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1.
Analyst ; 140(9): 3233-8, 2015 May 07.
Article in English | MEDLINE | ID: mdl-25768651

ABSTRACT

Partially selective gold nanoparticle sensors have the sensitivity and selectivity to discriminate and quantify benzene, toluene, ethylbenzene, p-xylene and naphthalene (BTEXN) at concentrations relevant to the US Environmental Protection Agency. In this paper we demonstrate that gold nanoparticle chemiresistors can do so in the presence of 16 other hydrocarbons and that they did not reduce the discriminating power of the array. A two-level full factorial designed experiment was performed on unary, binary, ternary, quaternary, quinary combinations of BTEXN analytes with and without the possibly interfering hydrocarbons. The nominal component concentration of the mixtures was 100 µg L(-1), equivalent to approximately 100 parts per billion (ppb). Concentrations predicted with the random forests method had an average root mean square error of 10-20% of the component concentrations. This level of accuracy was achieved regardless of whether or not the 16 possibly interfering hydrocarbons were present. This work shows that the sensitivity and selectivity of gold nanoparticles chemiresistor sensors towards BTEXN analytes are not unduly affected by the other hydrocarbons that are expected to be present at a petroleum remediation site.

2.
Opt Express ; 19(18): 17167-72, 2011 Aug 29.
Article in English | MEDLINE | ID: mdl-21935079

ABSTRACT

We have demonstrated the nonlinear absorption at 532 nm wavelength in an Au semi-continuous film (SF) resulting from smearing of the Fermi distribution and diffusion of conduction electrons into the substrate. The Au SF was irradiated by a pulsed laser with 8 ns pulse width at 532 nm in near resonance with the interband transition of the Au. We determined the temperature increase in the SF for different intensities by electrical measurement. We calculated the temperature increase by using a 1D heat transport equation; comparing the results of the calculation with measured values for the temperature increase, revealed the nonlinear absorption in the Au SF. We employed this deviation from linear behaviour to determine the nonlinear absorption coefficient.

3.
J Clin Dent ; 22(3): 62-7, 2011.
Article in English | MEDLINE | ID: mdl-21905399

ABSTRACT

OBJECTIVE: The aim of this study was to characterize, in vitro, the mode of action of calcium sodium phosphosilicate (NovaMin) in occluding dentin tubules for the purpose of treating dentin hypersensitivity. METHODS: Calcium sodium phosphosilicate (CSPS) was combined with artificial saliva on surfaces of prepared dentin discs. The layer formed was initially examined by a scanning electron microscope (SEM). Focused ion beam (FIB) milling was used to make bulk cross-sections and thin film lamellae. Low kV scanning transmission electron microscopy (STEM), energy dispersive x-ray spectroscopy (EDS), and selected area electron diffraction were then used to characterize, chemically and structurally, the layer formed and the material occluding the tubules. Experiments were also performed to assess the suitability of using an environmental scanning electron microscope (ESEM) in wet mode to follow the transition from CSPS to hydroxyapatite. RESULTS: SEM imaging showed that a layer was formed on the treated dentin samples, and that this layer occluded tubules. Chemical and structural analysis of this material showed that it was hydroxyapatite-like. The wet mode ESEM experiments demonstrated that this technique has the potential to follow the transition from CSPS to the crystalline hydroxyapatite material. CONCLUSION: The use of modern imaging and analysis techniques has demonstrated, in vitro, the reaction of CSPS from an amorphous material to a crystalline hydroxyapatite-like material. These experiments confirmed an occlusion mode of action for CSPS for the treatment of dentin hypersensitivity.


Subject(s)
Dentin Desensitizing Agents/pharmacology , Dentin Sensitivity/drug therapy , Dentin/drug effects , Glass , Acid Etching, Dental/methods , Calcium Phosphates/chemistry , Calcium Phosphates/pharmacology , Citric Acid/chemistry , Crystallography , Dentin/chemistry , Dentin/ultrastructure , Dentin Desensitizing Agents/chemistry , Durapatite/chemistry , Glass/chemistry , Humans , Materials Testing , Microscopy, Electron, Scanning , Microscopy, Electron, Scanning Transmission , Particle Size , Saliva, Artificial/chemistry , Silicates/chemistry , Silicates/pharmacology , Spectrometry, X-Ray Emission , Time Factors
4.
J Chem Phys ; 129(4): 044708, 2008 Jul 28.
Article in English | MEDLINE | ID: mdl-18681671

ABSTRACT

The room temperature thermoelectric properties of a three-dimensional array of molecular junctions are calculated. The array is composed of n-doped silicon nanoparticles where the surfaces are partially covered with polar molecules and the nanoparticles are bridged by trans-polyacetylene molecules. The role of the polar molecules is to reduce the band bending in the n-doped silicon nanoparticles and to shift the electronic resonances of the bridging molecules to the nanoparticle conduction band edges where the molecular resonances act as electron energy filters. The transmission coefficients of the bridging molecules that appear in the formulas for the Seebeck coefficient, the electrical conductance, and the electronic thermal conductance, are calculated using the nonequilibrium Green's function technique. A simple tight-binding Hamiltonian is used to describe the bridging molecules, and the self-energy term is calculated using the parabolic conduction band approximation. The dependencies of the thermoelectric properties of the molecular junctions on the silicon doping concentration and on the molecule-nanoparticle coupling are discussed. The maximal achievable thermoelectric figure of merit ZT of the array is estimated as a function of the phononic thermal conductance of the bridging molecules and the doping of the nanoparticles. The power factor of the array is also calculated. For sufficiently small phononic thermal conductances of the bridging molecules, very high ZT values are predicted.

5.
Phys Rev Lett ; 95(8): 086403, 2005 Aug 19.
Article in English | MEDLINE | ID: mdl-16196877

ABSTRACT

We report on a field-induced change of the electronic band structure of CeBiPt as evidenced by electrical-transport measurements in pulsed magnetic fields. Above approximately 25 T, the charge-carrier concentration increases nearly 30% with a concomitant disappearance of the Shubnikov-de Haas signal. These features are intimately related to the Ce 4f electrons since for the non-4f compound LaBiPt the Fermi surface remains unaffected. Electronic band-structure calculations point to a 4f-polarization-induced change of the Fermi-surface topology.

8.
Phys Rev Lett ; 90(1): 017202, 2003 Jan 10.
Article in English | MEDLINE | ID: mdl-12570643

ABSTRACT

We report the magnetotransport characteristics of a trilayer ferromagnetic tunnel junction built of an electron doped manganite (La0.7Ce0.3MnO3) and a hole doped manganite (La0.7Ca0.3MnO3). At low temperatures the junction exhibits a large positive tunneling magnetoresistance (TMR), irrespective of the bias voltage. At intermediate temperatures below T(C) the sign of the TMR is dependent on the bias voltage across the junction. The magnetoresistive characteristics of the junction strongly suggest that La0.7Ce0.3MnO3 is a minority spin carrier ferromagnet with a high degree of spin polarization, i.e., a transport half-metal.

9.
EMBO J ; 17(10): 2846-54, 1998 May 15.
Article in English | MEDLINE | ID: mdl-9582278

ABSTRACT

We have shown previously that the heavy metal-responsive transcriptional activator MTF-1 regulates the basal and heavy metal-induced expression of metallothioneins. To investigate the physiological function of MTF-1, we generated null mutant mice by targeted gene disruption. Embryos lacking MTF-1 die in utero at approximately day 14 of gestation. They show impaired development of hepatocytes and, at later stages, liver decay and generalized edema. MTF-1(-/-) embryos fail to transcribe metallothionein I and II genes, and also show diminished transcripts of the gene which encodes the heavy-chain subunit of the gamma-glutamylcysteine synthetase, a key enzyme for glutathione biosynthesis. Metallothionein and glutathione are involved in heavy metal homeostasis and detoxification processes, such as scavenging reactive oxygen intermediates. Accordingly, primary mouse embryo fibroblasts lacking MTF-1 show increased susceptibility to the cytotoxic effects of cadmium or hydrogen peroxide. Thus, MTF-1 may help to control metal homeostasis and probably cellular redox state, especially during liver development. We also note that the MTF-1 null mutant phenotype bears some similarity to those of two other regulators of cellular stress response, namely c-Jun and NF-kappaB (p65/RelA).


Subject(s)
Edema/etiology , Liver Diseases/etiology , Liver/embryology , Transcription Factors/physiology , Animals , Cadmium/pharmacology , Cells, Cultured , DNA-Binding Proteins , Dipeptides , Embryonic and Fetal Development , Female , Fetal Diseases , Fibroblasts , Gene Expression Regulation , Gene Targeting , Glutathione Synthase/genetics , Hydrogen Peroxide/pharmacology , Liver/pathology , Male , Mice , Mice, Inbred C57BL , Mice, Knockout , Phenotype , Transcription Factors/genetics , Transcription Factor MTF-1
10.
J Bacteriol ; 179(4): 1082-9, 1997 Feb.
Article in English | MEDLINE | ID: mdl-9023187

ABSTRACT

The major surface protein (Msp) of Treponema denticola has been implicated as a mediator of the interaction between the spirochete and the gingival epithelium in periodontal diseases. Previous studies showed that the Msp of T. denticola ATCC 35405 had porin activity, depolarized epithelial cell membranes, bound to extracellular matrix components of epithelial cells, and formed a regular hexagonal surface array in the treponemal outer membrane. The gene encoding Msp in ATCC 35405 was recently cloned, sequenced, and expressed in Escherichia coli (J. C. Fenno, K.-H. Müller, and B. C. McBride, J. Bacteriol. 178:2489-2496, 1996). In the present study, we identified genes encoding Msp-like proteins in several oral spirochetes. A prominent heat-modifiable Msp-like protein having an apparent molecular mass of between 43 and 64 kDa was present in all oral spirochete strains tested. Antibodies raised against the ATCC 35405 Msp reacted strongly with the Msp proteins of T. denticola ATCC 35404 and T. vincentii, reacted very weakly with the Msp protein of T. denticola ATCC 33520, and did not react with T. denticola OTK, T. socranskii, and T. pectinovorum. The msp loci of the T. denticola strains and T. vincentii were identified in analyses using PCR with oligonucleotide primers derived from the DNA sequence flanking msp in ATCC 35405. Southern blot analysis showed at least three groups of related msp DNA sequences. Comparison of DNA sequences of the 5' and 3' ends of the msp genes showed high sequence homology in the flanking regions and signal peptide coding regions, while the homologies between regions encoding the mature peptide were as low as 50%. The entire msp DNA sequences of T. denticola ATCC 33520 and OTK were determined, and the deduced Msp amino acid sequences were compared to the sequence of the previously reported Msp of ATCC 35405. The results show that the msp locus is conserved in oral treponemes but that there are significant differences between the mature Msp peptides of different strains. Further studies of the antigenic domains, functional domains, and physical structures of Msp proteins, based on these results, will enhance understanding of the role of Msp in the cytopathology associated with oral spirochetes.


Subject(s)
Bacterial Proteins , Conserved Sequence , Genes, Bacterial , Porins/genetics , Treponema/genetics , Amino Acid Sequence , Base Sequence , Blotting, Southern , Cell Membrane/ultrastructure , Molecular Sequence Data , Polymerase Chain Reaction , Porins/analysis , Porins/chemistry , Protein Structure, Secondary , Sequence Alignment , Sequence Homology, Nucleic Acid , Treponema/chemistry , Treponema/ultrastructure
11.
Unfallchirurg ; 99(8): 561-8, 1996 Aug.
Article in German | MEDLINE | ID: mdl-8975377

ABSTRACT

Between 1990 and 1994, 94 patients over 65 years with subcapital humerus fractures were treated by plate osteosynthesis. All patients were operated on within 48 h after trauma. Intensive physical exercising was begun in the early postoperative period. Three months after the operation all osteosynthesis materials were removed with differentiated treatment of the subacromial area. If deemed necessary on the basis of the intraoperative findings (i.e. symptoms of impingement) treatment included a Neer acromioplasty. Sixty-nine of the 94 patients were followed up, including clinical, radiological and ultrasonographic examinations. Twenty-two patients showed an excellent, 26 a good, 9 a satisfactory and 12 an unsatisfactory result, i.e. in more than 82% of patients functional result was excellent to satisfactory. Therefore, we recommended plate osteosynthesis of subcapital humerus fractures in the elderly, in combination with our standard postoperative regimen.


Subject(s)
Bone Plates , Fracture Fixation, Internal , Shoulder Fractures/surgery , Aged , Aged, 80 and over , Female , Humans , Male , Physical Therapy Modalities , Postoperative Complications/diagnostic imaging , Postoperative Complications/rehabilitation , Radiography , Shoulder Fractures/diagnostic imaging , Shoulder Impingement Syndrome/diagnostic imaging , Shoulder Impingement Syndrome/rehabilitation , Treatment Outcome
12.
J Clin Invest ; 98(2): 482-9, 1996 Jul 15.
Article in English | MEDLINE | ID: mdl-8755660

ABSTRACT

Angiogenesis is important in the pathophysiology of endometriosis, a condition characterized by implantation of ectopic endometrium in the peritoneal cavity. Vascular endothelial growth factor (VEGF) is a potent angiogenic factor involved in physiological and pathological angiogenesis, and elevated levels of VEGF are found in peritoneal fluid of patients with endometriosis. Our aim was to investigate the site of expression and regulation of VEGF in endometriosis. VEGF immunoreactivity was found in tissue macrophages present in ectopic endometrium and in activated peritoneal fluid macrophages. Macrophage activation was highest in women with endometriosis, and media conditioned by peritoneal fluid macrophages from these women caused a VEGF-dependent increase in endothelial cell proliferation above that seen from normal women. Peritoneal fluid macrophages secreted VEGF in response to ovarian steroids, and this secretion was enhanced after activation with lipopolysaccharide. Peritoneal fluid macrophages expressed receptors for steroid hormones. VEGF receptors flt and KDR (kinase domain receptor) were also detected, suggesting autocrine regulation. During the menstrual cycle, expression of flt was constant but that of KDR was increased in the luteal phase, at which time the cells migrated in response to VEGF. KDR expression and the migratory response were significantly higher in patients with endometriosis. This study demonstrates that activated macrophages are a major source of VEGF in endometriosis and that this expression is regulated directly by ovarian steroids.


Subject(s)
Endometriosis/physiopathology , Endothelial Growth Factors/biosynthesis , Endothelium, Vascular/cytology , Estradiol/pharmacology , Lymphokines/biosynthesis , Macrophages, Peritoneal/physiology , Progesterone/pharmacology , Adult , Base Sequence , Biological Assay , Cells, Cultured , Culture Media, Conditioned , DNA Primers , Endometriosis/immunology , Endothelial Growth Factors/analysis , Endothelial Growth Factors/pharmacology , Endothelium, Vascular/drug effects , Female , Flow Cytometry , Humans , Lipopolysaccharides/pharmacology , Lymphokines/analysis , Lymphokines/pharmacology , Macrophage Activation , Macrophages, Peritoneal/drug effects , Molecular Sequence Data , Peritoneal Cavity , Polymerase Chain Reaction , Receptor Protein-Tyrosine Kinases/analysis , Receptor Protein-Tyrosine Kinases/biosynthesis , Receptors, Estrogen/biosynthesis , Receptors, Growth Factor/analysis , Receptors, Growth Factor/biosynthesis , Receptors, Progesterone/biosynthesis , Receptors, Vascular Endothelial Growth Factor , Reference Values , Umbilical Veins , Vascular Endothelial Growth Factor A , Vascular Endothelial Growth Factors
13.
J Bacteriol ; 178(9): 2489-97, 1996 May.
Article in English | MEDLINE | ID: mdl-8626313

ABSTRACT

The gene encoding the major outer sheath protein (Msp) of the oral spirochete Treponema denticola ATCC 35405 was cloned, sequenced, and expressed in Escherichia coli. Preliminary sequence analysis showed that the 5' end of the msp gene was not present on the 5.5-kb cloned fragment described in a recent study (M. Haapasalo, K. H. Müller, V. J. Uitto, W. K. Leung, and B. C. McBride, Infect. Immun. 60:2058-2065,1992). The 5' end of msp was obtained by PCR amplification from a T. denticola genomic library, and an open reading frame of 1,629 bp was identified as the coding region for Msp by combining overlapping sequences. The deduced peptide consisted of 543 amino acids and had a molecular mass of 58,233 Da. The peptide had a typical prokaryotic signal sequence with a potential cleavage site for signal peptidase 1. Northern (RNA) blot analysis showing the msp transcript to be approximately 1.7 kb was consistent with the identification of a promoter consensus sequence located optimally upstream of msp and a transcription termination signal found downstream of the stop codon. The entire msp sequence was amplified from T. denticola genomic DNA and cloned in E. coli by using a tightly regulated T7 RNA polymerase vector system. Expression of Msp was toxic to E. coli when the entire msp gene was present. High levels of Msp were produced as inclusion bodies when the putative signal peptide sequence was deleted and replaced by a vector-encoded T7 peptide sequence. Recombinant Msp purified to homogeneity from a clone containing the full-length msp gene adhered to immobilized laminin and fibronectin but not to bovine serum albumin. Attachment of recombinant Msp was decreased in the presence of soluble substrate. Attachment of T. denticola to immobilized laminin and fibronectin was increased by pretreatment of the substrate with recombinant Msp. These studies lend further support to the hypothesis that Msp mediates the extracellular matrix binding activity of T. denticola.


Subject(s)
Bacterial Outer Membrane Proteins/genetics , Bacterial Outer Membrane Proteins/metabolism , Bacterial Proteins , Porins/genetics , Porins/metabolism , Treponema/genetics , Amino Acid Sequence , Base Sequence , Cloning, Molecular , DNA-Directed RNA Polymerases/genetics , Escherichia coli/genetics , Fibronectins/metabolism , Gene Expression , Genes, Bacterial/genetics , Laminin/metabolism , Molecular Sequence Data , Molecular Weight , Porins/chemistry , Porins/isolation & purification , Promoter Regions, Genetic/genetics , Protein Binding , RNA, Bacterial/analysis , RNA, Messenger/analysis , Recombinant Fusion Proteins , Sequence Analysis, DNA , Viral Proteins
14.
Article in German | MEDLINE | ID: mdl-9102022

ABSTRACT

Between 1990 and 1994 94 patients over 65 years of age with subcapital humeral fractures were treated by plate osteosynthesis combined with a modified postoperative treatment including intensive physical exercise in the early postoperative period. Three months after surgery all osteosynthesis materials were removed using a differentiated treatment of the subacromial area. In the follow-up study 69 of the 94 patients were examined; 22 patients showed an excellent, 26 a good, 9 a satisfactory, and 12 an unsatisfactory result. Thus, we recommend plate osteosynthesis of subcapital humeral fractures in elderly people in combination with our standardized postoperative regimes.


Subject(s)
Bone Plates , Fracture Fixation, Internal/methods , Shoulder Dislocation/surgery , Shoulder Fractures/surgery , Aged , Combined Modality Therapy , Follow-Up Studies , Fracture Healing/physiology , Humans , Physical Therapy Modalities , Reoperation , Treatment Outcome
15.
Anal Bioanal Chem ; 353(3-4): 369-71, 1995 Oct.
Article in English | MEDLINE | ID: mdl-15048501

ABSTRACT

The recently developed High Frequency Mode HFM of electron gas SNMS allows investigations on insulating samples with the well known advantages of the SNMS Direct Bombardment Mode DBM for the analysis of conducting samples. HFM has been applied to analyses of different historic ceramic and glass samples in order to demonstrate the possibilities of SNMS in this field. It is shown that manufacturing places of ceramic samples could be distinguished by SNMS mass spectra ("fingerprints"). Furthermore questions of the constituents of colour remains on a painted ceramic ("Kaisermedaillon") could be answered by our SNMS analyses. SNMS investigations have been also applied to corrosion phenomena on different glass samples.

16.
Somat Cell Mol Genet ; 21(5): 289-97, 1995 Sep.
Article in English | MEDLINE | ID: mdl-8619126

ABSTRACT

Heavy metal-induced transcription in mammalian cells is conferred by the metal-responsive 70 kDa transcription factor MTF-1 which contains six zinc fingers and at least three activation domains. In previous cell transfection experiments we have shown that the zinc finger region confers an about 3 fold metal inducibility of transcription, due to its differential zinc binding. However, we also noted that human MTF-1 was more metal-responsive than the mouse factor (about 10 fold versus 3 fold, respectively). Here we analyze this difference in more detail by using chimeric human-mouse factors and narrow the critical region to a 64 amino acid stretch immediately downstream of the zinc fingers, overlapping with the acidic activation domain. A short human segment of this region (aa 313-377) confers efficient metal induction to the mouse MTF-1 when replacing the corresponding mouse region. However, high metal inducibility requires an unaltered MTF-1 and is lost when human MTF-1 is fused to the general activation domain of herpesvirus VP16. Wild type and truncation mutants of MTF-1 fused to VP16 yield chimeras of high transcriptional activity, some exceeding the wildtype regulator, but only limited (about 3 fold) heavy metal inducibility.


Subject(s)
Transcription Factors/metabolism , 3T3 Cells , Amino Acid Sequence , Animals , Base Sequence , DNA-Binding Proteins , Gene Expression/drug effects , HeLa Cells , Herpes Simplex Virus Protein Vmw65/biosynthesis , Herpes Simplex Virus Protein Vmw65/metabolism , Humans , Mammals , Metals/pharmacology , Mice , Molecular Sequence Data , Recombinant Fusion Proteins/biosynthesis , Recombinant Fusion Proteins/metabolism , Restriction Mapping , Sequence Homology, Amino Acid , Sequence Homology, Nucleic Acid , Transcription Factors/biosynthesis , Transcription Factors/genetics , Transcription, Genetic/drug effects , Transcriptional Activation , Zinc Fingers , Transcription Factor MTF-1
17.
Aktuelle Traumatol ; 24(1): 38-41, 1994 Feb.
Article in German | MEDLINE | ID: mdl-8165959

ABSTRACT

The great increase in the number of persons engaged in sports in the 30-50 years age group has led to a marked increase in injuries of the tendon of Achilles. In this case there is tearing loose of the Achilles tendon; the bone and the tendon come apart, as in a rupture of Sharpey's fibres. The tack was made in webbing the tendon of Achilles with polydioxanone ligaments to the tuberosity of calcaneus. The question about prophylactic webbing of the degenerated tendon of Achilles on the other side has to be discussed. Assessment of this injury from the viewpoint of the accident insurance company is explained.


Subject(s)
Achilles Tendon/injuries , Athletic Injuries/surgery , Tendon Injuries/surgery , Tennis/injuries , Achilles Tendon/diagnostic imaging , Achilles Tendon/surgery , Adult , Athletic Injuries/diagnostic imaging , Humans , Male , Postoperative Complications/diagnostic imaging , Rupture, Spontaneous , Suture Techniques , Tendon Injuries/diagnostic imaging , Ultrasonography
18.
J Bacteriol ; 175(9): 2523-33, 1993 May.
Article in English | MEDLINE | ID: mdl-8097515

ABSTRACT

Salmonella enteritidis produces thin, filamentous fimbriae designated SEF14. A 3.9-kb region of a 5.3-kb fragment encoding genes responsible for SEF14 biosynthesis was sequenced and found to contain three genes, sefABC. sefA encoded a novel fimbrin, the structural subunit of SEF14 fimbriae. sefB and sefC encoded proteins homologous to Escherichia coli and Klebsiella pneumoniae fimbrial periplasmic chaperone proteins and fimbrial outer membrane proteins, respectively, and are the first such genes to be characterized from Salmonella spp. in vitro expression directed by the 5.3-kb DNA fragment identified SefA, SefB, and SefC as approximately 14,000-, 28,000-, and 90,000-M(r) proteins, respectively, which correlated with their predicted amino acid sequences. sefB and sefC were not expressed in the absence of sefA. Primer extension analysis of sefABC revealed two major transcription start sites located upstream of sefA. Transcription of sefBC also initiated from the sefA promoter region. Secondary-structure analysis of the mRNA transcript for sefABC predicted the formation of two stable stem-loop structures in the intercistronic region between sefA and sefB indicative of differential regulation of SefA, SefB, and SefC translation. E. coli cells carrying the 5.3-kb DNA fragment of S. enteritidis DNA were unable to assemble distinguishable SEF14 fimbriae; however, immunogold-labelled SEF14 fimbriae were displayed on E. coli clones containing a 44-kb DNA fragment which encompassed the 5.3-kb region. Therefore, sefABC genes make up part of a complex sef operon responsible for the expression and assembly of SEF14 fimbriae.


Subject(s)
Fimbriae Proteins , Fimbriae, Bacterial , Genes, Bacterial/genetics , Membrane Glycoproteins/genetics , Microfilament Proteins , Molecular Chaperones , Proteins/genetics , Salmonella enteritidis/genetics , Amino Acid Sequence , Bacterial Proteins/biosynthesis , Bacterial Proteins/genetics , Base Sequence , Chaperonins , Membrane Glycoproteins/isolation & purification , Microscopy, Immunoelectron , Molecular Sequence Data , Nucleic Acid Conformation , Operon/genetics , RNA, Messenger/genetics , Salmonella enteritidis/ultrastructure , Sequence Analysis, DNA , Sequence Homology, Amino Acid
19.
Infect Immun ; 61(5): 1694-9, 1993 May.
Article in English | MEDLINE | ID: mdl-7682993

ABSTRACT

A 53-kDa protein from the outer sheath of the oral spirochete Treponema denticola was purified to homogeneity and shown to reconstitute channels in black lipid bilayer model membranes. The channel had a single-channel conductance of 1.8 nS in 0.1 M KCl, making this the largest porin channel observed to date (estimated diameter, 3.4 nm). Electron micrographs of 53-kDa-protein-containing outer sheaths of T. denticola showed a regular hexagonal array of darker staining pits.


Subject(s)
Antigens, Bacterial/chemistry , Antigens, Surface/chemistry , Bacterial Outer Membrane Proteins/chemistry , Ion Channels/chemistry , Treponema/chemistry , Bacterial Adhesion , Electric Conductivity , Ion Channel Gating , Membranes, Artificial , Microscopy, Electron , Molecular Weight , Porins , Treponema/ultrastructure
20.
Infect Immun ; 60(5): 2058-65, 1992 May.
Article in English | MEDLINE | ID: mdl-1563796

ABSTRACT

Treponema denticola surface proteins were studied for their biochemical and biological characteristics. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis of detergent extracts of whole cells revealed a major protein of 53 kDa and a number of minor proteins. Antiserum raised against whole cells of T. denticola ATCC 35405 reacted with the 53-kDa protein and a 72-kDa protein but not with the other proteins. Immunoelectron microscopy with anti-53-kDa-protein antibodies showed that the 53-kDa protein is located on the surface of the cell. SDS-PAGE analysis of unheated samples indicated that the 53-kDa protein is the major component of oligomers with molecular masses ranging from 130 to 300 kDa. Western blot (immunoblot) analysis showed that the high-molecular-mass oligomers reacted with whole-cell antiserum and anti-53-kDa-protein antibody. The aggregates dissociated into their subunits after heating to 70 degrees C. Isoelectric focusing followed by SDS-PAGE indicated that the 53-kDa protein was separated into several forms with apparent pI values ranging from 8.0 to 5.5. The oligomeric forms were highly resistant to proteolysis by trypsin and proteinase K, whereas the monomeric proteins were readily digested. A clone expressing a 53-kDa antigen in Escherichia coli was isolated from a lambda ZAP II DNA library of T. denticola ATCC 35405. The recombinant protein had exactly the same molecular mass as the major 53-kDa T. denticola surface protein and reacted with antisera raised against this protein. The role of T. denticola ATCC 35405 surface proteins in attachment to laminin, fibronectin, gelatin, fibrinogen, and bovine serum albumin (BSA) was studied by a modified Western blot binding assay. Fibronectin, laminin, and fibrinogen attached to the 53-kDa surface protein of T. denticola as well as to a 72-kDa protein, whereas no attachment to gelatin or BSA was observed. Attachment could be inhibited by pretreating the blots with fibrinogen but not with gelatin or BSA. Our results suggest that the 53-kDa major surface protein of T. denticola may play a role in the attachment to host proteins and may thus be an important virulence determinant of this species.


Subject(s)
Antigens, Bacterial/analysis , Cloning, Molecular , Treponema/immunology , Animals , Antigens, Bacterial/genetics , Antigens, Bacterial/metabolism , Antigens, Surface/analysis , Antigens, Surface/genetics , Bacterial Adhesion , Electrophoresis, Polyacrylamide Gel , Fibronectins/metabolism , Laminin/metabolism , Molecular Weight , Rabbits , Recombinant Proteins/analysis , Treponema/genetics , Treponema/physiology
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