Post-occlusive reactive hyperemia variables can be used to diagnose vascular dysfunction in hemorrhagic shock.

INTRODUCTION: Laser doppler flowmetry (LDF) allows non-invasive assessment of microvascular functions. The combination of LDF with an occlusion functional test enables study of post-occlusive reactive hyperemia (PORH), providing additional information about vasomotor function, capillary blood flow reserve, and the overall reactivity of the microvascular system. AIM: To identify early alterations of PORH variables in the skin of a rat in hemorrhagic shock (HS). MATERIAL AND METHODS: Male Wistar rats (n = 14) weighing 400-450 g were anesthetized with a combination of tiletamine/zolazepam (20 mg/kg) and xylazine (5 mg/kg). The animals breathed on their own, and were placed on a heated platform in the supine position. A PE-50 catheter was inserted into the carotid artery to measure the mean arterial pressure (MAP). The optical probe of the Laser Doppler device was installed on the plantar surface of the hind limb of a rat; a pneumatic cuff was applied proximal to the same limb. The occlusion time was 3 min. The following physiological variables were measured at baseline and 30 min after blood loss: MAP, mmHg; mean cutaneous blood flow (M, PU); cutaneous vascular conductance (CVC = M/MAP); peak hyperemia (Mmax, PU) and maximum cutaneous vascular conductance (CVCmax) during PORH. In the HS group (n = 7), 30 % of the estimated blood volume was taken within 5 min. There was no blood loss in the group of sham-operated animals (Sham, n = 7). The results are presented as Me [25 %;75 %]. The U-Mann-Whitney criterion was used to evaluate intergroup differences. Differences were considered statistically significant at p < 0.05. RESULTS: The groups did not differ at baseline. Blood loss led to a significant decrease in MAP (43 [31;46] vs. 94 [84;104] mmHg), M (11.5 [16.9;7.8] vs 16.7 [20.2;13.9]) and Mmax (18.1 [16.4;21.8] vs. 25.0 [23.0;26.2]) in the HS group compared to the Sham group, respectively. At the same time, both CVC (0.25 [0.23;0.30] vs. 0.16 [0.14;0.21]) and CVCmax (0.55 [0.38;0.49] vs 0.24 [0.23; 0.29]) increased after blood loss in the HS group compared to the Sham group. Arterial blood gas analysis revealed metabolic lactic acidosis in the HS group. CONCLUSION: In this rat model of HS, alterations in cutaneous blood flow are manifested by a decrease in perfusion (M) and the intensity of PORH (Mmax) with a simultaneous increase in vascular conductance (CVC and CVCmax).

A systematic evaluation of cutaneous microcirculation in the foot using post-occlusive reactive hyperemia.

OBJECTIVES: Cutaneous microcirculatory impairments are associated with skin injury to the foot. Post-Occlusive reactive hyperemia (PORH) is one of the quick and easy methods to assess microcirculatory function. However, there are variations in the protocols currently used. Hence, this study aimed to systematically investigate the reproducibility of PORH protocols with minimal occlusion time in the foot. METHODS: Post-Occlusive reactive hyperemia was measured using 12 different protocols (three occlusion times, two occlusion sites and with or without temperature control) in 25 healthy adults. Each of the 12 different protocols was repeated three times, and the intraclass correlation coefficient (ICC) was calculated. RESULTS: Intraclass correlation coefficient showed that that ankle level occlusion produced moderate to excellent reproducibility for most PORH measures. In the right foot, 30- and 60-s ankle level occlusion without temperature control showed ICC of >0.40 for all parameters except the area of hyperemia (ICC = -0.36) and biological zero to peak flow percent change (ICC = -0.46). In the left foot, 30-s ankle level occlusion without temperature control showed ICC of >0.40 for all parameters except time to latency (ICC = 0.29), after hyperemia (ICC = 0.37), and max (ICC = -0.01), and area of hyperemia (ICC = -0.36). But the 60-s protocol showed ICC > 0.40 for all except time to max (ICC = 0.38). In the hallux protocols, all three 10-, 30-, and 60-s protocols without temperature control showed moderate to excellent reproducibility (ICC > 0.40). In most cases, the temporal and area under the perfusion-time curve parameters showed poor reproducibility. CONCLUSION: Post-Occlusive reactive hyperemia can be tested efficiently with a minimal occlusion time of 10 s with hallux occlusion and 30 s with ankle occlusion in the foot. This can suggest that microcirculatory assessment is feasible in routine practice and can potentially be included for routine assessment of foot in people with diabetes.

Cord factor (trehalose 6,6'-dimycolate) forms fully stable and non-permeable lipid bilayers required for a functional outer membrane.

Cord factor (trehalose 6,6'-dimycolate, TDM) is the major lipid in the outer membrane of Corynebacteria and Mycobacteria. Although its role is well recognized in the immune response phenomena, its membrane biophysical properties remained largely unexplored and TDM has often been described as a detergent. We purified the main components of the outer membrane from Corynebacterium glutamicum and analyzed their membrane forming properties. In mixture with endogenous cardiolipin, but not alone, the spontaneous hydration of TDM produces liposomes. As a pure component, TDM formed vesicles only by the detergent dialysis method. Perdeuterated cardiolipin-TDM mixtures were shown by deuterium nuclear magnetic resonance (NMR) to exhibit a gel to liquid crystalline phase transition over a 273-295K temperature range, for cells grown at 303K, and thus to be in a liquid crystalline state at physiological temperature. Molecular dynamics simulations of hydrated TDM bilayers provided the trehalose average orientation and conformation, the chain order parameters, the area per lipid and the bilayer thickness which was confirmed by electron microscopy. Finally the Porin A-Porin H ion channel from the Corynebacterial outer membrane was reconstituted in TDM liposomes. With properly mycoloylated proteins, it manifested the typical voltage dependent ion channel properties of an outer membrane porin.