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Human Blood (human + blood)
Kinds of Human Blood Terms modified by Human Blood Selected AbstractsCombining Fluorescent Probes and Biofunctional Magnetic Nanoparticles for Rapid Detection of Bacteria in Human Blood,ADVANCED MATERIALS, Issue 23 2006J. Gao Detection of bacteria in human blood within two hours is achieved through the use of vancomycin-functionalized FePt nanoparticles in combination with a vancomycin-conjugated fluorescent probe, as shown in the figure (B: bacteria; Van: vancomycin; FLA: fluorescein amine). This promises to be a sensitive and rapid protocol for detecting bacteria in blood products or other targets in biological samples. [source] Tirofiban and Activated Protein C Synergistically Inhibit the Instant Blood Mediated Inflammatory Reaction (IBMIR) from Allogeneic Islet Cells Exposure to Human BloodAMERICAN JOURNAL OF TRANSPLANTATION, Issue 7 2009S. Akima Instant blood mediated inflammatory reaction (IBMIR) occurs when islets are exposed to blood and manifests clinically as portal vein thrombosis and graft failure. The aim of this study was to determine the impact of recombinant human activated protein C (rhAPC) and platelet inhibition on IBMIR in order to develop a better targeted treatment for this condition. Five thousand human islet cell equivalents (IEQ) were mixed in a PVC loop system with 7 mL of ABO compatible human blood and incubated with rhAPC, either alone or in combination with tirofiban. Admixing human islets and blood caused rapid clot formation, consumption of platelets, leukocytes, fibrinogen, coagulation factors and raised d -dimers. Islets were encased in a fibrin and platelet clot heavily infiltrated with neutrophils. Tirofiban monotherapy was ineffective, whereas rhAPC monotherapy prevented IBMIR in a dose-dependent manner, preserving islet integrity while maintaining platelet and leukocyte counts, fibrinogen and coagulation factor levels, and reducing d -dimer formation. The combination of tirofiban and low-dose rhAPC inhibited IBMIR synergistically with an efficacy equal to high dose rhAPC. Tirofiban and rhAPC worked synergistically to preserve islets, suggesting that co-inhibition of the platelet and coagulation pathways' contribution to thrombin generation is required for the optimal anti-IBMIR effect. [source] Consequences Arising From Elevated Surface Temperatures on Human BloodARTIFICIAL ORGANS, Issue 9 2010Kathrin F. Hamilton Abstract Heat in blood pumps is generated by losses of the electrical motor and bearings. In the presented study the influence of tempered surfaces on bulk blood and adhesions on these surfaces was examined. Titanium alloy housing dummies were immersed in 25 mL heparinized human blood. The dummies were constantly tempered at specific temperatures (37,45°C) over 15 min. Blood samples were withdrawn for blood parameter analysis and the determination of the plasmatic coagulation cascade. The quantities of adhesion on surfaces were determined by drained weight. Blood parameters do not alter significantly up to surface temperatures of 45°C. In comparison to the control specimen, a drop in the platelet count can be observed, but is not significantly temperature dependent. The mean mass of adhesions at 41°C increased up to 66% compared to 37°C. Thus, heat generated in electrical motors and contact bearings may influence the amount of adhesions on surfaces. [source] Human monocyte CD163 expression inversely correlates with soluble CD163 plasma levelsCYTOMETRY, Issue 1 2005Bruce H. Davis Abstract Background CD163 is a monocyte/macrophage-restricted receptor involved in the clearance of hemoglobin,haptoglobin complexes and regulation of inflammatory processes. CD163 is shed from the cell surface and exists as a soluble form in plasma (sCD163). Monocyte CD163 and sCD163 are potential diagnostic tools in variety of disease states. Methods We determined the relation between plasma sCD163 levels by enzyme-linked immunosorbent assay, membrane expressions of CD163, CD64, and CD14 on blood monocytes by flow cytometry, and monocyte counts in 129 random blood samples. Results A strong inverse correlation was found between membrane CD163 expression and sCD163 levels (r = ,0.65, P < 0.001). Monocyte CD163 expression and SCD163 levels did not correlate with the monocyte absolute count. Conclusions The inverse relation between monocyte surface CD163 expression and sCD163 levels in human blood suggests that plasma sCD163 is derived from circulating monocytes, in addition to an unknown component from tissue macrophages. The lack of correlation with the absolute monocyte number suggests that such a balance is driven by the functional state of monocytes, rather than simply by numerical changes in circulating cells. We propose that further clinical evaluations of CD163 as a diagnostic parameter should include simultaneous measurements of soluble and cell-bound forms of this antigen. © 2004 Wiley-Liss, Inc. [source] Flexible Ultrathin PolyDVB/EVB Composite Membranes for the Optimization of a Whole Blood Glucose SensorELECTROANALYSIS, Issue 4 2007Kerry Bridge Abstract An ultrathin composite membrane has been developed as the outer covering barrier in a model amperometric glucose oxidase enzyme electrode. The membrane was formed by cathodic electropolymerization of divinylbenzene/ethylvinylbenzene at the surface of a gold coated polyester support membrane. Permeability coefficients were determined for O2 and glucose across membranes with a range of polymer thicknesses. Anionic interferents (such as ascorbate), were screened from the working electrode via a charge exclusion mechanism. The enzyme electrode showed an initial 10% signal drift when first exposed to whole human blood over a period of 2 hours, after which responses remained essentially stable. Whole blood patient glucose determinations yielded a correlation coefficient of r2=0.99 compared to standard hospital analyses. [source] Polydivinylbenzene/Ethylvinylbenzene Composite Membranes for the Optimization of a Whole Blood Glucose SensorELECTROANALYSIS, Issue 1 2006Kerry Bridge Abstract A novel ultra thin polydivinylbenzene/ethylvinylbenzene composite membrane has been developed for use as the outer covering barrier in a model amperometric glucose oxidase enzyme electrode. The composite membrane was formed via the cathodic electropolymerization of divinylbenzene/ethylvinylbenzene at the surface of gold sputter coated host alumina membranes, (serving solely as a mechanical support for the thin polymer film). Permeability coefficients were determined for the enzyme substrates, O2 and glucose, across composite membranes formed with a range of polymer thicknesses. Due to the highly substrate diffusion limiting nature of the composite membrane, it was found that anionic interferents present in blood (such as ascorbate), were effectively screened from the working electrode via a charge exclusion mechanism, in a manner similar to previous findings within our laboratory. The enzyme electrode showed an initial 32% signal drift when first exposed to whole human blood over a period of 2 hours, after which time enzyme electrode responses remained essentially stable. Whole blood patient glucose determinations yielded a correlation coefficient of r2=0.97 in comparison to standard hospital analyses. [source] Persistent inhibition of human natural killer cell function by ziram and pentachlorophenolENVIRONMENTAL TOXICOLOGY, Issue 4 2005Thyneice R. Taylor Abstract Ziram is a currently used agricultural fungicide. It is also used as an additive in the production of latex gloves. Because of these uses, there is a potential for human exposure to this compound. Pentachlorophenol (PCP) has been used as an insecticide, fungicide, disinfectant, and ingredient in antifouling paints. Currently, it is used as a wood preservative for power-line poles and fence posts. Measurable levels of PCP have been detected in human blood and urine. In previous studies we demonstrated that both these compounds could cause very significant inhibition of the tumor-killing function of human natural killer (NK) cells. NK lymphocytes play a central role in immune defense against viral infection and the formation of primary tumors. So interference with their function could increase the risk of tumor development. In the present study we examined the effects of exposure to ziram or PCP of brief duration (1 h) on the ability of NK cells to destroy tumor cells. NK cells were exposed to either ziram (5,0.5 ,M) or PCP (10,5 ,M) for 1 h followed by 0 h, 24 h, 48 h, or 6 days in compound-free media and then were tested for the ability to lyse as well as to bind tumor cells. A 1-h exposure to as little as 2.5 ,M ziram decreased the ability of NK cells to lyse target tumor cells, which persisted up to 6 days following exposure. The loss of lytic function for from 24 h to 6 days following exposure was accompanied by a comparable loss of NK capacity to bind tumor cells. Exposure to 10 ,M PCP for 1 h caused a progressive loss (greater than 80%) of lytic function within 6 days of exposure. In contrast to ziram, PCP exposure caused no accompanying loss of binding function. © 2005 Wiley Periodicals, Inc. Environ Toxicol 20: 418,424, 2005. [source] Amiodarone Attenuates Fluoride-induced Hyperkalemia in VitroACADEMIC EMERGENCY MEDICINE, Issue 2 2003Mark Su MD Abstract Poisoning by hydrofluoric acid or fluoride salts results in hypocalcemia, hypomagnesemia, and hyperkalemia with subsequent cardiac dysrhythmias. In previous studies, quinidine attenuated fluoride-induced hyperkalemia in vitro, and enhanced survival in animals. Like quinidine, amiodarone is a potassium channel blocker, although amiodarone is more familiar to clinicians due to its recent inclusion in advanced cardiac life support (ACLS) protocols. Objectives: This in-vitro study of human erythrocytes was designed to determine whether amiodarone could attenuate fluoride-induced hyperkalemia. Methods: Six healthy volunteers each donated 60 mL of blood on three occasions. Each specimen was divided into 12 tubes, incubated at 37°C, and oxygenated with room air. An aqueous sodium fluoride (F,) solution was added to tubes 1,9. Incremental amounts of quinidine were added to tubes 1,4 (Q1,Q4) to attain calculated concentrations of 0.73 ,g/mL, 1.45 ,g/mL, 2.9 ,g/mL, and 5.8 ,g/mL, respectively. Incremental amounts of amiodarone were added to tubes 5,8 (A1,A4) to attain calculated concentrations of 0.38 ,g/mL, 0.75 ,g/mL, 1.5 ,g/mL, and 3.0 ,g/mL, respectively. Tubes 9,12 were controls for each of F,, amiodarone, quinidine alone, and no additive, respectively. Extracellular potassium concentration ([K+]) was followed, and an objective endpoint was defined as the rise in potassium concentration at 6 hours. Results: Fluoride produced a significant change in [K+] by 6 hours in all samples. Quinidine produced a J-shaped curve in its ability to attenuate the rise in [K+], with only one concentration, Q3, demonstrating significance versus tube 9 (control). Amiodarone also demonstrated a J-shaped dose,response effect, with statistical significance at A1, A2, and A3 versus tube 9 (control). There was no significant difference among the effective concentrations (Q3, A1, A2, and A3) of both drugs. Conclusions: In this in-vitro model using human blood, amiodarone and quinidine both attenuated F, -induced hyperkalemia. Further study is indicated to determine whether amiodarone enhances survival in F, -poisoned animals. [source] Evaluation of effects of rofecoxib on platelet function in an in vitro model of thrombosis with circulating human bloodEUROPEAN JOURNAL OF CLINICAL INVESTIGATION, Issue 4 2004M. R. Hernandez Abstract Background, Cyclooxygenase (COX)-2-selective non-steroidal anti-inflammatory drugs have been used for anti-inflammatory therapy. However, it has also been described that they may increase risk of cardiovascular events. Objectives, To study the effects of COX2 inhibitor rofecoxib on platelet function using in vitro tests. Results were compared with those obtained in a parallel experiment with acetyl salicylic acid (ASA). Methods, Studies of platelet aggregation, using different agonists, were performed by a turbidimetric method. Adhesive and cohesive function of platelets were analyzed by perfusion techniques, treated blood was exposed to thrombogenic surfaces and platelet interaction was morphometrically evaluated. Results, Twenty-five µM of rofecoxib induced a prolonged lag time and a reduction in the percentage of aggregation when arachidonic acid, ADP or collagen were used as agonists. In perfusion studies with parallel chamber rofecoxib 50 µM and ASA 500 µM reduced overall platelet interaction with the collagen surface (17·4 ± 3·7, P < 0·05; vs. 32·1 ± 2·6%P < 0·05 and 17·9 ± 2·4, vs. 31·9 ± 3·24, P < 0·05, respectively). In studies performed on annular chambers, 25 µM of rofecoxib reduced platelet interaction; values of the thrombus and covered surface were 17·4 ± 4·5%; P < 0·05 and 21·1 ± 4·1%; P < 0·05, respectively, vs. 30·4 ± 7·5% and 33·5 ± 6·5 in the control. ASA did also impair thrombus formation but differences did not reach the levels of statistical significance. Moreover, rofecoxib but not ASA reduced significantly thrombus height and thrombus area (7·4 ± 0·5 µM; P < 0·005 and 96·0 ± 21·2 µM2; P < 0·05 vs. control 11·2 ± 0·9 µM and 220·0 ± 47·7µM2, respectively). Conclusion, We conclude that under our experimental conditions, rofecoxib diminished platelet aggregation induced by different agonists and inhibited platelet-mediated thrombogenesis in an in vitro model of thrombosis. [source] Polypropylene glycol is a selective binding inhibitor for LTA and other structurally related TLR2 agonistsEUROPEAN JOURNAL OF IMMUNOLOGY, Issue 3 2008Christian Draing Abstract Polypropylene glycol (PPG) is commonly added to bacterial cultures to avoid foaming. However, lipoteichoic acid (LTA) from bacteria grown with PPG lacked cytokine-inducing potency in human blood. We tested the blocking efficacy of several glycols on the cytokine response to staphylococcal LTA in human blood. PPG 1200 was the most potent inhibitor tested, shown for TNF, IL-1,, IL-6, IL-8, IL-10 and TGF-, induction, and displayed no cytotoxic effects. TNF induction by Staphylococcus aureus or by Toll-like receptor (TLR)2 agonists (di- and triacylated lipopeptides and LTA) was also inhibited by PPG 1200, but not that induced by Escherichia,coli or TLR4 agonists. In flow cytometric studies, PPG-carrying nanobeads bound more rhodamine-labeled LTA than those with glycerol. Additionally, the methyl group peak in the 1H-NMR of LTA shifted after incubation with increasing PPG 1200 concentrations. Sequential incubation of polystyrene plates with LTA, then PPG 1200 and then blood, with washing steps in between, showed that LTA-induced TNF release was inhibited. But when PPG 1200 was pre-incubated with blood that was washed before LTA was added, TNF induction was not repressed, demonstrating that PPG binds LTA and not cellular structures. In summary, PPG 1200 is a novel inhibitor of cytokine induction by TLR2 agonists, which interferes directly with the ligands. [source] CD4 T,cell activation by myelin oligodendrocyte glycoprotein is suppressed by adult but not cord blood CD25+ T,cellsEUROPEAN JOURNAL OF IMMUNOLOGY, Issue 3 2003Kajsa Wing Abstract Regulatory T,cells expressing CD25 have been shown to protect rodents from organ-specific autoimmune diseases. Similar CD25+ cells with a memory phenotype exerting suppressive function after polyclonal or allogeneic stimulation are also present in adult human blood. We demonstrate that adult human CD25+ cells regulate the response to myelin oligodendrocyte glycoprotein (MOG), as depletion of CD25+ cells increases responses of PBMC and the addition of purified CD25+ cells suppresses MOG-specific proliferation and IFN-, production of CD4+CD25, T,cells. In contrast, cord blood CD25+ cells do not inhibit responses to self antigens, and only a small subpopulation of cord CD25+ cells expresses the typical phenotype of adult regulatory T,cells (CD45RA, and GITR+) enabling suppression of polyclonal responses. We conclude that activation of self-reactive T,cells in normal healthy individuals is prevented by the presence of self-antigen-specific CD25+ regulatory T,cells and that the majority of these cells mature after birth. [source] The human basophil , a novel target of the neuropeptide alpha-melanocyte-stimulating hormoneEXPERIMENTAL DERMATOLOGY, Issue 8 2006M. Böhm There is increasing evidence that the basophil does not only play an important role in acute allergic reactions but also in the pathogenesis of chronic allergic disorders. Here we show that human basophils express melanocortin receptors (MC-Rs) and respond to alpha-melanocyte-stimulating hormone (alpha-MSH) with regulation of proallergic cytokine expression and modulation of basophil activation markers. Using primers against all known MC-R subtypes we demonstrate that the human basophil cell line KU812 expresses MC-1R. Expression of MC-1R on the surface of KU812 cells was confirmed by FACS analysis using an anti-MC-1R antibody. The MC-1R expressed by KU812 cells was functionally active as alpha-MSH induced intracellular cAMP in a dose-dependent manner. Moreover, alpha-MSH abrogated the effect of calcium ionophore A23187 on IL-4 mRNA expression in these cells. The relevance of the above findings was corroborated by showing that MC-1R surface expression is also detectable in basophils of leukocyte suspensions derived from whole human blood. Most interestingly, alpha-MSH was capable of suppressing the inductive effect of fMLP on surface expression of the basophil activation marker CD63 in leukocyte suspensions of atopic individuals. Likewise, alpha-MSH significantly blocked grass pollen-induced up-regulation of CD63 in leukocyte suspensions of patients with grass pollen allergy. Our findings highlight a novel functional dimension of alpha-MSH. In addition, MSH peptides may become a novel future therapeutic avenue in treating human allergic diseases. [source] In vitro response to Candida albicans in cultures of whole human blood from young and aged donorsFEMS IMMUNOLOGY & MEDICAL MICROBIOLOGY, Issue 2 2007Celia Murciano Abstract Invasive infections with opportunistic fungi, such as Candida albicans, have become an increasing problem in aged adults in recent years. This work investigates the influence of human ageing on C. albicans recognition by toll-like receptors (TLRs), essential components of the innate immune system, using a cohort of 96 young (15,42 years) and aged (>70 years) human volunteers. No significant differences between aged and young donors were observed on (1) cell surface TLR2, TLR6 and TLR4 expression on lymphocytes, monocytes and granulocytes, (2) production of cytokines [IL-8, IL-1,, IL-6, IL-10, tumour necrosis factor (TNF)-, and IL-12p70] and prostaglandin E2 (PGE2) by whole human blood in response to C. albicans and (3) fungicidal activity of whole blood. A statistically significant higher titre of natural anti- C. albicans antibodies was found in plasma of volunteers between 80 and 95 years old when compared with other age groups, probably as a consequence of the increased levels of serum Ig that has been described in elderly subjects. Therefore, the results indicate that the increased susceptibility to C. albicans infections in the elderly is not a consequence of defects in TLRs expression or signalling, nor of an impaired fungicidal activity of blood. [source] Combining Fluorescent Probes and Biofunctional Magnetic Nanoparticles for Rapid Detection of Bacteria in Human Blood,ADVANCED MATERIALS, Issue 23 2006J. Gao Detection of bacteria in human blood within two hours is achieved through the use of vancomycin-functionalized FePt nanoparticles in combination with a vancomycin-conjugated fluorescent probe, as shown in the figure (B: bacteria; Van: vancomycin; FLA: fluorescein amine). This promises to be a sensitive and rapid protocol for detecting bacteria in blood products or other targets in biological samples. [source] Good practice in plasma collection and fractionationISBT SCIENCE SERIES: THE INTERNATIONAL JOURNAL OF INTRACELLULAR TRANSPORT, Issue n1 2010C. Schärer The control strategy to ensure safety of blood products includes a combination of measures focusing on ensuring the quality and safety of starting material by careful donor selection and testing strategies at different levels, together with validated manufacturing processes, including steps to inactivate or remove potential contaminating agents. Using an approach based on good manufacturing practice (GMP) provides a manufacturing model that allows for a documented system of incorporating quality throughout the entire manufacturing process and describes the activities and controls needed to consistently produce products that comply with specifications and are safe for use. There are no doubts that the aim of providing safe and high-quality product to the patients should be the same for all products derived from human blood, independent of its use either as a blood component for direct transfusion or as industrially manufactured product. It would be difficult to justify whether for blood components the good practice standards and for plasma derivatives the GMP standards for manufacturing would not ensure equivalent levels of quality and safety. To ensure a high level of quality and safety of blood components and plasma derivatives, the implementation of double standards in blood establishments and fractionation industry would not be effective and should be avoided. Harmonized standards and good practices for collection and fractionation, based on the principles of GMP, should be envisaged in the whole chain of manufacturing blood components and plasma derivatives. Global initiatives to further promote the implementation of harmonized GMP for the collection in blood establishments and a stringent regulatory control are ongoing. This would further contribute to the global availability of plasma-derived medicinal products. [source] Quantitative analysis of total mitochondrial DNA: Competitive polymerase chain reaction versus real-time polymerase chain reactionJOURNAL OF BIOCHEMICAL AND MOLECULAR TOXICOLOGY, Issue 4 2004Hari K. Bhat Abstract An efficient and effective method for quantification of small amounts of nucleic acids contained within a sample specimen would be an important diagnostic tool for determining the content of mitochondrial DNA (mtDNA) in situations where the depletion thereof may be a contributing factor to the exhibited pathology phenotype. This study compares two quantification assays for calculating the total mtDNA molecule number per nanogram of total genomic DNA isolated from human blood, through the amplification of a 613-bp region on the mtDNA molecule. In one case, the mtDNA copy number was calculated by standard competitive polymerase chain reaction (PCR) technique that involves co-amplification of target DNA with various dilutions of a nonhomologous internal competitor that has the same primer binding sites as the target sequence, and subsequent determination of an equivalence point of target and competitor concentrations. In the second method, the calculation of copy number involved extrapolation from the fluorescence versus copy number standard curve generated by real-time PCR using various dilutions of the target amplicon sequence. While the mtDNA copy number was comparable using the two methods (4.92 ± 1.01 × 104 molecules/ng total genomic DNA using competitive PCR vs 4.90 ± 0.84 × 104 molecules/ng total genomic DNA using real-time PCR), both inter- and intraexperimental variance were significantly lower using the real-time PCR analysis. On the basis of reproducibility, assay complexity, and overall efficiency, including the time requirement and number of PCR reactions necessary for the analysis of a single sample, we recommend the real-time PCR quantification method described here, as its versatility and effectiveness will undoubtedly be of great use in various kinds of research related to mitochondrial DNA damage- and depletion-associated disorders. © 2004 Wiley Periodicals, Inc. J Biochem Mol Toxicol 18:180,186, 2004 Published online in Wiley InterScience (www.interscience.wiley.com). DOI 10.1002/jbt.20024 [source] Progenitor cells in vascular diseaseJOURNAL OF CELLULAR AND MOLECULAR MEDICINE, Issue 3 2005Neil Roberts Abstract Stem cell research has the potential to provide solutions to many chronic diseases via the field of regeneration therapy. In vascular biology, endothelial progenitor cells (EPCs) have been identified as contributing to angiogenesis and hence have therapeutic potential to revascularise ischaemic tissues. EPCs have also been shown to endothelialise vascular grafts and therefore may contribute to endothelial maintenance. EPC number has been shown to be reduced in patients with cardiovascular disease, leading to speculation that atherosclerosis may be caused by a consumptive loss of endothelial repair capacity. Animal experiments have shown that EPCs reendothelialise injured vessels and that this reduces neointimal formation, confirming that EPCs have an atheroprotective effect. Smooth muscle cell accumulation in the neointimal space is characteristic of many forms of atherosclerosis, however the source of these cells is now thought to be from smooth muscle progenitor cells (SMPCs) rather than the adjacent media. There is evidence for the presence of SMPCs in the adventitia of animals and that SMPCs circulate in human blood. There is also data to support SMPCs contributing to neointimal formation but their origin remains unknown. This article will review the roles of EPCs and SMPCs in the development of vascular disease by examining experimental data from in vitro studies, animal models of atherosclerosis and clinical studies. [source] From fibroblasts to iPS cells: Induced pluripotency by defined factorsJOURNAL OF CELLULAR BIOCHEMISTRY, Issue 4 2008Rui Zhao Abstract Patient-specific pluripotent cells may serve as a limitless source of transplantable tissue to treat a number of human blood and degenerative diseases without causing immune rejection. Recently, isolation of patient-specific induced pluripotent stem (iPS) cells was achieved by transducing fibroblasts with four transcription factors, Oct4, Sox2, Klf4, and c-Myc. However, the use of oncogenes and retrovirus in the current iPS cell establishment protocol raises safety concerns. To generate clinical quality iPS cells, the development of novel reprogramming methods that avoid permanent genetic modification is highly desired. The molecular mechanisms that mediate reprogramming are essentially unknown. We argue that establishment of a stable and self-sustainable ES-specific transcriptional regulatory network is essential for reprogramming. Such a system should include expression of Oct4, Sox2, Nanog and probably other pluripotenty-promoting factors from endogenous loci and establishment of a permissive epigenetic state to maintain such expression. In addition, though not yet proven experimentally, overcoming cellular senescence of fibroblasts by inactivating Rb and p53 pathways and up-regulating telomerase activity may also be required. J. Cell. Biochem. 105: 949,955, 2008. © 2008 Wiley-Liss, Inc. [source] Controlled application and removal of liposomal therapeutics: Effective elimination of pegylated liposomal doxorubicin by double-filtration plasmapheresis in vitroJOURNAL OF CLINICAL APHERESIS, Issue 2 2010Gerhard Pütz Abstract Introduction: Nanoscale particle-based drug delivery systems like long circulating liposomal doxorubicin show unique pharmacokinetic properties and improved toxicity profiles. Liposomal doxorubicin accumulates in tumor tissue due to the enhanced permeation and retention effect, but only a small fraction of a total dose reaches the tumor site. Accumulation of liposomal doxorubicin is much faster in tumor sites than in certain organs where dose limiting adverse effects occur. Finding a way to detoxify the predominant part of a given dose, circulating in the blood after accumulation is completed, will presumably reduce severe side effects during chemotherapy. Methods: Elimination properties of therapeutic used pegylated liposomal doxorubicin (Doxil®/Caelyx®) and therapeutic used double-filtration plasmapheresis systems were evaluated in vitro and in reconstituted human blood. Results: Liposomes can be filtered by appropriate membranes without leakage of doxorubicin up to a pressure of 1 bar. At higher pressures, liposomes (,85 nm) may squeeze through much smaller pores without significant leakage of doxorubicin, whereas decreasing pore size to ,8 nm leads to increased leakage of doxorubicin. With therapeutic used apheresis systems, liposomal doxorubicin can be efficiently eliminated out of buffer medium and reconstituted human blood. No leakage of doxorubicin was detected, even when liposomes were circulating for 48 h in human plasma before apheresis. Conclusions: Convenient apheresis techniques are capable of a safe and efficient elimination of therapeutic used liposomal doxorubicin in an experimental model system. J. Clin. Apheresis, 2010. © 2010 Wiley-Liss, Inc. [source] Spectrophotometric determination of leukocytes in bloodJOURNAL OF CLINICAL LABORATORY ANALYSIS, Issue 5 2002Huriye Kuzu-Karsilayan Abstract The determination of leukocyte concentration in human blood depending on the detection of oxidized o-dianisidine in acidic solution is studied. The oxidation of o-dianisidine was carried out by peroxidase enzymes found in leukocytes. The reaction was stopped by the addition of 4N H2SO4 to the solution, and a very stable, colored o-dianisidine derivative was obtained. The calibration graph was plotted with the recorded absorbance values at 400 nm assigned to the y-axis, and leukocyte counts in 1-mL blood samples to the x-axis. The equation of the calibration graph was y=0.0025x+0.0904, with a correlation coefficient of R=0.994. The coefficient of variation and P -value of the method were 4.00% and 0.05%, respectively. J. Clin. Lab. Anal. 16:233,236, 2002. © 2002 Wiley-Liss, Inc. [source] Lifestyle impacts on the aging-associated expression of biomarkers of DNA damage and telomere dysfunction in human bloodAGING CELL, Issue 4 2010Zhangfa Song Summary Cellular aging is characterized by telomere shortening, which can lead to uncapping of chromosome ends (telomere dysfunction) and activation of DNA damage responses. There is some evidence that DNA damage accumulates during human aging and that lifestyle factors contribute to the accumulation of DNA damage. Recent studies have identified a set of serum markers that are induced by telomere dysfunction and DNA damage, and these markers showed an increased expression in blood during human aging. Here, we investigated the influence of lifestyle factors (such as exercise, smoking, body mass) on the aging-associated expression of serum markers of DNA damage (CRAMP, EF-1,, stathmin, n -acetyl-glucosaminidase and chitinase) in comparison with other described markers of cellular aging (p16INK4a upregulation and telomere shortening) in human peripheral blood. The study shows that lifestyle factors have an age-independent impact on the expression level of biomarkers of DNA damage. Smoking and increased body mass indices were associated with elevated levels of biomarkers of DNA damage independent of the age of the individuals. In contrast, exercise was associated with an age-independent reduction in the expression of biomarkers of DNA damage in human blood. The expression of biomarkers of DNA damage correlated positively with p16INK4a expression and negatively with telomere length in peripheral blood T-lymphocytes. Together, these data provide experimental evidence that both aging and lifestyle impact on the accumulation of DNA damage during human aging. [source] Distribution and dynamic changes of sphingolipids in blood in response to platelet activationJOURNAL OF THROMBOSIS AND HAEMOSTASIS, Issue 12 2006F. DAHM Summary.,Background:,Sphingolipids are signaling molecules in a range of biological processes. While sphingosine-1-phosphate (S1P) is thought to be abundantly stored in platelets and released upon stimulation, knowledge about the distribution and function of other sphingolipids in blood is lacking. Objectives:,To analyze the sphingolipid content of blood components with special emphasis on dynamic changes in platelets. Methods:,Blood components from mice and humans were prepared by gradient centrifugation and analyzed by liquid chromatography,mass spectrometry. Additionally, murine platelets were activated in vitro and in vivo. Results:,Isolated non-activated platelets of mice were devoid of S1P, but instead contained dihydrosphingosine-1-phosphate (dhS1P), along with a high concentration of ceramide. Activation of platelets in vitro led to a loss of dhS1P and an increase in sphingosine, accompanied by a reduction of ceramide content. Platelet activation in vivo led to an immediate and continuous rise of dhS1P in plasma, while S1P remained stable. The sphingolipid distribution of human blood was markedly different from mice. Human platelets contained dhS1P in addition to S1P. Conclusions:,Mouse platelets contain dhS1P instead of S1P. Platelet activation causes loss of dhS1P and breakdown of ceramide, implying ceramidase activation. Release of dhS1P from activated platelets might be a novel signaling pathway. Finally, the sphingolipid composition of mouse and human blood shows large differences, which must be considered when studying sphingolipid biology. [source] Apoptosis: A review of pro-apoptotic and anti-apoptotic pathways and dysregulation in diseaseJOURNAL OF VETERINARY EMERGENCY AND CRITICAL CARE, Issue 6 2008Mauria A. O'Brien DVM Abstract Objective , To review the human and veterinary literature on the biology of apoptosis in health and disease. Data Sources , Data were examined from the human and veterinary literature identified through Pubmed and references listed in appropriate articles pertaining to apoptosis. Human Data Synthesis , The role of apoptosis in health and disease is a rapidly growing area of research in human medicine. Apoptosis has been identified as a component of human autoimmune diseases, Alzheimer's disease, cancer, and sepsis. Veterinary Data Synthesis , Research data available from the veterinary literature pertaining to apoptosis and its role in diseases of small animal species is still in its infancy. The majority of veterinary studies focus on oncologic therapy. Most of the basic science and human clinical research studies use human blood and tissue samples and murine models. The results from these studies may be applicable to small animal species. Conclusions , Apoptosis is the complex physiologic process of programmed cell death. The pathophysiology of apoptosis and disease is only now being closely evaluated in human medicine. Knowledge of the physiologic mechanisms by which tissues regulate their size and composition is leading researchers to investigate the role of apoptosis in human diseases such as cancer, autoimmune disease and sepsis. Because it is a multifaceted process, apoptosis is difficult to target or manipulate therapeutically. Future studies may reveal methods to regulate or manipulate apoptosis and improve patient outcome. [source] Measuring the arterial input function with gradient echo sequencesMAGNETIC RESONANCE IN MEDICINE, Issue 6 2003Matthias J.P. van Osch Abstract The measurement of the arterial input function by use of gradient echo sequences was investigated by in vitro and in vivo experiments. First, calibration curves representing the influence of the concentration of Gd-DTPA on both the phase and the amplitude of the MR signal were measured in human blood by means of a slow-infusion experiment. The results showed a linear increase in the phase velocity and a quadratic increase in ,R as a function of the Gd-DTPA concentration. Next, the resultant calibration curves were incorporated in a partial volume correction algorithm for the arterial input function determination. The algorithm was tested in a phantom experiment and was found to substantially improve the accuracy of the concentration measurement. Finally, the reproducibility of the arterial input function measurement was estimated in 16 patients by considering the input function of the left and the right sides as replicate measurements. This in vivo study showed that the reproducibility of the arterial input function determination using gradient echo sequences is improved by employing a partial volume correction algorithm based on the calibration curve for the contrast agent used. Magn Reson Med 49:1067,1076, 2003. © 2003 Wiley-Liss, Inc. [source] Rhesus monkey model for Leishmania major transmitted by Phlebotomus papatasi sandfly bitesMEDICAL AND VETERINARY ENTOMOLOGY, Issue 1 2001R. J. Probst Summary Leishmaniasis research needs a near-human model for investigations of natural infection processes, immunological responses and evaluation of treatments. Therefore, we developed a reproducible system using Leishmania major Yakimoff & Schokhor (Trypanosomatidae: Kinetoplastida), the cause of Old World zoonotic cutaneous leishmaniasis (ZCL), transmitted to rhesus monkeys Macaca mulatta (Zimmerman) (Primates: Cercopithecidae) by sandfly bites of experimentally infected Phlebotomus papatasi (Scopoli) (Diptera: Psychodidae). Eight monkeys of presumed Indian origin (Leishmania naïve) were exposed to bites of female sandflies that had been infected with L. major by membrane-feeding on human blood seeded with amastigotes isolated from hamster footpad lesions. Infection rates of membrane-fed sandflies averaged >,85% seven days after the infective feed, with uniformly high numbers of promastigotes in the stomodaeal valve region of the sandfly gut. Nodules and ulcerating dermal lesions developed on 7/8 monkeys 2,4 weeks post-bite and persisted for 3,7 months. Monkeys also developed satellite lesions beyond the area of sandfly bites on the head, but not on the chest. Three re-challenged monkeys developed lesions that healed faster than lesions from their primary challenges. After infection, monkeys developed delayed type hypersensitivity (DTH) responses to a panel of Leishmania skin test antigens (LSTA) and, when tested by ELISA and IFA, showed significant post-infection antibody titres which typically rose for ,170 days and then gradually receded during the next 100 days following the first challenge. After the second challenge, antibody titres spiked higher within ,50 days and receded more rapidly. In contrast, four rhesus macaques of Chinese origin developed no lesions following infected sandfly bites, although they raised antibodies and LSTA reactions, indicating subclinical infection. [source] Determination of residues of endosulfan in human blood by a negative ion chemical ionization gas chromatographic/mass spectrometric method: impact of long-term aerial spray exposurePEST MANAGEMENT SCIENCE (FORMERLY: PESTICIDE SCIENCE), Issue 3 2003Atmakuru Ramesh Abstract A new and sensitive analytical method using negative ion chemical ionization gas chromatography/mass spectrometry in selective ion monitoring (SIM) mode has been developed for the determination of residues of endosulfan in the human blood. The residues of endosulfan are extracted from whole blood samples without separating the serum by the addition of 60% sulfuric acid at 10,°C followed by partition with hexane,+,acetone (9,+,1 by volume). The total endosulfan is quantified as the sum of alpha-endosulfan, beta-endosulfan and endosulfan sulfate in SIM mode. The mass-fragment ions used for this purpose that are monitored for in SIM mode include endosulfan diol: 95, 169, 214, 313, alpha-endosulfan: 99, 242, 270, 406, beta-endosulfan: 99, 242, 270, 406, and endosulfan sulfate: 97, 353, 386. Recovery experiments were conducted at the concentration range 1.0,100,pg,ml,1. Results showed 112,98% recovery of total endosulfan from the whole blood samples. The relative standard deviation was 1.49,2.68%. The method was found to be highly sensitive in quantifying endosulfan residues down to the 0.1,pg,ml,1 level. Conversion of endosulfan to endosulfan diol was found to be less than 0.1% under the conditions used. The results were compared with published data. The applications of the analytical method for the determination of endosulfan residues in real samples was tested by analyzing 106 human blood samples collected from a population living in Padre village, Kasargode District, Kerala, India, where aerial spraying of endosulfan has been a common agricultural practice over the years. The results showed that none of the blood samples contained residues of endosulfan (alpha-endosulfan,+,beta-endosulfan,+,endosulfan sulfate) or endosulfan diol. The results were confirmed by the detection of the appropriate amounts in a number of these samples which had subsequently been spiked with endosulfan. © 2003 Society of Chemical Industry [source] Effect of conjugated linoleic acid grafting on the hemocompatibility of polyacrylonitrile membranePOLYMERS FOR ADVANCED TECHNOLOGIES, Issue 6 2006Fu-Chen Kung Abstract Polyacrylonitrile (PAN) membrane was hydrolyzed with NaOH(aq) and grafted with conjugated linoleic acid (CLA) via esterification with 1,3-propanediol. The resulting CLA grafted PAN membranes were characterized using Fourier transform infrared spectrometry (FT-IR) and X-ray photoelectronic spectroscopy (XPS). The effects of CLA grafting on the blood coagulation, platelet aggregation, and oxidative stress were evaluated using human blood. The complete blood count (CBC) and coagulation time (CT) was evaluated in vitro for hemocompatibility. After CLA grafting, the proliferation of human umbilical vein endothelial cells (HUVECs) on the membranes were improved. In addition, the production of reactive oxygen species (ROS) was measured by the chemiluminescence (CL) method to evaluate the oxidative stress. The results showed that the CLA-grafted PAN membrane could keep the CBC values more stable than unmodified PAN membrane. The CLA-grafted PAN membranes also showed longer CT. CLA-grafted PAN membrane could keep the CL counts of hydrogen peroxide and superoxide values more stable than unmodified PAN membrane. These results suggest that a CLA-grafted PAN membrane could offer protection for patients against oxidative stress and would be helpful for reducing the dosage of anticoagulant during hemodialysis. Copyright © 2006 John Wiley & Sons, Ltd. [source] Simultaneous quantitative determination of cyclosporine A and its three main metabolites (AM1, AM4N and AM9) in human blood by liquid chromatography/mass spectrometry using a rapid sample processing methodRAPID COMMUNICATIONS IN MASS SPECTROMETRY, Issue 5 2006Nozomu Koseki We have developed a sensitive and specific liquid chromatography/mass spectrometry (LC/MS) method for the simultaneous determination of cyclosporine A (CsA) and its three main metabolites (AM1, AM4N and AM9) in human blood. Following protein precipitation, supernatant was directly injected into the LC/MS system. Chromatographic separation was accomplished on a Symmetry C8 (4.6,×,75,mm, 3.5,µm) column with a linear gradient elution prior to detection by atmospheric pressure chemical ionization (APCI) MS using selected ion monitoring (SIM) in positive mode. This method can be applied to single mass equipment. The analytical range for each analyte was set at 1,2500,ng/mL using 100,µL of blood sample. The analytical method was fully validated according to FDA guidance. Intra-day mean accuracy and precision were 95.2,113.5% and 0.9,8.9%, respectively. Inter-day mean accuracy and precision were 95.8,107.0% and 1.5,10.7%, respectively. In blood all analytes were stable during three freeze/thaw cycles, for 24,h at room temperature and for 12 months at or below ,15°C. Stability was also confirmed in processed samples for 24,h at 10°C and for 6 months at 4°C in methanol. In addition, we confirmed the method could avoid matrix effects from transplant subjects' samples. This LC/MS technique provided an excellent method for simultaneous quantitative determination of CsA and its three metabolites for evaluation of their pharmacokinetic profiles. Copyright © 2006 John Wiley & Sons, Ltd. [source] Determination of clenbuterol concentration in human blood using liquid chromatography with electrospray/ion-trap tandem mass spectrometryRAPID COMMUNICATIONS IN MASS SPECTROMETRY, Issue 23 2005Ada H. Y. Yuen First page of article [source] Tirofiban and Activated Protein C Synergistically Inhibit the Instant Blood Mediated Inflammatory Reaction (IBMIR) from Allogeneic Islet Cells Exposure to Human BloodAMERICAN JOURNAL OF TRANSPLANTATION, Issue 7 2009S. Akima Instant blood mediated inflammatory reaction (IBMIR) occurs when islets are exposed to blood and manifests clinically as portal vein thrombosis and graft failure. The aim of this study was to determine the impact of recombinant human activated protein C (rhAPC) and platelet inhibition on IBMIR in order to develop a better targeted treatment for this condition. Five thousand human islet cell equivalents (IEQ) were mixed in a PVC loop system with 7 mL of ABO compatible human blood and incubated with rhAPC, either alone or in combination with tirofiban. Admixing human islets and blood caused rapid clot formation, consumption of platelets, leukocytes, fibrinogen, coagulation factors and raised d -dimers. Islets were encased in a fibrin and platelet clot heavily infiltrated with neutrophils. Tirofiban monotherapy was ineffective, whereas rhAPC monotherapy prevented IBMIR in a dose-dependent manner, preserving islet integrity while maintaining platelet and leukocyte counts, fibrinogen and coagulation factor levels, and reducing d -dimer formation. The combination of tirofiban and low-dose rhAPC inhibited IBMIR synergistically with an efficacy equal to high dose rhAPC. Tirofiban and rhAPC worked synergistically to preserve islets, suggesting that co-inhibition of the platelet and coagulation pathways' contribution to thrombin generation is required for the optimal anti-IBMIR effect. [source] | |||||||||||||||||||