An Exercise Immune Fitness Test to Unravel Disease Mechanisms-A Proof-of-Concept Heart Failure Study.

Background: Cardiorespiratory fitness positively correlates with longevity and immune health. Regular exercise may provide health benefits by reducing systemic inflammation. In chronic disease conditions, such as chronic heart failure and chronic fatigue syndrome, mechanistic links have been postulated between inflammation, muscle weakness, frailty, catabolic/anabolic imbalance, and aberrant chronic activation of immunity with monocyte upregulation. We hypothesize that (1) temporal changes in transcriptome profiles of peripheral blood mononuclear cells during strenuous acute bouts of exercise using cardiopulmonary exercise testing are present in adult subjects, (2) these temporal dynamic changes are different between healthy persons and heart failure patients and correlate with clinical exercise-parameters and (3) they portend prognostic information. Methods: In total, 16 Heart Failure (HF) patients and 4 healthy volunteers (HV) were included in our proof-of-concept study. All participants underwent upright bicycle cardiopulmonary exercise testing. Blood samples were collected at three time points (TP) (TP1: 30 min before, TP2: peak exercise, TP3: 1 h after peak exercise). We divided 20 participants into 3 clinically relevant groups of cardiorespiratory fitness, defined by peak VO2: HV (n = 4, VO2 ≥ 22 mL/kg/min), mild HF (HF1) (n = 7, 14 < VO2 < 22 mL/kg/min), and severe HF (HF2) (n = 9, VO2 ≤ 14 mL/kg/min). Results: Based on the statistical analysis with 20-100% restriction, FDR correction (p-value 0.05) and 2.0-fold change across the three time points (TP1, TP2, TP3) criteria, we obtained 11 differentially expressed genes (DEG). Out of these 11 genes, the median Gene Expression Profile value decreased from TP1 to TP2 in 10 genes. The only gene that did not follow this pattern was CCDC181. By performing 1-way ANOVA, we identified 8/11 genes in each of the two groups (HV versus HF) while 5 of the genes (TTC34, TMEM119, C19orf33, ID1, TKTL2) overlapped between the two groups. We found 265 genes which are differentially expressed between those who survived and those who died. Conclusions: From our proof-of-concept heart failure study, we conclude that gene expression correlates with VO2 peak in both healthy individuals and HF patients, potentially by regulating various physiological processes involved in oxygen uptake and utilization during exercise. Multi-omics profiling may help identify novel biomarkers for assessing exercise capacity and prognosis in HF patients, as well as potential targets for therapeutic intervention to improve VO2 peak and quality of life. We anticipate that our results will provide a novel metric for classifying immune health.

Real-Time Characterization of Cell Membrane Disruption by α-Synuclein Oligomers in Live SH-SY5Y Neuroblastoma Cells.

Aggregation of the natively unfolded protein α-synuclein (α-Syn) has been widely correlated to the neuronal death associated with Parkinson's disease. Mutations and protein overaccumulation can promote the aggregation of α-Syn into oligomers and fibrils. Recent work has suggested that α-Syn oligomers can permeabilize the neuronal membrane, promoting calcium influx and cell death. However, the mechanism of this permeabilization is still uncertain and has yet to be characterized in live cells. This work uses scanning ion conductance microscopy (SICM) to image, in real time and without using chemical probes, the topographies of live SH-SY5Y neuroblastoma cells after exposure to α-Syn oligomers. Substantial morphological changes were observed, with micrometer-scale hills and troughs observed at lower α-Syn concentrations (1.00 µM) and large, transient pores observed at higher α-Syn concentrations (6.0 µM). These findings suggest that α-Syn oligomers may permeabilize the neuronal membrane by destabilizing the lipid bilayer and opening transient pores.

Real-time determination of aggregated alpha-synuclein induced membrane disruption at neuroblastoma cells using scanning ion conductance microscopy.

Parkinson's disease (PD) is recognized as the second most common neurodegenerative disorder and has affected approximately one million people in the United States alone. A large body of evidence has suggested that deposition of aggregated alpha-synuclein (α-Syn), a brain protein abundant near presynaptic termini, in intracellular protein inclusions (Lewy bodies) results in neuronal cell damage and ultimately contributes to the progression of PD. However, the exact mechanism is still unclear. One hypothesis is that α-Syn aggregates disrupt the cell membrane's integrity, eventually leading to cell death. We used scanning ion conductance microscopy (SICM) to monitor the morphological changes of SH-SY5Y neuroblastoma cells and observed dramatic disruption of the cell membrane after adding α-Syn aggregates to the culturing media. This work demonstrates that SICM can be applied as a new approach to studying the cytotoxicity of α-Syn aggregates.

Clinical performance of KeraSoft(®) IC in irregular corneas.

PURPOSE: This study evaluated the clinical performance of KeraSoft(®) IC (KIC) soft contact lenses in subjects with irregular corneas. PATIENTS AND METHODS: This was a 12-month, prospective, open-label, observational study, which enrolled 43 subjects who were 18 years of age or older with irregular corneas. Subjects were fit according to the KIC Fitting Manual (kerasoftic.com). After achieving best fit according to the fitting manual, lenses were assessed for comfort, vision, centration, rotation, and movement. Subjects were instructed to wear their lenses between 8 and 16 hours each day. Assessments at the exit visit included logMAR visual acuity with high and low contrast, spherocylindrical overrefraction, slit-lamp findings, adverse events, and subjective outcomes. RESULTS: The average base curve was 8.17±0.32 mm (n=70 eyes), and the average diameter dispensed was 14.53±0.12 mm (n=70 eyes). From the baseline to 12 months, there was statistically significant improvement in logMAR visual acuity with high contrast (P=0.038), but no significant difference in low-contrast visual acuity was observed (P>0.05). Slit-lamp findings were ≤ grade 1 for the majority of subjects (89%). Two nonserious adverse events were reported for two of the 84 enrolled eyes (two subjects). At 12 months, subjects reported improvements from habitual baseline for comfort and vision, both upon insertion and just before removal of lenses. CONCLUSION: Clinical outcomes at 12 months showed good visual, safety, and subjective outcomes for subjects with corneal irregularities who wore KeraSoft(®) IC soft contact lenses.

Correlation of tear inflammatory cytokines and matrix metalloproteinases with four dry eye diagnostic tests.

PURPOSE: Tear cytokines and matrix metalloproteinases (MMPs) can be extracted from the Schirmer strip. This study examined the extracted levels of tear cytokines and MMPs from Schirmer strips and potential correlation with Schirmer's test, tear breakup time (TBUT), tear osmolarity, and ocular surface disease index (OSDI). METHODS: Thirty healthy volunteers were clinically evaluated for known methods to diagnose dry eye disease, including Schirmer's test, tear osmolarity, OSDI, and TBUT. Tears were collected by Schirmer strips and proteins were extracted from the Schirmer strip in 0.5 M NaCl with 0.5% Tween 20 and analyzed using multiplex assay kits to examine cytokines or MMPs. Calculated cytokine and MMP concentrations for all samples were sorted into groups according to a positive or negative for each of the above-cited four dry eye diagnostic tests, individually and in combination. RESULTS: Five inflammatory cytokines (IL-1α, -1ß, -6, -8, and TNF-α) and five MMPs (MMPs 1, 2, 7, 9, and 10) were extracted from clinical Schirmer strips. Schirmer strip measurement and tear osmolarity correlated well with increased concentrations of the inflammatory cytokines and MMPs, whereas TBUT and OSDI did not. CONCLUSIONS: Both the Schirmer's test and tear osmolarity may be more relevant to the clinician in the diagnosis of ocular surface diseases with an increased level of inflammatory mediators.

A method to extract cytokines and matrix metalloproteinases from Schirmer strips and analyze using Luminex.

PURPOSE: The Schirmer's test is commonly used in the clinic for the diagnosis of dry eye disease by measuring tear volume. This report describes a procedure which can be used to recover tears from the Schirmer strip for the measurement of multiple tear cytokines as well as matrix metalloproteinases (MMPs) by Luminex technology. METHODS: Cytokine and MMP recovery was determined by using spiked Schirmer strips presoaked with known cytokines or MMPs prepared in PBS with 1% BSA. In a clinical study, tears were collected from 5 subjects using Schirmer strips. Strips were stored on ice immediately after removal from the subject and stored dry at -20 °C for 16-24 h. Cytokines were extracted from the Schirmer strip in 0.5 M NaCl with 0.5% Tween-20. Concentrations of cytokines and MMPs in collected tear samples were analyzed by Luminex using both a 10-cytokine and a 5-MMP kit. RESULTS: The standard curves for the assay in both the kit assay buffer and extraction buffer were identical for 9 of the 10 cytokines and all 5 MMPs. In the clinical sample all the cytokines (interleukin 1α [IL-1α], IL-1ß, IL-1ra, IL-4, IL-6, IL-8, IL-10, IL-13, monocyte chemotactic protein-1 [MCP-1], and tumor necrosis factor-α [TNF-α]) and 5 MMPs (MMP-1, MMP-2, MMP-7, MMP-9, and MMP-10) tested were detected in at least 50% of the 10 subject samples. Recoveries from extracted Schirmer strips were >60% for 8 of the 10 cytokines and all MMPs. CONCLUSIONS: Numerous cytokines and MMPs were detected in the tear samples collected using the Schirmer strip, including many that have been implicated in ocular surface disease. This procedure may be used to evaluate the cytokine and MMP content in tear samples in clinical studies, especially for the evaluation of dry eye therapeutics. Because the Schirmer test is routine in the assessment of dry eye, this method offers the opportunity to evaluate both the quantity and quality of the tears.

A comparison of polysomnography and a portable home sleep study in the diagnosis of obstructive sleep apnea syndrome.

OBJECTIVE: To validate the role of a portable sleep monitor device (SNAP) in the diagnosis of obstructive sleep apnea syndrome (OSAS). Inter-reader variability was also assessed for both PSG and SNAP. STUDY DESIGN AND SETTING: Sixty consecutive adults referred for PSG at The University of Chicago Sleep Disorder Clinic were prospectively enrolled. RESULTS: There was no significant difference between total number of apnea and hypopnea, respiratory disturbance index (RDI), and minimum oxygen obtained by PSG and SNAP, but there was a significant difference between sleep time and mean oxygen. Pearson's correlation coefficient for RDI > or = 15 was 0.92. CONCLUSION: There was a significant correlation of RDIs between SNAP and PSG. SNAP has good sensitivity, specificity, positive and negative predictive values. Differences between SNAP and PSG could be attributed to inter-reader variability and not necessarily due to technical limitations of SNAP. SNAP is an excellent tool for the diagnosis of OSAS in the laboratory setting. Future studies should be performed to evaluate SNAP's accuracy in the home setting in the diagnosis of OSAS. EBM RATING: B-2.