THE DISCOVERY OF NANOBACTERIUM SANGUINEUM
Galileo, Semmelweis, Fleming, Kajander, Ciftcioglu, & Serendipity
Utilizing mathematics and the power of observation, Galileo determined
the order of the planets, but of course he was wrong; the experts of the day all
knew that Earth was the centering body. Semmelweis
had the idea that doctors should pause when walking between the autopsy and
labor and delivery suites, to wash their hands.
Fleming feared an upset stomach when he observed bluish mold on his
sandwich bread, but he had the inquisitive nature to determine its cause.
Kajander and Ciftcioglu were having a bad day, which they turned around
in their favor, and ours.
Olavi Kajander MD PhD and Neva Ciftcioglu PhD were carrying out mammalian cell research at the University of Kuopio in Finland; well they were trying to one day in 1988. A routine mammalian cell culture experiment, using commercially available fetal bovine serum as the growth media, just wasn’t getting off the ground. The cells weren’t thriving and dividing like they should; the cells were sickly and died off before any study could be done. Strange vacuoles were forming up in many of the cells, and these cells subsequently died. Kajander and Ciftcioglu, like all basic cell researchers, had encountered this problem before; sometimes their cell cultures worked, and sometimes they didn’t. All researchers use commercially available, defined mammalian cell lines. They all grow these cells in cell culture media, which contains sterile pooled fetal bovine sera, assumed sterile because it is passed through a 100-micron filter.
In the past, they would discard the sickly cells and start again with a new batch, hoping for better luck. This time, serendipity set it. This time, Kajander and Ciftcioglu set out to determine just why this batch of cells was dying. They and other researchers had observed that sometimes their cell cultures thrived, and sometimes they didn’t. Rapidly dividing mammalian cells always seemed to do well, while slowly dividing cultures were more likely to “fail to thrive”. The problem wasn’t the cells themselves; if it was, then all the cultures would be sick. The problem appeared to be the growth media, fetal bovine serum (FBS), which is pooled from multiple animals. They knew that only 10% of commercially available fetal bovine serum batches supported cell cloning research, where the cells divide quite slowly, so they reasoned that the culprit was a slowly growing infective agent, present in some but not all FBS batches –so they set up an experiment to figure this out, kind of like what Fleming did with his moldy bread.
Pooled fetal bovine serum is sterilized by passage through a 100-micron filter, which should normally remove all fungi and cell walled bacteria. Kajander and Ciftcioglu passed different batches of FBS through a 1-micron filter, and treated it with antibiotics and anti-fungal agents, but cells cultured in the resulting serum still failed to thrive. They knew that Mycoplasma, cell wall deficient organisms, and viruses could pass through a 0.1-micron filter and this antiseptic gauntlet, so they irradiated the FBS with 1 megarad, enough to kill any infective organism – still the cells got sick. Whatever the culprit, it was resistant to all known anti-microbial treatments, 1 megarad of irradiation, and it was really small, smaller than any previously described bacterial agent (to eliminate Nanobacteria, a 0.02 micron or 20 nm filter is required).
The next step was to culture out the culprit – if it was infectious, then it had to demonstrate metabolic function, and it had to grow. They inoculated bacteriologic culture media with different lots of FBS obtained from different supply houses – but nothing grew. Next they inoculated cell culture media, not cells but cell culture media, with aliquots of FBS, under varying conditions of pH, pO2, mineral concentration, etc., and nothing grew. Well, nothing grew at first, but after several weeks, turbidity developed in one of the flasks. This represented the first isolation of Nanobacterium sanguineum. These profound discoveries made them Nobel Prize Nominees in 1998. Fourteen years later we are utilizing Kajander and Ciftcioglu’s discovery. For the last 4 years, Ciftcioglu& Kajander have been working with Dr.Gary Mezo at NanobacLabs in Tampa, Florida. Mezo has developed the world’s first nanobiotic (NanobacTX), to treat Nanobacterium sanguineum infections, and reverse atherosclerosis in our patients. For their scientific knowledge, power of observation, and strength of character needed to ignore the criticism of the experts and persist in the pursuit of science, they clearly deserve the Nobel Prize – and I bet they’ll get it.
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