The Development of a New Setup for Video-assisted Thoracic Surgery

In order to accomplish video-assisted thoracic surgery (VATS) in a much easier and safer way, especially for assistant operators, we have developed a new display system for VATS. The original thoracoscope has been designed for this new system. The monitor is fixed at approximately 10 cm away from the surface of the chest wall just above the operative field. In using this procedure, the operator and assistants can see the patient and the monitor at the same time. According to this new idea, the previous problem in the area of hand–eye coordination and the three-dimensional understanding of this procedure can be improved compared to the image of the conventional thoracoscopy, because it is not necessary for the operator and assistants to look up at the monitors. When the thoracoscopy was placed in an adequate position to resect the target pathology, this new system led to good and easy handling of instruments, as it was with the standard thoracotomy

The role of infection in the development of non-valvular atrial fibrillation: Up-regulation of Toll-like receptor 2 expression levels on monocytes

SummaryMany studies have suggested that inflammation may participate in the pathogenesis of non-valvular atrial fibrillation (AF). However, it has been unknown by exposure to what the inflammation is caused. Recently, we reported that Toll-like receptor 2 (TLR2) level on monocytes was significantly up-regulated in viral and bacterial infections, but not in non-infectious inflammatory states. Our purpose was to test the hypothesis that expression of TLR2 levels may be up-regulated in patients with non-valvular AF. A total of 48 consecutive patients with non-valvular AF who were hospitalized for catheter ablation were enrolled in this study. TLR2 levels were assayed by using flow-cytometric analysis and compared with volunteers in sinus rhythm (control group, n=24). Additionally, C-reactive protein (CRP) and interleukin-6 (IL-6) levels were assayed, and the left atrial volume indexes (LAVI) in the non-valvular AF group were measured. The results demonstrated that TLR2 levels in the non-valvular AF group were significantly higher than in the control group (median, 4682 vs. 3866 sites/cell; P<0.01). Moreover, non-valvular AF patients had significantly higher IL-6 levels than controls. However, there was no significant difference in CRP levels between the two groups. It was observed in 44 AF patients, in whom pulmonary vein isolation was confirmed to be successful, that the LAVI significantly diminished 1 month after ablation (median, 33.6 vs. 29.5ml/m2; P<0.001), but not the TLR2 and IL-6 levels. Our results implied that an infectious inflammation may participate in the pathogenesis of non-valvular AF

Circulating CD3+HLA-DR+ Extracellular Vesicles as a Marker for Th1/Tc1-Type Immune Responses

Extracellular vesicles (EVs) are known to contain unique proteins that reflect the cells of origins. Activated T cells are reported to secrete various EVs. To establish T cell subset-specific biomarkers, we performed proteomic analysis with Th1- and Th2-derived EVs and identified HLA-DR as a Th1-dominated EV membrane protein. We designed a measurement system for CD3+CD4+, CD3+CD8+, and CD3+HLA-DR+ EVs to specifically determine EV subpopulations derived from CD4+, CD8+, and Th1-type T cells, respectively. In vitro analysis showed that CD3+CD4+ EVs and CD3+CD8+ EVs were selectively secreted from activated CD4+ and CD8+ T cells, respectively, and that CD3+HLA-DR+ EVs were actively secreted from not only Th1 but also activated CD8+ T (probably mostly Tc1) cells. To evaluate the clinical usefulness of these EVs, we measured the serum levels in patients with inflammatory diseases, including Epstein-Barr virus (EBV, n=13) infection, atopic dermatitis (AD, n=10), rheumatoid arthritis (RA, n=20), and osteoarthritis (OA, n=20) and compared the levels with those of healthy adults (n=20). CD3+CD4+ EVs were significantly higher in all of EBV infection, AD, RA, and OA while CD3+CD8+ EVs were higher in EBV infection, lower in RA, and not different in AD and OA relative to the control. The levels of CD3+HLA-DR+ EVs were markedly higher in EBV infection and significantly lower in AD. The results suggest that these EV subpopulations reflect in vivo activation status of total CD4+, total CD8+, and Th1/Tc1-type T cells, respectively, and may be helpful in T cell-related clinical settings, such as cancer immunotherapy and treatment of chronic infection, autoimmune diseases, and graft-versus-host disease

Mast cells derived from human induced pluripotent stem cells are useful for allergen tests

Background: Several methods have been developed to detect allergen-specific IgE in sera. The passive IgE sensitization assay using human IgE receptor-expressing rat cell line RBL-2H3 is a powerful tool to detect biologically active allergen-specific IgE in serum samples. However, one disadvantage is that RBL-2H3 cells are vulnerable to high concentrations of human sera. Only a few human cultured cell lines are easily applicable to the passive IgE sensitization assay. However, the use of human induced pluripotent stem cells (iPSCs) to generate human mast cells (MCs) has not yet been reported. Methods: The nuclear factor-kappa B (NF-κB)-responsive luciferase reporter gene was stably introduced into a human iPSC line 201B7, and the transfectants were induced to differentiate into MCs (iPSC-MCs). The iPSC-MCs were sensitized overnight with sera from subjects who were allergic to cedar pollen, ragweed pollen, mites, or house dust, and then stimulated with an extract of corresponding allergens. Activation of iPSC-MCs was evaluated by β-hexosaminidase release, histamine release, or luciferase intensity. Results: iPSCs-MCs stably expressed high-affinity IgE receptor and functionally responded to various allergens when sensitized with human sera from relevant allergic subjects. This passive IgE sensitization system, which we termed the induced mast cell activation test (iMAT), worked well even with undiluted human sera. Conclusions: iMAT may serve as a novel determining system for IgE/allergens in the clinical and research settings. Keywords: Allergen-specific IgE, Allergy test, Human mast cells, Induced pluripotent stem cells, Luciferas

Non-IgE,-IgG4 Antibody to Japanese Cedar Pollen Allergens:Comparison of Its Prevalence and Titers between Pollinosis Patients and Non-Patients

Background: IgG antibody to allergens in the serum of pollinosis patients is not routinely measured, because there are no simple methods for assaying small amounts of specific IgG in the serum ; so far only IgE and IgG4 antibodies have been assayed. In this study, we used a reverse-sandwich ELISA for measuring specific non-IgE,-IgG4 (probably, mainly IgG1) to Japanese cedar pollen allergens, and compared the antibody-positive rates and geometric mean antibody titers between pollinosis patients and non-patients (healthy individuals). Methods: Antibodies to two major allergens of Japanese cedar, Cry j 1 and Cry j 2, were assayed by the following methods : specific non-IgE,-IgG4 was measured by a reverse-sandwich ELISA ; specific IgE and IgG4 were measured by indirect ELISAs. Results: We detected specific non-IgE,-IgG4 in both the patients and non-patients. In comparison to the IgE antibody, which was detected in a small proportion of the non-patients with low titers, the non-IgE,-IgG4 antibody was present in a higher proportion with higher titers among both the patients and non-patients. Conclusions: Healthy individuals had specific non-IgE,-IgG4 to Japanese cedar allergens in a higher proportion than the specific IgE. The non-IgE,-IgG4 antibody assay may be useful in studies on the prevalence of allergen-specific antibody responders and may help in clarifying the natural history of pollinosis