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Prevalence of HIV-1/2, HTLV-I/II, hepatitis B virus (HBV), hepatitis C virus (HCV), Treponema pallidum and Trypanosoma cruzi among prison inmates at Manhuaçu, Minas Gerais State, Brazil.

The purpose of this study was to determine the seroprevalence of human immunodeficiency virus (HIV-(1/2)), human T-cell lymphotropic virus (HTLV-I/II), hepatitis B virus (HBV), hepatitis C virus (HCV), Treponema pallidum and Trypanosoma cruzi among 63 male prisoners in Manhuaçu, Minas Gerais, Brazil and to compare this with data from eligible blood donors. The positive results were as follows: 11/63 (17.5%) for HBV, 5/63 (7.4%) for syphilis, 4/63 (6.3%) for HCV, 3/63 (4.8%) for Chagas' disease, 2/63 (3.2%) for HIV-1/2 and 1/63 (1.6%) for HTLV-I/II. The seroprevalence in prisoners was higher than among blood donors, mainly for antibodies to HIV-1/2, HCV and HBV. This is probably due to low social economic level, illiteracy, higher proportion with a prior history of intravenous drug use and/or unsafe sexual behavior. Therefore, these prisoners constitute a high-risk group and routine screening and counseling are recommended.

Adult↗

Comparison of a recombinant-antigen enzyme immunoassay with Treponema pallidum hemagglutination test for serological confirmation of syphilis.

A recombinant-antigen enzyme immunoassay (EIA), BioSCREEN anti-Treponema pallidum, was compared favorably with the T. pallidum hemagglutination test, in the detection of specific antibodies in different groups of sera from patients with primary (n = 38), secondary (n = 10), early latent (n = 28) and congenital syphilis (n = 2), patients with leptospirosis ( n= 8), infectious mononucleosis (n = 7), hepatitis (n = 9), diabetes mellitus (n = 11), rheumatoid arthritis (n = 13), leprosy (n = 11), tuberculosis (n = 9), HIV/Aids ( n= 12), systemic lupus erythematosus (n = 4), rheumatic fever (n = 3), old-persons (n = 9), pregnant women (n = 29) and blood donors (n = 164). The coincidence between them was 95.1%. The sensitivity and specificity of the EIA were 93.3% and 95.5%, respectively. Fifteen serum specimens belonging to old-persons, pregnant women, blood donors, and patients with human leptospirosis, hepatitis, diabetes mellitus, tuberculosis and rheumatic fever gave false-positive results by Venereal Disease Research Laboratory and/or Rapid Plasma Reagin. The EIA can be used as alternative method for the serological confirmation of syphilis.

Antigens, Bacterial↗

Effects of the inorganic salts sodium chloride, sodium bicarbonate, and magnesium sulfate upon the growth and motility of Treponema vincentii.

The use of inorganic salt solutions as chemotherapeutic agents in the control of periodontal disease has received considerable attention in the past few years. Although some research has been published on their clinical effectiveness, little is known about their therapeutic activity and bactericidal effects upon oral spirochetes. The present study investigated the effects of varied concentrations of NaCl, NaHCO3, and MgSO4 upon the in vitro growth and motility of Treponema vincentii. Growth determinations were performed using a turbidiometric analysis at 545 nm. Motility was qualitatively studied by direct examination of 200 treponemes in a wet mount specimen. Samples were taken at 24, 48, 72, and 96 hours following inoculation with the treponemes. Concentrations of 0.5 M NaCl, NaHCO3, or MgSO4 totally inhibited the growth and motility of T. vincentii over a 96-hour period. Salt concentrations less than or equal to 0.10 M had little if any effect upon growth and motility. The data support the hypothesis that the bactericidal and antimotility effects of these salts are related more to their concentrations than to the presence of a specific inorganic ion. They also suggest that motility may be a valid indicator of bacterial viability. Before the clinical significance of the results can be ascertained, human studies are needed to establish sulcular salt concentrations which can be achieved with local irrigation and to determine how long bactericidal concentrations can be maintained.

Bicarbonates↗

Relative proportions of pathogen-related oral spirochetes (PROS) and Treponema denticola in supragingival and subgingival plaque from patients with periodontitis.

The purpose of this study was to use monoclonal antibodies to enumerate spirochetes in dental plaque, including the newly recognized pathogen-related oral spirochete (PROS) and specific serovars of Treponema denticola. Plaque was collected from control subjects with no apparent periodontal disease and from sites of moderate to severe chronic periodontitis in patients with inflammatory periodontal disease. Individual monoclonal antibodies were used to determine whether spirochetes were present and then a double-staining protocol was employed to count total spirochetes and specific treponemes in individual microscopic fields. Results indicate that spirochetes are more common at diseased sites and in subgingival plaque than at healthy sites or in supragingival plaque. Together PROS and T. denticola comprised the majority of all spirochetes in all samples and PROS and T. denticola serovars "B" and D were most numerous in plaque from patients with periodontitis. PROS were the majority of all spirochetes in supragingival plaque (76.2% +/- 23.8%) and subgingival plaque (60.9% +/- 19.1%) from periodontitis patients, significantly larger than the percentage of T. denticola serovar "B" (P less than .001 for both supragingival and subgingival plaque) and serovar D (P less than .01 for supragingival and P less than .001 for subgingival plaque). These observations indicate that PROS are the predominant spirochete in plaque from sites of patients with periodontitis, but other analytical approaches are necessary to determine if PROS or T. denticola are pathogenic.

Adult↗

Treponema denticola and Porphyromonas gingivalis as prognostic markers following periodontal treatment.

Subgingival plaque samples were collected from individuals with advanced periodontitis before and 3 to 11 weeks after scaling and root planing periodontal treatment. The plaque levels of Treponema denticola and Porphyromonas gingivalis antigens were measured before and after treatment by a quantitative immunoassay procedure using monoclonal antibodies specific for these oral bacteria. A decrease in mean levels of T. denticola (P less than .05) and P. gingivalis antigens (P less than .09) were observed following periodontal therapy. Improved health, as measured by a decrease in probing depth, was associated with a decrease in T. denticola antigen (P less than .05). These results suggest that the T. denticola levels of successfully treated sites decreased, while non-responding sites had levels of this microbial marker which were equal to or greater than the pre-treatment levels. These results provide additional evidence that T. denticola is associated with human adult severe periodontal disease, and can serve as a prognostic marker for disease recurrence.

Adult↗

Occurrence of Porphyromonas gingivalis, Bacteroides forsythus, and Treponema denticola in periodontally healthy and diseased subjects as determined by an ELISA technique.

The aim of this study was to assess by means of an ELISA technique, the occurrence of 3 putative periodontopathogens, Porphyromonas gingivalis, Bacteroides forsythus, and Treponema denticola, in 3 clinically-defined adult periodontal conditions. Thirty systemically-healthy subjects were selected and grouped into 3 categories according to their periodontal health: 1) 10 periodontitis subjects (PS), having moderate adult chronic periodontitis; 2) 10 untreated gingivitis subjects (UGS), exhibiting no signs of periodontal destruction but presenting with clinical signs of mild gingivitis; and, 3) 10 treated gingivitis subjects (TGS), having the same clinical status as UGS, but who received a thorough prophylaxis treatment within the past 7 to 14 days prior to the baseline examination. A total of 60 samples were collected subgingivally from the six Ramfjord teeth per subject in each group and ELISA analysis was carried out to give a semiquantitative estimate of P. gingivalis. B. forsythus, and T. denticola. The immunologic detection method suggested the presence of antigens of P. gingivalis, B. forsythus, and T. denticola in subjects from each of the 3 groups. When a global analysis for the 3 disease groups was performed at one time, statistically significant differences were found among the ELISA scores of the 3 bacterial species. For example, comparisons of the ELISA scores showed that the concentrations of P. gingivalis differed significantly when comparing TGS to UGS and PS, but not when examining UGS/PS. The ELISA scores for B. forsythus were significantly different between TGS and PS. Mean concentrations of T. denticola were significantly different when comparing PS to TGS or UGS, whereas no difference was found between the latter categories. Within the limited scope of this study, the concentration of antigens detectable from putative periodontopathogens like P. gingivalis, B. forsythus, and T. denticola differed among the 3 diseased groups, with periodontitis subjects often showing the greatest level of antigens. Thus, it is reasonable to expect that, when using sensitive immunological detection methods, antigens of suspected periodontal pathogens can be found irrespective of the individual's clinical status. However, while detectable in the periodontal sites, the concentrations of these microorganisms are most likely to be above the threshold necessary to induce clinically-significant disease. Studies with larger sample size and standardized antigens are necessary to determine if the groups we found not to differ, were, in fact, different.

Adult↗

Effects of whole cell sonicates of Treponema lecithinolyticum on osteoclast differentiation.

BACKGROUND: Alveolar bone destruction is a characteristic feature of periodontal diseases and multinucleated osteoclast cells derived from hemopoietic cells are responsible for bone resorption. Treponema lecithinolyticum is a novel oral spirochete isolated from the periodontal lesions. METHODS: The effect of whole cell sonicates on the osteoclast differentiation was examined in a co-culture system of hemopoietic mouse bone marrow cells and calvaria derived-osteoblastic cells to clarify the role of T. lecithinolyticum in the alveolar bone destruction associated with periodontal diseases. The differentiated osteoclasts were confirmed by tartrate-resistant acid phosphatase (TRAP) staining. RESULTS: Sonicates of this bacterium stimulated the osteoclast formation in the co-culture system in a dose-dependent manner. The sonicates-induced osteoclast formation was partially inhibited by the heat treatment of sonicates. Indomethacin, which is a prostaglandin inhibitor, decreased the osteoclast formation induced by the bacterial sonicates. CONCLUSIONS: These findings suggest that T. lecithinolyticum induces osteoclast differentiation by a prostaglandin E2-dependent mechanism and that heat-labile components may be involved in this process.

Alveolar Bone Loss↗

Activation of matrix metalloproteinase-2 by a novel oral spirochetal species Treponema lecithinolyticum.

BACKGROUND: Periodontal tissue destruction is a characteristic of periodontitis. This can be caused by either bacterial enzymes or host cell-derived matrix metalloproteinases (MMPs). In order to elucidate the etiologic role of oral spirochetes, we investigated the effects of Treponema lecithinolyticum, a novel saccharolytic species, on MMP-2 activation. METHODS: Gingival fibroblasts (GFs) and periodontal ligament (PDL) cells obtained from healthy human subjects were cultured to confluence in alpha-minimal essential medium (alpha-MEM) supplemented with 10% fetal bovine serum. After serum starvation for a day, the cultures were treated with whole cell sonicates, heat-denatured whole cell sonicates, outer membrane fraction (OMF) or formaldehyde-fixed cells of T. lecithinolyticum. Culture supernatants were collected after incubation for 24 to 48 hours and analyzed for MMP-2 activation by gelatin zymography. Collagenolytic activity was quantitatively measured using human [3H] type IV collagen as a substrate. RESULTS: Treatment of GFs and PDL cells with whole cell sonicates, formaldehyde-fixed whole cells, or the OMF of T. lecithinolyticum resulted in the production of MMP-2 partly in the fully active form with a molecular mass of 62 kDa, whereas non-treated control cultures and cultures treated with a heat-denatured fraction did not show the active form. Cultures exposed to T. lecithinolyticum had higher collagenolytic activity than non-treated cultures. CONCLUSIONS: Our results demonstrate that T. lecithinolyticum, possibly mediated by proteinaceous cell surface-associated components, may participate in extracellular matrix degradation by activation of MMP-2 during periodontal inflammation.

Animals↗

Biofilm formation by the periodontopathic bacteria Treponema denticola and Porphyromonas gingivalis.

BACKGROUND: Periodontitis develops as a result of the interaction of the host with subgingival plaque bacteria. Both Porphyromonas gingivalis and Treponema denticola are frequently associated together in these oral biofilms. METHODS: The molecular basis for in vitro biofilm formation was investigated for P. gingivalis 381, T. denticola 35405, and mixtures of the two organisms using microtiter plate assays. In addition, the biofilms were examined following confocal laser scanning microscopy. RESULTS: P. gingivalis 381, but not T. denticola strains, formed biofilms in vitro. This property was dependent, in part, on the strain 381 fimA, ppk, and usp genes. Microarray and Northern blot analyses suggested that the expression of the ppk gene was required for maximal expression of the uspA gene. P. gingivalis 381 formed synergistic biofilms when incubated with T. denticola strains. This process was dependent upon the strain 381 rgpB and fimA genes as well as the T. denticola flgE and cfpA genes. CONCLUSIONS: P. gingivalis 381 formed synergistic biofilms with T. denticola 35405. These results may be relevant to the previous observations that the two organisms are frequently observed together in subgingival plaque with the spirochetes localized to the exterior of the oral biofilms. It is suggested that other such synergistic effects may also occur between other plaque bacteria.

Bacterial Proteins↗

Significance of detection of Porphyromonas gingivalis, Bacteroides forsythus and Treponema denticola in periodontal pockets.

The relationship between the detection of Porphyromonas gingivalis, Bacteroides forsythus and Treponema denticola in subgingival plaque samples of periodontal pockets and periodontal status was evaluated using the polymerase chain reaction (PCR). A total of 165 sites in 60 periodontitis patients were examined, and the relationships between the detection of each of the three bacterial species and the pocket depth and bleeding on probing (BOP) were analyzed. The detection ratios of P. gingivalis, B. forsythus, and T. denticola in samples from adult periodontitis lesions were 75.5%, 69.8%, and 72.6%, respectively. It was found that all sites where all three microorganisms were detected were BOP positive and had greater pocket depths than those where only one or two species were found. The detection rate of B. forsythus and T. denticola decreased with age in the sites in which PD was less than 4 mm. The present study indicates that detection of a mixed infection by P. gingivalis, B. forsythus, and T. denticola strongly correlated with adult periodontitis.

Adult↗

Effects of a mixed infection with Porphyromonas gingivalis and Treponema denticola on abscess formation and immune responses in mice.

Porphyromonas gingivalis and Treponema denticola have been found together in lesions of human periodontitis. We examined the ability of a mixed infection by both bacteria to synergistically form abscesses and disturb immune responses in mice. Absorbance of an invasive P. gingivalis 16-1 strain grown in tryptic soy broth and T. denticola ATCC 33520 strain grown in TYGVS medium were adjusted. BALB/c mice were injected with 200 microliters of the cell suspension at a site on the lateral dorsal area. The sizes of the subsequent subcutaneous abscesses were measured with a caliper gauge, and the area was expressed in square mm. Mixed infections by P. gingivalis and T.denticola produced larger abscesses than those formed after mono-infections by either P. gingivalis or T.denticola. The abscesses caused by mixed infection reached their maxima on the 6th day and maintained that size for the subsequent 5 days. The delayed type hypersensitivities against extracted antigens of P.gingivalis in mixed infection mice were significantly lower than those in the mono-infected mice. However, the IgG response to sonicated antigen of P.gingivalis did not differ between the two groups. The sizes of the abscesses caused by mixed infections in mice immunized with whole cells of P.gingivalis 16-1 were compared to those caused in sham-immunized mice. The average size of the abscess caused by mixed infection in immunized mice did not differ from that in sham-immunized mice, but many of the abscesses in immunized mice ruptured on the 4th or 5th day, followed by recovery in two weeks. These results suggest that mixed infection with P.gingivalis and T.denticola attenuates protective immune responses.

Abscess↗

Scintigraphic diagnosis of syphilitic lesions in rabbits by radiolabelled monoclonal antibodies specific for Treponema pallidum.

Immunoscintigraphy with radiolabelled monoclonal antibodies has been widely used to detect solid tumours. The purpose of this study was to investigate its potential for the specific localization of syphilitic lesions. F(ab')2 fragments were prepared from murine monoclonal antibodies against Treponema pallidum produced in our laboratory and labelled with 131I. Bilateral testicular infections were created in rabbits by inoculation with T. pallidum or Staphylococcus aureus. Seven to 10 days after inoculation, radiolabelled antibodies were injected intravenously. Serial gamma images were then taken 2 h after the injection and at 24 h intervals thereafter. Beginning as early as 2 h post-injection, the testicles could be visualized with either specific or non-specific antibodies. Gamma images in the monoclonal antibody-treated, T. pallidum-infected group persisted up to 48 h post-injection. Lesions were not discernible from background in the S. aureus-infected group injected with the monoclonal antibody and the S. aureus-infected and T. pallidum-infected groups injected with the polyclonal antibody at 24 h post-injection or later. Therefore, due to its ability to differentiate between specific and non-specific antibody-generated images from 24 h post-injection, immunoscintigraphy using monoclonal antibodies specific for T. pallidum may be employed as one of the methods to diagnose difficult cases of syphilitic internal organ involvement as well as syphilis infection in seronegative HIV-infected patients.

Animals↗

ELISA inhibition test using monoclonal antibody specific for Treponema pallidum as the serologic test for syphilis.

Monoclonal antibodies (Mabs) specific for Treponema pallidum were produced using hybridoma technology. In this study an ELISA inhibition test based on competitive inhibition by antibodies from human test sera of the binding of the enzyme-labelled Mabs to T. pallidum was performed. Inhibition of the seroreactivity of Mabs was decreased according to the dilution of rabbit antiserum to T. pallidum. Seropositivity was found in 100% of secondary and early latent syphilis patients, 75% of primary syphilis patients, and 80% of late latent syphilis patients. The mean percentage inhibition was significantly higher in each syphilis group than in the controls, and statistically significant differences were shown between all the syphilis groups. VDRL and TPHA titers were correlated with the percentage inhibitions. Therefore the ELISA inhibition test using Mab specific for T. pallidum might well be a suitable tool as a new serologic test for syphilis.

Analysis of Variance↗

Prevalence of antibodies against hepatitis A virus, hepatitis B virus, and Treponema pallidum in Mauritius.

A seroepidemiological study on the prevalence of antibodies against hepatitis A virus (HAV), hepatitis B virus (HBV) and Treponema pallidum was conducted in various groups of the population of the state of Mauritus (Islands of Mauritus and Rodrigues). 618 sera were tested. The overall prevalence of anti-HAV was 86.1% and yielded and age-dependent increase. Serological evidence for acute or chronic HBV infection was found in 3.8%; 4.5% were positive for anti-HBc alone, and in 12.6% past HBV infection was detected. No age- or sex-dependent increase in the prevalence of anti-HBc was found. There were differences in the anti-HBc prevalence among the various groups of population ranging from 5.9 (flight personnel) to 58.3% (prison inmates). Treponemal antibodies were detected in 6.0% and showed a fairly marked age-dependent increase. Our study suggests that vaccination programmes against HAV and HBV would be beneficial for the Mauritian population.

Adolescent↗

Molecular characterization of the Treponema denticola fliQ region.

A Treponema denticola 4.2 kb DNA region containing four complete genes (orfl, fliQ, fliR, and flhB) and a truncated gene (flhA') was sequenced and analyzed. The deduced amino acid sequences of FliQ, FliR, FlhB and FlhA' have significant homology with bacterial proteins associated with the flagellar export apparatus, whereas the deduced amino acid sequence of Orf1 has homology with an E. coli alcohol dehydrogenase. A putative sigma70-like promoter was identified upstream of fliQ. RT-PCR analysis indicated that fliQ, fliR, flhB and flhA' are co-transcribed independently of orfl, suggesting that the motility-associated genes are components of an operon. The location of the T. denticola fliQ-flhA' genes differs from that of the corresponding T. pallidum and Borrelia burgdorferi genes which are present in the large fla or flgB flagellar operons, respectively.

Bacterial Proteins↗

Identification and sequences of the Treponema pallidum mglA and mglC genes.

Treponema pallidum, the agent of syphilis, cannot be continuously cultivated in vitro. To identify treponemal genes encoding exported proteins, we performed TnphoA mutagenesis of a T. pallidum genomic DNA library in Escherichia coli. Clone 6D2 was chosen for further study based on partial nucleotide sequence obtained from p6D2 containing a TnphoA insertion. A complete open reading frame (orf1) and a truncated orf (orf2) were identified in the treponemal DNA of p6D2. Orf1 encodes a hydrophobic protein of 531 amino acids with a calculated M(r) of 57,882 Da. The deduced amino acid sequence of Orf1 has homology to the MglC proteins of E. coli, Haemophilus influenzae, and Salmonella typhimurium. T. pallidum Orf1 (MglC) contains a conserved motif that is found in integral cytoplasmic membrane proteins of ATP-binding cassette (ABC) transport systems. T. pallidum orf2 encodes a protein of 496 amino acids with a calculated M(r) of 55,547 Da. The deduced amino acid sequence of Orf2 has homology to the MglA proteins of S. typhimurium, E. coli, H. influenzae, and Mycoplasma genitalium. Orf2 (MglA) contains two consensus ATP-binding motifs. T. pallidum mglA and mglC are located downstream of mglB, consistent with the gene order of previously identified mgl operons. The putative T. pallidum mgl operon encodes the first high-affinity ABC transport system identified in this spirochete.

Amino Acid Sequence↗

Identification and sequences of the Treponema pallidum flhA, flhF, and orf304 genes.

The recently identified fla operon of Treponema pallidum contains several genes that encode motility-related proteins. We have determined the nucleotide sequences of three genes, designated flhA, flhF, and orf304, that are located immediately downstream of the flhB gene in the fla operon. The flhA gene encodes a 707-amino acid protein that contains five putative membrane spanning domains. FlhA has strong homology with members of a family of proteins that are involved in flagellar biogenesis and regulation/secretion of virulence-related proteins. The flhF gene encodes a 437-amino acid protein that contains three consensus elements that are characteristic of a GTP-binding domain. The orf304 gene encodes a 304-amino acid protein that contains a consensus ATP-binding motif. The order of the flhA, flhF, and orf304 genes is identical to that of corresponding genes in the Bacillus subtilis che/fla operon. Due to the location of the flhA, flhF and orf304 genes in the T. pallidum fla operon, we hypothesize that the FlhA, FlhF, and Orf304 proteins are involved in the biogenesis/assembly of treponemal periplasmic flagella.

Amino Acid Sequence↗

Nucleotide sequences of the proA and proB genes of Treponema pallidum, the syphilis agent.

The nucleotide sequences of the putative proA and proB genes of Treponema pallidum were determined. The proA gene is 1287 nucleotides long and encodes a 428 amino acid protein with a predicted M(r) of 46.6 kDa. The proB gene is 891 nucleotides long and encodes a 296 amino acid protein with a predicted M(r) of 31.3 kDa. The deduced amino acid sequences of the treponemal ProA and ProB proteins have a high degree of homology to the amino acid sequences of several bacterial ProA and ProB proteins. The order of the T. pallidum pro genes (proA/proB) is unique in comparison to the order of these genes in other bacteria. The identification of the putative proA and proB genes in T. pallidum, coupled with the previous identification of the proC gene, strongly suggests that this fastidious spirochete is capable of proline biosynthesis.

Aldehyde Oxidoreductases↗