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 energydispersive 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 and͞or 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 f luids and glomerular filtrate and provide a unique model for in vitro studies on calcification.The formation of discrete and organized inorganic crystalline structures within macromolecular extracellular matrices is a widespread biological phenomenon generally referred to as biomineralization. Mammalian bone and dental enamel are examples of biomineralization involving apatite minerals. The molecular basis of mineralization remains largely unknown (1). Recently, bacteria have been implicated as factors in biogeochemical cycles for mineral formation in aqueous sediments (2, 3). The principal constituent of modern authigenic phosphate minerals in marine sediments is carbonate (hydroxy)fluorapatite Ca 10 (PO 4 ) 6-x (CO 3 ) x (F,OH) 2ϩx . Microorganisms are capable of depositing apatite outside thermodynamic equilibrium in sea water. They can segregate Ca from Mg and actively nucleate carbonate apatite by means of specific oligopeptides under conditions pH Ͻ 8.5 and [Mg]:[Ca] Ͼ 0.1 (4). Such conditions are also present in the human body.We have discovered nanobacteria in human and cow blood that are cytotoxic in vitro (5) and in vivo (6). They have been deposited in DSM (no. 5819-5821; Braunschweig, Germany). Nanobacteria possess unusual properties, making their detection difficult with standard microbiological methods. Although they typically had diameters of 0.2-0.5 m, they passed through 0.1-m filters probably because tiny forms (0.05-0.2 m) were also observed in transmission electron microscopy (TEM) (7). Nanobacteria were poorly disruptable, stainable, fixable, and exceptionally resistant to heat (8, 9). Their doubling time was about 3 days. High doses of ␥-irradiation or aminoglycoside antibiotics prevented their multiplication. According to the 16S rRNA gene sequence (EMBL X98418 and X98419), na...
Selenium (Se) is known to affect the immune system, and decreased Se/levels in blood of patients with moderate or severe psoriasis have been reported. In this study, the effect of Se/supplementation (400 micrograms/day for 6 weeks as Se/yeast, containing about 70% selenomethionine, SeMet) on skin and blood Se/content, on skin glutathione peroxidase activity and on various chemical and immunological parameters of blood and skin was investigated in 7 psoriatic patients. Before the SeMet/supplementation, serum and blood Se/levels were at the normal range, but they increased 42/45% during the Se/dosage, while zinc levels remained unchanged. Se/dependent glutathione peroxidase activity in both normal and lesional psoriatic skin remained unchanged during the trial, although a small net Se/uptake was detected. At the same time, a slight but statistically significant increase in the number of CD4+ T/cells was observed in the reticular dermis of the psoriatic lesions whereas the numbers of CD8+, CD11c+, and CD1+ cells were not significantly altered. Also, a relatively high number of patients (3 out of 7) showed a strongly reduced number of gamma/delta T/lymphocytes or increased CD8+ T/cells (2 patients) in peripheral blood. However, SeMet/supplementation was not related to these abnormalities or to the number of other peripheral blood immunocytes or to serum immunoglobulin levels. In addition, no marked effect on the clinical condition of the patients was observed. This pilot study suggests that SeMet may be able to modulate the immunological mechanism of psoriatic lesions by increasing the number of CD4+ T/cells.
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