SWATH-MS for prospective identification of protein blood biomarkers of rtPA-associated intracranial hemorrhage in acute ischemic stroke: a pilot study.

Intravenous recombinant tissue plasminogen activator (rtPA) is, besides mechanical thrombectomy, the highest class evidence based reperfusion treatment of acute ischemic stroke (AIS). The biggest concern of the therapy is symptomatic intracranial hemorrhage (sICH), which occurs in 3-7% of all treated patients, and is associated with worse functional outcome. Finding a method of the powerful identification of patients at highest risk of sICH, in order to increase the percentage of stroke patients safely treated with rtPA, is one of the most important challenges in stroke research. To address this problem, we designed a complex project to identify blood, neuroimaging, and clinical biomarkers combined for prospective assessment of the risk of rtPA-associated ICH. In this paper we present results of blood proteomic and peptide analysis of pilot 41 AIS patients before rtPA administration (the test ICH group, n = 9 or the controls, without ICH, n = 32). We demonstrated that pre-treatment blood profiles of 15 proteins differ depending on whether the patients develop rtPA-associated ICH or not. SWATH-MS quantification of serum or plasma proteins might allow for robust selection of blood biomarkers to increase the prospective assessment of rtPA-associated ICH over that based solely on clinical and neuroimaging characteristics.

New Remote Cerebral Microbleeds on T2*-Weighted Echo Planar MRI After Intravenous Thrombolysis for Acute Ischemic Stroke.

BACKGROUND: The main and well-defined complication of intravenous administration of recombinant tissue plasminogen activator (tPA) in patients with acute ischemic stroke (AIS) is symptomatic intracranial hemorrhage (sICH). However, rtPA might also be connected with the formation of cerebral microbleeds (CMBs), located remotely from the ischemic lesions, that may remain clinically silent. This association might be important because the load of CMBs has been associated with cognitive impairment. We investigated whether administration of rtPA in AIS results in the appearance of new CMBs and if the initial load of CMBs is associated with hemorrhagic transformation. METHODS: A total of fifty-nine consecutive patients with AIS treated with rtPA underwent MRI including T2*-weighted Echo Planar Imaging (T2*-EPI) shortly before and 7-9 days after rtPA administration. We calculated the load of new CMBs located outside the MR diffusion restriction area in the follow-up imaging and assessed hemorrhagic transformation with ECASS-II scoring. RESULTS: A total of forty-nine patients were included for the final analysis. On initial T2*-EPI-GRE, 37 baseline microbleeds (CMBs) were observed in 14 patients (28.6%). On follow-up T2*-EPI-GRE amount of CMBs increased to a total number of 103. New CMBs were found in 5 (14.3%) of 35 patients without and in 9 (64.3%) of 14 with any baseline CMBs. Multiple logistic regression analysis indicated that presence of baseline CMBs (risk ratio [RR] 5.95, 95% CI 2.69-13.20, p < 0.001) and lower platelets level (risk ratio [RR] 0.992, 95% CI 0.986-0.998, p = 0.007) were independently associated with new CMBs. The baseline load of CMBs was not associated with the risk of hemorrhagic transformation. CONCLUSION: In this study, new CMBs were found in nearly 30% of patients with AIS on the 7-9 days after rtPA treatment. Baseline CMBs correlated with a higher risk of new CMBs appearing after the rtPA treatment, independently of other factors. At the same time, in our sample, baseline CMBs did not correlate with an increased risk of hemorrhagic transformation. Since the associations between the CMBs load and cognitive impairment have already been proved, further studies are warranted to investigate possible associations between the thrombolytic treatment of patients with AIS, mainly those with baseline CMBs, and the risk of earlier cognitive decline.

CXCR7+ and CXCR4+ stem cells and neuron specific enolase in acute ischemic stroke patients.

Stroke causes an efflux of various groups of progenitor/stem cells from bone marrow to bloodstream and a rise in neuron specific enolase (NSE) serum concentrations. The aim of this study was to identify activity of chosen stem/progenitor cells during first 7 days after stroke through correlations between these cells levels and NSE values. Additional goal was to confirm the role of NSE as a prognostic marker of ischemic stroke. Venous blood was collected repeatedly from 67 acute ischemic stroke patients and 15 control subjects, in order to assess NSE with ELISA, and CD45-CD34 + CD271+, CD45-CD34 + CXCR4+, CD45-CD34 + CXCR7+ and CD45-CD34 + CD133 + stem/progenitor cells by means of flow cytometry. Patients underwent repeated assessment with the National Ischemic Stroke Scale and modified Rankin Scale. Ischemic lesion volumes were assessed twice by MRI-DWI (day 1 and 5 ± 2). NSE correlated negatively with MFI levels of the CD45-CD34 + CXCR7+ cells, and percentage levels of the CD45-CD34 + and CD45-CD34 + CXCR4+ cells. NSE concentrations were significantly higher in patients compared to control subjects. NSE on day 2 positively correlated with lesion volume on both MRI. NSE on day 2 and 6-7 correlated positively with initial NIHSS scores, and on day 1 with mRS score on day 9. In conclusion, in this study NSE indicated some activity of the CD45-CD34 + CXCR7+, CD45-CD34 + and CD45-CD34 + CXCR4+ stem/progenitor cells in the first 7 days after ischemic stroke. Additionally, this study supports the thesis that NSE might be a valuable prognostic marker in acute ischemic stroke.

CD271+, CXCR7+, CXCR4+, and CD133+ Stem/Progenitor Cells and Clinical Characteristics of Acute Ischemic Stroke Patients.

Ischemic stroke causes mobilization of various groups of progenitor cells from bone marrow to bloodstream and this correlates with the neurological status of stroke patients. The goal of our study was to identify the activity of chosen progenitor/stem cells in the peripheral blood of acute ischemic stroke patients in the first 7 days after the incident, through associations between the levels of the cells and clinical features of the patients. Thirty-three acute ischemic stroke patients and 15 non-stroke control subjects had their venous blood collected repeatedly in order to assess the levels of the CD45-CD34 + CD271+, the CD45-CD34 + CXCR4+, the CD45-CD34 + CXCR7+, and the CD45-CD34 + CD133+ stem/progenitor cells by means of flow cytometry. The patients underwent repeated neurological and clinical assessments, pulse wave velocity (PWV) assessment on day 5, and MRI on day 1 and 5 ± 2. The levels of the CD45-CD34 + CXCR7+ and the CD45-CD34 + CD271+ cells were lower in the stroke patients compared with the control subjects. Only the CD45-CD34 + CD271+ cells correlated positively with lesion volume in the second MRI. The levels of the CD45-CD34 + CD133+ cells on day 2 correlated negatively with PWV and NIHSS score on day 9. The patients whose PWV was above 10 m/s had significantly higher levels of the CD45-CD34 + CXCR4+ and the CD45-CD34 + CXCR7+ cells on day 1 than those with PWV below 10 m/s. This study discovers possible activity of the CD45-CD34 + CD271+ progenitor/stem cells during the first 7 days after ischemic stroke, suggests associations of the CD45-CD34 + CD133+ cells with the neurological status of stroke patients, and some activity of the CD45-CD34 + CD133+, the CD45-CD34 + CXCR4+, and the CD45-CD34 + CXCR7+ progenitor/stem cells in the process of arterial remodeling.

Hypertension is associated with dysfunction of both peripheral and central auditory system.

OBJECTIVES: Arterial hypertension negatively influences the peripheral auditory system, causing sensorineural hearing loss. Much less is known about the detrimental effects of hypertension on the central auditory functions. METHODS: We tested 32 arterial hypertension patients and 32 age and sex-matched healthy volunteers with the expanded tonal audiometry (0.125-12.5 kHz), distortion product otoacoustic emissions (0.75-8 kHz), horizontal minimum audible angle test for eight azimuths with binaural stimulation and the random gap detection test. RESULTS: Peripheral hearing of the hypertensive patients was impaired in comparison with the controls within all audiometric frequencies (0.125-12.5 kHz) and within specific groups of frequencies. Distortion product otoacoustic emission results were significantly lower for frequencies 4 (P = 0.04) and 6 kHz (P < 0.001). The sound localization ability in the horizontal minimum audible angle test was significantly worse in the hypertensive patients in the 0°, 45°, 90°, 135°, and 270° azimuth when the interaural pure tone average (0.5-1-2 kHz) was set less than 20 dB hearing level (P < 0.05), and in the 0°, 90°, 225°, and 270°azimuth when the binaural pure tone average (0.5-1-2 kHz) was set 20 dB or less hearing level (P < 0.05). Gap detection thresholds in the random gap detection test did not differ between the two groups. CONCLUSION: Arterial hypertension is independently related to the damage of the peripheral part of the auditory system resulting in high-frequency hearing loss. Hypertensive disturbances of central auditory processing are more discrete and concern the spatial hearing resolution.

Auditory spatial deficits in brainstem disorders.

PURPOSE: Brainstem disorders seem to negatively influence the central auditory system, causing spatial hearing deficits. MATERIAL AND METHODS: We tested 11 patients with brainstem lesions due to ischemic stroke (IS), multiple sclerosis (MS), or cerebellopontine angle tumor (CPAT) together with 50 age- and sex-matched healthy volunteers. We used pure tone audiometry (PTAud), brainstem auditory evoked potentials (BAEPs) and the horizontal minimum audible angle test (HMAAT) for 8 azimuths with binaural stimulation. RESULTS: The chosen patients and the controls had normal or near normal hearing in PTAud. BAEPs interaural wave I-V latency difference was over 7 times longer in the patients group compared to the controls. Additionally, 9 of the 11 patients (81.1%) had abnormal HMAAT results. The biggest quantitative disturbances in HMAAT were present in the CPAT and the MS patients. The sound localization ability in HMAAT was significantly worse in the patients in 0° azimuth in comparison with the controls, and in 45° and 90° azimuth in patients with auditory pathway involvement compared with the ones without the involvement. CONCLUSIONS: Our study confirms the strong relationship between various brainstem pathologies and sound localization disability and sheds some light on the complexity of the relationship.

Auditory deficits in neurological disorders.

Neurological diseases present with diverse and often complex symptomatology. Focal neurological signs such as paresis, aphasia or visual field deficits together with often serious general state of a neurological patient usually push auditory symptoms into the background. Here, we present a review of literature on central and peripheral auditory disturbances that can appear in the course of most common neurological diseases. We present: cerebral stroke, cochleovestibular nerve compression syndrome, cerebral palsy, multiple sclerosis, epilepsy, myasthenia gravis and brain tumors. We focus on the neuroanatomical basis of auditory dysfunctions, their character and prevalence typical for the abovementioned diseases. Theoretical considerations are supported by broad audiological and neuroimaging studies of our patients. Auditory symptoms in neurological diseases seem to be rare. However, knowledge of these symptoms and their origin can be helpful in proper diagnosis and comprehensive patient management.

Auditory Spatial Deficits in the Early Stage of Ischemic Cerebral Stroke.

BACKGROUND: Clinical research, together with computed tomography/magnetic resonance imaging findings, proves that ischemic stroke (IS) that damages auditory pathways can cause hearing loss and impairment of higher auditory processes such as sound localization. The goal of the study was to find possible correlations between the IS risk factors, ischemic lesion volume and localization, neurologic status, and the sound localization capability in acute IS patients. METHODS: We consecutively enrolled 61 IS patients into the study. The control group consisted of 60 healthy volunteers. All neuro-otological evaluations were performed up to 30 days from the incidence of stroke. All the subjects underwent the horizontal minimum audible angle test (HMAAT) and standard tonal and speech audiometric assessments. RESULTS: HMMAT results were significantly worse in the IS patients and were present in 82.0% of the patients. There were more patients with unilateral disturbances than with bilateral ones (54.1% versus 27.9%). It was the characteristics of the ischemic lesions that correlated strongly with the sound localization deterioration, that is, their bilateral (the 90° azimuth, P = .018; the 180°, P = .002), multiple (the 45°, P = .020; the 180°, P = .007; the 225°, P = .047), and lacunar character (the 90°, P = .015; the 225°, P = .042). Differences in the types of HMAAT results were significant for lesions in the frontal and the temporal lobe (P = .018 and P = .040). In addition, worse sound localization ability was more common in patients with poor speech discrimination and the bilateral sensorineural hearing loss. We have not found statistically significant correlations for other analyzed factors such as the cortical/subcortical character of the lesions, the patients' neurologic status, and cerebrovascular risk factors. CONCLUSIONS: Sound localization impairment is common in IS patients and it is the multiple, bilateral, and lacunar character of the ischemic lesions that seems to be strongly positively correlated with the disturbance of the sound localization ability.

Hypertension and cochlear hearing loss.

This paper presents a review of experimental and clinical research on the contribution of hypertension to cochlear hearing loss. Hypertension is one of the crucial risk factors underlying pathophysiological processes taking place in the cochlea. Several mechanisms explaining these processes have been described, mainly in animal models, such as the disturbance of the inner ear potassium recycling process due to the detrimental action of natriuretic hormone, and the decrease in the cochlear oxygen partial pressure. Current evidence linking hypertension to sensorineural high-frequency cochlear hearing loss in humans may be confounded by other concomitant diseases or risk factors such as age, coronary artery disease, diabetes, obesity, hyperlipidemia, smoking and noise exposure. Therefore, further research in this field is clearly needed.

CD34/CXCR4 stem cell dynamics in acute stroke patients.

INTRODUCTION: Non-hematopoietic stem cells may be a source of paracrine and structural regeneration for brain damaged by acute ischemic stroke. In this study, we investigated correlations of CD34-, CD34/CXCR4-, and CXCR4-positive peripheral blood CD45-negative stem cells with the neurological and functional status of 34 acute stroke patients. MATERIAL AND METHODS: Blood was sampled and assessed by flow cytometry on days 1, 2, 4, and 6 after stroke onset. Parallel to blood sampling and after 3 and 6 months, patients were assessed with the National Institute of Health Stroke Scale, Barthel Index, Scandinavian Stroke Scale, and modified Rankin Scale. Blood was also sampled in 15 control subjects matched for age and sex, without history of previous stroke. RESULTS: We observed very low levels of the observed stem cells resident in peripheral blood. Higher baseline numbers of all 3 stem cell types correlated with better neurological or functional status on admission. Additionally, higher increases in CD34- and CD34/CXCR4-positive stem cell number and lower increase in CXCR4-positive cells correlated with initially worse neurological status. However, increased CD34- and CD34/CXCR4-positive cell induction in patients correlated with better functional/neurological status after the 6-month follow-up. CONCLUSIONS: We report that the number and dynamic changes in levels of non-hematopoietic CD34-, CD34/CXCR4-, and CXCR4-positive stem cells tend to correlate with acute stroke patients' neurologic and functional status.