Chain-Length Dependence of Peptide-Lipid Bilayer Interaction Strength and Binding Kinetics: A Combined Theoretical and Experimental Approach.

Physical interactions between polypeptide chains and lipid membranes underlie critical cellular processes. Yet, despite fundamental importance, key mechanistic aspects of these interactions remain elusive. Bulk experiments have revealed a linear relationship between free energy and peptide chain length in a model system, but does this linearity extend to the interaction strength and to the kinetics of lipid binding? To address these questions, we utilized a combination of coarse-grained molecular dynamics (CG MD) simulations, analytical modeling, and atomic force microscopy (AFM)-based single molecule force spectroscopy. Following previous bulk experiments, we focused on interactions between short hydrophobic peptides (WLn, n = 1, ..., 5) with 1-palmitoyl-2-oleoyl-glycero-3-phosphocholine (POPC) bilayers, a simple system that probes peptide primary structure effects. Potentials of mean force extracted from CG MD recapitulated the linearity of free energy with the chain length. Simulation results were quantitatively connected to bulk biochemical experiments via a single scaling factor of order unity, corroborating the methodology. Additionally, CG MD revealed an increase in the distance to the transition state, a result that weakens the dependence of the dissociation force on the peptide chain length. AFM experiments elucidated rupture force distributions and, through modeling, intrinsic dissociation rates. Taken together, the analysis indicates a rupture force plateau in the WLn-POPC system, suggesting that the final rupture event involves the last 2 or 3 residues. In contrast, the linear dependence on chain length was preserved in the intrinsic dissociation rate. This study advances the understanding of peptide-lipid interactions and provides potentially useful insights for the design of peptides with tailored membrane-interacting properties.

Use of near infrared spectroscopy to detect impaired tissue oxygen saturation in patients with complex regional pain syndrome type 1.

PURPOSE: Deep tissue hypoxia has been hypothesized in the pathogenesis of complex regional pain syndrome type 1 (CRPS 1) for some patients. The purpose of this study was to determine if near-infrared spectroscopy (NIRS) could detect differences in deep tissue oxygen saturation (StO2) and microcirculatory function in the hands of patients with CRPS 1. METHODS: Tissue oxygen saturation was evaluated at baseline and during an ischemia reperfusion challenge using vascular occlusion testing (VOT) in affected vs unaffected hands of patients with unilateral upper limb CRPS 1. A non-randomized experimental study design was used with baseline StO2 as the primary outcome measure. Secondary outcome measures were occlusion and reperfusion slopes from VOT. Values were compared with the unaffected, contralateral hand and with the dominant and non-dominant hands of sex and age-matched volunteers. Correlations between values derived from NIRS and measures of pain and function from the Brief Pain Inventory (BPI) and the Disability of the Arm, Shoulder and Hand (DASH) questionnaires were explored. RESULTS: Independent of handedness, the baseline StO2 of the affected hands of ten CRPS 1 patients was significantly lower than that of their unaffected hands (-5.8%; 95% confidence interval [CI] -10.6 to -1.0; P = 0.02). The baseline StO2 of affected CRPS 1 hands was also significantly lower than the non-dominant hands of ten volunteers (-7.3%; 95% CI -12.4 to -2.3; P = 0.007). Differences in VOT occlusion and reperfusion slopes did not reveal changes that could be uniquely attributed to CRPS 1. No significant correlations were detected between values derived from VOT and values for pain and function obtained from BPI and DASH questionnaires for patients with CRPS 1. CONCLUSIONS: Hands of patients affected by CRPS 1 of the upper limb showed significantly lower StO2 compared with their unaffected contralateral hand as well as the hands of control subjects. This trial was registered at: ClinicalTrials.gov: NCT01586377.

A novel assessment of peripheral tissue microcirculatory vasoreactivity using vascular occlusion testing during cardiopulmonary bypass.

OBJECTIVE: Cardiac surgery and cardiopulmonary bypass are associated with release of inflammatory mediators and microcirculatory alterations that result in organ dysfunction. Near-infrared spectroscopic measurement of tissue oxygen saturation (StO2) and the vascular occlusion test (VOT) were utilized in a study of elective cardiac surgical patients as a novel, noninvasive method of assessing microcirculatory vasoreactivity during nonpulsatile cardiopulmonary bypass (CPB). The objective of this pilot study was to determine whether differences in microcirculatory function and vasoreactivity could be measured in cardiac surgery using StO2 and VOT. DESIGN: A prospective, observational trial. SETTING: Tertiary care teaching hospital. PARTICIPANTS: Thirteen patients undergoing elective cardiac surgery using tepid, nonpulsatile cardiopulmonary bypass. INTERVENTIONS: Patients had continuous regional oxygen saturation monitoring using near-infrared spectroscopy and vascular occlusion tests performed in the perioperative period before, during, and after cardiopulmonary bypass. MEASUREMENTS AND MAIN RESULTS: Occlusion slope and hyperemic area did not vary significantly. Mean reperfusion slope was significantly lower during cardiopulmonary bypass (2.4%/second) compared to before and after bypass (4.1 and 3.5%/second, respectively). Reperfusion slope decreased as a function of CPB duration. CONCLUSIONS: This pilot study demonstrates a significant difference in reperfusion slopes during cardiopulmonary bypass when compared to prebypass and postbypass, suggesting impaired peripheral microvascular reactivity. Reperfusion slopes also exhibited a successive decline with duration of CPB, implying worsening microcirculatory dysfunction that returned to baseline values in all patients within 1 hour of separation from CPB. This noninvasive technique has potential to optimize circulatory parameters during cardiopulmonary bypass.

Severe hypokalemia, paralysis, and AIDS-associated isospora belli diarrhea.

BACKGROUND: Hypokalemia of clinical significance, severe enough to cause paralysis and cardiac dysrhythmias, is an uncommon life-threatening medical disorder. Hypokalemic periodic paralysis (HPP), where an abrupt intracellular shift of potassium has occurred, must be distinguished from hypokalemic non-periodic paralysis (non-HPP), where a total body potassium deficit exists. The challenge for emergency physicians is to swiftly and accurately differentiate disease etiology and institute prompt treatment. Aggressive potassium repletion, required in non-HPP etiologies, may predispose HPP patients to dangerous and potentially fatal rebound hyperkalemia as the intracellular potassium shift resolves. OBJECTIVES: Describe the process by which HPP and non-HPP can be differentiated on clinical and laboratory grounds leading to appropriate resuscitation from severe and clinically relevant hypokalemia. Chronic diarrhea syndromes and possibly normal alkaline phosphatase levels alert the clinician to the potential for non-HPP. CASE REPORT: A case of HIV/AIDS-associated isospora belli diarrheal illness with severe hypokalemia of the non-HPP type is presented. Historical, laboratory, and clinical findings, which assisted in the differentiation from HPP, are emphasized. Clinical progression and treatment strategies, as well as electrocardiogram findings with T-U-P fusion waves, are described in detail. CONCLUSION: The history, especially the time course of disease, is essential in differentiating HPP and non-HPP, allowing treatment to proceed without overcorrection and rebound hyperkalemia. Although other infectious diseases have been linked to non-HPP, to our knowledge this is the first report linking non-HPP to isospora belli diarrheal illness.

Diffusion Tensor Imaging (DTI) Correlates of Self-Reported Sleep Quality and Depression Following Mild Traumatic Brain Injury.

Background: Mild traumatic brain injuries (mTBIs) are a significant social, sport, and military health issue. In spite of advances in the clinical management of these injuries, the underlying pathophysiology is not well-understood. There is a critical need to advance objective biomarkers, allowing the identification and tracking of the long-term evolution of changes resulting from mTBI. Diffusion-weighted imaging (DWI) allows for the assessment of white-matter properties in the brain and shows promise as a suitable biomarker of mTBI pathophysiology. Methods: 34 individuals within a year of an mTBI (age: 24.4 ± 7.4) and 18 individuals with no history of mTBI (age: 23.2 ± 3.4) participated in this study. Participants completed self-report measures related to functional outcomes, psychological health, post-injury symptoms, and sleep, and underwent a neuroimaging session that included DWI. Whole-brain white matter was skeletonized using tract-based spatial statistics (TBSS) and compared between groups as well as correlated within-group with the self-report measures. Results: There were no statistically significant anatomical differences between the two groups. After controlling for time since injury, fractional anisotropy (FA) demonstrated a negative correlation with sleep quality scores (higher FA was associated with better sleep quality) and increasing depressive symptoms in the mTBI participants. Conversely, mean (MD) and radial diffusivity (RD) demonstrated positive correlations with sleep quality scores (higher RD was associated with worse sleep quality) and increasing depressive symptoms. These correlations were observed bilaterally in the internal capsule (anterior and posterior limbs), corona radiata (anterior and superior), fornix, and superior fronto-occipital fasciculi. Conclusion: The results of this study indicate that the clinical presentation of mTBI, particularly with respect to depression and sleep, is associated with reduced white-matter integrity in multiple areas of the brain, even after controlling for time since injury. These areas are generally associated not only with sleep and emotion regulation but also cognition. Consequently, the onset of depression and sleep dysfunction as well as cognitive impairments following mTBI may be closely related to each other and to white-matter integrity throughout the brain.