Protective effect of an egg yolk-derived immunoglobulin (IgY) against Prevotella intermedia-mediated gingivitis.

AIMS: To investigate the effects of an egg yolk-derived immunoglobulin (IgY) specific to Prevotella intermedia in vitro and in vivo. METHODS AND RESULTS: An IgY specific to P. intermedia was produced by immunizing hens with formaldehyde-inactivated P. intermedia and showed high titres when subjected to an ELISA. The obtained IgY inhibited the growth of P. intermedia in a dose-dependent manner at concentrations from 1 to 20 mg ml(-1) in Center for Disease Control and Prevention liquid medium. Forty rats were challenged with P. intermedia on gingivae and then randomly divided into four groups, which were syringed respectively with phosphate-buffered saline, 1 mg ml(-1) of tinidazole, 20 mg ml(-1) of nonspecific IgY and 20 mg ml(-1) of the IgY specific to P. intermedia at a dosage of 300 µl per day. Gingival index (GI), plaque index (PI), bleeding on probing (BOP), counts of white blood cell (WBC) and histopathological slide of the gums were measured after treatment for 15 days. The gingivitis rats treated with the IgY specific to P. intermedia showed significantly decreased GI, PI, BOP and WBC (P < 0·05). Gum histopathology of the treated rats demonstrated a superior protective effect of the specific IgY on P. intermedia-mediated gingivitis. CONCLUSIONS: A new immunoglobulin specific to P. intermedia was developed from egg yolk. This specific IgY can dose-dependently inhibit the growth of P. intermedia and protect rats from gingivitis induced by P. intermedia. SIGNIFICANCE AND IMPACT OF THE STUDY: The new IgY has potential for the treatment of P. intermedia-mediated gingivitis.

Isolation and identification of metabolites of osthole in rats.

Osthole (Ost), one of the major components of Cnidium monnieri (L.) Cusson, is had the structure of an isopentenoxy-coumarin with a range of pharmacological activities. In the present study, the metabolism of Ost in male Sprague-Dawley rats was investigated by identifying Ost metabolites excreted in rat urine. Following an oral dose of 40 mg/kg Ost, 10 phase I and 3 phase II metabolites were isolated from the urine of rats, and their structures identified on the basis of a range of spectroscopic data, including 2D-NMR techniques. These metabolites were fully characterized as 5'-hydroxyl-osthole (M-1), osthenol (M-2), 4'-hydroxyl-osthole (M-3), 3, 5'-dihydroxyl-osthole (M-4), 5'-hydroxyl-osthenol (M-5), 4'-hydroxyl-2', 3'-dihydro-osthenol (M-6), 4'-hydroxyl-osthenol (M-7), 3, 4'-dihydroxyl-osthole (M-8), 2', 3'-dihydroxyl-osthole (M-9), 5'-hydroxyl-2', 3'-dihydroosthole (M-10), osthenol-7-O-ß-D-glucuronide (M-11), osthole-4'-O-ß-D-glucuronide (M-12) and osthole-5'-O-ß-D-glycuronate (M-13). This is the first identification of M-1, M-3 to M-13 in vivo. On the basis of the metabolites profile, a possible metabolic pathway for Ost metabolism in rats has been proposed. This is the first systematic study on the phases I and II metabolites of 8-isopentenoxy-coumarin derivative.

Three-dimensional analysis of autologous costal cartilage in Asian secondary unilateral cleft rhinoplasty.

This study describes a surgical technique for secondary unilateral cleft rhinoplasty using autologous costal cartilage grafts. The grafts were designed preoperatively and analysed three-dimensionally in 15 Asian patients using a photogrammetric camera. Detailed measurements of the nasal anatomy were taken both preoperatively and postoperatively; the same measurements were also taken from the pre-planned images of the anticipated result. When compared to the preoperative measurements, the postoperative three-dimensional outcome analysis revealed several statistically significant improvements in the nasal appearance: nasal dorsal length (P < 0.001), nasal column height (P = 0.001), nasal column width (P = 0.002), nasal lobule height (P = 0.008), cleft side nostril height (P < 0.001) and width (P < 0.001), columella-labial angle (P = 0.001), and nasal tip projection to nasal dorsum length ratio (NTP/NDL) (P = 0.001). Conversely, the comparison of the postoperative and preoperative design measurements showed mostly no statistically significant differences. Thus, utilizing autologous costal cartilage is a reliable approach with predictable and consistent results in secondary cleft rhinoplasty.

Efficacy of specific IgY for treatment of lipopolysaccharide-induced endotoxemia using a mouse model.

AIMS: To estimate the efficacy of specific egg yolk immunoglobulin (IgY) for the treatment of lipopolysaccharide (LPS)-induced endotoxemia using a mouse model. METHODS AND RESULTS: Specific IgY was obtained from the yolk of hens immunized with formaldehyde-killed Escherichia coli O111 and showed a high binding activity to LPS when subjected to an ELISA. Endotoxemia was induced in mice by intraperitoneal injection of LPS at a dose of 20 mg kg(-1) for measuring survival rate and 10 mg kg(-1) for cytokine measurement. The survival rate of mice treated with 200 mg kg(-1) specific IgY or 5 mg kg(-1) dexamethasone was 70% while none of the mice in the normal saline-treated group survived more than 7 days. Specific IgY significantly (P < 0.05) decreased tumour necrosis factor-α (TNF-α) level and increased interleukin-10 (IL-10) level in the serum of endotoxemia mice. Specific IgY had less of an effect on TNF-α than dexamethasone, while its effect on increasing IL-10 was stronger than dexamethasone. Haematoxylin and eosin-stained sections indicated that IgY attenuated the damage to the lung and liver observed in mice with endotoxemia. CONCLUSIONS: The specific IgY increased the survival rate of mice with endotoxemia induced by LPS, down-regulated TNF-α and up-regulated IL-10 in serum and attenuated the extent of damage to the lung and liver. SIGNIFICANCE AND IMPACT OF THE STUDY: The specific IgY has potential for the treatment of LPS-induced endotoxemia.

Anti-breast cancer activity of resveratrol encapsulated in liposomes.

Resveratrol (RES) is a naturally occurring and effective drug for tumor prevention and treatment. However, its low levels of aqueous solubility, stability, and poor bioavailability limit its application, especially when used as a free drug. In this study, RES was loaded into peptide and sucrose liposomes (PSL) to enhance the physico-chemical properties of RES and exploit RES delivery mediated by liposomes to effectively treat breast cancer. RES loaded PSL (the complex: PSL@RES) were stable, had a good RES encapsulation efficiency, and prolonged RES-release in vitro. PSL@RES was exceptionally efficient for inhibiting the growth of cancer cells, as the IC50 of PSL@RES in MCF-7 cells was found to be only 20.89 µmol L-1. The therapeutic efficacy of PSL@RES was evaluated in mice bearing breast cancer. The results showed that PSL@RES at a dosage of 5 mg kg-1 was more effective than 10 mg kg-1 free RES, and PSL@RES inhibited tumor growth completely at a dosage of 10 mg kg-1. PSL@RES induced apoptosis in breast tumor by upregulation of p53 expression. This then downregulated Bcl-2 and upregulated Bax, thereby inducing Caspase-3 activation. More importantly, encapsulation of RES within peptide liposomes greatly reduced the toxicity of free RES to mice. Overall, the simple formulation of liposomal nanocarriers of RES developed in this study produces satisfactory outcomes to encourage further applications of liposomal carriers for the treatment of breast cancer.

Characterization of specific egg yolk immunoglobulin (IgY) against mastitis-causing Staphylococcus aureus.

AIMS: To evaluate the in vitro activity of egg yolk immunoglobulin (IgY) against mastitis-causing Staphylococcus aureus. METHODS AND RESULTS: Specific IgY was produced by immunizing hens with formaldehyde-killed Staph. aureus, using a bacterial strain known to cause mastitis. The IgY, of 94% purity, was obtained from yolks by water dilution, salt precipitations, ultrafiltration and gel filtration. ELISA indicated that the IgY produced was specific to the antigen and five Staph. aureus isolates obtained from mastitic cows. The growth of Staph. aureus was inhibited by specific IgY at concentrations from 1 to 10 mg ml(-1) in a dose-dependent manner. The phagocytosis of Staph. aureus by milk macrophages was enhanced in the presence of specific IgY with the highest phagocytic percentage being 30% higher than that without IgY (P < 0.05). CONCLUSIONS: The specific IgY against mastitis-causing Staph. aureus inhibited the growth of Staph. aureus and enhanced the phagocytosis of Staph. aureus by milk macrophages. SIGNIFICANCE AND IMPACT OF THE STUDY: Specific IgY would be a potential treatment for bovine mastitis.

Replacement of quaternary ammonium headgroups by tri-ornithine in cationic lipids for the improvement of gene delivery in vitro and in vivo.

Headgroups in cationic lipids play very important roles in determining transfection efficiency and toxicity in gene delivery. To better understand the influence of headgroups on gene delivery, a tri-peptide-based lipid was synthesized, wherein the usual quaternary ammonium was replaced by a tri-peptide. Though both the tri-peptide-based lipid (DAO3) and the quaternary ammonium-based lipid (DDCTMA) successfully mediated gene transfection, DAO3 was superior to DDCTMA in both in vitro and in vivo studies. Following their preparation into liposomes, the particle size, zeta potential, and DNA-binding capacity of the liposomes and lipoplexes were characterized to evaluate the efficiency of DAO3 compared to DDCTMA with regard to gene interactions. The expression of luciferase from pDNA mediated by DAO3 was 2-fold greater than than that with DDCTMA in Hep-2 cells, and DAO3/siRNA lipoplexes could silence about 60% luciferase in A549 cancer cells expressing firefly luciferase. DAO3/Luc-siRNA treatment exhibited 3-fold the efficiency of DDCTMA/Luc-siRNA in terms of in vivo luciferase RNAi with the bare density ratio of 0.54 at 48 h. Furthermore, DAO3 could mediate IGF-1R siRNA to inhibit tumor growth through silencing the expression of the IGF-1R protein, whereas DDCTMA showed nearly no effects. Most importantly, DAO3 had no obvious toxicity in vitro and in vivo, due to the biocompatibility of the peptide headgroups. In conclusion, these results demonstrated that the replacement of the quaternary ammonium headgroup by tri-ornithine may increase transfection efficiency and decrease toxicity.