History of the

great
invention in
science
"God didn’t make
anything so
complicated that it
can’t be worked out"
Barnet
t
Rosen
berg(19
Mitosis cell division
Magnetic and Electric Field Lines
Idea !

• To study the effect of electric fields on

Cell Division (like Mitosis)
• Started with Escherichia coli bacterial
cells
Experimental Setup

• An audio oscillator was used to supply

sinusoidal voltages of 50 to 100,000
cycles/second (c/s). These signals were
amplified by a conventional audio power
amplifier and then sent to a pair of
platinum electrodes in the chamber
In The First Experiment…
• The electric field (1,000 c/s), After 1
hour, cell growth was slowing down
• After 2 hours, the bacterial population
was almost completely washed out,
The voltage was turned off
• After 8 more hours, the population
density of the bacteria returned to its
previous value
Result ?
When Freq. was 500 - 6,000 c/s

• Bacterial cells stopped dividing and

started to elongate and forming long
filaments
• After the voltage was removed, the cells
continued to increase in length, but after
2 hours had elapsed, cell division started
to occur again.
Effect of Frequency applied

• At 500 c/s, filamentous growth was at a

maximum
• At 6,000 c/s, no filaments could be seen
• For 6,000 to 100,000 c/s no filament
could be seen even after 24 hr
 Role of atmosphere

• Filamentous growth occurred at the

frequencies mentioned above only when
oxygen was present
• If nitrogen or helium, were bubbled
through the cell, no effect was observed
Control Experiments
• The filamentous growth can be caused by
‫ ٭‬Dyes such as methylene blue and penicillin
‫ ٭‬Transfer to an unaccustomed medium
‫ ٭‬Osmotic pressure changes
‫ ٭‬Near ultraviolet irradiation
‫ ٭‬Magnesium deficiency or excess
‫ ٭‬Temperature
‫ ٭‬pH
The application of an electric field to the
bacterial medium might have led to an
electrolysis reaction and that the
chemical products of this reaction might
have affected bacterial cell division
ELECTRODE BACTERIA and
and CULTURE CULTURE
MEDIUM MEDIUM
• The application of the electric current
was not itself responsible for the
observed effects on bacterial growth
• Rather that the electric current led to the
formation of a new chemical species that
affected bacterial elongation
• Oxygen had to be present for bacterial
elongation to occur
Is an Oxidizing agent?
• It gives positive result to the potassium
iodide-starch test
M2+ + 2 I- M0 + I 2 redox reaction
I- + I2 I3 - acid-base reaction

M2+ + 3 I- M0 + I3- net reaction

• Ordinary medium gave no reaction with
potassium iodide and starch
 Intensity of the blue color
corresponded to the frequency of the
applied voltage
The frequency was 500 c/s, the blue
color was most intense
The frequency was 6,000 c/s, the blue
color was not detectable
Who is the mystery oxidizing agent?
• Several possible oxidizing agents could be
created from the medium during electrolysis
hypochlorite ion (ClO-)
chlorite ion (ClO2-)
chlorate ion (ClO3-)
perchlorate ion (ClO4-)
hydrogen peroxide (H2O2)
hydroxylamine (NH2OH)
persulfate ion (S2O82-)
Negative results-(Starch iodide Test)
• Phosphate ion (PO43-)
• Sulfate ion (SO42-)
• Phosphate ion + glucose
• Phosphate ion + sulfate ion + glucose
• Sodium sulfate (Na2SO4)
• Sodium carbonate (Na2CO3)
Positive results-(Starch iodide Test)

• Ammonium sulfate ((NH4)2SO4)

• Ammonium carbonate ((NH4)2CO3)
• Ammonium chloride (NH4Cl)
• Sodium chloride (NaCl) gave a faint
positive response
Clue!

Platinum electrodes could be attacked

by an acidified chloride solution
during electrolysis, generating PtCl62-
Later Results…
• (NH4)2PtCl6 was shown positive
potassium iodide-starch test(>100 ppm)
• A later account suggests that (NH4)2PtCl6
at first caused cell death then later, after
standing on a laboratory shelf for a few
weeks, produced a small number of
filaments
 Cobalt (Co)
Iridium (Ir)
Nickel (Ni)
Osmium (Os)
Palladium (Pd)
Rhodium (Rh)
Ruthenium (Ru)
Platinum-containing compounds
(NH4)2PtIVCl6+ uv light PtIV(NH3)2Cl4
Action of Cisplatin

Cisplatin coordinates to DNA and that

this coordination complex not only
inhibits replication and transcription of
DNA, but also leads to programmed cell
death (called apoptosis)
Cisplatin in cell level
Binding site in Base Pairs…
Geometrical Isomer
• In vitro studies on both prokaryotic and
eukaryotic cells revealed that DNA adducts of
both cisplatin and trans-DDP blocked the
action of DNA polymerase
• In vivo studies showed that cisplatin and
trans-DDP inhibited replication equally well
• DNA replication is not the only factor
important for the clinical activity of cisplatin
The cytotoxic activity of cisplatin may
arise from the cell’s inability to repair
DNA damage caused by cisplatin.
• The cell detects DNA damage by the action
of damage recognition proteins
• HMG-domain proteins bind cisplatin–DNA
adducts in vitro
• In vivo assays on yeast shown that HMG-
domain proteins are important for the
activity of cisplatin:
• These effects may also be in operation in
mammalian cells
Role of HMG domain proteins
1. HMG domain containing transcription
factors bind preferentially to the
cisplatin–DNA adducts, they could
wreak havoc with the transcriptional
machinery
2. When HMG domain proteins bind to
the cisplatin–DNA adducts, the adducts
would not be recognized by the repair
machinery
DNA-Cisplatin-HMG adduct
2G Drugs
3G drugs
Drug Resistance
• Acquired resistance
– Drug is initially beneficial but becomes
ineffective over time
• Intrinsic resistance
– Drug is ineffective from the outset
Resistance Mechanism

• Decreased Intracellular Accumulation

• Sulfur-Containing Macromolecules
• Increased DNA Repair
Side Effects

• Thrombocytopenia
• Leukopenia
• Anemia
• Nephrotoxicity
• Ototoxicity
 Cisplatin was approved as an anticancer
drug in 1978 by the Food and Drug
Administration
$250 million in royalty !!!
Synthetic path way of Cisplatin
"In the fields of observation,
chance favors only the mind
that is prepared " - Louis
Pasteur
THANK YOU