Generation of a large number of functional dendritic cells from human monocytes expanded by forced expression of cMYC plus BMI1

Anticancer vaccination therapies with monocyte-derived dendritic cells (DC) are widely conducted. A large number of primary monocytes (approximately 10⁸ cells) are needed to generate the number of DC required to achieve an effect upon vaccination, and monocytes are usually purified from peripheral blood mononuclear cells obtained by apheresis procedure, which is somehow invasive for cancer patients. As a means to facilitate the generation of DC for therapeutic use, we herein report a method to amplify human monocytes. We found that lentivirus-mediated transduction of cMYC along with BMI1 induced proliferation of CD14⁺ monocytes derived from 9 out of 12 blood donors, and we named the monocyte-derived proliferating cells CD14-ML. Their proliferation continued for 3–5 weeks in the presence of M-CSF and GM-CSF, resulting in 20–1000-fold amplification. Importantly, the expanded CD14-ML differentiated into fully functional DC (CD14-ML-DC) upon the addition of IL-4 to the culture. We successfully stimulated autologous CD8⁺ T cells with CD14-ML-DC pulsed with cytomegalovirus peptide or MART-1 peptide to generate antigen-specific CTL lines. This is the first report describing the method for in vitro expansion of human peripheral blood monocytes.     

Utility of bedside artificial pancreas for postoperative glycemic control in cardiac surgery

Journal of Artificial Organs - Perioperative hyperglycemia, hypoglycemia, and high glycemic variability are independent risk factors for mortality in critically ill patients. After cardiac surgery,...     

Monte Carlo simulation of the acquisition conditions for 177Lu molecular imaging of hepatic tumors

Annals of Nuclear Medicine - To examine the impact of acquisition time on Lutetium-177 (177Lu) single-photon emission computed tomography (SPECT) images using Monte Carlo simulation. A gamma camera...     

Dab1‐mediated colocalization of multi‐adaptor protein CIN85 with Reelin receptors,ApoER2 and VLDLR,in neurons

Reelin‐Dab1 signaling is indispensable for proper positioning of neurons in mammalian brain. Reelin is a glycoprotein secreted from Cajal‐Reztuis cells in marginal zone of cerebral cortex, and its receptors are Apolipoprotein E receptor 2 (ApoER2) or very low density lipoprotein receptor (VLDLR) expressed on migrating neurons. When Reelin binds to ApoER2 or VLDLR, an adaptor protein Dab1 bound to the receptors undergoes Tyr phosphorylation that is essential for Reelin signaling. We reported previously that Cdk5‐p35 phosphorylates Dab1 at Ser400 and Ser491 and the phosphorylation regulates its binding to CIN85, which is an SH3‐containing multiadaptor protein involved in endocytic downregulation of receptor‐tyrosine kinases. However, the interaction of CIN85 with Dab1 has not been addressed in neurons. We examined here a possibility that CIN85 has a role in Reelin signaling. We found nonpho‐sphorylated Dab1‐mediated colocalization of CIN85 with ApoER2. The colocalization of CIN85 with ApoER2 was increased in neurons stimulated with Reelin repeats 3‐6, an active Reelin fragment. The stimulation recruited CIN85 to domains in plasma membrane where it colocalized with ApoER2 and Dab1 and then to EEA1‐labeled early endosomes in the cytoplasm. In addition, Tyr phosphorylation of Dab1 strengthened the binding to CIN85. These results suggest that CIN85 participates in Reelin signaling through the binding to Dab1.     

Validity and Reproducibility of an Incremental Sit-To-Stand Exercise Test for Evaluating Anaerobic Threshold in Young,Healthy Individuals

Sit-to-stand exercise (STS) is a common activity of daily living. The objectives of the present study were: 1) to assess the validity of aerobic fitness measurements based on anaerobic thresholds (ATs), during incremental sit-to-stand exercise (ISTS) with and without arm support compared with an incremental cycle-ergometer (CE) test; and 2) to examine the reproducibility of the AT measured during the ISTSs. Twenty-six healthy individuals randomly performed the ISTS and CE test. Oxygen uptakes at the AT (AT-VO₂) and heart rate at the AT (AT-HR) were determined during the ISTSs and CE test, and repeated-measures analyses of variance and Tukey’s post-hoc test were used to evaluate the differences between these variables. Pearson correlation coefficients were used to assess the strength of the relationship between AT-VO₂ and AT-HR during the ISTSs and CE test. Data analysis yielded the following correlations: AT-VO₂ during the ISTS with arm support and the CE test, r = 0.77 (p < 0.05); AT-VO₂ during the ISTS without arm support and the CE test, r = 0.70 (p < 0.05); AT-HR during the ISTS with arm support and the CE test, r = 0.80 (p < 0.05); and AT-HR during the ISTS without arm support and the CE test, r = 0.66 (p < 0.05). The AT-VO₂ values during the ISTS with arm support (18.5 ± 1.9 mL·min^-1·kg^-1) and the CE test (18.4 ± 1.8 mL·min^-1·kg^-1) were significantly higher than those during the ISTS without arm support (16.6 ± 1.8 mL·min^-1·kg^-1; p < 0.05). The AT-HR values during the ISTS with arm support (126 ± 10 bpm) and the CE test (126 ± 13 bpm) were significantly higher than those during the ISTS without arm support (119 ± 9 bpm; p < 0.05). The ISTS with arm support may provide a cardiopulmonary function load equivalent to the CE test; therefore, it is a potentially valid test for evaluating AT-VO2 and AT-HR in healthy, young adults.

Key points

• The ISTS is a simple test that varies only according to the frequency of standing up, and requires only a small space and a chair.
• The ISTS with arm support is valid and reproducible, and is a safe test for evaluating AT in healthy young adults.
• For evaluating the AT, the ISTS may serve as a valid alternative to conventional CPX, using either a cycle ergometer or treadmill, in cases where the latter methods are difficult to implement.

Key words: Sit-to-stand, cycle ergometer, anaerobic threshold, correlations, reproducibility, validity     

Hypercholesterolemia as a part of chronic GVHD after allogeneic stem cell transplantation

A 45-year-old female with acute myelogenous leukemia (AML-M6) received an allogeneic stem cell transplantation from an HLA-identical sibling donor in June 2002. Prophylaxis against graft-versus-host disease (GVHD) consisted of cyclosporine (CsA) and short-term methotrexate. Acute GVHD did not occur and CsA was discontinued on day 145 after transplantation. However, soon thereafter she suffered from conjunctivitis, stomatitis and liver dysfunction with hypercholesterolemia and was diagnosed as having chronic GVHD. The liver dysfunction and hypercholesterolemia failed to improve despite the administration of CsA and prednisolone. Atrovastatin was not effective and immunosuppressive therapy for two months including ursodeoxycholic acid finally improved the jaundice and hypercholesterolemia. Although lipid metabolism analysis in this case disclosed the same findings as in other intrahepatic cholestatic liver diseases, the results show that the improvement of hypercholesterolemia in chronic GVHD needs the same treatment as chronic GVHD.     

Idiopathic premature ventricular contractions arising from the pulmonary artery: importance of mapping in the pulmonary artery in left bundle branch block-shaped ventricular arrhythmias.

A patient underwent radiofrequency (RF) catheter ablation of symptomatic idiopathic ventricular contractions (PVCs). RF energy applications at 2 sites in the right ventricular outflow tract (RVOT), where both the earliest ventricular activation and near-perfect pace mapping were obtained, did not abolish the PVC but resulted in changes in the QRS morphology of the PVC. Complete elimination of the PVC was achieved with RF energy application at a site within the pulmonary artery 13 mm above the pulmonary valve, which was greater than 20 mm away from the failed ablation sites within the RVOT.     

Relationships of Fatty Acids,Delta-5 Desaturase Activity,and Lipid Profiles in Men with Acute Coronary Syndrome

Aims: We evaluated the relationship between the ratios of eicosapentaenoic acid and arachidonic acid (EPA/AA), docosahexaenoic acid (DHA)/AA, and delta-5 desaturase activity (D5D) and atherogenic lipid profiles (ALP) and coronary atherosclerosis.Methods: Polyunsaturated fatty acids (PUFA) and ALP were assessed in 436 men with the first episode of acute coronary syndrome (ACS) not take any lipid-lowering drugs. D5D was estimated as the ratio of AA to dihomogamma-linolenic acid (DGLA). These biomarkers were compared between the lower and higher levels of EPA/AA (0.41) or DHA/AA (0.93) according to the levels in Japanese general population. The thrombolysis in myocardial infarction flow (TIMI) grade of the culprit coronary artery was visually estimated during the initial angiography.Results: Approximately 70% of patients had low EPA/AA or DHA/AA. Serum levels of LDL-cholesterol, apolipoprotein B (apoB), and remnant lipoprotein cholesterol (RL-C) were significantly higher in the low EPA/AA or DHA/AA groups, while those of triglycerides and malondialdehyde-modified LDL (MDA-LDL) were significantly higher in the low EPA/AA group alone. The levels of EPA, EPA/AA, DHA/AA, and HbA1c increased and those of DGLA and apoA1 decreased with increasing number of stenotic vessels. Patients with three stenotic coronary vessels or TIMI grade ≥ 1 had significantly higher EPA levels compared with the others. The levels of LDL-cholesterol, non-HDL-cholesterol, triglycerides, small dense LDL-cholesterol, RL-C, MDA-LDL, apoB, and apoE decreased progressively and those of EPA, DHA, EPA/AA and HDL-cholesterol increased as D5D increased.Conclusions: The EPA/AA is a superior risk marker than DHA/AA in term of correlation with ALP in ACS patients.