Lactobacillus crispatus (L. crispatus) ZJ001 is highly adhesive to epithelial cells and expresses S-layer proteins. In this study, S-S-layer layer genes were sequenced and expressed in E. coli to characterize the function of proteins with this particular strain. L. crispatus ZJ001 harbored two S-layer genes slpA and slpB, and only slpA gene was expressed in the bacterium, as revealed by RT-PCR and immunoassays. The mature SlpA showed 47% amino acid sequence identity to SlpB. The SlpA and SlpB of L. crispatus ZJ001 were highly homologous at the C-terminal region to other Lactobacillus S-layer proteins, but were substantially variable at N-terminal and middle regions. Electron microscopic analysis indicated that His-slpA expressed in E. coli was able to form a sheet-like structure similar to the natural S-layer, but His-slpB formed as disc-like structures. In the cell binding experiments, HeLa cells were able to bind to both recombinant His-slpA and His-slpB proteins to the extent similar to the natural S-layer. The cell binding domains remain mostly in the N-terminal regions in SlpA and SlpB, as shown by high binding of truncated peptides SlpA2-228 and SlpB2-249. Our results indicated that SlpA was active and high binding to HeLa cells, and that the slpA gene could be targeted to display foreign proteins on the bacterial surface of ZJ001 as a potential mucosal vaccine vector.
Lee, Eun-Young;Jun, Sul-Gi;Wright, Robert F.;Park, Eun-Jin
The Journal of Advanced Prosthodontics
/
v.7
no.1
/
pp.69-75
/
2015
PURPOSE. To compare the shear bond strength of various veneering materials to grade II commercially pure titanium (CP-Ti). MATERIALS AND METHODS. Thirty specimens of CP-Ti disc with 9 mm diameter and 10 mm height were divided into three experimental groups. Each group was bonded to heat-polymerized acrylic resin (Lucitone 199), porcelain (Triceram), and indirect composite (Sinfony) with 7 mm diameter and 2 mm height. For the control group (n=10), Lucitone 199 were applied on type IV gold alloy castings. All samples were thermocycled for 5000 cycles in $5-55^{\circ}C$ water. The maximum shear bond strength (MPa) was measured with a Universal Testing Machine. After the shear bond strength test, the failure mode was assessed with an optic microscope and a scanning electron microscope. Statistical analysis was carried out with a Kruskal-Wallis Test and Mann-Whitney Test. RESULTS. The mean shear bond strength and standard deviations for experimental groups were as follows: Ti-Lucitone 199 ($12.11{\pm}4.44$ MPa); Ti-Triceram ($11.09{\pm}1.66$ MPa); Ti-Sinfony ($4.32{\pm}0.64$ MPa). All of these experimental groups showed lower shear bond strength than the control group ($16.14{\pm}1.89$ MPa). However, there was no statistically significant difference between the Ti-Lucitone 199 group and the control group, and the Ti-Lucitone 199 group and the Ti-Triceram group. Most of the failure patterns in all experimental groups were adhesive failures. CONCLUSION. The shear bond strength of veneering materials such as heat-polymerized acrylic resin, porcelain, and indirect composite to CP-Ti was compatible to that of heatpolymerized acrylic resin to cast gold alloy.
Background: Several biochemical mediators, such as substance P, calcitonin gene-related peptide (CGRP) and prostaglandin $E_2$, have been demonstrated to be involved in herniated or degenerated disc-induced radiculopathy. The authors tested the hypothesis that these mediators would existed in the epidural space of humans. Methods: Thirty nine patients were divided into two groups; 27 patients, who were diagnosed with spinal stenosis (stenosis group), and 12 scheduled for epidural anesthesia, without a history of back pain (control group). Under fluoroscopic guidance, an epidural catheter was introduced through the caudal space and placed into the anterior and posterior spaces, up to and around the epidural adhesive area, in the stenosis group. In the control group, the catheter was placed into the posterior epidural space through the L3-4 or L4-5 intervertebral space. Epidural irrigation was performed with 10 ml of saline, via an epidural catheter. Aspirated lavage fluid was collected, and the concentrations of biochemical mediators (substance P, CGRP and prostaglandin $E_2$) measured using an enzyme immunoassay kit. Results: Substance P, CGRP and prostaglandin $E_2$ were detected in all the epidural lavage fluids from both groups. The concentrations of substance P and prostaglandin $E_2$ in the stenosis group were higher than those of the control (P < 0.05). However, there was no difference in the CGRP levels between the two groups. In the stenosis group, the concentrations of these three mediators in the anterior epidural space were no different to those in the posterior space. Conclusions: These results suggest that biochemical mediators, such as substance P and prostaglandin $E_2$, in the epidural space might be partly involved in pain mechanism associated with spinal stenosis.
The process of embryogenesis and larval development of the asteroid sea star Asterias amurensis $(U{\ddot{u}}tken)$ was observed, with special attention paid to morphological change and larval duration. In reproductive season, mature sea stars were collected under floating net cages, located in Tongyeong, southern Korea. The mature eggs are $138\;{\mu}m$ in average diameter, semi-translucent and orange in color, sperms in good condition appear light cream to white-gray in color. Embryos develop through the holoblastic equal cleavage stage and a wrinkled blastula stage that lasts about 9 hours after fertilization. Gastrulae bearing an expanded archenteron hatch from the fertilization envelope 22 hours after fertilization. At the end of gastrulation, rudiments of the left and right coelom are formed. By day 2, larvae possess complete alimentary canal and begin to feed. At this stage, the larva is called early bipinnaria. In 6-day-old larvae, the pre- and post- oral ciliated bands form complete circuits and the bipinnarial processes start to develop. By day 12, the lateral and anterior projection of the larval wall processes along the ciliated bands begins to thicken and curl, and the ciliated bands become more prominent. By day 32, early brachiolaria are presented with three pairs of brachiolar arms. Advanced brachiolaria with a well-developed brachiolar complex (three pairs of brachia and central adhesive disc) occur 6 weeks after fertilization. In the field, spawning of the sea star was observed in April to May, settlement form larvae and just settlements seem to occur from June to July, and early juveniles occur from August to September. Although we had not described the end of brachiolaria stage, it can be tentatively estimated that the duration of the pelagic stage of A. amurensis is 40 to 50 days.
Zortuk, Mustafa;Gumus, Hasan Onder;Kilinc, Halil Ibrahim;Tuncdemir, Ali Riza
The Journal of Advanced Prosthodontics
/
v.4
no.4
/
pp.192-196
/
2012
PURPOSE. The purpose of this study was to evaluate the effect of provisional cement removal by different dentin cleaning protocols (dental explorer, pumice, cleaning bur, Er:YAG laser) on the shear bond strength between ceramic and dentin. MATERIALS AND METHODS. In total, 36 caries-free unrestored human third molars were selected as tooth specimens. Provisional restorations were fabricated and cemented with eugenol-free provisional cement. Then, disc-shaped ceramic specimens were fabricated and randomly assigned to four groups of dentin cleaning protocols (n = 9). Group 1 (control): Provisional cements were mechanically removed with a dental explorer. Group 2: The dentin surfaces were treated with a cleaning brush with pumice Group 3: The dentin surfaces were treated with a cleaning bur. Group 4: The provisional cements were removed by an Er:YAG laser. Self-adhesive luting cement was used to bond ceramic discs to dentin surfaces. Shear bond strength (SBS) was measured using a universal testing machine at a 0.05 mm/min crosshead speed. The data were analyzed using a Kolmogorov Smirnov, One-way ANOVA and Tukey HSD tests to perform multiple comparisons (${\alpha}$=0.05). RESULTS. The dentin cleaning methods did not significantly affect the SBS of ceramic discs to dentin as follows: dental explorer, pumice, cleaning bur, and Er:YAG laser. CONCLUSION. The use of different cleaning protocols did not affect the SBS between dentin and ceramic surfaces.
Proliposomal patch of clenbuterol, ${\beta}_2-agonist$ bronchodilator, was prepared and its feasibility as a novel transdermal drug delivery system was examined. Proliposomal granules containing clenbuterol was prepared by a standard method using sorbitol and lecithin with (Rx 2) or without cholesterol (Rx 1). The porous structure of sorbitol in the proliposomes was maintained allowing tree flowability of the granules. Following contact with water, the granules were converted probably to liposomes almost completely within several minutes. It indicates that proliposomes may be hydrated, when they are applied on the skin under occlusive condition in vivo, by the sweat to form liposomes. Clenbuterol release from Rx 1 and Rx 2 proliposomes to pH 7.4 isotonic phospate buffer (PBS) across cellulose membrane (mol. wt. cut-off of 12000-14000) was retarded significantly compared with that from the mixture of clenbuterol powder and blank proliposomes. Interestingly, proliposomes prepared with lecithin and cholesterol (i.e., Rx 2 proliposomes) showed much more retarded release of clenbuterol than proliposomes prepared only with lecithin (i.e.. Rx 1 proliposomes), indicating that clenbuterol release from proliposomes can be controlled by the addition of cholesterol to the proliposomes. Proliposomal patches were prepared using PVC film as an occlusive backing sheet, two sides adhesive tape (urethane, 1.45 mm thickness) as a reservoir for proliposome granules and Millipore MF-membrane (0.45 mm pore size) as a drug release-controlling membrane. Rx 1 or Rx 2 proliposomes containing 4.6 mg of clenbuterol were loaded into the reservoir of the patch. Clenbuterol release from the patches to pH 7.4 PBS was determined using USP paddle (50 rpm)-over-disc release method. Clenbuterol release from the proliposomal patches was much more retarded even than from a matrix type clenbuterol patch (Boehringer Ingelheim ltd). Being consistent with clenbuterol release from the proliposomal granules, the release from the patches was highly dependent on the presence of cholesterol in the proliposomes : Patches containing Rx 2 proliposomes showed several fold slower drug release than patches containing Rx 1 proliposomes. When the patch containing Rx 1 proliposomes was applied on to the back of a hair-removed rat, clenbuterol concentration in the rat blood was maintained during 6-72 hrs. Transdermal absorption of clenbuterol from the patch was accelerated when the patch was prehydrated with 50 ml of pH 7.4 PBS before topical application. Above results indicate that sustained transdermal delivery of clenbuterol is feasible using proliposomal patches if the cholesterol content and pore size of the release rate-controlling membrane of patches, for example, are appropriately controlled.
PURPOSE. To evaluate the effects of surface treatments on shear bond strength (SBS) between microwave and conventionally sintered zirconia core/veneers. MATERIALS AND METHODS. 96 disc shaped Noritake Alliance zirconia specimens were fabricated using YenaDent CAM unit and were divided in 2 groups with respect to microwave or conventional methods (n=48/group). Surface roughness (Ra) evaluation was made with a profilometer on randomly selected microwave (n=10) and conventionally sintered (n=10) cores. Specimens were then assessed into 4 subgroups according to surface treatments applied (n=12/group). Groups for microwave (M) and conventionally (C) sintered core specimens were as follows; $M_C$,$C_C$: untreated (control group), $M_1,C_1:Al_2O_3$ sandblasting, $M_2,C_2$:liner, $M_3,C_3:Al_2O_3$ sandblasting followed by liner. Veneer ceramic was fired on zirconia cores and specimens were thermocycled (6000 cycles between $5^{\circ}-55^{\circ}C$). All specimens were subjected to SBS test using a universal testing machine at 0.5 mm/min, failure were evaluated under an optical microscope. Data were statistically analyzed using Shapiro Wilk, Levene, Post-hoc Tukey HSD and Student's t tests, Two-Way-Variance- Analysis and One-Way-Variance-Analysis (${\alpha}$=.05). RESULTS. Conventionally sintered specimens ($1.06{\pm}0.32{\mu}m$) showed rougher surfaces compared to microwave sintered ones ($0.76{\pm}0.32{\mu}m$)(P=.046), however, no correlation was found between SBS and surface roughness (r=-0.109, P=.658). The statistical comparison of the shear bond strengths of $C_3$ and $C_1$ group (P=.015); $C_C$ and $M_C$ group (P=.004) and $C_3$ and $M_3$ group presented statistically higher (P=.005) values. While adhesive failure was not seen in any of the groups, cohesive and combined patterns were seen in all groups. CONCLUSION. Based on the results of this in-vitro study, $Al_2O_{3-}$ sandblasting followed by liner application on conventionally sintered zirconia cores may be preferred to enhance bond strength.
Most investigators recommended that porcelain surface should be roughened with abrasives and/or be etched with acid in repairing the fractured porcelain with composite resin. This study was designed to evaluate the effect of porcelain surface treatments on the bond strength between porcelain and composite resin by measuring the shear bond strength and observing the porcelain surface with SEM. 48 porcelain disc were fabricated with Vintage porcelain and embedded in epoxy resin with the test surface exposed. The specimens were divided four groups at random and the test surfaces of the four groups were prepared as follows : Group 1 : Porcelain surface was roughened with a fine diamond and treated with 32% phosphoric acid gel for 10 seconds. Group 2 : Porcelain surface was roughened with a fine diamond and etched with 8% hydrofluoric acid gel for 5 minutes. Group 3 : Porcelain surface was roughened with a coarse diamond and treated with 32% phosphoric acid gel for 10 seconds. Group 4 : Porcelain surface was roughened with a coarse diamond and etched with 8% hydrofluoric acid gel for 5 minutes. All specimens were washed for 30 seconds. A representative specimen of each group was selected and the porcelain surface was observed with SEM at 1000 magnification. Remaining specimens were silanated, bonded with composite resin, thermocycled, and shear-tested on specially designed zig connected to Instron machine. The results were as follows : 1. The shear bond strength of the group etched with hydrofluoric acid was significantly higher than that of group treated with phosphoric acid(p<0.01). 2. The shear bond strength of the group roughened with a fine diamond was not significantly different from that of the group roughened with a coarse diamond(p>0.01). 3. SEM examination of prepared porcelain surfaces revealed that the surface etched with hydrofluoric acid showed numerous microporosities, undercut, and rougher surface than the surface treated with phosphoric acid. 4. All specimens etched with hydrofluoric acid showed cohesive failure within porcelain, but specimens treated with phosphoric acid mainly showed adhesive failure between porcelain and composite resin.
Ti-AI-Si-N coating layers were deposited on WC-Co substrates by a hybrid system of arc ion plating and sputtering techniques. The coatings were prepared with different Si contents to investigate the effect of Si content on their mechanical properties and microstructures. The dry sliding wear experiments were conducted on Ti-AI-Si-N coated WC-Co discs at constant load, 3N, and sliding speed, 0.1 m/s with two different counterpart materials such as steel ball and zirconia ball using a conventional ball-on-disc sliding wear apparatus. In the case of steel ball, the friction coefficient of Ti-AI-Si-N coating layers became lower than that of Ti-AIN coating layers. The friction coefficient decreased with increasing of Si content due to adhesive wear behavior between coating layer and steel ball. On the contrary, in the case of zirconia ball, the friction coefficient increased with increasing of Si content, indicating that abrasive wear behavior was more dominant when the coating layers slid against zirconia ball.
Lee Seung Hyoun;Suh Su-Jeong;Yun Geum-Hee;Son Yongkeun
Journal of the Korean Electrochemical Society
/
v.7
no.2
/
pp.100-107
/
2004
The nano or micro sized structures of conducting polymer had been prepared by synthesizing the desired polymer within the pores of template of nano or micro porous membrane filter. In this study, we had tried to fabricate conducting polymer microstructures on an electrode by using electrochemical deposition adopting template synthesis. Our attention was focused on two different things, attaching template on the electrode and fabricating microstructures only at limited areas of the electrode. A conducting polymer, PEDiTT (poly 3,4-ethylenedithi-athiophene) solution was blended with PVA(polyvinyl alcohol) solution and used as an conducting adhesive. After attaching template membrane, the electrode were immersed in 0.5M pyrrole in 0.1M KCI solution, and electrochemical polymerization was performed. The growth process of the microstructures studied by SEM. The electrochemical fabrication of conducting polymer was performed by using two-electrode system. A large working electrode and a micro scale disc electrode were used for the confined area synthesis. Polymerization potential was 4V in an electrolytic solution made of KCI in deionized water. The optimum polymerization conditions were, i.e. (4V/100sec) for $250{\mu}m$ electrode and (6V/30 sec) for $10{\mu}m$ electrode.
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