Method of identifying and assessing DNA euchromatin in biological cells for detecting disease, monitoring wellness, assessing bio-activity, and screening pharmacological agents

• US 20040092026A1
• Filed: 11/13/2002
• Published: 05/13/2004
• Est. Priority Date: 11/13/2002
• Status: Active Grant

• Current Assignee: G6 Science Corp.
• Original Assignee: G6 Science Corp.
• Inventors: Ferguson, Gary William
• Granted Patent: US 7,153,691 B2
• Time in Patent Office: Days
• US Class Current: 436/63
• CPC Class Codes: C12Q 1/68 involving nucleic acids
• CPC Class Codes: C12Q 1/6841 In situ hybridisation

Abstract

The present invention is a method of identifying and assessing DNA euchromatin in biological cells. The amount and/or distribution of DNA euchromatin generally relates to RNA/protein synthesis, which may change in certain conditions, such as disease, and/or cellular response to environmental and/or chemical agents. The present invention detects the potential presence of disease by assessing DNA euchromatin in ostensibly normal cells and similarly provides means to monitor and assess treatment response. The present invention may also be used to assess how application (or removal) of influences, such as environment, radiation, chemical agents, medications, herbs, vitamins, etc., interact to effect cells, establishing uses in monitoring wellness and pharmacological screening. Additional uses include assessing age related disorders, allergies, infections, Alzheimer'"'"'s disease, ‘Time of Death’, chronic fatigue syndrome, etc. The method may be advantageously applied to biological cells including micro-organisms, plants, animals, or cultured cells, alone or in conjunction with other markers.

Claims

1. A method of detecting disease based on the assessment of euchromatin, comprising

identifying euchromatin in a first set of biological cells,

assessing a variable relating to said euchromatin in said first set of biological cells, and

comparing said assessed variable to an expected value of said variable.

2. The method according to claim 1, wherein said identifying step comprises staining said euchromatin.

3. The method according to claim 2, wherein said staining step comprises using a DNA quantitative stain.

4. The method according to claim 3, wherein said DNA-quantitative stain is pararosaniline.

5. The method according to claim 3, wherein said DNA-quantitative stain is a thiazine derivative.

6. The method according to claim 5, wherein said thiazine derivative is thionin.

7. The method according to claim 2, wherein said staining step comprises Feulgen staining.

8. The method according to claim 7, further comprising preferentially hydrolyzing said euchromatin by performing said staining step at less than 25 degrees Celsius.

9. The method according to claim 8, wherein said staining step is performed at approximately 15 degrees Celsius.

10. The method according to claim 7, further comprising preferentially hydrolyzing said euchromatin for a time period between one minute and two hours.

11. The method according to claim 10, wherein said time period is between 15 minutes and two hours.

12. The method according to claim 7, further comprising performing said hydrolysis step with an acid in the range of 0.1 to 5.0 N.

13. The method according to claim 12, wherein said acid is hydrochloric acid.

14. The method according to claim 1, wherein said expected value is the normal value of said variable for said biological cells.

15. The method according to claim 14, wherein said expected value is stored in a database.

16. The method according to claim 1, wherein said expected value is determined from a reference data set detected in substantially healthy biological cells.

17. The method according to claim 16, wherein said expected value is stored in a database.

18. The method according to claim 1, wherein said assessed variable is the amount of said euchromatin.

19. The method according to claim 1, wherein said assessed variable is the distribution of said euchromatin.

20. The method according to claim 1, wherein said assessed variable is a cell response factor.

21. The method according to claim 20, wherein said cell response factor is determined from the amount of said euchromatin.

22. The method according to claim 21, wherein said cell response factor is determined from the distribution of said euchromatin.

23. The method according to claim 21, wherein said cell response factor is determined from the amount of said euchromatin and the distribution of said euchromatin.

24. The method according to claim 1, further comprising

assessing a second variable relating to said euchromatin in said nuclei of said first set of biological cells, and

comparing said second assessed variable to said expected value.

25. The method according to claim 1, wherein said subject is an animal.

26. The method according to claim 25, wherein said subject is a human.

27. The method according to claim 1, wherein said subject is a plant.

28. The method according to claim 1, wherein said subject is a micro-organism.

29. A method of assessing and monitoring wellness in a subject based on the assessment of euchromatin in biological cells, comprising

identifying euchromatin in a first set of biological cells,

assessing a variable relating to said euchromatin in said first set of biological cells,

establishing a basal level of said variable,

identifying euchromatin in a second set of biological cells,

assessing said variable relating to said euchromatin in said second set of biological cells, and

comparing said variable relating to said euchromatin in said second set of biological cells to said basal level.

30. The method according to claim 29, wherein said identifying steps comprise staining said euchromatin.

31. The method according to claim 30, wherein said staining step comprises using a DNA-quantitative stain.

32. The method according to claim 31, wherein said DNA-quantitative stain is pararosaniline.

33. The method according to claim 31, wherein said DNA-quantitative stain is a thiazine derivative.

34. The method according to claim 33, wherein said thiazine derivative is thionin.

35. The method according to claim 30, wherein said staining step comprises Feulgen staining.

36. The method according to claim 35, further comprising preferentially hydrolyzing said euchromatin by performing said hydrolysis step at less than 25 degrees Celsius.

37. The method according to claim 36, wherein said hydrolysis step is performed at approximately 15 degrees Celsius.

38. The method according to claim 35, further comprising preferentially hydrolyzing said euchromatin for a time period between one minute and two hours.

39. The method according to claim 38, wherein said time period is between 15 minutes and two hours.

40. The method according to claim 35, further comprising performing said hydrolysis step with an acid in the range of 0.1 to 5.0 N.

41. The method according to claim 40, wherein said acid is hydrochloric acid.

42. The method according to claim 29, wherein said assessed variable is the amount of said euchromatin.

43. The method according to claim 29, wherein said assessed variable is the distribution of said euchromatin.

44. The method according to claim 29, wherein said assessed variable is a cell response factor.

45. The method according to claim 44, wherein said cell response factor is determined from the amount of said euchromatin.

46. The method according to claim 44, wherein said cell response factor is determined from the distribution of said euchromatin.

47. The method according to claim 44, wherein said cell response factor is determined from the amount of said euchromatin and the distribution of said euchromatin.

48. The method according to claim 29, further comprising assessing a second variable relating to said euchromatin in said first set of biological cells.

49. The method according to claim 29, wherein said subject is an animal.

50. The method according to claim 49, wherein said subject is a human.

51. The method according to claim 29, wherein said subject is a plant.

52. The method according to claim 29, wherein said subject is a micro-organism.

53. A method of assessing and monitoring wellness in a subject based on the assessment of euchromatin in biological cells, comprising

identifying euchromatin in a first set of biological cells,

assessing a variable relating to said euchromatin in said first set of biological cells,

comparing said assessed variable to an expected value of said variable,

identifying euchromatin in a second set of biological cells,

assessing said variable relating to said euchromatin in said second set of biological cells, and

comparing said variable relating to said euchromatin in said second set of biological cells to said expected value of said variable.

54. The method according to claim 53, wherein said identifying steps comprise staining said euchromatin.

55. The method according to claim 54, wherein said staining step comprises using a DNA-quantitative stain.

56. The method according to claim 55, wherein said DNA-quantitative stain is pararosaniline.

57. The method according to claim 55, wherein said DNA-quantitative stain is a thiazine derivative.

58. The method according to claim 57, wherein said thiazine derivative is thionin.

59. The method according to claim 54, wherein said staining step comprises Feulgen staining.

60. The method according to claim 59, further comprising preferentially hydrolyzing said euchromatin by performing said staining step at less than 25 degrees Celsius.

61. The method according to claim 60, wherein said staining step is performed at approximately 15 degrees Celsius.

62. The method according to claim 59, further comprising preferentially hydrolyzing said euchromatin for a time period between one minute and two hours.

63. The method according to claim 62, wherein said time period is between 15 minutes and two hours.

64. The method according to claim 59, further comprising performing said staining step with an acid in the range of 0.1 to 5.0 N.

65. The method according to claim 64, wherein said acid is hydrochloric acid.

66. The method according to claim 53, wherein said assessed variable is the amount of said euchromatin.

67. The method according to claim 53, wherein said assessed variable is the distribution of said euchromatin.

68. The method according to claim 53, wherein said assessed variable is a cell response factor.

69. The method according to claim 68, wherein said cell response factor is determined from the amount of said euchromatin.

70. The method according to claim 68, wherein said cell response factor is determined from the distribution of said euchromatin.

71. The method according to claim 68, wherein said cell response factor is determined from the amount of said euchromatin and the distribution of said euchromatin.

72. The method according to claim 53, further comprising assessing a second variable relating to said euchromatin in said nuclei of said first set of biological cells.

73. The method according to claim 53, wherein said subject is an animal.

74. The method according to claim 73, wherein said subject is a human.

75. The method according to claim 53, wherein said subject is a plant.

76. The method according to claim 53, wherein said subject is a micro-organism.

77. A method of assessing response by a subject to exposure to an influence based on the assessment of euchromatin in biological cells, comprising

identifying euchromatin in a first set of biological cells,

assessing a variable relating to said euchromatin in first set of biological cells,

establishing a basal level of said variable,

exposing said subject to an influence,

identifying euchromatin in a second set of biological cells,

assessing said variable relating to said euchromatin in said second set of biological cells, and

comparing said variable relating to said euchromatin in said second set of biological cells to said basal level.

78. The method according to claim 77, wherein said identifying steps comprise staining said euchromatin.

79. The method according to claim 78, wherein said staining step comprises using a DNA-quantitative stain.

80. The method according to claim 79, wherein said DNA-quantitative stain is pararosaniline.

81. The method according to claim 79, wherein said DNA-quantitative stain is a thiazine derivative.

82. The method according to claim 81, wherein said thiazine derivative is thionin.

83. The method according to claim 78, wherein said staining step comprises Feulgen staining.

84. The method according to claim 83, further comprising preferentially hydrolyzing said euchromatin by performing said staining step at less than 25 degrees Celsius.

85. The method according to claim 84, wherein said staining step is performed at approximately 15 degrees Celsius.

86. The method according to claim 83, further comprising preferentially hydrolyzing said euchromatin for a time period between one minute and two hours.

87. The method according to claim 86, wherein said time period is between 15 minutes and two hours.

88. The method according to claim 83, further comprising performing said staining step with an acid in the range of 0.1 to 5.0 N.

89. The method according to claim 88, wherein said acid is hydrochloric acid.

90. The method according to claim 77, wherein said assessed variable is the amount of said euchromatin.

91. The method according to claim 77, wherein said assessed variable is the distribution of said euchromatin.

92. The method according to claim 77, wherein said assessed variable is a cell response factor.

93. The method according to claim 92, wherein said cell response factor is determined from the amount of said euchromatin.

94. The method according to claim 92, wherein said cell response factor is determined from the distribution of said euchromatin.

95. The method according to claim 92, wherein said cell response factor is determined from the amount of said euchromatin and the distribution of said euchromatin.

96. The method according to claim 77, further comprising assessing a second variable relating to said euchromatin in said nuclei of said first set of biological cells.

97. The method according to claim 77, wherein said subject is an animal.

98. The method according to claim 97, wherein said subject is a human.

99. The method according to claim 77, wherein said subject is a plant.

100. The method according to claim 77, wherein said subject is a micro-organ ism.

101. A method of assessing response by a subject to exposure to an influence based on the assessment of euchromatin in biological cells, comprising

identifying euchromatin in a first set of biological cells,

assessing a variable relating to said euchromatin in said first set of biological cells,

comparing said assessed variable to an expected value of said variable,

exposing said subject to an influence,

identifying euchromatin in said second set of biological cells,

assessing said variable relating to said euchromatin in said second set of biological cells, and

comparing said variable relating to said euchromatin in said second set of biological cells to said expected value of said variable.

102. The method according to claim 101, wherein said identifying steps comprise staining said euchromatin.

103. The method according to claim 102, wherein said staining step comprises using a DNA-quantitative stain.

104. The method according to claim 103, wherein said DNA-quantitative stain is pararosaniline.

105. The method according to claim 103, wherein said DNA-quantitative stain is a thiazine derivative.

106. The method according to claim 105, wherein said thiazine derivative is thionin.

107. The method according to claim 102, wherein said staining step comprises Feulgen staining.

108. The method according to claim 107, further comprising preferentially hydrolyzing said euchromatin by performing said staining step at less than 25 degrees Celsius.

109. The method according to claim 108, wherein said staining step is performed at approximately 15 degrees Celsius.

110. The method according to claim 107, further comprising preferentially hydrolyzing said euchromatin for a time period between one minute and two hours.

111. The method according to claim 110, wherein said time period is between 15 minutes and two hours.

112. The method according to claim 107, further comprising performing said staining step with an acid in the range of 0.1 to 5.0 N.

113. The method according to claim 112, wherein said acid is hydrochloric acid.

114. The method according to claim 101, wherein said assessed variable is the amount of said euchromatin.

115. The method according to claim 101, wherein said assessed variable is the distribution of said euchromatin.

116. The method according to claim 101, wherein said assessed variable is a cell response factor.

117. The method according to claim 116, wherein said cell response factor is determined from the amount of said euchromatin.

118. The method according to claim 116, wherein said cell response factor is determined from the distribution of said euchromatin.

119. The method according to claim 116, wherein said cell response factor is determined from the amount of said euchromatin and the distribution of said euchromatin.

120. The method according to claim 101, further comprising assessing a second variable relating to said euchromatin in said nuclei of said first set of biological cells.

121. The method according to claim 101, wherein said subject is an animal.

122. The method according to claim 121, wherein said subject is a human.

123. The method according to claim 101, wherein said subject is a plant.

124. The method according to claim 101, wherein said subject is a micro-organism.