She froze. It was that sensation.

Asakura felt just a little anxious.

There was still one thing she could not explain. Why had Kiyomi's mitochondria rebelled? Why not Asakura's, or Toshiaki's? Why Kiyomi?

Was it simply a question of polymorphism? Everyone carried slightly different genes.

Had Kiyomi's simply been different in such a way as to allow mitochondrial hyperactivity?

If that were so, what Asakura was about to do did not guarantee that mitochondria would never rise again. As long as people could be born with genetic mutations like Kiyomi's, there was a possibility that mitochondria could evolve in them. If that was so, was it not useless to hinder their evolution?

Asakura had no answers. Maybe so, maybe not.

All she could do with any a.s.surance was to kill these cells.

"The party is winding down now. They're all asking for you," said the young man from beyond the door.

Asakura smiled, then switched on the autoclave.

NOTES.

EcoR I and BamH I.

Used in genetic engineering to cut strands of DNA. EcoR I and BamH I specifically digest (or recognize) the DNA motifs GAATTC and GAGCTC.

clean bench.

A workstation enclosed in gla.s.s into which one places only the hands for sterilized handling of materials. Purified air pa.s.ses into the clean bench via a filtering system.

NIH3T3.

An immortal cell line cultured from the epithelial (i.e. fetal) linings of lab mice.

Normal rodent cells do not divide more than ten times in a cultivation environment.

Cancerous cells, on the other hand, divide indefinitely. NIH3T3 cells are known for their ability to propagate like cancerous cells while exhibiting traits of their healthy counterparts.

Their name pays homage to the American National Inst.i.tutes of Health (NIH), where they were first isolated.

retinoid receptor.

Proteins that bind to a chemical compound (retinoid) similar to Vitamin A. B-oxidation enzymes A series, or chain, of enzymes that draws out energy by decomposing fatty acids. This response can be observed, for example, in respiratory activity, in which B-oxidation enzymes utilize oxygen to consume fatty acids. B-oxidation enzymes exist in mitochondria and even in peroxisomes (one of four organelles in a cell).

hybridoma Cells used to artificially fuse cancer cells and lymphocytes. They are particularly useful for cellular research, as they preserve the nature of lymph cells while multiplying indefinitely.

red indicator solution Putting color into cultivation liquid is standard practice to determine its acidity or alkaline level (pH).Yellow=Acidic, Red=Neutral, and Purple=Alkaline. The condition of the cells is clearly elucidated from its color.

primary culture A process by which cells are extracted from internal organs (human or rodent) and then cultivated. When pre-gathered cells such as NIH3T3 are cultivated, the initial stages are not technically deemed a primary culture.

cancer gene production Whenever toxins such like radiation or carcinogens give rise to genetic mutations, cell division is affected, ultimately causing cancer. There are currently over 100 types of cancerous genes known, with new mutations always being discovered.

immuno-suppressants These drugs suppress the body's natural refusal of foreign matter (in this case, a donated organ). At the same time, however, this makes a recipient more easily p.r.o.ne to bacterial infection, which can sometimes mean the difference between life and death.

HEPES buffer solution A reagent used to prevent changes in a given solution's pH level. HEPES is virtually nontoxic and is ideal for maintaining a chemically neutral environment.

collagenase Any group of proteolytic enzymes that decomposes collagen and gelatin (i.e. cellular proteins), centrifuge A machine by which diluted cells are collected through the use of centrifugal force.

Fifty G's is the usual amount of force applied by the machine, which spins at 700 rpm, creating an artificial gravity 50 times that of the earth's surface. In this context, being gentle means to separate the cells carefully without damaging them.

Eppendorf tubes Plastic receptacles that can hold up to 2 ml. Their resilience makes them ideal for use in centrifuges and are used often in genetic engineering. Costing only pennies each, they can be disposed of freely after use. Though originally a product of its namesake company in Germany, they are now manufactured by various other suppliers.

stirrer A capsule-shaped mixing tool, consisting of a magnet covered in teflon. The stirrer is placed into beaker, into which the solution to be mixed is poured. The beaker is then placed onto a special machine. Another magnet inside the motor spins around, causing the stirrer to spin inside the beaker.

supernatant Clear fluid above sediment or precipitate in a test tube pipettman A device used to extract an exact amount of a given solution anywhere from 0.2 to 1000 micro liters. In genetic engineering, the usual amount used is between 10 and 100 uL.

One uL is equal to one cubic millimeter in volume.

-80 .

Cells can be preserved by freezing them while still alive. This requires a slow cool down in a special preservation liquid, dropping the temperature by one degree per minute. In this way, the cells suffer minimal damage.

Kiyomi's liver cells were glowing Cells treated with collagenase appear as glowing globules when viewed under a phase contrast microscope. Dead or dying cells are opaque and a cell's condition can be judged according to its luminosity. If left for a number of hours, cells aggregate to the bottom of the flask and lose their brightness.

cytosol In addition to the nucleus and mitochondria, cytosol is another important cellular component. Mitochondria are actually stuck to a mesh structure called tubulin, so they are unable to move around freely. In longer cells such as nerve cells, they unite with these "motor proteins" to achieve movement. Free-swimming mitochondria are thus rare.

Kiyomi The name Kiyomi is comprised of two characters, the first meaning 'holy' and the second 'beauty' . When the first character is used in conjunction with that for 'night', the resulting compound means 'Christmas Eve'.

six-well plate.

There are various sizes of plates which have small depressions (wells). The number of available wells ranges from 6 to 96. In a 6-well plate, 2 mL can be put into each 3.S cm diameter well.

MOM19.

Genes coding for a large portion of mitochondrial proteins can be found in the nucleus: the nuclear gene products generated as precursor proteins must be imported into mitochondria. MOM19, a protein in the mitochondrial membrane, is essential for this importation.

cloned.

In a first generation cultivation, the qualities of each cell differ slightly, because the inherent nature of each cell changes during the culturing process. In order to acquire uniform cell groups, a succession of cells is replicated from a single cell. This is called cell cloning.

There are innumerable methods for cell cloning, but the so-called "critical dilution" method is the most widely used of all. In this method, one dilutes a cellular solution and cultivates no more than one cell per plate well. Inside each well, the cells that multiply are virtually identical to the cell from which they propagate. Eve 1 and Eve 2 propagated in separate wells. Therefore, each is a unique clone group. This is different from genetic cloning.

Northern Blot.

A way of detecting and quantifying a specific RNA. RNA can be separated into the following sub- cla.s.sifications: ribosome RNA (rRNA), transfer RNA (tRNA), and messenger RNA (mRNA), among others. The person who devised the DNA detection method was named Southern, so his method came to be known as the Southern blot. RNA is, appropriately enough, "Northern". A Western blot detects proteins. There are "Eastern" and "South Western" methods as well. The amount of beta-oxidation enzyme produced is largely determined by the amount of mRNA coding the protein in question.

RT-PCR.

A method by which mRNA is transcribed to DNA and selectively amplified.

clofibrate A drug that lowers cholesterol in the blood. Clofibrate also acts as a B-oxidation catalyst.

citric acid cycle One of several metabolic pathways in mitochondria, it fuels the reactions that generate sugar, fatty acids, and many amino acids.

cell bank A facility where various types of cell cultures are registered and from which cellular samples are sent out to researchers upon request.

fibroblasts.

Long and thin as their name implies, these cells are relatively easy to culture and can divide about SO times in human biological systems. Even with the best equipment, one can only get liver cells to survive for about 1-2 weeks. It is much easier to cultivate fibroblasts to maintain a high cell count, but here Toshiaki wants to use liver cells as per standard research practice.

needle biopsy.

The most direct method of determining the condition of a transplanted organ. A needle is inserted into the patient's kidney (in Mariko's case) and a small tissue specimen taken. A minute sample of the extracted tissue is stained, then observed under a microscope.

Mitochondrial Eve.

In 1987, Cann and Wilson proposed the theory that we are all descended from a single woman in Africa whom they dubbed Mitochondrial Eve. However, this does not mean that Mitochondrial Eve lived in isolation, but implies only that humanity originated in her and that she thrived with many others.

hot wind.

Mitochondria are the birthplace of energy. They make use of enzymes to combine with ATP. The energy used to consummate this synthesis is stored as electricity and is called an electron transport system (ETS).The potential for one cubic centimeter of mitochondrial activity is 200,000 volts. When we exercise, an emission of heat can also occur directly in mitochondria themselves. This process is what causes fever. The proteins that facilitate this discharge are called "uncoupling proteins" and are found in 307 amino acids. If all mitochondria in the entire body simultaneously discharged in this manner, the heat produced would be unfathomable.

Their membranes fused, first the outer, then the inner.

There is evidence to suggest that when cells fuse together, each mitochondrion actually fuses as well. However, there are also times when the latter does not occur because of mitochondrial abnormalities. This is usually dependent on the conditions in which the fusion takes place.

replicating them or programming them to die at will In the cells of "higher" life forms, death is systematic. Unneeded cells kill themselves gradually over time. This programmed death (also called apoptosis) is hindered by certain proteins that exist chiefly in mitochondria. That this programmed death signal emanates from mitochondria is unmistakable. The manipulation of cells by mitochondria is therefore not entirely unthinkable.

Drosophila.

A fruit fly used extensively in genetic research.

She scowled at the man, then made his heart stop like She did to the last one.

Heart disease has been known to occur from miniscule changes in mitochondrial DNA.

Eve 1 would therefore have to manipulate very little to accelerate such a process.

PCR.

Polymerase Chain Reaction. A technique for amplifying DNA sequences in vitro by separating the DNA into two strands and incubating it with oligonucleotide primers and DNA polymerase. This process can amplify a specific sequence of DNA by as many as one billion times and is important in biotechnology, forensics, medicine, and genetic research.

thermal cycler.

A device used in PCR to replicate specific DNA strands using polymerase. Researchers can investigate DNA extracted from a single human hair or trace of any bodily fluid, and this has forensic uses in criminal cases. The genes of the dinosaurs in Jura.s.sic Park were restored in this way.

polymorphism.

Individual genetic differences.

end.