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PubMedDate not supplied

The transposable element IS4712 prevents S-layer gene (sbsA) expression in Bacillus stearothermophilus and also affects the synthesis of altered surface layer proteins.

Cell surface (S)-layer protein synthesis in Bacillus stearothermophilus PV72/p6 is blocked when cells are grown at elevated temperature. From a culture exhibiting the S-layer-negative phenotype, the S-layer deficient mutant T5 (SbsA-) was isolated. Genetic analysis of the S-layer-encoding gene (sbsA) of mutant T5 revealed an insertion element (IS4712) integrated into the upstream regulatory region of the S-layer gene, thereby blocking sbsA transcription. The insertion element consists of 1371 base pairs which are flanked by two perfect inverted terminal repeats. Sequence similarity to other transposases of the IS4 family was detected. DNA-DNA hybridizations demonstrated that multiple homologues of IS4712 were also present within the genomes of several other thermophilic bacillus isolates. Attempts to isolate SbsA+ revertants failed. Instead, cells with altered surface proteins were detected. The synthesis of the altered S-layer proteins was correlated with the presence of IS4712 along with the occurrence of deletions in the sbsA coding region. Furthermore imprecise excision of IS4712 was detected. This work demonstrated that B. stearothermophilus is able to express at least four different S-layer proteins and that blocking of sbsA transcription by the insertion element IS4712 is associated with the expression of altered surface proteins.

5' Untranslated Regions↗

PubMed1989

Three-dimensional structure of the surface protein layer (MW layer) of Bacillus brevis 47.

The three-dimensional (3D) structure of one surface protein layer from Bacillus brevis 47, the middle wall (MW) layer, has been reconstructed from tilted-view electron micrographs after correlation averaging to a resolution of 2 nm. The MW layer has p6 symmetry with a center-to-center spacing of 18.3 nm and a minimum thickness of 5.5 nm. The reconstruction reveals a distinct domain structure: the heavier domain of six monomers jointly forms a massive core centered at the sixfold symmetry axis, and lighter domains interconnect adjacent unit cells. In addition, the larger domains collectively form a pore by making contact with each other towards the inner surface, while the smaller domains establish a second connectivity towards the outer surface of the S layer. The MW layer of B. brevis resembles the S layer of Acetogenium kivui in various aspects: they have very similar lattice parameters and highly reminiscent 3D structures; the pores penetrate through the whole core and appear to determine the porosity of the S layers.

Bacillus↗

PubMed1996

Immunoreactivity of allergen (Bet v 1) conjugated to crystalline bacterial cell surface layers (S-layers).

BACKGROUND: Crystalline cell surface layers (S-layers) from Gram-positive eubacteria had been demonstrated as carrier/adjuvants for chemically synthesized tumor-associated oligosaccharide haptens and capsular polysaccharide antigens of Streptococcus pneumoniae strains. OBJECTIVES: The applicability of S-layers as vaccine carrier for treatment of Type I allergy was investigated. STUDY DESIGN: Native or cross-linked S-layer self-assembly products and cell wall preparations from Bacillus sphaericus CCM 2177 and Thermoanaerobacter thermohydrosulfuricus L111-69 and L110-69 were used for immobilization of recombinant major birch pollen allergen Bet v 1. RESULTS AND CONCLUSIONS: Depending on the carrier used, amounts of approximately 20-40 micrograms allergen per mg conjugate could be immobilized. By application of L-glutamic acid dimethyl ester as a spacer, this value could be increased approximately 10-fold. The functionality of the rBet v 1-conjugates was assessed in immunological systems. (i) The presence of intact B-cell epitopes was demonstrated in inhibition experiments using human Bet v 1-specific IgE. (ii) The rBet v 1-S-layer conjugates were immunogenic in mice. (iii) The proliferation of rBet v 1-specific T-cell clones suggested that the peptides created by processing of immobilized Bet v 1 were similar to those derived from natural allergen. (iv) Stimulation of human allergen-specific TH2 lymphocytes with S-layer-conjugated Bet v 1 led to a modulation of the cytokine production pattern from TH2 to TH0/TH1. This study indicates that S-layers may be suitable carriers for few immunotherapeutical vaccines for Type 1 hypersensitivity.

Adjuvants, Immunologic↗

PubMed2005

Polysaccharide-protein surface modification of titanium via a layer-by-layer technique: characterization and cell behaviour aspects.

To improve the surface biocompatibility of titanium films, a layer-by-layer (LBL) self-assembly technique, based on the polyelectrolyte-mediated electrostatic adsorption of chitosan (Chi) and gelatin (Gel), was used leading to the formation of multilayers on the titanium thin film surfaces. The film growth was initialized by deposition of one layer of positively charged poly(ethylene imine) (PEI). Then the thin film was formed by the alternate deposition of negatively charged Gel and positively charged Chi utilizing electrostatic interactions. The LBL film growth was monitored by several techniques. The chemical composition, surface topography as well as wettability were investigated by using X-ray photoelectron spectroscopy (XPS), atomic force microscopy (AFM), confocal laser scanning microscopy (CLSM) and water contact angle measurement, respectively. Quantitative XPS analysis showed the alternative change of C/N ratio after four sequential cycles coating of Ti/PEI/Gel/Chi/Gel, which indicated the discrete layer structure of coatings. Uncoated titanium (control sample) displayed a smooth surface morphology (root mean square (RMS) roughness was around 2.5 nm). A full coverage of coating with Gel/Chi layers was achieved on the titanium surface only after the deposition layers of PEI/(Gel/Chi)2. The PEI/Gel/(Chi/Gel)3 layer displayed a rough surface morphology with a tree-like structure (RMS roughness is around 82 nm). These results showed that titanium films could be modified with Chi/Gel which may affect the biocompatibility of the modified titanium films. To confirm this hypothesis, cell proliferation and cell viability of osteoblasts on LBL-modified titanium films as well as control samples were investigated in vitro. The proliferation of osteoblasts on modified titanium films was found to be greater than that on control (p<0.05) after 1 and 7 days culture, respectively. Cell viability measurement showed that the Chi/Gel-modified films have higher cell viability (p<0.05) than the control. These data suggest that Chi/Gel were successfully employed to surface engineer titanium via LBL technique, and enhanced its cell biocompatibility. The approach presented here may be exploited for fabrication of titanium-based implant surfaces.

Adsorption↗

PubMed2005

Comparative analysis of cortical layering and supragranular layer enlargement in rodent carnivore and primate species.

The mammalian cerebral cortex is composed of individual layers characterized by the cell types they contain and their afferent and efferent connections. The current study examined the raw, and size-normalized, laminar thicknesses in three cortical regions (somatosensory, motor, and premotor) of fourteen species from three orders of mammals: primates, carnivores, and rodents. The proportional size of the pyramidal cell layers (supra- and infragranular) varied between orders but was similar within orders despite wide variance in absolute cortical thickness. Further, supragranular layer thickness was largest in primates (46 +/- 3 percent), followed by carnivores (36 +/- 3 percent), and then rodents (19 +/- 4 percent), suggesting a distinct difference in the proportion of cortex devoted to corticocortical connectivity across these orders. Although measures of supragranular layer thickness are highly correlated with measures of overall brain size, such associations are not present when independent contrasts are used to control for phylogenetic inertia. Interestingly, neurogenesis time span remains strongly associated with supragranular layer thickness despite size normalization and controlling for phylogenetic inertia. Such layering differences between orders, and similarities amongst species within an order, suggest that supragranular layer expansion may have occurred early in mammalian evolution and may be related to ontogenetic variables such as neurogenesis time span rather than measures of overall size.

Animals↗

PubMed2004

Multi-layer compression: comparison of four different four-layer bandage systems applied to the leg.

OBJECTIVE: To compare performance of four commercial four-layer bandage systems when applied to the leg. METHODS: Four experienced bandagers applied each system: [Profore Regular (Smith and Nephew); Ultra-Four (Robinson); System 4 (Seton) and K Four (Parema)] to the same leg. Bandages were applied as single layers and as completed systems using standard techniques. For each application, 18 pressure measurements were taken using the Borgnis Medical Stocking Tester (MST) at three measuring points (ankle, gaiter and mid-calf) on medial and lateral aspects in three postures: (horizontal, standing and sitting). RESULTS: In all 2304 observations were made, 576 for each bandager, 576 for each bandaging system, 768 for each measuring point, 1152 for each aspect and 768 for each posture. The increase in pressure produced by each additional layer was 65-75% of the pressure of the same bandage when used as a single layer. There were significant differences in the final pressures achieved by the bandagers (means: 45-54 mmHg, p<0.001) and between bandage systems (means: System 4: 46 mmHg, Profore: 47 mmHg, K Four: 52 mmHg, Ultra-Four: 54 mmHg; p=0.005). The relationships between the final pressures achieved at each of the three measuring points, the three postures and the two aspects were not consistent among the bandage systems (p<0.01). CONCLUSIONS: When a bandage is applied as part of a multi-layered system it exerts approximately 70% of the pressure exerted when applied alone, thus challenging the commonly-held assumption that the final pressure achieved by a multi-layer bandaging system is the sum of the pressures exerted by each individual layer. Each of the four bandaging systems exerted different final pressures and gradients and different changes with posture change. These differences have important implications, which could influence the selection (or avoidance) of a particular bandage system according to a patient's condition and circumstances.

Bandages↗

PubMed2007

A factorial, randomized trial of pentoxifylline or placebo, four-layer or single-layer compression, and knitted viscose or hydrocolloid dressings for venous ulcers.

OBJECTIVES: We evaluated the effectiveness of pentoxifylline, knitted viscose or hydrocolloid dressings, and single-layer or four-layer bandaging for venous ulceration. METHOD: A factorial randomized controlled trial with 24-week follow-up was conducted in leg ulcer clinics in Scotland with blinded allocation to pentoxifylline (1200 mg) or placebo, knitted viscose or hydrocolloid dressings, and single-layer or four-layer bandages. The study enrolled 245 adults with venous ulcers. The main outcome measure was time to complete healing. Secondary outcomes included proportions healed, withdrawals, and adverse events. Analysis was by intention to treat. RESULTS: There was no evidence of interaction between the drug, bandages, and dressings. Pentoxifylline was associated with nonsignificant increased ulcer healing (62% vs 53%; P = .21). Four-layer bandages were associated with significantly higher healing rates (67% vs 49%; P = .009). There was no difference in healing between knitted viscose and hydrocolloid dressings (58% and 57%; P = .88). Cox regression models increased the significance of the pentoxifylline effect (relative risk of healing, 1.4; 95% confidence interval, 1.0 to 2.0). CONCLUSIONS: Pentoxifylline increased the proportion healing compared with placebo to the same extent as shown in recent systematic reviews, although this finding was only statistically significant when a secondary adjusted analysis was conducted. Four-layer bandaging produced higher healing rates than single-layer bandaging. There was no difference in time to healing between knitted viscose and hydrocolloid dressings.

Administration, Oral↗

PubMed2000

The effect of hybrid layer thickness on bond strength: demineralized dentin zone of the hybrid layer.

OBJECTIVES: The purpose of this study was to evaluate the correlation between hybrid layer thickness and bond strength using specimens acid-conditioned for varying lengths of time. METHODS: The dentin surfaces of human premolars, sectioned to remove the enamel from the labial surface, were conditioned with 35.0% phosphoric acid of an adhesive resin system (Scotchbond Multi-Purpose; 3M) for 15 (as directed by the manufacturer), 60, 120, or 180 s (experimental acid-conditioning times). The bonded specimens were then sectioned perpendicular to the adhesive interface to measure the hybrid layer thickness by SEM. The specimens for the micro-tensile test were sectioned perpendicular to the adhesive interface and trimmed to an hourglass-shape. Then, the micro-tensile test was performed at a crosshead speed of 1.0 mm/min. The bond strengths and hybrid layer thickness were statistically compared with Student's t-test (p < 0.05). All fractured surfaces were also observed by SEM. RESULTS: Significant differences between the groups exposed to acid for 15 and 60 s, and those exposed for 120 and 180 s were observed in hybrid layer thickness and bond strength (p < 0.05). SEM observation of the fractured surfaces revealed that a demineralized dentin zone without resin impregnation remained within the hybrid layer. SIGNIFICANCE: A demineralized dentin zone was formed in the bond structures after prolonged acid-conditioning, resulting in low bond strength. The shrinkage of the hybrid layer due to desiccation during the SEM examination process provided evidence of the presence of the demineralized dentin zone within the hybrid layer.

Acid Etching, Dental↗

PubMed1999

Temperature elevations computed for three-layer and four-layer obstetrical tissue models in nonlinear and linear ultrasonic propagation cases.

The authors computed temperature elevations in a three-layer and a four-layer tissue model, assuming the crucial obstetrical case when the ultrasonic pulse propagating through the abdominal wall and the fluid-filled bladder penetrates into soft fetal tissues. To consider nonlinear propagation, the authors applied a new theory of nonlinear increase of absorption recently developed by the first author. Computations were carried out for pulses with a carrier frequency of 3 MHz, duration time of 1.33 micros, and pulse repetition frequency of 3.3 kHz. Similar computations were carried out for a four-layer tissue model corresponding to the third trimester of gestation. The ceramic piezoelectric transducer 2 cm in diameter radiated the ultrasonic beam focused at a distance of 6.5 cm. The intensities at the radiating transducer (at the source) were I(SAPA) = 10 and 5 W/cm2. Temperature elevations and distributions were determined numerically for various values of low-amplitude absorption coefficients assumed to be the same as attenuation coefficients. It was shown in the three-layer tissue model that the maximum temperature elevation can be about 50% higher for nonlinear than for linear propagation. The maximum fetal temperature elevation in this case was 2.36 degrees C for nonlinear and 1.84 degrees C for linear propagation. The temperature elevation in the abdominal wall was lower than those temperatures when the attenuation of the abdominal wall was assumed to be a low value of 0.05 Np/cm.MHz (0.45 dB/cm.MHz). However, when it was increased to 0.16 Np/cm.MHz (1.4 dB/cm.MHz), the temperature elevation of the abdominal wall reached 3.2 degrees C and the maximum fetal elevation was 1.65 degrees C. In such cases, the abdominal wall became the principal source of heat production. In this case, the difference between fetal temperature elevations for nonlinear and linear propagation was only about 10%. The results obtained in the four-layer tissue model, in which the uterus tissue also was represented, show that temperature elevations in this case are about 3.6 times lower than in the three-layer tissue model, with comparable attenuation of the abdominal wall. Differences between nonlinear and linear propagation in the four-layer tissue model are negligible. The temperature elevations obtained were proportional to the pulse repetition frequency, without changing temperature distributions in the ultrasonic beam. In this manner, fetal temperature elevations can be reduced by reducing the repetition frequency.

Abdominal Muscles↗

PubMed1998

Layer-by-layer construction of enzyme multilayers on an electrode for the preparation of glucose and lactate sensors: elimination of ascorbate interference by means of an ascorbate oxidase multilayer.

A layer-by-layer structure of enzyme multilayers composed of glucose oxidase (GOx) or lactate oxidase (LOx) and ascorbate oxidase (AOx) was prepared on the surface of a platinum electrode. The amperometric response to glucose or lactate was studied in the presence of ascorbic acid as a possible interference. An alternating and repeated deposition of avidin and the biotin-labeled enzymes resulted in the layer-by-layer structure of GOx/AOx and LOx/AOx multilayers. Optical and gravimetric measurements based on an ultraviolet-visible absorption spectroscopy and a quartz crystal microbalance revealed that the enzyme multilayers thus prepared consist of monomolecular layers of the proteins. The GOx/AOx and LOx/AOx enzyme multilayers were useful to eliminate ascorbic acid interference in the glucose and lactate biosensors, because ascorbic acid can be converted to an electrochemically inert form, dehydroascorbic acid, before being oxidized directly on the Pt electrode. Thus, the GOx/AOx or LOx/AOx multilayer-modified biosensors can be used to determine the normal blood level of glucose (5 mM) and lactate (1 mM) in the presence of a physiological level of ascorbic acid (0.1 mM). The effects of the number of the AOx layers and geometry of the enzyme layers in the multilayer on the performance characteristics of the biosensors are discussed.

Ascorbate Oxidase↗

PubMed2003

A highly crystalline layered silicate with three-dimensionally microporous layers.

Layered silicates with three-dimensional microporosity within the layers have the potential to enable new applications in catalysis, adsorption and ion-exchange. Until now no such materials have been reported. However, here we present the synthesis and structure of AMH-3, a silicate with three-dimensionally microporous layers, obtained in high purity and crystallinity. AMH-3 is composed of silicate layers containing eight-membered rings in all three principal crystal directions, and spaced by strontium cations, sodium cations and water molecules. Because of its three-dimensional pore structure, acid and thermal stability, this layered material could find applications in polymer-silicate composites for membrane applications, for synthesis of combined microporous-mesoporous materials, and for the formation of new zeolites and microporous films. Its existence also opens new possibilities for the synthesis of other layered silicates with multidimensional microporous framework layers.

Cations↗

PubMed2003

On the usefulness of an airborne lidar for O3 layer analysis in the free troposphere and the planetary boundary layer.

Ozone vertical profiling with a lidar is well adapted to the spatial and temporal O3 variability analysis either in the free troposphere, when studying the respective impact of chemical production and dynamical processes, or in the planetary boundary layer (PBL) when characterizing the diurnal evolution of ozone plumes during pollution episodes. Comparisons with other measuring techniques (ozonesonde and aircraft in-situ measurements) demonstrate the lidar ability to characterize narrow layers (< 500 m) with a good accuracy (deltaO3 < 5-10 ppb). Application of airborne or ground-based operation of the CNRS airborne ozone lidar show its ability (i) to observe O3 layering above the PBL during two field experiments held to study air pollution in the Po Valley, Northern Italy, and the city of Marseille, Southern France, (ii) to improve airborne campaign planning (real time information on position of O3 layers) and analysis (three-dimensional perspective for layers detected by in-situ measurements) when chemical characterization of narrow O3 layers in the free troposphere is sought, (iii) to map O3 inhomogeneity down to an horizontal scale of 10-20 km within or above the polluted PBL by airborne measurements. For O3 pollution studies, understanding the origin and the life cycle of O3 layering is the first priority, and in this case the optimum use of the lidar remains the continuous operation of a ground-based instrument.

Air Movements↗

PubMed2003

Uterine rupture, perioperative and perinatal morbidity after single-layer and double-layer closure at cesarean delivery.

OBJECTIVE: This study was undertaken to evaluate the risks and benefits of single-layer uterine closure at cesarean delivery on the index and subsequent pregnancy. STUDY DESIGN: A retrospective study of women delivered of their first live-born infants by primary low transverse cesarean delivery (1989-2001) and their subsequent pregnancy at our institution was performed. RESULTS: Of 768 women studied, 267 had single-layer and 501 had double-layer uterine closures in the index pregnancy. Single-layer closure was associated with slightly decreased blood loss (646 vs 690 mL, P<.01), operative time (46 vs 52 minutes, P<.001), endometritis (13.5% vs 25.5%, P<.001), and postoperative stay (3.5 vs 4.1 days, P<.001). In the second pregnancy, prior single-layer closure was not associated with uterine rupture after a trial of labor (0% vs 1.2%, P=.30), or other maternal or infant morbidities. Prior single-layer closure was associated with increased uterine windows (3.5% vs 0.7%, P=.046) at subsequent cesarean delivery. CONCLUSION: Single-layer uterine closure is associated with decreased infectious morbidity in the index surgery, but not uterine rupture or other adverse outcomes in the subsequent gestation.

Adult↗

PubMed2001

The surface layer (S-layer) glycoprotein of Geobacillus stearothermophilus NRS 2004/3a. Analysis of its glycosylation.

Geobacillus stearothermophilus NRS 2004/3a possesses an oblique surface layer (S-layer) composed of glycoprotein subunits as the outermost component of its cell wall. In addition to the elucidation of the complete S-layer glycan primary structure and the determination of the glycosylation sites, the structural gene sgsE encoding the S-layer protein was isolated by polymerase chain reaction-based techniques. The open reading frame codes for a protein of 903 amino acids, including a leader sequence of 30 amino acids. The mature S-layer protein has a calculated molecular mass of 93,684 Da and an isoelectric point of 6.1. Glycosylation of SgsE was investigated by means of chemical analyses, 600-MHz nuclear magnetic resonance spectroscopy, and matrix-assisted laser desorption ionization-time of flight mass spectrometry. Glycopeptides obtained after Pronase digestion revealed the glycan structure [-->2)-alpha-L-Rhap-(1-->3)-beta-L-Rhap-(1-->2)-alpha-L-Rhap-(1-->](n = 13-18), with a 2-O-methyl group capping the terminal trisaccharide repeating unit at the non-reducing end of the glycan chains. The glycan chains are bound via the disaccharide core -->3)-alpha-l-Rhap-(1-->3)-alpha-L-Rhap-(L--> and the linkage glycose beta-D-Galp in O-glycosidic linkages to the S-layer protein SgsE at positions threonine 620 and serine 794. This S-layer glycoprotein contains novel linkage regions and is the first one among eubacteria whose glycosylation sites have been characterized.

Amino Acid Sequence↗

PubMed1991

The cell envelope of the hyperthermophilic archaebacterium Pyrobaculum organotrphum consists of two regularly arrayed protein layers: three-dimensional structure of the outer layer.

The cell envelope of the hyperthermophilic sulphur-reducing archaebacterium Pyrobaculum organotrophum H10 was found to be composed of two distinct hexagonally arranged crystalline protein arrays. Electron microscopic analysis of freeze-etched cells and isolated envelopes in conjunction with image processing showed that the inner layer (lattice centre-to-centre spacing 27.9 nm) is essentially identical to the protein array of Pyrobaculum islandicum GEO3, a complex, rigid structure implicated in the maintenance of cell shape. The outer layer has clear p6 symmetry and a lattice spacing of 20.6 nm. Its three-dimensional structure was reconstructed from a negative stain tilt series of an intact double-layered envelope using Fourier filtration to separate the desired information from the other lattices present. The outer layer is a unique, porous network of blocklike dimers disposed around six-fold axes, and exhibits minimal asymmetry between its inner and outer faces. It appears to be rather loosely associated with the outer surface of the inner layer. In most H10 envelopes, the inner layer is orientated with one base vector exactly perpendicular to the long axis of the cell, so that the cylindrical portion is composed of a series of parallel cell-girdling hoops of hexameric morphological units. All the other known Pyrobaculum strains were found to have a GEO3-type envelope structure, consisting of a single rigid protein array and a fibrous capsule. Although H10 does not possess a capsule, fibrils appear to be sandwiched between the two protein layers.

Archaea↗

PubMed2007

S-layer anchoring and localization of an S-layer-associated protease in Caulobacter crescentus.

The S-layer of the gram-negative bacterium Caulobacter crescentus is composed of a single protein, RsaA, that is secreted and assembled into a hexagonal crystalline array that covers the organism. Despite the widespread occurrence of comparable bacterial S-layers, little is known about S-layer attachment to cell surfaces, especially for gram-negative organisms. Having preliminary indications that the N terminus of RsaA anchors the monomer to the cell surface, we developed an assay to distinguish direct surface attachment from subunit-subunit interactions where small RsaA fragments are incubated with S-layer-negative cells to assess the ability of the fragments to reattach. In doing so, we found that the RsaA anchoring region lies in the first approximately 225 amino acids and that this RsaA anchoring region requires a smooth lipopolysaccharide species found in the outer membrane. By making mutations at six semirandom sites, we learned that relatively minor perturbations within the first approximately 225 amino acids of RsaA caused loss of anchoring. In other studies, we confirmed that only this N-terminal region has a direct role in S-layer anchoring. As a by-product of the anchoring studies, we discovered that Sap, the C. crescentus S-layer-associated protease, recognized a cleavage site in the truncated RsaA fragments that is not detected by Sap in full-length RsaA. This, in turn, led to the discovery that Sap was an extracellular membrane-bound protease, rather than intracellular, as previously proposed. Moreover, Sap was secreted to the cell surface primarily by the S-layer type I secretion apparatus.

Bacterial Proteins↗

PubMed2002

Caulobacter crescentus synthesizes an S-layer-editing metalloprotease possessing a domain sharing sequence similarity with its paracrystalline S-layer protein.

Strains of Caulobacter crescentus elaborate an S-layer, a two-dimensional protein latticework which covers the cell surface. The S-layer protein (RsaA) is secreted by a type I mechanism (relying on a C-terminal signal) and is unusual among type I secreted proteins because high levels of protein are produced continuously. In efforts to adapt the S-layer for display of foreign peptides and proteins, we noted a proteolytic activity that affected S-layer monomers with foreign inserts. The cleavage was precise, resulting in fragments with an unambiguous N-terminal sequence. We developed an assay to screen for loss of this activity (i.e., presentation of foreign peptides without degradation), using transposon and traditional mutagenesis. A metalloprotease gene designated sap (S-layer-associated protease) was identified which could complement the protease-negative mutants. The N-terminal half of Sap possessed significant similarity to other type I secreted proteases (e.g., alkaline protease of Pseudomonas aeruginosa), including the characteristic RTX repeat sequences, but the C-terminal half which normally includes the type I secretion signal exhibited no such similarity. Instead, there was a region of significant similarity to the N-terminal region of RsaA. We hypothesize that Sap evolved by combining the catalytic portion of a type I secreted protease with an S-layer-like protein, perhaps to associate with nascent S-layer monomers to "scan" for modifications.

Amino Acid Sequence↗

PubMedDate not supplied

Hemocompatibility of layer-by-layer hyaluronic acid/heparin nanostructure coating on stainless steel for cardiovascular stents and its use for drug delivery.

In order to develop drug-eluting cardiovascular stents, stainless steel (SUS316L) sheets were coated with hyaluronic acid (HA) and heparin (HEP), and their in vitro characteristics and drug release pattern were investigated. The surface of stainless steel (SS) was treated with nitric acid and followed by anchoring aminotrimethoxysilane (ATMS), then a nanolayer of HA was covalently immobilized onto the surface. Heparin was then covalently bonded to the HA-immobilized SS substrate. After repeating 1 to 5 cycles, 1 to 5 layers polyelectrolyte complex (PEC) nanobrush of HA/HEP were resulted with the thickness ranging from 280 to 630 nm (measured with ellipsometry). A model drug (sirolimus) was loaded in the HA/HEP layers at a density ranging from 1.02 to 3.12 microg/cm2. The SS-ATMS-HA-HEP substrates were evidenced by X-ray photoelectron spectroscope (XPS), contact angle, and AFM measurement. The effect of this surface modification on the coagulation time of the resulting SS substrates was investigated. The results show that the multilayer HA/HEP stainless steel would exhibit higher anticoagulant activity than pure SS substrates. In addition, the results of the in vitro drug delivery study showed that release of sirolimus from the 5-layer-HA-HEP stainless steel was able to maintain more than 30 days. Thus layer-by-layer HA/HEP PEC can improve the hemocompatibility of SS surface and control the drug released rate by multiple layers of HA/HEP PEC. These results indicate that the multi-layer HA/HEP PEC coated stainless steel would be suitable for drug eluting stents.

Adjuvants, Immunologic↗