Enhanced Efficacy of Immunization with a Foot-and-Mouth Disease Multi-Epitope Subunit Vaccine Using Mannan-Decorated Inulin Microparticles

BACKGROUND@#Despite the many advantages of recombinant subunit vaccines, they have critical weaknesses that include a low efficacy for promoting cellular and humoral immune responses against antigens because of their poor immunogenicity, and a rapidly cleared properties as a result of proteolytic enzymes in the body. To circumvent these problems, we developed mannan-decorated inulin acetate microparticles (M-IA MPs) that functioned as carriers and adjuvants for immunization with the recombinant foot-and-mouth disease multi-epitope subunit vaccine (M5BT). @*METHODS@#The M5BT-loaded M-IA MPs were obtained by a double-emulsion solvent-evaporation method. Their properties including morphology, size and release ability were determined by field emission scanning electron microscope, dynamic light-scattering spectrophotometer and spectrophotometer. To assess the immunization efficacy of the MPs, mice were immunized with MPs and their sera were analyzed by ELISA. @*RESULTS@#The M-IA MPs obtained by a double-emulsion solvent-evaporation method were spherical and approximately 2–3 µm, and M5BT was encapsulated in the M-IA MPs. The M5BT-loaded M-IA MPs showed higher antigen-specific IgG, IgG1, IgG2a and anti-FMDV antibodies than the M5BT-loaded IA MPs and the Freund’s adjuvant as a control. @*CONCLUSION@#The M-IA MPs showed a powerful and multifunctional polymeric system that combined two toll-like receptor agonists compared to the conventional adjuvant.

Enhanced Efficacy of Immunization with a Foot-and-Mouth Disease Multi-Epitope Subunit Vaccine Using Mannan-Decorated Inulin Microparticles

BACKGROUND@#Despite the many advantages of recombinant subunit vaccines, they have critical weaknesses that include a low efficacy for promoting cellular and humoral immune responses against antigens because of their poor immunogenicity, and a rapidly cleared properties as a result of proteolytic enzymes in the body. To circumvent these problems, we developed mannan-decorated inulin acetate microparticles (M-IA MPs) that functioned as carriers and adjuvants for immunization with the recombinant foot-and-mouth disease multi-epitope subunit vaccine (M5BT). @*METHODS@#The M5BT-loaded M-IA MPs were obtained by a double-emulsion solvent-evaporation method. Their properties including morphology, size and release ability were determined by field emission scanning electron microscope, dynamic light-scattering spectrophotometer and spectrophotometer. To assess the immunization efficacy of the MPs, mice were immunized with MPs and their sera were analyzed by ELISA. @*RESULTS@#The M-IA MPs obtained by a double-emulsion solvent-evaporation method were spherical and approximately 2–3 µm, and M5BT was encapsulated in the M-IA MPs. The M5BT-loaded M-IA MPs showed higher antigen-specific IgG, IgG1, IgG2a and anti-FMDV antibodies than the M5BT-loaded IA MPs and the Freund’s adjuvant as a control. @*CONCLUSION@#The M-IA MPs showed a powerful and multifunctional polymeric system that combined two toll-like receptor agonists compared to the conventional adjuvant.

Enhancement of antigen-specific humoral immune responses and protein solubility through conjugation of bacterial flagellin, Vibrio vulnificus FlaB, to the N-terminus of porcine epidemic diarrhea virus surface protein antigen S0

Porcine epidemic diarrhea (PED) is a highly contagious enteric swine disease. The large economic impact of PED on the swine industry worldwide has made the development of an effective PED vaccine a necessity. S0, a truncated region of the porcine epidemic diarrhea virus (PEDV) spike protein, has been suggested as a candidate antigen for PED subunit vaccines; however, poor solubility problems when the protein is expressed in Escherichia coli, and the inherent problems of subunit vaccines, such as low immunogenicity, remain. Flagellin has been widely used as a fusion partner to enhance the immunogenicity and solubility of many difficult-to-express proteins; however, the conjugation effect of flagellin varies depending on the target antigen or the position of the fusion placement. Here, we conjugated flagellin, Vibrio vulnificus FlaB, to the N- and C-termini of S0 and evaluated the ability of the fusion to enhance the solubility and immunogenicity of S0. Flagellin conjugation in the presence of the trigger factor chaperone tig greatly improved the solubility of the fusion protein (up to 99%) regardless of its conjugation position. Of importance, flagellin conjugated to the N-terminus of S0 significantly enhanced S0-specific humoral immune responses compared to other recombinant antigens in Balb/c mice. The mechanism of this phenomenon was investigated through in vitro and in vivo studies. These findings provide important information for the development of a novel PED vaccine and flagellin-based immunotherapeutics.

Oral Immunization of FMDV Vaccine Using pH-Sensitive and Mucoadhesive Thiolated Cellulose Acetate Phthalate Microparticles

Several barriers such as gastric pH, enzymatic degradation and rapid transit should be overcome to orally deliver antigens for taking up by epithelial microfold cells in Peyer's patches of small intestine. To solve the above mentioned problems, we designed pH-sensitive and mucoadhesive polymeric microparticles (MPs) prepared by double emulsion technique using cellulose acetate phthalate (CAP) to enhance immune response of foot-and-mouth disease (FMD) virus (FMDV) subunit vaccine. Thiolation of CAP improved mucoadhesive property of CAP to prolong the MPs transit time through the gastrointestinal tract. Thiolated CAP (T-CAP) also slowed down antigen release in acidic pH of stomach but released more antigens in neutral pH of small intestine due to the pH-sensitivity of the T-CAP. Oral immunization of a chimerical multi-epitope recombinant protein as the FMD subunit vaccine via T-CAP MPs effectively delivered the vaccine to Peyer's patches eliciting mucosal IgA response. It will make a step forward into a promising oral subunit vaccine development in livestock industry.

Evaluation of Influence of Individual Facial Aesthetic Subunits on the Congnition of Facial Attractiveness in Public

PURPOSE: Authors tried to analyze the influence of individual facial aesthetic subunits on the cognition of facial attractiveness in public and suggest a mathematical model which explain the facial attractiveness. METHODS: Independent facial aesthetic subunits are extracted from facial photographs from three women(11 frontal and 7 lateral aesthetic subunits). Each facial subunits of three women are rated in terms of relative rank by 164 peoples(68 man and 96 woman, average age was 32.4, and ranged +/- 9.8 years). chi-square-test and categorical regression analysis were performed. RESULTS: There was no difference in the aesthetic preference in terms of ages or sexes in large. Beautification of individual aesthetic subunits can predict the overall facial attractiveness up to 42.1% in frontal face(Adjusted R2=0.421, F=6.39, p=0.000<0.05) and 22.7% in lateral face (Adjusted R2=0.227, F=4.42, p=0.000<0.05). Aesthetic appearance of eyes(p=0.001), upper face(p=0.034) in frontal face and midface(p=0.000) in lateral face are statistically important factors in the cognition of facial attractiveness. CONCLUSION: Authors experimently proved that harmony and balance among facial aesthetic subunits are the most important factors, in embarking on facial aesthetic plastic surgery, for better enhancement of facial attractiveness.

MACS Lift on Asian: Consideration on Third Purse String Suture and Malar Region

The lifting of gravitationally relaxed cheek could make Asian patients to be dissatisfied with the surgical results due to the protrusion of zygoma. When the MACS-lift was applied as the facial lifting technique to counter the effects of aging in the mid-facial area of Asian patients, the third purse-string suture that lifts the malar fat pad was applied on most of the patients with normally-shaped zygoma. The simple MACS lift and extensive MACS lift were conducted for 3 patients and 18 patients each, respectively. The survey after 6 months from the surgery showed "satisfied" in 15 patients, "very satisfied" in 3 patients and "no difference" in 3 patients. The other two plastic surgeons evaluated the results after 6 months from the surgery. In the experience of the authors, the third purse-string suture of the MACS lift was effective in the elevation of the malar fat pad. Considering the Asian facial characteristics of wide and protruded zygoma, the fullness of the zygomatic area has been an issue for concern, however, patients who wanted to have a younger-looking face are now satisfied with this appearance due to changes in aesthetic standards in recent years.

Various Use of Perforator Pedicled Flap: First Choice instead of Free Flap and Pedicled Flap

PURPOSE: Because of good blood supply of the pedicled perforator flap and its advantage of not requiring vessel anastomosis compared to pedicled flap, it is widely used recently. The authors intended to report the results of various pedicled perforator flaps which have been performed to reconstruct the soft tissue defects and the utility of the flap. METHODS: The study was conducted for 12 cases of pedicled perforator flap which were performed at the plastic surgery department of the current hospital from the period of June, 2005 to August, 2008. Four patients were male and eight patients were female and their age was ranged from 22 to 74 years old with mean age of 42.6 years old. The sites were 1 case on face, 3 cases on chest, 3 cases on back, 4 cases on coccyx, and 1 case on foot. RESULTS: The defect sites of all patients were successfully reconstructed by using the pedicled perforator flap. Although most of the flaps revealed congestion at the early stage after the surgery, they were recuperated within few days. One case of skin flap was reported to be partially necrotized in old age woman who has no reliable perforator. Other than that, all defects were covered successfully and acceptable aethetically. CONCLUSION: As stated in above, the pedicled perforator flap has many useful advantages than the conventional pedicled flap and various free flap. Unless free flap must be required, the use of pedicled perforator flap is recommended by first choice for soft tissue coverage.