Evaluating the Non-Invasive Measurement of Apple Aroma Using Electronic Nose Device through Comparison with Direct Mass Spectrometry, Sugar Content, and Ripeness Measurements.

To compare apple aroma intensities, apples were analyzed from the calyx side (on the opposite side of the stem) using an electronic nose (e-nose) sensor device and direct mass spectrometry. The results indicated that the sensor value tended to increase in accordance with the total intensity of apple aroma components measured by direct mass spectrometry. In addition, the e-nose sensor values for apple aroma did not correlate with the sugar content and ripeness measurements using optical sensors. Moreover, the relative standard deviations of repeatability and intermediate precision in the measurement of apple flavor (apple lip balm) were within 1.36-9.96%. Similar to the utilization of sugar content and ripeness values, the aroma measured from the calyx side can be potentially used for apple evaluation.

Comparison of Cheese Aroma Intensity Measured Using an Electronic Nose (E-Nose) Non-Destructively with the Aroma Intensity Scores of a Sensory Evaluation: A Pilot Study.

Cheese aroma is known to affect consumer preference. One of the methods to measure cheese aroma is the use of an electronic nose (e-nose), which has been used to classify cheese types, production areas, and cheese ages. However, few studies have directly compared the aroma intensity scores derived from sensory evaluations with the values of metal oxide semiconductor sensors that can easily measure the aroma intensity. This pilot study aimed to investigate the relationship between sensory evaluation scores and e-nose values with respect to cheese aroma. Five types of processed cheese (two types of normal processed cheese, one type containing aged cheese, and two types containing blue cheese), and one type of natural cheese were used as samples. The sensor values obtained using the electronic nose, which measured sample aroma non-destructively, and five sensory evaluation scores related to aroma (aroma intensity before intake, during mastication, and after swallowing; taste intensity during mastication; and remaining flavor after swallowing (lasting flavor)) determined by six panelists, were compared. The e-nose values of many of the tested cheese types were significantly different, whereas the sensory scores of the one or two types of processed cheese containing blue cheese and those of the natural cheese were significantly different. Significant correlations were observed between the means of e-nose values and the medians of aroma intensity scores derived from the sensory evaluation testing before intake, during mastication, and after swallowing. In particular, the aroma intensity score during mastication was found to have a linear relationship with the e-nose values (Pearson's R = 0.983). In conclusion, the e-nose values correlated with the sensory scores with respect to cheese aroma intensity and could be helpful in predicting them.

Real-time breath ammonia measurement using a novel cuprous bromide sensor device in patients with chronic liver disease: a feasibility and pilot study.

We developed a small portable sensor device using a p-type semiconductor cuprous bromide (CuBr) thin film to measure breath ammonia in real time with highsensitivity and selectivity. Breath ammonia is reportedly associated with chronic liver disease (CLD). We aimed to assess the practical utility of the novel CuBr sensor device for exhaled breath ammonia and the correlation between breath and blood ammonia in CLD patients. This was a feasibility and pilot clinical study of 21 CLD patients and 18 healthy volunteers. Breath ammonia was directly and quickly measured using the novel CuBr sensor device and compared with blood ammonia measured at the same time. CLD patients had significantly higher breath ammonia levels than healthy subjects (p = 1.51 × 10-3), with the level of significance being similar to that for blood ammonia levels (p= 0.024). Significant differences were found in breath and blood ammonia between the healthy and cirrhosis groups (p = 2.97 × 10-3 and 3.76 × 10-3, respectively). Significant, positive correlations between breath and blood ammonia were noted in the CLD group (R = 0.747, p = 1.00 × 10-4), healthy/CLD group (R = 0.741, p = 6.75 × 10-8), and cirrhosis group (R = 0.744, p = 9.52 × 10-4). In conclusion, the newly developed, easy-to-use, and small portable CuBr sensor device was able to non-invasively measure breath ammonia in real time. Breath ammonia measured using the device was correlated with blood ammonia and the presence of liver cirrhosis, and might be an alternative surrogate biomarker to blood ammonia.

Temozolomide does not influence the transcription or activity of matrix metalloproteinases 9 and 2 in glioma cell lines.

Glioblastoma multiforme (GBM) is a treatment-resistant malignancy with poor prognosis. Temozolomide (TMZ) is widely used as a first-line drug for GBM. Although this improves patient prognosis, it does not completely eradicate the tumour. Even after total surgical resection, GBM can exhibit uncontrollable invasiveness at the tumour margins owing to activation of matrix metalloproteinases (MMPs) such as MMP-2 and -9; these degrade collagen IV in the basement membrane, which normally prevents cancer invasion. TMZ induces DNA damage and activates transcription factors including c-jun, c-fos, nuclear factor-κß, and early growth response protein-1, which have putative binding sites on the MMP-9 promoter. TMZ may therefore enhance tumour invasion by stimulating MMP-9 transcription and enzymatic activity. To test this hypothesis, we investigated MMP-2 and -9 mRNA transcription and activity in GBM cell lines treated with TMZ. Human A172 GBM cells were exposed to TMZ (25% and 50% inhibitory concentrations) for 24 or 48h; cell cycle distribution and mRNA levels of MMP-2 and -9 were evaluated using flow cytometry and semi-quantitative reverse transcription PCR, respectively. MMP-2 and -9 enzymatic activities were assessed using gelatin zymography in human A172 and U373 MG GBM cells exposed to TMZ under the same conditions. TMZ altered A172 cell cycle distribution, but not MMP-2 or -9 mRNA levels. TMZ did not affect MMP-2 or -9 enzymatic activities in A172 or U373 MG cells. These findings indicated that TMZ is therefore unlikely to promote GBM invasiveness.

The Powdering Process with a Set of Ceramic Mills for Green Tea Promoted Catechin Extraction and the ROS Inhibition Effect.

For serving green tea, there are two prominent methods: steeping the leaf or the powdered leaf (matcha style) in hot water. The purpose of the present study was to reveal chemical and functional differences before and after the powdering process of green tea leaf, since powdered green tea may contribute to expanding the functionality because of the different ingesting style. In this study, we revealed that the powdering process with a ceramic mill and stirring in hot water increased the average extracted concentration of epigallocatechin gallate (EGCG) by more than three times compared with that in leaf tea using high-performance liquid chromatography (HPLC) and liquid chromatography-tandem mass Spectrometry (LC-MS/MS) analyses. Moreover, powdered green tea has a higher inhibition effect of reactive oxygen species (ROS) production in vitro compared with the same amount of leaf tea. Our data suggest that powdered green tea might have a different function from leaf tea due to the higher catechin contents and particles.

Cell Membrane Perforation with Photosensitizer and a Brush-shaped Soft-polymer Sheet Using a Malignant Glioma Cell Line.

BACKGROUND: Transduction of foreign molecules into cells is an important technique to investigate the functions of corresponding molecules and/or targets. Recently, a mass-producible nanoprinting perforator was devised enabling for large-scale, high-performance drug or nucleic-acid transfer into cells without cell damage. Since little is known on the performance of the system, we investigated its effects on a malignant glioma cell line. MATERIALS AND METHODS: Photosensitization was performed by the Cell Stamper CP-01. The malignant U373MG glioma cell line was used for transduction. RESULTS: Photosensitization transduced FITC-conjugated albumin into cells. Trypan blue inclusion test demonstrated membrane disintegration by the procedure and scanning electron microscopy disclosed perforation of the cell membrane. CONCLUSION: Local oxidation reaction during the nanoprinting caused reversible membrane perforation. Morphological findings from the current study support the above mechanism, therefore the specific printing system might be convenient for transduction of foreign molecules into malignant glioma cells.

Improving the performance of an electronic nose by wine aroma training to distinguish between drip coffee and canned coffee.

Coffee aroma, with more than 600 components, is considered as one of the most complex food aromas. Although electronic noses have been successfully used for objective analysis and differentiation of total coffee aromas, it is difficult to use them to describe the specific features of coffee aroma (i.e., the type of smell). This is because data obtained by electronic noses are generally based on electrical resistance/current and samples are distinguished by principal component analysis. In this paper, we present an electronic nose that is capable of learning the wine related aromas using the aroma kit "Le Nez du Vin," and the potential to describe coffee aroma in a similar manner comparable to how wine experts describe wine aroma. The results of our investigation showed that the aromas of three drip coffees were more similar to those of pine and honey in the aroma kit than to the aromas of three canned coffees. Conversely, the aromas of canned coffees were more similar to the kit coffee aroma. In addition, the aromatic patterns of coffees were different from those of green tea and red wine. Although further study is required to fit the data to human olfaction, the presented method and the use of vocabularies in aroma kits promise to enhance objective discrimination and description of aromas by electronic noses.

Abnormal number cell division of human thyroid anaplastic carcinoma cell line, SW 1736.

Cell division, during which a mother cell usually divides into two daughter cells during one cell cycle, is the most important physiological event of cell biology. We observed one-to-four cell division during imaging of live SW1736 human thyroid anaplastic carcinoma cells transfected with a plasmid expressing the hybrid protein of green fluorescent protein and histone 2B (plasmid eGFP-H2B). Analysis of the images revealed a mother cell divided into four daughter cells. And one of the abnormally divided daughter cells subsequently formed a dinucleate cell.

Secretion of urocortin I by human glioblastoma cell lines, possibly via the constitutive pathway.

Corticotropin-releasing factor (CRF) and its family of peptides, i.e., urocortins (UCNs), play a critical role in systemic and peripheral stress-response systems and are widely expressed not only in normal tissues but also in various types of cancer cells. Given limited understanding of the mechanism of UCN I secretion, we investigated the UCN I secretory pathway in human neural stem cells (HNSCs) and in two glioblastoma cell lines, e.g., A172 and U-138 MG. Immunoreactivities for CRF receptors were detected in A172 glioblastoma cells, but not in HNSCs or U-138 glioblastoma cells, while UCN I immunoreactivity was detected in A172 and U-138 MG glioblastoma cell lines by both light field and electron microscopy. Interestingly, electron microscopy revealed UCN I immunoreactivtiy in vesicle-like structures in the plasma membrane of the glioblastoma cells. Tracking of a hybrid fluorescent protein containing a UCN I signal peptide expressed in A172 human glioblastoma cells revealed that fluorescence in secretory granules could be decreased by cycloheximide (100µg/ml), indicating that the forward transport of secretory granules containing fluorescent protein was not altered by the inhibition of protein synthesis by cycloheximide. Retrograde transport and the fusion of fluorescent granules in A172 human glioblastoma cells was induced by brefeldin A (10µg/ml), indicating that UCN I secretory granules may be transported via the constitutive pathway. Based on these results, it appears that UCN I is secreted from human glioblastoma cells by exocytosis through constitutive secretory granules, indicating that transcription of UCN I mRNA may be correlated to secretion of UCN I protein.

Effects of silica and titanium oxide particles on a human neural stem cell line: morphology, mitochondrial activity, and gene expression of differentiation markers.

Several in vivo studies suggest that nanoparticles (smaller than 100 nm) have the ability to reach the brain tissue. Moreover, some nanoparticles can penetrate into the brains of murine fetuses through the placenta by intravenous administration to pregnant mice. However, it is not clear whether the penetrated nanoparticles affect neurogenesis or brain function. To evaluate its effects on neural stem cells, we assayed a human neural stem cell (hNSCs) line exposed in vitro to three types of silica particles (30 nm, 70 nm, and <44 µm) and two types of titanium oxide particles (80 nm and < 44 µm). Our results show that hNSCs aggregated and exhibited abnormal morphology when exposed to the particles at concentrations = 0.1 mg/mL for 7 days. Moreover, all the particles affected the gene expression of Nestin (stem cell marker) and neurofilament heavy polypeptide (NF-H, neuron marker) at 0.1 mg/mL. In contrast, only 30-nm silica particles at 1.0 mg/mL significantly reduced mitochondrial activity. Notably, 30-nm silica particles exhibited acute membrane permeability at concentrations =62.5 µg/mL in 24 h. Although these concentrations are higher than the expected concentrations of nanoparticles in the brain from in vivo experiments in a short period, these thresholds may indicate the potential toxicity of accumulated particles for long-term usage or continuous exposure.

Cholangiocarcinoma cell line TK may be useful for the pharmacokinetic study of the chemotherapeutic agent gemcitabine.

Cholangiocarcinoma is a disease with a poor prognosis. A human cholangiocarcinoma cell line, TK, was previously established to enable further understanding of the disease. We conducted this investigation to determine whether or not the TK line is useful for pharmacokinetic study of the chemotherapeutic agent gemcitabine (GEM). Along with the BXPC3 human pancreatic adenocarcinoma cell line, the sensitivity to and effects on the TK cell line of GEM were compared. The influence of deoxycytidine kinase (dCK) transduction was also comparatively investigated. The effects of GEM in terms of drug sensitivity of the TK cell line, cell cycle and levels of transcripts of key enzymes were comparable to the BXPC3 cell line. Responses to the drug were similar in both cell lines. In contrast to pancreatic carcinoma, cell lines for research on cholangiocarcinoma have been limited. This study suggests the application of the TK cell line to the pharmacokinetic study of the chemosensitization of therapeutic drugs, such as GEM.

An Extracellular Matrix Molecule, Secreted by the Epithelial-Mesenchymal Transition is Associated With Lymph Node Metastasis of Thyroid Papillary Carcinoma.

BACKGROUND: Papillary thyroid carcinoma often has lymph node metastasis, compared with follicular thyroid carcinoma. The study showed that epithelial-mesenchymal transition occurs in carcinoma cells during the first stage of metastasis, where some extracellular matrix molecules are secreted in large quantities. Sialic acid carried by fibronectin as the antigen of the monoclonal antibody (MoAb) JT-95, was detected in 90% of papillary thyroid carcinoma cases, and in a few follicular thyroid carcinomas, in the extracellular matrix of thyroid carcinoma cells. OBJECTIVES: The current study was conducted to investigate the association between increasing the number of extracellular matrix molecules, fibronectin, and lymph node metastasis. We also co-cultured a thyroid carcinoma cell line and lymphocyte cell line, with and without MoAb JT-95, in order to investigate the mechanism of cell to cell interaction. PATIENTS AND METHODS: Immunostaining with JT-95 was performed in 45 papillary thyroid carcinoma cases, and 20 follicular type tumors, to investigate the association between the quantity of fibronectin expression and the frequency of lymph node metastasis. The thyroid carcinoma cell line (SW1736), which secreted fibronectin, and the B cell-lymphoma cell line (Daudi), which held integrin on the cell surface, were co-cultured to observe the adhesion of cells to each other. The SW1736 cell line, pretreated with JT-95, was also co-cultured with the Daudi cell line. RESULTS: There were 39 cases with lymph node metastasis in 59 malignant tumors, and 0 cases in 6 benign follicular type tumors. The staining scores by JT-95 of the 39 tumors with lymph node metastasis were 5+ in eight cases and 6+ in 31 cases. On the other hand, the scores of 20 malignant tumors without lymph node metastasis were < 4+ in all of the cases. In the co-cultured assay, numerous adhesions were observed between the SW1736 and Daudi cells. In contrast, the inhibition of adherences was observed in proportion to the concentrations of JT-95. CONCLUSIONS: Increased fibronectin expression in thyroid malignancies is correlated with lymph node metastasis.

Morphological study of the TK cholangiocarcinoma cell line with three-dimensional cell culture.

Cholangiocarcinoma is an intractable carcinoma originating from the bile duct epithelium. To gain an understanding of the cell biology of cholangiocarcinoma, in vitro cell culture is valuable. However, well­characterized cell lines are limited. In the present study, the morphology of the TK cholangiocarcinoma cell line was analyzed by three­dimensional culture. Dispersed TK cells were injected into a gelatin mesh scaffold and cultivated for 3­20 days. The morphology of the TK cells was investigated by phase­contrast microscopy, optical microscopy, scanning electron microscopy (SEM) and transmission electron microscopy (TEM). TK cells were observed to proliferate three-dimensionally in the scaffold. The cells exhibited a globoid structure and attached to the scaffold. The SEM observation demonstrated typical microvilli and plicae on the surface of the structure. Light microscopy and TEM confirmed intercellular and cell­to­scaffold attachment in the three­dimensional mesh. The culture also exhibited the formation of a duct-like structure covered by structured microvilli. In conclusion, three­dimensional culture of TK cells demonstrated the morphological characteristics of cholangiocarcinoma in vitro. Production of high levels of carbohydrate antigen (CA)19­9, CA50 and carcinoembryonic antigen was previously confirmed in the TK cell line. As a characteristic morphology was demonstrated in the present study, the TK cholangiocarcinoma cell line may be useful as an experimental model for further study of cholangiocarcinoma.

Cell-based in vitro blood-brain barrier model can rapidly evaluate nanoparticles' brain permeability in association with particle size and surface modification.

The possibility of nanoparticle (NP) uptake to the human central nervous system is a major concern. Recent reports showed that in animal models, nanoparticles (NPs) passed through the blood-brain barrier (BBB). For the safe use of NPs, it is imperative to evaluate the permeability of NPs through the BBB. Here we used a commercially available in vitro BBB model to evaluate the permeability of NPs for a rapid, easy and reproducible assay. The model is reconstructed by culturing both primary rat brain endothelial cells and pericytes to support the tight junctions of endothelial cells. We used the permeability coefficient (P(app)) to determine the permeability of NPs. The size dependency results, using fluorescent silica NPs (30, 100, and 400 nm), revealed that the Papp for the 30 nm NPs was higher than those of the larger silica. The surface charge dependency results using Qdots® (amino-, carboxyl-, and PEGylated-Qdots), showed that more amino-Qdots passed through the model than the other Qdots. Usage of serum-containing buffer in the model resulted in an overall reduction of permeability. In conclusion, although additional developments are desired to elucidate the NPs transportation, we showed that the BBB model could be useful as a tool to test the permeability of nanoparticles.

Expression of mRNAs of Urocortin in the STKM-1 gastric cancer cell line.

BACKGROUND: Urocortin is analogous to corticotrophin-releasing factors (CRFs) and a member of the CRF family. We previously demonstrated that urocortin mRNAs were expressed in both human and rat glioma cell lines, and that some of these lines transcribed the receptors. We hypothesize that urocortin might also be expressed in a gastric cancer cell line. The aim of the present study was to clarify the expression of mRNAs of urocortin1 (UCN1), -2 and -3 and of CRF and CRF receptors 1 and 2 in a gastric cancer cell line. MATERIALS AND METHODS: STKM-1 a poorly-differentiated adenocarcinoma cell line was used. Transcripts in the cells were analyzed using cDNA. The fluctuation of mRNA with cellular stress, such as the one caused by a chemotherapeutic agent, serum supplementation and forskolin was examined. RESULTS: Transcripts of UCN1, -2 and CRFR2 were expressed. No changes in transcription of UCN1 and UCN2 were observed with cellular stress. However, expression of CRFR2 mRNA transcripts significantly increased after an initial 24-h exposure to forskolin. CONCLUSION: Expression of the mRNAs of UCN1, 2 and CRFR2 was confirmed in the human gastric cancer cell line, STKM-1. Although the quantity of CRFR2 transcripts varied with forskolin, the overall transcription pattern was not influenced by cellular stimuli.

Discrimination method of the volatiles from fresh mushrooms by an electronic nose using a trapping system and statistical standardization to reduce sensor value variation.

Electronic noses have the benefit of obtaining smell information in a simple and objective manner, therefore, many applications have been developed for broad analysis areas such as food, drinks, cosmetics, medicine, and agriculture. However, measurement values from electronic noses have a tendency to vary under humidity or alcohol exposure conditions, since several types of sensors in the devices are affected by such variables. Consequently, we show three techniques for reducing the variation of sensor values: (1) using a trapping system to reduce the infering components; (2) performing statistical standardization (calculation of z-score); and (3) selecting suitable sensors. With these techniques, we discriminated the volatiles of four types of fresh mushrooms: golden needle (Flammulina velutipes), white mushroom (Agaricus bisporus), shiitake (Lentinus edodes), and eryngii (Pleurotus eryngii) among six fresh mushrooms (hen of the woods (Grifola frondosa), shimeji (Hypsizygus marmoreus) plus the above mushrooms). Additionally, we succeeded in discrimination of white mushroom, only comparing with artificial mushroom flavors, such as champignon flavor and truffle flavor. In conclusion, our techniques will expand the options to reduce variations in sensor values.

Evaluation of anti-inflammatory drug-conjugated silicon quantum dots: their cytotoxicity and biological effect.

Silicon quantum dots (Si-QDs) have great potential for biomedical applications, including their use as biological fluorescent markers and carriers for drug delivery systems. Biologically inert Si-QDs are less toxic than conventional cadmium-based QDs, and can modify the surface of the Si-QD with covalent bond. We synthesized water-soluble alminoprofen-conjugated Si-QDs (Ap-Si). Alminoprofen is a non-steroid anti-inflammatory drug (NSAID) used as an analgesic for rheumatism. Our results showed that the "silicon drug" is less toxic than the control Si-QD and the original drug. These phenomena indicate that the condensed surface integration of ligand/receptor-type drugs might reduce the adverse interaction between the cells and drug molecules. In addition, the medicinal effect of the Si-QDs (i.e., the inhibition of COX-2 enzyme) was maintained compared to that of the original drug. The same drug effect is related to the integration ratio of original drugs, which might control the binding interaction between COX-2 and the silicon drug. We conclude that drug conjugation with biocompatible Si-QDs is a potential method for functional pharmaceutical drug development.

Detection of Aeromonas hydrophila in liquid media by volatile production similarity patterns, using a FF-2A electronic nose.

A technique for rapid detection of pathogenic microorganisms is essential for the diagnosis of associated infections and for food safety analysis. Aeromonas hydrophila is one such food contaminant. Several methods for rapid detection of this pathogen have been developed; these include multiplex polymerase chain reaction assays and the colony overlay procedure for peptidases. However, these conventional methods can only be used to detect the microorganisms at high accuracy after symptomatic onset of the disease. Therefore, in the future, simple pre-screening methods may be useful for preventing food poisoning and disease. In this paper, we present a novel system for the rapid detection of the microorganism A. hydrophila in cultured media (in <2 h), with the use of an electronic nose (FF-2A). With this electronic nose, we detected the changes of volatile patterns produced by A. hydrophila after 30 min culture. Our calculations revealed that the increased volatiles were similar to the odours of organic acids and esters. In future, distinctive volatile production patterns of microorganisms identified with the electronic nose may have the potential in microorganism detection.

Expression of mRNAs of urocortin and corticotropin-releasing factor receptors in malignant glioma cell lines.

BACKGROUND: Urocortin and corticotropin-releasing factors (CRFs) and their receptors are expressed in many organs, including the central nervous system. In this study, the expression of mRNAs of urocortin 1, 2, 3, and CRF and CRF receptors 1 and 2 in malignant glioma, was examined. MATERIALS AND METHODS: The RNAs of human and rat glioma cell lines were isolated. Transcripts in these cells were analyzed using cDNA. In addition, the effects of proliferative and cytotoxic stimulation by serum supplementation, ionizing radiation, and the antineoplastic agent temozolomide were investigated. RESULTS: Human and rat cells transcribed urocortin. CRF receptors were detected in human glioma cells. When human KNS42 cells were exposed to stimulation, transcription was altered according to the specific condition. CONCLUSION: Expression of mRNAs of urocortin and CRF receptors was confirmed in human glioma cell lines. Although the quantities of transcripts varied with the proliferative and cytotoxic stimulation, the overall transcription pattern was not influenced by these stimuli.

Suppression of aldosterone synthesis and secretion by ca(2+) channel antagonists.

Aldosterone, a specific mineralocorticoid receptor (MR) agonist and a key player in the development of hypertension, is synthesized as a final product of renin-angiotensin-aldosterone system. Hypertension can be generally treated by negating the effects of angiotensin II through the use of angiotensin-converting enzyme inhibitors (ACE-Is) or angiotensin II type 1 receptor antagonists (ARBs). However, the efficacy of angiotensin II blockade by such drugs is sometimes diminished by the so-called "aldosterone breakthrough" effect, by which ACE-Is or ARBs (renin-angiotensin system (RAS) inhibitors) gradually lose their effectiveness against hypertension due to the overproduction of aldosterone, known as primary aldosteronism. Although MR antagonists are used to antagonize the effects of aldosterone, these drugs may, however, give rise to life-threatening adverse actions, such as hyperkalemia, particularly when used in conjunction with RAS inhibitors. Recently, several groups have reported that some dihydropyridine Ca(2+) channel blockers (CCBs) have inhibitory actions on aldosterone production in in vitro and in the clinical setting. Therefore, the use of such dihydropyridine CCBs to treat aldosterone-related hypertension may prove beneficial to circumvent such therapeutic problems. In this paper, we discuss the mechanism of action of CCBs on aldosterone production and clinical perspectives for CCB use to inhibit MR activity in hypertensive patients.