Yasin Hamarat

Amplex Red is a widely used fluorogenic probe for hydrogen peroxide (H₂O₂) detection. In the presence of horseradish peroxidase (HRP), it reacts with H₂O₂ to form fluorescent resorufin. However, during high-voltage electric pulse treatment, fluorescence readouts may be influenced not only by H₂O₂ generation but also by the medium composition, pulse conditions, pH-dependent effects, and metal ions released as a result of electrode corrosion. In this study, we examined how medium composition and selected metal ions relevant to electroporation conditions, such as Fe²⁺, Fe³⁺, Al³⁺, and Cr⁶⁺, affect the Amplex Red/HRP/H₂O₂ fluorescence signal. Fluorescence intensity differed markedly among media, with the highest signal in phosphate-buffered saline (PBS, ~34,000 a.u.), followed by HB1 buffer (~19,000 a.u.), cell culture medium (~9000 a.u.), and distilled water (~4100 a.u.). To model ion-mediated interference, metal ions were added directly to the assay mixture. All tested ions reduced fluorescence in a concentration-dependent manner. At 0.4 µM H₂O₂, Fe²⁺ reduced fluorescence by up to 92%, Fe³⁺ by 60%, Al³⁺ by 35%, and Cr⁶⁺ by 40%. At 10 µM H₂O₂, the quenching effect was reduced but remained evident. In addition, the fluorescence readout showed strong dependence on medium composition and its pH, further demonstrating its vulnerability to the chemical environment. These findings show that the Amplex Red assay is highly sensitive to the assay environment and metal-ion interference. Therefore, it should not be used as a standalone, interference-free measure of H₂O₂ in electroporation-related experiments without appropriate validation controls.

Introduction: Postoperative delirium affects up to 60% of cardiac surgical patients. No reliable gold standard method exists for preventing delirium after cardiac surgery. An example of patient-personal monitoring is cerebral autoregulation (CA). This study aims to highlight the personal monitoring of patients’ cerebral autoregulation and to determine its relationship with postoperative delirium. Additionally, it seeks to test the hypothesis that the duration of CA impairment influences the onset of postoperative delirium. Methods: A prospective study was conducted in 2021–2023. After approval of the Ethics Committee and with the patient’s written consent, 104 adult patients undergoing elective coronary artery bypass graft (CABG) with cardiopulmonary bypass (CPB) surgery were enrolled. To diagnose possible delirium, all patients underwent a Confusion Assessment Method for the Intensive Care Unit (CAM–ICU) test. CA monitoring using transcranial Doppler was performed. CA status index – Mx was recorded. Results: Our study found that 12.5% of patients were diagnosed with delirium after on-pump CABG surgery. The total duration of cerebral autoregulation impairment (TCAI) was longer in the delirium group, 4783.0 seconds versus 4204.5 seconds ( p = .047), with a cut-off at 4380 s. Longer cardiopulmonary bypass (CPB) leads to prolonged CA impairment ( p < .001). The mean arterial pressure (MAP) during CPB was 69.67 mmHg in the non-delirium group and 74.91 mmHg in the delirium group ( p = .001), with a cutoff at 73.669 mmHg. Conclusions: CA impairment is crucial for delirium development after cardiac surgery. The duration of the TCAI event increases the risk of delirium.

Intracranial pressure (ICP) pulse wave morphology, including the ratios of the three characteristic peaks (P1, P2, and P3), offers valuable insights into intracranial dynamics and brain compliance. Traditional invasive methods for ICP pulse wave monitoring pose significant risks, highlighting the need for non-invasive alternatives. This pilot study investigates a novel non-invasive method for monitoring ICP pulse waves through closed eyelids, using a specially designed, liquid-filled, fully passive sensor system named ‘Archimedes 02’. To our knowledge, this is the first technological approach that enables the non-invasive monitoring of ICP pulse waveforms via closed eyelids. This study involved 10 healthy volunteers, aged 26–39 years, who underwent resting-state non-invasive ICP pulse wave monitoring sessions using the ‘Archimedes 02’ device while in the supine position. The recorded signals were processed to extract pulse waves and evaluate their morphological characteristics. The results indicated successful detection of pressure pulse waves, showing the expected three peaks (P1, P2, and P3) in all subjects. The calculated P2/P1 ratios were 0.762 (SD = ±0.229) for the left eye and 0.808 (SD = ±0.310) for the right eye, suggesting normal intracranial compliance across the cohort, despite variations observed in some individuals. Physiological tests—the Valsalva maneuver and the Queckenstedt test, both performed in the supine position—induced statistically significant increases in the P2/P1 and P3/P1 ratios, supporting the notion that non-invasively recorded pressure pulse waves, measured through closed eyelids, reflect intracranial volume and pressure dynamics. Additionally, a transient hypoemic/hyperemic response test performed in the upright position induced signal changes in pressure recordings from the ‘Archimedes 02’ sensor that were consistent with intact cerebral blood flow autoregulation, aligning with established physiological principles. These findings indicate that ICP pulse waves and their dynamic changes can be monitored non-invasively through closed eyelids, offering a potential method for brain monitoring in patients for whom invasive procedures are not feasible.

Background/Objectives: Glaucoma and Alzheimer’s disease (AD) are neurodegenerative conditions with vascular underpinnings. This study aimed to explore the relationship between blood pressure parameters such as mean arterial pressure (MAP), pulse pressure (PP), and cerebral perfusion pressure (CPP) and cognitive performance in patients with AD, normal-tension glaucoma (NTG), and healthy controls. We hypothesized that NTG patients, like those with mild cognitive impairment (MCI), may experience subtle cognitive changes related to vascular dysregulation. Methods: Ninety-eight participants (35 NTG, 17 AD, 46 controls) were assessed for CPP, MAP, OPP, and cognitive performance. Statistical analyses compared groups and examined correlations. Results: AD patients showed lower CPP and MAP (p < 0.001), indicating systemic vascular dysfunction, while NTG patients had higher ocular perfusion pressure (OPP) (p = 0.008), suggesting compensatory mechanisms. CPP correlated with visuospatial abilities in AD (r = 0.492, p = 0.045). MAP correlated with the Clock drawing test (CDT) scores in the NTG group (r = 0.378, p = 0.025). PP negatively correlated with cognition in AD (r = −0.527, p = 0.016 for CDT scores) and controls (r = −0.440, p = 0.002 for verbal fluency and r = −0.348, p = 0.019 for total ACE scores). Conclusions: The study highlights distinct hemodynamic profiles: systemic dysfunction in AD and localized dysregulation in NTG. These findings emphasize the role of vascular dysregulation in neurodegeneration, with implications for personalized treatment approaches targeting vascular health in neurodegenerative conditions.

Background and Objectives: Normal-tension glaucoma (NTG) is a subtype of primary open-angle glaucoma characterized by progressive optic nerve damage despite intraocular pressure (IOP) remaining within the normal range. The underlying pathophysiology of NTG remains incompletely understood, and its diagnosis is often delayed due to the lack of a definitive screening tool. This study aimed to evaluate differences in intracranial pressure pulse wave amplitude recorded from closed eyelids between NTG patients and control subjects using a novel non-invasive monitoring technology. Materials and Methods: A cross-sectional observational study was conducted, enrolling NTG patients and age-matched controls. Intracranial pressure pulse wave signals were recorded from closed eyelids using the ’Archimedes’ 02 device, which employs a highly sensitive digital pressure sensor and hydromechanical coupling for signal transmission. The amplitude of recorded intracranial pressure pulse waves was analyzed and compared between groups. Statistical analyses were performed using IBM SPSS Statistics 30.0, with significance set at p < 0.05. Results: A total of 140 participants were enrolled, including 68 NTG patients and 72 controls. After applying exclusion criteria, 63 NTG patients and 68 controls were included in the final analysis. The median intracranial pressure pulse wave amplitude was significantly higher in NTG patients (0.1326 a.u.) than in controls (0.0889 a.u.), with p = 0.01. Conclusions: These findings suggest that intracranial pressure pulse wave monitoring may serve as a potential biomarker for NTG. Further studies are needed to determine the diagnostic accuracy, sensitivity, and specificity of this technology for NTG detection.

Intracranial pressure (ICP) monitoring is crucial in the management of traumatic brain injury (TBI) and other neurological conditions. Elevated ICP or too low intracranial compliance (ICC) can compromise brain perfusion. Simultaneous monitoring of ICP and ICC is needed to optimize patient-specific brain perfusion in pathological conditions. Surrogate ICC changes can be extracted by analysis of ICP pulse wave morphology. Non-invasive, fully passive sensor and ICC changes monitoring are needed. This study introduces Archimedes, a novel, fully passive, non-invasive ICP wave monitor that utilizes mechanical pulsatile movement of the eyeball to assess ICP pulse waveforms. Preliminary findings indicate a high correlation r = [0.919; 0.96] between non-invasive and invasive ICP pulse wave morphologies, demonstrating the device’s potential for accurate ICP pulse waveform monitoring. Additionally, the monitor can discern ICC changes, providing valuable insights for TBI and normal tension glaucoma patients according to the shape of non-invasive measured ICP pulse wave. The k-nearest neighbours algorithm used in preliminary glaucoma studies yielded promising diagnostic performance, with an accuracy of 0.89, sensitivity of 0.82, specificity of 1.0 and area under curve 0.91. Ethical approvals for ongoing studies have been secured. Initial results indicate that Archimedes real-time ICC non-invasive monitor is safe, cost-effective alternative to conventional monitoring techniques.

Background and Objectives: Cardiac surgery is associated with various durations of cerebral autoregulation (CA) impairment and can significantly impact cognitive function. Cognitive functions such as memory, psychomotor speed, and attention are significantly impacted after cardiac surgery, necessitating prioritization of these areas in cognitive function tests. There is a lack of research connecting cerebral autoregulation impairment to specific cognitive function domains after cardiac surgery. This study aimed to determine if impaired cerebral autoregulation is associated with postoperative memory impairment and to test the hypothesis that the duration of this impairment affects the development of postoperative memory issues. Materials and Methods: A prospective study was conducted in 2021–2023. After approval of the Ethics Committee and with patient’s written consent, 83 adult patients undergoing elective on-pump coronary artery bypass graft (CABG) surgery were enrolled. All patients were assessed for cognitive function 1 day before surgery using the Mini-Mental state examination (MMSE-2) test as a screening tool and the Hopkins Verbal Learning Test-Revised (HVLT-R) to assess memory specifically. To diagnose possible memory impairment (IM), all patients underwent a repeat assessment of cognitive function on the 7th–10th postoperative day. Cerebral autoregulation monitoring using transcranial Doppler was performed. Cerebral autoregulation status index (Mx) was recorded using Intensive Care Brain Monitoring System software, 9.1.5.23 (Cambridge, UK). Results: According to our research, the incidence of postoperative memory impairment is 30.1%. Temporary cerebral autoregulation impairment occurs in all patients undergoing elective in-pump CABG surgery. The duration of the single longest CA impairment event in seconds (LCAI) and the LCAI dose were higher in patients with postoperative memory impairment, p = 0.006 and p < 0.007, respectively. Conclusions: Cerebral autoregulation impairment is important in developing memory loss after cardiac surgery. The duration and dose of the LCAI event are predictive of postoperative memory impairment.

Intracranial pressure measurement is frequently used for diagnosis in neurocritical care but cannot always accurately predict neurological deterioration. Intracranial compliance plays a significant role in maintaining cerebral blood flow, cerebral perfusion pressure, and intracranial pressure. This study’s objective was to investigate the feasibility of transferring external pressure into the eye orbit in a large-animal model while maintaining a clinically acceptable pressure gradient between intraorbital and external pressures. The experimental system comprised a specifically designed pressure applicator that can be placed and tightly fastened onto the eye. A pressure chamber made from thin, elastic, non-allergenic film was attached to the lower part of the applicator and placed in contact with the eyelid and surrounding tissues of piglets’ eyeballs. External pressure was increased from 0 to 20 mmHg with steps of 1 mmHg, from 20 to 30 mmHg with steps of 2 mmHg, and from 30 to 50 mmHg with steps of 5 mmHg. An invasive pressure sensor was used to measure intraorbital pressure directly. An equation was derived from measured intraorbital and external pressures (intraorbital pressure = 0.82 × external pressure + 3.12) and demonstrated that external pressure can be linearly transferred to orbit tissues with a bias (systematic error) of 3.12 mmHg. This is close to the initial intraorbital pressure within the range of pressures tested. We determined the relationship between intraorbital compliance and externally applied pressure. Our findings indicate that intraorbital compliance can be controlled across a wide range of 1.55 to 0.15 ml/mmHg. We observed that external pressure transfer into the orbit can be achieved while maintaining a clinically acceptable pressure gradient between intraorbital and external pressures.