich diesel exhaust.

Diesel exhaust particles are a major constituent of ambient particulate matter, and most particles emitted directly from diesel exhaust are smaller than 1microm in diameter. Recently, the toxicity of diesel engine-derived nanoparticles has come to be recognized as an emerging social issue. In the present study, we investigated spatial learning ability and memory function-related gene expressions in mouse hippocampus after the exposure of animals to nanoparticle-rich diesel exhaust (NRDE) with or without a bacterial cell wall component. Lipoteichoic acid (LTA), a cell wall component derived from Staphylococcus aureus, was used to induce systemic inflammation. Male BALB/c mice were exposed to clean air (particle concentration, 4.58microg/m(3)) or NRDE (148.86microg/m(3)) for 5h per day on 5 days of the week for 4 weeks in an exposure chamber, with or without the weekly intraperitoneal injection of LTA. On the day after the final day of exposure, we used a Morris water maze apparatus to examine the ability of the animals to perform a spatial learning task. After the completion of the test, the animals were sacrificed and the hippocampus was collected from each mouse; the expressions of NMDA receptor subunits (NR1, NR2A and NR2B), proinflammatory cytokines (IL-1beta and TNF-alpha) and the oxidative stress marker heme oxygenase 1 were then investigated using real-time RT-PCR. In the Morris water maze task, NRDE/LTA (+) group took a longer time to reach the hidden platform than clear air/LTA (-) group. However, NRDE exposure alone did not affect it. The relative mRNA levels of the NMDA receptor subunits and proinflammatory cytokines were higher in hippocampus of NRDE/LTA (+) group compared to clear air/LTA (-) group. These results indicate that co-exposure of NRDE and LTA could affect spatial learning and memory function-related gene expressions in mouse hippocampus.

[Biodistribution of ultrafine particles of titanium dioxide by intratracheal administration to mice].

TiO2 ultrafine particles are used as photo-catalysis. When ultrafine particles are exposed to hosts, they are invaded in alveolar, transferred to organs through blood vessels and may express biological effects. We administered TiO2 ultra-fine particles (5 nm, 100 nm) intratracheally to mice, and collected whole blood and removed organs (liver, lung, kidney, spleen and brain) after 1, 4 and 24 hours. The quantity of Ti in the blood and these organs was analyzed by PIXE (Particle Induced X-Ray Emission) or ICP/MS (Inductively Coupled Plasma-Mass Spectrometry). Compared to control mice, the quantity of Ti in the exposed mice was not different. Consequently, we observed the solution of dissolved TiO2 ultrafine particles by Scanning Electron Microscope, and observed the particles which aggregated. That diameter was about 1 microm. We concluded that the particles had aggregated before administration to mice, so they didn't invade the blood vessels or organs from the pulmonary alveolus in the lung.

Organocatalytic Stereoselective Cyclic Polylactide Synthesis in Supercritical Carbon Dioxide under Plasticizing Conditions.

Cyclic polylactide (cPLA) is a structural isomer of linear polylactide (PLA) although it possesses unique functionalities in comparison to its linear counterpart. Hitherto, the control of stereochemical purity in conventional cPLA synthesis has not been achieved. In this study, highly stereochemically pure cPLA was synthesized in the absence of a metal catalyst and organic solvent, which required high consumption of the residual monomer. The synthesis was conducted in supercritical carbon dioxide under CO2 plasticizing polymerization conditions in the presence of an organocatalyst and thiourea additives. In comparison with the stereocomplexes synthesized through conventional methods, cPLA from l-lactide (cPLLA) and cPLA from d-lactide (cPDLA) were synthesized with higher stereochemical purity and improved thermal stability. Moreover, the method presented herein is environmentally friendly and thus, applicable on an industrial level.