Prognosis and risk assessment in stroke patients by determining the level of marker peptides

Abstract

The present invention relates to a method for prognosis of an outcome or assessing the risk of a patient having suffered a stroke or a transient ischemic attack, comprising the determination of the level of at least one marker peptide in said sample said marker peptide selected from the group comprising ANP, AVP, ADM, ET-1, troponin, CRP, calcitonin and hGH or fragments thereof or its precursor or fragments thereof and attributing the level of said at least one marker peptides its precursor or fragments thereof with the prognosis of an outcome or assessing the risk for said patient.

Claims

1. A method for diagnosing and treating a stroke or a transient ischemic attack in a patient suspected of having a stroke or a transient ischemic attack within 5 days to a year after said patient has previously suffered a stroke or transient ischemic attack, said method comprising: a. obtaining a sample of bodily fluid from said patient, b. detecting and quantitating in said sample from said patient the level of at least one cardiovascular peptide, wherein said cardiovascular peptide is selected from the group consisting of MR-proANP (mid-regional fragment of proANP), CT-proAVP (C-terminal fragment of arginine vasopressin precursor proAVP), MR-proADM (mid-regional fragment of adrenomedullin), CT-proET-1 (C-terminal fragment of endothelin-1 precursor pro-ET-1), PCT (calcitonin precursor procalcitonin), and hGH (human growth hormone), wherein said detection and quantitation comprises contacting the sample with a diagnostic assay reagent comprising a capture probe that specifically binds to said cardiovascular peptide, and detecting and quantitating the thus-formed complexes of said capture probe and cardiovascular peptide, and c. diagnosing said patient as having had a stroke or a transient ischemic attack by comparing the level of the cardiovascular peptide in the patient to a threshold level, wherein a level value of the cardiovascular peptide below the threshold is indicative for transient ischemic attack and a level value of the cardiovascular peptide above the threshold is indicative for stroke, and wherein the threshold level for MR-proANP is from about 90 to about 140 pmol/l, the threshold level for CT-proAVP is from about 9.5 to about 11.5 pmol/l, the threshold level for MR-proADM is from about 0.5 to about 0.8 nmol/l, the threshold level for CT-pro-ET-1 is from about 65 to about 90 pmol/l, the threshold level for PCT is from about 0.0230 to about 0.0260 ng/ml, and the threshold level for hGH is from about 0.10 to about 0.3 ng/ml, and d. treating said patient for a stroke or transient ischemic attack.

2. The method of claim 1, wherein said cardiovascular peptide is CT-proAVP, MR-proADM, or CT-proET-1, and CT-proAVP is amino acids 107-145 of SEQ ID NO:12, MR-proADM is amino acids 45-92 of SEQ ID NO:1, and CT-proET-1 is amino acids 151-195 of SEQ ID NO:17.

3. The method of claim 1, wherein the levels of at least two of said cardiovascular peptides are detected and quantitated.

4. The method of claim 3, wherein the levels of a set of at least two cardiovascular peptides are detected and quantitated and the set of said at least two cardiovascular peptides is selected from the following combinations of cardiovascular peptides: MR-proANP and CT-proAVP, MR-proANP and MR-proADM, MR-proANP and CT-proET-1, MR-proANP and PCT, MR-proANP and hGH, MR-proADM and CT-proET-1, MR-proADM and PCT, MR-proADM and hGH, CT-proET-1 and PCT, CT-proET-1 and hGH, and PCT and hGH.

5. The method of claim 3, wherein the levels of a set of at least two cardiovascular peptides are detected and quantitated and the set of said at least two cardiovascular peptides is selected from the group comprising the following combinations of cardiovascular peptides: MR-proANP and CT-proAVP, MR-proANP and MR-proADM, MR-proANP and CT-proET-1, MR-proANP and PCT, and MR-proANP and hGH.

6. The method of claim 5, wherein the set of said at least two cardiovascular peptides is MR-proANP and MR-proADM.

7. A method for detecting and quantitating the level of at least one cardiovascular peptide in a patient who has suffered a stroke or a transient ischemic attack, comprising: a. obtaining a sample of bodily fluid from said patient, b. detecting and quantitating in said sample from said patient the levels of a set of at least two cardiovascular peptides and the set of said at least two cardiovascular peptides is selected from following combinations of cardiovascular peptides: MR-proANP and CT-proAVP, MR-proANP and MR-proADM, MR-proANP and CT-proET-1, MR-proANP and PCT, MR-proANP and hGH, MR-proADM and CT-proET-1, MR-proADM and PCT, MR-proADM and hGH or CT-proET-1 and PCT, CT-proET-1 and hGH, and PCT and hGH, wherein said detection and quantitation comprises contacting the sample with a diagnostic assay reagent comprising a capture probe that specifically binds to said cardiovascular peptide, and detecting and quantitating the thus-formed complexes of said capture probe and cardiovascular peptide, wherein said detecting and quantitating is conducted on the first day of hospitalization (day 0) of said patient or on day 1 after hospitalization of said patient.

8. The method of claim 7, wherein the levels of a set of at least two cardiovascular peptides are detected and quantitated and the set of said at least two cardiovascular peptides is selected from the group comprising the following combinations of cardiovascular peptides: MR-proANP and CT-proAVP, MR-proANP and MR-proADM, MR-proANP and CT-proET-1, MR-proANP and PCT, and MR-proANP and hGH.

9. The method of claim 8, wherein the set of said at least two cardiovascular peptides is MR-proANP and MR-proADM.

Description

DESCRIPTION OF DRAWINGS

(1) FIG. 1: Kaplan-Meier plot (proportion of patients surviving within four months) for patients with high (dashed lined) and low (solid line) levels of pre-interventionally measured MR-proANP.

(2) FIG. 2: Kaplan-Meier plot for patients with high (dashed lined) and low (solid line) levels of pre-interventionally measured MR-proADM.

(3) FIG. 3: Kaplan-Meier plot for patients with high (dashed lined) and low (solid line) levels of pre-interventionally measured CT-proET-1.

(4) FIG. 4: Kaplan-Meier plot for patients with high (dashed lined) and low (solid line) levels of pre-interventionally measured CT-proAVP.

(5) FIG. 5: Kaplan-Meier plot for patients with high (dashed lined) and low (solid line) levels of pre-interventionally measured PCT.

(6) FIG. 6: Kaplan-Meier plot for patients with high (dashed lined) and low (solid line) levels of pre-interventionally measured hGH.

(7) FIG. 7: Kaplan-Meier plot for patients with high and low levels of pre-interventionally measured MR-proANP and CT-proAVP (pairwise combination of both markers). Solid line: both markers low (below defined threshold); dashed/dotted line: both markers high (above defined threshold); dashed line: first marker low, second marker high; dotted line: first marker high, second marker low.

(8) FIG. 8: Kaplan-Meier plot for patients with high and low levels of pre-interventionally measured MR-proANP and MR-proADM (pairwise combination of both markers). Solid line: both markers low (below defined threshold); dashed/dotted line: both markers high (above defined threshold); dashed line: first marker low, second marker high; dotted line: first marker high, second marker low.

(9) FIG. 9: Kaplan-Meier plot for patients with high and low levels of pre-interventionally measured MR-proANP and CT-proET-1 (pairwise combination of both markers). Solid line: both markers low (below defined threshold); dashed/dotted line: both markers high (above defined threshold); dashed line: first marker low, second marker high; dotted line: first marker high, second marker low.

(10) FIG. 10: Kaplan-Meier plot for patients with high and low levels of pre-interventionally measured MR-proANP and PCT (pairwise combination of both markers). Solid line: both markers low (below defined threshold); dashed/dotted line: both markers high (above defined threshold); dashed line: first marker low, second marker high; dotted line: first marker high, second marker low.

(11) FIG. 11: Kaplan-Meier plot for patients with high and low levels of pre-interventionally measured CT-proAVP and MR-proADM (pairwise combination of both markers). Solid line: both markers low (below defined threshold); dashed/dotted line: both markers high (above defined threshold); dashed line: first marker low, second marker high; dotted line: first marker high, second marker low.

(12) FIG. 12: Kaplan-Meier plot for patients with high and low levels of pre-interventionally measured CT-proAVP and CT-proET-1 (pairwise combination of both markers). Solid line: both markers low (below defined threshold); dashed/dotted line: both markers high (above defined threshold); dashed line: first marker low, second marker high; dotted line: first marker high, second marker low.

(13) FIG. 13: Kaplan-Meier plot for patients with high and low levels of pre-interventionally measured CT-proAVP and PCT (pairwise combination of both markers). Solid line: both markers low (below defined threshold); dashed/dotted line: both markers high (above defined threshold); dashed line: first marker low, second marker high; dotted line: first marker high, second marker low.

(14) FIG. 14: Kaplan-Meier plot for patients with high and low levels of pre-interventionally measured MR-proADM and CT-proET-1 (pairwise combination of both markers). Solid line: both markers low (below defined threshold); dashed/dotted line: both markers high (above defined threshold); dashed line: first marker low, second marker high; dotted line: first marker high, second marker low.

(15) FIG. 15: Kaplan-Meier plot for patients with high and low levels of pre-interventionally measured MR-proADM and PCT (pairwise combination of both markers). Solid line: both markers low (below defined threshold); dashed/dotted line: both markers high (above defined threshold); dashed line: first marker low, second marker high; dotted line: first marker high, second marker low.

(16) FIG. 16: Kaplan-Meier plot for patients with high and low levels of pre-interventionally measured CT-proET-1 and PCT (pairwise combination of both markers). Solid line: both markers low (below defined threshold); dashed/dotted line: both markers high (above defined threshold); dashed line: first marker low, second marker high; dotted line: first marker high, second marker low.

(17) FIG. 17: Kaplan-Meier plot for patients with high and low levels of pre-interventionally measured hGH and MR-proANP (pairwise combination of both markers). Solid line: both markers low (below defined threshold); dashed/dotted line: both markers high (above defined threshold); dashed line: first marker low, second marker high; dotted line: first marker high, second marker low.

(18) FIG. 18: Kaplan-Meier plot for patients with high and low levels of pre-interventionally measured hGH and MR-proADM (pairwise combination of both markers). Solid line: both markers low (below defined threshold); dashed/dotted line: both markers high (above defined threshold); dashed line: first marker low, second marker high; dotted line: first marker high, second marker low.

(19) FIG. 19: Kaplan-Meier plot for patients with high and low levels of pre-interventionally measured hGH and CT-proET-1 (pairwise combination of both markers). Solid line: both markers low (below defined threshold); dashed/dotted line: both markers high (above defined threshold); dashed line: first marker low, second marker high; dotted line: first marker high, second marker low.

(20) FIG. 20: Kaplan-Meier plot for patients with high and low levels of pre-interventionally measured hGH and PCT (pairwise combination of both markers). Solid line: both markers low (below defined threshold); dashed/dotted line: both markers high (above defined threshold); dashed line: first marker low, second marker high; dotted line: first marker high, second marker low.

(21) FIG. 21: Kaplan-Meier plot for patients with high levels of MR-proANP, MR-proADM, CT-proAVP, CT-proET-1 and/or PCT. Upper to lower curve: 0, 1, 2, 3 and 4 or more of the markers above defined threshold.

(22) FIG. 22: Kaplan-Meier plot for patients with high levels of pre-interventionally measured MR-proANP, MR-proADM, CT-proAVP, CT-proET-1 and/or PCT. MR-proANP and CT-proAVP have been counted twice. Upper to lower curve: 0, 1, 2, 3, 4, 5 and 6 or more of the markers above defined threshold.

(23) FIG. 23: Kaplan-Meier plot for patients with high and low levels of pre-interventionally measured MR-proANP and a modified Rankin Scale of 5 on day 5.

(24) FIG. 24: Kaplan-Meier plot for patients with high and low levels of pre-interventionally measured MR-proANP and a Barthel Index <85% on day 5.

(25) FIG. 25: Kaplan-Meier plot for patients with high and low levels of pre-interventionally measured PCT and a NIHSS Index <10 on day 1.

(26) FIG. 26: Kaplan-Meier plot for patients with high and low levels of pre-interventionally measured MR-proANP and a NIHSS Index <10 on day 5.

(27) FIG. 27: Kaplan-Meier plot for patients with high and low levels of pre-interventionally measured PCT and a NIHSS Index <10 on day 1.

(28) FIG. 28: Kaplan-Meier plot for patients with high and low levels of MR-proANP measured pre-interventionally (day 0) and/or day 5. Solid line: level below threshold on day 0 and day 5; dashed line: level below threshold on day 0 and above threshold on day 5; dotted lined: level above threshold on day 0 and below threshold on day 5; dashed/dotted line: level above threshold on day 0 and day 5.

(29) FIG. 29: Kaplan-Meier plot for patients with high and low levels of CT-proAVP measured pre-interventionally (day 0) and/or day 5. Solid line: level below threshold on day 0 and day 5; dashed line: level below threshold on day 0 and above threshold on day 5; dotted lined: level above threshold on day 0 and below threshold on day 5; dashed/dotted line: level above threshold on day 0 and day 5.

(30) FIG. 30: Kaplan-Meier plot for patients with high and low levels of MR-proADM measured pre-interventionally (day 0) and/or day 5. Solid line: level below threshold on day 0 and day 5; dashed line: level below threshold on day 0 and above threshold on day 5; dotted lined: level above threshold on day 0 and below threshold on day 5; dashed/dotted line: level above threshold on day 0 and day 5.

(31) FIG. 31: Kaplan-Meier plot for patients with high and low levels of CT-proET-1 measured pre-interventionally (day 0) and/or day 5. Solid line: level below threshold on day 0 and day 5; dashed line: level below threshold on day 0 and above threshold on day 5; dotted lined: level above threshold on day 0 and below threshold on day 5; dashed/dotted line: level above threshold on day 0 and day 5.

(32) FIG. 32: Kaplan-Meier plot for patients with high and low levels of PCT measured pre-interventionally (day 0) and/or day 5. Solid line: level below threshold on day 0 and day 5; dashed line: level below threshold on day 0 and above threshold on day 5; dotted lined: level above threshold on day 0 and below threshold on day 5; dashed/dotted line: level above threshold on day 0 and day 5.

(33) FIG. 33: Kaplan-Meier plot for patients with high NIHSS (>10) and low NIHSS (<10) on day 5.

(34) FIG. 34: Kaplan-Meier plot for patients with high/low levels of MR-proANP measured on day 5.

(35) FIG. 35: Kaplan-Meier plot for patients with high/low levels of MR-proANP and high/low NIHSS on day 5. Solid line: marker level low, NIHSS low; dashed line: marker level low, NIHSS high; dotted line: marker level high, NIHSS low, dashed/dotted line: marker level high, and NIHSS high.

(36) FIG. 36: Kaplan-Meier plot for patients with high/low levels of CT-proAVP on day 5.

(37) FIG. 37: Kaplan-Meier plot for patients with high/low levels of CT-proAVP and high/low NIHSS on day 5. Solid line: marker level low, NIHSS low; dashed line: marker level low, NIHSS high; dotted line: marker level high, NIHSS low, dashed/dotted line: marker level high, and NIHSS high.

(38) FIG. 38: Kaplan-Meier plot for patients with high/low levels of CT-proET-1 on day 5.

(39) FIG. 39: Kaplan-Meier plot for patients with high/low levels of CT-proET-1 and high/low NIHSS on day 5. Solid line: marker level low, NIHSS low; dashed line: marker level low, NIHSS high; dotted line: marker level high, NIHSS low, dashed/dotted line: marker level high, and NIHSS high.

(40) FIG. 40: Kaplan-Meier plot for patients with Barthel Index <85% (dashed line) and >85% (solid line) on day 5.

(41) FIG. 41: Kaplan-Meier plot for patients with high/low levels of MR-proANP and Barthel Index </>85% on day 5. Solid line: marker level low, Barthel Index >85%; dashed line: marker level low, Barthel Index <85%; dotted line: marker level high, Barthel Index >85%, dashed/dotted line: marker level high, and Barthel Index <85%.

(42) FIG. 42: Kaplan-Meier plot for patients with high/low levels of CT-proAVP and Barthel Index </>85% on day 5. Solid line: marker level low, Barthel Index >85%; dashed line: marker level low, Barthel Index <85%; dotted line: marker level high, Barthel Index >85%, dashed/dotted line: marker level high, and Barthel Index <85%.

(43) FIG. 43: Kaplan-Meier plot for patients with high/low levels of CT-proET-1 and Barthel Index </>85% on day 5. Solid line: marker level low, Barthel Index >85%; dashed line: marker level low, Barthel Index <85%; dotted line: marker level high, Barthel Index >85%, dashed/dotted line: marker level high, and Barthel Index <85%.

(44) FIG. 44: Kaplan-Meier plot for patients with a modified Rankin Scale Score of 0-2 (solid line) and 3-6 (dashed line) on day 5.

(45) FIG. 45: Kaplan-Meier plot for patients with high/low levels of MR-proANP and a modified Rankin Scale Score between 0-2 or 3-6 on day 5. Solid line: marker level low, Rankin 0-2; dashed line: marker level low, Rankin 3-6; dotted line: marker level high, Rankin 0-2, dashed/dotted line: marker level high, and Rankin 3-6.

(46) FIG. 46: Kaplan-Meier plot for patients with high/low levels of MR-proADM and a modified Rankin Scale Score between 0-2 or 3-6 on day 5. Solid line: marker level low, Rankin 0-2; dashed line: marker level low, Rankin 3-6; dotted line: marker level high, Rankin 0-2, dashed/dotted line: marker level high, and Rankin 3-6.

(47) FIG. 47: Kaplan-Meier plot for patients with high/low levels of CT-proAVP and a modified Rankin Scale Score between 0-2 or 3-6 on day 5. Solid line: marker level low, Rankin 0-2; dashed line: marker level low, Rankin 3-6; dotted line: marker level high, Rankin 0-2, dashed/dotted line: marker level high, and Rankin 3-6.

(48) FIG. 48: Kaplan-Meier plot for patients with high/low levels of CT-proET-1 and a modified Rankin Scale Score between 0-2 or 3-6 on day 5. Solid line: marker level low, Rankin 0-2; dashed line: marker level low, Rankin 3-6; dotted line: marker level high, Rankin 0-2, dashed/dotted line: marker level high, and Rankin 3-6.

(49) FIG. 49: Kaplan-Meier plot for patients with high/low levels of PCT and a modified Rankin Scale Score between 0-2 or 3-6 on day 5. Solid line: marker level low, Rankin 0-2; dashed line: marker level low, Rankin 3-6; dotted line: marker level high, Rankin 0-2, dashed/dotted line: marker level high, and Rankin 3-6.

(50) FIG. 50: Box plot of correlation between the modified Rankin Scale determined after 3 months and the level of pre-interventionally measured CT-proAVP (A) and MR-proANP (B), respectively. Rankin 0 to 6 on x-axis from left to right.

(51) FIG. 51: Receiver Operating characteristics (ROC) plot for pre-interventionally measured CT-pro A VP and dichotomized modified Rankin Scale after 3 months (good outcome=mRS 0-2; bad outcome=mRS 3-6). Area under the curve (AUC)=0.723.

(52) FIG. 52: ROC plot for pre-interventionally measured MR-proANP and dichotomized modified Rankin Scale after 3 months (good outcome=mRS 0-2; bad outcome=mRS 3-6). AUC=0.703.

(53) FIG. 53: ROC plot for pre-interventionally measured MR-proADM and dichotomized modified Rankin Scale after 3 months (good outcome=mRS 0-2; bad outcome=mRS 3-6). AUC=0.658.

(54) FIG. 54: ROC plot for pre-interventionally measured CT-proET-1 and dichotomized modified Rankin Scale after 3 months (good outcome=mRS 0-2; bad outcome=mRS 3-6). AUC=0.608.

(55) FIG. 55: ROC plot for pre-interventionally measured PCT and dichotomized modified Rankin Scale after 3 months (good outcome=mRS 0-2; bad outcome=mRS 3-6). AUC=0.580.

(56) FIG. 56: ROC plot for pre-interventionally measured hGH and dichotomized modified Rankin Scale after 3 months (good outcome=mRS 0-2; bad outcome=mRS 3-6). AUC=0.581.

(57) FIG. 57: Distribution of MR-proANP levels in patients with ischemic stroke, hemorrhagic stroke and TIA. Median values are 137.0 pmol/L, 124.0 pmol/L and 90.6 pmol/L, respectively.

(58) FIG. 58: ROC plot for differential diagnosis of ischemic stroke and TIA using MR-proANP as single marker. AUC=0.65 (P<0.0001).

(59) FIG. 59: ROC plot for differential diagnosis of hemorrhagic stroke and TIA using MR-proANP as single marker. AUC=0.65 (P<0.001)

(60) FIG. 60: Distribution of CT-proAVP (copeptin) levels in patients with ischemic stroke, hemorrhagic stroke and TIA. Median values are 11.0 pmol/L, 16.6 pmol/L and 4.8 pmol/L, respectively.

(61) FIG. 61: ROC plot for differential diagnosis of ischemic stroke and TIA using copeptin as single marker. AUC=0.71 (P<0.0001).

(62) FIG. 62: ROC plot for differential diagnosis of hemorrhagic stroke and TIA using copeptin as single marker. AUC=0.74 (P<0.0001)

(63) FIG. 63: Distribution of MR-proADM levels in patients with ischemic stroke, hemorrhagic stroke and TIA. Median values are 0.74 nmol/L, 0.71 nmol/L and 0.61 nmol/L, respectively.

(64) FIG. 64: ROC plot for differential diagnosis of ischemic stroke and TIA using MR-proADM as single marker. AUC=0.61 (P<0.0001).

(65) FIG. 65: ROC plot for differential diagnosis of hemorrhagic stroke and TIA using MR-proADM as single marker. AUC=0.59 (P=0.033)

(66) FIG. 66: Distribution of CT-proET-1 levels in patients with ischemic stroke, hemorrhagic stroke and TIA. Median values are 72.0 pmol/L, 75.7 pmol/L and 63.3 pmol/L, respectively.

(67) FIG. 67: ROC plot for differential diagnosis of ischemic stroke and TIA using CT-proET-1 as single marker. AUC=0.59 (P<0.001).

(68) FIG. 68: ROC plot for differential diagnosis of hemorrhagic stroke and TIA using CT-proET-1 as single marker. AUC=0.68 (P<0.0001)

(69) FIG. 69: Distribution of PCT levels in patients with ischemic stroke, hemorrhagic stroke and TIA. Median values are 0.018 ng/mL, 0.018 ng/mL and 0.016 ng/mL, respectively.

(70) FIG. 70: ROC plot for differential diagnosis of ischemic stroke and TIA using PCT as single marker. AUC=0.58 (P<0.001).

(71) FIG. 71: ROC plot for differential diagnosis of hemorrhagic stroke and TIA using PCT as single marker. AUC=0.56 (P=0.17)

(72) FIG. 72: Distribution of hGH levels in patients with ischemic stroke, hemorrhagic stroke and TIA. Median values are 0.42 ng/mL, 0.41 ng/mL and 0.24 ng/mL, respectively.

(73) FIG. 73: ROC plot for differential diagnosis of ischemic stroke and TIA using hGH as single marker. AUC=0.60 (P<0.001).

(74) FIG. 74: ROC plot for differential diagnosis of hemorrhagic stroke and TIA using hGH as single marker. AUC=0.61 (P=0.013)

(75) FIG. 75: SEQ ID NO:1 (amino acid sequence of pre-pro-ADM)

(76) FIG. 76: SEQ ID NO:2 (amino acid sequence of pro-ADM)

(77) FIG. 77: SEQ ID NO:3 (amino acid sequence of pro-ADM N20)

(78) FIG. 78: SEQ ID NO:4 (amino acid sequence of MR-pro-ADM)

(79) FIG. 79: SEQ ID NO:5 (amino acid sequence of ADM)

(80) FIG. 80: SEQ ID NO:6 (amino acid sequence of pre-pro-ANP)

(81) FIG. 81: SEQ ID NO:7 (amino acid sequence of pro-ANP)

(82) FIG. 82: SEQ ID NO:8 (amino acid sequence of ANP)

(83) FIG. 83: SEQ ID NO:9 (amino acid sequence of NT-proANP)

(84) FIG. 84: SEQ ID NO:10 (amino acid sequence of amino acids 53-90 of proANP)

(85) FIG. 85: SEQ ID NO:11 (amino acid sequence of pre-pro-AVP)

(86) FIG. 86: SEQ ID NO:12 (amino acid sequence of pro-AVP)

(87) FIG. 87: SEQ ID NO:13 (amino acid sequence of AVP)

(88) FIG. 88: SEQ ID NO:14 (amino acid sequence of CT-pre-proAVP or Copeptin)

(89) FIG. 89: SEQ ID NO:15 (amino acid sequence of Neurophysin II)

(90) FIG. 90: SEQ ID NO:16 (amino acid sequence of pre-pro-ET-1)

(91) FIG. 91: SEQ ID NO:17 (amino acid sequence of pro-ET-1)

(92) FIG. 92: SEQ ID NO:18 (amino acid sequence of ET-1)

(93) FIG. 93: SEQ ID NO:19 (amino acid sequence of CT-pro-ET-1)

(94) FIG. 94: SEQ ID NO:20 (amino acid sequence of Big-ET-1)

(95) FIG. 95: SEQ ID NO:21 (amino acid sequence of pre-pro-Calcitonin)

(96) FIG. 96: SEQ ID NO:22 (amino acid sequence of PCT)

(97) FIG. 97: SEQ ID NO:23 (amino acid sequence of N-terminal PCT)

(98) FIG. 98: SEQ ID NO:24 (amino acid sequence of Calcitonin)

(99) FIG. 99: SEQ ID NO:25 (amino acid sequence of Katacalcin)

(100) FIG. 100: SEQ ID NO:26 (amino acid sequence of pre-pro-BNP)

(101) FIG. 101: SEQ ID NO:27 (amino acid sequence of pro-BNP)

(102) FIG. 102: SEQ ID NO:28 (amino acid sequence of NT-pro-BNP)

(103) FIG. 103: SEQ ID NO:29 (amino acid sequence of BNP

EXAMPLES

Example 1: Clinical Study

(104) Study Setting, Inclusion/Exclusion Criteria

(105) The study was set at the emergency and neurological and neurosurgical clinic of the University Hospital of Basel. All consecutive patients who are admitted to the emergency department with an ischemic or hemorrhagic stroke or transient ischemic attack (TIA) according to the World Health Organization criteria with symptom onset within the last 3 days were included into the study. Patients without an informed consent were excluded.

(106) Baseline Data Collection

(107) Access to data of all eligible patients that are not included into this study is important to avoid a selection bias. Thus, baseline data and information on inclusion and exclusion criteria in all eligible patients were collected irrespective whether they are or are not included into the study. This allows the comparison of baseline data of eligible patients who consented to participate with those who did not. Baseline data collection in patients were collected by the investigators and comprised:

(108) a) age

(109) b) gender

(110) c) BMI

(111) d) Medical history items: actual history that preceded the hospitalization; ABCD score (Rothwell et al., 2005. A simple score (ABCD) to identify individuals at high early risk of stroke after transient ischaemic attack. Lancet 366: 29-36) in patients with transient ischemic attack; family history; relevant co-morbidities also assessed by the charlson index (Goldstein et al., 2004. Charlson Index comorbidity adjustment for ischemic stroke outcome studies. Stroke 35: 1941-5) (i.e. hypertension, previous stroke, previous TIA, ischemic heart disease, atrial fibrillation, diabetes mellitus, renal and liver dysfunction, congestive heart failure, dyslipidemia; comorbidities with the risk of hyponatremia (severe hypothyroidism, glucocorticoid insufficiency, neoplasm, HIV infection); smoking history (pack-years) and status (pack per day); current medication; alcohol consumption (glass and grams per day); time from onset of symptoms to admission.

(112) e) Place of residence: i.e. independent living, defined as living at home or in an old people's home with or without support of the family circle and/or professional care (the family circle consists of the spouse and/or other important persons who live together with the patient; dependent living, defined as nursing home long-stay department, other hospital.

(113) f) Clinical items: physical examination including neurological status, NIHSS (to assess the severity of stroke) and Glasgow Coma scale (GCS; Adams et al., 1999. Baseline NIH Stroke Scale score strongly predicts outcome after stroke: A report of the Trial of Org 10172 in Acute Stroke Treatment (TOAST). Neurology 53: 126-31), blood pressure, pulse rate, weight, volume status (including skin turgor, jugular venous distension, auscultation, if available flow sheet of fluid intake and loss), body temperature; in neurosurgical patients intracerebral pressure if performed within the routine clinical management.

(114) g) Clinical symptoms of hyponatraemia were evaluated on admission and in case of sodium imbalance in neurological patients. In patients undergoing intracranial surgery evaluate clinical symptoms were evaluated daily. Specifically the presence of headache, anorexia, nausea, vomiting, muscle cramps and aches, seizures, confusion, apathetic or lethargic development was monitored.

(115) h) Routine/Standard laboratory tests: routine blood sampling including: hematocrit, blood urea nitrogen, bicarbonate, total protein, albumin, uric acid serum and urine electrolytes, urine and serum osmolality, creatinine, lipids, TSH, fT4, T3, and basal cortisol. All blood sampling was done before any food intake, or smoking, if feasible. Alternatively, influencing factors were monitored.

(116) i) Imaging: Computer tomography or MRI of the neurocranium (T1, T2, diffusion-weighted image sequence, with or without contrast), if indicated magnetic resonance angiography or conventional cerebral angiography. The time-points of contrast agent application were recorded.

(117) Stroke patients were also classified on the basis of the vascular territory of the ischemic lesion as follows: total anterior circulation syndrome (TACS), partial anterior circulation syndrome (PACS), lacunar circulation syndrome (LACS), posterior circulation syndrome (POCS).

(118) j) Further investigations: Stroke patients had neurosonography, echocardiography, standard 12-leaf electrocardiography and 24-hour electrocardiography and then were classified by etiology of strokes according to Trial of Org 10172 in acute Treatment (TOAST) stroke subtype classification, which differs between large artery atherosclerosis, cardioembolism, small-artery occlusion, other etiology, and undetermined etiology.

(119) Informed Consent Statement

(120) The study was approved by the ethics committee of Basel (Ethikkommission beider Basel). It is important to note that this is an exploratory and observational study; the only study related intervention will be the asseveration of 7.5 ml of plasma obtained during the routinely performed blood sampling. Therefore, patients were required to provide written informed consent that they agree for the use of their data for scientific purposes. In patients, in which informed consent was not feasible due to sequela of the acute CNS lesion (the latter a prerequisite for inclusion), patients' next to kin could sign an assent form to state the presumptive will of the patient. In case, next of kin were not readily available, a treating physicianwho must not be involved in the studyhad to certify that there are no objections for inclusion in the study from his point of view. Only after these informed consent procedures the patient could be included in the study.

(121) Management of Participants Throughout the Trial

(122) Step 1.

(123) All eligible patients in the emergency department or the neurological ward were included into the study.

(124) Step 2.

(125) All baseline data were collected.

(126) Step 3.

(127) During hospitalization clinical items including weight, blood pressure, pulse rate, volume status and body temperature were assessed by chart review until discharge. Fluid treatment and drugs Potential symptoms of hyponatremia, i.e. headache, nausea, vomiting, muscle cramps and aches, anorexia, impaired consciousness, seizure. Routinely performed laboratory tests (chemogram, plasma glucose, serum osmolality, urine osmolality, sodium in urine, hematocrit) were sampled at the time-points upon routine blood sampling.

(128) Step 4.

(129) In all patients, on day 5 of the hospitalization, a clinical examination with NIHSS, Barthel Index and Rankin Scale were performed (Collin et al., 1988. The Barthel ADL Index: a reliability study, International Disability Study 10: 61-3; Bonita and Beaglehole, 1988. Modification of Rankin Scale: Recovery of motor function after stroke. Stroke 19:1497-1500).

(130) The future place of residence (i.e. dependent vs. independent living) was assessed.

(131) Step 6.

(132) In patients with ischemic stroke a telephone follow-up regarding morbidity and mortality (as assessed by the Barthel Index and Rankin Scale) was obtained after 3 months. An unfavorable outcome was defined as a Barthel index <85 or modified Rankin scale of 3 to 6.

(133) Measurement of Marker Peptide Concentrations

(134) MR-proANP levels were measured with a chemiluminescence sandwich immunoassay with a lower detection limit of 6 pmol/L. In 325 healthy individuals the range of MR-proANP concentrations was 9.6-313 pmol/L with a median of 45 pmol/L. Concentrations differed not significantly between males and females, but significantly correlated with the age of the subjects.

(135) MR-proADM levels were measured with a chemiluminescence sandwich immunoassay with a lower detection limit of 0.08 nmol/L. In 264 healthy individuals (117 male and 147 female) MR-proADM values followed a Gaussian distribution with mean (SD) values of 0.33 (0.07) nmol/L and a range of 0.10-0.64 nmol/L. There was no significant difference between gender, but MR-proADM concentrations significantly correlated with age.

(136) CT-proET-1 levels were measured with a chemiluminescence sandwich immunoassay with a lower detection limit of 0.4 pmol/L. In 326 healthy individuals (150 male and 176 female) CT-proET-1 values followed a Gaussian distribution with a mean (SD) of 44.3 (10.6) pmol/L and a range of 10.5-77.4 pmol/L. Mean CT-proET-1 concentrations in males and females were not significantly different but significantly correlated with age.

(137) CT-proAVP (Copeptin) levels were measured with a chemiluminescence sandwich immunoassay with a lower detection limit of 1.7 pmol/L. In 359 healthy individuals (153 men and 206 women), median CT-proAVP levels were 4.2 pmol/L ranging from 1.0-13.8 pmol/L. Median concentrations of CT-proAVP differed significantly between male and female. There was no correlation between CT-proAVP levels and age.

(138) PCT levels were measured with the chemiluminescence sandwich immunoassay PCT sensitive (B.R.A.H.M.S, Hennigsdorf, Germany) with a lower detection limit of 7 pg/ml. In 500 healthy individuals, the median was 13.5 pg/ml (range <7 to 63 pg/ml). There were no significant differences in the range and median PCT values between males and females or among age groups.

(139) Human growth hormone was measured with a newly developed chemiluminescence sandwich immunoassay for the specific detection of the most abundant 22 kDa hGH isoform. The median concentration of the 22 kDa hGH isoform in a cohort of 50 blood donors was 0.19 ng/ml (range 0.05-8.82 ng/ml).

(140) The plasma levels of the marker peptides MR-proANP, MR-proADM, CT-proET-1, CT-proAVP (Copeptin), PCT and hGH have been determined in 352 patients with ischemic stroke, in 32 patients with cerebral hemorrhage and in 102 patients with TIA. Blood samples were taken at the day of hospitalization (day 0) and 1, 3 and 5 days after hospitalization.

Examples 2 to 11

(141) Examples 2 to 11 concern the same group of patients and are based on the study of example 1. Tables 1 and 2 summarize the patients of the study and their outcome. Blood samples (EDTA treated plasma samples) were taken from a group of 501 stroke or TIA patients. The group of patients is the same as in example 1, however, the number may slightly differ, since not for all days all patients were available and not on all days enough sample volume was available for every patient to determine all markers.

Example 2: Prognosis of Survival (Death within 3 Months) with Markers Measured on Day 0

(142) The outcome for patients with stroke or TIA within 3 (4) months after the stroke or TIA is investigated and correlated to the level of the markers MR-proANP, MR-proADM, CT-proAVP, CT-proET1 and PCT in samples of said patients. Samples have been taken on day 0 (day of hospitalization). From this correlation hazard ratios (HR) have been calculated for the given cut-off values (threshold values). The cut-off values have been determined by maximizing the sum of sensitivity and specificity as determined from time-dependent ROC analysis.

(143) Tables 3a to 4 summarize the HR values for the different markers and their combinations. FIGS. 1 to 27 show the Kaplan-Meier plots (proportion of patients surviving within four months) for the different markers/marker combinations and in combination with Rankin/Barthels Index/NIHSS.

(144) TABLE-US-00002 TABLE 1 Summary of patients definite diagnosis frequency percentage hemorrhagic stroke 32 6.4 ischemic stroke 362 72.3 TIA 107 21.4 sum 501 100.0

(145) TABLE-US-00003 TABLE 2 Survival of patients dead within 3 months frequency percentage yes 53 10.6 no 447 89.2 unknown 1 0.2 sum 501 100.0

(146) TABLE-US-00004 TABLE 3a Hazard ratios (HR) for different markers Marker HR cut off MR-proANP 10.8 188 CT-proAVP 9.4 20.257 MR-proADM 4.7 0.666 CT-proET-1 4.1 98.3 PCT sens 3.4 0.026 hGH 1.4 0.34

(147) TABLE-US-00005 TABLE 3b Hazard ratios (HR) for different marker combinations HR (both high Marker combination vs both low) MR-proANP + CT-proAVP 61.4 MR-proANP + MR-proADM 62.7 MR-proANP + CT-proET-1 17.6 MR-proANP + PCT sens 25.2 CT-proAVP + MR-proADM 37.9 CT-proAVP + CT-proET-1 13.2 CT-proAVP + PCT sens 25.3 MR-proADM + CT-proET-1 8.7 MR-proADM + PCT sens 11.4 CT-proET-1 + PCT sens 6.8 HGH + MR-proANP 12.3 HGH + MR-proADM 5.8 HGH + CT-proET-1 5.0 HGH + PCT sens 4.4

(148) TABLE-US-00006 TABLE 3c Hazard ratios (HR) for combination of all six markers HR (>3 high vs Marker combinations all low) Combination of 6, model 1 >100 Combination of 6, model 2 >100

(149) TABLE-US-00007 TABLE 4 Hazard ratios (HR) of different markers in combination with different scores HR HR HR HR HR com- mRS, Barthel, NIHSS, NIHSS, Marker plete day 5 = 5 day 5 <85% day 1 >10 day 5 >10 MR-proANP 10.8 22.4 31.1 7.8 >100 CT-proAVP 9.4 1.8 3.5 2.9 3.0 MR-proADM 4.7 2.3 1.4 3.0 3.2 CT-proET-1 4.1 2.4 2.1 3.2 3.2 PCT sens 3.4 2.5 2.5 4.3 6.8

Example 3: Prognosis of Survival (Death within 3 Months) with Markers Measured on Day 0 and Day 5 (Days 0 and 5 in Combination)

(150) TABLE-US-00008 TABLE 5 Hazard ratios (HR) for different markers HR HR (both high Marker, day 0 and day 5 (day 0 only) vs both low) cut off MR-proANP 10.8 26.5 188 CT-proAVP 9.4 24.2 20.257 MR-proADM 4.7 6.6 0.666 CT-proET-1 4.1 8.7 98.3 PCT sens 3.4 7.8 0.026 hGH 1.4 1.4 0.34

(151) Table 5 summarizes the HR values for the different markers in samples taken on day 0 and 5. FIGS. 28 to 32 show the Kaplan-Meier plots (proportion of patients surviving within four months) for the different markers.

Example 4: Prognosis of Survival (Death within 3 Months) with Markers Measured on Day 5 and in Combination with the NIHSS

(152) TABLE-US-00009 TABLE 6 Hazard ratios (HR) for different markers Marker, day 5 and HR HR (both high NIHSS (>10) (day 5 only) vs both low) cut off NIHSS 7.3 10 MR-proANP 8.0 44.6 188 CT-proAVP 18.3 43.4 20.257 MR-proADM 2.8 14.6 0.666 CT-proET-1 7.9 48.4 98.3 PCT sens 4.1 17.7 0.026 hGH 1.4 6.6 0.34

(153) Table 6 summarizes the HR values for the different markers in samples taken on day 5 and in combination with the NIHSS values for said patients. FIGS. 33 to 39 show the Kaplan-Meier plots (proportion of patients surviving within four months) for NIHSS alone and for the different markers in combination with NIHSS based on samples taken on day 5.

Example 5: Prognosis of Survival (Death within 3 Months) with Markers Measured on Day 5 and in Combination with the Barthel Index

(154) TABLE-US-00010 TABLE 7 Hazard ratios (HR) for different markers Marker, day 5 and HR HR (both high Barthel Index (<85%) (day 5 only) vs both low) cut off Barthel Index 12.3 85% MR-proANP 8.0 44.0 188 CT-proAVP 18.3 >100 20.257 MR-proADM 2.8 14.6 0.666 CT-proET-1 7.9 68.5 98.3 PCT sens 4.1 24.4 0.026 hGH 1.4 7.2 0.34

(155) Table 7 summarizes the HR values for the different markers in samples taken on day 5 and in combination with Barthel Index values for said patients. FIGS. 40 to 43 show the Kaplan-Meier plots (proportion of patients surviving within four months) for Barthel Index for the different markers in combination with Barthel Index based on samples taken on day 5.

Example 6: Prognosis of Survival with Markers Measured on Day 5 and in Combination with the Modified Rankin Scale, Outcome Dead within 3 Months

(156) TABLE-US-00011 TABLE 8 Hazard ratios (HR) for different markers Marker, day 5 and HR HR (both high Rankin (>2) (day 5 only) vs both low) cut off Rankin 10.7 0-2 vs. 3-6 MR-proANP 8.0 37.1 188 CT-proAVP 18.3 >100 20.257 MR-proADM 2.8 12.9 0.666 CT-proET-1 7.9 49.6 98.3 PCT sens 4.1 20.4 0.026 hGH 1.4 5.7 0.34

(157) Table 8 summarizes the HR values for the different markers in samples taken on day 5 and in combination with the modified Rankin Scale values for said patients. FIGS. 44 to 49 show the Kaplan-Meier plots (proportion of patients surviving within four months) for the modified Rankin Scale alone and for the different markers in combination with the modified Rankin Scale based on samples taken on day 5.

Example 7: Prognosis of Re-Stroke/Re-TIA (within 3 Months) with Markers Measured on Day 0

(158) Table 9 summarizes the re-occurrence of strokes (re-stroke) and TIAs (re-TIA) in the group of patients. Tables 10 to 14 summarize the calculated HR values for the different markers and marker combinations.

(159) TABLE-US-00012 TABLE 9 re-stroke or re-TIA within 3 months re-event within 3 months frequency percentage no 419 83.6 yes 29 5.8 unknown 53 10.6 sum 501 100.0

(160) TABLE-US-00013 TABLE 10 Odd ratios (OR) for different markers OR Marker OR (TIA only) cut off MR-proANP 3.6 21.7 323.2 CT-proAVP 3.0 >100 38.6 MR-proADM 3.7 4.2 1.18 CT-proET-1 7.0 59.2 PCT sens 2.9 3.2 0.057 hGH 1.9 0.34

(161) TABLE-US-00014 TABLE 11 Odd ratios for different marker combinations OR (both high OR (both high vs both low) Marker (combination) vs both low) TIA only MR-proANP + CT-proAVP 7.4 >100 MR-proANP + MR-proADM 8.1 >100 MR-proANP + CT-proET-1 76.0 MR-proANP + PCT sens 4.5* 24.0* CT-proAVP + MR-proADM 10.5 >100 CT-proAVP + CT-proET-1 >100 CT-proAVP + PCT sens 3.5* >100* MR-proADM + CT-proET-1 15.2 MR-proADM + PCT sens 5.1* CT-proET-1 + PCT sens 18.5 hGH + MR-proADM 4.9 *not enough patients in the group both high to allow for calculation of OR; however, an OR is given for both low vs. high in first marker.

(162) Table 13a to 13e: Calculation of Odd Ratios for all Patients (Re-Stroke and Re-TIA)

(163) TABLE-US-00015 TABLE 13a Calculation of OR for MR-proANP Re-Stroke or Re-TIA Marker no yes sum MR-proANP <cut off 360 20 380 >cut off 35 7 42 sum 395 27 422 Odds Ratio 3.6

(164) TABLE-US-00016 TABLE 13b Calculation of OR for CT-proAVP Re-Stroke or Re-TIA Marker no yes sum CT-proAVP <cut off 346 19 365 >cut off 48 8 56 sum 394 27 421 Odds Ratio 3.0

(165) TABLE-US-00017 TABLE 13c Calculation of OR for MR-proADM Re-Stroke or Re-TIA Marker no yes sum MR-proADM <cut off 372 22 394 >cut off 23 5 28 sum 395 27 422 Odds Ratio 3.7

(166) TABLE-US-00018 TABLE 13d Calculation of OR for combination of MR-proANP and CT-proAVP Re-Stroke or Re-TIA Marker model no yes sum MR-proANP + both low 325 17 342 CT-proAVP both high 13 5 18 sum 338 22 360 Odds Ratio 7.4 MR-proANP + both low 346 19 365 MR-proADM both high 9 4 13 sum 355 23 378 Odds Ratio 8.1

(167) TABLE-US-00019 TABLE 13e Calculation of OR for combination of CT-proAVP and MR-proADM Re-Stroke or Re-TIA Marker model no yes sum CT-proAVP + both low 331 18 349 MR-proADM both high 7 4 11 sum 338 22 360 Odds Ratio 10.5

(168) Table 14a to 14d: Calculation of Odd Ratios for TIA Patients (Re-Stroke and Re-TIA)

(169) TABLE-US-00020 TABLE 14a Calculation of OR for MR-proANP Re-Stroke or Re-TIA Marker no yes sum MR-proANP <cut off 87 8 95 >cut off 1 2 3 sum 88 10 98 Odds Ratio 21.8

(170) TABLE-US-00021 TABLE 14b Calculation of OR for CT-proAVP Re-Stroke or Re-TIA Marker no yes sum CT-proAVP <cut off 88 8 96 >cut off 0 2 2 sum 88 10 98 Odds Ratio >100

(171) TABLE-US-00022 TABLE 14c Calculation of OR for MR-proADM Re-Stroke or Re-TIA Marker no yes sum MR-proADM <cut off 83 8 91 >cut off 5 2 7 sum 88 10 98 Odds Ratio 4.2

(172) TABLE-US-00023 TABLE 14d Calculation of OR for CT-proET1 Re-Stroke or Re-TIA Marker no yes sum CT-proET1 <cut off 38 1 39 >cut off 49 9 58 sum 87 10 97 Odds Ratio 7.0

(173) TABLE-US-00024 TABLE 14e Calculation of OR for combination of MR-proANP and CT-proAVP Re-Stroke or Re-TIA Marker model no yes sum MR-proANP + both low 87 7 94 CT-proAVP both high 0 1 1 sum 87 8 95 Odds Ratio >100

(174) TABLE-US-00025 TABLE 14f Calculation of OR for combination of CT-proET1 and CT-proAVP Re-Stroke or Re-TIA Marker model no yes sum CT-proET1 both low 38 1 39 CT-proAVP both high 0 2 2 sum 38 3 41

(175) TABLE-US-00026 TABLE 14g Calculation of OR for combination of MR-proANP and MR-proADM Re-Stroke or Re-TIA Marker model no yes sum MR-proANP both low 82 8 90 MR-proADM both high 0 2 2 sum 82 10 92 Odds Ratio >100

Example 8: Prognosis of Re-Stroke/Re-TIA (within 3 Months) with Markers Measured on Day 0 and in Combination with Day 5

(176) Tables 15 to 16d summarize the results of a ROC analysis (monitoring), particularly the odds ratios, for different markers/Marker combinations.

(177) TABLE-US-00027 TABLE 15 Calculation of OR for different markers OR OR (both high Marker, day 0 and day 5 (only day 0) vs both low) cut off MR-proANP 3.6 5.1 323.2 CT-proAVP 3.0 8.5 38.6 MR-proADM 3.7 4.8 1.18 CT-proET-1 59.2 PCT sens 2.9 4.2 0.057 hGH 4.2 0.34

(178) Table 16a to d: Calculation of OR for single markers, day 0 and day 5:

(179) TABLE-US-00028 TABLE 16a Calculation of OR for MR-proANP Re-Stroke or Re-TIA MR-proANP no yes sum day 0, day 5 both low 241 14 255 both high 17 5 22 sum 258 19 277 Odds Ratio 5.1

(180) TABLE-US-00029 TABLE 16b Calculation of OR for CT-proAVP Re-Stroke or Re-TIA CT-proAVP no yes sum day 0, day 5 both low 220 13 233 both high 4 2 6 sum 224 15 239 Odds Ratio 8.5

(181) TABLE-US-00030 TABLE 16c Calculation of OR for MR-proADM Re-Stroke or Re-TIA MR-proADM no yes sum day 0, day 5 both low 241 15 256 both high 10 3 13 sum 251 18 269 Odds Ratio 4.8

(182) TABLE-US-00031 TABLE 16d Calculation of OR for PCT Re-Stroke or Re-TIA PCT no yes sum day 0, day 5 both low 235 16 251 both high 7 2 9 sum 242 18 260 Odds Ratio 4.2

Example 9: Prognosis of Functional Outcome (mRS, after 3 Months) with Markers Measured on Day 0

(183) Table 17 summarizes the outcome for the group of patients in terms of the modified Rankin Scale (0 to 6). Table 18 lists the Kruskal-Wallis Chi.sup.2 statistic and p value for the correlation of the different markers with the modified Rankin Scale. Table 19 summarizes the outcome in two classes: Patients with a good outcome (Rankin 0-2) and patients with a bad outcome (Rankin 3-6). Tables 20a to 20dc summarize the OR on day 0 of the markers/marker combinations for patients with good or bad outcome after 3 months]. Tables 21a to 21h summarize the calculated odds ratios (OR) for the different markers/marker combinations. FIG. 50 illustrates the correlation between Rankin and level of CT-proA VP (A) and MRproANP (B), respectively, in a box plot. FIGS. 51 to 56 illustrates the ROC plots for the different markers.

(184) TABLE-US-00032 TABLE 17 Frequency distribution for the outcome (measured with the modified Rankin Scale) of Stroke/TIA patients after 3 months Rankin after 3 months frequency percentage 0 155 30.9 1 106 21.2 2 53 10.6 3 55 11.0 4 56 11.2 5 20 4.0 6 53 10.6 unknown 3 0.6 sum 501 100.0

(185) TABLE-US-00033 TABLE 18 Results for different markers Ranking 3 months (0-6) Chi.sup.2 p-value MR-proANP 80.4 <0.0001 CT-proAVP 87.2 <0.0001 MR-proADM 40.5 <0.0001 CT-proET-1 26.0 0.0005 PCT sens 22.2 0.0024

(186) TABLE-US-00034 TABLE 19 Rankin after 3 months Rankin after 3 months frequency percentage Rankin 0-2 314 62.7 (good) Rankin 3-6 184 36.7 (bad) unknown 3 0.6 sum 501 100.0

(187) TABLE-US-00035 TABLE 20a OR for different markers Marker OR cut off MR-proANP 4.5 188 CT-proAVP 5.1 20.257 MR-proADM 2.8 0.666 CT-proET-1 3.6 98.3 PCT sens 2.1 0.026 hGH 1.5 0.34

(188) TABLE-US-00036 TABLE 20b OR for different marker combinations OR (both high vs Marker (combinations) both low) MR-proANP + CT-proAVP 15.6 MR-proANP + MR-proADM 7.3 MR-proANP + CT-proET-1 7.6 MR-proANP + PCT sens 8.2 CT-proAVP + MR-proADM 9.8 CT-proAVP + CT-proET-1 8.5 CT-proAVP + PCT sens 7.1 MR-proADM + CT-proET-1 5.4 MR-proADM + PCT sens 4.4 CT-proET-1 + PCT sens 4.6 HGH + CT-proAVP 5.6 HGH + MR-proADM 4.3 hGH + CT-proET-1 4.4 hGH + PCT sens 2.7 hGH + MR-proANP 6.2

(189) TABLE-US-00037 TABLE 20c OR for combination of all 6 markers OR (all high vs Marker (combinations) all low combination of 6 22.0

(190) TABLE-US-00038 TABLE 21a Calculation of OR for CT-proAVP Rankin Marker 0-2 3-6 sum CT-proAVP <cut off 246 85 331 >cut off 49 87 136 sum 295 172 467 Odds Ratio 5.1

(191) TABLE-US-00039 TABLE 21b Calculation of OR for MR-proANP Rankin Marker 0-2 3-6 sum MR-proANP <cut off 232 77 309 >cut off 64 95 159 sum 296 172 468 Odds Ratio 4.5

(192) TABLE-US-00040 TABLE 21c Calculation of OR for MR-proADM Rankin Marker 0-2 3-6 sum MR-proADM <cut off 175 59 234 >cut off 121 113 234 sum 296 172 468 Odds Ratio 2.8

(193) TABLE-US-00041 TABLE 21d Calculation of OR for CT-proET-1 Rankin Marker 0-2 3-6 sum CT-proET-1 <cut off 263 121 384 >cut off 30 49 79 sum 293 170 463 Odds Ratio 3.6

(194) TABLE-US-00042 TABLE 21e Calculation of OR for PCT Rankin Marker 0-2 3-6 sum PCT <cut off 238 114 352 >cut off 57 57 114 sum 295 171 466 Odds Ratio 2.1

(195) TABLE-US-00043 TABLE 21f Calculation of OR for combination of MR-proANP and CT-proAVP Rankin Marker Model no yes sum MR-proANP + both low 198 54 252 CT-proAVP both high 15 64 79 sum 213 118 331 Odds Ratio 15.6

(196) TABLE-US-00044 TABLE 21g Calculation of OR for combination of MR-proANP and MR-proADM Rankin Marker Model no yes sum MR-proANP + both low 151 39 190 MR-proADM both high 40 75 115 sum 191 114 305 Odds Ratio 7.3

(197) TABLE-US-00045 TABLE 21h Calculation of OR for combination of MR-proADM and CT-proAVP Rankin Marker Model no yes sum MR-proADM + both low 154 36 190 CT-proAVP both high 28 64 92 sum 182 100 282 Odds Ratio 9.8

Example 10: Prognosis of Functional Outcome (mRS, after 3 Months) with Markers Measured on Day 0 and in Combination with Day 5

(198) Table 22 summarizes the OR for the single markers on day 0 and in combination with day 5. Tables 23 and 24 summarize the calculated odds ratios (OR) for two different markers in correlation with Rankin values.

(199) TABLE-US-00046 TABLE 22 Odd ratios for different markers OR OR (both high Marker, day 0 and day 5 (only day 0) vs both low) cut off MR-proANP 4.5 188 CT-proAVP 5.1 6.5 20.257 MR-proADM 2.8 0.666 CT-proET-1 3.6 98.3 PCT sens 2.1 3.3 0.026

(200) TABLE-US-00047 TABLE 23 Calculation of OR for CT-proAVP, day 0 and day 5 Ranking CT-proAVP 0-2 3-6 sum day 0, day 5 both low 151 54 205 both high 12 28 40 sum 163 82 245 Odds Ratio 6.5

(201) TABLE-US-00048 TABLE 24 Calculation of OR for PCT, day 0 and day 5 Ranking PCT 0-2 3-6 sum day 0, day 5 both low 129 42 171 both high 29 31 60 sum 158 73 231 Odds Ratio 3.3

Example 11: Differential Diagnosis of TIA, Ischemic Stroke and Hemorrhagic Stroke

(202) FIGS. 57 to 74 summarize the results for the use of MR-proANP, copeptin, MR-proADM, CT-proET-1, PCT and hGH as single markers for the differential diagnosis of TIA, ischemic stroke and hemorrhagic stroke. Marker levels have been determined as described above for day 0 and day 1. Median values have been determined for each marker and patient group (hemorrhagic stroke, ischemic stroke, TIA) and ROC curves for the differential diagnosis of hemorrhagic stroke from TIA and ischemic stroke from TIA have been created.

(203) TABLE-US-00049 TABLE 25 Optimal cut-off values for the differential diagnosis of TIA from stroke (ischemic and hemorrhagic stroke, respectively) using single markers. Cut off TIA vs. Cut off TIA vs. Marker Ischemic stroke hemorrhagic stroke CT-proAVP 9.65 pmol/L 11.35 pmol/L CT-proET 67.15 pmol/L 66.35 pmol/L hGH 0.13 ng/ml 0.13 ng/ml MR-proADM 0.67 nmol/L 0.36 nmol/L MR-proANP 130.5 pmol/L 96.1 pmol/L PCT 0.0235 ng/mL 0.0235 ng/mL