Characterization of Biofilm Producer Nanobacteria Isolated from Kidney Stones of Some Egyptian Patients.

<b>Background and Objective:</b> Nanobacteria (NB) appear to contribute to many calcifying diseases including kidney stones which represent a common problem with inadequate prevention exist. NB framing itself with a mineral coat that assists as a primary defence shield against the immune system, antibiotics. This study aims to collect and detect nanobes from different kidney stones from patients with active urolithiasis then investigated the anti-nano-bacterial activity of some antibiotics alone or in combination with extracts of irradiated herbs of certain medicinal plants which will represent a new approach to therapy for patients with kidney stones. <b>Materials and Methods:</b> Total of 32 nanobes were isolated from 54 kidney stones. Fourier Transforms Infrared Spectroscopy (FTIR) revealed that calcium and phosphate are the main components of stones. Scanning Electron Microscopy (SEM) with Energy-dispersive X-ray spectroscopy (EDX) and Transmission Electron Microscope (TEM), showed that nanobes were Gram-ve cocci with size ranged from (375:600 nm). The biofilm production ability of nanobes was estimated qualitatively and quantitatively. <b>Results:</b> The results revealed that all were strong biofilm producers. Further, the antibiotic susceptibility test indicates their resistance towards most of the tested antibiotics. Molecular identification of the strong biofilm producer isolates by ribosomal ribonucleic acid (rRNA) revealed that it is indicated by 85.37% to <i>Bartonella apis</i> strain PEB0122. <b>Conclusion:</b> The findings of the current study evidenced that combination treatment between Doxycycline (DO) and water extract of khella exhibited a significant reduction in biofilm formation ability of the strongest producers nanobes. Therefore, this treatment can play a role in enhancing public health, especially with patients who suffer from recurrent kidney stone formation.

Different Approaches to Detect "Nanobacteria" in Patients with Kidney Stones: an Infectious Cause or a Subset of Life?

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.

Isolation, cultivation and identification of nanobacteria from placental calcification.

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.

The role of calcifying nanoparticles in biology and medicine.

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.