DISCUSSION

The importance of chromosome aberrations for evolution and their association with
human health have been recognized for almost a century. However, the mechanisms
involved in the formation of chromosome aberrations are still not well understood.
Ionizing radiation is most efficient in inducing chromosome aberrations and the basic
principles of aberration formation were laid out in the early 1930’s, even though the
molecular structure of the eukaryotic chromosome was not known at that time.
Techniques to prepare and stain chromosomes have improved gradually in the last
decades, parallel with increased ability to identify and quantify chromosome aberrations,
thus leading to a better understanding of their origin. Use of colchicine, squash technique
for plant chromosomes, hypotonic shock for mammalian chromosomes, various banding
techniques in the 1960’s, sister chromatid differentiation in the early 1970’s and
fluorescence in situ hybridization (FISH) in the late 1980’s have been main break-
throughs enabling us to study chromosome aberrations in great detail. Especially the
increasing availability of whole chromosome-specific, armspecific, region-specific,
centromere- and telomere-specific DNA probes has revolutionized the ways to look at
chromosomes and their aberrations.1

DNA damage, due to environmental factors and normal metabolic processes inside the
cell, occurs at a rate of 1,000 to 1,000,000 molecular lesions per cell per day.2 While this
constitutes only 0.000165% of the human genome's approximately 6 billion bases (3
billion base pairs), unrepaired lesions in critical genes (such as tumor suppressor genes)
can impede a cell's ability to carry out its function and appreciably increase the likelihood
of tumor formation..

DNA damage can be subdivided into two main types,endogenous damage such as attack
by reactive oxygen species produced from normal metabolic byproducts (spontaneous
mutation), especially the process of oxidative deamination; exogenous damage caused by
external agents such as ultraviolet [UV 200-300nm] radiation from the sun other radiation
frequencies, including x-rays and gamma rays ,hydrolysis or thermal disruption certain
plant toxins,human-made mutagenic chemicals, especially aromatic compounds that act
as DNA intercalating agents ,cancer chemotherapy and radiotherapy.The vast majority of
DNA damage affects the primary structure of the double helix; that is, the bases
themselves are chemically modified. These modifications can in turn disrupt the
molecules' regular helical structure by introducing non-native chemical bonds or bulky
adducts that do not fit in the standard double helix. Unlike proteins and RNA, DNA
usually lacks tertiary structure and therefore damage or disturbance does not occur at that
level. DNA is, however, supercoiled and wound around "packaging" proteins called
histones (in eukaryotes), and both superstructures are vulnerable to the effects of DNA
damage.

In the present study the positive control, cyclophosphamide has shown significant
changes in the chromosomal structure compare to control.The different aberration
observed were gaps,breaks ring,ploidy,stikiness,exchanges and fragment.Therefore ,it is
proved as a mutagenic agent and justifies it as positive control in our study.This drug has
been used as a mutagen in no of mutagenicity tests.3,4

Cyclophosphamide is an inactive cyclic phosphamide ester of mechlorethamine. It is

transformed via hepatic and intracellular enzymes to active alkylating metabolites, 4-
hydroxycyclophophosphamide, aldophosphamide, acrolein and phosphoramide mustard.
Cyclophosphamide causes prevention of cell division primarily by cross-linking DNA
strands. It is considered to be cell cycle phase-nonspecific. 5

**Oxidative stress is produced in cells by oxygen-derived species resulting from cellular

metabolism and from interaction with cells of exogenous sources such as carcinogenic
compounds, redox-cycling drugs and ionizing radiations. DNA damage caused by
oxygen-derived species including free radicals is the most frequent type encountered by
aerobic cells. DNA damage caused by oxygen-derived species including free radicals is
the most frequent type encountered by aerobic cells. When this type of damage occurs to
DNA, it is called oxidative DNA damage and it can produce a multiplicity of
modifications in DNA including base and sugar lesions, strand breaks, DNA-protein
cross-links and base-free sites. Accurate measurement of these modifications is essential
for understanding of mechanisms of oxidative DNA damage and its biological effects.
Numerous DNA lesions have been identified in cells and tissues at steady-state levels and
upon exposure to free radical-generating systems. Data accumulated over many years
clearly show that oxidative DNA damage plays an important role in a number of disease
processes. Thus, oxidative DNA damage is implicated in carcinogenesis and
neurodegenerative diseases such as Alzheimer’s disease. There is also strong evidence for
the role of this type of DNA damage in the aging process. The accumulation of oxidative
DNA damage in non-dividing cells is thought to contribute to age-associated diseases.
DNA damage is countered in cells by DNA repair, which is a basic and universal process
to protect the genetic integrity of organisms. The genomes of organisms encode DNA
repair enzymes that continuously monitor chromosomes to correct DNA damage.
Multiple processes such as base- and nucleotide-excision pathways exist to repair the
wide range of DNA damages. If left unrepaired, oxidative DNA damage can lead to
detrimental biological consequences in organisms, including cell death, mutations and
transformation of cells to malignant cells. Therefore, DNA repair is regarded as one of
the essential events in all life forms. There is an increasing awareness of the importance
of oxidative DNA damage and its repair to human health. Thus, it becomes exceedingly
important to understand, at the fundamental level, the mechanisms of oxidative DNA
damage, and its processing by DNA repair enzymes as well as how unrepaired DNA
lesions may lead to cytotoxicity, mutagenesis and eventually to diseases and aging. **