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            Nanobacterium sanguineum is a small, slowly growing bacterium that can be cultured from human blood, kidney stones, and calcific vascular wall plaque.  Typical bacteria have a diameter of 1/1000th of a meter and divide rapidly into daughter cells.  Nanobacteria are 10,000 to 100,000 times smaller than typical bacteria, in the 20-200 nanometer (nm) range (1 nanometer is 1/1,000,000,000th of a meter).  Nanobacteria divide very, very slowly, splitting into daughter cells once every three days, 1/10,000th the growth rate of conventional bacteria.  Nanobacteria do not grow in standard bacteriologic culture conditions.  Their small size, slow grow rate, and unusual culture requirements explain why Nanobacterium sanguineum has until now eluded detection, as they can only be seen under an electron microscope.

An isolated Nanobacterium in cell culture (Figure 1) demonstrates a round to D-shaped configuration, termed coccoid by microbiologists.  Initially 20 nm is size, it is seen to “grow” in mammalian culture media, related to the elaboration of a goey biofilm (Figure 2).  Over time, the biofilm hardens to cover the Nanobacterium like an “igloo”.  Figure (3) is a cross-section through a mature, isolated Nanobacterium; the spicules surrounding the cell body are composed of carbonate appetite, the principle form in which abnormal, extra-skeletal calcium is found in humans.  Isolated Nanobacteria tend to coalesce, merging their biofilms to form a common shelter, which protects the Nanobacteria from predators such as heat, radiation, the immune system, and most antibiotics.  Figure (4) demonstrates Nanobacteria elaborating biofilm which is beginning to thicken and calcify.  Figure (5) is a cross-section through a nanobacterial colony, surrounded by its calcific shell, and figures (6 & 7) are scanning electron microscope “outside” views, demonstrating a raspberry shaped “shelter” that houses and protects the Nanobacteria.

Nanobacteria can be cultured from the blood of medical students (5%), blood donors (15%), dialysis patients (80%), and untreated cardiovascular patients.  In one study, urine cultures for Nanobacterium sanguineum were positive in 30% of healthy men.  Kajander and Ciftcioglu, the Finnish researchers and Nobel Prize nominees who first described Nanobacterium sanguineum, obtained kidney stones from 30 consecutive patients, ground them up, and then examined the stones under the electron microscope.  What they found (Figure 8) looks a lot like figures (6 & 7), colonies of Nanobacteria within their calcific shelters.  Hydrochloric acid was then applied to demineralize the kidney stone samples, and from this pulverized, acid treated kidney stone powder they could culture Nanobacterium sanguineum.  If you inject Nanobacteria into an animal, it will concentrate in the kidney cells, from there migrate into the “tubules” (ducts into which urine is formed from the filtered blood), and then appear in the urine.  Kidney stones don’t form spontaneously in the urine, but as precipitates of calcium and other minerals upon dysfunctional, calcified kidney tubule cells (Figure 9), much like stalactites hanging down from the roof of a cave.  Nanobacterium sanguineum initiates this calcification.  Eventually this mass of calcified Nanobacteria shelters and precipitated minerals will break off from its attachment to the kidney and migrate into the ureter (the tube connecting the

kidney to the bladder) where it can lodge, producing the pain of “renal colic”.  5% of women and 10% of men will experience a kidney stone at some point in their life; Nanobacteria sanguineum appears to be the culprit.  Nanobacterium sanguineum can be cultured from dental pulp stones and calcified pineal glands (the brain center that controls our sleep-wake cycle).  It is becoming clear that abnormal calcification in our body is related to Nanobacterium sanguineum. 

Laslo Puskas, a Hungarian researcher, applied calcium specific stains and fluorescent, Nanobacteria-specific monoclonal antibodies (which attach only  to Nanobacteria), to microscopic sections of carotid and aortic atherosclerotic     plaque.  66% of the specimens “lite up” for Nanobacteria (that is, the Nanobacteria specific antibodies attached to a Nanobacterium or a Nanobacterial particle).  100% of the                                                                                                            specimens stained for calcium, in a  “calcospherule” pattern.  The diameter of the calcospherules was similar to that of the calcific Nanobacteria shelters that can be grown in cell culture.  Applying EDTA, a calcium-removing agent, enhanced the antibody response (by removing the calcific shell, you allow the antibodies greater exposure to the Nanobacteria).  Puskas was then able to culture Nanobacterium sanguineum colonies from 63% of the specimens.  Thus live Nanobacteria sanguineum colonies are present in human atherosclerotic plaque, and correspond to areas    of vascular wall calcification.

Our arteries are calcium-free at birth.  While calcium deposition within the vascular wall is not unusual with aging, it is not normal.  We do not want calcium within our blood vessel walls!  The Ultrafast CT Scanner can localize and quantitate calcium within our coronary arteries.  As a general rule, the more calcium present in our arteries, the more likely we are to have obstructive coronary artery disease, placing us at risk for heart attack or stroke.  In relation to age, gender, and risk factor status, your CT calcium “score” can be used to predict the likelihood that you have a blocked coronary artery.  Of interest, in coronary patients, the calcium score progresses at a rate of 45% per year, roughly the same rate at which kidney stone calcification progresses. 

Kajander and Ciftcioglu found that Nanobacterium sanguineum is resistant to heat, hydrochloric acid, high-dose gamma irradiation (150 megarads), and most antibiotics, while it can be inhibited or killed by Tetracycline (an antibiotic with metal chelating properties) and EDTA (a calcium and heavy metal chelating agent).  With the knowledge that Nanobacterium sanguineum is associated with vascular wall calcification, and that the degree of calcification correlates with the severity of coronary artery disease, nanobacterial researcher Dr. Gary Mezo reasoned that anti-Nanobacterial therapy might be of value in the treatment of coronary artery disease, as well as other disease states associated with abnormal, extra-skeletal calcification.  His NanobacTX-ACEs study protocol involves the nightly administration of NanobacTX, (a rectal EDTA suppository, coupled with an oral prescription powder and Tetracycline taken by mouth).  The idea is that the EDTA dissolves the calcific shell that is protecting the Nanobacterial colony, allowing the tetracycline to then kill the Nanobacteria.  The powder component of the prescription is designed to optimize the efficacy of the EDTA and to reduce soft “uncalcified” plaque.  This treatment “sterilized” the blood, such that while on therapy, antibodies to Nanobacteria are present, but Nanobacteria cannot be cultured from the patient’s blood.  Of the 90 patients to complete three months of NanobacTX treatment in the NanobacTX-ACEs single center pilot study, the mean Ultrafast CT calcium score decreased by 59%; in 19 patients calcium could no longer be detected.  It is anticipated that continued treatment will resolve vascular wall calcium in the remaining patients. 

                                                                                                                                                                7/20/01 – James C. Roberts MD FACC