Methods CD4+CD3+ and CD8+CD60+CD45RO±CD45RA±IL-4±IFNγ± T cells in blood of serum IgE+ (fluoroimmunoassay) ragweed sensitized (RS) and IgE- nonallergic humans expressing receptors for CSP C3a and C5a ...(CD88) (n=2-3/group) were determined by flow cytometry. Conclusions The presence of receptors for C3a on CD4+ and CD8+CD60+CD45RO+ IL-4+ T cells required for induction of ragweed specific memory IgE responses suggests that C3a may play an important role in induction of these responses.
Cells were collected by vigorous pipetting and analyzed by flow cytometry (Fortessa) for surface CD33 expression (anti-CD33 PE, BD Biosciences) and intracellular iNOS expression (rabbit monoclonal ...anti-human iNOS) (abcam), followed by goat anti-rabbit IgG (Alexa Fluor 488)(abcam). Incubation for 18 hrs with CYT+D3 significantly increased CD33+iNOS+ monocytes in PBMC from IgE+ adults (27.5 vs 15.65%; p<0.016), but not iNOS expression in PBMC from IgE- subjects (6.5 vs 8.7%; p>0.05) (Mann-Whitney U-test) Conclusions Measurement of intracellular iNOS by flow cytometry in CD33+ monocytes detects increased expression in the presence of cytokines and vitamin D3 in IgE+ adults.
While BMI does not correlate with FeNO, a biomarker of allergic asthma, it is not known how BMI relates to blood monocyte nitric oxide responses in adults ± allergy/asthma. Cells were collected by ...vigorous pipetting and analyzed for surface CD33 expression (anti-CD33 PE, BD Biosciences) and intracellular iNOS expression (rabbit monoclonal anti-human iNOS) (Abcam), followed by goat anti-rabbit IgG (Alexa Fluor 488)(abcam) using flow cytometry (LSR Fortessa).
Intravascular large B-cell lymphoma (IVLBCL) is a distinct disease entity with the peculiar characteristic that tumor cells proliferate within vessels. Despite recent advances in understanding the ...disease from clinical aspects, the underlying pathogenesis remains unknown. Here we demonstrate analyses of IVLBCL biology using four xenograft mouse models established from primary IVLBCL samples. In all four models, the main characteristic of IVLBCL tumor cell proliferation within vessels was retained. Time-lapse engraftment analyses revealed that the tumor cells initially engrafted and proliferated in the sinusoids and vessels in the liver and then engrafted and proliferated in multiple organs. Intriguingly, serial passage of tumor cells from the adrenal gland of a transplanted mouse developed from primary patient bone marrow cells into a second mouse showed that the tumor cells mainly distributed into the adrenal gland in the second mouse, implying the existence of clonal selection and/or evolution at engraftment of a specific organ. Gene expression profiling analyses demonstrated that the gene set associated with cell migration was enriched for normal peripheral blood B cells, indicating that inhibition of cell migration might be involved in IVLBCL pathogenesis. In conclusion, the mouse xenograft models described here are essential tools for uncovering IVLBCL biology.
Autonomous underwater vehicles (AUVs) are frequently used for deep-water ocean applications such as surveying and cable laying, where accurate control of vehicle depth and attitude is needed. The ...water level in the onboard ballast tanks is typically manually set for neutral buoyancy before each mission, while the vehicle is on the surface. The ballast tank contents are not normally adjusted to control vehicle depth and orientation while the AUV is in operation. As a result, vehicle trajectory and orientation is exclusively controlled using the vehicle's control surfaces during a mission. The challenges with controlling the depth and trim of an underwater vehicle include nonlinear hydrodynamic forces, as well as inherent time delays (latencies) associated with water tank level changes and valve adjustments. Furthermore, small changes in the location of the vehicle's center of gravity (i.e., due to the deployment of the AUV's payload equipment) can reduce the control authority of the AUV's control surfaces. To meet these challenges, this paper proposes two unique variable ballast system (VBS) control approaches. The first proposed VBS controller changes the weight of the AUV to help control vehicle depth and vertical (inertial) velocity. The second proposed VBS controller attempts to shift the center of gravity x G along the body-fixed x -(longitudinal)-axis to reduce depth and pitch angle error while restoring control authority to the bowplane and sternplane deflection fins. The ballasting system consists of two water tanks positioned aft and forward of amidships. The ballast tanks are then automatically filled or emptied of ocean water as desired. Numerical simulations have been carried out on a 2-D underwater vehicle simulator to test and compare the performance of the proposed ballast and fin deflection control systems. The simulation results show that, for the assumptions and conditions tested, the proposed controllers are capable of achieving a setpoint depth and pitch angle with minimal error by effectively utilizing the ballast tanks and deflection fins. As a result, the work presented in this paper helps increase the autonomy of large AUVs on long-duration missions.
Methods We determined the numbers of (l) CD4+IL-4+ and CD8+CD60±IL-4+ T cells and subP receptor+ (NK-1R) leukocyte subsets in blood of ragweed sensitized humans (n=6); (2) CD45+, CD4+CD3+, CD8+CD60±, ...CD19+, CD16/56+, and CD14+ cells expressing p38MAPK in PBMC ± 15-30 min incubation with PMA ± subP; and (3) the effect of subP on specific memory IgE responses by PBMC cultured for 0-12 days ± ragweed antigen ± subP ± antibodies to subP receptors (flow cytometry; ELISA) (data expressed as % T cell subsets, % total lymphocytes or monocytes, ng/ml) Results SubP did not affect CD4+IL-4+ or CD8+CD60+IL-4+ T cells but suppressed CD45+p38MAPK+ lymphocytes (70%) (including CD4+, CD8+, CD19+, CD16/56+ p38MAPK+ cells), and specific memory IgE responses, with antibodies to subP receptors reversing this effect.
Methods Blood/serum was obtained from ragweed sensitized humans (n=6) and CD4+ and CD8+CD60+ T cell expression of p38MAPK (MESF), CD4+IL-4+ and CD8+CD60+IL-4+ T cell numbers and IgE levels (IU/ml) ...were determined (flow cytometery, fluoroenzymeimmunoassay).
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