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N-terminal amino acid sequence of the Borrelia burgdorferi flagellin.

The 41 kDa flagellar protein of Borrelia burgdorferi appears to be an immunodominant antigen producing an early and strong response in most, if not all, individuals during infection in humans. It would represent a very good antigen for serodiagnosis of Lyme disease, if its crossreactivity with flagella of other bacteria was low. To gain information on this point we isolated the B. burgdorferi flagellin by preparative two-dimensional electrophoresis for N-terminal amino acid analysis. By comparing the N-terminal amino acid sequences of flagellar proteins from other eubacteria we found that the first six out of twenty nine amino acids were identical to the Treponema pallidum and Treponema phagedenis 'class B' flagellins. All 29 N-terminal residues exhibited a moderate inter-genus homology (44-55%), in contrast to the high degree (67-95%) of inter-species conservation of the treponemal 'class B' flagellar N-terminal sequences. There was little similarity to other flagellins except the B. subtilis flagellar protein.

Amino Acid Sequence↗

Sexual transmission of hepatitis B in Mwanza, Tanzania.

BACKGROUND: Hepatitis B virus (HBV) is endemic and poses a grave public health problem in Africa where it is mainly transmitted from mother to baby or during childhood. Sexual transmission has also been suggested to play a role in East Africa, but this has never been properly demonstrated. Additional preventive strategies may be proposed if sexual transmission of HBV occurred in this region where HIV and other STDs are highly prevalent. GOALS: To determine the prevalence of markers for hepatitis B virus (HBV)and other sexually transmitted diseases (STD) in routine blood samples taken from three populations in Mwanza, Tanzania, and to use the data collected to look at the association between hepatitis B and other STDs, including human immunodeficiency virus (HIV). STUDY DESIGN: Routine blood samples were collected from 1,025 patients attending a clinic for STDs, 253 voluntary blood donors from secondary schools, and 952 blood donors who gave blood in a hospital specifically for a relative who needed a blood transfusion. All samples were tested for HIV by double enzyme-linked immunosorbent assay (ELISA), and for syphilis using the Treponema pallidum hemagglutination (TPHA) and rapid plasma reagin (RPR) tests. Two markers for HBV were examined by the double ELISA method, the presence of the anti-hepatitis B core antigen (anti-HBc) and the hepatitis B surface antigen (HBsAg). RESULTS: There were high prevalences of HBV, syphilis, and HIV in relative donors and STD patients. Although HBV markers were more prevalent in men of increasing ages, syphilis and HIV markers were more prevalent in young women. Evidence of past infection with HBV (presence of anti-HBc) was associated with serologic markers of recent treponemal infection (both TPHA and RPR positive) in both sexes (men odds ratio [OR] = 1.91, P < 0.011; women OR = 2.34, P < 0.02) and with HIV in men (OR = 1.93, P < 0.003). Current infection with HBV (presence of HBsAg) was associated with recent syphilis in men (OR = 2.13, P < 0.006). In STD patients, current infection with HBV was associated with Trichomonas vaginalis in women (OR = 3.57, P < 0.002) and recent syphilis in men (OR = 3.46, P < 0.001). There was no further association between HBV markers and any other STD pathogen or any particular STD syndrome, nor was there any association between current HBV infection and HIV in both sexes. The population attributable fraction for sexual acquisition of hepatitis B is estimated at 7.2% in men and 3.0% in women, based on the association between hepatitis B and syphilis. CONCLUSIONS: These findings suggest that sexual acquisition of hepatitis B occurs at low levels in Mwanza, and that HBV can be prevented through enhancement of the current HIV/STD control activities, in addition to improved vaccination strategies.

Adolescent↗

Tiamulin water medication in the treatment of swine dysentery under farm conditions.

The clinical efficacy of tiamulin given in the drinking water at levels of 0.0045 per cent for five days and 0.006 per cent for three days was evaluated for the treatment of swine dysentery in six clinical studies and compared with positive controls using tylosin at 0.02 per cent in the drinking water for five days and untreated controls. Efficacy was evaluated by assessment of the pigs' general appearance; the extent of diarrhoea and dehydration; performance, as measured by average daily gains and feed conversion efficiency; the presence or absence of Treponema hyodysenteriae in the faeces and mortality. As a result of statistical analysis of these parameters tiamulin medication was found to be an efficacious therapeutic regimen for naturally occurring swine dysentery when given at either level in the drinking water.

Animals↗

Characterization of the 35-kilodalton Treponema pallidum subsp. pallidum recombinant lipoprotein TmpC and antibody response to lipidated and nonlipidated T. pallidum antigens.

The gene encoding the 35-kDa immunogenic Treponema pallidium subsp. pallidum (T. pallidum) membrane protein C, TmpC, was cloned, sequenced, and expressed in Escherichia coli. The deduced amino acid sequence carries on N-terminal signal sequence with a four-amino-acid motif, which is characteristic for bacterial lipoproteins. Metabolic labeling with radioactive palmitic acid of E. coli expressing TmpC revealed incorporation of the fatty acid into the antigen. The antigen was overproduced, purified to near homogeneity and used in an enzyme-linked immunosorbent assay (ELISA) to evaluate its potential for the serodiagnosis of syphilis. Although all sera from untreated secondary syphilis patients were reactive in this TmpC ELISA, only a minority of the serum samples from untreated patients in the primary or early latent stage of the disease contained significant anti-TmpC antibodies. To study the influence of the lipid moiety on the antigenic properties of the TmpC, TmpA, and TpD lipoproteins, plasmids encoding nonlipidated forms of these antigens were constructed. In addition, a plasmid expressing a lipidated form of the otherwise non-lipid-modified antigen TmpB was constructed. Immunization and absorption experiments with these lipidated and nonlipidated antigens showed that antibodies against the lipid moiety of lipoproteins could not be detected on immunoblots, neither in sera from infected rabbits nor in sera from animals immunized with the lipoproteins. In addition, we were unable to demonstrate cross-reactivity between antibodies against the T. pallidum lipoproteins and those reactive to the Venereal Diseases Research Laboratories test, suggesting that antibodies reactive to the Venereal Diseases Research Laboratories test are unrelated to antilipoprotein antibodies.

Amino Acid Sequence↗

The specificity of fetal IgM: antibody or anti-antibody?

Reference materials were produced to standardize the immunoglobulin class specificity and potency of immunofluorescent anti-IgM conjugates used for diagnostic tests for congenital syphilis. In attempting to mimic essential immunologic characteristics of syphilitic and nonsyphilitic infant sera, we evaluated these sera in comparison with processed adult sera. We were quite surprised to discover that some syphilitic babies do not produce significant quantities to IgM antibody to T. pallidum in response to their infection, as would be expected; instead, they make relatively large amounts of IgM anti-IgG. We found this to be true also for newborns and infants infected with cytomegalovirus, rubella, and toxoplasmosis. To our knowledge, this observation has not been previously reported. However, it could have been predicted from the knowledge that older infants and young children normally produce IgM antibodies to maternal IgG allotypes (Gm factors). We are disturbed that these findings suggest that currently recommended indirect immunofluorescence IgM tests for perinatal infection may not be disease specific. Our observations may be important for a better understanding of basic immunologic mechanisms of fetal-maternal to tolerance and fetal response to life-threatening infection.

Adsorption↗

Two cases of acquired syphilis with acute central chorioretinitis as initial manifestation.

Acquired syphilis has become an overlooked cause of posterior uveitis including chorioretinitis. Two male cases of bilateral acute acquired syphilitic chorioretinitis were observed, one of whom demonstrated an early delay of choroidal circulation during fluorescein angiography, possibly indicating the choroidal or subretinal neovascular proliferation which occurred later. Although both patients had been treated with oral prednisone for approximately one month before their initial visit to our clinic, good visual recovery had not been achieved. Diagnosis was first confirmed by the positive results of serologic Treponema pallidum hemagglutination and Venereal Disease Research Laboratory tests. The patients were successfully treated with orally administered bacampicillin, which resulted in the complete recovery of visual acuity and normal fundus appearance except for slight pigmentary changes of the retina in the macular area.

Acute Disease↗

Oral flora in the age of molecular biology.

The present work describes the fundamental aspects of the molecular methods that are applied to oral microbiology: probes, PCR, multiple real time PCR, 16S rDNA, and proteomics. Likewise, it presents the results obtained by applying these methods to the study of the bacterial flora encountered inside the mouth. By identifying 16S rDNA, up to 132 known species and 215 new, unknown phylotypes have been detected inside patients s periodontal pockets. We are currently using a commercial system based on multiple PCR (genomic amplification), followed by reverse hybridization to detect the five species known to cause periodontal disease. We also summarize the findings derived from the application of proteomic techniques to the study of odontologic infections pathogenesis and from the use of molecular methods in the study of resistance to antimicrobial agents. Finally, it puts forth the problems that remain unsolved with respect to oral flora and the treatment of odontogenic infections. Traditional culture methods continue to be indispensable, as they make it possible to later work with the cultured microorganisms.

Aggregatibacter actinomycetemcomitans↗

Serological tests for syphilis in Saudi Arabia.

A total of 6684 sera were initially screened for syphilis by the Venereal Disease Research Laboratory (VDRL) test and the Treponema pallidum haemagglutination assay (TPHA). Reactive sera from either or both these tests were tested for confirmation by the fluorescent treponemal antibody-absorbed (FTA-ABS) test. VDRL biological false positive reactors were detected in 0.5% of the total sera examined, with 0.4% and 0.8%, respectively, obtained in pregnant women and blood donors. Eight sera (0.1%) were found to be positive in the TPHA test alone. An overall positivity of 2.7% for syphilis was detected, with a 0.85% positivity in antenatal patients. Infection with T pallidum seemed to be more common in men than in women (1.6:1) and predominated in the age group 20-39 years. Serological testing of sera from 26 mother and infant pairs allowed one case of congenital syphilis to be detected by FTA-ABS (IgM) and identified VDRL biological false positivity in seven infants.

Adult↗

Identification of oral spirochetes at the species level and their association with other bacteria in endodontic infections.

OBJECTIVES: Recent molecular approaches have revealed that fastidious organisms such as Bacteroides forsythus and oral treponemes were frequently found in root canals with apical periodontitis. The purpose of this study was to identify the isolates of oral spirochetes at the species level in endodontic infections and to determine their association with B forsythus and Porphyromonas gingivalis. STUDY DESIGN: Seventy-nine teeth with apical periodontitis were selected for this study. After sampling from the root canals aseptically, polymerase chain reaction amplification for the 16S rRNA gene was performed with eubacterial universal primers. Subsequently, dot-blot hybridization was performed with 8 species-specific oligonucleotide probes. The microbial associations were analyzed by using the odds ratio. RESULTS: The most frequently found species was P gingivalis (27.4%), followed by Treponema maltophilum (26%), B forsythus (16.4%), and Treponema socranskii (2.7%). Other treponemes, including Treponema denticola, were not detected in our samples. Significant microbial associations were identified between T maltophilum, B forsythus, and P gingivalis by performing analysis with the odds ratio. CONCLUSION: Our results indicate that T maltophilum should be included in etiologic studies of endodontic diseases.

Adolescent↗

The role of Neisseria gonorrhoeae and Chlamydia trachomatis in pelvic inflammatory disease and its sequelae in Zimbabwe.

The presence of antibodies to pili of Neisseria gonorrhoeae and Chlamydia trachomatis serovar L2 were assessed in women consecutively hospitalized in Zimbabwe with pelvic inflammatory disease (PID; n = 66), infertility (n = 227), and ectopic pregnancy (n = 60). Women delivering live full-term infants served as controls. Of the infertile women, 60% had secondary infertility; 59% had macroscopic evidence of a tubal abnormality. Women with PID, infertility and tubal disease, and ectopic pregnancy and tubal disease had significantly higher prevalences of antibodies against C. trachomatis and N. gonorrhoeae than did controls or women with infertility or ectopic pregnancy but no macroscopic tubal abnormalities (P less than .001 for all comparisons). The prevalence of antibody to chlamydia increased with age (P = .01), unlike the gonococcal antibody. Antibodies to C. trachomatis were associated with a history of PID, being single, a positive Treponema pallidum hemagglutination assay, and chlamydial antibody. None of the controls had human immunodeficiency virus, unlike 3.9%-7.6% of the other women. Tubal abnormalities were implicated in more than half of the cases of infertility.

Adult↗

The microbiological case for adjunctive therapy for periodontitis.

That chronic periodontitis is an infectious disease is now firmly established, and the primary role of Porphyromonas gingivalis, Tannerella forsythensis and Treponema denticola is generally accepted. Treatment by mechanical means such as scaling and root planing or surgery generally results in significant clinical improvement but may not arrest the progress or recurrence of disease. Several studies have shown that the probability of achieving lasting stability as measured by the arrest of progressive attachment loss and bone loss by primary mechanical therapy is a function, in major part, of whether pathogenic microorganisms are still present at local subgingival sites at the completion of active therapy. The infecting bacterial species are susceptible to killing by several antibiotics including, among others, tetracycline-class drugs, amoxicillin and metronidazole as well as by local exposure to chlorhexidine. Randomized clinical trials have shown that use of systemically administered antibiotics as an adjunct to mechanical therapies significantly enhances clinical outcomes and stability. Several slow-release devices that deliver anti-microbial drugs directly into periodontal pockets have been developed and are now on the market. Use of these devices permits local delivery of long-lasting, high concentrations of doxycycline (Atridox) minocycline (Arestin), and chlorhexidine (PerioChip) directly into periodontal pockets. Although these devices differ with regard to ease of use, concentration of drug delivered and length of time high drug concentrations can be maintained, randomized clinical trials have shown that their use as an adjunctive treatment to scaling and root planing results in a significantly greater reduction of periodontal pocket depth and an average increase in clinical periodontal attachment level of about 0.8 mm. Gain in clinical attachment is greater in deeper pockets than in shallower pockets. Locally delivered adjunctive anti-microbial therapy is an effective means to enhance therapeutic outcomes.

Anti-Bacterial Agents↗

Is antenatal screening for syphilis worth while?

To ascertain the effectiveness of an antenatal screening programme for syphilis, a review of the seven-year period from 1981 to 1987, was made at The Royal Women's Hospital, Melbourne. In total, 35,103 patients were screened, with a biological false-positive rate of 0.3%. Serological evidence of treponemal infection occurred in 0.4% of patients, with 66% of this group being newly recognized cases. Eight (0.02%) cases were consistent serologically with early treponemal infection, although clinically all patients were asymptomatic at the time of detection. The outcome in infants was satisfactory in all cases where appropriate antibiotic therapy was instituted antenatally. In one case, which presented late at 32-weeks' gestation for the first antenatal visit, the result was a fetal death in utero. The post-mortem findings were compatible with a diagnosis of congenital syphilis. At an incidence of 0.02% for active maternal syphilis, and an over-all treponemal infection rate of 0.4%, the cost-effectiveness of antenatal screening is well-justified.

Cost-Benefit Analysis↗

Syphilis.

In the last half of the 20(th) century, medicine has made tremendous inroads against syphilis chiefly owing to the introduction of penicillin and vigorous public health initiatives. Yet, the world continues to be burdened by this disease. Since 2000, overall rates of syphilis have risen in the US and throughout the world. Furthermore, through its association with an increased risk of HIV infection, syphilis has acquired a new potential for morbidity and mortality. The aim of this review is to survey the latest knowledge about syphilis, including pathogenesis and host response, clinical manifestations, diagnostic methods and, especially, treatment and follow-up.

Algorithms↗

Cofactors for the acquisition of HIV-1 among heterosexual men: prospective cohort study of trucking company workers in Kenya.

OBJECTIVE: To determine the prevalence, incidence, and correlates of HIV-1 infection in a cohort of east African trucking company employees. METHODS: HIV-1-seronegative trucking company employees were enrolled in a prospective cohort study and evaluated at 3 monthly intervals for HIV-1 seroconversion, sexually transmitted diseases, and sexual behavior. RESULTS: The baseline seroprevalence of HIV-1 among 1500 trucking company employees was 17.8%. Among 752 HIV-1-seronegative men who were followed, the HIV-1 annual seroincidence was 3.1%. In univariate analysis, HIV-1 acquisition was associated with age under 25 years, 10 years or less of sexual activity, occupation as a driver/driver's assistant, occupational travel for more than 14 days per month, religion other than Christian or Muslim, uncircumcised status, sex with a prostitute, sex with a girlfriend/casual partner, extramarital sex, and enrollment seropositivity to Treponema pallidum, Haemophilus ducreyi, and Herpes simplex virus type 2 (all P values < or = 0.05). Using multivariate analysis, HIV-1 acquisition was independently associated with 10 years or less of sexual activity (hazard rate ratio (HRR) 2.0, 95% confidence interval (CI) 1.0-4.3), occupation as a driver/driver's assistant (HRR 3.9, 95% CI 1.7-9.0), religion other than Christian or Muslim (HRR 6.1, 95% CI 1.4-25.7), uncircumcised status (HRR 2.3, 95% CI 1.0-5.0), and unprotected sex with a prostitute (HRR 2.8, 95% CI 1.1-7.0). CONCLUSIONS: Trucking company employees had a high HIV-1 seroprevalence rate at enrollment and a high HIV-1 seroincidence during follow-up. Risk factors for HIV-1 seroconversion included years of sexual activity, occupation, religion, uncircumcised status, and unprotected sex with a prostitute. This population is an appropriate target for HIV-1 prevention trials and behavioral interventions.

Adolescent↗

Evidence for Lyme disease in urban park workers: a potential new health hazard for city inhabitants.

In the UK, cases of Lyme disease have only been reported from rural areas. Recently, however, Ixodes ticks infected with Borrelia burgdorferi have been found in London parks. To determine whether this constituted a health hazard, we questioned 44 workers from Richmond and Bushey parks to assess their exposure to tick bites and whether they had a clinical history of Lyme disease. Their serum was subsequently investigated for antibodies to two different preparations of Borrelia burgdorferi (whole cell sonicate and flagellin) and the specificity of these antibodies determined by immunoblotting. Comparison was made to zoo keepers (n = 27) from a wildlife park outside London. Tick bites were reported in 23% of park workers and of these, three described symptoms compatible with Lyme disease. Raised antibody levels were found in 10 (24%) of the park workers compared with one (4%) of the zoo keepers using ELISA with whole cell sonicate as antigen (P = 0.02) and 6 (14%) of park workers and none of the zoo keepers using purified flagellin as antigen (P < 0.05). Analysis of the immunoblots revealed more bands were detected in park workers (mean 1.8, range 0-6) than in the zoo keeper controls (mean 0.8, range 0-4); P < 0.001 and 14 (32%) of the park workers had reactivity with three or more protein bands, whilst only one of the zoo keepers showed this level of antigen binding (P < 0.005). These data suggest previous infection with B. burgdorferi in London park workers which has important health implications for these individuals, other park workers and possibly park visitors.

Adolescent↗

Necrotizing ulcerative gingivitis, periodontitis, and stomatitis: clinical staging and predisposing factors.

Necrotizing ulcerative gingivitis (NUG), necrotizing ulcerative periodontitis (NUP), and necrotizing stomatitis (NS), collectively termed necrotizing gingivostomatitis (NG), represent a dramatic, but rare oral infection associated with diminished systemic resistance, including HIV infection. Over a 5-year period, 68 consecutive NG patients from a population with known HIV status were evaluated and treated. Lesions were staged (modified Pindborg), and clinical findings and predictor variables were compared to 68 random control subjects without NG. Most cases (52%) were stage 1, with necrosis of the tip of the interdental papilla only; 19% were stage 2, with the entire papilla affected; 22% had necrosis of marginal (stage 3) or attached gingiva (stage 4); and 7% were more advanced, with mucosal necrosis or bone exposure. Attachment loss was a feature of stage 2 or greater NG. Beside HIV infection, significant predisposing factors included poor oral hygiene, unusual life stress, inadequate sleep, Caucasian race, age 18 to 21 years, and recent illness. Ten of 68 NG patients were HIV-positive. These patients were older than seronegative patients, less likely to be Caucasian, and maintained better oral hygiene and sleep. HIV-positive NG cases were clinically indistinguishable from HIV-negative cases in this series.

Adult↗

Efficacy of aztreonam in treatment of experimental syphilis in rabbits.

The efficacy of aztreonam (SQ 26,776) in the therapy of active syphilis infection was evaluated in the rabbit model. Aztreonam was effective in treating active syphilis at a dose of 25 mg/kg given intramuscularly twice daily for 10 days; doses of 2.5 and 0.25 mg/kg were not effective.

Animals↗

A new technique of heterogenous enzyme-linked immunosorbent assay, stick-ELISA. II. Application of stick-ELISA to antigens of various infective agents.

Stick-enzyme-linked immunosorbent assay (Stick-ELISA) was applied to antigens of a variety of infective agents. An acceptable reproducibility in a series was reached, the variation coefficient being twelve percent. Preliminary results revealed that ready coated sticks could be stored at 4 degrees C for at least two to three weeks.

Antigens↗