**7. References**

476 Selected Topics in DNA Repair

NHEJ, *Afl* III that digests the coding region of *Renilla* luciferase gene is used and the luciferase can only be expressed after exact repair (Fig. 4A). The linearized reporter DNA fragments are purified, transfected into host cells and examined for luciferase activity as described above. Below is an example of evaluating the effect of areca nut extracts on precise

Fig. 4. HCR assay for non-homologous end-joining (NHEJ) repair.

nucleotides in this region during the end-joining process.

overall (right panel) NHEJ repair.

**5. Conclusion** 

(A) The *Afl* III-digested pRL-CMV is used as a substrate for analyzing precise NHEJ repair activity because of the need of exact joining of the *Renilla* luciferase coding sequence. For overall NHEJ, *Hin*d III that cuts the flanking sequence between CMV promoter and the *Renilla* luciferase gene is used. The expression of luciferase is not affected by loss of a few

(B) The effect of areca nut extracts (ANE, 800 mg/ml for 24 h) on precise (left panel) and

DNA repair genes play a pivotal role in the maintenance of genome integrity. Alterations of various DNA repair genes, either in gene sequence/structure or in gene expression, are frequently found in most of human malignancies. Since DNA repair activity is able to

and overall NHEJ repair (Fig. 4B).


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**21** 

*India* 

**Role of Radioprotectors in the Inhibition of DNA** 

Radiation has been considered an enigma to the general public and the use of radiation for therapeutic and other uses has always been associated with some skepticism. Presently, ionizing radiation is being used in a large number of therapeutic, industrial and other applications apart from for the generation of nuclear power, developing new varieties of high-yielding crops and enhancing storage period of food materials. Radiation treatment is an important therapeutic option for a number of malignancies, but its use is frequently limited due to adverse effects on normal tissues because it generates reactive oxygen species

body. The goal of most cancer treatments is to maximize the antineoplastic effect while

Due to the increased use of ionizing radiation in various aspects of human life, there is a need to develop an effective and non-toxic radioprotector. Radioprotectors are compounds that are designed to reduce the damage in normal tissues caused by radiation. These compounds are often antioxidants and must be present before or at the time of radiation for their effectiveness. Other agents, termed mitigators, may be used to minimize toxicity even after radiation has been delivered. Many natural and synthetic chemicals have been investigated in the recent past for their efficacy to protect against radiation-induced damage in biological systems (Maurya et al. 2006). Though a large number of compounds have been shown to be promising as radioprotectors in laboratory studies, few could pass the transition from bench to bedside. In fact, no radioprotective agent is now available, either alone or in combination to meet all the requisites of an ideal radioprotector. Amifostine is the only one that is currently in use having good radioprotection, even though there are reports about contraindications in some cases. Different radioprotectors offer protection to cellular molecules by different mechanisms (Maurya et al. 2006). Some of these compounds protect the target molecules because of their antioxidant mechanism by neutralizing the free radical, some enhances the cellular DNA repair (Maurya et al. 2005a; Maurya et al. 2005b), some modified the signaling pathways, some modulate the immune system and some

**1. Introduction** 

radicals (ROO.

(ROS) such as hydroxyl radical (·

minimizing harmful side effects for the patient.

contribute to a combination of all above mentioned mechanisms.

**Damage and Modulation of DNA Repair After** 

Dharmendra Kumar Maurya and Thomas Paul Asir Devasagayam

**Exposure to Gamma-Radiation** 

OH), superoxide radicals (O2-.), singlet oxygen and peroxyl

) in irradiated tissue that induce several pathophysiological changes in the

*Radiation Biology and Health Sciences Division, Bhabha Atomic Research Centre, Mumbai,* 

head and neck squamous cell carcinoma. *Clin Cancer Res*, Vol. 13, No.15 Pt 1, pp. 4386-4391.

