Kinetics and stoichiometry of reactions between the 2,2-diphenyl-1-picrylhydrazyl (DPPH) stable radical and 25 antioxidant compounds with different structure, molecular weight, number of −OH groups, ...and redox potential were investigated by recording the loss of DPPH• absorbance at 515 nm continuously for 10 min. A series of antioxidant concentrations was tested to determine linear response ranges and reaction saturation points. The primary feature distinguishing antioxidant activityrate of initial reaction (<30 s)was controlled by whether the dominant antioxidant mechanism was electron (very fast) or hydrogen atom (slow) transfer and by impairment of steric accessibility to the DPPH radical site by bulky ring adducts and multiple phenolic rings. Results raise serious questions regarding application of the DPPH assay for ranking antioxidants and natural extracts and suggest possible redirection of this assay to distinguish active reaction mechanisms by comparing reactions rates and patterns in different solvents and in 50% water/methanol mixtures at different pH values.
Emerging evidence is elucidating how non-nutrient phytochemicals underlie the health promotion afforded by fruits and vegetables. This review focuses on Vaccinium macrocarpon, the American cranberry, ...compiling a comprehensive list of its known phytochemical components, and detailing their prevalence in cranberry fruit and its products. Flavonoids, especially colored anthocyanins, abundant flavonols, and unique proanthocyanidins, have attracted major research attention. Other notable active components include phenolic acids, benzoates, hydroxycinnamic acids, terpenes and organic acids. Health effects of cranberries, cranberry products, and isolated cranberry components in humans and animals, as well as in vitro, are debated. Evidence for protection from several bacterial pathogens, cancer, cardiovascular disease, and inflammation is compelling, while neuroprotection and anti-viral activity also have begun to draw new consideration. Emerging bioavailability data is considered and potential molecular mechanisms are evaluated, linking phytochemicals to health effects through their biochemical properties and reactions. Finally, the effects of processing and storage on cranberry phytochemicals is discussed, with a focus on identifying research gaps and novel means to preserve their natural, health-promoting components.
Reaction kinetics in the Trolox equivalent antioxidant capacity (TEAC) assay between ABTS+• 2,2′-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) cation radical and compounds with different ...structure, molecular weight, number of OH groups, and redox potential were investigated by recording loss of ABTS+• absorbance (734 nm) continuously over time. Curves showed six distinguishable kinetic patterns, including both immediate and extended reaction components. Radical quenching rates in the immediate component most relevant to reactions in foods and tissues depended on phenol structure and steric accessibility to the hindered radical, while reaction stoichiometry correlated with the number of phenol groups (>0.81) but not redox potential. Current assay procedures measure antioxidant capacity under conditions not relevant to actual applications and do not determine radical quenching rates. Results raise serious questions regarding the ability of reactions with the hindered ABTS+• to rank actual radical quenching by compounds with different structures and invalidate reporting antioxidant activity as Trolox equivalents.
The spread of bacterial resistance against conventional antibiotics generates a great need for the discovery of novel antimicrobials. Polypeptide antibiotics constitute a promising class of ...antimicrobial agents that favour attack on bacterial membranes. However, efficient measurement platforms for evaluating their mechanisms of action in a systematic manner are lacking. Here we report an integrated lab-on-a-chip multilayer microfluidic platform to quantify the membranolytic efficacy of such antibiotics. The platform is a biomimetic vesicle-based screening assay, which generates giant unilamellar vesicles (GUVs) in physiologically relevant buffers on demand. Hundreds of these GUVs are individually immobilised downstream in physical traps connected to separate perfusion inlets that facilitate controlled antibiotic delivery. Antibiotic efficacy is expressed as a function of the time needed for an encapsulated dye to leak out of the GUVs as a result of antibiotic treatment. This proof-of-principle study probes the dose response of an archetypal polypeptide antibiotic cecropin B on GUVs mimicking bacterial membranes. The results of the study provide a foundation for engineering quantitative, high-throughput microfluidics devices for screening antibiotics.
Immense gains in understanding of mechanisms and effects of lipid oxidation have been achieved in the nearly 90 years over which lipid oxidation has been an active research focus. Even so, the ...substantial questions still being raised about lipid oxidation in this special issue show clearly that missing pieces remain and must be considered for full accounting of this important reaction in any system. In this context, epoxides are spotlighted as a critical overlooked product of lipid autoxidation - underestimated in analysis, underestimated in presence as a functionally active and competitive intermediate and product of lipid oxidation, and underestimated in potential contributions to impact of lipid oxidation on other molecules and cell functions. Logical reasons for ignoring or not finding epoxides are offered in historical development of lipid oxidation knowledge. Reactions generating lipid epoxides in autoxidation are reviewed, limitations in detecting and tracking epoxides are outlined to explain why epoxides may not be detected when they should be present, and justifications for increased research and analysis of epoxides are argued. The main goal is to provide a context for recognizing epoxides as critical products that must be accounted for in determining the state rather than extent of lipid oxidation and in tracking its consequences in oils, foods, personal care products, and tissues. A secondary goal is to stimulate new research using contemporary analyses to fill in the gaps of knowledge about epoxide formation, structure, and reactions in lipid autoxidation.
According to WHO 2008 guidelines, the required percentage of cells manifesting dysplasia in the bone marrow to qualify as significant is 10% or over in one or more hematopoietic cell lineages, but ...this threshold is controversial. No 'normal' values have been established. Therefore, we investigated dyshematopoiesis in bone marrow aspirate squash preparations of 120 healthy bone marrow donors.
Bone marrow squash slides of 120 healthy unrelated bone marrow donors were examined independently by 4 experienced morphologists. Samples were taken from the first aspiration during the harvest. Bone marrow preparation and assessment were performed according to WHO recommendations and ICSH guidelines.
More than 10% dysmyelopoiesis could be detected in 46% of bone marrow aspirate squash preparations with 26% in 2 or more cell lineages and 7% in 3 cell lineages in healthy bone marrow donors. Donors under the age of 30 years exhibited more dysgranulopoietic changes and dysmegakaryopoietic changes (P<0.001) compared to the older donors. Female donors showed more dysgranulopoietic changes than male donors (P = 0.025). The concordance rate between the 4 investigators was modest in dysgranulopoiesis but poor in dyserythropoiesis and dysmegakaryopoiesis.
The poor reliability of the 10% cut off was partly related to the proximity of the current criteria to the observed cut-off mean values of the normal population. These findings question the current WHO threshold of the 10% or over necessary for the percentage of cells manifesting dysplasia to be considered significant, and suggest that either a higher threshold would be more appropriate or different thresholds should be set for each lineage.
Accelerated solvent extraction (ASE) was evaluated for extracting lipids from baked and extruded dry pet foods to determine factors controlling extraction efficiency and effects on lipid oxidation. ...Hydroperoxide decomposition and new lipid oxidation were minimal at 40 °C but increased at higher extraction temperatures without increasing yields. Maximum extraction required grinding samples to 250 µm particles, presence of polar solvents chloroform, chloroform/methanol 2:1 (v/v) mixed, hexane/methanol 2:1 (v/v), and a minimum of 20 min total static extraction time in repeat extraction cycles. Hexane and methanol injected into extraction cells simultaneously but separately was able to nearly duplicate extractions of chloroform/methanol, providing an option for replacing toxic chlorinated hydrocarbon solvents in ASE. However, lipid oxidation was higher in hexane. Yields were quantitative in baked biscuits but lower in extruded kibbles due to more dense, complex molecular structures. ASE extraction yields of 40 min or less were comparable to manual extraction yields of 24–48 h, with lower oxidation. Overall, one or two ASE extraction cycles with static times less than 20 min appeared to provide adequate lipid yields that accurately reflect lipid composition while inducing minimal modification when lipid oxidation products are the analytical endpoint.
To evaluate prognostic factors for relapse-free survival (RFS) and overall survival (OS) and to assess the impact of different postremission therapies in adult patients with core binding factor (CBF) ...acute myeloid leukemias (AML).
Individual patient data-based meta-analysis was performed on 392 adults (median age, 42 years; range, 16 to 60 years) with CBF AML (t(8;21), n = 191; inv(16), n = 201) treated between 1993 and 2002 in prospective German AML treatment trials.
RFS was 60% and 58% and OS was 65% and 74% in the t(8;21) and inv(16) groups after 3 years, respectively. For postremission therapy, intention-to-treat analysis revealed no difference between intensive chemotherapy and autologous transplantation in the t(8;21) group and between chemotherapy, autologous, and allogeneic transplantation in the inv(16) group. In the t(8;21) group, significant prognostic variables for longer RFS and OS were lower WBC and higher platelet counts; loss of the Y chromosome in male patients was prognostic for shorter OS. In the inv(16) group, trisomy 22 was a significant prognostic variable for longer RFS. For patients who experienced relapse, second complete remission rate was significantly lower in patients with t(8;21), resulting in a significantly inferior survival duration after relapse compared with patients with inv(16).
We provide novel prognostic factors for CBF AML and show that patients with t(8;21) who experience relapse have an inferior survival duration.
Mesenchymal stem cells have been shown to mediate immunomodulatory effects. They have been used in patients with steroid-refractory acute GVHD (aGVHD), but their relevance as a therapeutic agent ...targeting aGVHD has still to be defined. In this case series, we report 13 patients with steroid-refractory aGVHD who received BM-derived MSC expanded in platelet lysate-containing medium from unrelated HLA disparate donors. MSC were characterized by their morphological, phenotypical and functional properties. All tested preparations suppressed the proliferation of in vitro activated CD4+ T cells. MSC were transfused at a median dosage of 0.9 x 10(6)/kg (range 0.6-1.1). The median number of MSC applications was 2 (range 1-5). Only two patients (15%) responded and did not require any further escalation of immunosuppressive therapy. Eleven patients received additional salvage immunosuppressive therapy concomitant to further MSC transfusions, and after 28 days, five of them (45%) showed a response. Four patients (31%) are alive after a median follow-up of 257 days, including one patient who initially responded to MSC treatment. In our patient cohort, response to MSC transfusion was lower than in the series reported earlier. However, our experience supports the potential efficacy of MSC in the treatment of steroid-refractory aGVHD.
A complex aberrant karyotype consisting of multiple unrelated cytogenetic abnormalities is associated with poor prognosis in patients with acute myeloid leukemia (AML). The European Leukemia Net ...classification and the UK Medical Research Council recommendation provide prognostic categories that differ in the definition of unbalanced aberrations as well as the number of single aberrations. The aim of this study on 3526 AML patients was to redefine and validate a cutoff for karyotype complexity in AML with regard to adverse prognosis. Our study demonstrated that (1) patients with a pure hyperdiploid karyotype have an adverse risk irrespective of the number of chromosomal gains, (2) patients with translocation t(9;11)(p21∼22;q23) have an intermediate risk independent of the number of additional aberrations, (3) patients with ⩾4 abnormalities have an adverse risk per se and (4) patients with three aberrations in the absence of abnormalities of strong influence (hyperdiploid karyotype, t(9;11)(p21∼22;q23), CBF-AML, unique adverse-risk aberrations) have borderline intermediate/adverse risk with a reduced overall survival compared with patients with a normal karyotype.
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