Long-chain saturated fatty acids in breast milk are associated with the pathogenesis of atopic dermatitis via induction of inflammatory ILC3s.

Breastfeeding influences the immune system development in infants and may even affect various immunological responses later in life. Breast milk provides a rich source of early nutrition for infant growth and development. However, the presence of certain compounds in breast milk, related to an unhealthy lifestyle or the diet of lactating mothers, may negatively impact infants.

Chromosomal translocation t(11;14) and p53 deletion induced by the CRISPR/Cas9 system in normal B cell-derived iPS cells.

Multiple myeloma (MM) cells are derived from mature B cells based on immunoglobulin heavy chain (IgH) gene analysis. The onset of MM is often caused by a reciprocal chromosomal translocation (cTr) between chr 14 with IgH and chr 11 with CCND1. We propose that mature B cells gain potential to transform by reprograming, and then chromosomal aberrations cause the development of abnormal B cells as a myeloma-initiating cell during B cell redifferentiation.

Investigation of the cumulative number of chromosome aberrations induced by three consecutive CT examinations in eight patients.

In our previous study, we found that chromosomes were damaged by the radiation exposure from a single computed tomography (CT) examination, based on an increased number of dicentric chromosomes (Dics) formed in peripheral blood lymphocytes after a CT examination. We then investigated whether a cumulative increase in the frequency of Dics and chromosome translocations (Trs) formation could be observed during three consecutive CT examinations performed over the course of 3-4 years, using lymphocytes in peripheral bloods of eight patients (five males and three females; age range 27-77 years; mean age, 64 years). The effective radiation dose per CT examination estimated from the computational dosimetry system was 22.

Induction of t(11;14) IgH enhancer/promoter- gene translocation using CRISPR/Cas9.

Chromosomal translocation is a key process in the oncogenic transformation of somatic cells. Previously, artificial induction of chromosomal translocation was performed using homologous recombination-mediated loxP labeling of target regions followed by Cre-mediated recombination. Recent progress in genome editing techniques has facilitated the easier induction of artificial translocation by cutting two targeted genome sequences from different chromosomes.

Dose-response curves for analyzing of dicentric chromosomes and chromosome translocations following doses of 1000 mGy or less, based on irradiated peripheral blood samples from five healthy individuals.

In terms of biological dosimetry at the time of radiation exposure, the dicentric chromosome (Dic) assay (DCA) is the gold standard for assessing for the acute phase and chromosome translocation (Tr) analysis is the gold standard for assessing the chronic phase. It is desirable to have individual dose-response curves (DRCs) for each laboratory because the analysis criteria differ between laboratories. We constructed the DRCs for radiation dose estimation (with three methods) using peripheral blood (PB) samples from five healthy individuals.

Establishment of induced pluripotent stem cells from normal B cells and inducing AID expression in their differentiation into hematopoietic progenitor cells.

B cell derived induced pluripotent stem cells (BiPSCs) were recently established from peripheral blood B cells by the simultaneous transfection of Yamanaka factors (Oct3/4, Sox2, Klf4, c-Myc) and C/EBPα using a Sendai virus vector. Here, using a different method, we established BiPSCs with immunoglobulin heavy chain (IgH) gene rearrangement from normal B cells purified from lymph nodes. The critical points of our method are pre-stimulation of B cells with IL-21 and CD40-ligand (CD40L), followed by consecutive transfection of highly concentrated Yamanaka factors using a retroviral vector.

Academic Responses to Fukushima Disaster.

Since radiation accidents, particularly nuclear disasters, are rarer than other types of disasters, a comprehensive radiation disaster medical curriculum for them is currently unavailable. The Fukushima compound disaster has urged the establishment of a new medical curriculum in preparation for any future complex disaster. The medical education will aim to aid decision making on various health risks for workers, vulnerable people, and residents addressing each phase in the disaster.

The great importance of normalization of LC-MS data for highly-accurate non-targeted metabolomics.

The non-targeted metabolomics analysis of biological samples is very important to understand biological functions and diseases. LC combined with electrospray ionization-based MS has been a powerful tool and widely used for metabolomic analyses. However, the ionization efficiency of electrospray ionization fluctuates for various unexpected reasons such as matrix effects and intraday variations of the instrument performances.

Analysis of chromosome translocation frequency after a single CT scan in adults.

We recently reported an increase in dicentric chromosome (DIC) formation after a single computed tomography (CT) scan (5.78-60.27 mSv: mean 24.

Increase in dicentric chromosome formation after a single CT scan in adults.

Excess risk of leukemia and brain tumors after CT scans in children has been reported. We performed dicentric chromosome assay (DCAs) before and after CT scan to assess effects of low-dose ionizing radiation on chromosomes. Peripheral blood (PB) lymphocytes were collected from 10 patients before and after a CT scan.

Direct metabolomics for plant cells by live single-cell mass spectrometry.

Live single-cell mass spectrometry (live MS) provides a mass spectrum that shows thousands of metabolite peaks from a single live plant cell within minutes. By using an optical microscope, a cell is chosen for analysis and a metal-coated nanospray microcapillary tip is used to remove the cell's contents. After adding a microliter of ionization solvent to the opposite end of the tip, the trapped contents are directly fed into the mass spectrometer by applying a high voltage between the tip and the inlet port of the spectrometer to induce nanospray ionization.

Identification of low-dose responsive metabolites in X-irradiated human B lymphoblastoid cells and fibroblasts.

Ionizing radiation (IR) induces cellular stress responses, such as signal transduction, gene expression, protein modification, and metabolite change that affect cellular behavior. We analyzed X-irradiated human Epstein-Barr virus-transformed B lymphoblastoid cells and normal fibroblasts to search for metabolites that would be suitable IR-responsive markers by Liquid Chromotography-Mass spectrometry (LC-MS). Mass spectra, as analyzed with principal component analysis, showed that the proportion of peaks with IR-induced change was relatively small compared with the influence of culture time.

Metabolomic analysis of dynamic response and drug resistance of gastric cancer cells to 5-fluorouracil.

Metabolomics has developed as an important new tool in cancer research. It is expected to lead to the discovery of biomarker candidates for cancer diagnosis and treatment. The current study aimed to perform a comprehensive metabolomic analysis of the intracellular dynamic responses of human gastric cancer cells to 5-fluorouracil (5-FU), referencing the mechanisms of drug action and drug resistance.

Molecular and functional analysis of cellular phenomena using single-cell mass spectrometry.

Single-cell analysis has attracted attention in many fields of biological studies as a tool to survey the precise mechanisms of cellular and molecular behavior. The development of sensitive mass spectrometry allows the study of molecules in single cells or small regions. Matrix-assisted laser desorption/ionization imaging mass spectrometry and secondary-ion mass spectrometry use in situ ionization of specimens on sample plates to visualize molecular distributions as images from mass spectra.

Drug metabolite heterogeneity in cultured single cells profiled by pico-trapping direct mass spectrometry.

Aim: We investigated the heterogeneity of tafluprost metabolism in primary human hepatocytes at a single-cell level by live single-cell mass spectrometry (MS).

Materials & Methods: Picoliter volumes of cytoplasm were analyzed by nano-electrospray ionization MS in order to obtain single-cell metabolite profiles. The subcellular components of a single tafluprost-treated human hepatocyte were isolated and the single-cell metabolite profile was compared with those of traditional bulk hepatocyte analysis.

Direct drug metabolism monitoring in a live single hepatic cell by video mass spectrometry.

The metabolism of anti-breast cancer drug, tamoxifen, in a single human hepatocellular carcinoma cell, HepG2, was directly monitored by a video-mass spectroscope. The cytoplasm, a vacuole or nucleus of the cell was directly sucked by a nano-spray tip under a video-microscope, and then was introduced into a mass spectrometer. Unchanged drug molecules were found in cytoplasm and a vacuole, but the metabolites were only found in the cytoplasm.

In situ molecular analysis of plant tissues by live single-cell mass spectrometry.

We report the development of a rapid, direct molecular analysis of live, single plant cells viewed under a video microscope in their natural environment. A nanoelectrospray tip was used to extract the contents of a single leaf, stem, or petal cell from Pelargonium zonale, and the samples were analyzed on an Orbitrap mass spectrometer by nanoelectrospray ionization. Around a thousand m/z peaks belonging to metabolites and other compounds in each sample were obtained and processed by using statistical tools to find the cell specific molecular peaks.

Mass spectrometry for cellular and tissue analyses in a very small region.

The analysis of biological specimens is now expected to shift from multi-cell analysis to single cells to know about the molecular and cellular behavior precisely. Mass spectrometric approaches to clarify molecular profiles in a cell or very small region have been developed. Such techniques include matrix-assisted laser desorption ionization imaging mass spectrometry (MALDI-IMS), secondary-ion mass spectrometry (SIMS), single-cell MALDI and live single-cell mass spectrometry.

A novel signaling pathway associated with Lyn, PI 3-kinase and Akt supports the proliferation of myeloma cells.

Interleukin-6 (IL-6) is a growth factor for human myeloma cells. We have recently found that in myeloma cells the activation of both signal transducer and activator of transcription (STAT) 3 and extracellular signal-regulated kinase (ERK) 1/2 is not sufficient for the IL-6-induced proliferation, which further requires the activation of the src family kinases, such as Lyn. Here we showed that the Lyn-overexpressed myeloma cell lines had the higher proliferative rate with IL-6 and the enhanced activation of the phosphatidylinositol (PI) 3-kinase and Akt.

Direct single-cell molecular analysis of plant tissues by video mass spectrometry.

Direct and quick molecular analysis of live plant single cells viewed under a video-microscope has been developed in their intact undamaged environment using nano-electrospray tip and mass spectrometers. This method has been successful in identifying specific molecules in plant live single-cell analysis by mass spectrometry, and opens the possibility to compare different cell types from different tissues with morphological evidence in undamaged plants.

Live single-cell metabolomics of tryptophan and histidine metabolites in a rat basophil leukemia cell.

A direct and rapid metabolic analysis of a live single cell was performed by live single-cell video-mass spectrometry. The contents of the cytoplasm and a granule were sucked into a nano-electrospray ionization (nano-ESI) tip, and were directly introduced into a Q-TOF mass spectrometer by nano-spray after the addition of an ionization solvent. The metabolic pathways and the locations of tryptophan and histidine metabolites were traced by this method for a cultured rat basophil leukemia cell line (RBL-2H3).

Ectopic cyclin D1 overexpression increases chemosensitivity but not cell proliferation in multiple myeloma.

We established a myeloma cell line (RPMI8226) with cyclin D1 overexpression in which the transfected cyclin D1 gene was stably expressed. D1 transfectants showed down-regulation of cyclin D2. Cell proliferation analysis did not show any differences among RPMI8226, mock control, and D1 transfectants.