RESUMEN
Leeway space preservation in the mixed dentition is a well-documented method of space management. In the mandibular arch it may be saved for utilisation in the correction of minor anterior crowding by the placement of a passive lower lingual arch (LLA) during the transition from the mixed dentition to the permanent dentition.
Asunto(s)
Dentición Mixta , Maloclusión/terapia , Diseño de Aparato Ortodóncico , Aparatos Ortodóncicos , Mantenimiento del Espacio en Ortodoncia/instrumentación , Diente Canino/patología , Arco Dental/patología , Humanos , Incisivo/patología , Mandíbula/patología , Odontometría/métodos , Soportes Ortodóncicos , Alambres para Ortodoncia , Ortodoncia Interceptiva/instrumentación , Erupción Dental/fisiología , Exfoliación Dental/fisiopatología , Diente Primario/fisiologíaRESUMEN
OBJECTIVE: Chlamydia trachomatis and Chlamydophila (Chlamydia) pneumoniae are known triggers of reactive arthritis (ReA) and exist in a persistent metabolically active infection state in the synovium, suggesting that they may be susceptible to antimicrobial agents. The goal of this study was to investigate whether a 6-month course of combination antibiotics is an effective treatment for patients with chronic Chlamydia-induced ReA. METHODS: This study was a 9-month, prospective, double-blind, triple-placebo trial assessing a 6-month course of combination antibiotics as a treatment for Chlamydia-induced ReA. Eligible patients had to be positive for C trachomatis or C pneumoniae by polymerase chain reaction (PCR). Groups received 1) doxycycline and rifampin plus placebo instead of azithromycin; 2) azithromycin and rifampin plus placebo instead of doxycycline; or 3) placebos instead of azithromycin, doxycycline, and rifampin. The primary end point was the number of patients who improved by 20% or more in at least 4 of 6 variables without worsening in any 1 variable in both combination antibiotic groups combined and in the placebo group at month 6 compared with baseline. RESULTS: The primary end point was achieved in 17 of 27 patients (63%) receiving combination antibiotics and in 3 of 15 patients (20%) receiving placebo. Secondary efficacy end points showed similar results. Six of 27 patients (22%) randomized to combination antibiotics believed that their disease went into complete remission during the trial, whereas no patient in the placebo arm achieved remission. Significantly more patients in the active treatment group became negative for C trachomatis or C pneumoniae by PCR at month 6. Adverse events were mild, with no significant differences between the groups. CONCLUSION: These data suggest that a 6-month course of combination antibiotics is an effective treatment for chronic Chlamydia-induced ReA.
Asunto(s)
Artritis Reactiva/tratamiento farmacológico , Azitromicina/administración & dosificación , Infecciones por Chlamydia/complicaciones , Chlamydia trachomatis/aislamiento & purificación , Doxiciclina/administración & dosificación , Rifampin/administración & dosificación , Adulto , Anciano , Antibacterianos/administración & dosificación , Antibacterianos/efectos adversos , Antibióticos Antituberculosos/administración & dosificación , Antibióticos Antituberculosos/efectos adversos , Artritis Reactiva/microbiología , Artritis Reactiva/patología , Azitromicina/efectos adversos , Chlamydia trachomatis/genética , Chlamydophila pneumoniae/genética , Chlamydophila pneumoniae/aislamiento & purificación , Enfermedad Crónica , ADN Bacteriano/genética , Método Doble Ciego , Doxiciclina/efectos adversos , Quimioterapia Combinada , Femenino , Humanos , Articulaciones/patología , Masculino , Persona de Mediana Edad , Placebos , Prohibitinas , Estudios Prospectivos , Rifampin/efectos adversos , Resultado del TratamientoRESUMEN
Mandibular canine impaction and transmigration have serious consequences for the patient, as removal of the tooth or teeth in question is often the only solution. The loss of one or both mandibular canines complicates orthodontic treatment. Early warning signs of mandibular canine ectopia are explored in this paper as well as how to assess the potential for displacement, impaction and/or transmigration. This paper highlights the value of interceptive treatment once the early signs of an aberrant mandibular canine have been detected.
Asunto(s)
Diente Canino/patología , Mandíbula/patología , Erupción Ectópica de Dientes/diagnóstico , Diente Premolar/fisiología , Diagnóstico Precoz , Humanos , Maloclusión/diagnóstico , Maloclusión/prevención & control , Maloclusión/terapia , Ortodoncia Interceptiva , Factores de Tiempo , Erupción Dental/fisiología , Erupción Ectópica de Dientes/prevención & control , Erupción Ectópica de Dientes/terapia , Exfoliación Dental/fisiopatología , Diente Primario/fisiología , Diente Impactado/diagnóstico , Diente Impactado/prevención & controlRESUMEN
This paper focuses on the identification of ectopic eruption patterns of the maxillary canines from the dental ages of approximately 8 to 12 years. The timing and suitability of interceptive treatment in pre-adolescents are discussed.
Asunto(s)
Diente Canino/fisiopatología , Erupción Ectópica de Dientes/diagnóstico , Erupción Ectópica de Dientes/cirugía , Extracción Dental , Diente Primario/cirugía , Niño , Odontología General , Humanos , Maloclusión Clase II de Angle/complicaciones , Maxilar , Radiografía Panorámica , Resorción Radicular/etiología , Erupción Ectópica de Dientes/complicaciones , Movilidad Dentaria/etiología , Diente Impactado/etiologíaRESUMEN
The mixed dentition pantomogram is routinely used in paediatric patients. This paper discusses the value of the pantomogram for early identification of problems in dental development during the mixed dentition stage. Aspects regarding dental maturity, leeway space, the sequence of eruption of the permanent teeth, anomalies and the development of the canines will be reviewed.
Asunto(s)
Dentición Mixta , Odontogénesis/fisiología , Radiografía Panorámica , Determinación de la Edad por los Dientes , Arco Dental/diagnóstico por imagen , Humanos , Maloclusión/prevención & control , Erupción Dental/fisiologíaRESUMEN
Rarely, tumour necrosis factor (TNF)alpha antagonist therapy has been associated with de novo psoriasiform eruptions. This is unusual in that these same drugs are used to treat psoriasis. Most of these cases involve the palms and soles, yet palmoplantar pustular psoriasis represents only 1.7% of all cases of psoriasis. Keratoderma blenorrhagicum is a psoriasiform rash that occurs primarily on the palms and soles of some patients with reactive arthritis. It is grossly and histologically indistinguishable from pustular psoriasis. Chlamydia trachomatis is a common aetiological agent for reactive arthritis, and in vitro studies have shown that chlamydial replication is inversely proportional to TNFalpha levels. Three patients taking TNFalpha antagonists are presented who developed such lesions and who were found to be positive for C trachomatis DNA in the affected skin. It is proposed that these psoriasiform lesions may not be psoriasis, but rather keratoderma blenorrhagicum.
Asunto(s)
Antirreumáticos/efectos adversos , Artritis Reactiva/tratamiento farmacológico , Infecciones por Chlamydia/complicaciones , Queratosis/inducido químicamente , Factor de Necrosis Tumoral alfa/antagonistas & inhibidores , Adalimumab , Adulto , Anticuerpos Monoclonales/efectos adversos , Anticuerpos Monoclonales Humanizados , Artritis Reactiva/patología , Artritis Reumatoide/tratamiento farmacológico , Artritis Reumatoide/patología , Chlamydia trachomatis , Etanercept , Femenino , Humanos , Inmunoglobulina G/efectos adversos , Infliximab , Receptores del Factor de Necrosis Tumoral , Piel/patologíaRESUMEN
Infraocclusion of primary molar teeth is a relatively common clinical finding and decision-making concerning the successful management of such cases can present a challenge to the general dental practitioner. This paper reviews the epidemiology, aetiology, diagnosis, treatment rationale and includes flow charts for easy reference to the various treatment options for infraoccluded primary molar teeth.
Asunto(s)
Maloclusión/terapia , Diente Molar/patología , Diente Primario/patología , Terapia Combinada , Árboles de Decisión , Humanos , Maloclusión/diagnóstico , Ortodoncia Interceptiva , Planificación de Atención al Paciente , Diente no Erupcionado/diagnóstico , Diente no Erupcionado/terapiaRESUMEN
An investigation was conducted to determine the outcome of nonextraction edgewise orthodontic treatment of thirty-five caucasian patients with Class II molar relationships (13 division 1 and 22 division 2 cases). Twenty-three females and 12 males aged between 10 and 16 years old were treated using Class II intermaxillary elastics worn with 0.016" x 0.022" stainless steel archwires and without extra-oral traction. Thirty cephalometric landmarks were identified and digitized, and used to calculate 41 parameters. Space analyses were conducted on pretreatment study models. Most of the cases had between 0-4 mm space shortage in both maxillary and mandibular dentitions. Results of this study indicated that the mean SNA angle decreased by 1.580 degrees, and that this change was largely due to posterior movement of point A. The mean pre-treatment ANB value was 4.490 degrees which decreased to a mean posttreatment value of 2.810 degrees (p<0.05). No significant changes in the growth direction of the jaws were found, showing that the use of Class II intermaxillary elastics in this sample had no detrimental effect on this parameter (p<0.05). The SNB angle increased by a mean of 0.110 degrees, which can be attributed to anterior mandibular growth. The dentoalveolar changes included a mean overjet reduction of 3.816mm (mean posttreatment overjet was 2.2mm) and proclination of the lower incisors relative to the APo line and the mandibular plane, probably caused by the use of Class II intermaxillary elastics. The lower lip advanced relatively more than the upper lip, and this contributed to an improvement of the lip relationship. The amount of nose growth observed during the study was normal for this age group, and compared favourably with other data in the literature.
Asunto(s)
Maloclusión Clase II de Angle/terapia , Ortodoncia Correctiva/métodos , Adolescente , Cefalometría/métodos , Niño , Femenino , Humanos , Masculino , Ortodoncia Correctiva/instrumentación , Estudios Retrospectivos , Resultado del TratamientoRESUMEN
Two column tests were performed in conditions emulating vertical flow beneath the leachate table in a biologically active landfill to determine dominant transport mechanisms occurring in landfills. An improved understanding of contaminant transport process in wastes is required for developing better predictions about potential length of the long term aftercare of landfills, currently measured in timescales of centuries. Three tracers (lithium, bromide and deuterium) were used. Lithium did not behave conservatively. Given that lithium has been used extensively for tracing in landfill wastes, the tracer itself and the findings of previous tests which assume that it has behaved conservatively may need revisiting. The smaller column test could not be fitted with continuum models, probably because the volume of waste was below a representative elemental volume. Modelling compared advection-dispersion (AD), dual porosity (DP) and hybrid AD-DP models. Of these models, the DP model was found to be the most suitable. Although there is good evidence to suggest that diffusion is an important transport mechanism, the breakthrough curves of the different tracers did not differ from each other as would be predicted based on the free-water diffusion coefficients. This suggested that solute diffusion in wastes requires further study.
Asunto(s)
Monitoreo del Ambiente/métodos , Contaminantes del Suelo/análisis , Residuos Sólidos/análisis , Instalaciones de Eliminación de Residuos , Difusión , Agua Subterránea/análisis , Porosidad , Eliminación de ResiduosRESUMEN
In the yeast Saccharomyces cerevisiae, a phenotypically identical stringent response is induced by either nutritional downshift or starvation for a required auxotrophic amino acid (aa); in each case, the response selectively includes transcriptional curtailment for the mitochondrial (mt) genome. We have shown previously that the downshift-induced mt stringent response is governed by changing cellular cyclic AMP (cAMP) levels, via a mt cAMP-dependent protein kinase. In contrast, we demonstrate here that cAMP levels are not altered in yeast following starvation for a required aa, and we use in vitro mt transcription assays with organelles from wild-type and mutant strains to confirm that the aa starvation-induced mt stringent response is not governed by cAMP. Rather, such stringent organellar transcriptional attenuation may result from altered availability of an unidentified small molecule which is probably a product of the cytoplasmic and/or mt protein synthesis systems.
Asunto(s)
Aminoácidos/metabolismo , Regulación Fúngica de la Expresión Génica/fisiología , Mitocondrias/metabolismo , Saccharomyces cerevisiae/metabolismo , Transcripción Genética , Medios de Cultivo , AMP Cíclico/metabolismo , Proteínas Quinasas Dependientes de AMP Cíclico/metabolismo , Regulación hacia Abajo/fisiología , Mutación , Saccharomyces cerevisiae/crecimiento & desarrolloRESUMEN
Culture of Chlamydia trachomatis from synovial tissues/fluids from Reiter's syndrome (RS) patients frequently yields negative results. However, we have identified chlamydial RNA at that site in such patients, suggesting that viable organisms may be present. Here we define the cellular location of chlamydia within the synovium via in situ hybridization. Using a chlamydial ribosomal RNA-directed probe, we show that synovial tissue from culture-negative RS patients gives strong hybridization which is often localized to a subsynovial cell layer, rather than to the synovial lining; in some cases, hybridizing cells are dispersed through the synovium. All hybridization signal is located within host cells, indicating that infectious extracellular elementary bodies are rare or absent. These data confirm the extensive intracellular presence of inapparent chlamydia in the synovia of RS patients and provide some insight into the usual culture negativity of synovial tissues for the organism.
Asunto(s)
Artritis Reactiva/microbiología , Chlamydia trachomatis/aislamiento & purificación , Membrana Sinovial/microbiología , Adulto , Artritis Reactiva/patología , Infecciones por Chlamydia/diagnóstico , Femenino , Humanos , Hibridación in Situ , Masculino , Persona de Mediana Edad , Membrana Sinovial/citología , Membrana Sinovial/patologíaRESUMEN
A very abundant and dG + dC rich DNA satellite from primate Cebus capucinus has been characterized in its cytogenetic and biochemical properties with the purpose of studying the correlation between the staining properties of heterochromatin and the base composition of the corresponding very repetitive DNA. The staining techniques, as well as incorporation of base analogues, show that the heterochromatin segments of C. capucinus chromosomes correspond to a dG + dC-rich satellite. This satellite was detected and isolated by centrifugation in density gradient, radioactively labelled and localized by in situ hybridization on heterochromatin segments.
Asunto(s)
Cebidae/genética , Cebus/genética , ADN Satélite/análisis , Desoxicitidina/análisis , Desoxiguanosina/análisis , Desoxirribonucleasas de Localización Especificada Tipo II , Animales , Bandeo Cromosómico , Enzimas de Restricción del ADN/metabolismo , Hibridación de Ácido NucleicoRESUMEN
PURPOSE: MRL/MpJ-+/+ (MRL/+) and MRL/MpJ-lpr/lpr (MRL/lpr) mice show spontaneous development of a T-cell-driven lacrimal gland inflammation that is a model for Sjögren syndrome. The lacrimal gland lesions in these mice were evaluated by quantitative RT-PCR for selected cytokine mRNA for the relative contributions of T-helper (Th)1 versus Th2 immune responses and by RT-PCR and immunohistochemistry for the contribution of the interleukin (IL)-2/IL-2 receptor (IL-2R) autocrine pathway. METHODS: RNA was isolated from lacrimal glands of MRL/+ mice ages 1 to 9 months and from MRL/lpr mice ages 1 through 5 months, and competitive RT-PCR was used to quantify mRNA for the cytokines IL-2, -4, -10, and -12 and interferon (IFN)-gamma. Frozen sections of lacrimal glands from MRL/+ and MRL/lpr mice ages 2 through 5 months were stained for the IL-2R. RESULTS: IL-2 and -12 mRNA transcripts were below the limit of detection (<10(-3) fg/pg hypoxanthine phosphoribosyl transferase gene; HPRT) in both MRL/+ and MRL/lpr mice of all ages. When detectable, IFN-gamma transcripts were present in low amounts and were below the limit of detection in most samples. IL-4 transcripts were present in 100- to 1000-fold greater amounts than IFN-gamma transcripts. IL-10 transcripts were detectable in both MRL/+ and MRL/lpr mice. IL-2R typically was detected on less than 10% of lymphocytes infiltrating lacrimal gland lesions in both substrains. CONCLUSIONS: On the basis of RT-PCR for cytokine mRNA, autoimmune lacrimal gland lesions in MRL/+ and MRL/lpr mice appear to be largely Th2-mediated. There does not appear to be a direct role for the IL-2/IL-2R autocrine pathway within the microenvironment of the lacrimal gland.
Asunto(s)
Citocinas/fisiología , Aparato Lagrimal/inmunología , Síndrome de Sjögren/inmunología , Células Th2/inmunología , Animales , Técnicas para Inmunoenzimas , Ratones , Ratones Endogámicos MRL lpr , ARN Mensajero/metabolismo , Receptores de Interleucina-2/fisiología , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Células TH1/inmunologíaRESUMEN
Present or prior infection with any of several common bacterial pathogens has been strongly implicated in the development of Reiter's syndrome (RS). However, because Chlamydia trachomatis is the most common sexually transmitted bacterial pathogen in the western hemisphere, this organism has emerged as a major causative agent of the disease. An important but as yet poorly understood aspect of chlamydia-related RS is the recurrence of active arthritis in the absence of any overt infection or reinfection with the organism. In this article, a large body of published clinical and experimental observations strongly suggesting that some chlamydial infections in humans can be inapparent--not detectable by standard culture or antibody-based laboratory screening methods--is reviewed. The authors' own initial molecular genetic studies, which support that contention, are summarized. Based on these data, the argument is developed that not only does inapparent chlamydial infection occur in humans, but such infections may have significant consequences in terms of the pathogenesis of RS, including the possible causation of recurrent episodes of disease in the absence of active infection. The therapeutic implications of potential inapparent chlamydial infections as they relate to RS are discussed.
Asunto(s)
Artritis Infecciosa/etiología , Artritis Reactiva/microbiología , Infecciones por Chlamydia , Chlamydia trachomatis , Membrana Sinovial/microbiología , HumanosRESUMEN
The etiology of RS is not clear, but there is a strong correlation with infectious episodes and some genetic factor(s). Chlamydia have emerged as the most common pathogen associated with RS. We have presented evidence that chlamydia or its antigens may be present in the synovium and may be important in the pathogenesis of RS. The possibility of latent chlamydial infection has also been discussed. Although previous attempts to treat RS with antibiotics were not encouraging, recent reports suggest some favorable effects from antibiotic therapy. There is still a need to further assess the state of the infectious agent and to consider new, more effective regimens.
Asunto(s)
Artritis Reactiva , Artritis Reactiva/complicaciones , Infecciones por Chlamydia , Antibacterianos/uso terapéutico , Antígenos Bacterianos/análisis , Artritis Reactiva/etiología , Artritis Reactiva/microbiología , Chlamydia/inmunología , Chlamydia/aislamiento & purificación , Infecciones por Chlamydia/tratamiento farmacológico , Humanos , Membrana Sinovial/inmunología , Membrana Sinovial/microbiologíaRESUMEN
An infectious etiology has been suggested for Reiter's syndrome (RS) because the disease has often been observed to follow episodes of urethritis or dysentery. Despite demonstrations of bacterial antigens in the synovial tissues of RS patients, it is not clear whether viable organisms are present in the synovium in any particular stage of this disease. Furthermore, it is not clear how either viable organisms or their product(s) might reach the joints. Infection with the bacterium Chlamydia trachomatis is the most common sexually transmitted disease in the United States, and as such this organism has emerged as a primary pathogen associated with RS. Previous work from our group has shown that synovial biopsy tissues from a majority of RS patients studied show significant levels of apparently intact chlamydial RNA, even when synovial or urethral cultures from the same patients are unequivocally negative for the organism. We show here that inapparent urethral infection with chlamydia occurs with high prevalence in men, and that inapparent cervical infection with the organism occurs at high prevalence in women. These data provide an important link in the relationship between initial chlamydial infection and possible subsequent genesis of RS, and they may give useful insight into mechanisms by which chlamydial infection can lead to development of this disease. Our data argue further that inapparent infection may be a significant factor in pathogenesis for all chlamydia-related diseases, and they suggest that, contrary to current ideas, C. trachomatis can generate disseminated infection.
Asunto(s)
Artritis Reactiva/etiología , Infecciones por Chlamydia/complicaciones , Chlamydia trachomatis , Enfermedades Uretrales/complicaciones , Enfermedades del Cuello del Útero/complicaciones , Femenino , Humanos , Masculino , Hibridación de Ácido Nucleico , Enfermedades Uretrales/microbiología , Enfermedades del Cuello del Útero/microbiologíaRESUMEN
OBJECTIVE: To use standard molecular methods to define the prevalence and metabolic characteristics of Chlamydia trachomatis during infection of fallopian tubes in women with ectopic pregnancies. DESIGN: Polymerase chain reaction (PCR)- and reverse transcription-PCR (RT-PCR)-based assessment of presence of chlamydial DNA and various RNA species in fallopian tube biopsy samples. SETTING: Hospital and molecular genetics laboratory. PATIENTS: Ten women of varying ages, each presenting with ectopic pregnancy. MAIN OUTCOME MEASURE(S): Positive signal in specific chlamydia-directed PCR and RT-PCR assays. RESULT(S): Nucleic acid preparations from 7 of the 10 fallopian tube patient samples were PCR-positive for C. trachomatis DNA. Each of the 7 PCR-positive samples also showed the presence of several transcripts from the bacterium, including primary transcripts from the ribosomal RNA operons. CONCLUSION(S): A higher proportion of ectopic pregnancies than was believed previously may be attributable to infection of the fallopian tubes by C. trachomatis. The presence of various chlamydial RNA molecules suggests that viable, metabolically active bacteria were present in fallopian tubes of the patients studied.
Asunto(s)
Chlamydia trachomatis/crecimiento & desarrollo , Trompas Uterinas/microbiología , Embarazo Ectópico/microbiología , Adulto , Femenino , Células HeLa , Humanos , Reacción en Cadena de la Polimerasa/métodos , Embarazo , Reacción en Cadena de la Polimerasa de Transcriptasa InversaRESUMEN
OBJECTIVE: Giant cell arteritis (GCA) is a vasculitis predominantly affecting medium- and large-sized arteries. Recent data show the co-localization of dendritic cells and Chlamydia pneumoniae in vascular biopsies from GCA patients. Here we define the topographical relation of dendritic cells and these activated T-cells to determine the antigen presenting cell in GCA, and to examine several auxiliary biochemical and genetic aspects relating to the role of bacteria such as C. pneumoniae in eliciting GCA. METHODS: 18 paraffin-embedded temporal artery biopsy specimens from 14 patients with GCA that were PCR-positive for C. pneumoniae were examined by two-color immunohistochemistry for the topographical relationship between dendritic cells and activated T-cells. In addition the presence of GTP-binding proteins. Tumor necrosis factor alpha (TNF alpha), and Toll-like receptor 4 (TLR4) was investigated. 15 temporal artery specimens from 10 patients without GCA served as controls. RESULTS: In all GCA specimens, dendritic cells co-localized in the immediate vicinity of activated CD4+ Talin-expressing T cells, and these were predominantly found in granulomatous infiltrates. Confocal microscopy confirmed the cell-cell contact of dendritic cells with activated T cells. Results further showed that RhoA and Rac1 were predominantly present in the region of granulomatous infiltrates. TNF alpha production and expression was found in dendritic cells and macrophages, predominantly in granulomatous infiltrates and in endothelial cells of the vasa vasorum dispersed in the adventitial and medial layers of the temporal artery. No control specimens showed TNF alpha expression. More than 95% of dendritic cells were positive for TLR4; macrophages and endothelial cells localized in the adventitia showed TLR4 production. CONCLUSIONS: The immediate co-localization of dendritic cells and activated T cells indicate a high probability that the former represent the antigen presenting cells in GCA. In addition, because of the presence of Rho A and Rac1 in the granulomatous infiltrates, we speculate that they provide the right environment for cell-cell contact and adhesion, and that they may promote the internalization of bacteria. TNF alpha is expressed at high levels in the granulomatous infiltrates of temporal artery specimens from patients with GCA. Since TLR4 is produced in the same cell types, and predominantly in the adventitial layer of the temporal artery, we suggest that these receptors are coupled to signal transduction pathways that control TNF alpha expression.
Asunto(s)
Linfocitos T CD4-Positivos/inmunología , Adhesión Celular/inmunología , Células Dendríticas/inmunología , Arteritis de Células Gigantes/inmunología , Arteritis de Células Gigantes/patología , Arterias Temporales/patología , Anciano , Presentación de Antígeno , Biopsia , Chlamydophila pneumoniae/inmunología , Femenino , Arteritis de Células Gigantes/microbiología , Humanos , Masculino , Glicoproteínas de Membrana/inmunología , Receptores de Superficie Celular/inmunología , Arterias Temporales/microbiología , Receptor Toll-Like 4 , Receptores Toll-Like , Factor de Necrosis Tumoral alfa/inmunología , Proteína de Unión al GTP rac1/inmunología , Proteína de Unión al GTP rhoA/inmunologíaRESUMEN
Both clinical and epidemiological data suggest that inapparent infection by Chlamydia trachomatis occurs in humans. To confirm and study such infections, we developed a hybridization screening system directed toward chlamydial ribosomal RNA (rRNA). Six restriction endonuclease fragments derived from the cloned rrnA operon of chlamydial serovar L2(434) were tested as hybridization screening probes, but only one fragment encoding the 5' portion of the 16s rRNA gene plus some upstream flanking sequence was both sensitive and highly specific in such experiments. In Northern slot blot assays, hybridization analyses with this fragment as probe routinely detected one picogram or less of chlamydial RNA when that RNA was bound to membranes alone or as part of a mixture with a vast excess of mammalian RNA. The probe did not hybridize to RNA from mammalian and relevant bacterial sources but did hybridize to rRNA from B (ocular) and E (genital) serovars of C. trachomatis. Experiments using RNA from conjunctival biopsies and standard conjunctival swab samples from cynomolgus monkeys showed that the probe reliably distinguishes between known chlamydia-infected and uninfected samples. This suggests that it may be useful for clinical screening. Characterization assays for the RNA-directed probe screening system in this monkey model of trachoma provide initial molecular evidence that ocular chlamydial infection may persist longer than previously thought, based solely on direct fluorescence antibody assay (DFA) and culture analyses.
Asunto(s)
Infecciones por Chlamydia/diagnóstico , Chlamydia trachomatis/genética , Sondas de ADN , ARN Ribosómico/análisis , Animales , Northern Blotting , Humanos , ARN Ribosómico/genética , Ratas , Mapeo Restrictivo , Homología de Secuencia de Ácido Nucleico , Especificidad de la EspecieRESUMEN
The authors have shown that protein antigens, RNA, and DNA from Chlamydia trachomatis are present in synovial tissues of patients with Reiter's syndrome (RS). However, those studies gave no insight into the host cell type involved or the precise tissue location of the bacteria. To address such issues, the authors developed an in situ hybridization system to detect chlamydia, and they used that system to examine synovial biopsies from a patient with RS and a patient without RS. The in situ system uses a previously described digoxigenin-labeled DNA probe that hybridizes with chlamydial 16S rRNA sequences in paraformaldehyde-fixed samples. Control studies with chlamydia-infected and uninfected HeLa cells confirmed that the in situ system is as sensitive as is direct fluorescence cytology for detection of the organism. Morphology of host and chlamydia cells is preserved after hybridization. Studies using synovial tissue from an osteoarthritis patient produced no in situ hybridization signal, but similar hybridization to tissue from a culture-/direct fluorescence cytology- negative RS patient had a strong intracellular signal for chlamydia within a subsynovial cell layer. These in situ hybridization results confirm the extensive presence of chlamydia in synovia and extend the authors' earlier observation that chlamydia RNA is present in the synovia of patients with RS. The data also confirm their electron microscopy studies, indicating that chlamydia are intracellular in synovial tissue, and they further show that infected host cells are located beneath the synovial lining.