Simple solid-phase colorimetry for trace Cr(VI) by combination of complexation with diphenylcarbazide and ion-pair solid-phase extraction with sedimentable dispersed particulates.

A highly sensitive and simple solid-phase colorimetry for Cr(VI) was proposed. It was based on the ion-pair solid-phase extraction of Cr-diphenylcarbazide (DPC) complex with sedimentable dispersed particulates. The concentration of Cr(VI) was measured from the color tones obtained by image analysis of the photo of sediment. Various conditions, e.g., material and amounts of adsorbent particulates, chemical properties and concentration of counter ions, and pH, were optimized for the formation and quantitative extraction of the complex. In the recommended procedure, 1 mL of sample was put into a 1.5 mL microtube where powder form adsorbent and reagents, i.e., XAD-7HP particles, DPC, sodium dodecyl sulfate, amido sulfuric acid, and sodium chloride had been packed. The analytical operation was completed within 5 min by gently shaking the microtube and allowing it to stand until enough amounts of particulates were deposited to take a picture. Chromium (VI) up to 2.0 ppm was determined, and the detection limit was 0.0034 ppm. The sensitivity was enough to determine Cr(VI) at lower concentrations than the water quality of standard (0.02 ppm). This method was successfully applied to the analysis of simulated industrial wastewater samples. The stoichiometry of the extracted chemical species was also investigated by applying the same equilibrium model as the ion-pair solvent extraction.

Lossy mode resonance fiber-optic sensor based on ZnO particles fabricated by chemical bath deposition.

We present a lossy mode resonance (LMR) sensor fabricated by chemical bath deposition (CBD) using a U-shaped optical fiber with an exposed core. The ZnO particles that generate LMR were prepared by a very costly method in three steps via permanganate activation and the deposition of ZnO on the fiber core using CBD. The process of deposition was monitored in real time through the optical fiber, and a clear absorption spectrum with an LMR peak was obtained. The surface of the sensor with absorbance reaching 1.0 was covered with nano- to submicron particles of ZnO. The refractive index (RI) sensitivity of the sensor was measured using sucrose solution and was found to increase as the amounts of ZnO on the sensor increased, reaching 23 Abs/RI unit (RIU). The RI resolutions of the sensors with absorbance reaching 0.40, 0.65, and 1.0 during CBD were determined as ΔRI = 0.000060, 0.00017, and 0.00018, respectively, with a 99.7% confidence interval for the RI. Pretreatment during CBD was found to dramatically affect the fabrication of LMR sensors owing to their size and occupancy of deposited ZnO particles, the effects of which can be observed in real time using fiber optics.

Colorimetric analysis based on solid-phase extraction with sedimentable dispersed particulates: demonstration of concept and application for on-site environmental water analysis.

A novel simple and functional colorimetric methodology for on-site environmental water analysis was proposed. This method combines coloration of the analyte and extraction of the colored species on dispersed particulates during their sedimentation in the same container. The whole analysis can be performed within 15 min by comprising the addition of 1 mL of sample solution into a 1.5-mL microtube containing the powders of coloring reagents and the sedimentable fine particulates as an adsorbent. The analyte is determined by comparing the sediment color with the standard color by visual inspection or the color information of the photo image. The potential of this methodology was demonstrated through developing colorimetry for Fe2+ with o-phenanthroline, NO2- by azo-dye formation, HCHO by the MBTH method, and PO43- by the 4-aminoantipyrine method based on the enzyme reactions. The material, size, amount of the adsorbent particles, and other conditions were optimized for each analytes. The advantages of the methodology were as follows: high sensitivity, easy controllability of the sensitivity over the wide range by the amount, size, and material of the particulates, lower interference from the colored matrix components due to obtaining the color data from not the aqueous phase but the sedimented particulates, and acceleration of the color development rate by the particulates as seen in NO2- determination as consequence shorten the operation time. A simple device equipped with twin cells was proposed for on-site analysis which contains two successive different coloring operations. The developed methods were successfully applied to the environmental water samples with the good agreement of the results with those by the usual instrumental methods.

U-shaped plastic optical fiber sensor for scale deposition in hot spring water.

Fiber optic sensors for monitoring scale deposition in geothermal brine and hot spring water should be safe, easily fabricated, and readily disposable. These desired features already have been enhanced in plastic optical fibers (POFs) and U-shaped sensors for other applications. The present work reports a U-shaped POF sensor for CaCO3 scale deposition. The sensors were easily fabricated by thermally bending the bulk POF without removing the cladding. At the bend, the percentage of total internal reflection between the water and the POF surface is affected by the high refractive index of the CaCO3 deposit. The optical responses of the U-shaped sensor to CaCO3 formation were investigated in a mixture of calcium chloride dehydrate and sodium hydrogen carbonate using a white-light source and a spectroscopic detector. The sensor was responsive to CaCO3 formation on the sensor surface and was especially sensitive at small bending radii. The sensitivity was further enhanced by increasing the number of bends. Finally, the U-shaped POF sensor was applied to the monitoring of CaCO3 scale deposition in hot spring water sampled at Matsushiro, Japan.

Electrochemical Long Period Fiber Grating Sensing for Electroactive Species.

We present an electrochemical long period fiber grating (LPFG) sensor for electroactive species with an optically transparent electrode. The sensor was fabricated by coating indium tin oxide onto the surface of LPFG using a polygonal barrel-sputtering method. LPFG was produced by an electric arc-induced technique. The sensing is based on change in the detection of electron density on the electrode surface during potential application and its reduction by electrochemical redox of analytes. Four typical electroactive species of methylene blue, hexaammineruthenium(III), ferrocyanide, and ferrocenedimethanol were used to investigate the sensor performance. The concentrations of analytes were determined by the modulation of the potential as the change in transmittance around the resonance band of LPFG. The sensitivity of the sensor, particularly to methylene blue, was high, and the sensor responded to a wide concentration range of 0.001 mM to 1 mM.

Evanescent-Wave Fiber Optic Sensing of the Anionic Dye Uranine Based on Ion Association Extraction.

Herein, we propose an evanescent-wave fiber optic sensing technique for the anionic dye uranine based on ion association extraction. The sensor was prepared by removing a section of the cladding from a multimode fiber and hydrophobization of the exposed core surface. Uranine was extracted in association along with hexadecyltrimethylammonium (CTA) ion onto the fiber surface and detected via absorption of the evanescent wave generated on the surface of the exposed fiber core. The effect of CTA+ concentration added for ion association was investigated, revealing that the absorbance of uranine increased with increasing CTA+ concentration. A change in the sensor response as a function of the added uranine concentration was clearly observed. The extraction data were analyzed using a distribution equilibrium model and a Freundlich isotherm. The uranine concentration in the evanescent field of the fiber optic was up to 54 times higher than that in the bulk solution, and the limit of detection (3σ) for uranine was found to be 1.3 nM.

Organic Ion-associate Phase Extraction/Back-microextraction for the Preconcentration and Determination of Lithium Using 2,2,6,6-Tetramethyl-3,5-heptanedione by Liquid Electrode Plasma Atomic Emission Spectrometry and GF-AAS in Environmental Water.

We developed an ion-associate phase (IAP)-extraction/acid back-extraction system for the preconcentration and atomic spectrometric determination of lithium trace amounts in water. The chelating reagent for lithium also works as a constituent of the extraction phase. The lithium in a 10 mL sample solution was converted through a chelate complex reaction with 2,2,6,6-tetramethyl-3,5-heptanedione (HDPM). The addition of a benzyldimethyltetradecylammonium ion caused the formation of IAP suspension in the solution. Centrifugation of the solution led to the isolation of a liquid organic phase and the lithium complex was extracted as the upper phase from the centrifuge tube. After the aqueous phase was removed, lithium was back-extracted with a 400 µL nitric acid solution from the IAP. The acid phase was measured using liquid-electrode-plasma atomic-emission-spectrometry (LEP-AES) or graphite-furnace atomic-absorption spectroscopy (GF-AAS). The detection limits were 0.02 mg/L for LEP-AES and 0.02 µg/L for GF-AAS. This system was applied to the determination of environmental water. The HDPM in the organic phase was reusable.

Investigation of the effects of electromagnetic field treatment of hot spring water for scale inhibition using a fibre optic sensor.

Treatment with an electromagnetic field, one of the potential techniques to inhibit scale deposition from water, has the advantage of not requiring the addition of any chemicals. Field tests using a fibre optic sensor were conducted to evaluate the effect that the treatment of hot spring water in Matsushiro, Japan with an electromagnetic field had on calcium carbonate scale formation. The optical response to scale deposition recorded by the fibre optic sensor decreased as a consequence of the application of an electromagnetic field, and the effectiveness of scale formation inhibition depended on the frequency of the electromagnetic field. This evidence was compared with results from changes in scale mass measured using the quartz crystal microbalance (QCM) method. Mass increases of the scale formed on the quartz crystal surface in hot spring water were inhibited by electromagnetic field treatment. These results were verified performing a column flow test, whereby the flow rate of hot spring water through a column was measured.

Micro-organic Ion-associate Phase Extraction/micro-volume Back-extraction for the Preconcentration and GF-AAS Determination of Cadmium, Nickel and Lead in Environmental Water.

Micro-organic ion-associate phase (IAP) extraction was combined with a micro-volume back-extraction (MVBE) to reduce coexisting components and viscosity in the concentrates. Heavy metals were converted into a complex with 2-(5-bromo-2-pyridylazo)-5-(N-propyl-N-sulfopropylamino)phenol in a 40-mL sample solution, and were extracted into ion associates. After centrifugation and discarding the aqueous phase, trace metals were stripped from IAP into a nitric acid solution, followed by GF-AAS determination. Only one vessel was required for 400-fold enrichment. The detection limits (3σb) for Cd, Ni, and Pb were 0.6, 3.7, and 0.8 ng/L, respectively. This method was applied in recovery tests in seawater.

Simultaneous Multiselective Spectroelectrochemical Fiber-Optic Sensor: Sensing with an Optically Transparent Electrode.

We present a spectroelectrochemical fiber-optic sensor with an optically transparent electrode. The sensor was fabricated by coating indium tin oxide (ITO) onto the surface of fiber-optic core chips using a polygonal barrel-sputtering method. The ITO-coated fiber-optic probe can be simply and cheaply mass-produced and used as a disposable probe. The sensing is based on changes in an attenuated total reflection signal accompanying the electrochemical oxidation-reduction of an analyte at the electrode. The properties of an ITO-coated fiber-optic probe as an optically transparent electrode were investigated for varying thicknesses of ITO. The sensor responses were successfully enhanced with an additional level of selectivity via an electrostatically adsorbed, self-assembled monolayer, which comprised a polyanion and polycation.

Development of an Attenuated Total Reflection Based Fiber-Optic Sensor for Real-time Sensing of Biofilm Formation.

A fiber-optic sensor capable of real-time monitoring of biofilm formation in water was developed. The sensor can be easily fabricated by removing the cladding of a multimode fiber optic to expose the core. The sensing action is based on the penetration of an evanescent wave through a biofilm formed on the surface of the exposed fiber core during total internal reflection. The proposed setup can be used to analyze the transmittance response over a wide wavelength range using a white-light source and a spectroscopy detector. The change in transmittance with respect to the biofilm formation on the fiber core surface was observed. The findings from this study showed that the sensor detection had better sensitivity at near-infrared wavelengths than at visible-light wavelengths. Moreover, the sensitivity of this sensor could be controlled by surface modifications of the core surface through electrostatic interactions, involving a silane coupling layer, polyanions, and polycations. The developed sensor was successfully applied to evaluating of the effectiveness of a commercial biofilm inhibitor.

Fiber Optic Sensor for Real-Time Sensing of Silica Scale Formation in Geothermal Water.

We present a novel fiber optic sensor for real-time sensing of silica scale formation in geothermal water. The sensor is fabricated by removing the cladding of a multimode fiber to expose the core to detect the scale-formation-induced refractive index change. A simple experimental setup was constructed to measure the transmittance response using white light as a source and a spectroscopy detector. A field test was performed on geothermal water containing 980 mg/L dissolved silica at 93 °C in Sumikawa Geothermal Power Plant, Japan. The transmittance response of the fiber sensor decreased due to the formation of silica scale on the fiber core from geothermal water. An application of this sensor in the evaluation of scale inhibitors was demonstrated. In geothermal water containing a pH modifier, the change of transmittance response decreased with pH decrease. The effectiveness of a polyelectrolyte inhibitor in prevention of silica scale formation was easily detectable using the fiber sensor in geothermal water.

A xenogeneic-free system generating functional human gut organoids from pluripotent stem cells.

Functional intestines are composed of cell types from all 3 primary germ layers and are generated through a highly orchestrated and serial developmental process. Directed differentiation of human pluripotent stem cells (hPSCs) has been shown to yield gut-specific cell types; however, these structures do not reproduce critical functional interactions between cell types of different germ layers. Here, we developed a simple protocol for the generation of mature functional intestinal organoids from hPSCs under xenogeneic-free conditions. The stem cell-derived gut organoids produced here were found to contain distinct types of intestinal cells, including enterocytes, goblet cells, Paneth cells, and enteroendocrine cells, that were derived from all 3 germ layers; moreover, they demonstrated intestinal functions, including peptide absorption, and showed innervated bowel movements in response to stimulation with histamine and anticholinergic drugs. Importantly, the gut organoids obtained using this xenogeneic-free system could be stably maintained in culture for prolonged periods and were successfully engrafted in vivo. Our xenogeneic-free approach for generating gut organoids from hPSCs provides a platform for studying human intestinal diseases and for pharmacological testing.

Fundamental study on the development of fiber optic sensor for real-time sensing of CaCO3 scale formation in geothermal water.

This study proposes an optical fiber sensor for calcium carbonate (CaCO3) scale formation in water. The sensor is easily fabricated by removing the cladding of a multimode fiber to expose the core towards the surrounding medium in order to detect refractive index change. A variation of the transmittance response from the high refractive index of CaCO3 which precipitated on the fiber core surface was observed. The proposed setup can be used to analyze the transmittance response over wide range of wavelength using white light as a source and also a spectroscopy detector. The curve of the transmittance percentage over time showed that a fiber core with 200 µm has higher sensitivity as compared to a fiber core with 400 µm. The findings from this study showed that the sensor detection region at near infrared (NIR) wavelengths showed better sensitivity than visible light (VIS) wavelengths. Field tests were conducted using natural geothermal water at Matsushiro, Japan in order to verify the performance of the proposed sensor. The optical response was successfully evaluated and the analytical results confirmed the capability of monitoring scale formation in a geothermal water environment.

Simultaneous multiselective spectroelectrochemical fiber-optic sensor: demonstration of the concept using methylene blue and ferrocyanide.

Herein, we present a novel spectroelectrochemical fiber-optic sensor that combines electrochemistry, spectroscopy, and electrostatic adsorption in three modes of selectivity. The proposed sensor is simple and consists of a gold mesh cover on a multimode fiber optic that uses attenuated total reflection as the optical detection mode. The sensing is based on changes in the attenuation of the light that passes through the fiber-optic core accompanying the electrochemical oxidation-reduction of an analyte at the electrode. Methylene blue and ferrocyanide were used as model analytes to evaluate the performance of the proposed sensor. The optical transmission changes generated by electrochemical manipulation showed a good linear relationship with the concentration and the limits of detection (3σ) for methylene blue and ferrocyanide at 2.0 × 10(-7) and 1.6 × 10(-3) M, respectively. The sensor responses were successfully enhanced with an additional level of selectivity via an electrostatically adsorbed, self-assembled monolayer (SAM), which consisted of a silane coupling layer, a polyanion, and a polycation. The improvement observed in the sensitivity of a SAM-modified fiber-optic sensor was rather encouraging. The optimized sensor had detection limits (3σ) of 8.3 × 10(-9) M for methylene blue and 7.1 × 10(-4) M for ferrocyanide. The developed sensor was successfully applied to the detection of ferrocyanide in simulated nuclear waste.

Molybdenum blue spectrophotometry for trace arsenic in ground water using a soluble membrane filter and calcium carbonate column.

An improved molybdenum blue spectrophotometry using a soluble membrane filter and CaCO(3)-column was proposed for determining arsenic in drinking water supplied from ground water in the presence of phosphate. A 100 mL sample solution containing 0.5 - 10 µg arsenic was passed through a CaCO(3)-column to remove phosphate, arsenate (As(V)). Arsenite (As(III)) which was not retained on the column was oxidized to As(V). As(V) was converted into a heteropolymolybdenum blue anion. The blue anion was collected on a membrane filter as an ion-associate with n-dodecyltrimethylammonium ion by filtration. The filter was dissolved in 2 mL of 2-methoxyethanol. The absorbance of the solution was measured at 810 nm against a reagent blank. Total inorganic arsenic was determined by reducing As(V) to As(III) before the column treatment. The RSDs for 10 µg L(-1) of As(III) and As(V) were 2.9%. Phosphate 0.2 mg L(-1) (as P) and iron 0.1 mg L(-1) did not interfere with the determination of 10 µg L(-1) arsenic. The proposed method was successfully applied to ground waters.

Loss of the Brm-type SWI/SNF chromatin remodeling complex is a strong barrier to the Tat-independent transcriptional elongation of human immunodeficiency virus type 1 transcripts.

To elucidate the epigenetic regulation of Tat-independent human immunodeficiency virus (HIV) transcription following proviral integration, we constructed an HIV type 1 (HIV-1)-based replication-defective viral vector that expresses a reporter green fluorescent protein (GFP) product from its intact long terminal repeat (LTR). We transduced this construct into human tumor cell lines that were either deficient in or competent for the Brm-type SWI/SNF complex. One day after transduction, single cells that expressed GFP were sorted, and the GFP expression profiles originating from each of these clones were analyzed. Unlike clones of the SWI/SNF-competent cell line, which exhibited clear unimodal expression patterns in all cases, many clones originating from Brm-deficient cell lines either showed a broad-range distribution of GFP expression or were fully silenced. The resorting of GFP-negative populations of these isolated clones showed that GFP silencing is either reversible or irreversible depending upon the proviral integration sites. We further observed that even in these silenced clones, proviral gene transcription initiates to accumulate short transcripts of around 60 bases in length, but no elongation occurs. We found that this termination is caused by tightly closed nucleosome-1 (nuc-1) at the 5' LTR. Also, nuc-1 is remodeled by exogenous Brm in some integrants. From these results, we propose that Brm is required for the occasional transcriptional elongation of the HIV-1 provirus in the absence of Tat. Since the Brm-type SWI/SNF complex is expressed at marginal levels in resting CD4+ T cells and is drastically induced upon CD4+ T-cell activation, we speculate that it plays crucial roles in the early Tat-independent phase of HIV transcription in affected patients.

Encouraging effect of cadherin-mediated cell-cell junctions on transfer printing of micropatterned vascular endothelial cells.

We made micropatterned vascular endothelial cells, which have a regular capillary tube-like structure, on a bioactive hydrogel matrix. We applied a stamping method to transfer micropatterned bovine aortic endothelial cells to a growth factor-reduced basement membrane matrix (Matrigel) and type I collagen gel. In this study, we addressed the issues of how to accelerate cell transfer and the effective factors in doing so. We focused on the effects of the cell-substratum and cell-cell adhesiveness prior to applying cultured endothelial cells to a hydrogel matrix on cellular behavior under transfer printing. We found that individual cells cultured sparsely on substrata with different cell adhesivity transferred to Matrigel up to 40%, whereas cells cultured on patterned substrata having lines of 60 mum in width, which involved cell-cell contacts, could transfer homogeneously to Matrigel within a few hours. The morphology of such cells changed from a tape-like monolayer into a thinner, tube-like structure. The speed and the ratio of transfer of micropatterned cells to Matrigel were affected by the period of cell culture on micropatterned substrata. We also found that the intensity of vascular endothelial cadherin staining at cell-cell junctions of micropatterned cells was correlated with cellular behavior when applying them to Matrigel, on which cells formed a tube-like structure or to which cells migrated individually. Furthermore, micropatterned cells made regular tube-like structures when applied to type I collagen gel. Such tube-like endothelial cells had good viability. These findings may be useful for creating in vitro angiogenesis assays and tissue-like constructs that include capillary-like networks of vascular endothelial cells.

Frequent loss of Brm expression in gastric cancer correlates with histologic features and differentiation state.

The mammalian SWI/SNF chromatin remodeling complex, an essential epigenetic regulator, contains either a single Brm or BRG1 molecule as its catalytic subunit. We observed frequent loss of Brm expression but not of BRG1 in human gastric cancer cell lines. Treatment with histone deacetylase inhibitor rescued Brm expression, indicating epigenetic regulation of this gene, and an RNA interference-based colony formation assay revealed antioncogenic properties of Brm. Brm immunostaining of 89 primary gastric cancers showed an obvious reduction in 60 cases (67%) and a severe decrease in 37 cases (42%). Loss of Brm is frequent in the major gastric cancer types (well- or moderately-differentiated tubular adenocarcinoma and poorly-differentiated adenocarcinoma) and positively correlates with the undifferentiated state. Among the minor gastric cancer types, Brm expression persists in signet-ring cell carcinoma and mucinous adenocarcinoma, but a marked decrease is observed in papillary adenocarcinoma. Intestinal metaplasia never shows decreased expression, indicating that Brm is a valid marker of gastric oncogenesis. In contrast, BRG1 is retained in most cases; a concomitant loss of BRG1 and Brm is rare in gastric cancer, contrary to other malignancies. We further show that Brm is required for villin expression, a definitive marker of intestinal metaplasia and differentiation. Via regulating such genes important for gut differentiation, Brm should play significant roles in determining the histologic features of gastric malignancy.