Morphologic analysis of pulmonary neuroendocrine tumors.

BACKGROUND: Few studies on how to diagnose pulmonary neuroendocrine tumors through morphometric analysis have been reported. In this study, we measured and analyzed the characteristic parameters of pulmonary neuroendocrine tumors using an image analyzer to aid in diagnosis. METHODS: Sixteen cases of typical carcinoid tumor, 5 cases of atypical carcinoid tumor, 15 cases of small cell carcinoma, and 51 cases of large cell neuroendocrine carcinoma were analyzed. Using an image analyzer, we measured the nuclear area, perimeter, and the major and minor axes. RESULTS: The mean nuclear area was 0.318±0.101 µm(2) in typical carcinoid tumors, 0.326±0.119 µm(2) in atypical carcinoid tumors, 0.314±0.107 µm(2) in small cell carcinomas, and 0.446±0.145 µm(2) in large cell neuroendocrine carcinomas. The mean nuclear circumference was 2.268±0.600 µm in typical carcinoid tumors, 2.408±0.680 µm in atypical carcinoid tumors, 2.158±0.438 µm in small cell carcinomas, and 3.247±1.276 µm in large cell neuroendocrine carcinomas. All parameters were useful in distinguishing large cell neuroendocrine carcinoma from other tumors (p=0.001) and in particular, nuclear circumference was the most effective (p=0.001). CONCLUSIONS: Pulmonary neuroendocrine tumors showed nuclear morphology differences by subtype. Therefore, evaluation of quantitative nuclear parameters improves the accuracy and reliability of diagnosis.

Suppression of inflammation by recombinant Salmonella typhimurium harboring CCL22 microRNA.

Atopic dermatitis (AD) is an inflammatory, chronically relapsing, puritic skin disorder. These syndromes result from multifactorial inheritance, with interaction between genetic and environmental factors. In particular, the macrophage-derived chemokine CCL22 is directly implicated in skin inflammatory reactions and its levels are significantly elevated in serum and correlated with disease severity in AD. We tested the suppression of the CCL22 gene by microRNA (miRNA) and observed the effects in mice with inflammation similar to AD. We used Salmonella as a vector to deliver miRNA. The recombinant strain of Salmonella typhimurium expressing CCL22 miRNA (ST-miRCCL22) was prepared for in vivo knockdown of CCL22. ST-miRCCL22 was orally inoculated into mice and the CCL22 gene suppressed with CCL22 miRNA in the activated lymphocytes. IgE and interleukin-4 were inhibited and interferon-γ was induced after treatments with ST-miRCCL22 and CCL22 was suppressed. Further, Th17 cells were suppressed in the atopic mice treated with ST-miRCCL22. These results suggested that suppression of the CCL22 gene using Salmonella induced anti-inflammatory effects.

Therapeutic effects of recombinant Salmonella typhimurium harboring CCL22 miRNA on atopic dermatitis-like skin in mice.

Th-2-biased immune responses are known to play a key role in the pathogenesis of atopic dermatitis. In particular, the macrophage-derived chemokine CCL22 is directly implicated in Th-2-associated skin inflammatory reactions, and its levels are significantly elevated in serum and are correlated with disease severity in atopic dermatitis. In this study, we tested the development of genetic therapeutic options to treat atopic dermatitis using bacteria expressing miRNA. We constructed a recombinant strain of Salmonella typhimurium expressing CCL22 miRNA (ST-miRCCL22) for the in vivo knockdown of CCL22. The CCL22 gene was downregulated with CCL22 miRNA in activated lymphocytes. In mice with a cutaneous disease similar to atopic dermatitis, interleukin-4 was inhibited and interferon-g was induced after treatments with ST-miRCCL22. Furthermore, CCL22 levels were suppressed in the atopic mice treated with ST-miRCCL22. These results suggest that ST-miRCCL22 may be an effective genetic agent for treating atopic dermatitis.

In vitro metabolism of a new neuroprotective agent, KR-31543 in the human liver microsomes: identification of human cytochrome P450.

KR-31543, (2S,3R,4S)-6-amino-4-[N-(4-chlorophenyl)-N-(2-methyl-2H-tetrazol-5-ylmethyl)amino]-3,4-dihydro-2-dimethoxymethyl-3-hydroxy-2-methyl-2H-1-benzopyran, is a new neuroprotective agent for preventing ischemia-reperfusion damage. This study was performed to identify the metabolic pathway of KR-31543 in human liver microsomes and to characterize cytochrome P450 (CYP) enzymes that are involved in the metabolism of KR-31543. Human liver microsomal incubation of KR-31543 in the presence of NADPH resulted in the formation of two metabolites, M1 and M2. M1 was identified as N-(4-chlorophenyl)-N-(2-methyl-2H-tetrazol-5-ylmethyl)amine on the basis of LC/MS/MS analysis with a synthesized authentic standard, and M2 was suggested to be hydroxy-KR-31543. Correlation analysis between the known CYP enzyme activities and the rates of the formation of M1 and M2 in the 12 human liver microsomes have showed significant correlations with testosterone 6beta-hydroxylase activity (a marker of CYP3A4). Ketoconazole, a selective inhibitor of CYP3A4, and anti-CYP3A4 monoclonal antibodies potently inhibited both N-hydrolysis and hydroxylation of KR-31543 in human liver microsomes. These results provide evidence that CYP3A4 is the major isozyme responsible for the metabolism of KR-31543 to M1 and M2.

Surface-bonded fiber optic Sagnac sensors for ultrasound detection.

This paper describes a fiber optic sensor suitable for remote sensing and multi-point detection of ultrasound. This ultrasound sensor is based on the surface-bonded fiber optic Sagnac interferometer with the output fringe visibility of 1; it consists of a laser source, an ordinary single mode fiber delay line, a fiber coupler, a phase modulator and polarization controllers. For the validation of the sensor, surface acoustic waves and Lamb waves are excited by illuminating a steel specimen with an array of Q-switched Nd:YAG laser-generated line sources and the measurement of laser-generated ultrasonic waves are performed on the specimen surface using the surface-mounting fiber optic Sagnac sensor. The surface-bonded fiber optic sensor developed in this study has a simple configuration for detection of ultrasonic waves. Effectiveness of surface-bonded fiber optic Sagnac sensors for remote sensing of ultrasound and in situ monitoring of structures is investigated. The capability of multi-point detection of ultrasound by this Sagnac sensor is also discussed.

Simultaneous determination of sildenafil and its active metabolite UK-103,320 in human plasma using liquid chromatography-tandem mass spectrometry.

A liquid chromatography-tandem mass spectrometric method for the simultaneous determination of sildenafil and its active N-demethylated metabolite, UK-103,320 in human plasma was developed. Sildenafil, UK-103,320 and the internal standard (DA-8159) were extracted from human plasma with dichloromethane at basic pH. A reverse-phase LC separation was performed on Luna phenylhexyl column with the mixture of acetonitrile-ammonium formate (10 mM, pH 6.0) (60:40, v/v) as mobile phase. The detection of analytes was performed using an electrospray ionization tandem mass spectrometry in the multiple reaction-monitoring mode. The lower limits of quantification for sildenafil and UK-103,320 were 2.0 ng/ml. The method showed a satisfactory sensitivity, precision, accuracy, recovery and selectivity.

Semi-micro high-performance liquid chromatographic analysis of tiropramide in human plasma using column-switching.

A rapid and sensitive column-switching semi-micro high-performance liquid chromatography method was developed for the direct analysis of tiropramide in human plasma. The plasma sample (100 microl) was directly injected onto Capcell Pak MF Ph-1 precolumn where deproteinization and analyte fractionation occurred. Tiropramide was then eluted into an enrichment column (Capcell Pak UG C(18)) using acetonitrile-potassium phosphate (pH 7.0, 50 mM) (12:88, v/v) and was analyzed on a semi-micro C(18) analytical column using acetonitrile-potassium phosphate (pH 7.0, 10 mM) (50:50, v/v). The method showed excellent sensitivity (limit of quantification 5 ng/ml), and good precision (C.V.

Metabolism of a new neuroprotective agent for ischemia-reperfusion damage, KR-31543 in the rat using liquid chromatography/electrospray mass spectrometry.

KR-31543, (2S,3R,4S)-6-amino-4-[N-(4-chlorophenyl)-N-(2-methyl-2H-tetrazol-5-ylmethyl)amino]-3,4-dihydro-2-dimethoxymethyl-3-hydroxy-2-methyl-2H-1-benzopyran is a new neuroprotective agent for ischemia-reperfusion damage. The in vitro and in vivo metabolism of KR-31543 in rats has been studied by LC-electrospray mass spectrometry. Rat liver microsomal incubation of KR-31543 in the presence of NADPH resulted in the formation of a metabolite M1. M1 was identified as N-(4-chlorophenyl)-N-(2-methyl-2H-tetrazol-5-ylmethyl)amine on the basis of LC-MS/MS analysis with the synthesized authentic standard. Rat CYP3A1 and 3A2 are the major CYP isozymes involved in the formation of M1.

Non-contact detection of laser-generated surface acoustic waves using fiber optic Sagnac interferometer.

A high powered Q-switched Nd:YAG laser was used to excite the surface waves, and an optical fiber sensor was used to detect the out-of-plane displacements due to the propagating waves. This sensor is based on the fiber optic Sagnac interferometer, which has the path-matched configuration and does not require active stabilization. Quadrature phase bias between two interfering laser beams in the Sagnac loop is applied by controlling the birefringence in an optical path using a fiber polarization controller. A stable quadrature phase bias can be confirmed by observing the interferometer output according to the change of phase bias. Additional signal processing is not needed for the detection of ultrasonic waves using the Sagnac interferometer. The performance of the fiber optic Sagnac interferometer was investigated, and laser-generated surface wave signals were detected using this fiber optic sensor. The developed fiber optic sensor configured in this study is very simple and is effective for non-contact detection of ultrasonic waves.

Noncontact detection of ultrasonic waves using fiber optic Sagnac interferometer.

This paper describes a fiber optic sensor suitable for noncontact detection of ultrasonic waves. This sensor is based on the fiber optic Sagnac interferometer, which has a path-matched configuration and does not require active stabilization. Quadrature phase bias between two interfering laser beams in the Sagnac loop is applied by controlling the birefringence using a fiber polarization controller. A stable quadrature phase bias can be confirmed by observing the interferometer output according to the change of phase bias. Additional signal processing is not needed for the detection of ultrasonic waves using the Sagnac interferometer. Ultrasonic oscillations produced by conventional ultrasonic piezoelectric transducers were successfully detected, and the performance of this interferometer was investigated by a power spectrum analysis of the output signal. Based on the validation of the fiber optic Sagnac interferometer, noncontact detection of laser-generated surface waves was performed. The configured Sagnac interferometer is very effective for the detection of small displacement with high frequency, such as ultrasonic waves used in conventional nondestructive testing (NDT).