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At least 289 records · Page 16Linked to original sources
PubMed2000

Chlamydia pneumoniae and atherosclerosis. Identification of bacterial DNA in the arterial wall.

OBJECTIVE: The intracellular Gram-negative bacterium Chlamydia pneumoniae has been associated with atherosclerosis. The presence of Chlamydia pneumoniae has been investigated in fragments of the arterial wall with a technique for DNA identification. METHODS: Arterial fragments obtained from vascular surgical procedures in 58 patients were analyzed. From these patients, 39 were males and the mean age was 65+/-6 years. The polymerase chain reaction was used to identify the bacterial DNA with a pair of primers that codify the major outer membrane protein (MOMP) of Chlamydia pneumoniae. The amplified product was visualized by electrophoresis in the 2% agarose gel stained with ethidium bromide, and it was considered positive when migrating in the band of molecular weight of the positive controls. RESULTS: Seven (12%) out of the 58 patients showed positive results for Chlamydia pneumoniae. CONCLUSION: DNA from Chlamydia pneumoniae was identified in the arterial wall of a substantial number of patients with atherosclerosis. This association, which has already been described in other countries, corroborates the evidence favoring a role played by Chlamydia pneumoniae in atherogenesis.

Aged↗
PubMed1982

Passage of bacterial DNA into host cells during in vitro transformation of Nicotiana tabacum by Agrobacterium tumefaciens.

Tumorigenic and non-tumorigenic strains of Agrobacterium tumefaciens were labelled with tritiated thymidine and added separately to suspension cultures of X-D line habituated tobacco cells. The subsequent association of bacteria with plant host cells and the passage of bacteria and bacterial DNA into these cells was examined by conventional light and electron microscopy and light microscope autoradiography. The results suggest that both strains of bacterium enter damaged host cells (via wall lesions), where they remain restricted to cell vacuoles. By 48 h, the radioactive label has spread from the vacuoles of infiltrated cells to the non-vacuolar cytoplasm of the whole callus tissue. Tumorigenic bacteria differ from the non-tumorigenic strain in promoting a significant increase in host cell nuclear volume, which is paralleled by clear and specific nuclear labelling. It is proposed that tumorigenic DNA, but not non-tumorigenic DNA, is specifically incorporated into, and activates, the host cell nucleus.

Cell Nucleus↗
PubMed2003

Inhibition of bacterial DNA replication by zinc mobilization during nitrosative stress.

Phagocytic cells inhibit the growth of intracellular pathogens by producing nitric oxide (NO). NO causes cell filamentation, induction of the SOS response, and DNA replication arrest in the Gram-negative bacterium Salmonella enterica. NO also induces double-stranded chromosomal breaks in replication-arrested Salmonella lacking a functional RecBCD exonuclease. This DNA damage depends on actions of additional DNA repair proteins, the RecG helicase, and RuvC endonuclease. Introduction of a recG mutation restores both resistance to NO and the ability of an attenuated recBC mutant Salmonella strain to cause lethal infection in mice, demonstrating that bacterial DNA replication is inhibited during host-pathogen interactions. Inhibition of DNA replication during nitrosative stress is invariably accompanied by zinc mobilization, implicating DNA-binding zinc metalloproteins as critical targets of NO-related antimicrobial activity.

Animals↗
PubMed1998

Bacterial DNA as immune cell activator.

Pattern recognition receptors of the innate and adaptive immune systems apparently recognize unmethylated CpG motifs of bacterial DNA. Cells of the innate immune system are activated directly by CpG motifs, and the resulting response dictates a Th1 bias to the developing adaptive immune response. Interestingly, antigen receptor occupancy of cells of the adaptive immune system augments their responsiveness to CpG motifs, suggesting that co-stimulatory mechanisms are operative.

Animals↗
PubMedDate not supplied

[A comparative study of different methods of determining the Tm of bacterial DNA (AUTHOR'S TRANSL)].

Melting point curves of DNA are used to calculate average GC content. Four methods of Tm determination are described and are used in the case of all DNA samples studied. The results presented confirme that thermal denaturation curves are asymetrical, and this limits the use of "normal probability paper" and "regression lines" methods. Best reproductibility is obtained by a study of the function (see article) followed by a linear interpolation of the median. This approach yields at the same time interesting information about bacterial DNA. The graphic determination of Tm appears to be the method for choice for routine analysis.

Citrobacter↗
PubMed1976

[Differences in the mutagenic effectiveness of N-nitrosomethyl- and N-nitrosoethylurea acting on bacterial DNA in vivo].

Chemically induced lethals and reversion frequencies of two mutants of Salmonella typhimurium LT-2 1530 and 1532 were examined. Prototrophic mutants to histidine independence occurred by two different mechanisms and were induced by nitrous acid, hydroxylamine, N-nitrosomethylurea (NMU), N-nitrosoethylurea (NEU) and N, N, nitrosodimethylurea (NDMU). The ability of the mutants to revert was affected by the chemical structure (in the row of N-nitrosoalkylurea - by alkyl group) of the mutagen. NEU is found to induce in bacterial DNA only base pair substitutions; NMU and NDMU - deletions and base pair substitutions. An original model of deletion mechanism with Goldwait endonuclease II participation is proposed.

Base Sequence↗
PubMed1999

The actions of bacterial DNA on murine macrophages.

Murine macrophages are able to distinguish bacterial from mammalian DNA. The response is mimicked by single-stranded oligonucleotides containing unmethylated CG dinucleotides ("CpG" motifs) in specific sequence contexts. The dose-response curve for activation is influenced by variation in the sequence flanking the core CpG motif. CpG or bacterial DNA activates several signaling pathways in common with bacterial lipopolysaccharide (LPS), leading to induction of cytokine genes such as tumor necrosis factor alpha. Pretreatment with LPS causes desensitization to subsequent activation by CpG DNA. Both stimuli also cause cell cycle arrest in macrophages proliferating in response to the macrophage growth factor colony-stimulating factor-1 (CSF-1), but prevent apoptosis caused by growth factor removal. In part, cell cycle arrest by CpG DNA and LPS may be linked to rapid down-modulation of the CSF-1 receptor from the cell surface, a response that occurs in an all-or-nothing manner. The response of macrophages to CpG DNA has aspects in common with the DNA damage response in other cell types, which may provide clues to the underlying mechanism.

Animals↗
PubMed2000

Bacterial DNA is implicated in the inflammatory response to delivery of DNA/DOTAP to mouse lungs.

Phase 1 clinical trials of liposome-mediated gene therapy for cystic fibrosis have been completed and in all cases the expression level achieved has been low and transient. Clearly, improvements in the efficiency of gene transfer are required. It is now being recognised that delivery of high doses of DNA/liposomes to the mouse airway epithelium can achieve reproducible evidence of transgene, but is often associated with an unacceptable level of inflammation/ toxicity. It has recently been shown that instillation of bacterial DNA causes inflammation in the lower respiratory tract of rodents. The increased number and unmethylated status of CpG motifs, particularly when present in a particular base context, was identified as an important factor in this response. It was suggested that the immune system recognises this molecular pattern as 'foreign' thus activating appropriate immune responses. We have found that methylation of DNA decreases the level of several inflammatory cytokines in lavage fluid and surprisingly has a differential effect on expression of the plasmids pCMV CFTR-int6ab and pCMV CAT which only differ in the actual transcription cassette. The severe lung pathology observed did not show a corresponding decrease with methylation suggesting that these cytokines are not the only contributors to the toxicity/inflammation observed. Gene Therapy (2000) 7, 384-392.

Animals↗
PubMed1999

FramePlot: a new implementation of the frame analysis for predicting protein-coding regions in bacterial DNA with a high G + C content.

FramePlot is a web-based tool for predicting protein-coding regions in bacterial DNA with a high G + C content, such as Streptomyces. The graphical output provides for easy distinction of protein-coding regions from non-coding regions. The plot is a clickable map. Clicking on an ORF provides not only the nucleotide sequence but also its deduced amino acid sequence. These sequences can then be compared to the NCBI sequence database over the Internet. The program is freely available for academic purposes at http://www.nih.go.jp/jun/cgi-bin/frameplot.pl.

Amino Acid Sequence↗
PubMed1995

[Aseptic meningitis. Demonstration of bacterial DNA in cerebrospinal fluid by gene amplification].

OBJECTIVE: To develop a diagnostic tool to recognize whether a postoperative meningitis occurring in neurosurgical patients is of bacteriological origin or not, in detecting in CSF bacterial DNA with the polymerase chain reaction (PCR) technique. STUDY DESIGN: Laboratory study. PATIENTS: Twenty-seven neurosurgical ICU patients associating, in the postoperative period, the CDC criteria of meningitis and a neutrophil polymorphonuclear count over 100 cells.mm-3 were allocated either into the MB+ group (n = 7) when their CSF culture was positive or in the MB- group (n = 20) when the culture was sterile. The CSF of 43 neurosurgical ICU patients without postoperative clinical and biological features of meningitis acted as controls. Sixteen specimens out of the 43 were inoculated with bacteria at a known concentration. METHODS: The CSF specimens of all patients were tested for the presence of eurcaryote DNA using the PCR technique. Beforehand its sensitivity had been assessed using the inoculated CSF of control group: a positive amplification at 20 cycles was equivalent to 10(5) CFU.mL-1 and a positive amplification at 25 cycles to 10(3) CFU.mL-1. RESULTS: In the 43 sterile control CSF specimens the amplification was negative in all at 20 cycles and in 42 at 25 cycles. In the 16 previously sterile control specimens supplemented with bacteria, as well as in the CSF of all 7 patients of MB+ group the amplification was positive at 20 and 25 cycles. In those of MB- group the amplification was negative in all at 20 cycles, but was positive in 19 out of 20 at 25 cycles. Southern blot with specific procaryote probes was positive with amplification products from CSF of MB+ and MB- groups and negative with control CSFs and human DNA. DISCUSSION: The presence of bacteria in CSF of patients sustaining a meningitis can be accurately detected through their DNA. Postoperative aseptic meningitides may have a bacterial origin. PCR can be used as a routine technique to provide a diagnosis of bacterial meningitis in less than 6 hours. Additionally specific oligonucleotides allow to identify the bacteria in less than 12 hours.

Adolescent↗
PubMed1992

Release of bacterial DNA by marine nanoflagellates, an intermediate step in phosphorus regeneration.

The concentrations of dissolved DNA and nanoflagellates were found to covary during a study of diel dynamics of the microbial food web in the Adriatic Sea. This observation was further investigated in a continuous seawater culture when nanoflagellates were fed bacteria grown in filtered seawater. Analysis of dissolved organic phosphorus and dissolved DNA showed a sixfold increase of dissolved DNA in the presence of the nanoflagellates (Ochromonas sp.). The amount of DNA released suggested that the majority of the consumed bacterial DNA was ejected. Phagotrophic nanoflagellates thus represent an important source of origin for dissolved DNA. The rate of breakdown of dissolved DNA and release of inorganic phosphorus in the pelagic ecosystem is suggested to be dependent on the ambient phosphate pool. In the P-limited northern Adriatic Sea, rapid degradation of the labelled DNA could be demonstrated, whereas the N-limited southern California bight water showed a much lower rate. Phosphorus originating from dissolved DNA was shown to be transferred mainly to organisms in the <3-mum-size fractions. On the basis of the C/P ratios, we suggest that a significant fraction of the phosphorus demand by the autotrophs may be sustained by the released DNA during stratified conditions. Thus, the nucleic acid-rich bacterial biomass grazed by protozoa plays an important role in the biogeochemical cycling of phosphorus in the marine environment.

Journal Article↗
PubMed2006

Detection of bacterial DNA in atheromatous plaques by quantitative PCR.

This is the first study to analyze atheromatous plaques for the presence of bacterial DNA from ten species, including periodontal species and Chlamydia pneumoniae. We examined 129 samples of DNA extracted from atheromas from 29 individuals for the presence of bacterial 16S rDNA sequences from ten different species: Porphyromonas gingivalis, Actinobacillus actinomycetemcomitans (A.a.), Tannerella forsythensis, Eikenella corrodens, Prevotella intermedia, Staphylococcus aureus, Staphylococcus epidermidis, Streptococcus mutans, Treponema denticola and C. pneumoniae. All determinations were made using real-time quantitative polymerase chain reaction (PCR) methods employing SYBR Green. Species from the Bacteroides family were found in about 17% of the young but approximately 80% in elderly patients. Almost half of the samples contained DNA from A. a. and C. pneumoniae, although the proportion of the latter was minimal. S. aureus and S. epidermidis were found with the lowest frequency, 5 and 10%, respectively. S. mutans was found in approximately 20% of the samples. The proportions of each bacterial species were calculated relative to the total amount of prokaryotic DNA. The data support our previous findings of an association between periodontal organisms and vascular inflammation. We conclude that DNA from oral infectious agents is commonly found in atheromas from young but especially from elderly subjects, and that the contribution of C. pneumoniae to the inflammation may be minimal.

Adolescent↗
PubMed2000

The major role of macrophages and their product tumor necrosis factor alpha in the induction of arthritis triggered by bacterial DNA containing CpG motifs.

OBJECTIVE: To understand the mechanisms of arthritis triggered by CpG-containing oligonucleotides (ODN). METHODS: Following the induction of CpG ODN-triggered arthritis in mice, we analyzed the impact of depletion of immune cells, including neutrophils, natural killer (NK) cells, and monocyte/macrophages, on the arthritis, as well as the impact in SCID mice lacking T and B cells. In addition, tumor necrosis factor alpha (TNFalpha) knockout mice were studied, and intraarticular administration of p65 antisense to nuclear factor kappaB (NF-kappaB) was used to examine effects in CpG ODN-triggered arthritis. Cytokine messenger RNA expression in synovial tissue was evaluated by in situ hybridization. RESULTS: The presence of macrophages was mandatory for the mediation of arthritis triggered by CpG ODN, whereas the absence of neutrophils, NK cells, T cells, and B cells was of minor importance in this regard. The proinflammatory cytokines TNFalpha, interleukin-1beta, and interleukin-12, which originate from macrophages, were frequently found in the inflamed joints, and TNFalpha was confirmed to be an important mediator in the development of arthritis, since the incidence and severity of joint inflammation were markedly reduced in TNFalpha knockout mice. NF-kappaB exerted an important regulatory role in the development of CpG ODN-mediated arthritis, since local administration of antisense to the p65 subunit of NF-kappaB diminished the incidence of inflammation by 50%. CONCLUSION: Macrophages and their products play an important role in the development of arthritis triggered by bacterial DNA containing CpG motifs.

Animals↗
PubMed1988

The relationship between the base composition of bacterial DNA and its intracellular melting temperature as determined by differential scanning calorimetry.

The correlation between the melting temperature of intracellular DNA, determined by differential scanning calorimetry (DSC) of whole bacteria, and its guanine + cytosine (G + C) content, was examined for 58 species of bacteria. Samples of vegetative cells were heated in a Perkin-Elmer DSC-2C at 10 degrees C min-1 from 5 to 130 degrees C, cooled to 5 degrees C and then re-heated as before. Literature values for the mole fraction of G + C, XGC, were linearly related to the temperature, Tmax, at which the reversible peak, pr, observed on the second heating run was at a maximum, via the equation XGC = (Tmax -73.8)/41.0. This equation accounted for 91.9% of the variance in XGC with 95% confidence limits of +/- 7.3%, approximately 1.6 times the corresponding uncertainty (+/- 4.5%) quoted by De Ley (Journal of Bacteriology 101, 738-754, 1970) for estimates based on the spectroscopically determined melting temperature of purified DNA. Random errors of measurement of Tmax did not greatly limit the precision of the prediction and it was concluded that factors additional to base composition affected the temperature of DNA melting within the bacterial cell. Displacement of Tmax values from the fitted line was particularly noticeable in Campylobacter, Corynebacterium and Bacterionema species and part of the residual variation appeared to be species specific, possibly caused by differences in intracellular solute concentration.

Base Composition↗
PubMed2004

Silencing of episomal transgene expression by plasmid bacterial DNA elements in vivo.

We previously demonstrated that sustainable enhanced levels of transgene products could be expressed from a bacterial DNA-free expression cassette either formed from a fragmented plasmid in mouse liver or delivered as a minicircle vector. This suggested that bacterial DNA sequences played a role in episomal transgene silencing. To further understand the silencing mechanism, we systematically altered the DNA components in both the expression cassette and the bacterial backbone, and compared the gene expression profiles from mice receiving different DNA forms. In nine vectors tested, animals that received the purified expression cassette alone always expressed persistently higher levels of transgene compared to 2fDNA groups. In contrast, animals that received linearized DNA by a single cut in the bacterial backbone had similar expression profiles to that of intact plasmid groups. All three linear DNAs formed large concatemers and small circles in mouse liver, while ccDNA remained intact. In all groups, the relative amount of vector DNA in liver remained similar. Together, these results further established that the DNA silencing effect was mediated by a covalent linkage of the expression cassette and the bacteria DNA elements.

Animals↗
PubMed1996

Mode of action of GR122222X, a novel inhibitor of bacterial DNA gyrase.

GR122222X is a potent inhibitor of the supercoiling reaction of bacterial DNA gyrase. We show that this compound binds stoichiometrically to inactivate the ATPase activity of a 43-kDa N-terminal fragment of the B subunit and competitively inhibits the binding of a radiolabelled coumarin drug to N-terminal fragments of GyrB. These and other data suggest that GR122222X has a mode of action similar, but not identical, to that of coumarin antibiotics.

DNA Topoisomerases, Type II↗
PubMed2005

AFM studies on the role of the protein RdgC in bacterial DNA recombination.

Genetic studies of rdgC in different bacterial systems suggest that it may play a role in replication and recombination. However, the exact function of the corresponding protein, RdgC, is unknown. In this study, we have imaged complexes of RdgC with both linear and supercoiled circular plasmid DNA using atomic force microscopy. We confirm that RdgC does not target any specific sequences in double-stranded DNA, as has been suggested from biochemical data. However, we detect an increased affinity of the protein to DNA ends, and an ability to promote bending of DNA. Similar binding preferences have been reported for enzymes involved in recombination. Protein complexes with supercoiled plasmid DNA further enabled us to study the effect of RdgC on DNA superstructure. At high concentrations of protein we observed promotion of DNA condensation. Recombination is largely enhanced by close contacts of distant regions along the DNA strands, as can occur, for instance, through condensation. Our data thus support a possible function of RdgC as a midwife of recombination.

DNA, Bacterial↗