最終更新日:2006年02月01日

細胞培養への新たな汚染微生物?

References

  1. Ciftcioglu,N. and Kajander,E.O. Interaction of nanobacteria with cultured mammalian cells, Pathophysiology, 4: 259-270, 1998.

      Notes: Abstract Nanobacteria were recently isolated from human blood and commercial fetal bovine serum (FBS) and were located in the ce-2 subgroup of proteobacteria based upon their 16S rRNA gene sequence. They can be cultured even in the absence of mammalian cells, and have extraordinary properties, Iike very slow growth rate and an impermeable cell wall, making their detection difficult by standard microbiological techniques. Sin(~ they are present in FBS, and thus in cell cultures, it is essential to clarify their effects on cultured mammalian cells. In this study, we show that four out of six nanobacterial isolates from different sera exerted a cytotoxic effect on 3T6 fibroblasts verilied by M~ 13-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide] viability assay, lactate dehygrogenase (LDH) release and by direct microscopy. The cytotoxic effect of nanobacteria was attenuated after they had been subcultured several times. The cytotoxic effect was similar with all tested murine and human fibroblastoid cell lines. Differential interference contrast and electron microscopy, and FITC staining with specific monoclonal antibodies indicated selective, possibly receptor-mediated adherence, followed by internalization and cytotoxicity in the 3T6 fibroblasts used as a model in these interaction studies. Thus, nanobacteria have a special way of invading mammalian cells: they trigger cells that are not normally phagocytic to engulf them. These organisms seem to be an important cause for cell vacuolization, poor thriving and unexpected cell lysis, problems frequently encountered in mammalian cell culture. C 1998 Elsevier Science B.V.
      Reference ID: 5879

  2. Kajander,E.O. and Ciftcioglu,N. Nanobacteria: an alternative mechanism for pathogenic intra- and extracellular calcification and stone formation, Proc. Natl. Acad. Sci. U. S. A, 95: 8274-8279, 1998.

      Notes: Department of Biochemistry and Biotechnology, University of Kuopio, POB 1627, Fin-70211, Kuopio, Finland OlaviKajander@ukufiFAU - Kajander, E O
      Abstract: Calcium phosphate is deposited in many diseases, but formation mechanisms remain speculative. Nanobacteria are the smallest cell-walled bacteria, only recently discovered in human and cow blood and commercial cell culture serum. In this study, we identified with energy-dispersive x-ray microanalysis and chemical analysis that all growth phases of nanobacteria produce biogenic apatite on their cell envelope. Fourier transform IR spectroscopy revealed the mineral as carbonate apatite. The biomineralization in cell culture media resulted in biofilms and mineral aggregates closely resembling those found in tissue calcification and kidney stones. In nanobacteria-infected fibroblasts, electron microscopy revealed intra- and extracellular acicular crystal deposits, stainable with von Kossa staining and resembling calcospherules found in pathological calcification. Previous models for stone formation have led to an hypothesis that elevated pH due to urease and/or alkaline phosphatase activity is a lithogenic factor. Our results indicate that carbonate apatite can be formed without these factors at pH 7.4, at physiological phosphate and calcium concentrations. Nanobacteria can produce apatite in media mimicking tissue fluids and glomerular filtrate and provide a unique model for in vitro studies on calcification
      Reference ID: 5866

  3. Abbott,A. Battle lines drawn between 'nanobacteria' researchers, Nature, 401: 105, 1999.

      Notes: news, about nanobacteria
      Reference ID: 5860

  4. Abbott,A. Researchers fail to find signs of life in living particles., Nature, 408: 394, 2000.

      Abstract: News, Nanobacterium
      Reference ID: 5859

  5. Cisar,J.O., Xu,D.Q., Thompson,J., Swaim,W., Hu,L., and Kopecko,D.J. An alternative interpretation of nanobacteria-induced biomineralization, Proc. Natl. Acad. Sci. U. S. A, 97: 11511-11515, 2000.

      Notes: Oral Infection and Immunity Branch, and Cellular Imaging Core, National Institute of Dental and Craniofacial Research, National Institutes of Health, Bethesda, MD, 20892, USA johncisar@nihgovFAU - Cisar, J O
      Abstract: The reported isolation of nanobacteria from human kidney stones raises the intriguing possibility that these microorganisms are etiological agents of pathological extraskeletal calcification [Kajander, E. O. & Ciftcioglu, N. (1998) Proc. Natl. Acad. Sci. USA 95, 8274-8279]. Nanobacteria were previously isolated from FBS after prolonged incubation in DMEM. These bacteria initiated biomineralization of the culture medium and were identified in calcified particles and biofilms by nucleic acid stains, 16S rDNA sequencing, electron microscopy, and the demonstration of a transferable biomineralization activity. We have now identified putative nanobacteria, not only from FBS, but also from human saliva and dental plaque after the incubation of 0.45-microm membrane-filtered samples in DMEM. Although biomineralization in our "cultures" was transferable to fresh DMEM, molecular examination of decalcified biofilms failed to detect nucleic acid or protein that would be expected from growth of a living entity. In addition, biomineralization was not inhibited by sodium azide. Furthermore, the 16S rDNA sequences previously ascribed to Nanobacterium sanguineum and Nanobacterium sp. were found to be indistinguishable from those of an environmental microorganism, Phyllobacterium mysinacearum, that has been previously detected as a contaminant in PCR. Thus, these data do not provide plausible support for the existence of a previously undiscovered bacterial genus. Instead, we provide evidence that biomineralization previously attributed to nanobacteria may be initiated by nonliving macromolecules and transferred on "subculture" by self-propagating microcrystalline apatite
      Reference ID: 5867

  6. Pretorius,A.M., Sommer,A.P., Aho,K.M., and Kajander,E.O. HIV and nanobacteria, HIV. Med., 5: 391-393, 2004.

      Notes: National Health Laboratory Services, Department of Medical Microbiology, University of the Free State, Bloemfontein, South Africa gnvrampmd@mailuovsaczaFAU - Pretorius, A-M
      Reference ID: 5887

  7. Harasawa,R., Kurematsu,M., Tanabe,H., and Mizusawa,H. Self-propagating calciferous particles detected in a human cell line Kasumi-6 (JCRB1024), Invitro Cellular & Developmental Biology, Animal., Accepted: 2005.

      Reference ID: 5952

  8. Rawal,B.D. and Pretorius,A.M. "Nanobacterium sanguineum"--is it a new life-form in search of human ailment or commensal: overview of its transmissibility and chemical means of intervention, Med. Hypotheses., 65: 1062-1066, 2005.

      Notes: Human Ailments Research Independent Services (HARIS), San Diego, CA 92129, USA bhupatrawal@yahoocomFAU - Rawal, Bhupat D
      Abstract: Morphological, cultural, and immuno-histochemical characteristics of "Nanobacterium sanguineum" (NB) described in the literature are reviewed. NB is reported to be a motile, Gram negative organism that divides by binary fission within a calcium-coated slimy shell; this yeast-like shell replicates by budding. It measures between 20 and 200 nm with a unique structure containing 16S ribosomal RNA. NB has been observed by electron microscopy in coronary artery plaques (CAD) and in kidney stones (KS) found in renal diseases. On the basis of supportive literature, we suggest that NB is not only present in the human body but also has auxiliary association with human ailments without a specific etiological role; anti-NB antibody has been detected in subjects with calcified lesions and inflammation in diverse ailments including choriodecidual inflammation in pregnancy, ovarian cancers, arthritis and even Alzheimer's disease. More recent report on the detection and vertical transmission of NB antigen and anti-NB antibody in HIV-infected mothers supports the view that NB might be an important opportunistic infective agent contributing to HIV pathology; we note that the presence of viable and transmitting NB was not studied and suggest further studies to establish vertical transmission of NB in HIV-infected persons. On the basis of the foregoing we suggest that NB possibly exacerbates human ailments and raise the question: Is NB a new life-form in search of human ailment or a commensal organism? Recognizing the presence of NB in the human body, we discuss clinical trials, reported in the literature relevant to its eradication, with a rectal suppository containing very high amounts of disodium EDTA and tetracycline. We suggest that tetracycline in this formulation acted in combination with EDTA, more as a chelating agent than an antibiotic; oxytetracycline- a non-chelating form of tetracycline-does not inhibit or kill NB. Evaluation of anti-NB effect of orally administrable and potentially safer as well as therapeutically more acceptable chelating agent -ascorbic acid, acting alone or in combination with antibiotics-that eradicates another slime forming bacterium - Pseudomonas aeruginosa - in children with cystic fibrosis, is suggested
      Reference ID: 5951

  9. Rossa,C., Jr., Marcantonio E Jr, Santos,L.A., Boschi,A.O., and Raddi,M.S. Cytotoxicity of two novel formulations of calcium phosphate cements: a comparative in vitro study, Artif. Organs, 29: 114-121, 2005.

      Notes: Department of Periodontics, School of Dentistry at Araraquara, State University of Sao Paulo, Araraquara, Sao Paulo, Brazil crossa@umicheduFAU - Rossa, Carlos Jr
      Abstract: The purpose was to evaluate the cytotoxicity of two novel formulations (alpha and beta) of calcium phosphate cements. Positive control, represented by a commercial hydroxyapatite cement, and negative control were included for comparative purposes. A continuous lineage of fibroblastic cells was used, and the effect of the tested materials on both cell proliferation and viability was assessed by counting cell number on hemocytometer and by the trypan blue exclusion test, respectively. Study design attempted to simulate clinical use by allowing direct and indirect contact of cells and cements. Results were analyzed by the Kruskal-Wallis test and indicated that the beta formulation was extremely cytotoxic (P < 0.001), because this material induced the greatest reduction on cell proliferation and viability. The alpha formulation behaved similarly to the positive control regarding its effect on cell proliferation and viability. Thus, it is concluded that alpha formulation has promise for further evaluation of its behavior in vivo
      Reference ID: 5861

  10. Wen,Y., Li,Y.G., Yang,Z.L., Wang,X.J., Wei,H., Liu,W., Miao,X.Y., Wang,Q.W., Huang,S.F., Yang,J., Kajander,E.O., and Ciftcioglu,N. Detection of nanobacteria in serum, bile and gallbladder mucosa of patients with cholecystolithiasis, Chin Med. J. (Engl. ), 118: 421-424, 2005.

      Notes: Laboratory of Hepatobiliary Disease, Second Xiangya Hospital, Central-South University, Changsha 410011, China wenyu2861@yahoocomcnFAU - Wen, Yu
      Reference ID: 5886

  11. Wood,H.M. and Shoskes,D.A. The role of nanobacteria in urologic disease, World J. Urol., (online first): 1-4, 2006.

      Notes: Glickman Urological Institute, Cleveland Clinic Foundation Cleveland, Mail Code: A100 OH, 9500 Euclid Avenue, Cleveland, OH 44195, USA, dshoskes@maccom
      Abstract: Recent data proposing an extremely small, self-replicating agent termed "nanobacteria" has raised a great deal of controversy within the scientific community. Since these agents have been isolated within the genitourinary tract, much research has focused attention on the potential role these particles may play in the development of urologic pathology, including polycystic kidney disease, renal calculi, and chronic prostatitis. Recent clinical research targeting these agents has proven effective in treating some patients with refractory category III prostatitis (chronic pelvic pain syndrome). This article reviews the current state of nanobacteria research and explore where these particles may impact urologic disease
      Reference ID: 5950