Deep vein thrombosis after major orthopedic surgery in Taiwan: A prospective cross-sectional study and literature review.

Background/purpose: Postoperative venous thromboembolism is an important complication in Taiwan. We prospectively investigated the occurrence of deep vein thrombosis (DVT) after major orthopedic surgery without pharmacologic thromboprophylaxis in a cohort of 120 patients (46 males, 74 females, median age 71 years) at our institute.

Methods: Color duplex compression ultrasonography (CUS) was used to detect DVT before and after the operation, while contrast venography was performed postoperatively for comparison and validation.

Modulation of the anticancer activities of paclitaxel by Cremophor micelles.

The objective of this study was to determine the effect of Cremophor (CrEL) on the antineoplastic effect induced by paclitaxel (PTX). Fluorescence spectroscopy, employing pyrene as a probe, was used to determine the critical micelle concentration (CMC) of CrEL. EL4 murine thymoma cells and MDA-MB-231 human breast cancer cells were treated with PTX in different concentrations of CrEL.

Recruitment of bone marrow CD11bGr-1 cells by polymeric nanoparticles for antigen cross-presentation.

The objective of this study was to investigate the function of poly(lactic-co-glycolic acid) (PLGA) nanoparticles (NPs) on the activation of antigen-specific CD8 T cell responses via the CD11bGr1 myeloid subpopulations in murine bone marrow (BM). PLGA NPs containing ovalbumin (OVA) were fabricated by the double-emulsion method. The CD11bGr-1Ly-6C and CD11bGr-1Ly-6C subsets from mice bone marrow were sorted and treated with the PLGA/OVA NPs, followed by co-culture with the carboxyfluorescein succinimidyl ester (CFSE)-labelled OT-I CD8 cells.

Cellular biodistribution of polymeric nanoparticles in the immune system.

The biodistribution of polymeric nanoparticles (NPs) is of crucial importance in the development of nanoparticle-based vaccine delivery or immunotherapy for cancer. The purpose of this study was to investigate the kinetics of cellular biodistribution of polymeric NPs in the immune system. Polystyrene (PS) yellow-green nanoparticles (YG-NPs) 500nm in diameter were intravenously (i.

Activation of Antigen-Specific CD8(+) T Cells by Poly-DL-Lactide/Glycolide (PLGA) Nanoparticle-Primed Gr-1(high) Cells.

Purpose: The aim of this study was to investigate the induction of antigen-specific T cell activation and cell cycle modulation by a poly-DL-lactide/glycolide (PLGA) nanoparticle (NP)-primed CD11b(+)Gr-1(high) subset isolated from mouse bone marrow.

Methods: PLGA NPs containing the ovalbumin (OVA) antigen were prepared using the double emulsion and solvent evaporation method, and protein release rate and cell viability were determined. The Lin2(¯)CD11b(+)Gr-1(high)Ly6c(low) (Gr-1(high)) subset was sorted from the bone marrow of C57BL/6 J mice by fluorescence-activated cell sorting (FACS) and co-cultured with OT-I CD8(+) splenic T cells.

The role of tomatine adjuvant in antigen delivery for cross- presentation.

The aim of this study was to investigate the use of tomatine adjuvant to deliver soluble antigen for crosspresentation by bone marrow-derived dendritic cells (BMDCs). BMDCs were incubated with tomatine adjuvantovalbumin (OVA) complex and analyzed for antigen uptake by flow cytometry. Adjuvant-induced cell death was examined in situ by terminal deoxynucleotidyl transferase dUTP nick end labelling (TUNEL) assay.

Encapsulation of poly(D,L-lactide) microparticles with polyelectrolyte multilayers for antigen delivery.

Poly(D,L-lactide) (PLA) microparticles containing the ovalbumin (OVA) model antigen were prepared by the double-emulsion and solvent evaporation method, followed by encapsulation with alternating layers of the polyelectrolytes, consisting of protamine sulfate and dextran sulfate of various molecular weights. The physicochemical properties, including particle size and zeta potentials, were characterised. Treatment of mouse macrophages with surface-modified PLA microparticles stimulated the generation of reactive oxygen species (ROS), such as hydrogen peroxide and superoxide, which was detected by the fluorescent probes, 2',7'-dichlorofluorescein diacetate (DCFH-DA) and hydroethidine (HE).

Antigen delivery for cross priming via the emulsion vaccine adjuvants.

The function of emulsion adjuvants in vaccine antigen delivery remains unclear. To investigate the roles of emulsion adjuvants in cross presentation of exogenous antigens, a series of emulsions were prepared for both in vitro and in vivo studies. Bone marrow-derived dendritic cells (BMDCs) were treated with the adjuvants and analyzed by flow cytometry for the expression of costimulatory molecules.

Delivery of DNA-based cancer vaccine with polyethylenimine.

DNA-based vaccine directed to tumor-specific antigens is an attractive strategy in cancer prevention and therapy. In view of the poor immunogenicity of most tumor-associated antigens, studies were carried out here to examine the adjuvant effect of polyethylenimine (PEI), a cationic polymer widely used in non-viral gene delivery, on the efficacy of cancer vaccine strategy. Treatment of animals with the DNA/PEI complexes resulted in antigen-specific cell lysis and activation of B3Z cells, an ovalbumin/K(b)-specific cytotoxic clone that recognizes the target cells through the class I major histocompatibility complex (MHC) molecules.

The effect of poly(D,L-lactide-co-glycolide) microparticles with polyelectrolyte self-assembled multilayer surfaces on the cross-presentation of exogenous antigens.

A surface-engineered particulate delivery system for exogenous antigens was developed in this study. Poly(d,l-lactide-co-glycolide) (PLGA) microparticles containing ovalbumin (OVA) or fluorescein isothiocyanate-conjugated bovine serum albumin (FITC-BSA) were fabricated by the double emulsion and solvent evaporation method. Encapsulation of the PLGA microparticles was performed by physisorption of multilayers of oppositely charged polyelectrolytes, including polyethylenimine (PEI) and dextran sulfate.

Glucose- and metabolically regulated hepatic insulin gene therapy for diabetes.

Purpose: The purpose of this study was to examine glucose- and metabolically modulation of insulin secretion by rAAV-mediated gene delivery in vitro and in vivo.

Materials And Methods: A recombinant adeno-associated virus vector (rAAV) containing a furin-mutated human insulin gene, driven by the rat insulin I promoter, was used in this study. Glucose-responsive secretion of human insulin was determined by treating rAAV-transduced Huh7 human hepatoma cells with varying concentrations of glucose, with or without insulin secretagogues.

Enhanced antigen delivery via cell death induced by the vaccine adjuvants.

The objective of this study was to examine the effect of cell death induced by the emulsion adjuvants on the in vitro delivery of antigens into the antigen-presenting cells (APCs). J774A.1 murine macrophage-like cells, serving as the APCs, were pulsed with various vaccine adjuvants, and incubated with fluorescein isothiocyanate-conjugated bovine serum albumin (BSA-FITC), with or without adjuvants-pretreated EL4 murine thymoma cells, followed by analysis by flow cytometry and fluorescence microscopy.

Effect of polyethylenimine on recombinant adeno-associated virus mediated insulin gene therapy.

Background: Recombinant adeno-associated virus (rAAV) is becoming a promising vector for gene therapy for type I diabetes. The objective of this study was to investigate the effect of incorporation of polyethylenimine (PEI) on rAAV-mediated insulin gene therapy in vitro and in vivo.

Methods: Recombinant AAV vector, harboring the furin-mutated human insulin and enhanced green fluorescent protein (EGFP) genes, was constructed.

The immunogenicity-enhancing effect of emulsion vaccine adjuvants is independent of the dispersion type and antigen release rate--a revisit of the role of the hydrophile-lipophile balance (HLB) value.

Effective antigen delivery is one of the most important issues in vaccine development. It has been suggested that adjuvant action results from a depot effect by prolonging the duration of the interaction between antigen and cells, and thus is related to the antigen-releasing properties of emulsion adjuvants. The objective of this study was to investigate the effect of the dispersion properties of emulsion-type vaccine adjuvants on the immune response with the aim of optimizing vaccine adjuvant formulation.

The apoptotic and necrotic effects of tomatine adjuvant.

Tomatine adjuvant, consisting of tomatine, n-octyl-beta-d-glucopyranoside (OGP), phosphatidylethanolamine and cholesterol is unique in that when combined with soluble protein antigen it elicits a cytotoxic T lymphocyte (CTL) response in immunized animals. The mechanisms underlying this property are unknown. In an attempt to understand how tomatine activates cellular immunity, we examined its potential to induce apoptosis.

Cell death induced by vaccine adjuvants containing surfactants.

Many vaccine adjuvants contain surface-active agents, but the immunological roles played by these components have been essentially ignored. The objective of this study was to examine possible apoptotic and necrotic effects of the surface-active agents, Pluronic L121 and Tween 80, which are components of L121-adjuvant (a formulation we synthesized with the aim of representing several commercially produced adjuvants), on EL4 lymphoma cells. Cell viability and cytolytic effects were analyzed using the MTT and LDH release assays, and the distribution of cells in different stages of the cell cycle after treatment with these agents was analyzed by propidium iodide (PI) staining and flow cytometry.

Induction of cell death by saponin and antigen delivery.

Purpose: Saponin is the major component in the formation of immune stimulating complex (ISCOM), a potent adjuvant able to induce both humoral and cellular immune reactions. The immunogenicity induced by saponin, however, has been unclear. The objective of this study was to investigate the apoptotic and necrotic effects induced by saponin in ELA mouse lymphoma cells, expected to be a possible mechanism of the cytotoxic T-lymphocyte (CTL) effect elicited by the ISCOM.

Differential requirements for CTL generation by novel immunostimulants: APC tropism, use of the TAP-independent processing pathway, and dependency on CD80/CD86 costimulation.

A major drawback of subunit vaccines is their inability to generate cytolytic T lymphocytes (CTL), a deficit attributed to segregation of the class I and class II antigen-processing pathways. We sought to understand processes involved in CTL induction by three proprietary adjuvants: Tomatine, PROVAX, and a synthesized glycolipid (Glc-N-(8/16), Glycolipid). We used in vivo models to investigate antigen uptake, macrophage involvement, TAP-independent processing, and costimulatory molecule dependencies.

Incorporation of calcium phosphate enhances recombinant adeno-associated virus-mediated gene therapy in diabetic mice.

Background: Increased efficiency of transgene expression is desired for virus-mediated gene delivery. In the present study, we examined the effect of calcium phosphate (CaPi) on recombinant adeno-associated virus (rAAV)-mediated insulin therapy in diabetic animals.

Methods: The rAAV vector, rAAV.

The ultrastructure of tomatine adjuvant.

The tomatine adjuvant, consisting of tomatine, n-octyl-beta-D-glucopyranoside, phosphatidylethanolamine, cholesterol, and ovalbumin, has recently been shown to potentiate the immunogenicity of protein antigen and elicit cytotoxic T-lymphocyte responses in immunized animals. The physicochemical properties of tomatine adjuvant have not been characterized. The aim of this study was to examine the microstructure of this complex formulation, as directly related to its physicochemical properties.

Glucose-modulated transgene expression via recombinant adeno-associated virus.

Purpose: The objective of this study was to examine glucose modulated reporter gene expression via recombinant adeno associated viral vectors both in vitro and in vivo.

Methods: Huh7 human hepatoma cells were transduced by recombi nant adeno-associated virus (rAAV) vectors containing the luciferase gene under control of the rat insulin I gene promoter and a cytomegalovirus immediate-early promoter driving-enhanced green fluores cence protein gene. The reporter gene expression was evaluated by glucose stimulation either in the absence or presence of insulin se cretagogues, including phorbol-12-myristate-13-acetate, dibutyryl cy clic AMP, and forskolin.