Cold stored platelets in the management of bleeding: is it about bioenergetics?

When platelet concentrates (PCs) were first introduced in the 1960s as a blood component therapy, they were stored in the cold. As platelet transfusion became more important for the treatment of chemotherapy-induced thrombocytopenia, research into ways to increase supply intensified. During the late 1960s/early 1970s, it was demonstrated through radioactive labeling of platelets that room temperature platelets (RTP) had superior post-transfusion recovery and survival compared with cold-stored platelets (CSP). This led to a universal switch to room temperature storage, despite CSP demonstrating superior hemostatic effectiveness upon being transfused. There has been a global resurgence in studies into CSP over the last two decades, with an increase in the use of PC to treat acute bleeding within hospital and pre-hospital care. CSP demonstrate many benefits over RTP, including longer shelf life, decreased bacterial risk and easier logistics for transport, making PC accessible in areas where they have not previously been, such as the battlefield. In addition, CSP are reported to have greater hemostatic function than RTP and are thus potentially better for the treatment of bleeding. This review describes the history of CSP, the functional and metabolic assays used to assess the platelet storage lesion in PC and the current research, benefits and limitations of CSP. We also discuss whether the application of new technology for studying mitochondrial and glycolytic function in PC could provide enhanced understanding of platelet metabolism during storage and thus contribute to the continued improvements in the manufacturing and storage of PC.

What is the context? To transition into an activated state, platelets require a highly efficient source of energy that is met through the production of ATP ­ this is referred to as "platelet bioenergetics"Platelets can be removed from healthy donors and used to make platelet concentrates for clinical usePlatelet concentrates are used clinically either therapeutically (to halt bleeding) or prophylactically (to prevent bleeding in patients with low platelet counts)They are stored at room temperature (20­24^oC) with constant gentle agitation, in packs that allow gas exchange and have a 7-day shelf life in some jurisdictionsStoring platelets in the cold (2­6^oC) has historically been shown to improve their ability to halt bleedingWhat is new? There is a renewed interest in cold stored platelets for use in actively bleeding patientsThere are benefits to cold-storing platelets over room temperature storageCold stored platelets are licensed in the US and Norway for certain indications for 14 daysWhat is next? Cold stored platelets have the potential to improve logistics of clinical supply of platelets, enable supply of platelet concentrates where access is currently limited, such as pre-hospital care and on the battlefield and provide improved hemostatic effects for bleeding patients.New research measuring the bioenergetic profiles of cold stored platelets could advance understanding of metabolism in cold stored platelets and support decisions on their re-introduction on a wider scale.

Implementation of Standardized Care for the Medical Stabilization of Patients With Anorexia Nervosa.

Approaches to refeeding patients with anorexia nervosa for medical stabilization vary across institutions, and there is no established standard of care. This study assessed the impact of a refeeding pathway on hospital length of stay and transfer to the psychiatry unit. Methods: This quality improvement intervention sought to standardize care for adolescents with anorexia nervosa at a tertiary care, free-standing children's hospital from Spring 2017 to Fall 2018. The pathway specified admission criteria, nutritional advancement, activity restriction, laboratory monitoring, readiness to transfer to the psychiatry unit, and discharge criteria. Statistical process control analysis was utilized to identify system-level changes over time. We used linear regression to assess pre- and postpathway differences in length of stay and transfer to the psychiatry unit. Results: There were 161 patient encounters for anorexia nervosa admitted for medical stabilization. 84% of the sample were female with median age of 15.2 (IQR 14.0-17.0) years. There was no difference in hospital length of stay between the pre- and postpathway groups. There was a statistically significant increase in the proportion of patients transferred to the psychiatry unit over the study period. Conclusion: Clinical pathway use to deliver standardized care to achieve medical stability for patients with anorexia nervosa did not shorten hospital length of stay. Multiple potentially confounding medical and psychosocial factors may have contributed to this lack of improvement.

Differences in Nutrient Intake with Homemade versus Chef-Prepared Specific Carbohydrate Diet Therapy in Inflammatory Bowel Disease: Insights into Dietary Research.

PURPOSE: The aim of this study was to evaluate the nutrient content consumed by children and adolescents on home-prepared versus chef-prepared specific carbohydrate diets (SCD) as therapy for inflammatory bowel disease (IBD). METHODS: Dietary intake of two cohorts with active IBD initiating the SCD over 12 weeks was assessed. The home-prepared cohort received detailed guidance from dietitians on implementation of the SCD. The chef in the other cohort was knowledgeable in the SCD and prepared meals from a fixed set of recipes. Data from 3-day diet diaries at 4 different time points were collected. US Recommended Daily Allowances (RDA) were calculated for macronutrients, vitamins, and minerals. RESULTS: Eight participants on the homemade SCD and 5 participants on the chef-prepared SCD were included in analysis. Mean % RDA for energy intake was 115% and 87% for homemade and chef-prepared groups (p<0.01). Mean % RDA for protein intake was 337% for homemade SCD and 216% for chef-prepared SCD (p<0.01). The homemade SCD group had higher mean % RDA values for vitamin A and iron, while the chef-prepared SCD group had higher intake of vitamins B1, B2, D, phosphorus and zinc (p<0.01 for all). CONCLUSION: The SCD implemented homemade versus chef-prepared can result in significantly different intake of nutrients and this may influence efficacy of this dietary therapy. Meal preparation dynamics and the motivation of families who pursue dietary treatment may play an important role on the foods consumed and the outcomes on dietary therapy with the SCD.

Cryopreserved platelets and their suitability in being re-suspended in additive solution.

BACKGROUND: Cryopreserved platelets are generally produced and stored in plasma. With the advent of additive solutions being routinely used it would be prudent to examine producing and storing these units in a platelet additive solution (SSP+). STUDY DESIGN AND METHODS: Platelet concentrates were prepared from twenty overnight held whole blood units with 10 being re-suspended in 100% plasma and 10 in approximately 70% SSP + and 30% plasma. All had 6% v/v DMSO added prior to storage at -80°C. On thawing plasma stored platelets were re-constituted in fresh plasma with additive prepared platelets being subsequently suspended in 100% SSP+. Sample analysis was assessed pre cryopreservation, post thaw and 4 h. RESULTS: We noted a significant increase in our annexin V levels along with a decrease in GP1bα Von Willebrand binding sites post thaw. The platelets ability to change shape was also significantly reduced as observed with our HSR and ESC values. However, despite this there was still sufficient material within the platelet to allow them to be viable as observed with our thromboelasticity results which, were still observed to be within normal parameters post thaw We also observed an increase in Extracellular vesicles post thaw, suggesting platelet damage which was supported by the reduction in platelet counts. Although there were still sufficient numbers to meet the minimum requirements of the UK guidelines.

Human settlement of East Polynesia earlier, incremental, and coincident with prolonged South Pacific drought.

The timing of human colonization of East Polynesia, a vast area lying between Hawai'i, Rapa Nui, and New Zealand, is much debated and the underlying causes of this great migration have been enigmatic. Our study generates evidence for human dispersal into eastern Polynesia from islands to the west from around AD 900 and contemporaneous paleoclimate data from the likely source region. Lake cores from Atiu, Southern Cook Islands (SCIs) register evidence of pig and/or human occupation on a virgin landscape at this time, followed by changes in lake carbon around AD 1000 and significant anthropogenic disturbance from c. AD 1100. The broader paleoclimate context of these early voyages of exploration are derived from the Atiu lake core and complemented by additional lake cores from Samoa (directly west) and Vanuatu (southwest) and published hydroclimate proxies from the Society Islands (northeast) and Kiribati (north). Algal lipid and leaf wax biomarkers allow for comparisons of changing hydroclimate conditions across the region before, during, and after human arrival in the SCIs. The evidence indicates a prolonged drought in the likely western source region for these colonists, lasting c. 200 to 400 y, contemporaneous with the phasing of human dispersal into the Pacific. We propose that drying climate, coupled with documented social pressures and societal developments, instigated initial eastward exploration, resulting in SCI landfall(s) and return voyaging, with colonization a century or two later. This incremental settlement process likely involved the accumulation of critical maritime knowledge over several generations.

Rapa Nui (Easter Island) monument (ahu) locations explained by freshwater sources.

Explaining the processes underlying the emergence of monument construction is a major theme in contemporary anthropological archaeology, and recent studies have employed spatially-explicit modeling to explain these patterns. Rapa Nui (Easter Island, Chile) is famous for its elaborate ritual architecture, particularly numerous monumental platforms (ahu) and statuary (moai). To date, however, we lack explicit modeling to explain spatial and temporal aspects of monument construction. Here, we use spatially-explicit point-process modeling to explore the potential relations between ahu construction locations and subsistence resources, namely, rock mulch agricultural gardens, marine resources, and freshwater sources-the three most critical resources on Rapa Nui. Through these analyses, we demonstrate the central importance of coastal freshwater seeps for precontact populations. Our results suggest that ahu locations are most parsimoniously explained by distance from freshwater sources, in particular coastal seeps, with important implications for community formation and inter-community competition in precontact times.

Timing of gamma irradiation and blood donor sex influences in vitro characteristics of red blood cells.

BACKGROUND: There are few studies investigating the effect of irradiation on red blood cells (RBCs) during storage. This study analyzed changes in in vitro quality of RBCs irradiated at several points during storage with the aim of providing evidence to support current maximum pre- and postirradiation storage limits. STUDY DESIGN AND METHODS: Each of seven participating centers produced four pools of 7 standard RBC units (SAGM, AS-3, or PAGGSM), which were then split back into 7 units. All units in a pool were from sex-matched blood donors. Every week during 6 weeks of refrigerated storage, 1 unit was irradiated, while 1 unit was not irradiated (control). Units were tested weekly for biochemical variables, morphology, and mechanical fragility. RESULTS: The earlier during storage that units were irradiated, the higher the hemolysis and K+ at end of storage. Irrespective of the timing of irradiation, there was a rapid increase in extracellular K+ , followed by a more gradual increase in hemolysis. ATP levels decreased faster in irradiated units and were reduced below accepted values if irradiated early. Irradiated female RBCs had an absolute lower hemolysis and K+ level compared to male RBCs at all time points. CONCLUSIONS: The method of blood component manufacturing determined the absolute levels of hemolysis and potassium in irradiated and nonirradiated units, but did not influence the effect that timing of irradiation had on the in vitro quality characteristics. This study provides support for the current Council of Europe guidelines on the time limitations for the irradiation of RBCs.

Stool Microbiota at Neutrophil Recovery Is Predictive for Severe Acute Graft vs Host Disease After Hematopoietic Cell Transplantation.

BACKGROUND: Graft-versus-host disease (GVHD) is common after allogeneic hematopoietic cell transplantation (HCT). Risk for death from GVHD has been associated with low bacterial diversity in the stool microbiota early after transplant; however, the specific species associated with GVHD risk remain poorly defined. METHODS: We prospectively collected serial weekly stool samples from 66 patients who underwent HCT, starting pre-transplantation and continuing weekly until 100 days post-transplant, a total of 694 observations in HCT recipients. We used 16S rRNA gene polymerase chain reaction with degenerate primers, followed by high-throughput sequencing to assess the relative abundance of sequence reads from bacterial taxa in stool samples over time. RESULTS: The gut microbiota was highly dynamic in HCT recipients, with loss and appearance of taxa common on short time scales. As in prior studies, GVHD was associated with lower alpha diversity of the stool microbiota. At neutrophil recovery post-HCT, the presence of oral Actinobacteria and oral Firmicutes in stool was positively correlated with subsequent GVHD; Lachnospiraceae were negatively correlated. A gradient of bacterial species (difference of the sum of the relative abundance of positive correlates minus the sum of the relative abundance of negative correlates) was most predictive (receiver operator characteristic area under the curve of 0.83) of subsequent severe acute GVHD. CONCLUSIONS: The stool microbiota around the time of neutrophil recovery post-HCT is predictive of subsequent development of severe acute GVHD in this study.

Evaluating risk factors for Clostridium difficile infection in adult and pediatric hematopoietic cell transplant recipients.

BACKGROUND: Although hematopoietic cell transplant (HCT) recipients are routinely exposed to classic risk factors for Clostridium difficile infection (CDI), few studies have assessed CDI risk in these high-risk patients, and data are especially lacking for pediatric HCT recipients. We aimed to determine incidence and risk factors for CDI in adult and pediatric allogeneic HCT recipients. METHODS: CDI was defined as having diarrhea that tested positive for C. difficile via PCR, cytotoxin assay, or dual enzyme immunoassays. We included all patients who received an allogeneic HCT from 2008 to 2012 at the Fred Hutchinson Cancer Research Center; those <1 year old or with CDI within 8 weeks pre-HCT were excluded. Patients were categorized by transplanting hospital ("adult" or "pediatric") and followed for 100 days post-HCT. RESULTS: Of 1182 HCT recipients, CDI was diagnosed in 17 % (33/192) of pediatric recipients for an incidence of 20 per 10,000 patient-days, and 11 % (107/990) of adult recipients for an incidence of 12 per 10,000. Pediatric recipients were diagnosed a median of 51 days (interquartile range [IQR]: 5, 72) after HCT and adults at 16 days (IQR = 5, 49). Compared with calendar year 2008, pediatric recipients transplanted in 2012 were at increased risk for CDI (hazard ratio [HR] = 3.99, p =.02). Myeloablative conditioning increased CDI risk in adult recipients (HR = 1.81, p =.005). CONCLUSIONS: Pediatric and adult allogeneic recipients are at high risk of CDI post-HCT, particularly adult recipients of myeloablative conditioning. Differences in CDI incidence between children and adults may have resulted from exposure differences related to age; therefore, separately evaluating these groups should be considered in future CDI studies.

Sestrin2 promotes LKB1-mediated AMPK activation in the ischemic heart.

The regulation of AMPK in the ischemic heart remains incompletely understood. Recent evidence implicates the role of Sestrin2 in the AMPK signaling pathway, and it is hypothesized that Sestrin2 plays an influential role during myocardial ischemia to promote AMPK activation. Sestrin2 protein was found to be expressed in adult cardiomyocytes and accumulated in the heart during ischemic conditions. Sestrin2 knockout (KO) mice were used to determine the importance of Sestrin2 during ischemia and reperfusion (I/R) injury. When wild-type (WT) and Sestrin2 KO mice were subjected to in vivo I/R, myocardial infarct size was significantly greater in Sestrin2 KO compared with WT hearts. Similarly, Langendorff perfused hearts indicated exacerbated postischemic contractile function in Sestrin2 KO hearts compared with WT. Ischemic AMPK activation was found to be impaired in the Sestrin2 KO hearts. Immunoprecipitation of Sestrin2 demonstrated an association with AMPK. Moreover, liver kinase B1 (LKB1), a major AMPK upstream kinase, was associated with the Sestrin2-AMPK complex in a time-dependent manner during ischemia, whereas this interaction was nearly abolished in Sestrin2 KO hearts. Thus, Sestrin2 plays an important role in cardioprotection against I/R injury, serving as an LKB1-AMPK scaffold to initiate AMPK activation during ischemic insults.

Impaired SIRT1 nucleocytoplasmic shuttling in the senescent heart during ischemic stress.

A "longevity " gene, sirtuin 1 (SIRT1), can attenuate age-dependent induction of left ventricular dysfunction. This study aimed to characterize the role of SIRT1 in the tolerance of aged heart to ischemic insults. Male C57BL/6 young (4-6 mo) and aged (24-26 mo) mice were used to determine the role of SIRT1 in myocardial ischemia/reperfusion (I/R) tolerance. SIRT1 localization was assessed by confocal microscopy. Immunoblotting was used to evaluate SIRT1 expression and translocation. The results demonstrated that SIRT1 is expressed predominantly as a sumoylated form in cardiomyocyte nuclei. Moreover, cardiac overexpression of desumoylase, sentrin-specific protease 2 (SENP2), significantly reduces nuclear sumoylated SIRT1 levels (P<0.05). Interestingly, I/R stress leads to desumoylation and translocation of nuclear SIRT1 into the cytoplasm in aged but not in young hearts. SIRT1 activity in ischemic young hearts was 3.2-fold higher than that seen in ischemic aged hearts, which suggests that aging causes impaired nucleocytoplasmic shuttling and activation of SIRT1 during ischemic stress. The infarct size in aged and Sirt1(+/-) knockout hearts was higher than that observed in young and Sirt1(+/+) WT littermate hearts, respectively (all P<0.05). SIRT1 agonist, SRT1720, reduced myocardial infarction in both aged and Sirt1(+/-) hearts. Therefore, impaired cardiac SIRT1 activity plays a critical role in the observed increase in susceptibility of the aged heart to I/R injury. SIRT1 agonist can restore this aging-related loss of cardioprotection.

Alternol exerts prostate-selective antitumor effects through modulations of the AMPK signaling pathway.

BACKGROUND: Alternol is an original compound purified from the fermentation products of Alternaria alternata var. monosporus, a microorganism from the bark of the yew tree. It has been reported that Alternol can inhibit proliferation of mouse leukemia cells and human gastric carcinoma cells, the aim of this study was to investigate the effects of Alternol on prostate cancer cells in comparison to prostate cells. METHODS: The MTT assay was utilized to assess cell viability. Cell cycle was analyzed by flow cytometry with propidium iodide staining. Protein expression levels were examined by Western blotting. RESULTS: Alternol treatment resulted in a significant decrease in the viability of prostate cancer cells but had lesser effects on prostate cells. Alternol inhibited AMP-activated protein kinase (AMPK) phosphorylation in prostate cancer C4-2 cells but stimulated AMPK phosphorylation in prostate RWPE-1 cells. Inhibition of p27 phosphorylation was observed in C4-2 cells whereas a promotion of p27 phosphorylation was seen in RWPE-1 cells. Alternol treatment resulted in a profound increase in the LC3II/LC3I protein ratio in RWPE-1 cells but not in C4-2 cells. A dose-dependent down-regulation of Bcl-2 protein was detected in C4-2 cells but not in RWPE-1 cells. Pretreatment of cells with Compound C (AMPK inhibitor) before Alternol treatment abolished the selective antitumor effect of Alternol. CONCLUSIONS: These results reveal for the first time that Alternol exerts a selective antitumor effect on prostate cancer cells when compared with RWPE-1 prostate epithelial cells. In addition, the AMPK signaling pathway is responsible for the selective antitumor effects of Alternol.

Polymers for the rapid and effective activation and aggregation of platelets.

Platelets are responsible for plugging sites of vascular injury, where upon activation they spread out and become cross-linked, preventing further blood loss. It is desirable to control the activation process on demand for applications such as the rapid staunching of blood flow following trauma. Polymers are the material of choice in many biological areas, with physical properties that allow control of morphology as well as ease of functionalisation and production. Herein, polymer microarrays were used to screen a complex human fluid (platelet rich plasma) to identify polyacrylates that could be used to modulate platelet activation. Several polymers were identified which rapidly activated platelets as determined by CD61P binding and subsequent confirmation by scanning electron microcopy analysis. This approach enabled a direct comparison between the natural agonist collagen and synthetic polymers with respect to the activation status of the platelets as well as the number of bound platelets. Further investigations under physiological flow demonstrated that the static microarray experiments gave viable candidates for potential medical applications while specific protein binding to the polymers was identified as a possible mode of action. The approach demonstrates the ability of polymer microarrays to identify new polymers for specific biological activation events and in this case allowed the identification of materials that allowed higher levels of platelets to bind in advanced activation states than the natural standard collagen in static and flow studies.

Acute rosiglitazone treatment is cardioprotective against ischemia-reperfusion injury by modulating AMPK, Akt, and JNK signaling in nondiabetic mice.

Rosiglitazone (RGZ), a peroxisome proliferator-activated receptor (PPAR)-γ agonist, has been demonstrated to possess cardioprotective properties during ischemia-reperfusion. However, this notion remains controversial as recent evidence has suggested an increased risk in cardiac events associated with long-term use of RGZ in patients with type 2 diabetes. In this study, we tested the hypothesis that acute RGZ treatment is beneficial during I/R by modulating cardioprotective signaling pathways in a nondiabetic mouse model. RGZ (1 µg/g) was injected intravenously via the tail vein 5 min before reperfusion. Myocardial infarction was significantly reduced in mice treated with RGZ compared with vehicle controls (8.7% ± 1.1% vs. 20.2% ± 2.5%, P < 0.05). Moreover, isolated hearts were subjected to 20 min of global, no-flow ischemia in an ex vivo heart perfusion system. Postischemic recovery was significantly improved with RGZ treatment administered at the onset of reperfusion compared with vehicle (P < 0.001). Immunoblot analysis data revealed that the levels of both phospho-AMP-activated protein kinase (Thr(172)) and phospho-Akt (Ser(473)) were significantly upregulated when RGZ was administered 5 min before reperfusion compared with vehicle. On the other hand, inflammatory signaling [phospho-JNK (Thr(183)/Tyr(185))] was significantly downregulated as a result of RGZ treatment compared with vehicle (P < 0.05). Intriguingly, pretreatment with the selective PPAR-γ inhibitor GW-9662 (1 µg/g iv) 10 min before reperfusion significantly attenuated these beneficial effects of RGZ on the ischemic heart. Taken together, acute treatment with RGZ can reduce ischemic injury in a nondiabetic mouse heart via modulation of AMP-activated protein kinase, Akt, and JNK signaling pathways, which is dependent on PPAR-γ activation.

PPAR-γ and AMPK--advantageous targets for myocardial ischemia/reperfusion therapy.

Ischemic heart disease stands as the number one leading cause of death in the United States. Current interventions rely on the immediate restoration of blood flow to the ischemic area; however, this in turn may trigger a series of undesirable events that are further injurious to the myocardium, termed ischemia/reperfusion (I/R) injury. Therefore, there is a need for novel therapeutic strategies aimed at limiting the extent of myocardial injury. Yet, the molecular mechanisms responsible for I/R injury remain largely indefinable. Research efforts are currently investigating various signaling mechanisms to be used for potential targets limiting cardiac injury due to such cardiovascular events. In this review, we highlight two potential molecular targets, PPAR-γ and AMPK, which have been extensively reported to have various cardioprotective capabilities against I/R injury. Although functionally different, the pathways these proteins mediate seem to intersect and possibly act synergistically potentiating a cardioprotective response.

Characterization of a monoclonal antibody to erythroid related factor.

We describe here a novel IgG monoclonal antibody to erythroid-related factor (ERAF), also known as alpha hemoglobin stabilizing protein (AHSP) and eryththroid differentiation related factor (EDRF). Our antibody named PCE 5 is an IgG(1) kappa chain and is to the peptide sequence MVTVVE ranked highly in our active site analysis and binds with high affinity to ERAF.

Fluoride induces apoptosis and alters collagen I expression in rat osteoblasts.

In this study we investigated apoptosis and expression of the collagen I gene in newborn rat osteoblasts (OB) by the administration of varying concentrations of fluoride (F). Sodium fluoride (NaF) at concentrations of 0, 0.5, 5, 10, and 20mg/L was administered to cultured OB. The percentage of G(1)/G(0) (Gap 1/Gap 0), S (synthesis), G(2)/M (Gap 2/M, mitosis), and apoptosis rates in OB were analyzed with a Fluorescence-activated Cell Sorter (FACS) by propidium iodine (PI) staining after F treatment of 72 h. Effects of F treatment on COL1A1 and COL1A2 mRNA and collagen I protein levels were determined using quantitative real time RT-PCR (qRT-PCR) and immunofluorescence, respectively. This study demonstrates that there is a pronounced negative effect of long term NaF treatment on OB survival. These negative effects include an inhibition of the transformation from S phase into G(2)/M phase, increased apoptosis, and decreased COL1A1 mRNA, down-regulating the synthesis of COL I protein. The results suggest that COL I protein degradation in OB from F toxicity is due to a depletion of COL1A1 mRNA and not COL1A2.

Macrophage migration inhibitory factor deficiency augments cardiac dysfunction in Type 1 diabetic murine cardiomyocytes.

BACKGROUND: It has become evident that macrophage migration inhibitory factor (MIF) is associated with the development of Type 1 diabetes mellitus. The aim of the present study was to determine whether MIF plays a role in cardiac contractile dysfunction in T1DM mice. METHODS: Mechanical and intracellular Ca(2+) properties were measured in cardiomyocytes isolated from wild-type (WT) and MIF-knockout (MIF-KO) mice administrated or not streptozotocin (200 mg/kg, i.p.). Relative stress signaling was evaluated using western blot analysis. RESULTS: Peak shortening (PS) and maximal velocity of shortening/relengthening (±dL/dt) were reduced and the duration of relengthening (TR90) was prolonged in both WT and MIF-KO cardiomyocytes treated with STZ (P < 0.01 vs control), which may be associated with reduced intracellular Ca(2+) decay in both groups. However, STZ-treated WT cardiomyocytes demonstrated significantly better contractile function and intracellular Ca(2+) properties compared with STZ-treated MIF-KO cardiomyocytes (all P < 0.05). Interestingly, the physiological data clearly showed that blood glucose levels were significantly higher in STZ-treated MIF-KO mice than STZ-treated WT mice (P < 0.01). Moreover, phosphorylation of AMP-activated protein kinase (AMPK) and its direct downstream target acetyl-CoA carboxylase (ACC) was markedly lower in hearts from STZ-treated MIF-KO mice than STZ-treated WT mice (P < 0.05). There were no significant differences between untreated WT and MIF-KO control groups. CONCLUSIONS: There is a beneficial action of MIF in the management of cardiac dysfunction in T1DM. The cardioprotective effect of MIF may be associated with AMPK signaling.

Pathogen reduction of fresh plasma using riboflavin and ultraviolet light: effects on plasma coagulation proteins.

BACKGROUND: Treatment with riboflavin and ultraviolet (UV) light reduces the pathogens present in blood components. This study assessed changes to the coagulation proteins that had occurred during this treatment of fresh plasma units before freezing. STUDY DESIGN AND METHODS: Twenty fresh plasma units (230 +/- 30 mL) were treated by the Mirasol process (CaridianBCT Biotechnologies) and frozen within 8 hours of donation. Plasma units were combined with 35 mL of a 500 micromol/L riboflavin solution in an illumination bag to achieve a final concentration of approximately 60 micromol/L riboflavin. The bag was placed in the Mirasol illuminator and exposed to UV light (6.24 J/mL). Samples were frozen before and after treatment. RESULTS: Recoveries observed were 67.7 +/- 3.9% Factor (F)XI, 68.5 +/- 3.3% FVIII:C, 78.8 +/- 4.5% fibrinogen, 78.9 +/- 4.1% FV, 79.0 +/- 4.2% FVII, 79.0 +/- 8.6% F IX, 79.7 +/- 2.6% FX, and 85.0 +/- 3.7% FII. Von Willebrand factor (VWF) antigen, VWF:ristocetin cofactor, and ADAMTS13 recoveries were 87.0 +/- 7.1, 85.5 +/- 6.6, and 73.3 +/- 15.2%, respectively, while that of protein C was 83.6 +/- 2.6%. A loss of high-molecular-weight VWF multimers was observed in most units. Recoveries for protein S, antithrombin, and plasmin inhibitor were greater than 90%. The mean FVIII:C concentration, after treatment, was 0.76 +/- 0.17 IU/mL. CONCLUSIONS: As with other pathogen reduction technologies, the Mirasol process resulted in some loss of coagulation factor activity. For most Mirasol-treated units and for most of the tested factors this is unlikely to have clinical impact, but trials are required to demonstrate this.

Freezing of buffy coat-derived, leukoreduced platelet concentrates in 6 percent dimethyl sulfoxide.

BACKGROUND: There has recently been renewed interest in freezing platelets (PLTs) in dimethyl sulfoxide (DMSO) for the treatment of major traumatic injuries, especially in military situations. This study examined PLTs that were frozen in small volumes of 6 percent DMSO at -80 degrees C. STUDY DESIGN AND METHODS: Buffy coat-derived pooled leukoreduced PLT concentrates were frozen in 6 percent DMSO and stored at -80 degrees C. Assays included hypotonic shock response (HSR); aggregation; glycoprotein (GP)Ibalpha and P-selectin binding sites; annexin V binding to phosphatidylserine, glycocalicin, and lactate dehydrogenase (LDH). Cone and plate technology (DiaMed Impact-R, DiaMed) was used to test PLT function under near physiologic conditions. RESULTS: The freeze-thaw loss of PLTs was 23 percent. HSR was 17 +/- 7 percent. Cytometry demonstrated two populations of PLTs: one with normal levels of GPIbalpha binding sites (27 x 10(3) +/- 3 x 10(3)/PLT) and one with reduced levels (5.5 x 10(3) +/- 1.2 x 10(3)/PLT). There were 1.4 x 10(3) +/- 0.2 x 10(3) P-selectin binding sites per PLT. Annexin V binding to phosphatidylserine was 50 +/- 9 percent and LDH was 496 +/- 207 IU per 10(12) PLTs. Surface coverage and aggregate size, as measured by the DiaMed Impact-R, were similar to those observed with PLTs stored for 2 days at 22 degrees C. CONCLUSION: Some degree of activation was demonstrated by the proportion of PLTs with reduced levels of GPIbalpha binding sites, increased P-selectin expression, and increased Annexin V binding. LDH concentrations indicated a degree of lysis. The DiaMed Impact-R results showed that the PLTs were still capable of adhering to surfaces and forming aggregates under shear force.