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Article ; Online: Periodontal bacteria and cardiovascular problems.

Asikainen, Sirkka E

Future microbiology

2009 Volume 4, Issue 5, Page(s) 495–498

MeSH term(s)	Bacteria/immunology ; Bacteria/pathogenicity ; Cardiovascular Diseases/epidemiology ; Cardiovascular Diseases/etiology ; Humans ; Periodontitis/complications ; Periodontitis/microbiology
Language	English
Publishing date	2009-06
Publishing country	England
Document type	Editorial
ISSN	1746-0921
ISSN (online)	1746-0921
DOI	10.2217/fmb.09.21
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Article ; Online: Coaggregation and biofilm growth of Granulicatella spp. with Fusobacterium nucleatum and Aggregatibacter actinomycetemcomitans.

Karched, Maribasappa / Bhardwaj, Radhika G / Asikainen, Sirkka E

BMC microbiology

2015 Volume 15, Page(s) 114

Abstract: Background: Members of fastidious Granulicatella and Aggregatibacter genera belong to normal oral flora bacteria that can cause serious infections, such as infective endocarditis. Aggregatibacter actinomycetemcomitans has long been implicated in ... ...

Abstract	Background: Members of fastidious Granulicatella and Aggregatibacter genera belong to normal oral flora bacteria that can cause serious infections, such as infective endocarditis. Aggregatibacter actinomycetemcomitans has long been implicated in aggressive periodontitis, whereas DNA-based methods only recently showed an association between Granulicatella spp. and dental diseases. As bacterial coaggregation is a key phenomenon in the development of oral and nonoral multispecies bacterial communities it would be of interest knowing coaggregation pattern of Granulicatella species with A. actinomycetemcomitans in comparison with the multipotent coaggregator Fusobacterium nucleatum. The aim was to investigate coaggregation and biofilm formation of Granulicatella elegans and Granulicatella adiacens with A. actinomycetemcomitans and F. nucleatum strains. Results: F. nucleatum exhibited significantly (p < 0.05) higher autoaggregation than all other test species, followed by A. actinomycetemcomitans SA269 and G. elegans. A. actinomycetemcomitans CU1060 and G. adiacens did not autoaggregate. G. elegans with F. nucleatum exhibited significantly (p < 0.05) higher coaggregation than most others, but failed to grow as biofilm together or separately. With F. nucleatum as partner, A. actinomycetemcomitans strains SA269, a rough-colony wild-type strain, and CU1060, a spontaneous smooth-colony laboratory variant, and G. adiacens were the next in coaggregation efficiency. These dual species combinations also were able to grow as biofilms. While both G. elegans and G. adiacens coaggregated with A. actinomycetemcomitans strain SA269, but not with CU1060, they grew as biofilms with both A. actinomycetemcomitans strains. Conclusions: G. elegans failed to form biofilm with F. nucleatum despite the strongest coaggregation with it. The ability of Granulicatella spp. to coaggregate and/or form biofilms with F. nucleatum and A. actinomycetemcomitans strains suggests that Granulicatella spp. have the potential to integrate into dental plaque biofilms.
MeSH term(s)	Aggregatibacter actinomycetemcomitans/physiology ; Bacterial Adhesion ; Biofilms/growth & development ; Carnobacteriaceae/physiology ; Dental Plaque/microbiology ; Fusobacterium nucleatum/physiology ; Humans ; Species Specificity
Language	English
Publishing date	2015-05-30
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2041505-9
ISSN	1471-2180 ; 1471-2180
ISSN (online)	1471-2180
ISSN	1471-2180
DOI	10.1186/s12866-015-0439-z
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Article ; Online: Identification of the pangenome and its components in 14 distinct Aggregatibacter actinomycetemcomitans strains by comparative genomic analysis.

Kittichotirat, Weerayuth / Bumgarner, Roger E / Asikainen, Sirkka / Chen, Casey

PloS one

2011 Volume 6, Issue 7, Page(s) e22420

Abstract: ... major groups were identified; serotypes a, d, e, and f, and serotypes b and c. A serotype e strain was ...

Abstract	Background: Aggregatibacter actinomycetemcomitans is genetically heterogeneous and comprises distinct clonal lineages that may have different virulence potentials. However, limited information of the strain-to-strain genomic variations is available. Methodology/principal findings: The genome sequences of 11 A. actinomycetemcomitans strains (serotypes a-f) were generated de novo, annotated and combined with three previously sequenced genomes (serotypes a-c) for comparative genomic analysis. Two major groups were identified; serotypes a, d, e, and f, and serotypes b and c. A serotype e strain was found to be distinct from both groups. The size of the pangenome was 3,301 genes, which included 2,034 core genes and 1,267 flexible genes. The number of core genes is estimated to stabilize at 2,060, while the size of the pangenome is estimated to increase by 16 genes with every additional strain sequenced in the future. Within each strain 16.7-29.4% of the genome belonged to the flexible gene pool. Between any two strains 0.4-19.5% of the genomes were different. The genomic differences were occasionally greater for strains of the same serotypes than strains of different serotypes. Furthermore, 171 genomic islands were identified. Cumulatively, 777 strain-specific genes were found on these islands and represented 61% of the flexible gene pool. Conclusions/significance: Substantial genomic differences were detected among A. actinomycetemcomitans strains. Genomic islands account for more than half of the flexible genes. The phenotype and virulence of A. actinomycetemcomitans may not be defined by any single strain. Moreover, the genomic variation within each clonal lineage of A. actinomycetemcomitans (as defined by serotype grouping) may be greater than between clonal lineages. The large genomic data set in this study will be useful to further examine the molecular basis of variable virulence among A. actinomycetemcomitans strains.
MeSH term(s)	Actinobacillus/genetics ; Actinobacillus/pathogenicity ; Cluster Analysis ; Comparative Genomic Hybridization/methods ; Gene Pool ; Genes, Bacterial/genetics ; Genome, Bacterial/genetics ; Genomic Islands/genetics ; Molecular Sequence Annotation ; Phylogeny ; Sequence Analysis, DNA ; Species Specificity ; Virulence/genetics
Language	English
Publishing date	2011-07-19
Publishing country	United States
Document type	Comparative Study ; Journal Article ; Research Support, N.I.H., Extramural
ISSN	1932-6203
ISSN (online)	1932-6203
DOI	10.1371/journal.pone.0022420
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Article ; Online: Identification of the pangenome and its components in 14 distinct Aggregatibacter actinomycetemcomitans strains by comparative genomic analysis.

Weerayuth Kittichotirat / Roger E Bumgarner / Sirkka Asikainen / Casey Chen

PLoS ONE, Vol 6, Iss 7, p e

2011 Volume 22420

Abstract: ... for comparative genomic analysis. Two major groups were identified; serotypes a, d, e, and f, and serotypes b and ... c. A serotype e strain was found to be distinct from both groups. The size of the pangenome was 3 ...

Abstract	Aggregatibacter actinomycetemcomitans is genetically heterogeneous and comprises distinct clonal lineages that may have different virulence potentials. However, limited information of the strain-to-strain genomic variations is available.The genome sequences of 11 A. actinomycetemcomitans strains (serotypes a-f) were generated de novo, annotated and combined with three previously sequenced genomes (serotypes a-c) for comparative genomic analysis. Two major groups were identified; serotypes a, d, e, and f, and serotypes b and c. A serotype e strain was found to be distinct from both groups. The size of the pangenome was 3,301 genes, which included 2,034 core genes and 1,267 flexible genes. The number of core genes is estimated to stabilize at 2,060, while the size of the pangenome is estimated to increase by 16 genes with every additional strain sequenced in the future. Within each strain 16.7-29.4% of the genome belonged to the flexible gene pool. Between any two strains 0.4-19.5% of the genomes were different. The genomic differences were occasionally greater for strains of the same serotypes than strains of different serotypes. Furthermore, 171 genomic islands were identified. Cumulatively, 777 strain-specific genes were found on these islands and represented 61% of the flexible gene pool.Substantial genomic differences were detected among A. actinomycetemcomitans strains. Genomic islands account for more than half of the flexible genes. The phenotype and virulence of A. actinomycetemcomitans may not be defined by any single strain. Moreover, the genomic variation within each clonal lineage of A. actinomycetemcomitans (as defined by serotype grouping) may be greater than between clonal lineages. The large genomic data set in this study will be useful to further examine the molecular basis of variable virulence among A. actinomycetemcomitans strains.
Keywords	Medicine ; R ; Science ; Q
Subject code	630
Language	English
Publishing date	2011-01-01T00:00:00Z
Publisher	Public Library of Science (PLoS)
Document type	Article ; Online
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Article: Consistent intrafamilial transmission of Actinobacillus actinomycetemcomitans despite clonal diversity.

Doğan, Başak / Kipalev, Arzu Sahan / Okte, Emel / Sultan, Nedim / Asikainen, Sirkka E

Journal of periodontology

2008 Volume 79, Issue 2, Page(s) 307–315

Abstract: ... A. actinomycetemcomitans was detected in 16 of 31 (52%) family members, i.e., one parent and at least one sibling in six ...

Abstract	Background: Actinobacillus actinomycetemcomitans is a major pathogen in aggressive periodontitis. Our objectives were to determine the periodontal status and occurrence of A. actinomycetemcomitans in family members of subjects with A. actinomycetemcomitans-positive aggressive periodontitis (AgP) and to evaluate the probability of its intrafamilial transmission. Methods: Of the 300 subjects screened, 66 (22%) had AgP and A. actinomycetemcomitans. Eleven (probands) of these 66 subjects with AgP met the strict inclusion criteria for the study. The study population consisted of 55 subjects, including probands and their family members (N = 44). Two family groups were formed according to whether the proband was a child (N = 7) or a parent (N = 4). Subgingival samples from all subjects were cultured for A. actinomycetemcomitans, and its clonal types were determined by combining serotype and genotype data for each isolate. Results: Among 42 dentate family members, 16 (38%) exhibited periodontitis and eight (50%) had AgP. Periodontitis was found in nine of 12 (75%) of the dentate parents and six of 17 (35%) siblings of the child probands. A. actinomycetemcomitans was detected in 16 of 31 (52%) family members, i.e., one parent and at least one sibling in six families. The child probands shared A. actinomycetemcomitans clonal types with their parents in five of six (83%) families and with their siblings in three of six (50%) families. In the four parent-proband families, A. actinomycetemcomitans occurred in two spouses and all nine children. The parent probands shared A. actinomycetemcomitans clonal types with their spouses in both families and with their children in three of four families. In all families, the likelihood of intrafamilial transmission of A. actinomycetemcomitans was statistically significant. Members of most families (eight of 11, 73%) also harbored additional clonal types of A. actinomycetemcomitans. Conclusion: Parents and siblings of an individual with A. actinomycetemcomitans-positive AgP may have an increased susceptibility to periodontitis and shared and/or other clonal types of oral A. actinomycetemcomitans.
MeSH term(s)	Actinobacillus Infections/transmission ; Acute Disease ; Adolescent ; Adult ; Aggregatibacter actinomycetemcomitans/genetics ; Chi-Square Distribution ; Child ; Child, Preschool ; Disease Transmission, Infectious ; Family Health ; Female ; Genetic Variation ; Genotype ; Humans ; Male ; Periodontitis/microbiology ; Probability ; Serotyping
Language	English
Publishing date	2008-02
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	390921-9
ISSN	1943-3670 ; 0022-3492 ; 1049-8885 ; 0095-960X
ISSN (online)	1943-3670
ISSN	0022-3492 ; 1049-8885 ; 0095-960X
DOI	10.1902/jop.2008.070270
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Article ; Online: Vesicle-independent extracellular release of a proinflammatory outer membrane lipoprotein in free-soluble form.

Karched, Maribasappa / Ihalin, Riikka / Eneslätt, Kjell / Zhong, Deyu / Oscarsson, Jan / Wai, Sun N / Chen, Casey / Asikainen, Sirkka E

BMC microbiology

2008 Volume 8, Page(s) 18

Abstract: Background: Aggregatibacter actinomycetemcomitans is an oral bacterium associated with aggressively progressing periodontitis. Extracellular release of bacterial outer membrane proteins has been suggested to mainly occur via outer membrane vesicles. ... ...

Abstract	Background: Aggregatibacter actinomycetemcomitans is an oral bacterium associated with aggressively progressing periodontitis. Extracellular release of bacterial outer membrane proteins has been suggested to mainly occur via outer membrane vesicles. This study investigated the presence and conservation of peptidoglycan-associated lipoprotein (AaPAL) among A. actinomycetemcomitans strains, the immunostimulatory effect of AaPAL, and whether live cells release this structural outer membrane lipoprotein in free-soluble form independent of vesicles. Results: The pal locus and its gene product were confirmed in clinical A. actinomycetemcomitans strains by PCR-restriction fragment length polymorphism and immunoblotting. Culturing under different growth conditions revealed no apparent requirement for the AaPAL expression. Inactivation of pal in a wild-type strain (D7S) and in its spontaneous laboratory variant (D7SS) resulted in pleiotropic cellular effects. In a cell culture insert model (filter pore size 0.02 mum), AaPAL was detected from filtrates when strains D7S and D7SS were incubated in serum or broth in the inserts. Electron microscopy showed that A. actinomycetemcomitans vesicles (0.05-0.2 mum) were larger than the filter pores and that there were no vesicles in the filtrates. The filtrates were immunoblot negative for a cytoplasmic marker, cyclic AMP (cAMP) receptor protein. An ex vivo model indicated cytokine production from human whole blood stimulated by AaPAL. Conclusion: Free-soluble AaPAL can be extracellularly released in a process independent of vesicles.
MeSH term(s)	Aggregatibacter actinomycetemcomitans/pathogenicity ; Aggregatibacter actinomycetemcomitans/physiology ; Aggregatibacter actinomycetemcomitans/ultrastructure ; Bacterial Outer Membrane Proteins/metabolism ; Cell Culture Techniques ; Cell Membrane/ultrastructure ; Humans ; Lipoproteins/immunology ; Lipoproteins/metabolism
Chemical Substances	Bacterial Outer Membrane Proteins ; Lipoproteins
Language	English
Publishing date	2008-01-28
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ISSN	1471-2180
ISSN (online)	1471-2180
DOI	10.1186/1471-2180-8-18
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Article ; Online: Trimeric form of intracellular ATP synthase subunit β of Aggregatibacter actinomycetemcomitans binds human interleukin-1β.

Paino, Annamari / Tuominen, Heidi / Jääskeläinen, Mari / Alanko, Jonna / Nuutila, Jari / Asikainen, Sirkka E / Pelliniemi, Lauri J / Pöllänen, Marja T / Chen, Casey / Ihalin, Riikka

PloS one

2011 Volume 6, Issue 4, Page(s) e18929

Abstract: Bacterial biofilms resist host defenses and antibiotics partly because of their decreased metabolism. Some bacteria use proinflammatory cytokines, such as interleukin (IL)-1β, as cues to promote biofilm formation and to alter virulence. Although one ... ...

Abstract	Bacterial biofilms resist host defenses and antibiotics partly because of their decreased metabolism. Some bacteria use proinflammatory cytokines, such as interleukin (IL)-1β, as cues to promote biofilm formation and to alter virulence. Although one potential bacterial IL-1β receptor has been identified, current knowledge of the bacterial IL-1β sensing mechanism is limited. In chronic biofilm infection, periodontitis, Aggregatibacter actinomycetemcomitans requires tight adherence (tad)-locus to form biofilms, and tissue destroying active lesions contain more IL-1β than inactive ones. The effect of IL-1β on the metabolic activity of A. actinomycetemcomitans biofilm was tested using alamarBlue™. The binding of IL-1β to A. actinomycetemcomitans cells was investigated using transmission electron microscopy and flow cytometry. To identify the proteins which interacted with IL-1β, different protein fractions from A. actinomycetemcomitans were run in native-PAGE and blotted using biotinylated IL-1β and avidin-HRP, and identified using mass spectroscopy. We show that although IL-1β slightly increases the biofilm formation of A. actinomycetemcomitans, it reduces the metabolic activity of the biofilm. A similar reduction was observed with all tad-locus mutants except the secretin mutant, although all tested mutant strains as well as wild type strains bound IL-1β. Our results suggest that IL-1β might be transported into the A. actinomycetemcomitans cells, and the trimeric form of intracellular ATP synthase subunit β interacted with IL-1β, possibly explaining the decreased metabolic activity. Because ATP synthase is highly conserved, it might universally enhance biofilm resistance to host defense by binding IL-1β during inflammation.
MeSH term(s)	Bacterial Proteins/metabolism ; Biofilms ; Biopolymers/metabolism ; Flow Cytometry ; Humans ; Interleukin-1beta/metabolism ; Microscopy, Electron, Transmission ; Pasteurellaceae/enzymology ; Protein Binding
Chemical Substances	Bacterial Proteins ; Biopolymers ; Interleukin-1beta
Language	English
Publishing date	2011-04-18
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ISSN	1932-6203
ISSN (online)	1932-6203
DOI	10.1371/journal.pone.0018929
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Article ; Online: Vesicle-independent extracellular release of a proinflammatory outer membrane lipoprotein in free-soluble form

Oscarsson Jan / Zhong Deyu / Eneslätt Kjell / Ihalin Riikka / Karched Maribasappa / Wai Sun N / Chen Casey / Asikainen Sirkka E

BMC Microbiology, Vol 8, Iss 1, p

2008 Volume 18

Abstract: Abstract Background Aggregatibacter actinomycetemcomitans is an oral bacterium associated with aggressively progressing periodontitis. Extracellular release of bacterial outer membrane proteins has been suggested to mainly occur via outer membrane ... ...

Abstract	Abstract Background Aggregatibacter actinomycetemcomitans is an oral bacterium associated with aggressively progressing periodontitis. Extracellular release of bacterial outer membrane proteins has been suggested to mainly occur via outer membrane vesicles. This study investigated the presence and conservation of peptidoglycan-associated lipoprotein (AaPAL) among A. actinomycetemcomitans strains, the immunostimulatory effect of AaPAL, and whether live cells release this structural outer membrane lipoprotein in free-soluble form independent of vesicles. Results The pal locus and its gene product were confirmed in clinical A. actinomycetemcomitans strains by PCR-restriction fragment length polymorphism and immunoblotting. Culturing under different growth conditions revealed no apparent requirement for the AaPAL expression. Inactivation of pal in a wild-type strain (D7S) and in its spontaneous laboratory variant (D7SS) resulted in pleiotropic cellular effects. In a cell culture insert model (filter pore size 0.02 μm), AaPAL was detected from filtrates when strains D7S and D7SS were incubated in serum or broth in the inserts. Electron microscopy showed that A. actinomycetemcomitans vesicles (0.05–0.2 μm) were larger than the filter pores and that there were no vesicles in the filtrates. The filtrates were immunoblot negative for a cytoplasmic marker, cyclic AMP (cAMP) receptor protein. An ex vivo model indicated cytokine production from human whole blood stimulated by AaPAL. Conclusion Free-soluble AaPAL can be extracellularly released in a process independent of vesicles.
Keywords	Microbiology ; QR1-502 ; Science ; Q ; DOAJ:Microbiology ; DOAJ:Biology ; DOAJ:Biology and Life Sciences
Subject code	500
Language	English
Publishing date	2008-01-01T00:00:00Z
Publisher	BioMed Central
Document type	Article ; Online
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Article ; Online: Trimeric form of intracellular ATP synthase subunit β of Aggregatibacter actinomycetemcomitans binds human interleukin-1β.

Annamari Paino / Heidi Tuominen / Mari Jääskeläinen / Jonna Alanko / Jari Nuutila / Sirkka E Asikainen / Lauri J Pelliniemi / Marja T Pöllänen / Casey Chen / Riikka Ihalin

PLoS ONE, Vol 6, Iss 4, p e

2011 Volume 18929

Abstract	Bacterial biofilms resist host defenses and antibiotics partly because of their decreased metabolism. Some bacteria use proinflammatory cytokines, such as interleukin (IL)-1β, as cues to promote biofilm formation and to alter virulence. Although one potential bacterial IL-1β receptor has been identified, current knowledge of the bacterial IL-1β sensing mechanism is limited. In chronic biofilm infection, periodontitis, Aggregatibacter actinomycetemcomitans requires tight adherence (tad)-locus to form biofilms, and tissue destroying active lesions contain more IL-1β than inactive ones. The effect of IL-1β on the metabolic activity of A. actinomycetemcomitans biofilm was tested using alamarBlue™. The binding of IL-1β to A. actinomycetemcomitans cells was investigated using transmission electron microscopy and flow cytometry. To identify the proteins which interacted with IL-1β, different protein fractions from A. actinomycetemcomitans were run in native-PAGE and blotted using biotinylated IL-1β and avidin-HRP, and identified using mass spectroscopy. We show that although IL-1β slightly increases the biofilm formation of A. actinomycetemcomitans, it reduces the metabolic activity of the biofilm. A similar reduction was observed with all tad-locus mutants except the secretin mutant, although all tested mutant strains as well as wild type strains bound IL-1β. Our results suggest that IL-1β might be transported into the A. actinomycetemcomitans cells, and the trimeric form of intracellular ATP synthase subunit β interacted with IL-1β, possibly explaining the decreased metabolic activity. Because ATP synthase is highly conserved, it might universally enhance biofilm resistance to host defense by binding IL-1β during inflammation.
Keywords	Medicine ; R ; Science ; Q
Subject code	572
Language	English
Publisher	Public Library of Science (PLoS)
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article: 16S rDNA PCR-denaturing gradient gel electrophoresis in determining proportions of coexisting Actinobacillus actinomycetemcomitans strains.

Ihalin, Riikka / Asikainen, Sirkka

Journal of microbiological methods

2006 Volume 65, Issue 3, Page(s) 417–424

Abstract: ... from those of the other serotypes. Contrary to the strains of serotypes c, d, and e, strains of serotypes a, b, and f consistently ...

Abstract	Certain serotypes of Actinobacillus actinomycetemcomitans seem to prefer coexistence in vivo. The 16S rDNA PCR-denaturing gradient gel electrophoresis (DGGE) was tested for its capability to distinguish coexisting A. actinomycetemcomitans strains of different serotypes or genetic lineages and to determine their proportions in vitro. The migration pattern of the PCR amplicon from serotype c differed from those of the other serotypes. Contrary to the strains of serotypes c, d, and e, strains of serotypes a, b, and f consistently demonstrated intra-serotype migration patterns similar to each other. Since the migration patterns differed between serotype c and b strains a strain of each was used to determine their proportional representation in a strain mixture. The strains were distinguishable from each other above the 5% PCR-DGGE detection level (12.5 ng DNA/1.5 x 10(6) cells). DGGE provides a promising tool for in vitro studies on the coexistence of different genetic lineages of A. actinomycetemcomitans.
MeSH term(s)	Aggregatibacter actinomycetemcomitans/classification ; Aggregatibacter actinomycetemcomitans/genetics ; Aggregatibacter actinomycetemcomitans/growth & development ; Aggregatibacter actinomycetemcomitans/isolation & purification ; Bacterial Typing Techniques ; DNA, Bacterial/analysis ; DNA, Ribosomal/analysis ; Ecosystem ; Electrophoresis, Agar Gel/methods ; Humans ; Polymerase Chain Reaction/methods ; RNA, Ribosomal, 16S/genetics ; Serotyping ; Species Specificity
Chemical Substances	DNA, Bacterial ; DNA, Ribosomal ; RNA, Ribosomal, 16S
Language	English
Publishing date	2006-06
Publishing country	Netherlands
Document type	Evaluation Studies ; Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	604916-3
ISSN	1872-8359 ; 0167-7012
ISSN (online)	1872-8359
ISSN	0167-7012
DOI	10.1016/j.mimet.2005.08.016
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