RESUMEN
PURPOSE: This research focused on the detection of nanobacteria in kidney stones of 30 Iranian patients without adding fetal bovine serum (FBS) to the culture media. MATERIALS AND METHODS: Nanobacteria were isolated from a nephro-ureterolithiasis extract of the urinary tract and kidney of patients and were cultured in the laboratory. The growth of nanobacteria was monitored using a spectrophotometer, and with inverted microscopy technique, their crystallization was analyzed after two days. The images from atomic force microscopy (AFM), transmission electron microscopy (TEM) and scanning electron microscopy (SEM) indicated the morphology and demonstrated the size of the cultured nanobacteria which is between 60 and 160 nm. Fourier transform infrared spectroscopy (FTIR) and X-ray diffraction (XRD) were used to study the chemical composition, surface functional groups and crystal structure of the igloo-like nanobacteria shell. FTIR spectra in theregion of 1000 to 1200 cm-1 and the XRD peaks provided evidence that the main components of the nanobacteria shell were apatite-based compounds. RESULTS: Nanobacteria infected all the 27 patients with apatite kidney stone, and none of the three patients who had uric acid kidney stone were infected as confirmed by the cultivation of the stones samples. The results showed that nanobacteria might play a fundamental role in the formation of apatite-based kidney stones. CONCLUSION: The biomineralization ability of nanobacteria may lead to calcification of the soft tissues, which in turn may result in other diseases. It is also suggested that nanobacteria may be a factor in calcification-related diseases and disorders with poorly characterized etiologies. This research with its different approaches, clarified significant doubts that nanobacteria act as contaminant, warranting continued investigation of its role in other diseases.
Asunto(s)
Nanopartículas Calcificantes/análisis , Cálculos Renales/química , Adolescente , Adulto , Nanopartículas Calcificantes/aislamiento & purificación , Calcinosis/complicaciones , Cristalización , Femenino , Humanos , Masculino , Microscopía de Fuerza Atómica , Microscopía Electrónica de Rastreo , Microscopía Electrónica de Transmisión , Persona de Mediana Edad , Espectroscopía Infrarroja por Transformada de Fourier , Difracción de Rayos X , Adulto JovenRESUMEN
OBJECTIVE: To compare the results of polymerase chain reaction (PCR) and immunologic methods for the detection of nanobacteria (NB) in the expressed prostatic secretions (EPSs) of patients with type-III prostatitis. METHODS: In total, 150 patients with type-III prostatitis for whom conventional clinical treatment had failed were selected from September 2009 to April 2010. The EPS of each patient was divided into 3 parts, which were used for PCR analysis, indirect immunofluorescence staining (IIFS), and culture and subsequent indirect immunofluorescence staining (CIIFS). RESULTS: PCR analysis has a higher sensitivity than IIFS for the detection of NB in EPSs. Of 83 CIIFS-positive EPS samples, 79 (95.2%) were positive by PCR. Of 67 EPS samples that were negative by CIIFS, 60 (89.6%) were negative by PCR. The sensitivity of PCR for the detection of NB compared with the CIIFS method was 95.2%, with a specificity of 89.6%. The positive predictive value was 91.9%, and the negative predictive value was 93.8%. A comparative evaluation showed no statistically significant difference between PCR and CIIFS in the detection of NB in EPSs. A strong agreement in the positive and the negative results obtained by PCR and CIIFS for NB detection was found for all EPS samples. CONCLUSION: PCR analysis has a higher sensitivity than IIFS for NB detection in type-III prostatitis. PCR can detect nanobacterial infection in type-III prostatitis equally well as CIIFS and offers significant advantages for the rapid, simple, and economical detection of nanobacterial infection in type-III prostatitis.
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Nanopartículas Calcificantes/aislamiento & purificación , Técnica del Anticuerpo Fluorescente Indirecta/métodos , Reacción en Cadena de la Polimerasa/métodos , Próstata/metabolismo , Prostatitis/diagnóstico , Prostatitis/microbiología , Humanos , Masculino , Sensibilidad y Especificidad , Factores de TiempoRESUMEN
BACKGROUND: The purpose of this study was to examine the possible contribution of calcifying nanoparticles to the pathogenesis of placental calcification. METHODS: Calcified placental tissues and distal tissue samples were collected from 36 confirmed placental calcification cases. In addition, 20 normal placental tissue samples were obtained as a control group. All the tissue samples were cultured using special nanobacterial culture methods. The cultured calcifying nanoparticles were examined by transmission electron microscopy (TEM), and their growth was monitored by optical density (OD) at a wavelength of 650 nm. 16S rRNA gene expression of the cultured calcifying nanoparticles was also isolated and sequenced. RESULTS: Novel calcifying nanoparticles wrapped with electron-dense shells between 50 nm to 500 nm in diameter were observed in the extracellular matrix of calcified placental tissues. They were detected in placental villi and hydroxyapatite crystals, and contained "nucleic acid-like materials". After isolation and four weeks of culture, 28 of 36 calcified placental tissue samples showed white granular precipitates attached to the bottom of the culture tubes. OD(650) measurements indicated that the precipitates from the calcified placental tissues were able to grow in culture, whereas no such precipitates from the control tissues were observed. The 16S rRNA genes were isolated from the cultured calcifying nanoparticles and calcified placental tissues, and their gene sequencing results implied that calcifying nanoparticles were novel nanobacteria (GenBank JF823648). CONCLUSION: Our results suggest that these novel calcifying nanoparticles may play a role in placental calcification.
Asunto(s)
Nanopartículas Calcificantes/aislamiento & purificación , Calcinosis/patología , Enfermedades Placentarias/patología , Placenta/química , Adulto , Nanopartículas Calcificantes/genética , Nanopartículas Calcificantes/metabolismo , Calcinosis/genética , Calcinosis/metabolismo , Calcinosis/microbiología , Técnicas de Cultivo , ADN/química , ADN/aislamiento & purificación , Durapatita/química , Femenino , Humanos , Datos de Secuencia Molecular , Tamaño de la Partícula , Placenta/patología , Enfermedades Placentarias/genética , Enfermedades Placentarias/metabolismo , Enfermedades Placentarias/microbiología , Embarazo , ARN Ribosómico 16S/genéticaRESUMEN
AIMS: Placental calcification is a common pathologic condition in obstetrics. To detect the bacteria infection mechanisms for calcification, an experiment was performed to isolate, culture, and identify the nanobacteria in placental calcification. METHOD: Sixteen cases of placental calcification of pregnant women were collected for the purpose of the isolation of nanobacteria, cultivation, and identification of 16S rDNA sequence. RESULT: Under transmission electron microscope, novel oval-shape nanobacteria-like particles (NLP) in extracellular matrix of calcified placenta tissues were found with 50-500 nm in diameter, and among hydroxyapatite crystals aggregation existed. After about 4 weeks of culturing and isolating NLP from these calcified tissues, all calcified placental tissue samples and one adjacent tissue of calcified placental tissue samples showed white granular depositions, which were firmly attached to the bottom of the culture tubes and visible to the naked eyes. In the control group they could not be seen. After PCR was amplified a 1407-bp fragment was obtained and submitted to GenBank after sequencing with accession number JN029830. The 16S rDNA sequence homology between the isolation strain and strain nanobacteria (X98418) was 92% in GenBank. CONCLUSION: For the first time isolated, cultured, and identified nanobacteria in placental calcification indicated that nanobacteria infection is related to placental calcification.
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Nanopartículas Calcificantes/aislamiento & purificación , Calcinosis/patología , Enfermedades Placentarias/patología , Placenta/patología , Adulto , Nanopartículas Calcificantes/genética , Nanopartículas Calcificantes/fisiología , Calcinosis/metabolismo , ADN Bacteriano/análisis , ADN Ribosómico , Matriz Extracelular/ultraestructura , Femenino , Humanos , Placenta/metabolismo , Enfermedades Placentarias/metabolismo , Embarazo , Análisis de Secuencia de ADN , Adulto JovenRESUMEN
OBJECTIVE: The etiology of placental calcification (PC) is lack of research. To detect the bacterial infection mechanisms for PC, the experiment of isolating, culturing and identifying the nanobacteria in PC was done. METHOD: The calcified placental tissues from 18 confirmed PC cases with normal placental tissue samples from 18 cases were studied by transmission electron microscopy (TEM), special nanobacterial culture methods, and identification of 16S rRNA sequence. RESULT: Under transmission electron microscope (TEM), Nanobacteria-like particles (NLP) in extra-cellular matrix (ECM) of calcified placental tissues were found, they were 50-500 nm in diameter, existed aggregation, among hydroxyapatite (HAP) crystals. Isolation and culture of NLP from the calcified tissues with methods described for nanobacteria were successful. All calcified placental tissue samples showed white granular deposition, which were firmly attached to the bottom of the culture tubes visible to the naked eyes. In the control group they could not be seen. According to 16S rRNA gene sequences analysis and was amplified adopting PCR and obtained 1407 bp fragment. Submit to GenBank after sequencing with accession number JN029830. CONCLUSION: Indicating that nanobacteria infection is related with placental calcification.
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Nanopartículas Calcificantes/análisis , Nanopartículas Calcificantes/aislamiento & purificación , Calcinosis/microbiología , Enfermedades Placentarias/microbiología , Adulto , Nanopartículas Calcificantes/genética , Calcinosis/patología , Técnicas de Cultivo de Célula , Células Cultivadas , Femenino , Humanos , Técnicas Microbiológicas , Microscopía Electrónica de Transmisión , Filogenia , Placenta/microbiología , Placenta/patología , Placenta/ultraestructura , Enfermedades Placentarias/patología , Embarazo , ARN Ribosómico 16S/análisis , Adulto JovenRESUMEN
Calcifying nanoparticles (CNPs) (nanobacteria, nanobacteria-like particles, nanobes) were discovered over 25 years ago; nevertheless, their nature is still obscure. To date, nobody has been successful in credibly determining whether they are the smallest self-replicating life form on Earth, or whether they represent mineralo-protein complexes without any relation to living organisms. Proponents of both theories have a number of arguments in favor of the validity of their hypotheses. However, after epistemological analysis carried out in this review, all arguments used by proponents of the theory about the physicochemical model of CNP formation may be refuted on the basis of the performed investigations, and therefore published data suggest a biological nature of CNPs. The only obstacle to establish CNPs as living organisms is the absence of a fairly accurately sequenced genome at the present time. Moreover, it is clear that CNPs play an important role in etiopathogenesis of many diseases, and this association is independent from their nature. Consequently, emergence of CNPs in an organism is a pathological, not a physiological, process. The classification and new directions of further investigations devoted to the role of CNPs in biology and medicine are proposed.