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M S Patil

Publications and source records attributed to M S Patil.

22 records · Page 2Linked to original sources

Nuclear matrix bound terminal deoxynucleotidyl transferase in rat thymus nuclei. I. A possible site for TdT mediated function.

Approximately 80% of the terminal deoxynucleotidyl transferase (TdT) in thymus glands from 3-4 week old rats was found to be localized in the nucleus and the remaining 20% in the cytosol. Following endogenous nuclease digestion of the thymus nuclei, 70-85% of the nuclear TdT could be removed by low salt and high salt extractions, whereas 15-30% of the enzyme remained tightly bound to the residual nuclear matrix. Low salt and high salt extracts of the nuclei contained a mixture of 58, 56, 45 and 44 kDa species of TdT whereas only 58 kDa species of the enzyme was found to be associated with the matrix. In addition to TdT, 20-25% of the nuclear DNA polymerase alpha was also tightly bound to the isolated nuclear matrix. These observations lead us to propose that besides being the site of DNA replication via-matrix bound replicational complexes [Van der Velden H.M.W. & Wanka F., Molecular Biology Reports 12 (1987): 69], nuclear matrix may also be the site of TdT mediated function and that matrix bound TdT and free TdT could be the functional and nonfunctional forms of the enzyme, respectively, in the thymus gland.

Affinity Labels↗

Nuclear matrix bound terminal deoxynucleotidyl transferase in rat thymus nuclei. II. Effect of ATP on free and matrix bound TdT.

Endogenous nuclease digestion of thymus nuclei from 3-4 week old rats followed by a step wise extraction with low salt, 0.5 M salt and 1 M salt removed approximately 70-85% of total nuclear terminal deoxynucleotidyl transferase (TdT) whereas approximately 15-30% of the enzyme remained tightly bound to the residual nuclear matrix. The cytoplasmic TdT as well as the bulk of nuclear TdT extracted in low salt and 0.5 M salt was found to be strongly inhibited at low concentration of ATP whereas matrix bound TdT and a significant portion of the enzyme in 1 M salt extract was completely insensitive to this nucleotide. The ATP resistant enzyme in the 1 M salt extract was unstable and slowly converted to ATP sensitive form upon prolonged preincubation on ice whereas under similar conditions it remained unaffected in the matrix bound form. These observations lead us to suggest that ATP resistant matrix bound TdT being capable of discriminating unnatural rNTPs against the natural dNTP substrates, may be the functionally organized form of the enzyme and that free TdT having lost the capability to distinguish between dNTP and rNTP may be the nonfunctional form of the enzyme in the thymus gland.

Adenosine Triphosphate↗

Effect of whole body hyperthermia on radiation therapy of transplanted fibrosarcoma in Swiss mice.

The exposure of normal mice to whole body hyperthermia (1 h WBH at 39 or 40 degrees C), 20 or 48 h prior to total body irradiation (TBI) with lethal doses of gamma-rays affords significant protection as assessed by survival. The radioprotective effect of WBH, as observed in normal mice, diminished in tumour bearing mice depending upon the size of tumour. Treatment of tumour bearing mice with mild WBH, 20 h prior to local irrradiation (LIR), did not protect the transplanted tumour against radiotherapy with a single dose of 20 Gy or fractionated dose (in five fractions) of 7.5 Gy on alternate days. In fact, mild WBH treatment enhanced the tumour regression and increased the mean survival time after fractionated dose therapy. However, the prior mild WBH was found to be ineffective in protecting normal tissue, as assessed by skin contraction after local irradiation (50 Gy). This indicates that mild WBH treatment given 20 h prior to local radiotherapy enhances fibrosarcoma tumour regression but cannot protect skin (normal tissue) against local irradiation. It appears that radioprotection of animals by WBH may be the consequence of its radioprotective effect on haemopoietic tissues mediated through certain cytokines. Perhaps WBH may not have a radioprotective effect on other tissues, as evident from skin contraction studies.

Animals↗