Nanobacteria Research Articles

A potential cause for kidney stone formation during space flights: Enhanced growth of nanobacteria in microgravity

Although some information is available regarding the cellular/molecular changes in immune system exposed to microgravity, little is known about the reasons of the increase in the kidney stone formation in astronauts during and/or after long duration missions at zero gravity (0 g). In our earlier studies, we have assessed a unique agent, nanobacteria (NB), in kidney stones and hypothesized that NB have an active role in calcium phosphate-carbonate deposition in kidney. In this research we studied effect of microgravity on multiplication and calcification of NB in vitro. We examined NB cultures in High Aspect Rotating Vessels (HARVs) designed at the NASA's Johnson Space Center, which are designed to stimulate some aspects of microgravity. Multiplication rate and calcium phosphate composition of those NB were compared with NB cultured on stationary and shaker flasks. Collected aliquots of the cultures from different incubation periods were analyzed using spectrophotometer, SEM, TEM, EDX, and x-ray diffraction techniques. The results showed that NB multiplied 4.6x faster in HARVs compared to stationary cultures, and 3.2x faster than shaker flask conditions. X-ray diffraction and EDX analysis showed that the degree of apatite crystal formation and the properties of the apatite depend on the specific culture conditions used. We now report an increased multiplication rate of NB in microgravity-simulated conditions. Thus, NB infection may have a potential role in kidney stone formation in crew members during space flights. For further proof to this hypothesis, screening of the NB antigen and antibody level in flight crew before and after flight would be necessary.

HIV and nanobacteria

HIV and nanobacteria

Superadhesion: Attachment of Nanobacteria to Tissues − Model Simulation

Adhesion is common in the bacterial world and is partially mediated by slime. Nanobacteria (NB) seem to attach by slime as well as by the protecting nanocrystalline apatite shell, securing their survival in hostile milieus. Identification of NB in the human heart has brought the particles into the focus of material scientists. In view of the size distribution of the apatite nanovesicles (80−300 nm), simulation of their interaction with tissues can be performed via nanosuspensions and mirror-polished titanium substrates. Due to its wide biocompatibility, titanium is an ideal body tissue substitute. Here we show in a model simulation that the apatite nanovesicles are likely to perform also an anchoring function, specific to the mineral apatite. Anchoring may prevent solitary NB from elimination from the body, and enforce existing toxic potentials. The capacity of the apatite nanoparticles to bind to tissues in aqueous liquids, and the pronounced tendency of NB to form mineralized biofilms, indicate that NB could affect intracardiac fluid forces. Adhesion and protection represent a unique combination, which may stimulate the design of novel drug release systems based on apatite nanovesicles.

Calcification in coronary artery disease can be reversed by EDTA–tetracycline long-term chemotherapy

Atherosclerosis is a complex process with multiple mechanisms and factors contributing to its initiation and progression. Detection and quantification of coronary artery calcium (CAC) scores with electron beam tomography has been shown to correlate with obstructive and nonobstructive coronary artery disease (CAD). Pathogen-triggered calcification could play a role in CAD. Recent reports suggest that infectious blood nanobacteria (NB) emerge to be such a trigger. So far, minimal or no reversal of atherosclerosis has been claimed by therapies with iv ethylenediaminetetraacetic acid disodium salt (EDTA), antibiotics, or other regimens, and therapies for atherosclerosis remain non-curative. We have now combined EDTA with antibiotic tetracycline (comET), an in vitro proven nanobacteriocidal treatment, and tested comET therapy in patients with documented CAD. Three hypotheses were probed: (1) Are NB present in patients with CAD?; (2) Does treatment with comET affect blood NB antigen and serology?; (3) Does a comET decrease CAC scores? One hundred patients with stable CAD and positive CAC scores were enrolled into a 4 month study of comET therapy. ComET therapy is composed of (1) Nutraceutical Powder (Vitamin C, Vitamin B6, Niacin, Folic Acid, Selenium, EDTA, l-Arginine, l-Lysine, l-Ornithine, Bromelain, Trypsin, CoQ10, Grapeseed Extract, Hawthorn Berry, Papain) 5 cm 3 taken orally every evening; (2) Tetracycline HCl 500 mg taken orally every evening; (3) EDTA 1500 mg taken in a rectal suppository base every evening. CAC scoring was repeated at 4 months and serum samples were analyzed for NB antigen and serology at baseline, 2 and 4 months. Complete blood count, metabolic panel, liver function, C-reactive protein (hs-CRP) and lipids were analyzed at baseline and 4 months. Seventy-seven patients completed the study and all patients were positive for NB serology, antigen or both. Responders ( n = 44; 57%) had significant decreases in total CAC scores ( P = 0.001), the average decrease being 14%. Non-responders ( n = 33; 44%) had no change or had increases in CAC scores. Angina was decreased or ablated in 16 of 19 patients (84%). Lipid profiles improved to non-atherogenic direction significantly ( P = 0.001), a remarkable finding in a patient group where 86% were on continuous statin medication already before the trial. No adverse physiologic effects were seen in renal, hepatic, or hematopoetic systems. In conclusion, CAC scores decreased during ComET therapy trial in most CAD patients inferring regression of calcified coronary artery plaque volume. The patients tolerated the therapy well and their angina and lipid profiles improved. Further treatment trials for long term therapy with matched controls are warranted.

Biomineralization Induced by Stressed Nanobacteria

Animal experiments provided direct evidence that myocyte apoptosis may be a causal mechanism of heart failure, suggesting that inhibition of this cell death process may constitute the basis for novel therapies. The data suggested that even inhibition of a small fraction of cardiac myocyte apoptosis could be instrumental in preventing cardiomyopathy. Here we analyze the mechanisms by which nanobacteria (NB) are expected to contribute to the inhibition of cellular functions in the heart. NB are protected by a nanocrystalline apatite shell. Under environmental stress, they produce a slime providing ideal conditions for individual mineralization and rapid formation of giant thrombogenic assemblies. We establish a model based upon a possible synergistic impact of physical and chemical stimuli on NB, exposing the principles of a novel preventive strategy promising to inhibit formation of NB clusters in the circulation.

A preliminary investigation into light-modulated replication of nanobacteria and heart disease.

The purpose of this preliminary study is to evaluate the effect of various wavelengths of light on nanobacteria (NB). NB and mitochondria use light for biological processes. NB have been described as multifunctional primordial nanovesicles with the potential to utilize solar energy for replication. NB produce slime, a process common to living bacteria. Slime release is an evolutionary important stress-dependent phenomenon increasing the survival chance of individual bacteria in a colony. In the cardiovascular system, stress-induced bacterial colony formation may lead to a deposition of plaque. Cultured NB were irradiated with NASA-LEDs at different wavelengths of light: 670, 728 and 880 nm. Light intensities were about 500k Wm(-2), and energy density was 1 x 10(4) J m(-2). Monochromatic light clearly affected replication of NB. Maximum replication was achieved at 670 nm. The results indicate that suitable wavelengths of light could be instrumental in elevating the vitality level of NB, preventing the production of NB-mediated slime, and simultaneously increasing the vitality level of mitochondria. The finding could stimulate the design of cooperative therapy concepts that could reduce death caused by myocardial infarcts.

Inhibition of nanobacteria by antimicrobial drugs as measured by a modified microdilution method.

Compounds from 16 classes of antimicrobial drugs were tested for their abilities to inhibit the in vitro multiplication of nanobacteria (NB), a newly discovered infectious agent found in human kidney stones and kidney cyst fluids from patients with polycystic kidney disease (PKD). Because NB form surface calcifications at physiologic levels of calcium and phosphate, they have been hypothesized to mediate the formation of tissue calcifications. We describe a modified microdilution inhibitory test that accommodates the unique growth conditions and long multiplication times of NB. This modified microdilution method included inoculation of 96-well plates and determination of inhibition by periodic measurement of the absorbance for 14 days in cell culture medium under cell culture conditions. Bactericidal or bacteriostatic drug effects were distinguished by subsequent subculture in drug-free media and monitoring for increasing absorbance. NB isolated from fetal bovine serum (FBS) were inhibited by tetracycline HCl, nitrofurantoin, trimethoprim, trimethoprim-sulfamethoxazole, and ampicillin at levels achievable in serum and urine; all drugs except ampicillin were cidal. Tetracycline also inhibited multiplication of isolates of NB from human kidney stones and kidney cyst fluids from patients with PKD. The other antibiotics tested against FBS-derived NB either had no effect or exhibited an inhibitory concentration above clinically achievable levels; the aminoglycosides and vancomycin were bacteriostatic. Antibiotic-induced morphological changes to NB were observed by electron microscopy. Bisphosphonates, aminocaproic acid, potassium citrate-citric acid solutions, and 5-fluorouracil also inhibited the multiplication of NB in a cidal manner. Insights into the nature of NB, the action(s) of these drugs, and the role of NB in calcifying diseases may be gained by exploiting this in vitro inhibition test system.

Newly Discovered Tiny Bacteria May Cause Kidney Stones.

Last year a new type of bacteria, nanobacteria, was discovered in both human and cow blood. The smallest bacteria with a cell wall yet known, they are