From the lees of bokbunja wine (LBW) made from Rubus coreanus Miquel, we have identified six compounds (1: trans-4-hydroxycinnamic acid; 2: trans-4-hydroxy-3-methoxycinnamic acid; 3: 3,4-dihydroxycinnamic acid; 4: 4-hydroxy-3-methoxybenzoic acid; 5: 3,5-dimethoxy-4- hydroxybenzoic acid; and 6: 3,5-dimethoxy-4-hydroxycinnamic acid (sinapic acid)) through silica gel chromatography and UHPLC-MS. The compounds 1-6 showed strong anticoagulation and platelet aggregation inhibitory activities without hemolytic effect against human red blood cells. To date, this is the first report of the in vitro anti-thrombosis activity of sinapic acid. Our results suggest that different cinnamic and benzoic acid derivatives are closely linked to the anti-thrombosis activity of LBW, and sinapic acid could be developed as a promising anti-thrombosis agent.
The work examined the effects of Ca(2+) overload and oxidative damage on erythrocytes of river lamprey Lampetra fluvialtilis. The cells were incubated for 3h with 0.1-5µM Ca(2+) ionophore ionomycin in combination with 2.5mM Ca(2+) and 10-100µM pro-oxidant agent tert-butyl hydroperoxide (tBHP). The sensitivity of lamprey RBCs to studied compounds was evaluated by the kinetics of their death. Both toxicants induced dose- and time dependent phosphatidylserine (PS) externalization (annexin V-FITC labeling) and loss of membrane integrity (propidium iodide uptake). Highest doses of ionomycin (1-2µM) increased the number of PS-exposed erythrocytes to 7-9% within 3h, while 100µM tBHP produced up to 50% of annexin V-FITC-positive cells. Caspase inhibitor Boc-D-FMK (50µM), calpain inhibitor PD150606 (10µM) and broad protease inhibitor leupeptin (200µM) did not prevent ionomycin-induced PS externalization, whereas tBHP-triggered apoptosis was blunted by Boc-D-FMK. tBHP-dependent death of lamprey erythrocytes was accompanied by the decrease in relative cell size, loss of cell viability, activation of caspases 9 and 3/7, and loss of mitochondrial membrane potential, but all these processes were partially attenuated by Boc-D-FMK. None of examined death-associated events were observed in ionomycin-treated erythrocytes except activation of caspase-9. Incubation with ionomycin did not alter intracellular K(+) and Na(+) content, while exposure to tBHP resulted in 80% loss of K(+) and 2.8-fold accumulation of Na(+). Thus, lamprey erythrocytes appear to be more susceptible to oxidative damage. Ca(2+) overload does not activate the cytosolic death pathways in these cells.
Division of Hematopathology, Department of Laboratory Medicine and Pathology, Dalhousie University, Halifax, Nova Scotia, Canada. calvino.cheng@cdha.nshealth.ca
Recent blood shortages and the potential clinical impact of red blood cell (RBC) age highlight the need to understand blood supply delivery. This study addresses the characteristics and mechanics of RBC unit trafficking and storage across the transfusion service, previously undescribed in the literature.
This retrospective qualitative institutionwide survey assessed the comprehensive RBC life cycle within Capital District Health Authority in Nova Scotia, Canada, during 2007.
A total of 15,930 unique RBC units were received from the supplier with 98.6% having mean age of 12.5 days. The mean ages on receipt, transfusion, and time spent in the transfusion system before transfusion were 12, 19.6, and 7.5 days, respectively. Of 12,298 units, 9689 (78.8%) remained within the blood transfusion services (BTSs), while 2609 (21.2%) migrated having been returned a mean of 1.26 times from locations outside BTS (SD, 0.56 times), the latter spending a mean of 18.4 hours outside the BTS. Stationary units had mean age at dispense and time spent in the transfusion system of 19.2 and 6.9 days, respectively, compared to migratory units at 21.3 and 9.9 days, respectively (p
Antifreeze proteins (AFPs) have important functions in many freeze-tolerant organisms. The proteins non-colligatively lower the freezing point and functionally inhibit ice recrystallization in frozen solutions. In our previous studies, we found that the Arctic yeast Leucosporidium sp. produces an AFP (LeIBP), and that the protein could be successfully produced in Pichia expression system. The present study showed that recombinant LeIBP possesses the ability to reduce the damage induced to red blood cells (RBCs) by freeze thawing. In addition to 40 % glycerol, both 0.4 and 0.8 mg/ml LeIBPs significantly reduced freeze-thaw-induced hemolysis at either rapid- (45 °C) or slow-warming (22 °C) temperatures. Post-thaw cell counts of the cryopreserved RBCs were dramatically enhanced, in particular, in 0.8 mg/ml LeIBP. Interestingly, the cryopreserved cells in the presence of LeIBP showed preserved cell size distribution. These results indicate that the ability of LeIBP to inhibit ice recrystallization helps the RBCs avoid critically damaging electrolyte concentrations, which are known as solution effects. Considering all these data, LeIBP can be thought of as a key component in improving RBC cryopreservation efficiency.
[Effect of diet enriched with essential nutrients and S-adenosylmethionine on the structural and functional state of erythrocytes in patients with metabolic syndrome]
An analysis of structural and functional state of erythrocytes of 136 patients with metabolic syndrome and nourishment of these patients have been outlined in the article. It was determined that the diet with decreased level of carbohydrate and enriched in amino-acids, lecithin, vitamins and antioxidants enables weight correction and normalization of structural and functional state of erythrocytes.
There has been interest in determining whether older red blood cell (RBC) units have negative clinical effects. Numerous observational studies have shown that older RBC units are an independent factor for patient mortality. However, recently published randomized clinical trials have shown no difference of clinical outcome for patients receiving old or fresh RBCs. An overlooked but essential issue in assessing RBC unit quality and ultimately designing the necessary clinical trials is a metric for what constitutes an old or fresh RBC unit.
Twenty RBC units were profiled using quantitative metabolomics over 42 days of storage in SAGM with 3- to 4-day time intervals. Metabolic pathway usage during storage was assessed using systems biology methods. The detected time intervals of the metabolic states were compared to clinical outcomes.
Using multivariate statistics, we identified a nonlinear decay process exhibiting three distinct metabolic states (Days 0-10, 10-17, and 17-42). Hematologic variables traditionally measured in the transfusion setting (e.g., pH, hemolysis, RBC indices) did not distinguish these three states. Systemic changes in pathway usage occurred between the three states, with key pathways changing in both magnitude and direction. Finally, an association was found between the time periods of the metabolic states with the clinical outcomes of more than 280,000 patients in the country of Denmark transfused over the past 15 years and endothelial damage markers in healthy volunteers undergoing autologous transfusions.
The state of RBC metabolism may be a better indicator of cellular quality than traditional hematologic variables.
Methylene di-phenyl diisocyanate (MDI) is used to make polyurethane products. The predominant occupational disease attributed to diisocyanates, including MDI, is asthma; however, the potential for genotoxicity has also been of concern. Diisocyanates are very reactive compounds that can undergo nonenzymatic hydrolysis to form methylenedianiline (MDA), or react under physiological conditions with primary amines to form ureas and/or with thiols to form labile thiol acid esters. MDA is a carcinogen in animals and a suspected carcinogen in humans. Brown Norway rats (BNR) were exposed to either 7 or 113 mg/m(3) MDI aerosol for 1 h/week x3 weeks and sacrificed 1 week later. Micronuclei (MN) formation was assessed from bone marrow polychromatic erythrocytes (PCE). A dose-dependent increase in the frequency of micronucleated polychromatic erythrocytes (MN-PCEs) was noted. In vitro exposure of Chinese hamster lung fibroblasts (V79) to MDA or MDI-thiol conjugates, but not to MDI, significantly increased the frequency of MN. MDI-thiol conjugate-exposed cell cultures did not have detectable levels of MDA. A significant increase in the number of V79 cells in metaphase, as well as the number of cells with precipitants within both the cytoplasm and nuclei, were noted in MDI-glutathione-exposed cultures. The results of this study indicate that MDI aerosol exposure can cause MN formation through either the hydrolysis of MDI to MDA or possibly the formation of thiol conjugates.
We analysed the erythrocytes and lymphocytes with micronuclei of 3902 individuals living in 16 settlements in the west of the Altai region. It was found that the majority of individuals with high frequencies of micronucleated cells were in the settlements located near to the Semipalatinsk atomic proving ground. In particular, considerable cytogenetic alterations were found in individuals born during the period of intensive testing on the Semipalatinsk proving ground (from 1949 to 1962). The data we have obtained allow us to conclude that radiation damage of cytogenetic structures in the prenatal (predelivery) and early postnatal periods leading to the formation of micronuclei can remain in the human organism for decades and, perhaps, throughout life.
Replicated adaptive radiation events, typified by phenotypic divergence across resource axes, provide important insight into the eco-evolutionary dynamics that lead to the formation of new species. Here, we show that in trimorphic adaptive radiations of European whitefish (Coregonus lavaretus), divergence of the oxygen transport system has occurred across the pelagic/littoral (shallow)-profundal (deep) resource axis, and at multiple biological scales. Profundal whitefish exhibited significantly larger red blood cells (RBCs), a greater proportion of cathodic hemoglobin protein components, and higher hemoglobin transcript abundance in kidney compared to littoral and pelagic morphs. Hemoglobin transcript abundance in brain and gill, but not kidney, and anodic hemoglobin protein component diversity in blood were also linked to variation at an intronic single nucleotide polymorphism (SNP). As the whitefish morphs differ in population genetic structure at this SNP, hemoglobin transcript and protein divergence between profundal and pelagic/littoral morphs is likely being driven by genetic divergence. Our findings, along with our previous work on lake whitefish, highlight the importance of the oxygen transport system to the postglacial colonization of novel lacustrine environments by whitefish throughout the northern hemisphere.
