545 research outputs found

    Non-cross-linking gold nanoparticle aggregation as a detection method for single-base substitutions

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    Aggregation of DNA-modified gold nanoparticles in a non-cross-linking configuration has extraordinary selectivity against terminal mismatch of the surface-bound duplex. In this paper, we demonstrate the utility of this selectivity for detection of single-base substitutions. The samples were prepared through standard protocols: DNA extraction, PCR amplification and single-base primer extension. Oligonucleotide-modified nanoparticles correctly responded to the unpurified products from the primer extension: aggregation for the full match and dispersion for all the mismatches. Applicability of this method to genomic DNA was tested with five human tumor cell lines, and verified by conventional technologies: mass spectrometry and direct sequencing. Unlike the existing methods for single-base substitution analysis, this method does not need specialized equipments, and opens up a new possibility of point-of-care diagnosis for single-nucleotide polymorphisms

    Selectively Expanded of CD4+ T Cells from Peripheral Blood Mononuclear Cells Induced by Interleukin-2 Plus Anti-CD3 : Effects on the Cytotoxicity, the Proliferation and the Modulation of HLA Antigens and Intercellular Adhesion Molecule-1 against Human Malignant Melanoma Cell Lines

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    We selected human CD4? T cells using immunomagnetic beads from periph-eral blood mononuclear cells of 5 healthy volunteers, and cultured with recom-binant interleukin-2 (RIL-2) plus anti-CD3 culture supernatants. After success-ful generation of the CD4? T cell cultures, we examined the therapeutic value for human malignant melanoma. Through flow cytometry analysis, ??Cr-release assay and proliferation assay, we found that a) the total increase in 7 to 10 day cultures ranged from 30 to 100-fold ; b) the expanded cells were mostly CD2?, CD3?, CD4?, CD8?, CD25?, CD29?, CD56? and T cell receptor (α/β)? T cell ; c) some of them possessed moderate levels of natural killer cell and lymphokine -activated killer cell activities ; d) culture supernatants significantly inhibited the growth of cultured human melanoma cell lines ; e) culture supernatants enhanced surface expression of class I, class II and intercellular adhesion molecule-1 on cultured human melanoma cell lines ; and f) culture supernatants contained large amounts of cytokines such as IL-2, IL-6 and interferon-γ. These results sug-gested that selectively expanded CD4+ T cells may be available as a new strategy in the future human adoptive immunotherapy

    Is Adenosine Deaminase in Pleural Fluid a Useful Marker for Differentiating Tuberculosis from Lung Cancer or Mesothelioma in Japan, a Country with Intermediate Incidence of Tuberculosis?

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    The objective of this study was to evaluate the utility of the determination of adenosine deaminase (ADA) level in pleural fluid for the differential diagnosis between tuberculous pleural effusion (TPE) and malignant pleural effusion (MPE) in Japan, a country with intermediate incidence of tuberculosis (TB). We retrospectively reviewed the clinical records of 435 patients with pleural effusion and investigated their pleural ADA levels as determined by an auto analyzer. ROC analysis was also performed. The study included patients with MPE (n=188), TPE (n=124), benign nontuberculous pleural effusion (n=94), and pleural effusion of unknown etiology (n=29). The median ADA level in the TPE group was 70.8U/L, which was significantly higher than that in any other groups (p<0.05). The area under the curve (AUC) in ROC analysis was 0.895. With a cut-off level for ADA of 36U/L, the sensitivity, specificity, positive predictive value, and negative predictive value were 85.5%, 86.5%, 69.7%, and 93.6%, respectively. As many as 9% of patients with lung cancer and 15% of those with mesothelioma were false-positive with this ADA cutoff setting. Although the ADA activity in pleural fluid can help in the diagnosis of TPE, it should be noted that some cases of lung cancer or mesothelioma show high ADA activity in geographical regions with intermediate incidence of TB, in contrast to high prevalence areas

    Axillary artery injury combined with delayed brachial plexus palsy due to compressive hematoma in a young patient: a case report

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    <p>Abstract</p> <p>Introduction</p> <p>Axillary artery injury in the shoulder region following blunt trauma without association with either shoulder dislocation or fracture of the humeral neck has been previously reported. Axillary artery injury might also be accompanied with brachial plexus injury. However, delayed onset of brachial plexus palsy caused by a compressive hematoma associated with axillary injury after blunt trauma in the shoulder region has been rarely reported. In previous reports, this condition only occurred in old patients with sclerotic vessels. We present a case of a young patient who suffered axillary artery injury associated with brachial plexus palsy that occurred tardily due to compressive hematoma after blunt trauma in the shoulder region without association of either shoulder dislocation or humeral neck fracture.</p> <p>Case presentation</p> <p>A 16-year-old male injured his right shoulder in a motorbike accident. On initial physical evaluation, the pulses on the radial and ulnar arteries in the affected arm were palpable. Paralysis developed later from 2 days after the injury. Functions in the right arm became significantly impaired. Angiography showed complete occlusion of the axillary artery. Magnetic resonance imaging demonstrated a mass measuring 4 × 5 cm that was suspected to be a hematoma compressing the brachial plexus in a space between the subscapular muscle and the pectoralis minor muscle. Surgery was performed on the third day after injury. In intraoperative observations, the axillary artery was occluded with thrombus along 5 cm; a subscapular artery was ruptured; the brachial plexus was compressed by the hematoma. After evacuation of the hematoma, neurolysis of the brachial plexus, and revascularization of the axillary artery, the patient had an excellent functional recovery of the affected upper limb, postoperatively.</p> <p>Conclusion</p> <p>Surgeons should be aware that axillary artery injuries may even occur in young people after severe blunt trauma in the shoulder region and can be associated with secondary brachial plexus injury due to a hematoma. For treatment in cases with progressive nervous deficit after trauma, not only reconstruction of the injured artery but also immediate evacuation of the hematoma, and exploration of the brachial plexus are necessary to avoid irreversible neurological damage.</p

    Regulated growth of diatom cells on self-assembled monolayers

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    We succeeded in regulating the growth of diatom cells on chemically modified glass surfaces. Glass surfaces were functionalized with -CF(3), -CH(3), -COOH, and -NH(2 )groups using the technique of self-assembled monolayers (SAM), and diatom cells were subsequently cultured on these surfaces. When the samples were rinsed after the adhesion of the diatom cells on the modified surfaces, the diatoms formed two dimensional arrays; this was not possible without the rinsing treatment. Furthermore, we examined the number of cells that grew and their motility by time-lapse imaging in order to clarify the interaction between the cells and SAMs. We hope that our results will be a basis for developing biodevices using living photosynthetic diatom cells

    A Case of Cystic Basal Cell Carcinoma Which Shows a Homogenous Blue/Black Area under Dermatoscopy

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    Basal cell carcinoma (BCC) is the most common skin tumor and contains several different histopathological types. Here, we report a case of cystic basal cell carcinoma, which is relatively rare and might be clinically misdiagnosed. A dermatoscopic examination of the case revealed a homogenous blue/black area usually not seen in BCC. We reviewed 102 BCC cases resected and diagnosed at Sapporo Medical University Hospital between April 2005 and March 2010. Among them, only three were the cystic type
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