Fimbriae Proteins ; genetics ; Genotype ; Humans ; Porphyromonas gingivalis ; genetics ; pathogenicity ; Virulence

OBJECTIVETo investigate the pathogenicity of matrix metalloproteinase 8, 9 (MMP-8, MMP-9) regulations of polymorphonuclear leukocytes (PMNs) by challenge of Porphyromonas gingivalis (P. gingivalis) with different fimA genotypes.

METHODSThe studies mainly adopt the isopycnic sedimentation separation to separate the PMNs from human peripheral blood. P. gingivalis ATCC 33277 (type I), WCSP 115 (type II), WCSP 1.5 (type III), W83 (type IV), WCSP 559 (type IV) were assessed for their inductions of MMP-8, MMP-9 expression in PMNs. MMP-8, MMP-9 protein levels in culture supernatant were determined by ELISA at different time intervals (5 min, 30 min, 1 h, 2 h) following continuous co-culture of bacteria with PMNs.

RESULTSMMP-8 and MMP-9 protein levels produced by PMNs co-culture with the I fimA-IV fimA P. gingivalis were significantly stronger than unsimulated group. The velocity and quantity of MMP-8 produced by PMNs co-culture with the II fimA P. gingivalis and IV fimA P. gingivalis were more than III fimA, IVfimA P. gingivalis. The MMP-9 protein levels produced by PMNs co-culture with the I fimA, II fimA, IV fimA P. gingivalis was significantly stronger than III fimA and IV fimA P. gingivalis.

CONCLUSIONII fimA and IV fimA P. gingivalis have stronger pathogenicity relatively, which indicate that fimA genotype is associated with pathogenesis of P. gingivalis.

Coculture Techniques ; Fimbriae Proteins ; Genotype ; Humans ; Matrix Metalloproteinase 8 ; Neutrophils ; Porphyromonas gingivalis

UNLABELLEDOBJECTIVE; To clone the fimA gene of Porphyromonas gingivalis (P. gingivalis) and detect its expression in Escherichia coli (E. coli).

METHODSThe fimA gene was obtained by PCR from the genome of P. gingivalis to construct a prokaryotic expression plasmid pT-BAD/fimA. pT-BAD/fimA was transformed into E. coli BL21 (DE3) competent cells and the recombination protein was characterized by means of matrix assisted laser desorption ionisation time-of-flight mass spectrometry (MALDI-TOF-MS) analysis. The bound protein was eluted with different concentrations of imidazole (250, 200, 150, 100, 50 micromol x L(-1)) respectively.

RESULTSDNA sequencing showed that the fragment was 99.9% consistent with that of the published. After induction with L-arabinose, a new 3.8 x 10(4) protein appeared on SDS-PAGE gel. The protein was further identified by MALDI-TOF-MS. Purity of 95% of the target protein was purified by Ni-NTA Purification System after eluted with 100 micromol x L(-1) imidazole.

CONCLUSIONThe fimA gene of P. gingivalis was cloned successfully and its protein was expressed correctly in E. coli. A high purity of protein FimA was obtained and it could be applied for follow-up researches.

Cloning, Organism ; Escherichia coli ; Fimbriae Proteins ; Plasmids ; Polymerase Chain Reaction ; Porphyromonas gingivalis

As an important protein structure on the surface of bacteria, type Ⅳ pili (TFP) is the sensing and moving organ of bacteria. It plays a variety of roles in bacterial physiology, cell adhesion, host cell invasion, DNA uptake, protein secretion, biofilm formation, cell movement and electron transmission. With the rapid development of research methods, technical equipment and pili visualization tools, increasing number of studies have revealed various functions of pili in cellular activities, which greatly facilitated the microbial single cell research. This review focuses on the pili visualization method and its application in the functional research of TFP, providing ideas for the research and application of TFP in biology, medicine and ecology.
Fimbriae, Bacterial/metabolism* ; Bacterial Proteins/genetics* ; Bacterial Physiological Phenomena ; Bacterial Adhesion/physiology*

OBJECTIVETo investigate the distribution of fimA genotype of P. gingivalis in chronic periodontitis patients.

METHODSSubgingival plaque samples were collected from 101 chronic periodontitis patients. P. gingivalis was detected by both culture method and P. gingivalis 16S rRNA PCR. fimA type-specific primer were designed, and the distribution of fimA genotype of P. gingivalis in periodontitis patients were detected by PCR.

RESULTSThe detective ratio of P. gingivalis was 88.1%. Among them, a single fimA genotype was detected in most subgingival plaque samples (65.1%), and the distribution of five fimA genotypes among P. gingivalis positive patients was as follows: type I, 24.7%; type II, 43.8%; type III, 15.7%; type IV, 40.4%; type V, 3.4%; respectively.

CONCLUSIONP. gingivalis with various fimA genotypes were present in subgingival plaque samples from chronic periodontitis patients, and P. gingivalis with type II fimA and IV fimA were more predominant in chronic periodontitis patients, and they may be associated with the development of periodontitis.

Adult ; Chronic Periodontitis ; microbiology ; Dental Plaque ; Female ; Fimbriae Proteins ; genetics ; Genotype ; Humans ; Male ; Periodontitis ; Polymerase Chain Reaction ; Porphyromonas gingivalis ; genetics

OBJECTIVETo construct the recombinant plasmid pPHU281_A_Spec_B, which knock out Porphyrmonas gingivalis (Pg) FimA gene.

METHODSGenomic DNA was extracted from PgATCC33277 which was cultured in anaerobic condition. The upstream and downstream gene of FimA was cloned from Pg genenomic DNA with specific restriction sites by polymerase chain reaction. Suicide vector pPHU281 was inserted by three fragments: upstream, downstream of FimA gene and spectinomycin resistance gene. The recombinant plasmid was confirmed by electrophoresis and sequenced after amplification in compentent cells DH-5α.

RESULTSThe gene sequence was identified by DNA sequencing analysis. The recombinant plasmid pPHU281_A_Spec_B was successfully constructed.

CONCLUSIONSThe recombinant plasmid pPHU281_A_Spec_B was constructed, which may be used for the constructon of FimA deficient Pg.

Base Sequence ; DNA, Bacterial ; genetics ; Fimbriae Proteins ; genetics ; Gene Knockout Techniques ; Genes, Bacterial ; Genetic Vectors ; Plasmids ; genetics ; Porphyromonas gingivalis ; genetics ; Recombinant Proteins ; genetics ; Sequence Analysis, DNA

Adhesins, Escherichia coli ; genetics ; Bacterial Typing Techniques ; Escherichia coli Proteins ; genetics ; Fimbriae Proteins ; genetics ; Genes, Bacterial ; Genotype ; Uropathogenic Escherichia coli ; classification ; genetics

To date, efforts to treat autoimmune diseases have primarily focused on the disease symptoms rather than on the cause of the disease. In large part, this is attributed to not knowing the responsible auto-antigens (auto-Ags) for driving the self-reactivity coupled with the poor success of treating autoimmune diseases using oral tolerance methods. Nonetheless, if tolerogenic approaches or methods that stimulate regulatory T (Treg) cells can be devised, these could subdue autoimmune diseases. To forward such efforts, our approach with colonization factor antigen I (CFA/I) fimbriae is to establish bystander immunity to ultimately drive the development of auto-Ag-specific Treg cells. Using an attenuated Salmonella vaccine expressing CFA/I fimbriae, fimbriae-specific Treg cells were induced without compromising the vaccine's capacity to protect against travelers' diarrhea or salmonellosis. By adapting the vaccine's anti-inflammatory properties, it was found that it could also dampen experimental inflammatory diseases resembling multiple sclerosis (MS) and rheumatoid arthritis. Because of this bystander effect, disease-specific Treg cells are eventually induced to resolve disease. Interestingly, this same vaccine could elicit the required Treg cell subset for each disease. For MS-like disease, conventional CD25+ Treg cells are stimulated, but for arthritis CD39+ Treg cells are induced instead. This review article will examine the potential of treating autoimmune diseases without having previous knowledge of the auto-Ag using an innocuous antigen to stimulate Treg cells via the production of transforming growth factor-beta and interleukin-10.
Animals ; Antigens, Bacterial/*immunology ; Arthritis, Rheumatoid/immunology/prevention & control ; Autoantigens/*immunology ; Fimbriae Proteins/*immunology ; Humans ; Multiple Sclerosis/immunology/prevention & control ; Salmonella/*immunology ; T-Lymphocytes, Regulatory/*immunology ; *Vaccination

OBJECTIVETo detect the distribution of fimA genotype of P. gingivalis in periodontally healthy adults and chronic periodontitis patients, and to investigate the relationship between the prevalence of fimA genotype of P. gingivalis and periodontal health status.

METHODSSubgingival plaque samples were collected from 136 periodontally healthy adults and 115 chronic periodontitis patients. The occurrence of P. gingivalis was determined by P. gingivalis 16S rRNA PCR. Distribution of fimA genotype was assessed in P. gingivalis positive samples by PCR using primers pairs homologous to the different fimA genes.

RESULTSP. gingivalis was detected in 22.1% of the healthy subjects and 81.7% of chronic periodontitis patients. A single fimA genotype was detected in most subgingival plaque samples. In P. gingivalis-positive healthy adults, the most prevalent fimA genotype of P. gingivalis was type I fimA. In contrast, a majority of chronic periodontitis patients carried type II fimA, followed by IV fimA and I b fimA. The univariate analysis illustrated that chronic periodontitis was associated with occurrences of type I fimA (OR = 0.97), I b (OR =13.26), II (OR = 36.62), III (OR = 4.57), IV (OR = 22.86), and V (OR = 1.19).

CONCLUSIONII fimA genotype of P. gingivalis followed by IV and I b were an important virulence factor that may account for the pathogenesis of chronic periodontitis, suggesting an increased pathogenic potential of these types.

Adult ; Chronic Periodontitis ; Dental Plaque ; Female ; Fimbriae Proteins ; Genotype ; Health Status ; Humans ; Male ; Periodontitis ; Polymerase Chain Reaction ; Porphyromonas gingivalis ; Prevalence ; RNA, Ribosomal, 16S

We have compared the findings of MRI with those of hip arthroscopy in 22 hips in adults with pilinful hip diseases. From March l995 to March 1997. We performed arthroscopy of the hips in symptomatic 22 patients. Fifteen cases were men and the mean average age was 46 years old. Preoperatively, the patients was diagnoseil with careful physical examinations and MR arthrogram or MR1. Among them, acetahular lahrum tears were 13 cases. synovial lesions were 3 cases and avascular necrosis of femoral head were 6 casee. We have examed cartilage softening, indentation or defect of femoral head and acetabulum, acetabular lahrum, loose hodies and synovial lesion hy arthroscopy. Under the arthroscopic guide, We had resected the torn Jahrum in S cases, removed a loose hodies such as synovial chondromatosis in 1 case and excised a synovial pathology in 2 cases. The others had a open arthrotomy in 8 cases. The sensitivity of MRI was 89.5% and its accuracy was 77.3% compared with arthroscopic findings. Average follow-up lengths was 8.6 months and 17(77.3%) of 22 patients were improved clinically. The arthroscopy of the hip is less invasive and a useful investigation for painful hip diseases for labral tears and synovial lesions in adults cspecially when standard MRI fails to prove a exact hip joint pathology, but it may be useless in avascular necrosis of femoral head. Also it is technical demanding and necessary to long-term follow-up.
Acetabulum ; Adult ; Arthroscopy ; Cartilage ; Chondromatosis, Synovial ; Diagnosis* ; Fimbriae Proteins ; Follow-Up Studies ; Head ; Hip Joint ; Hip* ; Humans ; Magnetic Resonance Imaging ; Male ; Middle Aged ; Necrosis ; Pathology ; Physical Examination