103 research outputs found

    Characterization of common marmoset (Callithrix jacchus) bone marrow-derived mesenchymal stem cells

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    Abstract: Mesenchymal stem cells (MSCs) could be useful for regenerative medicine because they can beharvested easily from the bone marrow of living donors and the cells can be differentiated into adipogenic, osteogenic, and chondrogenic lineages in vitro. To apply MSCs for the medical treatment of human diseases as regenerative medicine, detailed experimental characterization of the cells is required. Recently, a New World primate, the common marmoset (Callithrix jacchus), has been widely used as a new human disease model because of its ease of handling and breeding. Although common marmoset MSCs have been established and will be used in preclinical studies of regenerative medicine, the characteristics of these cells remain unclear. Aiming to characterize common marmoset MSCs further, we harvested common marmoset bone marrow-derived cells (cmBMDCs) from the femurs of newborn males. We revealed that the morphology of the cells was similar to common marmoset fibroblasts, and extracellular matrix components, such as gelatin and fibronectin, were effective for their proliferation and formation of colony-forming unit fibroblasts. Furthermore, we were able to differentiate cmBMDCs into adipocytes, osteocytes, and chondrocytes in vitro, and they expressed the MSCmarkers CD44, CD73, CD90, and CD105, but their expression decreased with increasing passage number. The data demonstrate that cmBMDCs exhibit characteristics of MSCs and thus it would be beneficial to use these cells in preclinical studies

    Impending Gastric Rupture in a Neonate with Gastric Outlet Obstruction due to Malrotation

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    This is a case report of a neonate with impending gastric rupture due to malrotation with gastric outlet obstruction. A preoperative plain abdominal X-ray showed expansion of the gastric bubble. A barium meal demonstrated an unusual bulging of the gastric wall extending from the fundus to the body of the stomach on the greater curvature side and malrotation of the duodenal loop. At operation a malrotation with volvulus and a rupture of the seromuscular layer with bulging of the mucosa in the stomach was found. We think that this unusual bulging of the gastric wall in the barium meal is an important radiological sign of impending gastric rupture

    Correlation Analysis between Antibiotic Resistance Gene Profile and Susceptibility to Gentamicin, Clindamycin, and Minocycline in Clinically Isolated Methicillin-resistant Staphylococcus aureus

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    This study aimed to elucidate retrospectively the correlations between the genome and phenotype in clinical methicillin-resistant Staphylococcus aureus (MRSA) gentamicin (GEN), clindamycin (CLI), and minocycline (MIN) susceptibility using next-generation sequencing (NGS) technology. Ninety two MRSA strains were isolated from individual inpatients treated in Hiroshima University Hospital, Hiroshima, Japan, extracted for their genomic DNA, and sequenced using an Illumina® MiSeq sequencer to obtain their de novo whole-genome assembly. An in silico analysis using ResFinder was performed to obtain the genomic antimicrobial susceptibility profile which was analyzed together with GEN, CLI, and MIN minimum inhibitory concentration (MIC) levels. This study found aac(6’)aph(2”)+, spc+, ermA+, tetM+ MRSA strains were predominant (42/92) and were shown to exhibit >16 mg/L GEN (40/42), >4 mg/L CLI (26/42), and >8 mg/L MIN MIC levels (30/42). Associations between aac(6’)aph(2”) detections and GEN MIC levels (p <0.001), ermA detections and CLI MIC levels (p <0.001), and tetM detections and MIN MIC levels (p <0.001) were revealed in this study. Correlations between simultaneous detections of aac(6’) aph(2”)-spc-ermA-tetM and GEN MIC levels (φc= 0.398, p <0.001), CLI MIC levels (φc= 0.448, p <0.001), and MIN MIC levels (φc= 0.515, p <0.001) were revealed in this study. The genomicphenotypic correlation analyses in this study provided an insight of a rapid antimicrobial detection in MRSA using in silico genomic antimicrobial susceptibility profiling.This research was partially supported by a Grant-in-Aid for Scientific Research (A) (No.15H02567) from the Ministry of Education, Culture, Sports, Science, and Technology and those from Ministry of Health, Labor, and Welfare of the Government of Japan

    Differences in the electric potential of pancreatic head cancer tissues

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    Identifying the electrical properties of cancer relies on the understanding of the electric potential (EP) of cancer tissues. This study aimed to investigate the EP properties in 49 pancreatic head cancer tissues using a digital multimetre. The anode was placed at the central side of the tumour, and the electric potential differences (EPDs) between cancerous and cancerous, cancerous and noncancerous, and noncancerous and noncancerous lesions at approximately 1-cm intervals following resection were evaluated. Pathological evaluation identified 30 of these samples as pancreatic invasive ductal carcinoma (PIDC, 10 without preoperative chemotherapy and 20 after chemotherapy), seven other pancreatic cancers, three tumours of Vater’s ampulla (VA), and eight extrahepatic cholangiocarcinoma (EHCC) samples. We also evaluated the differences in pH for cancerous and noncancerous lesions in nine PIDC samples. Our data suggest that the EP of pancreatic cancerous tissues is higher than that of noncancerous tissues, especially in PIDCs. We also noted that EPD was the highest when comparing cancerous and noncancerous lesions. Additionally, PIDC tissues presented with low pH; the pH difference between cancerous and noncancerous sites was significantly correlated with EPD (P = 0.011). These EPDs were also correlated with tumour size in PIDCs and inversely correlated with their response to chemotherapy. The EP values for both the cancerous and noncancerous sites in both the VA tumours and EHCC samples were not significantly different, whereas EPD in PIDC correlated with tumour extension and viable tumour content, suggesting that EPD might be useful for evaluating the viability and effectiveness of neoadjuvant chemotherapy.This research was partially supported by a Grant-in-Aid for Scientific Research (A) (Nos. 15H02567 and 17H05102) from the Ministry of Education, Culture, Sports, Science, and Technology, and the Ministry of Health, Labour, and Welfare for Japan

    Familial Occurrence of a Congenital Portosystemic Shunt of the Portal Vein

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    A congenital portosystemic shunt of the portal vein is a very rare vascular anomaly associated with the liver. We report the case of a 5-year-old girl with a patent ductus venosus and her 31-year-old mother with a congenital portosystemic shunt. The child presented with a history of an extremely low birth weight in addition to an atrial septal defect and a patent ductus venosus. At the age of 2, she underwent ligation of the ductus venosus. Her mother was also diagnosed with a congenital vascular anomaly at the age of 16. We have followed up and evaluated her asymptomatic mother for 15 years. To our knowledge, this is the first report describing the occurrence of a congenital portosystemic shunt in both a mother and her child

    A robust method for estimating gene expression states using Affymetrix microarray probe level data

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    <p>Abstract</p> <p>Background</p> <p>Microarray technology is a high-throughput method for measuring the expression levels of thousand of genes simultaneously. The observed intensities combine a non-specific binding, which is a major disadvantage with microarray data. The Affymetrix GeneChip assigned a mismatch (MM) probe with the intention of measuring non-specific binding, but various opinions exist regarding usefulness of MM measures. It should be noted that not all observed intensities are associated with expressed genes and many of those are associated with unexpressed genes, of which measured values express mere noise due to non-specific binding, cross-hybridization, or stray signals. The implicit assumption that all genes are expressed leads to poor performance of microarray data analyses. We assume two functional states of a gene - expressed or unexpressed - and propose a robust method to estimate gene expression states using an order relationship between PM and MM measures.</p> <p>Results</p> <p>An indicator 'probability of a gene being expressed' was obtained using the number of probe pairs within a probe set where the PM measure exceeds the MM measure. We examined the validity of the proposed indicator using Human Genome U95 data sets provided by Affymetrix. The usefulness of 'probability of a gene being expressed' is illustrated through an exploration of candidate genes involved in neuroblastoma prognosis. We identified the candidate genes for which expression states differed (un-expressed or expressed) when compared between two outcomes. The validity of this result was subsequently confirmed by quantitative RT-PCR.</p> <p>Conclusion</p> <p>The proposed qualitative evaluation, 'probability of a gene being expressed', is a useful indicator for improving microarray data analysis. It is useful to reduce the number of false discoveries. Expression states - expressed or unexpressed - correspond to the most fundamental gene function 'On' and 'Off', which can lead to biologically meaningful results.</p

    Cell-Free DNA Analysis of Epithelial Growth Factor Receptor Mutations in Lung Adenocarcinoma Patients by Droplet Digital PCR

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    Cell-free DNA (cfDNA) analysis may provide a non-invasive diagnostic approach for lung adenocarcinoma patients. Recently, droplet digital PCR (ddPCR) has been developed as a highly sensitive detection method for a low mutant allele percentage. The ddPCR detection limit for epithelial growth factor receptor (EGFR) mutations was evaluated using cell lines, NCI-H1975 for EGFR L858R point mutation and PC-9 for EGFR E746-A750del. Subsequently, detection of EGFR mutations by ddPCR was performed in tumor DNA (tDNA) and cfDNA samples of 19 lung adenocarcinoma patients whose tumor biopsies were already evaluated for EGFR mutations by clamp PCR (13 of L858R, 3 of E746-750del, and 3 of EGFR negative). In 12 cases, immunohistochemical analysis was performed to quantify the number of EGFR L858R-positive cells rate. EGFR point mutation or deletion were detected in 16 tumor DNA samples. In the measurable cfDNA samples, the rate of detection by ddPCR in cfDNA was 61.5% (8/13) for L858R and 100% (3/3) for E746-A750del. A relative correlation was found between the allele fraction (AF) of tDNA and the number of EGFR L858R-positive cells rate. No correlation was found between the AF of tDNA and AF of cfDNA. In our study, cfDNA mutation detection was not associated with clinicopathological features, but cases with high AF of cfDNA did have metastatic lesions. Our study shows that ddPCR enables cfDNA analysis for EGFR L858R and E746-A750del, with a high detection rate. Therefore, cfDNA analysis using ddPCR may complement to tumor biopsy and is beneficial for precision medicine in lung adenocarcinoma patients

    Fluorescence Image-Guided Navigation Surgery Using Indocyanine Green for Hepatoblastoma

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    In the past decade, navigation surgery using fluorescent indocyanine green (ICG) dye for hepatoblastoma (HB) has been developed for the resection of primary or metastatic tumors. Since HB cells can take up ICG but cannot excrete it to the bile duct, ICG remains in the HB cells, which can be used for navigation by fluorescent activation. The complete resection of the primary tumor as well as metastatic tumors, along with appropriate neoadjuvant and adjuvant chemotherapy, is essential for cure. ICG fluorescence can detect microscopic residual lesions in the primary lesion and identify micro-metastases in the lung or other lesions; consequently, ICG navigation surgery may improve outcomes for patients with HB. The basic technique and recent advances in ICG navigation for HB surgery are reviewed