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PubMed1989

Demonstration of rare protein in the outer membrane of Treponema pallidum subsp. pallidum by freeze-fracture analysis.

The surface of Treponema pallidum subsp. pallidum (T. pallidum), the etiologic agent of syphilis, appears antigenically inert and lacks detectable protein, as judged by immunocytochemical and biochemical techniques commonly used to identify the outer membrane (OM) constituents of gram-negative bacteria. We examined T. pallidum by freeze-fracture electron microscopy to visualize the architecture of its OM. Treponema phagedenis biotype Reiter (T. phagedenis Reiter), a nonpathogenic host-associated treponeme, and Spirochaeta aurantia, a free-living spirochete, were studied similarly. Few intramembranous particles interrupted the smooth convex and concave fracture faces of the OM of T. pallidum, demonstrating that the OM of this organism is an unusual, nearly naked lipid bilayer. In contrast, the concave fracture face of the OM of S. aurantia was densely covered with particles, indicating the presence of abundant integral membrane proteins, a feature shared by typical gram-negative organisms. The concentration of particles in the OM concave fracture face of T. phagedenis Reiter was intermediate between those of T. pallidum and S. aurantia. Similar to typical gram-negative bacteria, the OM convex fracture faces of the three spirochetes contained relatively few particles. The unique molecular architecture of the OM of T. pallidum can explain the puzzling in vitro properties of the surface of the organism and may reflect a specific adaptation by which treponemes evade the host immune response.

Bacterial Outer Membrane Proteins↗

PubMed1996

Characterization of the cytoplasmic filament protein gene (cfpA) of Treponema pallidum subsp. pallidum.

Treponema pallidum and other members of the genera Treponema, Spirochaeta, and Leptonema contain multiple cytoplasmic filaments that run the length of the organism just underneath the cytoplasmic membrane. These cytoplasmic filaments have a ribbon-like profile and consist of a major cytoplasmic filament protein subunit (CfpA, formerly called TpN83) with a relative molecular weight of approximately 80,000. Degenerate DNA primers based on N-terminal and CNBr cleavage fragment amino acid sequences of T. pallidum subsp. pallidum (Nichols) CfpA were utilized to amplify a fragment of the encoding gene (cfpA). A 6.8-kb EcoRI fragment containing all but the 5' end of cfpA was identified by hybridization with the resulting PCR product and cloned into Lambda ZAP II. The 5' region was obtained by inverse PCR, and the complete gene sequence was determined. The cfpA sequence contained a 2,034-nucleotide coding region, a putative promoter with consensus sequences (5'-TTTACA-3' for -35 and 5'-TACAAT-3' for -10) similar to the sigma70 recognition sequence of Escherichia coli and other organisms, and a putative ribosome-binding site (5'-AGGAG-3'). The deduced amino acid sequence of CfpA indicated a protein of 678 residues with a calculated molecular mass of 78.5 kDa and an estimated pI of 6.15. No significant homology to known proteins or structural motifs was found among known prokaryotic or eukaryotic sequences. Expression of a LacZ-CfpA fusion protein in E. coli was detrimental to survival and growth of the host strain and resulted in the formation of short, irregular filaments suggestive of partial self-assembly of CfpA. The cytoplasmic filaments of T. pallidum and other spirochetes appear to represent a unique form of prokaryotic intracytoplasmic inclusions.

Amino Acid Sequence↗

PubMed1998

Chemotaxis in Borrelia burgdorferi.

Borrelia burgdorferi is a motile spirochete which has been identified as the causative microorganism in Lyme disease. The physiological functions which govern the motility of this organism have not been elucidated. In this study, we found that motility of B. burgdorferi required an environment similar to interstitial fluid (e.g., pH 7.6 and 0.15 M NaCl). Several methods were used to detect and measure chemotaxis of B. burgdorferi. A number of chemical compounds and mixtures were surveyed for the ability to induce positive and negative chemotaxis of B. burgdorferi. Rabbit serum was found to be an attractant for B. burgdorferi, while ethanol and butanol were found to be repellents. Unlike some free-living spirochetes (e.g., Spirochaeta aurantia), B. burgdorferi did not exhibit any observable chemotaxis to common sugars or amino acids. A method was developed to produce spirochete cells with a self-entangled end. These cells enabled us to study the rotation of a single flagellar bundle in response to chemoattractants or repellents. The study shows that the frequency and duration for pausing of flagella are important for chemotaxis of B. burgdorferi.

Adaptation, Physiological↗

PubMed2000

The spirochete FlaA periplasmic flagellar sheath protein impacts flagellar helicity.

Spirochete periplasmic flagella (PFs), including those from Brachyspira (Serpulina), Spirochaeta, Treponema, and Leptospira spp., have a unique structure. In most spirochete species, the periplasmic flagellar filaments consist of a core of at least three proteins (FlaB1, FlaB2, and FlaB3) and a sheath protein (FlaA). Each of these proteins is encoded by a separate gene. Using Brachyspira hyodysenteriae as a model system for analyzing PF function by allelic exchange mutagenesis, we analyzed purified PFs from previously constructed flaA::cat, flaA::kan, and flaB1::kan mutants and newly constructed flaB2::cat and flaB3::cat mutants. We investigated whether any of these mutants had a loss of motility and altered PF structure. As formerly found with flaA::cat, flaA::kan, and flaB1::kan mutants, flaB2::cat and flaB3::cat mutants were still motile, but all were less motile than the wild-type strain, using a swarm-plate assay. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blot analysis indicated that each mutation resulted in the specific loss of the cognate gene product in the assembled purified PFs. Consistent with these results, Northern blot analysis indicated that each flagellar filament gene was monocistronic. In contrast to previous results that analyzed PFs attached to disrupted cells, purified PFs from a flaA::cat mutant were significantly thinner (19.6 nm) than those of the wild-type strain and flaB1::kan, flaB2::cat, and flaB3::cat mutants (24 to 25 nm). These results provide supportive genetic evidence that FlaA forms a sheath around the FlaB core. Using high-magnification dark-field microscopy, we also found that flaA::cat and flaA::kan mutants produced PFs with a smaller helix pitch and helix diameter compared to the wild-type strain and flaB mutants. These results indicate that the interaction of FlaA with the FlaB core impacts periplasmic flagellar helical morphology.

Alleles↗

PubMed1987

New infectious spirochete isolated from short-tailed shrews and white-footed mice.

A spirochete with two periplasmic flagella was isolated from the blood or tissues of spleens and kidneys from short-tailed shrews (Blarina brevicauda) and white-footed mice (Peromyscus leucopus) in Connecticut and Minnesota. After inoculation, the shrew-mouse spirochete infected Swiss mice and Syrian hamsters. This spirochete is morphologically and serologically distinct from the species of Treponema, Borrelia, Leptospira, and Spirochaeta examined.

Animals↗

PubMed1992

Polymerase chain reaction for detection of Leptospira spp. in clinical samples.

A sensitive assay for Leptospira spp., the causative agent of leptospirosis, was developed on the basis of the polymerase chain reaction (PCR). A 331-bp sequence from the Leptospira interrogans serovar canicola rrs (16S) gene was amplified, and the PCR products were analyzed by DNA-DNA hybridization by using a 289-bp fragment internal to the amplified DNA. Specific PCR products also were obtained with DNA from the closely related nonpathogenic Leptospira biflexa but not with DNA from other spirochetes, such as Borrelia burgdorferi, Borrelia hermsii, Treponema denticola, Treponema pallidum, Spirochaeta aurantia, or more distant organisms such as Escherichia coli, Staphylococcus aureus, Mycobacterium tuberculosis, and Proteus mirabilis. The assay was able to detect as few as 10 bacteria. Leptospira DNA was detected in urine from experimentally infected mice. In addition, the test was found to be suitable for diagnosing leptospirosis in humans. Cerebrospinal fluid and urine from patients with leptospirosis were positive, whereas samples from control uninfected patients were negative.

Adolescent↗

PubMed1993

[What mouse contributed the first representation of the adrenal cortex X zone?].

The X zone, innermost cortex of the mouse adrenal, was first represented by Kiyoshi Masui and Yasushige Tamura in the Japanese in 1924 and in the English in 1926. All the mice used by them were reared by themselves at the Laboratory of Veterinary Anatomy, Faculty of Agriculture, Imperial University of Tokyo. They were the first scientists in Japan that attempted to breed mice in their own laboratory and used them for scientific researches in the literature. The importance of the breeding of laboratory animals by users was first pointed out by Sahachiro Hata in 1914, who found Salvarsan as a specific medicine against Spirochaeta pallida in cooperation with Paul Ehrlich in 1910. Hata's idea on the breeding of laboratory animals by researchers themselves might inspire Masui. Hata himself tried to breed laboratory mice in the Kitasato Institute. They were called "French mice", because they were believed at that time to be introduced from Europe before 1910. Masui and Seitaro Adachi (1926) also used French mice in their basic studies. They compared the body size between French and Nanking mice with special attention to the genetic factors controlling the body size for the purpose of improving of army horses. They noticed that French mice were larger in body size and more active in reproduction. Hata noticed a difference in reaction to Salvarsan between two mice. Our survey of the literature revealed that the X zone in the mice used by Masui and Tamura had many distinctive characteristics. The major two found in 90-day-old virgins were the extremely thick X zone and its division into two sublayers, one being composed of vacuolated cells only and the other of non-vacuolated cells only. It seems most reasonable to postulate that the first representation of the X zone was made in French mice bred by Masui and Tamura themselves. These characteristics of the X zone in the mice used by Hata or Masui and Tamura were very similar to those observed in inbred strains of mice, KK and NC, which Kyoji Kondo established from Japanese fancy mice, Kasukabe and Nishikinezumi groups, respectively. The same characteristics were also found in 4 of 28 recombinant inbred (RI) strains between A/J and SM/J established by Masahiko Nishimura. Since both A/J and SM/J have not such characteristics, the thick X zone consisting of 2 sublayers with and without vacuolation seems to appear on a certain specific genetic background.(ABSTRACT TRUNCATED AT 400 WORDS)

Adrenal Cortex↗

PubMedDate not supplied

[Mouth diseases and dental materials: potential for study with cytofluorometry].

BACKGROUND: The potential dental materials have of inducing mouth diseases is well known, and various methods have been developed to investigate this phenomenon. Among these there are histological studies of the pulp and periodontium exposed to dental materials, both in vivo and in vitro in humans and animals. Other studies are based on the clinical observation of the effects induced by dental materials. Aim of the paper is to evaluate the use of flow cytometry to analyse crevicular fluid to study its content in terms of inflammatory cells and inflammation mediators. METHODS: Samples of crevicular fluid were collected by aspiration using a bevelled needle mounted on a 5 ml syringe from patients without periodontitis and with periodontitis and multiple heterogeneous dental restorations. This method was adopted since it allows to place the cells of the fluid in suspension. Part of the fluid was analysed by cytology and part by flow cytometry. RESULTS: In the patients without periodontal disease cytological examination revealed the presence of desquamated epithelial cells and colonies of cocci, rods and spirochaetae. In the patients with periodontal disease the same examination revealed, besides the above, the presence of inflammatory cells. Flow cytometry confirmed the findings of cytological examination, thus proving to be an effective method for studying crevicular fluid. CONCLUSIONS: This method allows to identify specific inflammatory cells or mediators of inflammation in crevicular fluid and may therefore prove to be very useful in the study of alterations induced by some dental materials at the level of the gingival sulcus.

Adult↗

PubMed1996

[Paul Ehrlich (1854-1915) and his contacts with East European physicians at the time of testing the first chemioterapeutics].

Among numerous scientific contacts which Paul Ehrlich had with foreign scientists information about his contacts with East European scientists is very limited. The communiqué will deal with some of the particulars concerning Ehrlich's co-operation with physicians from East Europe directly before the practical application of the chemioterapeutics among others he was in touch with the Russians: Georgi N. Gabryczewski, Alexander A. Władimirow and a Finn Julius Iversen. The contacts originated during Robert Koch bacteriological courses at the Institute of Hygiene in Berlin. Only between 1885-1988 at least 11% of participants came from the countries of Eastern Europe. Ehrlich and Gabryczewski correspondence indicates that such a co-operation had already existed in the 90-ties, at the time of the research of a therapeutic influence of aniline dyes like diacetylparaaminophenol and triasid at spirillioza (recurrens). Iversen, a chief physician of Obuchow Hospital in St. Petersburg, was one of the first who after initial tests (Atoxyl, Arsazetin), at Ehrlich's request successfully used 606 preparation in case of spirochaeta infections.

Chemistry, Pharmaceutical↗

PubMedDate not supplied

[Comparative study of the energy metabolism of anaerobic alkaliphiles from soda lakes].

We investigated the influence of inhibitors of energy metabolism and ionophores on the growth and formation of metabolic products in alkaliphilic anaerobes characterized by various catabolism types. It was shown that blockage of oxidative phosphorylation by the addition of N,N'-dicyclohexylcarbodiimide (DCCD), an inhibitor of F1F0 ATP synthase, resulted in a complete arrest of the growth of the acetogenic bacterium Tindallia magadiensis with arginine as electron acceptor. In the presence of pyruvate, substrate-level phosphorylation occurred. The methylotrophic methanogenic archaebacterium Methanosalus zhilinae did not grow with DCCD and vanadate, an inhibitor of E1E2 ATPase, suggesting the presence of two ATPase types in this species. In the saccharolytic alkaliphiles Halonatronum, Amphibacillus tropicus, and Spirochaeta alkalica (which are characterized by different pH optima), the contribution of the H+ gradient to the energy metabolism and, presumably, to the maintenance of the intracellular pH level decreased with an increase in the degree of alkaliphily. Based on the data of an inhibitor assay using protonophores, monensin, and amiloride, we suggest that all of the bacteria tested depend on H+- and Na+-gradients. The Na+/H+ antiport appears to be a universal mechanism of regulating the intracellular pH level and the interaction between the Na+ and the H+ cycles in bacterial cells cultivated under alkaline conditions.

Alkalies↗

PubMed1992

[Incidence of Borrelia burgdorferi infection in isolated facial paralysis. A prospective study].

Between the causes of facial paralysis, similar to Bell's palsy, recently are included those due to some families of spirochaeta (Borrelia burgdorferi) transmitted through tick-bite. These bacteria, besides de facial palsy, may produce several degrees of lymphocytic meningitis and subsequently the paralysis of the face, sometimes clumsy or recurrent. The treatment being the antibiotic drugs, never the corticoids. In the paper are explained prospectively the incidence of this disease in the AA's environment.

Adolescent↗

PubMed2006

[Allele variants of Borrelia afzelii found by sequencing of the chromosome gene p66].

Sequencing of gene p66 fragments of 246-337 p.b. was performed in 45 isolates of Aorrelia afzelii isolated from different transmitting agents and reservoir hosts within the habitation area of the spirochaeta. At least seven allele variants of the pathogenic agent were found indifferent natural foci of the disease. The extent of similarity between the nucleotide sequences of the isolates of the same allele variant was 99.9-100%; the extent of similarity between different allele variants was 98.9-99.7%. It was found that the majority of genovariants of A. afzelii (with respect to 5S-23S) incorporated several different allele variants of gene p66, all allele variants being found in the two genogroups (VS461 and NT28) of the pathogenic agent.

Alleles↗

PubMed1991

[Karl Landsteiner (1868-1943) in memoriam].

Apart from the discovery of the human blood groups and the Rhesus factor Landsteiner developed the visualization of Spirochaeta pallida by the method of dark field microscopy. Furthermore, he demonstrated the transmission of poliomyelitis to monkeys, which made it possible to prove the viral origin of this disease. He gave fundamental impulses to the development of immunobiology (e.g. concept of haptens, specificity of antigens etc.).

Austria↗

PubMed1990

[Isolation and identification of oral spirochaetes and Fusobacterium in juvenile periodontitis].

Periodontal pocket specimens of 40 juveniles periodontitis (JP) and specimens from gingival sulcus in 40 normal juveniles (NP) were examined. 25 strains of anaerobic oral spirochaete (62.5%) (17 strains were Treponema microdentium, 3 strains macrodentium, 2 strains T orale, 3 strains Borrelia) and 21 strains of fusobacterium were isolated in 40 JP. In 40 NP, 2 strains of spirochaetes (5%) and 3 strains fusobacterium (7.5%) were found. This study shows that there is a relationship between the infection of spirochaeta, fusobacterium and JP.

Adolescent↗

PubMed1989

[Incidence of the spirochete Borrelia burgdorferi in arthropods (Arthropoda) and antibodies in vertebrates (Vertebrata)].

The paper summarizes data on hitherto assembled findings of Spirochaeta burgdorferi, the causal agent of Lyme disease in arthropods and the incidence of antibodies in birds and mammals. The authors evaluate some vectors and reservoir animals, incl. possible carriers. Borrelia burgdorferi was found so far in 30 species of Arthropoda, 13 species of mites (Acarina), 15 species of flies(Diptera), two species of fleas (Siphonaptera). The role of insects as vectors was not proved so far. Antibodies were detected in eight species of birds (Aves, Passeriformes) and in 22 mammalian species: one species of marsupiales (Marsupialia), 3 species of carnivores (Carnivora), seven species of rodents (Rodentia), two species of rabbits and hares (Lagomorpha), in 8 species of even-toed ungulates (Artiodactyla) and one species of odd-toed ungulates (Perissodactyla). The great majority of species comes from the non-arctic area.

Animals↗

PubMed1995

[Lyme disease].

The history of Lyme disease, a contagious condition caused by Borrelia burgdorferi transmitted to man by ticks offers infectiologists a formidable lesson on how medicine progresses. Clinical description started in Europe at the turn of the century with Pick's description of what was then labelled chronic atrophic acrodermatitis. Fifty years later Hauser noted the affection was transmitted by ticks. Independently, Afzelius, then Lipschutz, described erythema chronicum migrans and its relationship with tick bites. Neurological involvement was also described with the skin signs. These early dermatological descriptions suddenly came into the limelight in 1975 when an epidemia of arthritis occurred in children in Lyme, Connecticut, USA. Many of the affected children had erythema chronicum migrans. Based on these observations and an epidemiological analysis of the epidemia, Steele and co-workers defined "Lyme disease" as a rheumatological disorder commonly associated with erythema chronicum migrans and sometimes with multiple organ involvement. In 1982 Borgdorfer suggested that tick bites transmitted a Spirochaeta which was later authentified as the causal agent: Borrelia burgdorferi. Immunofluorescence and ELISA tests were rapidly developed for the diagnosis of infection by this germ which is very difficult to culture. Antibiotic curative treatment was immediately available and in 1991 a consensus conference established recommendations for treatment of isolated and disseminated forms. Antibiotic prophylaxis is not necessary but rapid extraction of the tick after the bite can prevent the disease as transmission from tick to man takes several hours. And medical progress continues. Work is now being conducted on evaluating the extent of late neurological manifestations, on developing polymerase chain reaction methods to identify B. burgdorferi infection in specific organs and on developing a vaccine.

Humans↗

PubMedDate not supplied

[Gastric syphilis. Report on first 2 cases in Cuba].

Two 28 and 25 year old female patients were studied. Endoscopic analysis of their stomachs showed an hyperemic mammillation picture, neoplasia compatible, in the first patient, and an ulcerous lesion in the second one. Both findings were the result of a positive serological test to discard syphilis. All this, together with a cutaneous and epidemiological history, represented an assumed diagnosis of a secondary stage syphilis with gastric disorders. Although it was not necessary, one case was submitted to a specific spirochaeta stain, but results were negative. The true diagnosis resulted from a penicillin treatment, which provoked a cutaneous-visceral history regression as well as a diminishing in V.D.R.L. titles.

Adult↗

PubMed1993

[Gastric ulcers: a rare manifestation of secondary syphilis].

Gastric lesions in the progression of syphilis are very rare and they have several forms of presentation resembling lesions of different nature. We describe a case of secondary syphilis in an heterosexual patient, with persistent gastric ulcers and hepatic affection, in which the evolution and the observations with laparotomy suggested neoplasia. In the gastric biopsy, an infiltration of plasmatic and neutrophil cells was observed and in the hepatic biopsy, necrotic nodules and granulomas. The spirochaeta-specific tinction, although not indispensable, confirms the diagnosis of gastric syphilis. The previous presence of a syphilitic chancre and typical cutaneous lesions are of great help in suspecting a luetic process.

Humans↗