Effects of pathogen reduction technology and storage duration on the ability of cryoprecipitate to rescue induced coagulopathies in vitro

BACKGROUND: Fibrinogen concentrates and cryoprecipitate are currently used for fibrinogen supplementation in bleeding patients with dysfibrinogenemia. Both products provide an abundant source of fibrinogen but take greater than 10 min to prepare for administration. Fibrinogen concentrates lack coagulation factors (i.e., factor VIII [FVIII], factor XIII [FXIII], von Willebrand factor [VWF]) important for robust hemostatic function. Cryoprecipitate products contain these factors but have short shelf lives (\u3c6 h). Pathogen reduction (PR) of cryoprecipitate would provide a shelf-stable immediately available adjunct containing factors important for rescuing hemostatic dysfunction. STUDY DESIGN AND METHODS: Hemostatic adjunct study products were psoralen-treated PR-cryoprecipitated fibrinogen complex (PR-Cryo FC), cryoprecipitate (Cryo), and fibrinogen concentrates (FibCon). PR-Cryo FC and Cryo were stored for 10 days at 20-24°C. Adjuncts were added to coagulopathies (dilutional, 3:7 whole blood [WB]:normal saline; or lytic, WB + 75 ng/ml tissue plasminogen activator), and hemostatic function was assessed by rotational thromboelastometry and thrombin generation. RESULTS: PR of cryoprecipitate did not reduce levels of FVIII, FXIII, or VWF. PR-Cryo FC rescued dilutional coagulopathy similarly to Cryo, while generating significantly more thrombin than FibCon, which also rescued dilutional coagulopathy. Storage out to 10 days at 20-24°C did not diminish the hemostatic function of PR-Cryo FC. DISCUSSION: PR-Cryo FC provides similar and/or improved hemostatic rescue compared to FibCon in dilutional coagulopathies, and this rescue ability is stable over 10 days of storage. In hemorrhaging patients, where every minute delay is associated with a 5% increase in mortality, the immediate availability of PR-Cryo FC has the potential to improve outcomes

Using wildlife activity and antibiotic resistance analysis to model bacterial water quality in coastal ponds

Models that help predict fecal coliform bacteria (FCB) levels in environmental waters can be important tools for resource managers. In this study, we used animal activity along with antibiotic resistance analysis (ARA), land cover, and other variables to build models that predict bacteria levels in coastal ponds that discharge into an estuary. Photographic wildlife monitoring was used to estimate terrestrial and aquatic wildlife activity prior to sampling. Increased duck activity was an important predictor of increased FCB in coastal ponds. Terrestrial animals like deer and raccoon, although abundant, were not significant in our model. Various land cover types, rainfall, tide, solar irradiation, air temperature, and season parameters, in combination with duck activity, were significant predictors of increased FCB. It appears that tidal ponds allow for settling of bacteria under most conditions. We propose that these models can be used to test different development styles and wildlife management techniques to reduce bacterial loading into downstream shellfish harvesting and contact recreation areas

Genetic Tests To Evaluate Prognosis and Predict Therapeutic Response in Acute Myeloid Leukemia

Management of patients with acute myeloid leukemia relies on genetic tests that inform diagnosis and prognosis, predict response to therapy, and measure minimal residual disease. The value of genetics is reinforced in the revised 2008 World Health Organization acute myeloid leukemia classification scheme. The various analytic procedures—karyotype, fluorescence in situ hybridization, reverse transcription polymerase chain reaction, DNA sequencing, and microarray technology—each have advantages in certain clinical settings, and understanding their relative merits assists in specimen allocation and in effective utilization of health care resources. Karyotype and array technology represent genome-wide screens, whereas the other methods target specific prognostic features such as t(15;17) PML-RARA, t(8;21) RUNX1-RUNX1T1, inv(16) CBFB-MYH11, 11q23 MLL rearrangement, FLT3 internal tandem duplication, or NPM1 mutation. New biomarkers and pharmacogenetic tests are emerging. The pathologist's expertise is critical in 1) consulting with clinicians about test selection as well as specimen collection and handling; 2) allocating tissue for immediate testing and preserving the remaining specimen for any downstream testing that is indicated once morphology and other pertinent test results are known; 3) performing tests that maximize outcome based on the strengths and limitations of each assay in each available specimen type; and 4) interpreting and conveying results to the rest of the health care team in a format that facilitates clinical management. Acute myeloid leukemia leads the way for modern molecular medicine

Chemotherapy and Stem Cell Transplantation Increase p16

AbstractThe expression of markers of cellular senescence increases exponentially in multiple tissues with aging. Age-related physiological changes may contribute to adverse outcomes in cancer survivors. To investigate the impact of high dose chemotherapy and stem cell transplantation on senescence markers in vivo, we collected blood and clinical data from a cohort of 63 patients undergoing hematopoietic cell transplantation. The expression of p16INK4a, a well-established senescence marker, was determined in T-cells before and 6months after transplant. RNA sequencing was performed on paired samples from 8 patients pre- and post-cancer therapy. In patients undergoing allogeneic transplant, higher pre-transplant p16INK4a expression was associated with a greater number of prior cycles of chemotherapy received (p=0.003), prior autologous transplantation (p=0.01) and prior exposure to alkylating agents (p=0.01). Transplantation was associated with a marked increase in p16INK4a expression 6months following transplantation. Patients receiving autologous transplant experienced a larger increase in p16INK4a expression (3.1-fold increase, p=0.002) than allogeneic transplant recipients (1.9-fold increase, p=0.0004). RNA sequencing of T-cells pre- and post- autologous transplant or cytotoxic chemotherapy demonstrated increased expression of transcripts associated with cellular senescence and physiological aging. Cytotoxic chemotherapy, especially alkylating agents, and stem cell transplantation strongly accelerate expression of a biomarker of molecular aging in T-cells

Application of a Near Infrared Imaging System for Thermographic Imaging of the Space Shuttle during Hypersonic Re-Entry

High resolution calibrated near infrared (NIR) imagery was obtained of the Space Shuttle s reentry during STS-119, STS-125, and STS-128 missions. The infrared imagery was collected using a US Navy NP-3D Orion aircraft using a long-range infrared optical package referred to as Cast Glance. The slant ranges between the Space Shuttle and Cast Glance were approximately 26-41 nautical miles at point of closest approach. The Hypersonic Thermodynamic Infrared Measurements (HYTHIRM) project was a NASA Langley led endeavor sponsored by the NASA Engineering Safety Center, the Space Shuttle Program Office and the NASA Aeronautics Research Mission Directorate to demonstrate a quantitative thermal imaging capability. HYTHIRM required several mission tools to acquire the imagery. These tools include pre-mission acquisition simulations of the Shuttle trajectory in relationship to the Cast Glance aircraft flight path, radiance modeling to predict the infrared response of the Shuttle, and post mission analysis tools to process the infrared imagery to quantitative temperature maps. The spatially resolved global thermal measurements made during the Shuttle s hypersonic reentry provides valuable flight data for reducing the uncertainty associated with present day ground-to-flight extrapolation techniques and current state-of-the-art empirical boundary-layer transition or turbulent heating prediction methods. Laminar and turbulent flight data is considered critical for the development of turbulence models supporting NASA s next-generation spacecraft. This paper will provide the motivation and details behind the use of an upgraded NIR imaging system used onboard a Navy Cast Glance aircraft and describe the characterizations and procedures performed to obtain quantitative temperature maps. A brief description and assessment will be provided of the previously used analog NIR camera along with image examples from Shuttle missions STS-121, STS-115, and solar tower test. These thermal observations confirmed the challenge of a long-range acquisition during re-entry. These challenges are due to unknown atmospheric conditions, image saturation, vibration etc. This provides the motivation for the use of a digital NIR sensor. The characterizations performed on the digital NIR sensor included radiometric, spatial, and spectral measurements using blackbody radiation sources and known targets. An assessment of the collected data for three Space Shuttle atmospheric re-entries, STS-119, STS-125, and STS-128, are provided along with a description of various events of interest captured using the digital NIR imaging system such as RCS firings and boundary layer transitions. Lastly the process used to convert the raw image counts to quantitative temperatures is presented along with comparisons to the Space Shuttle's onboard thermocouples

Multifactorial resistance to adriamycin: relationship of DNA repair, glutathione transferase activity, drug efflux, and p-glycoprotein in cloned cell lines of adriamycin-sensitive and -resistant P388 Leukemia

Cloned lines of Adriamycin (ADR)-sensitive and -resistant P388 leukemia have been established, including P388/ADR/3 and P388/ADR/7 that are 5- and 10-fold more resistant than the cloned sensitive cell line P388/4 (Cancer Res., 46: 2978, 1986). A time course of ADR-induced DNA double-strand breaks revealed that in sensitive P388/4 cells, evidence of DNA repair was noted 4 h after removal of drug, whereas in resistant clone 3 and 7 cells repair was observed 1 h after drug removal. The earlier onset of DNA repair was statistically significant (p = 0.0154 for clone 3 cells, and p = 0.0009 for clone 7 cells). By contrast, once the repair process was initiated, the rate of repair was similar for all three cell lines. The level of glutathione transferase activity was determined in whole cell extracts. Enzyme activity (mean ± SE) in sensitive cells was 9.49 ± 1.00 nmol/min/mg protein, that in resistant clone 3 cells was 13.36 ± 1.03 nmol/min/mg, and that in clone 7 cells was 13.96 ± 1.44 nmol/min/mg; the 1.44-fold increase in enzyme activity in resistant cells was statistically significant (p = 0.01). Further evidence of induction of glutathione transferase was provided by Northern blot analysis using a 32P-labeled cDNA for an anionic glutathione transferase, which demonstrated approximately a twofold increase in mRNA in resistant clone 7 cells. Western blot analysis with a polyvalent antibody against anionic glutathione transferase also revealed a proportionate increase in gene product in resistant cells. Dose-survival studies showed that ADR-resistant cells were cross-resistant to actinomycin D, daunorubicin, mitoxantrone, colchicine, and etoposide, but not to the alkylating agent melphalan; this finding provided evidence that these cells are multidrug resistant. Using a cDNA probe for P-glycoprotein, a phenotypic marker for multidrug resistance, Northern blot analysis showed an increase in the steady state level of mRNA of approximately twofold in resistant clone 3 and 7 cells. Southern analysis with the same cDNA probe showed no evidence of gene amplification or rearrangement. Western blot analysis with monoclonal C219 antibody demonstrated a distinct increase in P-glycoprotein in resistant cells. Efflux of Adriamycin as measured by the efflux rate constant was identical in all three cell lines. Furthermore, the metabolic inhibitors azide and dinitrophenol did not augment drug uptake in either sensitive or resistant cells. These findings suggest that despite the increase in P-glycoprotein, an active extrusion pump was not operational in these cells. This and previous studies provide unequivocal evidence that resistance to Adriamycin is multifactorial. Decreased drug uptake, decreased formation of DNA single- and double-strand breaks, increased glutathione transferase activity, earlier onset of DNA repair, as well as elevated P-glycoprotein are all characteristic of multifactorial drug resistance

Dose-dependent von Willebrand Factor inhibition by aptamer BB-031 correlates with thrombolysis in a microfluidic model of arterial occlusion

Von Willebrand Factor (VWF) plays a critical role in thrombus formation, stabilization, and propagation. Previous studies have demonstrated that targeted inhibition of VWF induces thrombolysis when administered in vivo in animal models of ischemic stroke. The study objective was to quantify dose-dependent inhibition of VWF-platelet function and its relationship with thrombolysis using BB-031, an aptamer that binds VWF and inhibits its function. VWF:Ac, VWF:RCo, T-TAS, and ristocetin-induced impedance aggregometry were used to assess BB-031-mediated inhibition of VWF. Reductions in original thrombus surface area and new deposition during administration of treatment were measured in a microfluidic model of arterial thrombolysis. Rotational thromboelastometry was used to assess changes in hemostasis. BB-031 induced maximal inhibition at the highest dose (3384 nM) in VWF:Ac, and demonstrated dose-dependent responses in all other assays. BB-031, but not vehicle, induced recanalization in the microfluidic model. Maximal lytic efficacy in the microfluidic model was seen at 1692 nM and not 3384 nM BB-031 when assessed by surface area. Minor changes in ROTEM parameters were seen at 3384 nM BB-031. Targeted VWF inhibition by BB-031 results in clinically measurable impairment of VWF function, and specifically VWF-GPIb function as measured by VWF:Ac. BB-031 also induced thrombolysis as measured in a microfluidic model of occlusion and reperfusion. Moderate correlation between inhibition and lysis was observed. Additional studies are required to further examine off-target effects of BB-031 at high doses, however, these are expected to be above the range of clinical targeted dosing

Microfluidics for the detection of minimal residual disease in acute myeloid leukemia patients using circulating leukemic cells selected from blood

Microfluidic assay for the selection of circulating leukemic cells from peripheral blood for the early detection of minimal residual disease in acute myeloid leukemia patients

The Effects of an Exercise Program in Leukemia Patients

PURPOSE: To examine the feasibility of administering an in-hospital exercise program to acute leukemia patients undergoing chemotherapy. A secondary purpose explored the impact of exercise on selected physiological, psychological, and inflammatory markers. METHODS: Ten patients, aged 18 to 50 years, diagnosed with acute leukemia or newly relapsed were assessed for body weight, height, body composition (skinfolds), cardiorespiratory endurance (total minutes on bicycle ergometer at 60% heart rate reserve), dynamic muscular endurance (Rocky Mountain Cancer Rehabilitation Institute protocol), fatigue (Revised Piper Fatigue Scale), depression (Center for Epidemiologic Studies Depression scale, National Institute of Mental Health questionnaire), and quality of life (Functional Assessment of Cancer Therapy-General) at baseline (within 3 days of diagnosis) and at the end of induction phase of treatment. Blood draws were taken at baseline, midpoint, and at the end of induction for analyses of inflammatory markers (Linco Luminex assay). Combined aerobic and strength training exercises were administered 3 times per week, twice daily, for 30 minutes. Paired-samples t-tests were used for the analyses of physiological and psychological parameters. One-way repeated measures analysis of variance was used for the analyses of inflammatory markers. RESULTS: Significant improvements in cardiorespiratory endurance (P = .009, baseline 8.9 +/- 8.8 minutes, postexercise intervention 17 +/- 14.3 minutes) with significant reductions in total fatigue scores (P = .009, baseline 4.6 +/- 1.7, postexercise intervention 1.8 +/- 1.6) and depression scores (P = .023, baseline 19 +/- 11.5, postexercise intervention 12 +/- 8.2) were observed. Marginally significant decrease in interleukin-6 (IL-6; P = .059) with no significant changes in IL-10 (P = .223) or interferon-gamma (P = .882) were observed. CONCLUSION: Administration of exercise to acute leukemia patients undergoing treatment is feasible. The exercise protocol used increased cardiovascular endurance, reduced fatigue and depression scores, and maintained quality of life. Although no significant change in inflammation was observed, a trend demonstrating a reduction in IL-6 and an increase in IL-10 warrants further investigation

Vitamin D in the general population of young adults with autism in the Faroe Islands

Vitamin D deficiency has been proposed as a possible risk factor for developing autism spectrum disorder (ASD). 25-Hydroxyvitamin D3 (25(OH)D3) levels were examined in a cross-sectional population-based study in the Faroe Islands. The case group consisting of a total population cohort of 40 individuals with ASD (aged 15–24 years) had significantly lower 25(OH)D3 than their 62 typically-developing siblings and their 77 parents, and also significantly lower than 40 healthy age and gender matched comparisons. There was a trend for males having lower 25(OH)D3 than females. Effects of age, month/season of birth, IQ, various subcategories of ASD and Autism Diagnostic Observation Schedule score were also investigated, however, no association was found. The very low 25(OH)D3 in the ASD group suggests some underlying pathogenic mechanism