Genome-wide identification of potential odontogenic genes involved in the dental epithelium-mesenchymal interaction during early odontogenesis.

BACKGROUND: Epithelium-mesenchymal interactions are involved in odontogenic processes. Previous studies have focused on the intracellular signalling regulatory network in tooth development, but the functions of extracellular regulatory molecules have remained unclear. This study aims to explore the gene profile of extracellular proteoglycans and their glycosaminoglycan chains potentially involved in dental epithelium-mesenchymal interactions using high-throughput sequencing to provide new understanding of early odontogenesis. RESULTS: Whole transcriptome profiles of the mouse dental epithelium and mesenchyme were investigated by RNA sequencing (RNA-seq). A total of 1,281 and 1,582 differentially expressed genes were identified between the dental epithelium and mesenchyme at E11.5 and E13.5, respectively. Enrichment analysis showed that extracellular regions and ECM-receptor interactions were significantly enriched at both E11.5 and E13.5. Polymerase chain reaction analysis confirmed that the extracellular proteoglycan family exhibited distinct changes during epithelium-mesenchymal interactions. Most proteoglycans showed higher transcript levels in the dental mesenchyme, whereas only a few were upregulated in the epithelium at both stages. In addition, 9 proteoglycans showed dynamic expression changes between these two tissue compartments. Gpc4, Sdc2, Spock2, Dcn and Lum were expressed at higher levels in the dental epithelium at E11.5, whereas their expression was significantly higher in the dental mesenchyme at E13.5, which coincides with the odontogenic potential shift. Moreover, the glycosaminoglycan biosynthetic enzymes Ext1, Hs3st1/5, Hs6st2/3, Ndst3 and Sulf1 also exhibited early upregulation in the epithelium but showed markedly higher expression in the mesenchyme after the odontogenic potential shift. CONCLUSION: This study reveals the dynamic expression profile of extracellular proteoglycans and their biosynthetic enzymes during the dental epithelium-mesenchymal interaction. This study offers new insight into the roles of extracellular proteoglycans and their distinct sulfation underlying early odontogenesis.

Marfan Syndrome With Recurrent Lower Left Posterior Toothache as the First Symptom: A Report on a Rare Case.

Marfan syndrome (MFS) is a life-threatening connective tissue disorder that affects multiple organs and systems. We report a case of MFS with recurrent lower left posterior toothache as the first symptom. A 23-year-old Asian man walked into the dental emergency room with a chief complaint of recurrent spontaneous and intermittent toothache in his lower left posterior tooth region, mimicking acute symptomatic pulpitis. He self-reported a relatively healthy medical status without any hereditary disease. However, his disproportionately elongated body structure, high myopia, and positive wrist sign were immediately recognizable. Although there were no remarkable findings on the dental examination, pectus carinatum deformity and abnormal blood pressure were later detected. He was immediately referred to a cardiologist in a medical hospital. The timely diagnosis of MFS and early surgical intervention helped to avoid severe lethal consequences. The symptoms of toothache completely resolved after surgery.

Coating function and stabilizing effects of surface layer protein from Lactobacillus acidophilus ATCC 4356 on liposomes.

Surface layer proteins (SLPs) are crystalline arrays in the outermost layer of cell envelope in many archaea and bacteria. SLPs subunits have the ability to reassemble on the surface of lipid layers. In this work, the SLP from Lactobacillus acidophilus ATCC 4356 was extracted and reassembled on the surface of positively charged liposomes composed of dipalmitoyl phosphatidylcholine, cholesterol and octadecylamine. Zeta potentials and particle size were determined to describe the adsorption process of SLP on liposomes. The liposomes completely coated with SLP were observed by transmission electron microscope. To investigate the stabilizing effects of SLP on liposomes, carboxyfluorescein (CF) was encapsulated and its leakage was determined as an evaluation index. The results showed that the L. acidophilus ATCC 4356 SLP significantly (P < 0.05) increased the stability of the liposomes in the course of thermal challenge. Furthermore, SLP was able to reduce the aggregation of liposomes in serum. Storage stability of liposomes was performed at 25 °C, 4 °C and -20 °C for 90 days. And the SLP-coated liposomes released less CF than the control liposomes during storage at the three evaluated temperatures. Our findings extended the application field of Lactobacillus SLPs and introduced a novel nanocarrier system with good chemical stability.

Integration of purification with immobilization of Candida rugosa lipase for kinetic resolution of racemic ketoprofen.

The two processes for the partial purification and for the immobilization of a crude lipase preparation (Candida rugosa Lipase OF) have been successfully integrated into one by simple adsorption of the enzyme onto a cation ion exchanger resin (SP-Sephadex C-50) at pH 3.5. Due to selective removal of the unfavorable lipase isoenzyme (L1), the enzyme components (mainly L2 and L3) that are tightly fixed on the resin displayed a significantly improved enantioselectivity (E value: 50 versus 13 with addition of Tween-80) in the biocatalytic hydrolysis of 2-chloroethyl ester of rac-ketoprofen. The activity yields of the immobilized lipase were 48 and 70%, respectively when emulsified and non-emulsified substrates were employed for enzyme assay. Moreover, the concentration of Tween-80 was found to be a factor affecting the lipase enantioselectivity. By using such an immobilized enzyme as biocatalyst, the process for preparing enantiopure (S)-ketoprofen becomes simpler and more practical as compared with the previously reported procedures and the product was obtained with >94% ee at 22.3% conversion in the presence of an optimal concentration (0.5 mg/ml) of Tween-80 at pH 3.5. Furthermore, the operational stability of the immobilized biocatalyst was examined in different types of reactors. In an air-bubbled column reactor, the productivity was much higher than that in a packed-bed column reactor, in spite of a slightly lower stability. Under optimal conditions, the air-bubbled column reactor could be operated smoothly for at least 350 h, remaining nearly 50% activity.

A novel non-hydrolytic protein from Pseudomonas oryzihabitans enhances the enzymatic hydrolysis of cellulose.

Several kinds of protein such as the expansin, expansin-like proteins and LPMOs (lytic polysaccharide monooxygenases) are known to exert enhancement effects on cellulase activity. In this study, a novel cellulase synergistic protein named POEP1 was purified from the culture filtrate of Pseudomonas oryzihabitans CGMCC 6169, and was homogeneous on SDS-PAGE with a molecular weight of 60kDa. Mass spectrometry analysis indicated that it was an unknown protein without sequence similarity to the expansin and expansin-like proteins. Evaluation of the enzymatic hydrolysis of filter paper revealed that POEP1 had no cellulase activity but displayed high synergistic activity of 364% at a cellulase concentration of 0.1FPU/g of filter paper. When a mixture containing 0.6FPU cellulase and 700µg POEP1 per g of cellulose was evaluated, the maximal sugar yield was achieved, which was 2.2-fold greater than that with the cellulase alone. POEP1 was found to have functional similarity to the expansin and expansin-like proteins, which could decrease both the hydrogen-bond intensity and crystallinity, and cause the filter paper disruption. This study provided evidence for the existence of novel bacterial proteins in nature serving the same function as expansin and expansin-like proteins.

Enzymatic saccharification of sugarcane bagasse by N-methylmorpholine-N-oxide-tolerant cellulase from a newly isolated Galactomyces sp. CCZU11-1.

Based on the enrichment culture strategy, a novel N-methylmorpholine-N-oxide (NMMO)-tolerant cellulase-producing strain Galactomyces sp. CCZU11-1 was isolated from soil samples. After the optimization of culture condition, the highest FPA (13.4 U/mL) and CMCase (24.5 U/mL) were obtained. In both culture and reaction media containing NMMO 25% (w/v), the cellulase from Galactomyces sp. CCZU11-1 still had good activity. Furthermore, high saccharification rate was obtained in aqueous-NMMO media. Moreover, the fermentability of the hydrolyzates, obtained after enzymatic in situ saccharification of the NMMO-pretreated sugarcane bagasse, was evaluated using Saccharomyce scerevisiae. In conclusion, Galactomyces sp. CCZU11-1 is a promising candidate as high NMMO-tolerant cellulase producer and has potential application in future.

[Screening and identification of microorganisms for decolorization of molasses spent wash].

Microorganisms were screened from the natural environment for decolorization of molasses spent wash, and the isolated strains were then employed in the treatment of actual wastewater. The primary screening was carried out on agar plates supplemented with synthesized melanoidin as the target substrate, since melanoidin is one of the most refractory pigments in wastewater. Promising microorganisms were further selected through secondary screening by decolorization of untreated actual wastewater in shaking flask cultures. Gel filtration chromatography was used to determine the molecular weight distribution of pigments in molasses spent wash before and after decolorization. A strain named A5P1 was isolated from the soil samples collected, showing a good ability of decolorizing molasses spent wash, and was later identified as Aspergillus flavus by morphology and ITS sequence analysis. Experimental study of factors affecting the decolorization performance of strain A5P1 gave the optimal conditions as follows: 4.3 x 10(4) mL(-1) of inoculum size, medium with initial pH of 4.5 and cultivation at 39 degrees C. It could decolorize 53.0% of the pigments in the untreated molasses spent wash and decreased 80% of chemical oxygen demand after four-day incubation. The result of gel filtration chromatography demonstrated that both the large and small molecular weight fractions of pigments in the molasses spent wash could be removed by strain A5P1. Based on the measurement of enzyme activities, at least three different kinds of enzymes, i. e. the enzyme with H2O2-producing activity, laccase and manganese peroxidase were involved in the decolorization process. Therefore, the decolorization mechanism of strain A5P1 was preliminarily considered to be mainly biodegradation, with bioadsorption as a minor reaction.

[Effect of periodontitis on circulating C-reactive protein in type 2 diabetes patients].

OBJECTIVE: To investigate the effect of periodontal infection on circulating C-reactive protein (CRP) in type 2 diabetes patients. METHODS: 32 diabetes patients with advanced periodontitis participated in this study. They were compared to a group of 32 diabetes patients without periodontal disease, who were mathed with regard to age (+/- 3 years), gender and body mass index (+/- 1 kg/m2). The concentration of CRP on circulation was measured by ELISA. RESULTS: Significant difference was found in the level of CRP and the percentage of subjects with elevated CRP levels > or = 3 mg/L on circulation between the two groups(P < 0.05). CONCLUSION: Periodontal infection results in higher circulating CRP in type 2 diabetes patients. This elevated inflammatory factor may exacerbate insulin resistance and increase the risk for great vessels complications of diabetes mellitus.