WO1985001960A1 - Improved method of live blood cell analysis - Google Patents
Improved method of live blood cell analysis Download PDFInfo
- Publication number
- WO1985001960A1 WO1985001960A1 PCT/US1984/001673 US8401673W WO8501960A1 WO 1985001960 A1 WO1985001960 A1 WO 1985001960A1 US 8401673 W US8401673 W US 8401673W WO 8501960 A1 WO8501960 A1 WO 8501960A1
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- prescribing
- step comprises
- observation
- identification step
- identification
- Prior art date
Links
- 238000000034 method Methods 0.000 title claims abstract description 80
- 210000000601 blood cell Anatomy 0.000 title claims abstract description 19
- 238000004458 analytical method Methods 0.000 title abstract description 6
- 235000005911 diet Nutrition 0.000 claims abstract description 15
- 230000037213 diet Effects 0.000 claims abstract description 14
- 230000001575 pathological effect Effects 0.000 claims abstract description 7
- 229940088594 vitamin Drugs 0.000 claims abstract description 4
- 229930003231 vitamin Natural products 0.000 claims abstract description 4
- 239000011782 vitamin Substances 0.000 claims abstract description 4
- 210000004027 cell Anatomy 0.000 claims description 70
- 210000004369 blood Anatomy 0.000 claims description 42
- 239000008280 blood Substances 0.000 claims description 42
- 210000003743 erythrocyte Anatomy 0.000 claims description 28
- 210000000440 neutrophil Anatomy 0.000 claims description 20
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims description 16
- GVJHHUAWPYXKBD-UHFFFAOYSA-N (±)-α-Tocopherol Chemical compound OC1=C(C)C(C)=C2OC(CCCC(C)CCCC(C)CCCC(C)C)(C)CCC2=C1C GVJHHUAWPYXKBD-UHFFFAOYSA-N 0.000 claims description 15
- XEEYBQQBJWHFJM-UHFFFAOYSA-N Iron Chemical compound [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 claims description 14
- 206010039238 Rouleaux formation Diseases 0.000 claims description 14
- 235000019165 vitamin E Nutrition 0.000 claims description 14
- 239000011709 vitamin E Substances 0.000 claims description 14
- 230000002776 aggregation Effects 0.000 claims description 13
- 238000004220 aggregation Methods 0.000 claims description 13
- 235000019154 vitamin C Nutrition 0.000 claims description 13
- 239000011718 vitamin C Substances 0.000 claims description 13
- 235000019155 vitamin A Nutrition 0.000 claims description 12
- 239000011719 vitamin A Substances 0.000 claims description 12
- KKEBXNMGHUCPEZ-UHFFFAOYSA-N 4-phenyl-1-(2-sulfanylethyl)imidazolidin-2-one Chemical compound N1C(=O)N(CCS)CC1C1=CC=CC=C1 KKEBXNMGHUCPEZ-UHFFFAOYSA-N 0.000 claims description 11
- GHOKWGTUZJEAQD-ZETCQYMHSA-N (D)-(+)-Pantothenic acid Chemical compound OCC(C)(C)[C@@H](O)C(=O)NCCC(O)=O GHOKWGTUZJEAQD-ZETCQYMHSA-N 0.000 claims description 10
- HCHKCACWOHOZIP-UHFFFAOYSA-N Zinc Chemical compound [Zn] HCHKCACWOHOZIP-UHFFFAOYSA-N 0.000 claims description 9
- 210000001772 blood platelet Anatomy 0.000 claims description 9
- 210000003979 eosinophil Anatomy 0.000 claims description 9
- OVBPIULPVIDEAO-LBPRGKRZSA-N folic acid Chemical compound C=1N=C2NC(N)=NC(=O)C2=NC=1CNC1=CC=C(C(=O)N[C@@H](CCC(O)=O)C(O)=O)C=C1 OVBPIULPVIDEAO-LBPRGKRZSA-N 0.000 claims description 9
- PVNIIMVLHYAWGP-UHFFFAOYSA-N Niacin Chemical compound OC(=O)C1=CC=CN=C1 PVNIIMVLHYAWGP-UHFFFAOYSA-N 0.000 claims description 8
- 230000015572 biosynthetic process Effects 0.000 claims description 8
- 238000005755 formation reaction Methods 0.000 claims description 8
- 239000000463 material Substances 0.000 claims description 8
- 235000001968 nicotinic acid Nutrition 0.000 claims description 8
- 229960003512 nicotinic acid Drugs 0.000 claims description 8
- 239000011664 nicotinic acid Substances 0.000 claims description 8
- 239000013589 supplement Substances 0.000 claims description 8
- 239000011701 zinc Substances 0.000 claims description 8
- 229910052725 zinc Inorganic materials 0.000 claims description 8
- 235000016804 zinc Nutrition 0.000 claims description 8
- 229930003427 Vitamin E Natural products 0.000 claims description 7
- 235000019152 folic acid Nutrition 0.000 claims description 7
- 239000011724 folic acid Substances 0.000 claims description 7
- WIGCFUFOHFEKBI-UHFFFAOYSA-N gamma-tocopherol Natural products CC(C)CCCC(C)CCCC(C)CCCC1CCC2C(C)C(O)C(C)C(C)C2O1 WIGCFUFOHFEKBI-UHFFFAOYSA-N 0.000 claims description 7
- 229910052742 iron Inorganic materials 0.000 claims description 7
- 235000019156 vitamin B Nutrition 0.000 claims description 7
- 239000011720 vitamin B Substances 0.000 claims description 7
- 210000004430 acanthocyte Anatomy 0.000 claims description 6
- 239000013078 crystal Substances 0.000 claims description 6
- 235000021003 saturated fats Nutrition 0.000 claims description 6
- 229940046009 vitamin E Drugs 0.000 claims description 6
- GHOKWGTUZJEAQD-UHFFFAOYSA-N Chick antidermatitis factor Natural products OCC(C)(C)C(O)C(=O)NCCC(O)=O GHOKWGTUZJEAQD-UHFFFAOYSA-N 0.000 claims description 5
- 239000003963 antioxidant agent Substances 0.000 claims description 5
- 235000006708 antioxidants Nutrition 0.000 claims description 5
- FDJOLVPMNUYSCM-WZHZPDAFSA-L cobalt(3+);[(2r,3s,4r,5s)-5-(5,6-dimethylbenzimidazol-1-yl)-4-hydroxy-2-(hydroxymethyl)oxolan-3-yl] [(2r)-1-[3-[(1r,2r,3r,4z,7s,9z,12s,13s,14z,17s,18s,19r)-2,13,18-tris(2-amino-2-oxoethyl)-7,12,17-tris(3-amino-3-oxopropyl)-3,5,8,8,13,15,18,19-octamethyl-2 Chemical compound [Co+3].N#[C-].N([C@@H]([C@]1(C)[N-]\C([C@H]([C@@]1(CC(N)=O)C)CCC(N)=O)=C(\C)/C1=N/C([C@H]([C@@]1(CC(N)=O)C)CCC(N)=O)=C\C1=N\C([C@H](C1(C)C)CCC(N)=O)=C/1C)[C@@H]2CC(N)=O)=C\1[C@]2(C)CCC(=O)NC[C@@H](C)OP([O-])(=O)O[C@H]1[C@@H](O)[C@@H](N2C3=CC(C)=C(C)C=C3N=C2)O[C@@H]1CO FDJOLVPMNUYSCM-WZHZPDAFSA-L 0.000 claims description 5
- 230000000694 effects Effects 0.000 claims description 5
- 229940014144 folate Drugs 0.000 claims description 5
- 229940055726 pantothenic acid Drugs 0.000 claims description 5
- 235000019161 pantothenic acid Nutrition 0.000 claims description 5
- 239000011713 pantothenic acid Substances 0.000 claims description 5
- YBJHBAHKTGYVGT-ZKWXMUAHSA-N (+)-Biotin Chemical compound N1C(=O)N[C@@H]2[C@H](CCCCC(=O)O)SC[C@@H]21 YBJHBAHKTGYVGT-ZKWXMUAHSA-N 0.000 claims description 4
- CIWBSHSKHKDKBQ-JLAZNSOCSA-N Ascorbic acid Chemical compound OC[C@H](O)[C@H]1OC(=O)C(O)=C1O CIWBSHSKHKDKBQ-JLAZNSOCSA-N 0.000 claims description 4
- 210000002652 macrocyte Anatomy 0.000 claims description 4
- 210000001938 protoplast Anatomy 0.000 claims description 4
- -1 thymus Chemical compound 0.000 claims description 4
- 210000001541 thymus gland Anatomy 0.000 claims description 3
- IIZPXYDJLKNOIY-JXPKJXOSSA-N 1-palmitoyl-2-arachidonoyl-sn-glycero-3-phosphocholine Chemical compound CCCCCCCCCCCCCCCC(=O)OC[C@H](COP([O-])(=O)OCC[N+](C)(C)C)OC(=O)CCC\C=C/C\C=C/C\C=C/C\C=C/CCCCC IIZPXYDJLKNOIY-JXPKJXOSSA-N 0.000 claims description 2
- SQDAZGGFXASXDW-UHFFFAOYSA-N 5-bromo-2-(trifluoromethoxy)pyridine Chemical compound FC(F)(F)OC1=CC=C(Br)C=N1 SQDAZGGFXASXDW-UHFFFAOYSA-N 0.000 claims description 2
- 229920001287 Chondroitin sulfate Polymers 0.000 claims description 2
- ZZZCUOFIHGPKAK-UHFFFAOYSA-N D-erythro-ascorbic acid Natural products OCC1OC(=O)C(O)=C1O ZZZCUOFIHGPKAK-UHFFFAOYSA-N 0.000 claims description 2
- FFDGPVCHZBVARC-UHFFFAOYSA-N N,N-dimethylglycine Chemical compound CN(C)CC(O)=O FFDGPVCHZBVARC-UHFFFAOYSA-N 0.000 claims description 2
- OVBPIULPVIDEAO-UHFFFAOYSA-N N-Pteroyl-L-glutaminsaeure Natural products C=1N=C2NC(N)=NC(=O)C2=NC=1CNC1=CC=C(C(=O)NC(CCC(O)=O)C(O)=O)C=C1 OVBPIULPVIDEAO-UHFFFAOYSA-N 0.000 claims description 2
- 229930003268 Vitamin C Natural products 0.000 claims description 2
- OENHQHLEOONYIE-UKMVMLAPSA-N all-trans beta-carotene Natural products CC=1CCCC(C)(C)C=1/C=C/C(/C)=C/C=C/C(/C)=C/C=C/C=C(C)C=CC=C(C)C=CC1=C(C)CCCC1(C)C OENHQHLEOONYIE-UKMVMLAPSA-N 0.000 claims description 2
- TUPZEYHYWIEDIH-WAIFQNFQSA-N beta-carotene Natural products CC(=C/C=C/C=C(C)/C=C/C=C(C)/C=C/C1=C(C)CCCC1(C)C)C=CC=C(/C)C=CC2=CCCCC2(C)C TUPZEYHYWIEDIH-WAIFQNFQSA-N 0.000 claims description 2
- 235000013734 beta-carotene Nutrition 0.000 claims description 2
- 239000011648 beta-carotene Substances 0.000 claims description 2
- 229960002747 betacarotene Drugs 0.000 claims description 2
- 229960002685 biotin Drugs 0.000 claims description 2
- 235000020958 biotin Nutrition 0.000 claims description 2
- 239000011616 biotin Substances 0.000 claims description 2
- 229940059329 chondroitin sulfate Drugs 0.000 claims description 2
- 239000003026 cod liver oil Substances 0.000 claims description 2
- 235000012716 cod liver oil Nutrition 0.000 claims description 2
- 108700003601 dimethylglycine Proteins 0.000 claims description 2
- 229960000304 folic acid Drugs 0.000 claims description 2
- 210000003714 granulocyte Anatomy 0.000 claims description 2
- 239000000787 lecithin Substances 0.000 claims description 2
- 229940067606 lecithin Drugs 0.000 claims description 2
- 235000010445 lecithin Nutrition 0.000 claims description 2
- 244000005700 microbiome Species 0.000 claims description 2
- 238000000386 microscopy Methods 0.000 claims description 2
- 235000013343 vitamin Nutrition 0.000 claims description 2
- OENHQHLEOONYIE-JLTXGRSLSA-N β-Carotene Chemical compound CC=1CCCC(C)(C)C=1\C=C\C(\C)=C\C=C\C(\C)=C\C=C\C=C(/C)\C=C\C=C(/C)\C=C\C1=C(C)CCCC1(C)C OENHQHLEOONYIE-JLTXGRSLSA-N 0.000 claims description 2
- FPIPGXGPPPQFEQ-UHFFFAOYSA-N 13-cis retinol Natural products OCC=C(C)C=CC=C(C)C=CC1=C(C)CCCC1(C)C FPIPGXGPPPQFEQ-UHFFFAOYSA-N 0.000 claims 1
- 241000186660 Lactobacillus Species 0.000 claims 1
- FPIPGXGPPPQFEQ-BOOMUCAASA-N Vitamin A Natural products OC/C=C(/C)\C=C\C=C(\C)/C=C/C1=C(C)CCCC1(C)C FPIPGXGPPPQFEQ-BOOMUCAASA-N 0.000 claims 1
- 229930003270 Vitamin B Natural products 0.000 claims 1
- FPIPGXGPPPQFEQ-OVSJKPMPSA-N all-trans-retinol Chemical compound OC\C=C(/C)\C=C\C=C(/C)\C=C\C1=C(C)CCCC1(C)C FPIPGXGPPPQFEQ-OVSJKPMPSA-N 0.000 claims 1
- 229940039696 lactobacillus Drugs 0.000 claims 1
- 229940045997 vitamin a Drugs 0.000 claims 1
- 238000002360 preparation method Methods 0.000 abstract description 53
- 239000005445 natural material Substances 0.000 abstract description 4
- 150000003722 vitamin derivatives Chemical class 0.000 abstract description 3
- 230000001413 cellular effect Effects 0.000 abstract description 2
- 229940029985 mineral supplement Drugs 0.000 abstract description 2
- 235000020786 mineral supplement Nutrition 0.000 abstract description 2
- 230000008068 pathophysiological alteration Effects 0.000 abstract description 2
- 235000019195 vitamin supplement Nutrition 0.000 abstract description 2
- 230000004075 alteration Effects 0.000 abstract 1
- 238000010186 staining Methods 0.000 description 11
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 10
- 208000015181 infectious disease Diseases 0.000 description 9
- 206010036790 Productive cough Diseases 0.000 description 8
- 239000003921 oil Substances 0.000 description 8
- 235000019198 oils Nutrition 0.000 description 8
- 210000003802 sputum Anatomy 0.000 description 8
- 208000024794 sputum Diseases 0.000 description 8
- 238000011156 evaluation Methods 0.000 description 7
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 6
- 238000003745 diagnosis Methods 0.000 description 6
- 239000003925 fat Substances 0.000 description 6
- 235000019197 fats Nutrition 0.000 description 6
- 230000002489 hematologic effect Effects 0.000 description 6
- 229910052760 oxygen Inorganic materials 0.000 description 6
- 239000001301 oxygen Substances 0.000 description 6
- 238000012360 testing method Methods 0.000 description 6
- 238000002560 therapeutic procedure Methods 0.000 description 6
- 241000894006 Bacteria Species 0.000 description 5
- 230000005856 abnormality Effects 0.000 description 5
- 201000010099 disease Diseases 0.000 description 5
- 208000035475 disorder Diseases 0.000 description 5
- 208000010110 spontaneous platelet aggregation Diseases 0.000 description 5
- 206010002065 Anaemia megaloblastic Diseases 0.000 description 4
- 208000000682 Megaloblastic Anemia Diseases 0.000 description 4
- 206010028980 Neoplasm Diseases 0.000 description 4
- 208000006011 Stroke Diseases 0.000 description 4
- 230000002159 abnormal effect Effects 0.000 description 4
- 208000007502 anemia Diseases 0.000 description 4
- 231100001016 megaloblastic anemia Toxicity 0.000 description 4
- 235000016709 nutrition Nutrition 0.000 description 4
- 206010002536 Anisocytosis Diseases 0.000 description 3
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 3
- 230000001594 aberrant effect Effects 0.000 description 3
- 239000003242 anti bacterial agent Substances 0.000 description 3
- 229940088710 antibiotic agent Drugs 0.000 description 3
- 201000011510 cancer Diseases 0.000 description 3
- 238000001446 dark-field microscopy Methods 0.000 description 3
- 230000007812 deficiency Effects 0.000 description 3
- 238000001514 detection method Methods 0.000 description 3
- 238000011161 development Methods 0.000 description 3
- 230000018109 developmental process Effects 0.000 description 3
- 229940079593 drug Drugs 0.000 description 3
- 239000003814 drug Substances 0.000 description 3
- 230000002538 fungal effect Effects 0.000 description 3
- 229910052500 inorganic mineral Inorganic materials 0.000 description 3
- 210000004185 liver Anatomy 0.000 description 3
- 238000004519 manufacturing process Methods 0.000 description 3
- 235000012054 meals Nutrition 0.000 description 3
- 239000011707 mineral Substances 0.000 description 3
- 235000010755 mineral Nutrition 0.000 description 3
- 230000000877 morphologic effect Effects 0.000 description 3
- 230000001991 pathophysiological effect Effects 0.000 description 3
- 238000011160 research Methods 0.000 description 3
- 210000002700 urine Anatomy 0.000 description 3
- GVJHHUAWPYXKBD-IEOSBIPESA-N α-tocopherol Chemical compound OC1=C(C)C(C)=C2O[C@@](CCC[C@H](C)CCC[C@H](C)CCCC(C)C)(C)CCC2=C1C GVJHHUAWPYXKBD-IEOSBIPESA-N 0.000 description 3
- 208000019838 Blood disease Diseases 0.000 description 2
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 description 2
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 2
- 102000008946 Fibrinogen Human genes 0.000 description 2
- 108010049003 Fibrinogen Proteins 0.000 description 2
- 206010016880 Folate deficiency Diseases 0.000 description 2
- 208000010188 Folic Acid Deficiency Diseases 0.000 description 2
- 206010018910 Haemolysis Diseases 0.000 description 2
- 206010020772 Hypertension Diseases 0.000 description 2
- 208000006595 Necrotizing Ulcerative Gingivitis Diseases 0.000 description 2
- 206010035774 Poikilocytosis Diseases 0.000 description 2
- 239000003146 anticoagulant agent Substances 0.000 description 2
- 239000012153 distilled water Substances 0.000 description 2
- 238000001035 drying Methods 0.000 description 2
- 229940012952 fibrinogen Drugs 0.000 description 2
- 235000013305 food Nutrition 0.000 description 2
- 230000006870 function Effects 0.000 description 2
- 208000014951 hematologic disease Diseases 0.000 description 2
- 208000018706 hematopoietic system disease Diseases 0.000 description 2
- 230000008588 hemolysis Effects 0.000 description 2
- 208000007475 hemolytic anemia Diseases 0.000 description 2
- 208000006278 hypochromic anemia Diseases 0.000 description 2
- 238000005286 illumination Methods 0.000 description 2
- XVARCVCWNFACQC-RKQHYHRCSA-N indican Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1OC1=CNC2=CC=CC=C12 XVARCVCWNFACQC-RKQHYHRCSA-N 0.000 description 2
- 235000004213 low-fat Nutrition 0.000 description 2
- 229920002521 macromolecule Polymers 0.000 description 2
- 230000004660 morphological change Effects 0.000 description 2
- 208000015122 neurodegenerative disease Diseases 0.000 description 2
- 230000003448 neutrophilic effect Effects 0.000 description 2
- 230000007505 plaque formation Effects 0.000 description 2
- 102000004169 proteins and genes Human genes 0.000 description 2
- 108090000623 proteins and genes Proteins 0.000 description 2
- LXNHXLLTXMVWPM-UHFFFAOYSA-N pyridoxine Chemical compound CC1=NC=C(CO)C(CO)=C1O LXNHXLLTXMVWPM-UHFFFAOYSA-N 0.000 description 2
- 230000007480 spreading Effects 0.000 description 2
- 238000003892 spreading Methods 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 230000008685 targeting Effects 0.000 description 2
- 231100000419 toxicity Toxicity 0.000 description 2
- 230000001988 toxicity Effects 0.000 description 2
- 229960005486 vaccine Drugs 0.000 description 2
- 235000019158 vitamin B6 Nutrition 0.000 description 2
- 239000011726 vitamin B6 Substances 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- 241000203809 Actinomycetales Species 0.000 description 1
- 241000251468 Actinopterygii Species 0.000 description 1
- 102000057234 Acyl transferases Human genes 0.000 description 1
- 108700016155 Acyl transferases Proteins 0.000 description 1
- 241000234282 Allium Species 0.000 description 1
- 235000002732 Allium cepa var. cepa Nutrition 0.000 description 1
- 240000002234 Allium sativum Species 0.000 description 1
- 102100033312 Alpha-2-macroglobulin Human genes 0.000 description 1
- 206010003210 Arteriosclerosis Diseases 0.000 description 1
- 201000001320 Atherosclerosis Diseases 0.000 description 1
- 206010004053 Bacterial toxaemia Diseases 0.000 description 1
- 206010009208 Cirrhosis alcoholic Diseases 0.000 description 1
- 229920002307 Dextran Polymers 0.000 description 1
- 102000009123 Fibrin Human genes 0.000 description 1
- 108010073385 Fibrin Proteins 0.000 description 1
- BWGVNKXGVNDBDI-UHFFFAOYSA-N Fibrin monomer Chemical compound CNC(=O)CNC(=O)CN BWGVNKXGVNDBDI-UHFFFAOYSA-N 0.000 description 1
- 208000034502 Haemoglobin C disease Diseases 0.000 description 1
- 108010054147 Hemoglobins Proteins 0.000 description 1
- 102000001554 Hemoglobins Human genes 0.000 description 1
- SQUHHTBVTRBESD-UHFFFAOYSA-N Hexa-Ac-myo-Inositol Natural products CC(=O)OC1C(OC(C)=O)C(OC(C)=O)C(OC(C)=O)C(OC(C)=O)C1OC(C)=O SQUHHTBVTRBESD-UHFFFAOYSA-N 0.000 description 1
- 206010020751 Hypersensitivity Diseases 0.000 description 1
- 206010022971 Iron Deficiencies Diseases 0.000 description 1
- 208000015710 Iron-Deficiency Anemia Diseases 0.000 description 1
- 206010023129 Jaundice cholestatic Diseases 0.000 description 1
- FFEARJCKVFRZRR-BYPYZUCNSA-N L-methionine Chemical compound CSCC[C@H](N)C(O)=O FFEARJCKVFRZRR-BYPYZUCNSA-N 0.000 description 1
- 206010069698 Langerhans' cell histiocytosis Diseases 0.000 description 1
- FYYHWMGAXLPEAU-UHFFFAOYSA-N Magnesium Chemical compound [Mg] FYYHWMGAXLPEAU-UHFFFAOYSA-N 0.000 description 1
- 206010027540 Microcytosis Diseases 0.000 description 1
- 206010028665 Myxoedema Diseases 0.000 description 1
- 206010055670 Necrotising ulcerative gingivostomatitis Diseases 0.000 description 1
- 201000005267 Obstructive Jaundice Diseases 0.000 description 1
- 102000003992 Peroxidases Human genes 0.000 description 1
- 201000007100 Pharyngitis Diseases 0.000 description 1
- 241000255969 Pieris brassicae Species 0.000 description 1
- 108010015078 Pregnancy-Associated alpha 2-Macroglobulins Proteins 0.000 description 1
- 206010039710 Scleroderma Diseases 0.000 description 1
- 208000013222 Toxemia Diseases 0.000 description 1
- 208000032109 Transient ischaemic attack Diseases 0.000 description 1
- 241000589886 Treponema Species 0.000 description 1
- 208000022440 X-linked sideroblastic anemia 1 Diseases 0.000 description 1
- 208000004622 abetalipoproteinemia Diseases 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 208000010002 alcoholic liver cirrhosis Diseases 0.000 description 1
- 208000026935 allergic disease Diseases 0.000 description 1
- 230000007815 allergy Effects 0.000 description 1
- 239000001068 allium cepa oil Substances 0.000 description 1
- 229940087168 alpha tocopherol Drugs 0.000 description 1
- 230000002744 anti-aggregatory effect Effects 0.000 description 1
- 229940127219 anticoagulant drug Drugs 0.000 description 1
- 230000003078 antioxidant effect Effects 0.000 description 1
- 208000011775 arteriosclerosis disease Diseases 0.000 description 1
- POJOORKDYOPQLS-UHFFFAOYSA-L barium(2+) 5-chloro-2-[(2-hydroxynaphthalen-1-yl)diazenyl]-4-methylbenzenesulfonate Chemical compound [Ba+2].C1=C(Cl)C(C)=CC(N=NC=2C3=CC=CC=C3C=CC=2O)=C1S([O-])(=O)=O.C1=C(Cl)C(C)=CC(N=NC=2C3=CC=CC=C3C=CC=2O)=C1S([O-])(=O)=O POJOORKDYOPQLS-UHFFFAOYSA-L 0.000 description 1
- 238000004159 blood analysis Methods 0.000 description 1
- 210000001124 body fluid Anatomy 0.000 description 1
- 239000010839 body fluid Substances 0.000 description 1
- 210000001185 bone marrow Anatomy 0.000 description 1
- 244000309464 bull Species 0.000 description 1
- 239000001569 carbon dioxide Substances 0.000 description 1
- 229910002092 carbon dioxide Inorganic materials 0.000 description 1
- 150000003943 catecholamines Chemical class 0.000 description 1
- 210000000170 cell membrane Anatomy 0.000 description 1
- 210000002421 cell wall Anatomy 0.000 description 1
- GTRGJJDVSJFNTE-UHFFFAOYSA-N chembl2009633 Chemical compound OC1=CC=C2C=C(S(O)(=O)=O)C=CC2=C1N=NC1=CC=CC=C1 GTRGJJDVSJFNTE-UHFFFAOYSA-N 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 229960001231 choline Drugs 0.000 description 1
- OEYIOHPDSNJKLS-UHFFFAOYSA-N choline Chemical compound C[N+](C)(C)CCO OEYIOHPDSNJKLS-UHFFFAOYSA-N 0.000 description 1
- 230000001684 chronic effect Effects 0.000 description 1
- 239000000470 constituent Substances 0.000 description 1
- 208000029078 coronary artery disease Diseases 0.000 description 1
- 230000000093 cytochemical effect Effects 0.000 description 1
- 230000002380 cytological effect Effects 0.000 description 1
- 230000006378 damage Effects 0.000 description 1
- 230000007123 defense Effects 0.000 description 1
- 230000000378 dietary effect Effects 0.000 description 1
- 102000038379 digestive enzymes Human genes 0.000 description 1
- 108091007734 digestive enzymes Proteins 0.000 description 1
- 230000003292 diminished effect Effects 0.000 description 1
- 231100000676 disease causative agent Toxicity 0.000 description 1
- 238000001493 electron microscopy Methods 0.000 description 1
- 238000004945 emulsification Methods 0.000 description 1
- 230000002708 enhancing effect Effects 0.000 description 1
- 208000003401 eosinophilic granuloma Diseases 0.000 description 1
- 229950003499 fibrin Drugs 0.000 description 1
- 235000019688 fish Nutrition 0.000 description 1
- 239000012530 fluid Substances 0.000 description 1
- ZZUFCTLCJUWOSV-UHFFFAOYSA-N furosemide Chemical compound C1=C(Cl)C(S(=O)(=O)N)=CC(C(O)=O)=C1NCC1=CC=CO1 ZZUFCTLCJUWOSV-UHFFFAOYSA-N 0.000 description 1
- 235000004611 garlic Nutrition 0.000 description 1
- 239000010647 garlic oil Substances 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- 239000008187 granular material Substances 0.000 description 1
- 208000019622 heart disease Diseases 0.000 description 1
- 208000006454 hepatitis Diseases 0.000 description 1
- 231100000283 hepatitis Toxicity 0.000 description 1
- 230000001744 histochemical effect Effects 0.000 description 1
- 235000011167 hydrochloric acid Nutrition 0.000 description 1
- 208000009300 hypochromic microcytic anemia Diseases 0.000 description 1
- 230000001900 immune effect Effects 0.000 description 1
- BXFFHSIDQOFMLE-UHFFFAOYSA-N indoxyl sulfate Natural products C1=CC=C2C(OS(=O)(=O)O)=CNC2=C1 BXFFHSIDQOFMLE-UHFFFAOYSA-N 0.000 description 1
- XVARCVCWNFACQC-UHFFFAOYSA-N indoxyl-beta-D-glucopyranoside Natural products OC1C(O)C(O)C(CO)OC1OC1=CNC2=CC=CC=C12 XVARCVCWNFACQC-UHFFFAOYSA-N 0.000 description 1
- 230000001939 inductive effect Effects 0.000 description 1
- 229960000367 inositol Drugs 0.000 description 1
- CDAISMWEOUEBRE-GPIVLXJGSA-N inositol Chemical compound O[C@H]1[C@H](O)[C@@H](O)[C@H](O)[C@H](O)[C@@H]1O CDAISMWEOUEBRE-GPIVLXJGSA-N 0.000 description 1
- 238000007689 inspection Methods 0.000 description 1
- 210000000265 leukocyte Anatomy 0.000 description 1
- 235000021388 linseed oil Nutrition 0.000 description 1
- 239000000944 linseed oil Substances 0.000 description 1
- 230000037356 lipid metabolism Effects 0.000 description 1
- 230000003212 lipotrophic effect Effects 0.000 description 1
- 230000003908 liver function Effects 0.000 description 1
- 239000011777 magnesium Substances 0.000 description 1
- 229910052749 magnesium Inorganic materials 0.000 description 1
- 229930182817 methionine Natural products 0.000 description 1
- 229960004452 methionine Drugs 0.000 description 1
- 244000000010 microbial pathogen Species 0.000 description 1
- 238000007431 microscopic evaluation Methods 0.000 description 1
- 208000003786 myxedema Diseases 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 230000004792 oxidative damage Effects 0.000 description 1
- 230000000242 pagocytic effect Effects 0.000 description 1
- 230000003071 parasitic effect Effects 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 230000007170 pathology Effects 0.000 description 1
- 230000004963 pathophysiological condition Effects 0.000 description 1
- 108040007629 peroxidase activity proteins Proteins 0.000 description 1
- 230000036195 photohemolysis Effects 0.000 description 1
- RADKZDMFGJYCBB-UHFFFAOYSA-N pyridoxal hydrochloride Natural products CC1=NC=C(CO)C(C=O)=C1O RADKZDMFGJYCBB-UHFFFAOYSA-N 0.000 description 1
- 206010039073 rheumatoid arthritis Diseases 0.000 description 1
- 238000004626 scanning electron microscopy Methods 0.000 description 1
- CDAISMWEOUEBRE-UHFFFAOYSA-N scyllo-inosotol Natural products OC1C(O)C(O)C(O)C(O)C1O CDAISMWEOUEBRE-UHFFFAOYSA-N 0.000 description 1
- 238000004062 sedimentation Methods 0.000 description 1
- 230000011218 segmentation Effects 0.000 description 1
- 208000007056 sickle cell anemia Diseases 0.000 description 1
- 239000000243 solution Substances 0.000 description 1
- 239000012192 staining solution Substances 0.000 description 1
- 239000011550 stock solution Substances 0.000 description 1
- 208000003265 stomatitis Diseases 0.000 description 1
- 230000001502 supplementing effect Effects 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- 210000001519 tissue Anatomy 0.000 description 1
- 229960000984 tocofersolan Drugs 0.000 description 1
- 229930003799 tocopherol Natural products 0.000 description 1
- 229960001295 tocopherol Drugs 0.000 description 1
- 239000011732 tocopherol Substances 0.000 description 1
- 235000010384 tocopherol Nutrition 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 201000010875 transient cerebral ischemia Diseases 0.000 description 1
- 208000001072 type 2 diabetes mellitus Diseases 0.000 description 1
- 235000021081 unsaturated fats Nutrition 0.000 description 1
- 235000013311 vegetables Nutrition 0.000 description 1
- 230000035899 viability Effects 0.000 description 1
- 230000000007 visual effect Effects 0.000 description 1
- 238000011179 visual inspection Methods 0.000 description 1
- 229940011671 vitamin b6 Drugs 0.000 description 1
- 235000013618 yogurt Nutrition 0.000 description 1
- 235000004835 α-tocopherol Nutrition 0.000 description 1
- 239000002076 α-tocopherol Substances 0.000 description 1
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
- G01N15/10—Investigating individual particles
- G01N15/14—Optical investigation techniques, e.g. flow cytometry
- G01N15/1429—Signal processing
- G01N15/1433—Signal processing using image recognition
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
- G01N15/01—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials specially adapted for biological cells, e.g. blood cells
- G01N2015/012—Red blood cells
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
- G01N15/10—Investigating individual particles
- G01N15/14—Optical investigation techniques, e.g. flow cytometry
- G01N2015/1493—Particle size
- G01N2015/1495—Deformation of particles
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
- G01N15/10—Investigating individual particles
- G01N15/14—Optical investigation techniques, e.g. flow cytometry
- G01N2015/1497—Particle shape
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/483—Physical analysis of biological material
- G01N33/487—Physical analysis of biological material of liquid biological material
- G01N33/49—Blood
Definitions
- the thickness of the smear is a function of the speed with which the coverslip is moved across the slide.
- the smearing technique must be such that in the final film, the cells generally lie separated side by side with some cells being aggregated in small rouleaux. Smears that are made too thick usually result in overstaining and some clumping, making microscopic analysis of the cells practically impossible.
- the acquisition of a proper smearing technique is essential to clinical hematology and can only be acquired by practical experience.
- the smears must then be dried rapidly in order to preserve the shape of the majority of the cells; drying may be' accomplished by the rapid movement of the slide, through the air, the use of previously heated slides, or the preparation of smears under an infrared lamp.
- OMP WIP preparation is subjected to certain standard staining procedures.
- the most widely used staining procedure Pappenheim
- the staining technique must be adapted to the requirements of the individual laboratory based on the pH of the distilled water and the frequency of use and replacement of the staining stock solutions.
- Another common staining procedure is to use commercially available Wright's Stain.
- Other special staining and cytochemical (histochemical) methods are also available.
- the preparation of a slide requires several hours, and must be accomplished in a laboratory established for that purpose. Therefore, the microscopic examination and evaluation must necessarily occur sometime after the blood has been withdrawn from the patient. Because of the time and special facilities necessary to prepare the slides and perform the microscopic examination, generally the attending physician does not perform the slide evaluation and, further, the results are generally not available until the following day and then presented in such a manner, typically by telephone, as to be useful only to the attending physician and, therefore, have less effect on the patient who does not have an opportunity to view the blood preparations.
- the present invention provides a method of detecting, diagnosing, and prescribing treatment for a variety of pathological conditions that are generally manifested in hematological abnormalities.
- the invention provides a method by which live blood cells can be subjected to microscopic examination immediately after blood has been withdrawn from the patient and dispensing with the expensive and time-consuming necessity of- preparing a standard blood smear slide.
- the method utilizes a microscope with a dark field condenser and with a video camera attachment thereby permitting simultaneous viewing of the live blood cell preparation by both the attending physician and the patient. The method thus permits the attending physician or techician to evaluate immediately the blood preparation, and provides an immediate feedback to the patient on the evaluation and diagnosis of the blood, while offering a visual presentation of the cells
- OMPI that the attending physican may use to actually show the patient any abnormalities observed.
- Figures 1-23 are photomicrographs of live blood cell preparations as viewed under conventional dark field illumination using a 100X oil emersion objective, except for Figure 3 which utilizes a 40X objective;
- Figures 24 and 25 are photomicrographs of sputum preparations viewed under conventional dark field illumination, utilizing a 100X oil emersion objective.
- DETAILED DESCRIPTION OF THE INVENTION The basis of the present invention described is the detection and identification of pathophysiologically altered cells in a live state. Although these morphological changes and inclusions in cell preparations can be visualized using standard blood smear preparations, the advent of phase contrast, dark field, fluorescent, electron, and scanning electron microscopy have made the identification of many morphological features of cells possible without the need for the more complicated and difficult slide preparation and staining procedures.
- the invention uses dark field microscopy which profoundly enhances morphological characteristics of living cells such as size, shape, and type of cells, as well as major inclusions within the cell and abnormal constituents of the surrounding fluid.
- the invention has compiled a series of the more common pathophysiological states of living cells that can be identified by morphological changes or abnormal inclusions and has reinterpreted these findings to diagnose a pathological state or condition of the body manifested by the pathophysiological state of the living cells. More importantly, the evaluation of the live cell preparation is done within minutes after having withdrawn the blood from the patient and is accomplished in the presence of the patient such that the attending physician or technician may point out the differences between the normal state and any observed pathophysiological state of the cells.
- the invention provides a blood examination procedure that may also be used by the attending physician to clearly illustrate to the patient abnormalities that may occur in the blood.
- the patient not only receives diagnostic information that can be readily understood, but the entire procedure of drawing the patient's blood, evaluation of the blood preparation, diagnosis, explanation of the diagnostic results to the patient, and the prescribing of a treatment for any abnormal conditions that may be found, may be accomplished within a short period of time such as less than one hour.
- the instrumentation used in the microscopic examination of the living cell preparation includes a standard bright field microscope (Bausch and Lomb, Model
- DRT3Z1H that has been equipped with 10X, 20X, 40X, and
- the microscope is additionally configured with a conventional video camera that is mounted on the microscope, the image of which-is displayed on a standard television set.
- the procedure involved for obtaining a blood specimen and preparing the live blood cell preparation involves obtaining a sample of blood from a patient using a standard finger prick.
- the patient's finger is cleaned well with an alcohol swab then wiped dry with lint-free tissue.
- the center of a precleaned glass slide is touched gently to the drop of blood taking care not to touch any portion of the finger.
- the slide is subsequently rotated such that the blood sample is now positioned on the top surface of the slide and a standard coverslip (22x40) is then placed on the slide over the sample of blood.
- a drop of emersion oil is placed between the slide and the condenser and a 10X objective is used to scan the blood preparation to ascertain that portion of the blood preparation where the blood appears the thinnest and the cells most evenly distributed.
- a drop of oil is then placed on the coverslip and the 100X oil emersion objective is used for the evaluation of the blood cell preparation, except when thrombocyte aggregations are to be viewed, wherein the 40X objection is occasionally used (without oil).
- the procedure described above refers to the preparation of a blood sample and that the procedure can be used for the preparation of other samples such as sputum and urine.
- the term blood sample shall be defined to include blood, sputum, urine, and other body fluid specimens.
- the live blood cell preparation is then searched for abnormalities, each of which is separately discussed below:
- red blood cells have a general biconcave shape. Normally, these red blood cells both in circulation and in a live preparation, form rouleaux in which several red blood cells stack together similar to coins. While in normal plasma, the rouleaux formations are very short, they are markedly enhanced by the presence of high concentration of certain macromolecules, such as fibrinogen, IgM, igA, igG, dextran, and certain other long macromolecules. Rouleaux formations increase the red cell sedimentation rate, and this rate is often used as an indirect measure of fibrinogen, IgM, and other inducing agents.
- Erythrocyte aggregation is caused by abnormal protein levels as described above in rouleaux formation. Similarly, therapy would be directed at reversing or controlling the particular disease entity. Niacin and HC1 are used in the treatment of red cell aggregation because of their ability to separate red blood cells.
- thrombocyte (platelet) aggregation has been implicated in various heart conditions and strokes.
- Barnett, H.J.M., et al. (1980) Randomized trial of therapy platelet anti-aggregants for threatened strokes. Observations on the pathogeneis, and natural history of threatened stroke, CMA Journal 112, 535; Kalendovsky, Zdenka, et al. (1975), Increased platelet aggreability in young patients with stroke. Archives of Neurology 32,12; Valzhakis, Nicolas, et al.
- OMP Platelet aggregation Adult-onset diabetes mellitus and coronary artery disease, JAMA 239, 732.
- thrombocyte aggregation can be easily discerned in the live cell preparation.
- a variety of natural substances have been found to counteract platelet aggregation, for example, vitamin B6, vitamin C, cod liver oil, vitamin E, garlic, onion, and linseed oil, as well as certain minerals such as magnesium and zinc.
- Treatment for platelet aggregation in the present invention would involve an individualized diet, vitamin, and mineral regimen,in accordance with the above discussion.
- Spicules Spicule formations appear as fibrous-type formations and are frequently associated with rouleaux formation or red cell aggregation, and generally consist of fibrin or alpha-2 macroglobulin, which are the heavier proteins in the blood. Spicule formation, and red cell aggregation, frequently occur with chronic degenerative diseases, and are clinically indicative of a weak liver, as with drug, or alcohol usage. The causative fact or factors for the formation of spicules should be sought. Appropriate treatment according to the present invention includes the use of hydrochloric acid and niacin. 5.
- Chylous Material The presence of chylous material in the live cell preparation, as indicated by the arrow in Figure 5, indicates an abnormally high level of blood fat. Since chylous material can be seen in the live blood cell preparation following meals that are high in fat content, this test is of importance only after a low-fat meal has been consumed. Assuming the chylous material is observed after a small low-fat meal, the test indicates that the patient has high blood fats and/or or a weak liver, that is, disability of the liver to clear fats.
- Treatment would be geared toward fat emulsification using substances such as lecithin, lipotrophic factors such as choline, inositol, and methionine, and instituting a modified diet which include foods low in saturated fats, such as fish and vegetables, and foods high in certain minerals such as iron, known to be helpful for liver function.
- substances such as lecithin, lipotrophic factors such as choline, inositol, and methionine
- a modified diet which include foods low in saturated fats, such as fish and vegetables, and foods high in certain minerals such as iron, known to be helpful for liver function.
- L-Forms are embryonic bacteria that are indicative of infection when observed in the live blood cell preparation. As can be seen in Figure 6, L-forms are similar to the fat particles or chylous material as discussed above, but are generally larger. Patients having L-forms in their blood, as well as other infections discussed below, often have other characteristic conditions such as feeling low and frequently contracting
- OMPI flus colds, or sore throats.
- antibiotics may be used to treat the infection. If the use of antibiotics is not clinically indicated, treatment in the present invention involves the use of nutritional immune stimulators such as vitamins A, C, E, and B6, and other substances such as zinc, pantothenic acid, and thymus.
- nutritional immune stimulators such as vitamins A, C, E, and B6, and other substances such as zinc, pantothenic acid, and thymus.
- Rod Forms are another type of infection that can be seen in the live blood cell preparation. As shown by the arrow in Figure 7, these bacteria appear almost exclusively outside the red blood cells. As indicated above with the L-forms, the type of treatment often depends on the quantity of rod forms that are observed.
- Parasitized Red Blood Cells Another type of infection that can be seen in the live cell preparation are red blood cells that are infected usually by embryonic bacteria but also occassionally by rods or cocci ( Figure 8). Similar accompanying symptoms are observed with parasitized red blood cells as seen with the other types of infection as discussed above. Also, the treatment of parasitized red blood cells is the same as that described in the L-forms and rod forms above and involve the use of antibiotics if the infection is severe or the use of nutritional immune stimulators with a less severe level of infection.
- Protoplast Protoplasts are believed by Dr. V. Livingston (Microbiology of Cancer, Compendium c, 1977) to be large forms of the pleomorphic "cryptocydes" microbe, and classified by Dr. Livingston under the family actinomycetales and have been found to stain intermittently acid fast.
- the appearance of protoplasts indicate multiple degenerative diseases such as scleroderma cancer or rheumatoid arthritis.
- Treatment in part includes the use of immunologic enhancing nutrients such as vitamins A, C, E, and B-6, and zinc, pantothenic acid, thymus, dimethylglycine, and betacarotene. Vaccines and more aggressive nutritional parenteral therapy may be indicated.
- Plaque Plaque formations are similar to protoplasts, the later being more transluscent. Plaque formation occurs coincidentally with heart disease and arteriosclerosis and correlates well with poor circulation. Treatment includes low saturated fat diet and supplements such as vitamin E, niacin, and chondroitin sulfate. 11. Red Crystals
- Figure 11 and identified by characteristic red color (not shown), correlates with toxicity.
- Red crystals are frequently observed in the live cell preparation from patients involved in heavy drug use (as in the photomicrograph in Figure 11) and are also seen in patients with terminal cancer. These patients will frequently- also have a positive bowel toxemia test (urine indican) .
- the treatment of the present invention is to remove the toxic crystals and thereby eliminate the toxicity accompanying the crystal formation with digestive enzymes, acidophilous or yogurt, coupled with a conventional bowel cleanser. Vaccines and more aggressive nutritional parenteral therapy may be indicated. 12.
- Acanthocytes are red blood cells which have lost their discoid shape and which have formed on their surface spicules or burrs of uneven length irregularly distributed over the entire surface. As can be seen in Figure 12, in addition to the burr-like extensions on the acanthocyte, acanthocytes appear smaller than the normal red blood cells having assumed a more spheroidal shape.
- acanthocytes in excess of 1 percent is a valuable diagnostic sign and can indicate certain pathological conditions such as abetalipoproteinemia, disorders of lipid metabolism, alcoholic cirrhosis, and in rare cases of neonatal or other acquired hepatitis. Further, the more deformation of the red blood cell surface, the worse the oxygen exchange at the cellular level. Therefore, if more than 1 percent of these types of red cells are observed in the live cell preparation, the reason for their occurrence should be sought. Treatment includes the use of antioxidants and a low saturated fat diet. 13. Neutrophilic Viability
- a characteristic neutrophil as shown in Figure 13, usually possesses three distinct nuclear segments or lobes. This cell is the primary defense to pyogenic organisms, phagocytizsing the alien invaders. Neutrophils represent more than one-half of the total leukocytes found in a normal live cell preparation, and the activity, and presumably the phagocytoability of the neutrophil, can be evaluated by observing their numbers in the live cell preparation. Further, as indicated by the research of Johnston, et al. (New England Journal of Medicine , August 12, 1982) oxygen is necessary for the phagocytic activity of the neutrophil. The greater the red cell clumping or aggregation, the more inactive the neutrophils appear; and it is assumed that less oxygen is present. Treatment would include hydrochloric acid, vitamins C and A, zinc and
- OMPI other immune enchancers as well as dietary manipulation.
- Hypersegmentation of Neutrophils The hypersegmentation of neutrophil nuclei as illustrated in Figure 14 wherein the neutrophils have five or more lobes and can be routinely diagnosed as megaloblastic anemia. With rare exceptions, megaloblastic anemia responds completely either to parenteral vitamin B-12 or to folate, these disease correlating well with folic acid deficiency (Bills T. and L. Spatz, Neutrophilic Hypersegmentation and Folate Deficiency, American Journal of Clinical Pathology, Vol. 68, 1977). In particular, the finding that any neutrophil has 5 or more nuclear segments, that more than 5 percent has more than 5 segments, or that the majority possess 4 or more segments, signifies megaloblastic hypersegmentation.
- Eosinophils normally constitute about 5 to 10 percent of the granulocyte population and are normally identical in size to neutrophils. Although they are normally identified in standard size preparations by the
- OMPI brilliant orange or red refractile granules that are peroxidase rich, they may also be identified using the microscopic technique of the present invention by their unique nuclear segmentation.
- the eosinophil being the large cell in the center of the photomicrograph, and comparing this eosinophil in Figure 15 with the neutrophil in Figure 14, it can be seen that the eosinophils generally have one or more typically two large round or potato-shaped lobes; they also appear much brighter than neutrophils.
- the population of eosinophils is increased in allergy and parasitic involvement; also, certain tumors . such as eosinophilic granuloma also cause an increase in the production of eosinophils.
- the cause of the increase in eosinophils should be sought and treated accordingly.
- the present invention utilizes vitamins A, C, E, and B-6, and pantothenic acid as treatment.
- Target cells are red blood cells of broader diameter and diminished thickness, which have an artifactual bull's-eye appearance when viewed with dark field microscopy as in the present invention, and are illustrated by an arrow in Figure 16. Targeting is most uniform and striking in hemoglobin C disease and interhapatic or extrahapatic obstructive jaundice. In such diseases, target cells are obvious and very numerous, constituting over half the red blood cell population. Targeting can also be seen in normal individuals postsplenectomy and in rare individuals having a familial deficiency of lecithin-cholesteral acyltransferase.
- target cells are numerous but intermingled with other aberrant cells; such disorders, include sickle cell anemia, most thalassemic disorders, as well as a feature of iron deficiency (hypochromic) anemia. Once the other aberrant cells are identified, the disorder or anemia is treated using iron supplements.
- Anisocytosis is characterized by extreme variations in cell size as indicated in Figure 17; the majority of red cells are conspicuously macrocytic, although mingled with these in less numbers are cells about half-normal in size as indicated by the arrow in Figure 17. Typically, as the anemia worsens, the diverse cell population gravitates towards the extreme in sizes. Generally, this condition of different cell size is caused by a severe anemia such as hypochromic or megaloblastic anemia and is treated, as indicated above, with vitamin B-12, folate, iron supplements, and special diet. 18.
- OMPI Morphologic heterogeneity that occurs in severly hypochromic red cells is often characterized by variations in shape (poikilocytosis) as well as variations in size
- Bland was designed to determine if the cell wall could be protected from oxidative damage and subsequent deformation by treating the patient with antioxidants. Specificly, he administered 600 units of vitamin E which subsequently prevented the oxidated damage. Therefore, based upon this research, the present invention involves the treatment of the poikilocytic red blood cells using an antioxidant such as vitamin E thereby supplementing the apparent deficiency of normal blood antioxidants.
- ovalocytes have also been associated with iron, folic acid, and B-12 deficiencies.
- macrocytes which are cells that have diameter greater than 9 microns (the average diameter of red blood cell being 7.2 microns) and are indicated by the arrow in Figure 20, is- generally found in B-12 and folic acid deficiencies, myxedema, and in hemolytic anemias. Once the cause of this deformity is determined, based on the appearance of other cells in the live cell preparation, appropriate treatment is prescribed.
- Microcytes are defined by a red blood cell having a diameter of less than 6 microns and frequently characterized with a mean corpuscular volume of less than 80 to 82 cubic microns. Microcytic cells generally have less hemoglobin than normal cells and they are therefore frequently seen in iron deficiency anemia and spherocytic and mediterranean hemolytic anemias. The microcyte typically appears as indicated by the arrow in Figure 21.
- OMPI fungal forms can also be observed in sputum samples, as illustrated by the arrows in Figure 25.
- the present invention describes a method of performing an analysis of a live blood cell preparation to detect and identify pathological conditions that are manifested by pathophysiological alterations of blood cells. Further, the method dispenses with the expense of time-consuming preparation and staining of the conventional blood smear by utilizing dark-field microscopy thereby permitting the attending physician to examine and evaluate the preparation while the patient simultaneously views the same preparation. This serves both as a means of visually instructing the patient in the observed abnormalities and as a means of providing the patient with an immediate feedback and diagnosis of any observed pathological conditions requiring treatment.
- the present invention involves a method of treatment that utilizes natural substances, vitamin and mineral supplements, and modified diets.
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Food Science & Technology (AREA)
- Analytical Chemistry (AREA)
- Urology & Nephrology (AREA)
- Hematology (AREA)
- Biomedical Technology (AREA)
- Medicinal Chemistry (AREA)
- Physics & Mathematics (AREA)
- Molecular Biology (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
A method of performing an analysis of a live blood cell preparation to detect and identify pathological conditions that are manifested by pathophysiological alterations of blood cells. The determination of cellular aberrations and inclusions is determined on a live blood cell preparation that are subjected to dark-field microscopic examination immediately after it has been withdrawn from a patient. The method utilizes a microscope with a video camera attachment thereby permitting simultaneously viewing of the live blood cell preparation by both the attending technician and the patient. Further, the invention involves a method of treatment that utilizes natural substances, vitamin and mineral supplements, and modified diets.
Description
IMPROVED METHOD OF LIVE BLOOD CELL ANALYSIS
BACKGROUND OF THE INVENTION The diagnosis of certain diseases or conditions of the body often depends to a large extent on microscopic examination of blood or bone marrow smears in which certain blood disorders appear that are indicative of a disease or condition. Although recent technical advances have essentially automated cell-counting procedures, the clinical hematology laboratory still depends on the visual inspection of smears under the microscope in order to discern blood disorders.
The gradual development of the techniques that led to the present day hematologic practice began over a century ago. Modern techniques basically involve the production of a blood smear on a slide, the differential staining of the slide, which is then followed by microscopic inspection and evaluation. The preparation of a smear on a slide generally involves the use of a "pre-cleaned" slide upon which a drop of blood or dilutent containing the cells is placed. The .cells are then uniformly spread over the slide generally by the use of a coverslip that is brought in contact with the slide and the cells with the slip being moved rapidly from one end of the slide to the other, thereby spreading the cells by capillary action along the line of contact of the slide
and coverslip. The thickness of the smear is a function of the speed with which the coverslip is moved across the slide. The smearing technique must be such that in the final film, the cells generally lie separated side by side with some cells being aggregated in small rouleaux. Smears that are made too thick usually result in overstaining and some clumping, making microscopic analysis of the cells practically impossible. The acquisition of a proper smearing technique is essential to clinical hematology and can only be acquired by practical experience. The smears must then be dried rapidly in order to preserve the shape of the majority of the cells; drying may be' accomplished by the rapid movement of the slide, through the air, the use of previously heated slides, or the preparation of smears under an infrared lamp. Care must be taken in both the preparation of the smear and the drying in order to minimize the creation of artifacts making the appearance of cells in the smears different from those occuring naturally in the circulation. Once a properly made smear has been made and the smear appropriately dried, the preparation must then be fixed with a material such as methanol to prevent subsequent changes in the cells such as distortion or hemolysis when subjected to humidity or staining solutions. When the smear has been properly fixed, the
OMP WIP
preparation is subjected to certain standard staining procedures. The most widely used staining procedure (Pappenheim) involves the use of May-Grunwald stain followed by a distilled water rinse then subsequently stained with dilute Giemsa solution. Generally, the staining technique must be adapted to the requirements of the individual laboratory based on the pH of the distilled water and the frequency of use and replacement of the staining stock solutions. Another common staining procedure is to use commercially available Wright's Stain. Other special staining and cytochemical (histochemical) methods are also available.
Once the slide has been properly prepared, a routine microscopic examination of the smear specimen is carried out under low objective power to obtain a rapid indication of the quantity and quality of the cells, which is then followed by a microscopic examination using both high dry lens and oil emersion lens to make the necessary cytological diagnosis. Considering the vast number of hematological studies that are performed daily, the development of automated techniques would seem eminent. However, at present, the developments with objectives of automatically reading the cells or entirely eliminating the need for smears, staining and microscopic examination, have met with only limited success. Except for standard
cell counting instruments and instruments that have been proposed for the automatic spreading of blood on slides, basic clinical hematological study still involves tedious preparation of smears and the sensitive time-consuming steps of fixing and staining the slide preparation.
Typically, the preparation of a slide requires several hours, and must be accomplished in a laboratory established for that purpose. Therefore, the microscopic examination and evaluation must necessarily occur sometime after the blood has been withdrawn from the patient. Because of the time and special facilities necessary to prepare the slides and perform the microscopic examination, generally the attending physician does not perform the slide evaluation and, further, the results are generally not available until the following day and then presented in such a manner, typically by telephone, as to be useful only to the attending physician and, therefore, have less effect on the patient who does not have an opportunity to view the blood preparations. Thus, in view of the technical expertise required to prepare blood smears for hematological analysis, the necessity of specialized facilities, technicians, and materials, the fact that the attending physican does not perform the evluation of the blood because of the inconvenience, and, therefore, the impossibility of
providing immediate results, diagnosis and prescribed treatment to the patient at the time the blood is drawn, it is highly desirable to provide a method by which a clinical hematological study can be done by an attending physican in the presence of a patient at the time the blood is drawn especially to enhance compliance.
SUMMARY OF THE INVENTION Accordingly, the present invention provides a method of detecting, diagnosing, and prescribing treatment for a variety of pathological conditions that are generally manifested in hematological abnormalities. In particular, the invention provides a method by which live blood cells can be subjected to microscopic examination immediately after blood has been withdrawn from the patient and dispensing with the expensive and time-consuming necessity of- preparing a standard blood smear slide. In addition, the method utilizes a microscope with a dark field condenser and with a video camera attachment thereby permitting simultaneous viewing of the live blood cell preparation by both the attending physician and the patient. The method thus permits the attending physician or techician to evaluate immediately the blood preparation, and provides an immediate feedback to the patient on the evaluation and diagnosis of the blood, while offering a visual presentation of the cells
OMPI
that the attending physican may use to actually show the patient any abnormalities observed.
BRIEF DESCRIPTION OF THE DRAWINGS Figures 1-23 are photomicrographs of live blood cell preparations as viewed under conventional dark field illumination using a 100X oil emersion objective, except for Figure 3 which utilizes a 40X objective; and
Figures 24 and 25 are photomicrographs of sputum preparations viewed under conventional dark field illumination, utilizing a 100X oil emersion objective. DETAILED DESCRIPTION OF THE INVENTION The basis of the present invention described is the detection and identification of pathophysiologically altered cells in a live state. Although these morphological changes and inclusions in cell preparations can be visualized using standard blood smear preparations, the advent of phase contrast, dark field, fluorescent, electron, and scanning electron microscopy have made the identification of many morphological features of cells possible without the need for the more complicated and difficult slide preparation and staining procedures.
In particular, the invention uses dark field microscopy which profoundly enhances morphological characteristics of living cells such as size, shape, and type of cells, as well as major inclusions within the cell
and abnormal constituents of the surrounding fluid. The invention has compiled a series of the more common pathophysiological states of living cells that can be identified by morphological changes or abnormal inclusions and has reinterpreted these findings to diagnose a pathological state or condition of the body manifested by the pathophysiological state of the living cells. More importantly, the evaluation of the live cell preparation is done within minutes after having withdrawn the blood from the patient and is accomplished in the presence of the patient such that the attending physician or technician may point out the differences between the normal state and any observed pathophysiological state of the cells. Therefore, the invention provides a blood examination procedure that may also be used by the attending physician to clearly illustrate to the patient abnormalities that may occur in the blood. The patient not only receives diagnostic information that can be readily understood, but the entire procedure of drawing the patient's blood, evaluation of the blood preparation, diagnosis, explanation of the diagnostic results to the patient, and the prescribing of a treatment for any abnormal conditions that may be found, may be accomplished within a short period of time such as less than one hour. The instrumentation used in the microscopic
examination of the living cell preparation includes a standard bright field microscope (Bausch and Lomb, Model
DRT3Z1H) that has been equipped with 10X, 20X, 40X, and
100X (oil) planachromatic objectives, and a turret-type condenser which includes dark field capability. The microscope is additionally configured with a conventional video camera that is mounted on the microscope, the image of which-is displayed on a standard television set.
The procedure involved for obtaining a blood specimen and preparing the live blood cell preparation . involves obtaining a sample of blood from a patient using a standard finger prick. In particular, the patient's finger is cleaned well with an alcohol swab then wiped dry with lint-free tissue. Once the finger has been pricked with a lancet and a drop of blood has formed on the finger, the center of a precleaned glass slide is touched gently to the drop of blood taking care not to touch any portion of the finger. The slide is subsequently rotated such that the blood sample is now positioned on the top surface of the slide and a standard coverslip (22x40) is then placed on the slide over the sample of blood. Following standard microscopy techniques, a drop of emersion oil is placed between the slide and the condenser and a 10X objective is used to scan the blood preparation to ascertain that portion of the blood preparation where
the blood appears the thinnest and the cells most evenly distributed. A drop of oil is then placed on the coverslip and the 100X oil emersion objective is used for the evaluation of the blood cell preparation, except when thrombocyte aggregations are to be viewed, wherein the 40X objection is occasionally used (without oil). Of course, those skilled in the art will recognize the procedure described above refers to the preparation of a blood sample and that the procedure can be used for the preparation of other samples such as sputum and urine. For the purpose of this application, the term blood sample shall be defined to include blood, sputum, urine, and other body fluid specimens.
The live blood cell preparation is then searched for abnormalities, each of which is separately discussed below:
1. Rouleaux Formation
In the circulation, mature red blood cells
(erythrocytes) have a general biconcave shape. Normally, these red blood cells both in circulation and in a live preparation, form rouleaux in which several red blood cells stack together similar to coins. While in normal plasma, the rouleaux formations are very short, they are markedly enhanced by the presence of high concentration of
certain macromolecules, such as fibrinogen, IgM, igA, igG, dextran, and certain other long macromolecules. Rouleaux formations increase the red cell sedimentation rate, and this rate is often used as an indirect measure of fibrinogen, IgM, and other inducing agents. The finding of excessive rouleaux in the live preparation (Figure 1) is indicative that certain macroglobulin levels are high and that the most common causative agent should be sought depending on the excessivity of the rouleaux formation. Since red cells normally must squeeze through capillaries in single file in order to exchange oxygen for carbon dioxide, rouleaux formations would cause less oxygen to be transferred to the cell, therefore, resulting in fatigue. Conventional anti-coagulant therapy has been found helpful in reducing the degree of rouleaux formations. Hydrochloric acid (HC1) and niacin have been shown clinically to separate rouleaux formation, and these natural substances are used for the treatment of rouleaux formations in the present invention.
2. Erythrocyte Aggregation Erythrocyte aggregation, as can be seen in Figure
2, is similar to rouleaux formation except the condition is usually worse and rouleaux formation is often seen ,as clumps of red cells. This condition is obviously worse than rouleaux formation and has a more negative influence
-li¬ on oxygen exchange.
Erythrocyte aggregation is caused by abnormal protein levels as described above in rouleaux formation. Similarly, therapy would be directed at reversing or controlling the particular disease entity. Niacin and HC1 are used in the treatment of red cell aggregation because of their ability to separate red blood cells.
3. Thrombocyte Aggregation Recently, thrombocyte (platelet) aggregation has been implicated in various heart conditions and strokes. For example, Barnett, H.J.M., et al. (1980), Randomized trial of therapy platelet anti-aggregants for threatened strokes. Observations on the pathogeneis, and natural history of threatened stroke, CMA Journal 112, 535; Kalendovsky, Zdenka, et al. (1975), Increased platelet aggreability in young patients with stroke. Archives of Neurology 32,12; Valzhakis, Nicolas, et al. (1979), Platelet aggregation in relationship to plasma catecholamines in patients With hypertension, Atherosclerosis 32,451; Mehta, Jawahar, et al. (1981), Platelet function in hypertension and effective therapy, American Journal of Cardiology, 47,331; Al-Mefty, Osama, et al. (1979), Transient ischemic attacks due to increased platelet aggregation and adhesiveness. Journal of Neurology 50,449; and Davis, James W., et al. (1978),
OMP
Platelet aggregation, Adult-onset diabetes mellitus and coronary artery disease, JAMA 239, 732. Referring to Figure 3, thrombocyte aggregation can be easily discerned in the live cell preparation. A variety of natural substances have been found to counteract platelet aggregation, for example, vitamin B6, vitamin C, cod liver oil, vitamin E, garlic, onion, and linseed oil, as well as certain minerals such as magnesium and zinc. Treatment for platelet aggregation in the present invention would involve an individualized diet, vitamin, and mineral regimen,in accordance with the above discussion.
4. Spicules Spicule formations, as can be seen in Figure 4, appear as fibrous-type formations and are frequently associated with rouleaux formation or red cell aggregation, and generally consist of fibrin or alpha-2 macroglobulin, which are the heavier proteins in the blood. Spicule formation, and red cell aggregation, frequently occur with chronic degenerative diseases, and are clinically indicative of a weak liver, as with drug, or alcohol usage. The causative fact or factors for the formation of spicules should be sought. Appropriate treatment according to the present invention includes the use of hydrochloric acid and niacin. 5. Chylous Material
The presence of chylous material in the live cell preparation, as indicated by the arrow in Figure 5, indicates an abnormally high level of blood fat. Since chylous material can be seen in the live blood cell preparation following meals that are high in fat content, this test is of importance only after a low-fat meal has been consumed. Assuming the chylous material is observed after a small low-fat meal, the test indicates that the patient has high blood fats and/or or a weak liver, that is, disability of the liver to clear fats. Treatment would be geared toward fat emulsification using substances such as lecithin, lipotrophic factors such as choline, inositol, and methionine, and instituting a modified diet which include foods low in saturated fats, such as fish and vegetables, and foods high in certain minerals such as iron, known to be helpful for liver function.
6. L-Forms L-Forms are embryonic bacteria that are indicative of infection when observed in the live blood cell preparation. As can be seen in Figure 6, L-forms are similar to the fat particles or chylous material as discussed above, but are generally larger. Patients having L-forms in their blood, as well as other infections discussed below, often have other characteristic conditions such as feeling low and frequently contracting
OMPI
flus, colds, or sore throats.
If the infection is severe, as indicated by the quantity of bacteria observed, antibiotics may be used to treat the infection. If the use of antibiotics is not clinically indicated, treatment in the present invention involves the use of nutritional immune stimulators such as vitamins A, C, E, and B6, and other substances such as zinc, pantothenic acid, and thymus.
7. Rod Forms Rod forms are another type of infection that can be seen in the live blood cell preparation. As shown by the arrow in Figure 7, these bacteria appear almost exclusively outside the red blood cells. As indicated above with the L-forms, the type of treatment often depends on the quantity of rod forms that are observed.
8. Parasitized Red Blood Cells Another type of infection that can be seen in the live cell preparation are red blood cells that are infected usually by embryonic bacteria but also occassionally by rods or cocci (Figure 8). Similar accompanying symptoms are observed with parasitized red blood cells as seen with the other types of infection as discussed above. Also, the treatment of parasitized red blood cells is the same as that described in the L-forms and rod forms above and involve the use of antibiotics if
the infection is severe or the use of nutritional immune stimulators with a less severe level of infection.
9. Protoplast Protoplasts, as shown in Figure 9, are believed by Dr. V. Livingston (Microbiology of Cancer, Compendium c, 1977) to be large forms of the pleomorphic "cryptocydes" microbe, and classified by Dr. Livingston under the family actinomycetales and have been found to stain intermittently acid fast. The appearance of protoplasts indicate multiple degenerative diseases such as scleroderma cancer or rheumatoid arthritis. Treatment in part includes the use of immunologic enhancing nutrients such as vitamins A, C, E, and B-6, and zinc, pantothenic acid, thymus, dimethylglycine, and betacarotene. Vaccines and more aggressive nutritional parenteral therapy may be indicated.
10. Plaque Plaque formations, as illustrated in Figure 10, are similar to protoplasts, the later being more transluscent. Plaque formation occurs coincidentally with heart disease and arteriosclerosis and correlates well with poor circulation. Treatment includes low saturated fat diet and supplements such as vitamin E, niacin, and chondroitin sulfate. 11. Red Crystals
OMPI
The formation of red crystals, as illustrated in
Figure 11 and identified by characteristic red color (not shown), correlates with toxicity. Red crystals are frequently observed in the live cell preparation from patients involved in heavy drug use (as in the photomicrograph in Figure 11) and are also seen in patients with terminal cancer. These patients will frequently- also have a positive bowel toxemia test (urine indican) . The treatment of the present invention is to remove the toxic crystals and thereby eliminate the toxicity accompanying the crystal formation with digestive enzymes, acidophilous or yogurt, coupled with a conventional bowel cleanser. Vaccines and more aggressive nutritional parenteral therapy may be indicated. 12. Acanthocyte Percentage
Acanthocytes are red blood cells which have lost their discoid shape and which have formed on their surface spicules or burrs of uneven length irregularly distributed over the entire surface. As can be seen in Figure 12, in addition to the burr-like extensions on the acanthocyte, acanthocytes appear smaller than the normal red blood cells having assumed a more spheroidal shape.
The presence of acanthocytes in excess of 1 percent is a valuable diagnostic sign and can indicate certain pathological conditions such as
abetalipoproteinemia, disorders of lipid metabolism, alcoholic cirrhosis, and in rare cases of neonatal or other acquired hepatitis. Further, the more deformation of the red blood cell surface, the worse the oxygen exchange at the cellular level. Therefore, if more than 1 percent of these types of red cells are observed in the live cell preparation, the reason for their occurrence should be sought. Treatment includes the use of antioxidants and a low saturated fat diet. 13. Neutrophilic Viability
A characteristic neutrophil, as shown in Figure 13, usually possesses three distinct nuclear segments or lobes. This cell is the primary defense to pyogenic organisms, phagocytizsing the alien invaders. Neutrophils represent more than one-half of the total leukocytes found in a normal live cell preparation, and the activity, and presumably the phagocytoability of the neutrophil, can be evaluated by observing their numbers in the live cell preparation. Further, as indicated by the research of Johnston, et al. (New England Journal of Medicine , August 12, 1982) oxygen is necessary for the phagocytic activity of the neutrophil. The greater the red cell clumping or aggregation, the more inactive the neutrophils appear; and it is assumed that less oxygen is present. Treatment would include hydrochloric acid, vitamins C and A, zinc and
OMPI
other immune enchancers as well as dietary manipulation. 14. Hypersegmentation of Neutrophils The hypersegmentation of neutrophil nuclei as illustrated in Figure 14 wherein the neutrophils have five or more lobes and can be routinely diagnosed as megaloblastic anemia. With rare exceptions, megaloblastic anemia responds completely either to parenteral vitamin B-12 or to folate, these disease correlating well with folic acid deficiency (Bills T. and L. Spatz, Neutrophilic Hypersegmentation and Folate Deficiency, American Journal of Clinical Pathology, Vol. 68, 1977). In particular, the finding that any neutrophil has 5 or more nuclear segments, that more than 5 percent has more than 5 segments, or that the majority possess 4 or more segments, signifies megaloblastic hypersegmentation.
In addition, as illustrated by Figure 14 wherein the hypersegmented neutrophil appears as the large white cell in the center of the photomicrogragh, the appearance of ovalocytes (immediately right of the neutrophil) also commonly occurs with megaloblastic anemia.
15. Eosinophils
Eosinophils normally constitute about 5 to 10 percent of the granulocyte population and are normally identical in size to neutrophils. Although they are normally identified in standard size preparations by the
OMPI
brilliant orange or red refractile granules that are peroxidase rich, they may also be identified using the microscopic technique of the present invention by their unique nuclear segmentation. As illustrated in Figure 15, the eosinophil being the large cell in the center of the photomicrograph, and comparing this eosinophil in Figure 15 with the neutrophil in Figure 14, it can be seen that the eosinophils generally have one or more typically two large round or potato-shaped lobes; they also appear much brighter than neutrophils.
The population of eosinophils is increased in allergy and parasitic involvement; also, certain tumors . such as eosinophilic granuloma also cause an increase in the production of eosinophils. The cause of the increase in eosinophils should be sought and treated accordingly. The present invention utilizes vitamins A, C, E, and B-6, and pantothenic acid as treatment.
16. Target Cells Target cells are red blood cells of broader diameter and diminished thickness, which have an artifactual bull's-eye appearance when viewed with dark field microscopy as in the present invention, and are illustrated by an arrow in Figure 16. Targeting is most uniform and striking in hemoglobin C disease and interhapatic or extrahapatic obstructive jaundice. In such
diseases, target cells are obvious and very numerous, constituting over half the red blood cell population. Targeting can also be seen in normal individuals postsplenectomy and in rare individuals having a familial deficiency of lecithin-cholesteral acyltransferase. Further, in many other disorders, target cells are numerous but intermingled with other aberrant cells; such disorders, include sickle cell anemia, most thalassemic disorders, as well as a feature of iron deficiency (hypochromic) anemia. Once the other aberrant cells are identified, the disorder or anemia is treated using iron supplements.
17. Anisocytosis Anisocytosis is characterized by extreme variations in cell size as indicated in Figure 17; the majority of red cells are conspicuously macrocytic, although mingled with these in less numbers are cells about half-normal in size as indicated by the arrow in Figure 17. Typically, as the anemia worsens, the diverse cell population gravitates towards the extreme in sizes. Generally, this condition of different cell size is caused by a severe anemia such as hypochromic or megaloblastic anemia and is treated, as indicated above, with vitamin B-12, folate, iron supplements, and special diet. 18. Poikilocytosis
OMPI
Morphologic heterogeneity that occurs in severly hypochromic red cells is often characterized by variations in shape (poikilocytosis) as well as variations in size
(anisocytosis), as indicated as above. These variations in the shape of red blood cells is illustrated in Figure 18 as indicated by the arrows. Recent work, however, by Bland, et al. (Effect of alpha-tocopherol on the rate of photohemolysis of human erythrocytes, Physiol. Chem. Physics 7:69, 1975) and Horwitt, et al. (Effects of limited tocopherol intake in man with relationship to erythrocyte hemolysis, Am. J. Clin. Nutr. 8:451, 1960) has shown that these cells had an increased tendency to hemolyze, since these cell membranes containing a quantity of unsaturated fat and other fats. The research of Dr. Bland was designed to determine if the cell wall could be protected from oxidative damage and subsequent deformation by treating the patient with antioxidants. Specificly, he administered 600 units of vitamin E which subsequently prevented the oxidated damage. Therefore, based upon this research, the present invention involves the treatment of the poikilocytic red blood cells using an antioxidant such as vitamin E thereby supplementing the apparent deficiency of normal blood antioxidants.
19. Ovalocytes As indicated above in test number 14,
OMPI
hypersegmentation of neutrophils, the appearance of ovalocytes has also been associated with iron, folic acid, and B-12 deficiencies. A population of 15 to 25 percent ovalocytes in a live cell preparation, as indicated by the arrows in Figure 19, strongly indicates the presence of hypochromic microcytic anemia, and is generally treated as described above in hypersegmentation of neutrophils.
20. Macrocytes
The occurrence of macrocytes, which are cells that have diameter greater than 9 microns (the average diameter of red blood cell being 7.2 microns) and are indicated by the arrow in Figure 20, is- generally found in B-12 and folic acid deficiencies, myxedema, and in hemolytic anemias. Once the cause of this deformity is determined, based on the appearance of other cells in the live cell preparation, appropriate treatment is prescribed.
21. Microcytes
Microcytes are defined by a red blood cell having a diameter of less than 6 microns and frequently characterized with a mean corpuscular volume of less than 80 to 82 cubic microns. Microcytic cells generally have less hemoglobin than normal cells and they are therefore frequently seen in iron deficiency anemia and spherocytic and mediterranean hemolytic anemias. The microcyte
typically appears as indicated by the arrow in Figure 21. Once the cause of the production of microcytes is determined based on the appearance of other aberrant cells in the live cell preparation as discussed above in the macrocyte test, appropriate treatment is prescribed.
22. Fungal Forms In Blood In addition to the detection and identification of pathophysiological conditions of blood, various tests of the present invention also involve the detection and identification of other forms of microorganisms both in. the blood and in sputum. Referring to Figure 22 as indicated by the arrow, there is shown a fungal organism in the live cell preparation. Natural immune stimulators such as biotin, zinc, and vitamins A and C are prescribed for treatment.
23-25. Sputum Preparation The method described in the invention for the preparation of blood for live cell analysis can be easily adapted to analyze samples of sputum for the presence of certain types of pathogenic microorganisms. For example, as shown in Figure 23, there is shown a fusiform of bacteria observed in a sample of sputum taken from the mouth of a patient with the Vincent's infection (trench mouth), gingiva-stomatitis. Treponema was also observed in the same preparation (Figure 24). Various types of other
OMPI
fungal forms can also be observed in sputum samples, as illustrated by the arrows in Figure 25.
In summary, the present invention describes a method of performing an analysis of a live blood cell preparation to detect and identify pathological conditions that are manifested by pathophysiological alterations of blood cells. Further, the method dispenses with the expense of time-consuming preparation and staining of the conventional blood smear by utilizing dark-field microscopy thereby permitting the attending physician to examine and evaluate the preparation while the patient simultaneously views the same preparation. This serves both as a means of visually instructing the patient in the observed abnormalities and as a means of providing the patient with an immediate feedback and diagnosis of any observed pathological conditions requiring treatment. In addition, the present invention involves a method of treatment that utilizes natural substances, vitamin and mineral supplements, and modified diets.
Claims
What is claimed is: 1. The method of detecting and treating pathological conditions comprising the steps of: obtaining a blood sample from a patient; applying the sample to a microscope slide; covering the sample on the slide with a coverslip; viewing the sample on the microscope slide through a microscope to view the sample; displaying the microscope field so as to be directly viewed simultaneously by a technician and a patient; identifying blood cell aberations in the sample; informing the patient while viewing the displayed sample of any observed blood cell aberations; and prescribing diet supplements in response to the identification of predetermined cell aberations.
2. The method of claim 1 wherein the simultaneous viewing of the microscope field comprises displaying the microscope field on a video screen.
3. The method of claim 1 wherein the viewing of the microscope slide comprises the use of dark field
OMPI S IPO microscopy .
4. The method of claim 1 wherein said identification step comprises the observation of rouleaux formation of more than three red blood cells per formation.
5. The method of claim 1 wherein said identification step comprises the observation of erythrocyte aggregation.
6. The method of claim 1 wherein said identification step comprises the observation of thrombocyte aggregation.
7. The method of claim 1 wherein said identification step comprises the observation of spicule formations.
8. The method of claim 1 wherein said identification step comprises the observation of chylous material.
9. The method of claim 1 wherein said identification step comprises the observation of L-forms.
10. The method of claim 1 wherein said identification step comprises the observation of rod forms.
11. The method of claim 1 wherein said identification step comprises the observation of parasitized red blood cells.
12. The method of claim 1 wherein said identification step comprises the observation of protoplasts.
13. The method of claim 1 wherein said identification step comprises the observation of plaque.
14. The method of claim 1 wherein said identification step comprises the observation of red crystals.-
15. The method of claim 1 wherein said identification step comprises the observation of acanthocyte percentage in excess of 1 percent of the red blood cells.
16. The method of claim 1 wherein said identification step comprises the observation of the activity of neutrophils.
17. The method of claim 1 wherein said identification step comprises the observation of hypersegmentation of neutrophils, further comprising the observation of any neutrophil having more than five nuclear segments, more than 5 percent having five segments or more, or a majority of neutrophils possessing four or more segments.
18. The method of claim 1 wherein said identification step comprises the observation of eosinophils in excess of 5 - 10 percent of the granulocyte
OMPI population.
19. The method of claim 1 wherein said identification step comprises the observation of target cells.
20. The method of claim 1 wherein said identification step comprises the observation of anisocytes*.
21. The method of claim 1 wherein said identification step comprises the observation of poikilocytes.
22. The method of claim 1 wherein said identification step comprises the observation of ovalocytes.
23. The method of claim 1 wherein said identification step comprises the observation of macrocytes.
24. The method of claim 1 wherein said identification step comprises the observation of microcytes.
25. The method of claim 1 wherein said identification step comprises the observation of microorganisms.
26. The method of claim 4 wherein said prescribing step comprises prescribing hydrochloric acid and niacin.
27. The method of claim 5 wherein said prescribing step comprises prescribing hydrochloric acid and niacin.
28. The method of claim 6 wherein said prescribing step comprises prescribing vitamins B-6, C, E and cod liver oil.
29. The method of claim 7 wherein said prescribing step comprises prescribing hydrochloric acid and niacin.
30. The method of claim 8 wherein said prescribing step comprises prescribing lecithin and low ' saturated fat diet.
31. The method of claim 9 wherein said prescribing step comprises prescribing vitamins A, C, E, and B-6.
32. The method of claim 10 wherein said prescribing step comprises prescribing vitamins A, C, E, and B-6.
33. The method of claim 11 wherein said prescribing step comprises prescribing vitamin A, C, E, and B-6.
34. The method of claim 12 wherein said prescribing step comprises prescribing vitamins A, C, E, and B-6, and zinc, pantothenic acid, thymus, dimethylglycine, and betacarotene. ^^TREX
OMPI
V- WIPO
35. The method of claim 13 wherein said prescribing step comprises prescribing low saturated fat diets and diet supplements vitamin E, niacin, and chondroitin sulfate.
36. The method of claim 14 wherein said prescribing step comprises prescribing acidophilous and conventional bowel cleansers.
37. The method of claim 15 wherein said prescribing step comprises prescribing antioxidants and a low saturated fat diet cleansers and lactobacillus.
38. The method of claim 16 wherein said prescribing step comprises prescribing vitamins C and A, zinc, and a special diet.
39. The method of claim 17 wherein said prescribing step comprises prescribing vitamin B-12 and folate.
40. The method of claim 18 wherein said prescribing step comprises prescribing vitamins A, C, E, and B-6, and pantothenic acid.
41. The method of claim 19 wherein said prescribing step comprises prescribing iron supplements.
42. The method of claim 20 wherein said prescribing step comprises prescribing vitamin B-12, folate, iron supplements, and a specialized diet.
43. The method of claim 21 wherein said prescribing step comprises prescribing vitamin E.
44. The method of claim 22 wherein said prescribing step comprises prescribing vitamin B-12 and folate.
45. The method of claim 23 wherein said prescribing step comprises prescribing folic acid and
B-12.
46. The method of claim 24 wherein said prescribing step comprises prescribing iron supplements.
47. The method of claim 25 wherein said prescribing step comprises prescribing vitamins A and C, zinc and biotin.
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US54726083A | 1983-10-31 | 1983-10-31 | |
US547,260 | 1990-07-02 |
Publications (1)
Publication Number | Publication Date |
---|---|
WO1985001960A1 true WO1985001960A1 (en) | 1985-05-09 |
Family
ID=24183975
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
PCT/US1984/001673 WO1985001960A1 (en) | 1983-10-31 | 1984-10-17 | Improved method of live blood cell analysis |
Country Status (3)
Country | Link |
---|---|
EP (1) | EP0160038A1 (en) |
AU (1) | AU3550684A (en) |
WO (1) | WO1985001960A1 (en) |
Cited By (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
DE102009059274A1 (en) * | 2009-12-22 | 2011-06-30 | Kraus, Max-Joseph, Dr. med., 82031 | Method for quantifying shape of thrombocytes, involves producing images of thrombocytes, and determining measure for parameters of thrombocytes and/or measure for pseudopodium of thrombocytes based on images |
RU2715552C1 (en) * | 2018-10-18 | 2020-03-02 | Ольга Олеговна Анисимова | Method for digital microscopy of native blood |
Families Citing this family (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
GR850042B (en) * | 1984-01-12 | 1985-04-10 | Kovacs Adam |
Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US3297873A (en) * | 1963-12-30 | 1967-01-10 | Avco Corp | Microscope system with means for viewing the entire image and means for measuring radiation from a selected segment of image |
US3525803A (en) * | 1966-08-01 | 1970-08-25 | Int Research & Dev Co Ltd | Means for detecting malignant cells in human and animal tissue |
-
1984
- 1984-10-17 WO PCT/US1984/001673 patent/WO1985001960A1/en unknown
- 1984-10-17 EP EP84903873A patent/EP0160038A1/en not_active Withdrawn
- 1984-10-17 AU AU35506/84A patent/AU3550684A/en not_active Abandoned
Patent Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US3297873A (en) * | 1963-12-30 | 1967-01-10 | Avco Corp | Microscope system with means for viewing the entire image and means for measuring radiation from a selected segment of image |
US3525803A (en) * | 1966-08-01 | 1970-08-25 | Int Research & Dev Co Ltd | Means for detecting malignant cells in human and animal tissue |
Non-Patent Citations (3)
Title |
---|
Dietetic Foods, issued 1968, A.E. BENDER, New York, Chemical Publishing, page 197 * |
Gradwohl's Clinical Laboratory Methods and Diagnosis, Vols. I and II, issued 1970, SAM FRANKEL et al., Editors, Saint Louis, C.V. MOSBY Company, pp. 8,9,476,477,484-486,501-503,519-532,533-552,1024,1379,1711 * |
The Complete Book of Vitamins, issued 1977, CHARLES GERRAS et al. Editors, Emmaus Pennsylvania, Rodale Press, pp. 216-217,282-287,466-470,471-472 * |
Cited By (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
DE102009059274A1 (en) * | 2009-12-22 | 2011-06-30 | Kraus, Max-Joseph, Dr. med., 82031 | Method for quantifying shape of thrombocytes, involves producing images of thrombocytes, and determining measure for parameters of thrombocytes and/or measure for pseudopodium of thrombocytes based on images |
DE102009059274B4 (en) * | 2009-12-22 | 2012-07-19 | Max-Joseph Kraus | Method for measuring the dynamics of changes in platelets |
RU2715552C1 (en) * | 2018-10-18 | 2020-03-02 | Ольга Олеговна Анисимова | Method for digital microscopy of native blood |
Also Published As
Publication number | Publication date |
---|---|
EP0160038A1 (en) | 1985-11-06 |
AU3550684A (en) | 1985-05-22 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
Lamchiagdhase et al. | Urine sediment examination: a comparison between the manual method and the iQ200 automated urine microscopy analyzer | |
US4393466A (en) | Method of analyzing particles in a dilute fluid sample | |
Cooke et al. | The Polynuclear Count: The Nucleus of the Neutrophil Polymorphonuclear Leucocyte in Health and Disease, with Some Observations on the Macropolycyte | |
EP0179107B1 (en) | Metachromatic dye sorption means for differential determination of sub-populations of lymphocytes | |
JPH03266999A (en) | Reagent for automatic flow sightmetry measurement of sub-population of at least one leukocyte from whole blood and measuring method using said reagent | |
GB2095402A (en) | Metachromatic dye sorption means for differential determination of developmental stages of neutrophilic granulocytic cells and other leukocytes | |
Freemont et al. | Atlas of synovial fluid cytopathology | |
Vander et al. | Reticulocyte counts by means of fluorescence microscopy | |
JPH0473104B2 (en) | ||
Overstreet et al. | Simultaneous assessment of human sperm motility and morphology by videomicrography | |
EP0912891A1 (en) | Method of carrying out blood tests | |
WO1985001960A1 (en) | Improved method of live blood cell analysis | |
Gruner | The Biology of the Blood-Cells: With a Glossary of Hæmatological Terms | |
JP2000502797A (en) | Methods for testing cell samples | |
Maedel et al. | Examination of the peripheral blood film and correlation with the complete blood count | |
Montoya-Navarrete et al. | Red blood cells morphology and morphometry in adult, senior, and geriatricians dogs by optical and scanning electron microscopy | |
Schreinemachers et al. | Effect of residual splenic function and folate levels on the frequency of micronucleated red blood cells in splenectomized humans | |
Miswan et al. | An overview: Segmentation method for blood cell disorders | |
Logan et al. | New spiral bacterium in the gastric mucosa: Gastrospirillum hominis. | |
Alexandratou et al. | Morphometric characteristics of red blood cells as diagnostic factors for coronary artery disease | |
EP0172841B1 (en) | Method for detecting malign proliferation | |
Butina | Body fluid analysis in the hematology laboratory | |
Mukherjee et al. | Application of biomedical image processing in blood cell counting using hough transform | |
Keegan et al. | Fresh capillary blood analysis using darkfield microscopy as a tool for screening nutritional deficiencies of iron and cobalamin (vitamin B12): A validity study | |
Ameri et al. | Analysis of cerebrospinal fluid from clinically healthy Iranian fat-tailed sheep |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
AK | Designated states |
Designated state(s): AU JP |
|
AL | Designated countries for regional patents |
Designated state(s): AT BE CH DE FR GB LU NL SE |