WorldWideScience

Sample records for dialyzable leukocyte extracts

  1. Herpes Murine Model as a Biological Assay to Test Dialyzable Leukocyte Extracts Activity

    OpenAIRE

    Nohemí Salinas-Jazmín; Sergio Estrada-Parra; Miguel Angel Becerril-García; Alberto Yairh Limón-Flores; Said Vázquez-Leyva; Emilio Medina-Rivero; Lenin Pavón; Marco Antonio Velasco-Velázquez; Sonia Mayra Pérez-Tapia

    2015-01-01

    Human dialyzable leukocyte extracts (DLEs) are heterogeneous mixtures of low-molecular-weight peptides that are released on disruption of peripheral blood leukocytes from healthy donors. DLEs improve clinical responses in infections, allergies, cancer, and immunodeficiencies. Transferon is a human DLE that has been registered as a hemoderivate by Mexican health authorities and commercialized nationally. To develop an animal model that could be used routinely as a quality control assay for Tra...

  2. Herpes Murine Model as a Biological Assay to Test Dialyzable Leukocyte Extracts Activity

    Directory of Open Access Journals (Sweden)

    Nohemí Salinas-Jazmín

    2015-01-01

    Full Text Available Human dialyzable leukocyte extracts (DLEs are heterogeneous mixtures of low-molecular-weight peptides that are released on disruption of peripheral blood leukocytes from healthy donors. DLEs improve clinical responses in infections, allergies, cancer, and immunodeficiencies. Transferon is a human DLE that has been registered as a hemoderivate by Mexican health authorities and commercialized nationally. To develop an animal model that could be used routinely as a quality control assay for Transferon, we standardized and validated a murine model of cutaneous HSV-1 infection. Using this model, we evaluated the activity of 27 Transferon batches. All batches improved the survival of HSV-1-infected mice, wherein average survival rose from 20.9% in control mice to 59.6% in Transferon-treated mice. The activity of Transferon correlated with increased serum levels of IFN-γ and reduced IL-6 and TNF-α concentrations. Our results demonstrate that (i this mouse model of cutaneous herpes can be used to examine the activity of DLEs, such as Transferon; (ii the assay can be used as a routine test for batch release; (iii Transferon is produced with high homogeneity between batches; (iv Transferon does not have direct virucidal, cytoprotective, or antireplicative effects; and (v the protective effect of Transferon in vivo correlates with changes in serum cytokines.

  3. Herpes Murine Model as a Biological Assay to Test Dialyzable Leukocyte Extracts Activity

    Science.gov (United States)

    Salinas-Jazmín, Nohemí; Estrada-Parra, Sergio; Becerril-García, Miguel Angel; Limón-Flores, Alberto Yairh; Vázquez-Leyva, Said; Pavón, Lenin; Velasco-Velázquez, Marco Antonio; Pérez-Tapia, Sonia Mayra

    2015-01-01

    Human dialyzable leukocyte extracts (DLEs) are heterogeneous mixtures of low-molecular-weight peptides that are released on disruption of peripheral blood leukocytes from healthy donors. DLEs improve clinical responses in infections, allergies, cancer, and immunodeficiencies. Transferon is a human DLE that has been registered as a hemoderivate by Mexican health authorities and commercialized nationally. To develop an animal model that could be used routinely as a quality control assay for Transferon, we standardized and validated a murine model of cutaneous HSV-1 infection. Using this model, we evaluated the activity of 27 Transferon batches. All batches improved the survival of HSV-1-infected mice, wherein average survival rose from 20.9% in control mice to 59.6% in Transferon-treated mice. The activity of Transferon correlated with increased serum levels of IFN-γ and reduced IL-6 and TNF-α concentrations. Our results demonstrate that (i) this mouse model of cutaneous herpes can be used to examine the activity of DLEs, such as Transferon; (ii) the assay can be used as a routine test for batch release; (iii) Transferon is produced with high homogeneity between batches; (iv) Transferon does not have direct virucidal, cytoprotective, or antireplicative effects; and (v) the protective effect of Transferon in vivo correlates with changes in serum cytokines. PMID:25984538

  4. Anti-Inflammatory Effect of Dialyzable Leukocyte Extract in Autoimmune Prostatitis: Evaluation in Animal Model

    Science.gov (United States)

    Pérez-Alvarado, Carlos; Gómez, Consuelo; Reyes, Miguel; García, Mario; Pérez, Elizabeth; Pérez de la Mora, Carlos; Sanchez, Virginia

    2017-01-01

    Objective. To evaluate the anti-inflammatory properties of Dialyzable Leukocyte Extract (DLE) in a murine model of chronic prostatitis/chronic pelvic pain syndrome (CP/CPPS). Methods. Histopathological characterization, prostatein Enzyme-Linked Immunosorbent Assay, and immunohistochemical analysis for CD45, TNF-α, IFN-γ, IL-6, IL-17, and IL-4 molecules were done in prostatic Wistar rats treated with DLE, placebo, or Dexamethasone. Results. Histopathological analysis of animals induced to prostatitis showed inflammatory infiltrate, mainly constituted by leucocytes and mast cells as well as Benign Prostatic Hyperplasia. Serum prostatein concentrations were 14 times higher than those displayed by healthy animals. After DLE and Dexamethasone treatments, the inflammatory infiltrate decreased; the tissue morphology was similar to that of a normal prostate, and the prostatein decreased to the basal levels of healthy animals. DLE treatment produced a decreased expression of the cell surface marker CD45 and the proinflammatory cytokines TNF-α, IFN-γ, IL-6, and IL-17. On the other hand, the expression of anti-inflammatory cytokine IL-4 increased in both the Dexamethasone and DLE groups. Conclusion. DLE is able to modulate the inflammatory response in Experimental Autoimmune Prostatitis (EAP).

  5. In Vivo Chemoprotective Activity of Bovine Dialyzable Leukocyte Extract in Mouse Bone Marrow Cells against Damage Induced by 5-Fluorouracil

    Directory of Open Access Journals (Sweden)

    Erika Evangelina Coronado-Cerda

    2016-01-01

    Full Text Available Chemotherapy treatments induce a number of side effects, such as leukopenia neutropenia, peripheral erythropenia, and thrombocytopenia, affecting the quality of life for cancer patients. 5-Fluorouracil (5-FU is wieldy used as myeloablative model in mice. The bovine dialyzable leukocyte extract (bDLE or IMMUNEPOTENT CRP® (ICRP is an immunomodulatory compound that has antioxidants and anti-inflammatory effects. In order to investigate the chemoprotection effect of ICRP on bone marrow cells in 5-FU treated mice, total bone marrow (BM cell count, bone marrow colony forming units-granulocyte/macrophage (CFU-GM, cell cycle, immunophenotypification, ROS/superoxide and Nrf2 by flow cytometry, and histological and hematological analyses were performed. Our results demonstrated that ICRP increased BM cell count and CFU-GM number, arrested BM cells in G0/G1 phase, increased the percentage of leukocyte, granulocytic, and erythroid populations, reduced ROS/superoxide formation and Nrf2 activation, and also improved hematological levels and weight gain in 5-FU treated mice. These results suggest that ICRP has a chemoprotective effect against 5-FU in BM cells that can be used in cancer patients.

  6. In Vivo Chemoprotective Activity of Bovine Dialyzable Leukocyte Extract in Mouse Bone Marrow Cells against Damage Induced by 5-Fluorouracil

    Science.gov (United States)

    Coronado-Cerda, Erika Evangelina; Franco-Molina, Moisés Armides; Mendoza-Gamboa, Edgar; Prado-García, Heriberto; Rivera-Morales, Lydia Guadalupe; Zapata-Benavides, Pablo; Rodríguez-Salazar, María del Carmen; Caballero-Hernandez, Diana; Tamez-Guerra, Reyes Silvestre; Rodríguez-Padilla, Cristina

    2016-01-01

    Chemotherapy treatments induce a number of side effects, such as leukopenia neutropenia, peripheral erythropenia, and thrombocytopenia, affecting the quality of life for cancer patients. 5-Fluorouracil (5-FU) is wieldy used as myeloablative model in mice. The bovine dialyzable leukocyte extract (bDLE) or IMMUNEPOTENT CRP® (ICRP) is an immunomodulatory compound that has antioxidants and anti-inflammatory effects. In order to investigate the chemoprotection effect of ICRP on bone marrow cells in 5-FU treated mice, total bone marrow (BM) cell count, bone marrow colony forming units-granulocyte/macrophage (CFU-GM), cell cycle, immunophenotypification, ROS/superoxide and Nrf2 by flow cytometry, and histological and hematological analyses were performed. Our results demonstrated that ICRP increased BM cell count and CFU-GM number, arrested BM cells in G0/G1 phase, increased the percentage of leukocyte, granulocytic, and erythroid populations, reduced ROS/superoxide formation and Nrf2 activation, and also improved hematological levels and weight gain in 5-FU treated mice. These results suggest that ICRP has a chemoprotective effect against 5-FU in BM cells that can be used in cancer patients. PMID:27191003

  7. 357 Tear IL-4 is Decreased in Allergic Conjunctivitis Patients with Negative Skin Test After Dialyzable Leukocyte Extracts Treatment

    OpenAIRE

    Aguilar-Velazquez, Gustavo; Santacruz, Concepcion; Hernandez, Pedro M; Chavez, Raul; Estrada-Parra, Sergio; Perez-Tapia, Mayra; Jiménez-Martínez, Maria C.

    2012-01-01

    Background It has been suggested that sublingual immunotherapy induces immune regulation, however in patients with clinical ophthalmological diagnosis as allergic conjunctivitis with negative skin test reactivity (ACNST) this treatment is not useful. Dialyzable leukocyte extracts (DLE) have been used in atopic dermatitis and asthma. The aim of this work was to evaluate treatment of ACNST with DLE and to analyze the microenvironment provided by tear and serum cytokines in patients before and a...

  8. Early Differentiation of Human CD11c+NK Cells with γδ T Cell Activation Properties Is Promoted by Dialyzable Leukocyte Extracts

    Science.gov (United States)

    Ramírez-Ramírez, Dalia; Arriaga-Pizano, Lourdes Andrea; Mayani, Héctor; Estrada-Parra, Sergio

    2016-01-01

    Reconstitution of the hematopoietic system during immune responses and immunological and neoplastic diseases or upon transplantation depends on the emergent differentiation of hematopoietic stem/progenitor cells within the bone marrow. Although in the last decade the use of dialyzable leukocyte extracts (DLE) as supportive therapy in both infectious and malignant settings has increased, its activity on the earliest stages of human hematopoietic development remains poorly understood. Here, we have examined the ability of DLE to promote replenishment of functional lymphoid lineages from CD34+ cells. Our findings suggest that DLE increases their differentiation toward a conspicuous CD56+CD16+CD11c+ NK-like cell population endowed with properties such as IFNy production, tumor cell cytotoxicity, and the capability of inducing γδ T lymphocyte proliferation. Of note, long-term coculture controlled systems showed the bystander effect of DLE-stromal cells by providing NK progenitors with signals to overproduce this cell subset. Thus, by direct effect on progenitor cells and through activation and remodeling of the supporting hematopoietic microenvironment, DLE may contribute a robust innate immune response by promoting the emerging lymphopoiesis of functional CD11c+ NK cells in a partially TLR-related manner. Unraveling the identity and mechanisms of the involved DLE components may be fundamental to advance the NK cell-based therapy field. PMID:27847830

  9. New and cost effective cell-based assay for Dialyzed Leukocyte Extract (DLE)-induced Jurkat cells proliferation under azathioprine treatment.

    Science.gov (United States)

    Cardoso, F M; Tomkova, M; Petrovajova, D; Bubanova, M; Ragac, O; Hornakova, T

    2017-01-31

    The human Dialyzed Leukocyte Extract (DLE) is a heterogeneous mix of oligopeptides of assays exist: E-rosette test, induction of delayed type hypersensitivity in mice, leukocyte migration and IFN-γ secretion. The animal-origin materials and in vivo assays convey a considerable logistic, ethic and economic burden, meanwhile the available in vitro assays have been reported with limited reproducibility and sometimes contradictory results. Here we are reporting a new DLE biological activity cell-based assay. The A20 and Jurkat cell lines were treated with (+Aza) or without (-Aza) azathioprine, DLE (+DLE) or both (+Aza/+DLE). After 72h, the cell proliferation was analyzed by the MTT or BrdU incorporation assays. In +Aza/+DLE treated cells, we observed a significant higher proliferation, when compared with +Aza/-DLE. In the absence of Aza, cells did not present any proliferation difference between -DLE or +DLE treatments. Both assays, MTT and BrdU showed similar results, being the MTT test more cost effective and we select it for validation as DLE biological assay using Jurkat cells only. We tested three different lyophilized DLE batches and we found consistent results with acceptable assay reproducibility and linearity. The DLE capacity for rescuing Jurkat cell proliferation during +Aza treatment was consistent using different liquid and lyophilized DLE batches, presenting also consistent chromatographic profiles. Finally, DLE treatment in Jurkat cells did not result into significant IL-2 of IFN-γ secretion, and known lymphocyte proliferative drugs failed to rescue Jurkat cells viability in presence of +Aza, as +DLE treatment did in our MTT assay. In conclusion, our new cell-based MTT assay has excellent DLE biological activity consistency, robustness and is cost effective, presenting important advantages over previous DLE activity in vitro and in vivo assays.

  10. Leukocyte telomere length variation due to DNA extraction method.

    Science.gov (United States)

    Denham, Joshua; Marques, Francine Z; Charchar, Fadi J

    2014-12-04

    Telomere length is indicative of biological age. Shorter telomeres have been associated with several disease and health states. There are inconsistencies throughout the literature amongst relative telomere length measured by quantitative PCR (qPCR) and different extraction methods or kits used. We quantified whole-blood leukocyte telomere length using the telomere to single copy gene (T/S) ratio by qPCR in 20 young (18-25 yrs) men after extracting DNA using three common extraction methods: Lahiri and Nurnberger (high salt) method, PureLink Genomic DNA Mini kit (Life Technologies) and QiaAmp DNA Mini kit (Qiagen). Telomere length differences of DNA extracted from the three extraction methods was assessed by one-way analysis of variance (ANOVA). DNA purity differed between extraction methods used (P=0.01). Telomere length was impacted by the DNA extraction method used (P=0.01). Telomeres extracted using the Lahiri and Nurnberger method (mean T/S ratio: 2.43, range: 1.57-3.02) and PureLink Genomic DNA Mini Kit (mean T/S ratio: 2.57, range: 2.24-2.80) did not differ (P=0.13). Likewise, QiaAmp and Purelink-extracted telomeres were not statistically different (P=0.14). The Lahiri-extracted telomeres, however, were significantly shorter than those extracted using the QiaAmp DNA Mini Kit (mean T/S ratio: 2.71, range: 2.32-3.02; P=0.003). DNA purity was associated with telomere length. There are discrepancies between the length of leukocyte telomeres extracted from the same individuals according to the DNA extraction method used. DNA purity could be responsible for the discrepancy in telomere length but this will require validation studies. We recommend using the same DNA extraction kit when quantifying leukocyte telomere length by qPCR or when comparing different cohorts to avoid erroneous associations between telomere length and traits of interest.

  11. Leukocyte-stimulating effect and phytochemical screening of Trichilia hirta extracts.

    Science.gov (United States)

    Sosa, Edgar Hernandez; Castejón, Yulianis Martín; Duharte, Alexander Batista; Portuondo, Deivys; Tamayo, Vivian; Quevedo, Humberto J Morris; Manrique, Clara Esther Martínez

    2011-09-01

    Trichilia hirta (Family Meliaceae) is a tree traditionally used in the folk medicine of Cuba to treat asthma, cancer, and ulcers. The objective of this study was to determine the phytochemical composition of ethanol extracts obtained from leaves, roots, and stem bark and to evaluate the leukocyte-stimulating effect of T. hirta root extracts on BALB/c mice. The chemical composition of the extracts was determined by phytochemical screening. Saponins, tannins, flavonoids, and coumarins were detected in extracts of T. hirta. The leukocyte-stimulating effect was evaluated by oral application of ethanol extracts (81.8 and 976 mg/kg) in BALB/c mice for 7 days. The application of 976 mg of extract/kg increased the total leukocyte count up to 15-33%; this effect was significant for neutrophil counts compared with control animals (Pphytochemicals reported as immunostimulants. The administration of these extracts to BALB/c mice indicated that ethanol extract could exhibit leukocyte-stimulating properties and makes it a promising alternative for the development of an immunoprotective agent.

  12. Oxygen-independent killing of Bacteroides fragilis by granule extracts from human polymorphonuclear leukocytes.

    Science.gov (United States)

    Wetherall, B L; Pruul, H; McDonald, P J

    1984-03-01

    Granule proteins from human neutrophils were prepared by extraction with acetate, and their antibacterial activity against Bacteroides fragilis was determined. Activity was highly dependent on pH; greatest killing occurred at the most acid pH tested (pH 5.0). Optimum activity was observed at physiological ionic strength and low bacterial numbers. Killing was inhibited by incubation temperatures of less than 37 degrees C. Eight times more extract was required to kill 50% of stationary-phase bacteria, compared with those growing in logarithmic phase. The antibacterial effect of granule extract was destroyed by boiling, but some activity was retained after heating to 56 degrees C and 80 degrees C. Granule extract activity was tested under conditions in which oxygen-dependent antibacterial systems were inhibited. The rate and extent of killing was not affected by anaerobiosis, sodium azide, or cysteine hydrochloride. These results suggest that the activity of granule extract is independent of oxidative antibacterial systems, and therefore, under conditions that occur in anaerobic infections, potent leukocyte granule-associated mechanisms exist for the destruction of B. fragilis.

  13. The Peroxidation of Leukocytes Index Ratio Reveals the Prooxidant Effect of Green Tea Extract.

    Science.gov (United States)

    Peluso, Ilaria; Manafikhi, Husseen; Raguzzini, Anna; Longhitano, Yaroslava; Reggi, Raffaella; Zanza, Christian; Palmery, Maura

    2016-01-01

    Despite tea increased plasma nonenzymatic antioxidant capacity, the European Food Safety Administration (EFSA) denied claims related to tea and its protection from oxidative damage. Furthermore, the Supplement Information Expert Committee (DSI EC) expressed some doubts on the safety of green tea extract (GTE). We performed a pilot study in order to evaluate the effect of a single dose of two capsules of a GTE supplement (200 mg × 2) on the peroxidation of leukocytes index ratio (PLIR) in relation to uric acid (UA) and ferric reducing antioxidant potential (FRAP), as well as the sample size to reach statistical significance. GTE induced a prooxidant effect on leukocytes, whereas FRAP did not change, in agreement with the EFSA and the DSI EC conclusions. Besides, our results confirm the primary role of UA in the antioxidant defences. The ratio based calculation of the PLIR reduced the sample size to reach statistical significance, compared to the resistance to an exogenous oxidative stress and to the functional capacity of oxidative burst. Therefore, PLIR could be a sensitive marker of redox status.

  14. The Peroxidation of Leukocytes Index Ratio Reveals the Prooxidant Effect of Green Tea Extract

    Directory of Open Access Journals (Sweden)

    Ilaria Peluso

    2016-01-01

    Full Text Available Despite tea increased plasma nonenzymatic antioxidant capacity, the European Food Safety Administration (EFSA denied claims related to tea and its protection from oxidative damage. Furthermore, the Supplement Information Expert Committee (DSI EC expressed some doubts on the safety of green tea extract (GTE. We performed a pilot study in order to evaluate the effect of a single dose of two capsules of a GTE supplement (200 mg × 2 on the peroxidation of leukocytes index ratio (PLIR in relation to uric acid (UA and ferric reducing antioxidant potential (FRAP, as well as the sample size to reach statistical significance. GTE induced a prooxidant effect on leukocytes, whereas FRAP did not change, in agreement with the EFSA and the DSI EC conclusions. Besides, our results confirm the primary role of UA in the antioxidant defences. The ratio based calculation of the PLIR reduced the sample size to reach statistical significance, compared to the resistance to an exogenous oxidative stress and to the functional capacity of oxidative burst. Therefore, PLIR could be a sensitive marker of redox status.

  15. Morinda citrifolia Linn leaf extract possesses antioxidant activities and reduces nociceptive behavior and leukocyte migration.

    Science.gov (United States)

    Serafini, Mairim Russo; Santos, Rodrigo Correia; Guimarães, Adriana Gibara; Dos Santos, João Paulo Almeida; da Conceicão Santos, Alan Diego; Alves, Izabel Almeida; Gelain, Daniel Pens; de Lima Nogueira, Paulo Cesar; Quintans-Júnior, Lucindo José; Bonjardim, Leonardo Rigoldi; de Souza Araújo, Adriano Antunes

    2011-10-01

    Herbal drugs have been used since ancient times to treat a wide range of diseases. Morinda citrifolia Linn (popularly known as "Noni") has been used in folk medicine by Polynesians for over 2,000 years. It is reported to have a broad range of therapeutic effects, including effects against headache, fever, arthritis, gingivitis, respiratory disorders, infections, tuberculosis, and diabetes. The aim of this study was to investigate the antioxidant, anti-inflammatory, antinociceptive, and antibacterial properties of the aqueous extract from M. citrifolia leaves (AEMC). Antioxidant activity was observed against lipid peroxidation, nitric oxide, and hydroxyl radicals. The antinociceptive effect of AEMC was observed in the acetic acid-induced writhing test at the higher dose. Moreover, AEMC significantly reduced the leukocyte migration in doses of 200 and 400 mg/kg and showed mild antibacterial activity. Together, the results suggest that properties of M. citrifolia leaf extract should be explored further in order to achieve newer tools for managing painful and inflammation conditions, including those related to oxidant states.

  16. Effects of Methanolic Jatropha multifida L. Extract in Wound Healing Assessed by the Total Number of PMN Leukocytes and Fibroblasts

    Directory of Open Access Journals (Sweden)

    Juniarti

    2012-12-01

    Full Text Available Objective: The aim of this study was to evaluate the effects of methanol extract of Jatropha multifida leaves on the wound healing process and to investigate the wound healing activity based on reduced numbers of PMN (polymorpho nuclear leukocytes and increased numbers of fibroblasts. Method: methanol extract of dried leaves of Jatropha multifida was used in the wound healing activity studies. The study subjects were 36 white male Sprague Dawlay rats aged 2 months with 150-200 gram body weight. The subjects were divided into 4 groups and experimentally injured: Group I (negative control underwent injury without subsequent treatment; group II (positive control received topical treatment with Bethasone-N after injury; group III (solvent control was treated with 70% methanol; group IV (treatment group was treated with 10 mg methanol extract of Jatropha multifida Each group consisted of 3 rats, which were decapitated on days 3, 6, and 13 after the start of treatment. Histological preparation was stained with hematoxyline-eosin (HE and was continuously examined by counting the numbers of PMN leukocytes and fibroblasts as indicators of wound healing on days 3, 6, and 13 of treatment. The study showed lower numbers of PMN leukocytes in subjects treated with the extract of Jatropha multifida as compared to the other groups. The numbers of fibroblasts were significantly higher on days 6 and 13 of treatment. In conclusion, the treatment of injuries with methanol extract of leaves from Jatropha multifida provided better results compared to the other groups in our study.

  17. Iron dialyzability from hospital duplicate meals: daily intake.

    Science.gov (United States)

    Velasco-Reynold, Carlos; Navarro-Alarcon, Miguel; Lopez-Ga de la Serrana, Herminia; Perez-Valero, Vidal; Lopez-Martinez, María C

    2009-09-01

    Both total and dialyzable iron levels and corresponding dialyzability were determined in 108 duplicate meals during 36 consecutive days. Total mean iron fraction of 5.90 +/- 4.97 mg was found in the meals. The iron supplied by the meals is directly and significantly (p < 0.05) correlated with macromicronutrient content (carbohydrates, fiber, and protein). The mean iron dialyzability (4.81 +/- 3.25%) was low and not significantly different among the three primary meals (breakfast, lunch, and dinner). Significant interactions of several minerals on iron levels were found (p < 0.05). Iron dialyzability was only statistically influenced by zinc dialyzability in meals (p < 0.05). The dialyzed iron fraction present in meals was significantly correlated with protein and ascorbic acid levels (p < 0.01). The mean iron daily dietary intake was 17.7 +/- 6.91 mg. The hospital meals provided enough iron. Foods of animal origin are primary sources of iron in diet.

  18. Assessment of dialyzer surface in online hemodiafiltration; objective choice of dialyzer surface area

    Directory of Open Access Journals (Sweden)

    Francisco Maduell

    2015-05-01

    Conclusion: The increase in 40% and 80% of dialyzer surface area entails an increase in convective volume of 6 and 16% respectively, showing minimal differences both in convective volume and clearance capacity when UFC was greater than 45 mL/h/mmHg. It is advisable to optimise dialyser efficiency to the smallest surface area possible, adjusting treatment prescription.

  19. Effect of Aloe vera extract on the improvement of the respiratory activity of leukocytes of matrinxã during the transport stress

    Directory of Open Access Journals (Sweden)

    Fábio Sabbadin Zanuzzo

    2012-10-01

    Full Text Available This study evaluated the effect of extract of Aloe vera in the transport water of matrinxã (Brycon amazonicus fish on stress response and leukocyte respiratory activity. Fish was transported for 4 h in water containing Aloe at levels 0; 0.02; 0.2 and 2 mg/L, and sampled before transport 2, 4, 24 and 96 h after for determination of plasma glucose and respiratory activity of leukocytes. An additional in vitro assay was conducted with another fish species, pacu (Piaractus mesopotamicus, to test the respiratory burst of leukocytes exposed to Aloe extract (0.0, phosphate-buffered saline (PBS only at 0.1, 0.2, 0.5 and 1 mg/L. Plasma glucose increased after 2 and 4 h of transport and returned to control levels within 24 h, but the addition of Aloe in the transport water did not affect the level of blood glucose. However, at 2 h of transport, Aloe enhanced the respiratory activity of leukocytes in a dose-dependent way. The highest value of respiratory burst activity of leukocytes was observed in the fish transported in water containing Aloe at 2 mg/L. The enhancing effect of the plant extract on the production of oxygen radicals was confirmed in vitro in leukocytes of pacu incubated in Aloe at concentrations 0.1 and 0.2 mg/L. The results suggest that Aloe vera is a modulator of the immune system in fish improving the innate immune response tested.

  20. Differential Effects of Tea Extracts on Growth and Cytokine Production by Normal and Leukemic Human Leukocytes

    Directory of Open Access Journals (Sweden)

    Diana Bayer

    2012-04-01

    Full Text Available Background: Tea is one of the world’s most highly consumed beverages, second only to water. It is affordable and abundant and thus has great potential for improving health of those in both developed and developing areas. Green, oolong, and black teas differ in the extent of fermentation and types of bioactive polyphenols produced. Green tea and its major polyphenol decrease growth of some cancer cells and effect production of immune system cytokines. This study compares the effects of different types of tea extracts on viability and cytokine production by normal and leukemic human T lymphocytes. Generation of the toxic reactive oxygen species H2O2 by extracts was also examined.Methods: The Jurkat T lymphoblastic leukemia cells and mitogen-stimulated normal human peripheral blood mononuclear cells were used in this study. Cell viability was determined by (3-4,5-dimethylthiamizol-2-yl-diphenyltetrazolium bromide assay and production of interleukin-2 by Enzyme-Linked ImmunoSorbent Assay. Levels of H2O2 generated by tea extracts were determined using the xylenol-orange method.Results: We found that green, oolong, and black tea extracts differentially effect the growth and viability of T lymphoblastic leukemia cells and normal peripheral blood mononuclear cells, substantially decreasing both growth and viability of leukemic T lymphocytes and having much lesser effects on their normal counterparts. Tea extracts also had differential effects on the production of the T lymphocyte growth factor interleukin-2, significantly decreasing production by leukemic cells while having only minor effects on normal cells. All three extracts induced H2O2 generation, with green and oolong tea extracts having the greatest effect. Leukemic cells were much more susceptible to growth inhibition and killing by H2O2 than normal lymphocytes.Functional Foods in Health and Disease 2012, 2(4:72-85 Conclusions: The three tea extracts studied altered leukemic T lymphocyte

  1. Oxygen-independent killing of Bacteroides fragilis by granule extracts from human polymorphonuclear leukocytes.

    OpenAIRE

    Wetherall, B L; Pruul, H; McDonald, P J

    1984-01-01

    Granule proteins from human neutrophils were prepared by extraction with acetate, and their antibacterial activity against Bacteroides fragilis was determined. Activity was highly dependent on pH; greatest killing occurred at the most acid pH tested (pH 5.0). Optimum activity was observed at physiological ionic strength and low bacterial numbers. Killing was inhibited by incubation temperatures of less than 37 degrees C. Eight times more extract was required to kill 50% of stationary-phase ba...

  2. Dialyzer Reuse Impact on Dialyzer Efficiency, Patient Morbidity and Mortality and Cost Effectiveness

    Directory of Open Access Journals (Sweden)

    Mitwalli Ahmed

    2001-01-01

    Full Text Available Since the introduction of dialyzer reuse more than three decades ago, several studies have reported its safety, efficacy and cost effectiveness. Reuse of hemodialyzer was prospectively studied in ten chronic hemodialysis (HD patients recruited from the renal unit, the King Khalid University Hospital, Riyadh, Saudi Arabia, for three months. During the study period, 66 dialyzers were used for 408 sessions of HD, with a mean reuse of 6.2 ± 5.3 episodes per dialyser, the mean of maximum reuse episodes being 13.7 ± 8.0. The urea reduction ratio was maintained between 73 ± 5% at baseline to 71.2 ± 9.03% (p=0.53 at the maximum reuse. Similarly phosphate reduction with each HD session was maintained; mean decrease in phosphate levels was 0.67 mmol/L. Significant increase in heparin requirement was noted; however, the risk of bleeding was not increased. Hematocrit levels increased from 30.4 ± 4.1% to 33.2 ± 3.6% at the end of the study (p=0.6. Albumin leak in dialysate decreased with each reuse; baseline 8.27 ± 7.93 mg/L to 2.8 ± 0.4 mg/L at maximum reuse (p=0.04. Serum albumin levels remained stable. No short-term adverse effects on patients′ morbidity and mortality were noted. Total cost savings of 53% was achieved with the reuse of dialyzers, excluding capital equipment used for preparation for reuse. In conclusion, dialyzer reuse seems to be safe and may provide an economical and efficient dialysis. Studies involving larger number of patients is required to validate this observation.

  3. Dialyzer reuse impact on dialyzer efficiency, patient morbidity and mortality and cost effectiveness.

    Science.gov (United States)

    Mitwalli, A H; Abed, J; Tarif, N; Alam, A; Al-Wakeel, J S; Abu-Aisha, H; Memon, N; Sulaimani, F; Ternate, B; Mensah, M O

    2001-01-01

    Since the introduction of dialyzer reuse more than three decades ago, several studies have reported its safety, efficacy and cost effectiveness. Reuse of hemodialyzer was prospectively studied in ten chronic hemodialysis (HD) patients recruited from the renal unit, the King Khalid University Hospital, Riyadh, Saudi Arabia, for three months. During the study period, 66 dialyzers were used for 408 sessions of HD, with a mean reuse of 6.2 +/- 5.3 episodes per dialyser, the mean of maximum reuse episodes being 13.7 +/- 8.0. The urea reduction ratio was maintained between 73 +/- 5% at baseline to 71.2 +/- 9.03% (p=0.53) at the maximum reuse. Similarly phosphate reduction with each HD session was maintained; mean decrease in phosphate levels was 0.67 mmol/L. Significant increase in heparin requirement was noted; however, the risk of bleeding was not increased. Hematocrit levels increased from 30.4 +/- 4.1% to 33.2 +/- 3.6% at the end of the study (p=0.6). Albumin leak in dialysate decreased with each reuse; baseline 8.27 +/- 7.93 mg/L to 2.8 +/- 0.4 mg/L at maximum reuse (p=0.04). Serum albumin levels remained stable. No short-term adverse effects on patients' morbidity and mortality were noted. Total cost savings of 53% was achieved with the reuse of dialyzers, excluding capital equipment used for preparation for reuse. In conclusion, dialyzer reuse seems to be safe and may provide an economical and efficient dialysis. Studies involving larger number of patients is required to validate this observation.

  4. Effects of Hypericum perforatum extract on IgG titer, leukocytes subset and spleen index in rats

    Directory of Open Access Journals (Sweden)

    Tahereh Aghili

    2014-11-01

    Full Text Available Objectives: Hypericum perforatum L. is a medicinal plant containing many polyphenolic compounds, ‎including flavonoids and phenolic acids with antidepressant and anti-inflammatory properties. ‎This study was investigated the effects of Hypericum perforatum extract (HPE on immunity, ‎body weight (BW, and spleen index (SI in rats.‎ Materials and Methods: A total of 24 Wistar male rats were randomly received 4 different doses (6 rats each of HPE ‎‎(0, 100, 200 and 400 mg/kg BW intraperitoneally for 14 days using a completely ‎randomized design. On days 1 and 7, rats were received 0.5 ml SRBC (10% injection. Blood ‎samples were collected on day 14 to evaluate IgG titer and leukocyte count. On days 1, 7 and ‎‎14, the BW and on day 14 spleen were weighted for SI. ‎ Results: The IgG titer increased with higher doses of HPE. The HPE increased number of ‎lymphocytes at 200 mg but decreased at 400 mg, number of neutrophils decreased at 200 mg ‎but increased at 400 mg, and number of monocytes increased at 100 mg and 200 mg but ‎decreased at 400 mg (p

  5. Internal filtration in dialyzers with different membrane permeabilities.

    Science.gov (United States)

    Sato, Yuichi; Kimura, Kenjiro; Chikaraishi, Tatsuya

    2010-07-01

    Over the last decade, hemodialysis with enhanced internal filtration (IF) has been investigated as an alternative to conventional dialysis. Several factors affect IF, including the geometry and permeability of hollow-fiber dialyzers. Although various studies have been performed, the association between IF and membrane permeability has not been fully examined because of the difficulty in measuring IF. Therefore, in this study, we set up an experimental circuit and attempted to directly measure IF as well as membrane permeability in five dialyzers. In the circuit, we placed two dialyzers of the same type in series, and a special sampling port between them, thereby making it possible to determine IF by measuring the extent to which blood was concentrated between the two dialyzers. We showed that a significant amount of IF occurred in this tandem-dialyzer circuit, ranging from 23.5 to 100 ml/min, which increased linearly with increasing membrane permeability. We also showed that membrane permeability was reduced in the first dialyzer to a greater extent than in the second one after four hours of circulation, suggesting that filtration caused substantial membrane fouling. In this study we practically demonstrated that membrane permeability is highly relevant to the phenomenon of IF.

  6. The past, present and future of the dialyzer.

    Science.gov (United States)

    Mineshima, Michio

    2015-01-01

    Several types of dialysis membranes have been developed in the history of hemodialysis therapy. Regenerated cellulose had been widely used for a long time, since the beginning of dialysis therapy. Regenerated cellulose is strongly hydrophilic, which enables lower membrane thickness and miniaturization of the dialyzer. The cellulose triacetate membrane has greater performance because of the lower thickness of the membrane and its lack of swelling due to high hydrophobicity. Many types of synthetic membranes, such as polysulfone, polyethersulfone and polyester polymer alloy membranes, have asymmetrical structures. Dialyzers with these membranes show higher capacities for water and solute transport because the actual membrane thickness, which is related to the water and solute transfer resistance, is quite small compared to that in membranes with homogeneous structures. The development of highly biocompatible membranes will be required in the future so as to prevent the development of adverse reactions and related complications. The performance of a dialyzer depends not only on the membrane permeability but also on the flow conditions of the blood and dialysate. Many types of dialyzers with high-performance membranes have been developed as a result of advances in membrane and device technologies. Recently, many types of high-flux dialyzers with high-performance membranes with a high internal filtration (IF)/backfiltration (BF) flow rate have been introduced. IF-enhanced hemodialysis using an enhanced IF dialyzer seems to be more convenient than hemodiafiltration therapy because it requires no additional equipment, such as a roller pump. In the near future, dialyzers should be developed with high capacities for the removal of low-molecular-weight proteins (LMWPs) related to complications and with low capacities for the loss of albumin and amino acid. Dialyzers with a sharp cut-off membrane between LMWPs and albumin and dialyzers with a special function, i.e., an

  7. Dialyzer reuse: justified cost saving for south Asian region.

    Science.gov (United States)

    Dhrolia, Murtaza F; Nasir, Kiran; Imtiaz, Salman; Ahmad, Aasim

    2014-08-01

    In spite of controversies, dialyzer reuse has remained an integral part of hemodialysis because of lower cost, good overall safety record, and improved membrane biocompatibility. Reuse declined in developed countries from the beginning of this century because of mass production of hemodialyzers at favourable price with better biocompatible membrane. Abandoning dialyzer reuse became challenging in South Asian region, where more than 40% of the population live below the International Poverty Line of $1.25 per day, less than 10% of end stage renal disease patients receive renal replacement therapy, and upto 70% of those starting dialysis stop treatment due to cost within the first 3 months. Dialyzer reuse is an efficient cost-saving method that allows the use of more efficient and expensive biocompatible synthetic membranes, thereby providing high-quality dialysis to individuals living in countries with limited medical resources without compromising the safety or effectiveness of the treatment.

  8. Dialyzer Reuse and Outcomes of High Flux Dialysis.

    Science.gov (United States)

    Argyropoulos, Christos; Roumelioti, Maria-Eleni; Sattar, Abdus; Kellum, John A; Weissfeld, Lisa; Unruh, Mark L

    2015-01-01

    The bulk of randomized trial evidence for the expanding use of High Flux (HF) hemodialysis worldwide comes from two randomized controlled trials, one of which (HEMODIALYSIS, HEMO) allowed, while the other (Membrane Outcomes Permeability, MPO) excluded, the reuse of membranes. It is not known whether dialyzer reuse has a differential impact on outcomes with HF vs low flyx (LF) dialyzers. Proportional Hazards Models and Joint Models for longitudinal measures and survival outcomes were used in HEMO to analyze the relationship between β2-microglobulin (β2M) concentration, flux, and reuse. Meta-analysis and regression techniques were used to synthesize the evidence for HF dialysis from HEMO and MPO. In HEMO, minimally reused (dialysis and the reuse process on dialyzers may suggest novel biomarkers for uremic toxicity and may accelerate membrane technology innovations that will improve patient outcomes.

  9. Dialyzer Reuse and Outcomes of High Flux Dialysis.

    Directory of Open Access Journals (Sweden)

    Christos Argyropoulos

    Full Text Available The bulk of randomized trial evidence for the expanding use of High Flux (HF hemodialysis worldwide comes from two randomized controlled trials, one of which (HEMODIALYSIS, HEMO allowed, while the other (Membrane Outcomes Permeability, MPO excluded, the reuse of membranes. It is not known whether dialyzer reuse has a differential impact on outcomes with HF vs low flyx (LF dialyzers.Proportional Hazards Models and Joint Models for longitudinal measures and survival outcomes were used in HEMO to analyze the relationship between β2-microglobulin (β2M concentration, flux, and reuse. Meta-analysis and regression techniques were used to synthesize the evidence for HF dialysis from HEMO and MPO.In HEMO, minimally reused (< 6 times HF dialyzers were associated with a hazard ratio (HR of 0.67 (95% confidence interval, 95%CI: 0.48-0.92, p = 0.015, 0.64 (95%CI: 0.44 - 0.95, p = 0.03, 0.61 (95%CI: 0.41 - 0.90, p = 0.012, 0.53 (95%CI: 0.28 - 1.02, p = 0.057 relative to minimally reused LF ones for all cause, cardiovascular, cardiac and infectious mortality respectively. These relationships reversed for extensively reused membranes (p for interaction between reuse and flux < 0.001, p = 0.005 for death from all cause and cardiovascular causes, while similar trends were noted for cardiac and infectious mortality (p of interaction between reuse and flux of 0.10 and 0.08 respectively. Reduction of β2M explained only 1/3 of the effect of minimally reused HF dialyzers on all cause mortality, while non-β2M related factors explained the apparent attenuation of the benefit with more extensively reused dialyzers. Meta-regression of HEMO and MPO estimated an adjusted HR of 0.63 (95% CI: 0.51-0.78 for non-reused HF dialyzers compared with non-reused LF membranes.This secondary analysis and synthesis of two large hemodialysis trials supports the widespread use of HF dialyzers in clinical hemodialysis over the last decade. A mechanistic understanding of the effects of

  10. Is it easier to dialyze women than men?

    Science.gov (United States)

    Maduell, F; Caridad, A; Sigüenza, F; Miralles, F; Serrato, F

    1994-01-01

    Since the urea kinetic model is used as an objective method for monitoring dialysis, it has been possible to shorten reasonably its duration. From the usual practice, we have observed that it is easier to reduce the dialysis time in women as compared to men. The purpose of this study is to find out the reason for such observation and to corroborate it. Fourty-two patients, 25 males and 17 females on 3-weekly dialysis (3 h/session) were studied. All patients were dialyzed under the same dialysis characteristics: 3-hour sessions, blood flow 350 ml/min, ultrafiltration 1.5 liters/h, 1.5-m2 cuprophane membrane and bicarbonate buffer. The dialysate was collected in a graduated tank. Urea concentration in plasma and in the dialysate was measured. Then, the urea distribution volume, dialyzer clearance, the KT/V index and protein catabolic rate were calculated. The KT/V value was higher in women with respect to men, 1,017 +/- 0.10 versus 0.82 +/- 0.14 (p < 0.001). The urea distribution volume value was significantly higher in men as compared to women, 60.04 +/- 6.6 versus 51.48 +/- 5.88% (p < 0.001). There were no significant differences in dialyzer clearance, protein catabolic rate or body weight. In conclusion, under identical dialysis conditions, it is easier to dialyze women than men, because women's urea distribution volume is lower than men's.

  11. Density-Gradient Mediated Band Extraction of Leukocytes from Whole Blood Using Centrifugo-Pneumatic Siphon Valving on Centrifugal Microfluidic Discs.

    Directory of Open Access Journals (Sweden)

    David J Kinahan

    Full Text Available Here we present retrieval of Peripheral Blood Mononuclear Cells by density-gradient medium based centrifugation for subsequent analysis of the leukocytes on an integrated microfluidic "Lab-on-a-Disc" cartridge. Isolation of white blood cells constitutes a critical sample preparation step for many bioassays. Centrifugo-pneumatic siphon valves are particularly suited for blood processing as they function without need of surface treatment and are 'low-pass', i.e., holding at high centrifugation speeds and opening upon reduction of the spin rate. Both 'hydrostatically' and 'hydrodynamically' triggered centrifugo-pneumatic siphon valving schemes are presented. Firstly, the geometry of the pneumatic chamber of hydrostatically primed centrifugo-pneumatic siphon valves is optimised to enable smooth and uniform layering of blood on top of the density-gradient medium; this feature proves to be key for efficient Peripheral Blood Mononuclear Cell extraction. A theoretical analysis of hydrostatically primed valves is also presented which determines the optimum priming pressure for the individual valves. Next, 'dual siphon' configurations for both hydrostatically and hydrodynamically primed centrifugo-pneumatic siphon valves are introduced; here plasma and Peripheral Blood Mononuclear Cells are extracted through a distinct siphon valve. This work represents a first step towards enabling on disc multi-parameter analysis. Finally, the efficiency of Peripheral Blood Mononuclear Cells extraction in these structures is characterised using a simplified design. A microfluidic mechanism, which we termed phase switching, is identified which affects the efficiency of Peripheral Blood Mononuclear Cell extraction.

  12. Fluid and solute transfer characteristics in a dialyzer with a high-performance membrane.

    Science.gov (United States)

    Mineshima, Michio

    2011-01-01

    To date, many types of high-flux dialyzers with high-performance membranes have been developed. They seem to have a higher internal filtration flow rate (Q(IF)) because of a higher ultrafiltration coefficient. In the health reimbursement system in Japan, commercially available dialyzers are categorized into five types based on their β(2)-microglobulin clearance (K(BMG)) values. Although the K(BMG) value was not correlated strongly with the Q(IF) value for 26 types of commercially available dialyzers, almost all the dialyzers with a higher Q(IF) value had a higher KBMG value. These dialyzers seem to have a higher convective transport because of internal filtration in addition to diffusive transport. We measured the blood flow velocity in a cross-sectional plane of the dialyzer using pulse Doppler ultra-sonography to evaluate QIF. It is a useful method for the bedside monitoring because it is noninvasive to the patient and produces reliable data with a higher reproducibility. On the other hand, membrane fouling occurs more easily in higher Q(IF) dialyzers, compared with conventional dialyzers, because of the higher degree of membrane fouling. Internal filtration-enhanced hemodialysis (IFEHD) using these dialyzers, therefore, has the advantage of increasing solute removal efficiency by enhancing convective transport and the simultaneous disadvantage of decreasing solute removal efficiency by causing membrane fouling. Thus, IFEHD treatment should be performed using a dialyzer with a high-performance membrane to ensure that the advantage is superior to the disadvantage.

  13. Anti-inflammatory effect of hydromethanolic extract from Marrubium vulgare Lamiaceae on leukocytes oxidative metabolism: An in vitro and in vivo studies

    Directory of Open Access Journals (Sweden)

    Ahmed El Abbouyi

    2013-01-01

    Full Text Available Background: Marrubium vulgare Lamiaceae, known as white horehound, is frequently used in folk medicine to treat many illnesses from ancient times. Aims: The present study is conducted to evaluate the anti-inflammatory activity of hydromethanolic extract of the aerial part of M. vulgare. Materials and Methods: The oxygen consumption and the production of superoxide anions, by rat pleural polymorphonuclear leukocytes stimulated by opsonized zymosan, were performed. PMNs were collected after induction of an acute inflammatory reaction by injection, in the rat pleural cavity, of suspension of calcium pyrophosphate crystals. Plant extract was tested at a concentration range 10-100 mg/ml and 100-400 mg/kg/day for in vitro and in vivo assays respectively. Statistical analysis: All values are expressed as mean ΁ standard error. The significance of differences between control and treated samples in vitro was analysed using the Student′s t-test. For in vivo studies, data were analysed using one-way analysis of variance. Differences with P ≤ 0.05 were considered to be statistically significant. Results: Hydromethanolic extract of M. vulgare showed, in vitro a significant anti-inflammatory effect in a dose-dependent manner. In vivo, only the high amounts of the plant extract (300 and 400 mg/kg/day exhibited a significant anti-inflammatory effect. The plant extract contains a high amount of polyphenols. Conclusion: Hydromethanolic extract of M. vulgare had an anti-inflammatory effect, which corroborates the usefulness of this plant in the treatment of inflammatory disorders.

  14. Effect of Dark Chocolate Extracts on Phorbol 12-Myristate 13-Acetate-Induced Oxidative Burst in Leukocytes Isolated by Normo-Weight and Overweight/Obese Subjects

    Science.gov (United States)

    Ioannone, Francesca; Sacchetti, Giampiero; Serafini, Mauro

    2017-01-01

    Oxidative and inflammatory stress represents a major risk factor for cardiovascular disease (CVD) in overweight and obese subjects. Between the different plant foods, chocolate has been shown to decrease CVD risk due to its antioxidant and anti-inflammatory properties. However, as we recently showed in epidemiological studies, meta-analyses, and human trials, dietary antioxidants resulted more effective in subjects characterized by an ongoing oxidative stress, than in healthy people. Aim of this work was to investigate the effect of different concentrations of chocolate phenolic extract (CPE) on in vitro free radical production, stimulated by phorbol 12-myristate 13-acetate (PMA), in leukocytes extracted from blood of normo-weight and overweight/obese subjects. Neutrophils from overweight/obese group had a significantly higher free radical production compared to the normo-weight group. In neutrophils, the lowest CPE concentration significantly reduced free radical production in overweight/obese group only, and higher CPE concentrations were effective in both groups. In monocytes, the CPE concentration that was significantly effective in reducing free radical production was lower in overweight/obese subjects than in normo-weight subjects. Chocolate polyphenol extracts inhibit oxidative burst in human neutrophils and monocytes with a higher efficiency in subjects characterized by an unphysiological oxidative/inflammatory stress, such as overweight and obese. Results of this study provide further evidence about a differential role of dietary antioxidant strictly related to the “stress” condition of the subjects. PMID:28649567

  15. Effect of Dark Chocolate Extracts on Phorbol 12-Myristate 13-Acetate-Induced Oxidative Burst in Leukocytes Isolated by Normo-Weight and Overweight/Obese Subjects

    Directory of Open Access Journals (Sweden)

    Francesca Ioannone

    2017-06-01

    Full Text Available Oxidative and inflammatory stress represents a major risk factor for cardiovascular disease (CVD in overweight and obese subjects. Between the different plant foods, chocolate has been shown to decrease CVD risk due to its antioxidant and anti-inflammatory properties. However, as we recently showed in epidemiological studies, meta-analyses, and human trials, dietary antioxidants resulted more effective in subjects characterized by an ongoing oxidative stress, than in healthy people. Aim of this work was to investigate the effect of different concentrations of chocolate phenolic extract (CPE on in vitro free radical production, stimulated by phorbol 12-myristate 13-acetate (PMA, in leukocytes extracted from blood of normo-weight and overweight/obese subjects. Neutrophils from overweight/obese group had a significantly higher free radical production compared to the normo-weight group. In neutrophils, the lowest CPE concentration significantly reduced free radical production in overweight/obese group only, and higher CPE concentrations were effective in both groups. In monocytes, the CPE concentration that was significantly effective in reducing free radical production was lower in overweight/obese subjects than in normo-weight subjects. Chocolate polyphenol extracts inhibit oxidative burst in human neutrophils and monocytes with a higher efficiency in subjects characterized by an unphysiological oxidative/inflammatory stress, such as overweight and obese. Results of this study provide further evidence about a differential role of dietary antioxidant strictly related to the “stress” condition of the subjects.

  16. Technical characterization of dialysis fluid flow of newly developed dialyzers using mass transfer correlation equations.

    Science.gov (United States)

    Kunikata, Satoru; Fukuda, Makoto; Yamamoto, Ken-ichiro; Yagi, Yutaka; Matsuda, Masato; Sakai, Kiyotaka

    2009-01-01

    Dialysis fluid flow and mass transfer rate of newly developed dialyzers were evaluated using mass transfer correlation equations of dialysis fluid-side film coefficient. Aqueous creatinine clearance and overall mass transfer coefficient for APS-15S (Asahi Kasei Kuraray) as a conventional dialyzer, and APS-15SA (Asahi Kasei Kuraray), PES-150Salpha (Nipro), FPX140 (Fresenius), and CS-1.6U (Toray) as newly developed dialyzers were obtained at a blood-side flow rate (QB) of 200 ml/min, dialysis fluid-side flow rates (QD) of 200-800 ml/min and a net filtration rate (QF) of 0 ml/min. Mass transfer correlation equations between Sherwood number (Sh) containing dialysis fluid-side mass transfer film coefficient and Reynolds number (Re) were formed for each dialyzer. The exponents of Re were 0.62 for APS-15S whereas approximately 0.5 for the newly developed dialyzers. The dialysis fluid-side mass transfer film coefficients of the newly developed dialyzers were higher than those of the conventional dialyzer. Based on the mass transfer correlation equations, introduction of short taper, full baffle of dialyzer jacket and further wave-shaped hollow fiber improves the dialysis fluid flow of the newly developed dialyzers.

  17. Modified cellulosic dialyzer membranes: an investigative tool in thrombogenicity studies.

    Science.gov (United States)

    Mahiout, A; Meinhold, H; Kessel, M; Vienken, J; Baurmeister, U

    1988-01-01

    We have previously demonstrated that chemical modification of cellulosic membranes with dimethyl-amino-ethyl (DEAE) groups significantly improves membrane properties in terms of biocompatibility. Here, we show that DEAE substitution also alters the membrane's thrombogenic properties, and cellulosic membranes with various amounts of DEAE substitution were produced. Clinical dialyzers were constructed using two experimental membrane materials: modified cellulose-low (MC-low) and MC-high; standard unsubstituted cellulose was used as a control. Six patients were treated for a period of 3 weeks with each type of dialyzer and a heparin dose of less than 6000 IU/treatment. MC-low exhibited less extracorporeal beta-thromboglobulin and thromboxane B2 release than MC-high or Cuprophan. In addition, residual blood volume after clinical use was lower in the MC-low type. MC-low and MC-high induced less complement activation than Cuprophan, as characterized by extracorporeal C5a and C3a plasma concentrations (75% less C5a generation and 50 to 70% less C3a generation than unsubstituted cellulose).

  18. The Effect of Hydro-Alcoholic Extract of Foeniculum vulgare Mill on Leukocytes and Hematological Tests in Male Rats

    Science.gov (United States)

    Mansouri, Esrafil; Kooti, Wesam; Bazvand, Maryam; Ghasemi Boroon, Maryam; Amirzargar, Ashraf; Afrisham, Reza; Afzalzadeh, Mohammad Reza; Ashtary-Larky, Damoon; Jalali, Nasrin

    2015-01-01

    Background: Medicinal plants have a long history in treating blood disorders, which is one of the most common problems in today's advanced world. Fennel (Foeniculum vulgare) is a medicinal plant with a high content of polyphenols and has antioxidant and immunomodulatory properties. Objectives: The aim of this study was to evaluate the effect of hydro-alcoholic extract of fennel on some hematological indices in male rats. Materials and Methods: In this experimental study, thirty male Wistar rats were divided into six groups (five rats in each group). The first group (control) did not receive any dose; the second group (sham) received 1 mL normal saline (extraction solvent); and the experimental groups 1, 2, 3 and 4 respectively received 1 mL hydro alcoholic extract of fennel in four doses of 250, 500, 750 and 1000 mg/kg of body weight every 48 hours for 30 days by gavage. One day after the last gavage following induction of anesthesia and taking blood from the heart of rats, measurement of red and white blood cells, hemoglobin, hematocrit and tests of bleeding and coagulation time (CT) were performed. The data was analyzed by one-way ANOVA test using SPSS15 software. Results: Fennel increased mean RBC (7.54 ± 0.53 × 106) and WBC (5.89 ± 0.78 × 103) values, especially at a dose of 250 mg/mL and CT (2.45 ± 0.20) at a dose of 500mg/mL compared to the control group (P < 0.05). Conclusions: Fennel increased red and white blood cells probably due to the presence of polyphenols and antioxidant activity of fennel and reduced negative effects of free radicals on blood cells. PMID:25866717

  19. The cardio-renal-anaemia syndrome predicts survival in peritoneally dialyzed patients

    National Research Council Canada - National Science Library

    Malyszko, Jolanta; Zbroch, Edyta; Malyszko, Jacek; Mysliwiec, Michal; Iaina, Adrian

    2010-01-01

    .... The correction of anaemia was effective in the amelioration of both cardiac and renal failure. We studied the relationship between the severity of CRA syndrome in peritoneally dialyzed patients and their survival probability...

  20. Evaluation of dialyzer jacket structure and hollow-fiber dialysis membranes to achieve high dialysis performance.

    Science.gov (United States)

    Hirano, Ayaka; Yamamoto, Ken-ichiro; Matsuda, Masato; Ogawa, Takehito; Yakushiji, Taiji; Miyasaka, Takehiro; Sakai, Kiyotaka

    2011-02-01

    The objective of this study was to determine the optimum dialyzer jacket structure and hollow-fiber dialysis membrane, both of which are indispensable factors for achieving high dialysis performance, by clarifying the relationship between the dialysis performance and the flow of dialysate and blood in a hollow-fiber dialyzer. We evaluated the clearance, dialysate, and blood flow for four commercially available hollow-fiber dialyzers, namely, the APS-15S, APS-15SA, TS-1.6UL, and CX-1.6U. To evaluate dialysate and blood flow, we measured the residence-time distribution of dialysate and blood flow of these dialyzers by the pulse-response method. We also determined the clearances of urea, creatinine, vitamin B(12), and lysozyme to evaluate the dialysis performance of these dialyzers. While the baffle and taper structures allow effective supply of dialysate into the dialyzer jacket, the hollow-fiber shape, inner diameter, and packing density significantly influence the dialysate flow. In dialyzers with long taper-holding slits, the slit area is a key design parameter for achieving optimum dialysate flow. Similarly, the blood flow is significantly influenced by the structure of the inflowing and outflowing blood ports at the header of a dialyzer, and the shape and inner diameter of the hollow fibers. Hollow fibers with smaller inner diameters cause an increase in blood pressure, which causes blood to enter the hollow fibers more easily. The hollow-fiber shape hardly affects the blood flow. While improved dialysate and blood flow cause higher clearance of low molecular-weight substances, higher membrane area and pure-water permeability accelerate internal filtration, thereby causing an increase in the clearance of large molecular-weight substances. © 2010 The Authors. Therapeutic Apheresis and Dialysis © 2010 International Society for Apheresis.

  1. Diffusive clearance of small and middle-sized molecules in combined dialyzer flow configurations.

    Science.gov (United States)

    Eloot, S; De Vos, J Y; Hombrouckx, R; Verdonck, P

    2004-03-01

    Clearance of low (LMW) and middle molecular weight (MMW) solutes was investigated in vitro for different dialyzer configurations and mutual flow directions. Single pass tests were performed with two low flux Fresenius F6HPS hemodialyzers placed in series (12 tests) and in parallel (6 tests), and results were compared with those for one single dialyzer (2 tests). Either high concentrated (45mS/cm) bicarbonate dialysis fluid (surrogate LMW) or trisodiumphosphate (surrogate MMW) concentration (31mS/cm) was used as blood substitution fluid. Standard blood and dialysate flows of 250 and 500ml/min, respectively, were prescribed. Clearance was derived from conductivity measurements in blood and dialysate compartment, correcting for the overall ultrafiltration rate of 0.1-0.5l/h. In a single dialyzer, changing the counter current flow to co-current deteriorates diffusive clearance by 14% (LMW) and 18% (MMW). Compared to one single dialyzer using counter current flow, clearance increases by 3 to 8% (LMW) and by 15 to 18% (MMW) using two dialyzers in parallel and in series, respectively. As a consequence, the benefit by using a second dialyzer is more prominent for larger molecules. Moreover, pressure profiles drawn for the different configurations show the impact of limited convection on diffusive clearance.

  2. Preparation and Determination of Immunological Activities of Anti-HBV Egg Yolk Extraction

    Institute of Scientific and Technical Information of China (English)

    Yanping Xu; Weimin Zou; Xuejun Zhan; Shuhua Yang; Daze Xie; Sailiang Peng

    2006-01-01

    To prepare an effective immune preparation to treat hepatitis B, hens were immunized with hepatitis B vaccines,and then anti-HBV egg yolk extraction (anti-HBV EYE) was refined from egg yolk by a dialyzable method. Its chemical characteristics were identified by ultraviolet spectrum, HPLC, Lowry analysis and pharmacopocia-raleted methods. The specific immunological activity was examined by leukocyte adherence inhibition (LAI) in vitro and delayed type hypersensitivity (DTH) in vivo. Anti-HBV EYE was a small dialyzable substance with molecular weight less than 12 kD containing 18 kinds of amino acids. The preparation could obviously inhibit LAI and DTH which was similar to hepatitis B virus-specific transfer factor of pig spleen. However, there were no similar effects observed in the nonspecific transfer factor (NTF) group, control egg yolk extraction (CEYE) group and hepatitis A virus (HAV) group. The results suggested that anti-HBV EYE contained hepatitis B virus-specific transfer factor(STF) and had the antigen-specific cell immune activity similar to PSHBV-TF. The STF obtained from egg yolk of the hens immunized with specific antigen, might be a potential candidate for immunoregulation in hepatitis B prevention and treatment.

  3. Retention of cytokine-inducing substances inside high-flux dialyzers.

    Science.gov (United States)

    Lufft, V; Mahiout, A; Shaldon, S; Koch, K M; Schindler, R

    1996-01-01

    Reprocessing of dialyzers is often performed with nonsterile solutions possibly contaminated with bacterial-derived cytokine-inducing substances. We investigated the retention of cytokine-inducing substances inside the dialyzer during reprocessing in a closed loop in vitro hemodialysis system using a polyamide high flux membrane. After the first in vitro circulation of human whole blood, rinse of the blood compartment (BC) and reverse ultrafiltration (RUF) was performed with either cytokine-inducing substance-free saline or saline contaminated with filtrates from Pseudomonas cultures (6 ng/ml LAL-reactive material); subsequently, dialyzers were stored in 2% formaldehyde. Dialyzers were rinsed with approximately 15 liters pyrogen-free saline before the second circulation using blood from the same donor; the effluates were free of cytokine-inducing substances and formaldehyde. Before and after the blood circulations, peripheral blood mononuclear cells (PBMC) were separated and total production of IL-1 alpha and IL-1 beta was determined after overnight incubation. In noncirculated PBMC as well as in PBMC separated after whole blood circulation with pyrogen-free processed dialyzers, production of IL-1 beta was not detectable. After contaminated rinse of the BC, production of IL-1 beta could be observed (1,600 +/- 1,100 pg/ml, mean +/- SEM). When pyrogen-free RUF was performed after contaminated BC rinse, IL-1 beta production averaged 163 +/- 92 pg/ml when using reused dialyzers, but 1,820 +/- 880 pg/ml when using new dialyzers. After reuse with pyrogen-free BC-rinse and contaminated RUF no IL-1 beta synthesis was observed; however, when pyrogen-free BC-rinse and contaminated RUF was applied to new dialyzers, IL-1 beta synthesis averaged 1,620 +/- 1,200 pg/ml. We conclude that cytokine-inducing substances are retained inside the dialyzer, probably by adsorption to the membrane as well as to the protein layer covering the membrane and are still biologically active after

  4. Dual Stimulus-Dependent Effect of Oenothera paradoxa Extract on the Respiratory Burst in Human Leukocytes: Suppressing for Escherichia coli and Phorbol Myristate Acetate and Stimulating for Formyl-Methionyl-Leucyl-Phenylalanine

    Directory of Open Access Journals (Sweden)

    Izabela Burzynska-Pedziwiatr

    2014-01-01

    Full Text Available Although a growing body of evidence suggests that plant polyphenols can modulate human immune responses, their simultaneous action on monocyte and neutrophil oxidative burst is currently poorly understood. Based on the hypothesis that various polyphenols contained in plant extracts might affect the oxidative burst of phagocytes, we evaluated the effects of ethanolic O. paradoxa extract polyphenols on monocyte and neutrophil oxidative burst in vitro activated by different stimuli, including opsonized bacteria E. coli, phorbol 12-myristate 13-acetate (PMA, and formyl-methionyl-leucyl-phenylalanine (fMLP. Samples were analyzed by the dihydrorhodamine flow cytometry assay. Our results showed that the extract repressed significantly and dose-dependently reactive oxygen species production in both cell types stimulated with E. coli and PMA (P < 0.05 and its inhibitory efficiency was stimulus- and cell-type-dependent. Interestingly, there was significant stimulatory effect of the extract on bursting phagocytes induced by fMLP (P < 0.05. Additionally, several flavonoids and phenolic compounds as well as penta-galloyl-β-(D-glucose (PGG, the representative of hydrolyzable tannins, were identified in the 60% extract by high-performance liquid chromatography (HPLC coupled to electrospray ionization in negative ion mode. In summary, the ethanolic O. paradoxa extract, rich in flavonoids and phenolic compounds, exhibits dual stimulus-dependent effect on the respiratory burst in human leukocytes; hence, it might affect immune responses in humans.

  5. Leukocyte Activation and Circulating Leukocyte-Derived Microparticles in Preeclampsia

    NARCIS (Netherlands)

    Lok, Christianne A. R.; Jebbink, Jiska; Nieuwland, Rienk; Faas, Marijke M.; Boer, Kees; Sturk, Augueste; Van Der Post, Joris A. M.

    2009-01-01

    Preeclampsia shows characteristics of an inflammatory disease including leukocyte activation. Analyses of leukocyte-derived microparticles (MP) and mRNA expression of inflammation-related genes in leukocytes may establish which subgroups of leukocytes contribute to the development of preeclampsia. B

  6. Milk peptides increase iron dialyzability in water but do not affect DMT-1 expression in Caco-2 cells.

    Science.gov (United States)

    Argyri, Konstantina; Tako, Elad; Miller, Dennis D; Glahn, Raymond P; Komaitis, Michael; Kapsokefalou, Maria

    2009-02-25

    In vitro digestion of milk produces peptide fractions that enhance iron uptake by Caco-2 cells. The objectives of this study were to investigate whether these fractions (a) exert their effect by increasing relative gene expression of DMT-1 in Caco-2 cells and (b) enhance iron dialyzability when added in meals. Two milk peptide fractions that solubilize iron were isolated by Sephadex G-25 gel filtration of a milk digest. These peptide fractions did not affect relative gene expression of DMT-1 when incubated with Caco-2 cells for 2 or 48 h. Dialyzability was measured after in vitro simulated gastric and pancreatic digestion. Both peptide fractions enhanced the dialyzability of iron from ferric chloride added to PIPES buffer, but had no effect on dialyzability from milk or a vegetable or fruit meal after in vitro simulated gastric and pancreatic digestion. However, dialyzability from milk was enhanced by the addition of a more concentrated lyophilized peptide fraction.

  7. Leukocytes in capillary flow.

    Science.gov (United States)

    Schmid-Schönbein, G W; Lee, J

    1995-01-01

    During disease, the flow of blood cells through the capillary network is one of the most perilous events in the microcirculation. Capillary distensibility, cytoplasmic activity of endothelial cells, red cells and leukocytes play an important role in capillary perfusion. Occlusion of capillaries is one of the early signs of vascular failure and is encountered in many different conditions and organs. Adhesion of leukocytes to the endothelium via expression of membrane adhesion molecules leads to microvascular entrapment with capillary occlusion.

  8. Membrane fouling and dialysate flow pattern in an internal filtration-enhancing dialyzer.

    Science.gov (United States)

    Yamamoto, Ken-ichiro; Hiwatari, Michihito; Kohori, Fukashi; Sakai, Kiyotaka; Fukuda, Makoto; Hiyoshi, Tatsuo

    2005-01-01

    For efficient removal of large molecular weight solutes by dialysis, several types of internal filtration-enhancing dialyzers (IFEDs) are commercially available. However, in a pressure-driven membrane separation process (i.e., filtration), membrane fouling caused by adhesion of plasma proteins is a severe problem. The objective of the present study is to investigate the effects of internal filtration on membrane fouling based on the membrane's pure-water permeability, diffusive permeability, and sieving coefficient. Hemodialysis experiments were performed with two different dialyzers, IFEDs and non-IFEDs. Local membrane fouling in each dialyzer was evaluated by measuring the pure-water permeability, the diffusive permeability, and the sieving coefficient of native membranes and membranes treated with bovine blood. The effects of packing ratio on dialysate flow pattern were also evaluated by measuring the time required for an ion tracer to reach electrodes placed in the dialyzers. In the IFED, membrane fouling caused by protein adhesion is increased because of enhanced internal filtration only at the early stage of dialysis, and this fouling tends to occur only near the dialysate outlet port. However, enhanced internal filtration has little effect on measured membrane transfer parameters.

  9. Intravital leukocyte detection using the gradient inverse coefficient of variation.

    Science.gov (United States)

    Dong, Gang; Ray, Nilanjan; Acton, Scott T

    2005-07-01

    The problem of identifying and counting rolling leukocytes within intravital microscopy is of both theoretical and practical interest. Currently, methods exist for tracking rolling leukocytes in vivo, but these methods rely on manual detection of the cells. In this paper we propose a technique for accurately detecting rolling leukocytes based on Bayesian classification. The classification depends on a feature score, the gradient inverse coefficient of variation (GICOV), which serves to discriminate rolling leukocytes from a cluttered environment. The leukocyte detection process consists of three sequential steps: the first step utilizes an ellipse matching algorithm to coarsely identify the leukocytes by finding the ellipses with a locally maximal GICOV. In the second step, starting from each of the ellipses found in the first step, a B-spline snake is evolved to refine the leukocytes boundaries by maximizing the associated GICOV score. The third and final step retains only the extracted contours that have a GICOV score above the analytically determined threshold. Experimental results using 327 rolling leukocytes were compared to those of human experts and currently used methods. The proposed GICOV method achieves 78.6% leukocyte detection accuracy with 13.1% false alarm rate.

  10. Influência de extratos hidroetanólicos de plantas medicinais sobre a quimiotaxia de leucócitos humanos Influence of some medicinal plant hydroethanolic extracts on human leukocyte chemotaxis

    Directory of Open Access Journals (Sweden)

    M. M. Presibella

    2003-12-01

    leukocytes migration induced by casein, using the Boyden chamber method. Dexamethasone has been used as a positive control for leukocyte inhibition in the same experimental approach. The data herein presented showed a significant inhibition of the casein-induced polymorphonuclear leukocytes migration for all plants studied. However, the intensity of such activity was variable according to the dose and plant tested. For pau-pra-tudo, canela-de-veado, and cipó-suma extracts the average number of migrated polymorphonuclear leukocytes was 81.6±3.9%, 85.4±2.4% and 91.7±2.2% of the input for the doses of 1000, 10, and 1mg/ml, respectively, while for dexamethasone, the value found was 70.3±5.9%. Although further studies are needed, the results presented in this study may be useful to clarify the anti-inflammatory properties of these herbal medicines, supporting their ethnobothanical use for the treatment of inflammatory diseases.

  11. Internal filtration, filtration fraction, and blood flow resistance in high- and low-flux dialyzers.

    Science.gov (United States)

    Schneditz, Daniel; Zierler, Edda; Vanholder, Raymond; Eloot, Sunny

    2014-01-01

    It was the aim to examine the fluid flow in blood and dialysate compartments of highly permeable hollow fiber dialyzers where internal filtration contributes to solute removal but where excessive filtration bears a risk of cell activation and damage. Flow characteristics of high- (HF) and low-flux (LF) dialyzers were studied in lab-bench experiments using whole bovine blood. Measurements obtained under different operating conditions and under zero net ultrafiltration were compared to theoretical calculations obtained from a mathematical model. Experimental resistances in the blood compartment were within ±2% of those calculated from the model when dialysate was used as a test fluid. With whole blood, the experimental resistances in the blood compartment were only 81.8 ± 2.8% and 83.7 ± 4.3% of those calculated for the LF and HF dialyzer, respectively. Surprisingly, measured blood flow resistance slightly but significantly decreased with increasing flow rate (p filtration fraction, while overall internal filtration increased. The increase in internal filtration when increasing blood flow is associated with a beneficial reduction in internal filtration fraction. Concerns of increased hemoconcentration when increasing blood flow therefore appear to be unwarranted.

  12. Serum somatomedin A and non-dialyzable urinary hydroxyproline in girls with idiopathic scoliosis.

    Science.gov (United States)

    Skogland, L B; Miller, J A; Skottner, A; Fryklund, L

    1981-06-01

    Serum somatomedin A was determined by radioreceptor assay in 31 girls with idiopathic scoliosis and 30 controls, with ages ranging from 9.7-16.2 years. In the material as a whole no significant difference could be found between scoliotics and controls. Similarly no differences were found in the girls under 13 years of age. However, in girls older than 13 years the controls had significantly higher somatomedin A levels than the scoliotics. In the second part of the study 26 girls with idiopathic scoliosis and 22 controls were examined with regard to the urinary excretion of non-dialyzable hydroxyproline,. which has been suggested to be correlated with collagen synthesis. The percentage non-dialyzable fraction of the total hydroxyproline was found to be significantly higher in the control girls. The mean age was 12.8 years for both groups. The results suggest a higher growth rate in the controls than in the scoliotics for girls over 12 years. There is, however, no definite evidence as to the function of the somatomedins or of the relationship between non-dialyzable hydroxyproline and growth.

  13. Extraction of Leukocyte Areas Based on Improved ITTI and Particle Swarm Optimization Algorithm%基于改进ITTI模型及粒子群优化算法的白细胞区域提取

    Institute of Scientific and Technical Information of China (English)

    纪滨; 杨盼盼; 申元霞

    2016-01-01

    白细胞显微图像病理分析中,人眼关注的白细胞是感兴趣的区域。ITTI视觉模型是提取图像感兴趣区域(ROI)的有效办法。为了进一步改善其提取的准确性,提出了基于改进的ITTI视觉模型与粒子群优化算法相结合的目标控制方法,并将其应用于医学骨髓细胞图像中的白细胞区域提取。首先利用高斯滤波和多尺度归一化的方法分别提取原始图像的方向、亮度、颜色显著性特征,再根据人眼的视觉对不同显著性特征敏感程度不同的特性对3种显著性特征采用自适应系数相融合的方式得到显著图,最后利用基于改进的粒子群优化算法的Otsu法对显著图进行ROI的提取,并采用数字形态学的方法对其进行后续处理。结果表明,本文算法可以较好地提取完整的白细胞区域,有助于提高病理分析的效率。%In the process of pathological analysis of the Leukocyte microscopic image, the leukocyte areas are regions of interest(ROI). The ITTI visual model is an effective method of extracting the ROI from image. For improving the extracting accuracy, an object extracting method combining the improved ITTI visual model with particle swarm optimization algorithm is proposed and used to extract the ROI from the bone marrow cell image. Firstly, based on Gaussian filter and multi-scale normalization, features of the orientation, brightness, and color are computed from the original image. And then, according the fact that the sensitivity of eyes is not the same as different features the saliency map is obtained with adaptive coefficient from three significant characteristics. Finally, by using Otsu method based on the improved particle swarm optimization(PSO) algorithm, the ROIs are extracted and subsequent processed with method of morphology. Experimental results show that this method can extract white blood cell areas perfectly, which is helpful for improving the efficiency of

  14. The Effect of Vitamin E-Modified Dialyzers on Acute Intra-dialytic Symptoms: A Comparative Crossover Study

    Directory of Open Access Journals (Sweden)

    Tanimu Danlami

    2000-01-01

    Full Text Available We performed a crossover study to compare the effect of vitamin E-modified dialyzers on acute intra-dialytic symptoms, with other membranes. Twenty patients on hemodialysis were studied. They were divided into two equal groups of low-flux (C15NL, E15NL and high-flux (F60, EE15NL membrane dialyzers. Within each group, a vitamin E-modified dialyzer was compared with another dialyzer in a crossover design over a two-month period. All study patients were seen during each dialysis session by a physician and the occurrence of intra-dialytic symptoms were recorded. There was a significant overall improvement in the incidence of acute intra-dialytic symptoms with the use of vitamin E-modified dialyzers as compared with the other membranes. This effect was more for cuprophane than polysulfone. The occurrence of hypotensive episodes did not differ. Our study indicates that we can achieve a reduction in the incidence of acute intra-dialytic symptoms with the use of vitamin E-modified membrane as compared to cuprophane and polysulfone.

  15. Characterization of insulin adsorption behavior of dialyzer membranes used in hemodialysis.

    Science.gov (United States)

    Abe, Masanori; Okada, Kazuyoshi; Ikeda, Kazuya; Matsumoto, Shiro; Soma, Masayoshi; Matsumoto, Koichi

    2011-04-01

    Although it has been reported that plasma insulin is removed by hemodialysis (HD), the mechanism for this has not been elucidated. We investigated the mechanism of insulin removal during HD treatment and the characteristics of insulin removal with three high-flux membranes. In our in vivo study, 20 stable diabetic patients on HD were randomly selected for three HD sessions with three different membranes: polysulfone (PS), cellulose triacetate (CTA), and polyester polymer alloy (PEPA). Blood samples were obtained from the blood tubing at the arterial (A) site at the beginning and end of the sixth HD session to investigate insulin reduction in patients. At 1 h after the initiation of dialysis, blood samples were obtained from both the A and venous sites of the dialyzer to investigate the insulin clearance with the different membranes. There was a significant reduction in patients' plasma insulin at each time point with each of the three membranes. The insulin clearance with the PS membrane was significantly higher than that with the CTA and PEPA membranes. Although no difference was observed in the plasma insulin reduction rate between the three membranes in the total subject group, there was a significantly higher reduction rate with the PS membrane in insulin-dependent diabetes mellitus subjects. The clearance of insulin in in vitro tests was significantly higher with the PS and PEPA membranes than with the CTA membrane in both new and clinically used dialyzers. Insulin was not detected in the dialysate or ultrafiltration fluids in either the in vivo or in vitro studies. The mechanism of plasma insulin clearance by HD is mainly by adsorption, and the amount of insulin adsorbed differed depending on the dialyzer membrane used. © 2011, Copyright the Authors. Artificial Organs © 2011, International Center for Artificial Organs and Transplantation and Wiley Periodicals, Inc.

  16. Dialyzer membranes: effect of surface area and chemical modification of cellulose on complement and platelet activation.

    Science.gov (United States)

    Mahiout, A; Meinhold, H; Kessel, M; Schulze, H; Baurmeister, U

    1987-04-01

    Using an ex vivo model, the effects of membrane composition and surface area on both the complement system (as reflected by plasma C3a levels) and platelets [as indicated by plasma concentrations of thromboxane B2 (TXB2) and platelet factor 4 (PF4)] were studied. In this model, polyacrylonitrile (PAN) was associated with less complement activation than cuprammonium cellulose (CC). A new "modified cellulose" (MC) membrane, in which a small number of the free hydroxyl groups on cellulose are substituted with a tertiary amino compound, was also associated with a low degree of complement activation, similar to that with PAN. However, the extent of hydroxyl group substitution in four MC membrane subtypes did not correlate with the reduction in complement activation. In studies using CC, the amount of generated C3a correlated with the membrane surface area, although the relationship was curvilinear. Plasma concentrations at the "dialyzer" outlet of TXB2 and PF4 were similar with CC, PAN, and MC. In studies with the MC subtypes, increasing the extent of hydroxyl group substitution paradoxically increased, albeit slightly, the amount of TXB2 generation. In studies with CC, a linear relationship between membrane surface area and TXB2 generation was found. The results suggest a dissociation between platelet and complement effects among different dialyzer membranes, and underline the importance of membrane surface area.

  17. Nickel levels in convenience and fast foods: In vitro study of the dialyzable fraction

    Energy Technology Data Exchange (ETDEWEB)

    Cabrera-Vique, Carmen, E-mail: carmenc@ugr.es [Department of Nutrition and Bromatology, University of Granada, Granada (Spain); Mesias, Marta [Department of Nutrition and Bromatology, University of Granada, Granada (Spain); Bouzas, Paula R. [Department of Statistic, University of Granada, Granada (Spain)

    2011-03-15

    Nickel presence was determined in 170 samples of 43 different convenience and fast foods widely consumed in Spain. Electrothermal atomic absorption spectrometry was used as analytical technique. Reliability of the procedure was checked. Ni levels ranged from 18.50 to 95.00 ng g{sup -1} (fresh weight of edible portion). The most elevated Ni concentrations were found in egg- and pork-based foods and in sauces but there is a high variability inside of each one of these foods. Ni content increases in products that contain spices and aromatic herbs, whole cereals, dry fruits, cheese and mushrooms. Mean Ni dialyzable fraction estimated by in vitro assays ranged from 4.50 to 7.75%. This study shows that the probability of exposure to health risks from these foods is overall small. However, the present findings are of potential use in food composition tables and to estimate the Ni dietary intake and tolerable intake levels in accordance with the current dietary habits. - Research highlights: {yields}Ni levels in convenience and fast food range from 18.50 to 95.00 ng/g. {yields}Ni content increases in products that contain spices and aromatic herbs, whole cereals, dry fruits, cheese and mushrooms. {yields}Mean Ni dialyzable fraction estimated by in vitro assays ranges from 4.50 to 7.75%. {yields}The probability of exposure to health risks from convenience and fast food is overall small.

  18. Use of ascorbic and citric acids to increase dialyzable iron from vinal (Prosopis ruscifolia) pulp.

    Science.gov (United States)

    Bernardi, C; Freyre, M; Sambucetti, M E; Pirovani, M E

    2004-01-01

    Vinal (Prosopis ruscifolia) is an ecologically important wild leguminous tree that grows spontaneously in Argentine deforested lands, the fruit of which is consumed by humans and animals. Because considerable iron content with low to intermediate availability has been previously reported in vinal pulps, its enhancement would be of interest. Iron availability was determined as iron dialyzability using an in vitro technique. Response surface methodology was used to evaluate iron availability increase after adding ascorbic and/or citric acids to vinal pulp at different mM acid/mM Fe ratios. Those ratios ranged from 0.05:1 to 9.95:1 and from 0.5:1 to 99.5:1 for ascorbic acid/Fe (AA:Fe) and citric acid/Fe (CA:Fe), respectively. The obtained second- and first-order polynomial equations showed that AA:Fe and CA:Fe molar ratios linear terms had a significant effect on iron dialyzability increase (P iron availability to a maximum of 4.6 times. Additional confirmatory experiments were made adding the same organic acids to different vinal pulps and to a traditional cake prepared with vinal pulp called "patay." There were no significant differences (p > 0.05) between predicted values obtained by the model and experimental results.

  19. Metallothionein mediates leukocyte chemotaxis

    Directory of Open Access Journals (Sweden)

    Lynes Michael A

    2005-09-01

    Full Text Available Abstract Background Metallothionein (MT is a cysteine-rich, metal-binding protein that can be induced by a variety of agents. Modulation of MT levels has also been shown to alter specific immune functions. We have noticed that the MT genes map close to the chemokines Ccl17 and Cx3cl1. Cysteine motifs that characterize these chemokines are also found in the MT sequence suggesting that MT might also act as a chemotactic factor. Results In the experiments reported here, we show that immune cells migrate chemotactically in the presence of a gradient of MT. This response can be specifically blocked by two different monoclonal anti-MT antibodies. Exposure of cells to MT also leads to a rapid increase in F-actin content. Incubation of Jurkat T cells with cholera toxin or pertussis toxin completely abrogates the chemotactic response to MT. Thus MT may act via G-protein coupled receptors and through the cyclic AMP signaling pathway to initiate chemotaxis. Conclusion These results suggest that, under inflammatory conditions, metallothionein in the extracellular environment may support the beneficial movement of leukocytes to the site of inflammation. MT may therefore represent a "danger signal"; modifying the character of the immune response when cells sense cellular stress. Elevated metallothionein produced in the context of exposure to environmental toxicants, or as a result of chronic inflammatory disease, may alter the normal chemotactic responses that regulate leukocyte trafficking. Thus, MT synthesis may represent an important factor in immunomodulation that is associated with autoimmune disease and toxicant exposure.

  20. Pioglitazone improves insulin sensitivity, reduces visceral fat and stimulates lipolysis in non diabetic dialyzed patients

    Directory of Open Access Journals (Sweden)

    Anne Zanchi

    2012-06-01

    Full Text Available Insulin resistance is common in dialyzed patients and is associated with increased mortality and protein-energy wasting. The aim of this study was to investigate the effect of pioglitazone (PIO, a powerful insulin sensitizer, on insulin sensitivity, body composition and adipose tissue metabolism, in dialyzed patients. A double blind randomized cross-over study was performed in non diabetic dialysis patients. Each patient followed 2 treatment phases of 16 weeks, starting either with oral PIO 45 mg/d or placebo (PL, and then switched to the other phase. At the end of each phase, patients underwent hyperinsulinemic euglycemic clamps, dual energy X-ray absorptiometry, an abdominal CT, and extensive plasma biochemical analysis. Twelve patients including 8 HD (59.6±4.4 y and 4 PD patients (43.5±3.6 y were recruited. Nine patients completed both phases and 3 patients dropped out (renal transplantation/2 HD and peritonitis/1 PD. PIO was safe and well tolerated. Under PIO, insulin sensitivity improved, as assessed by increased total glucose disposal rate (1.98±0.24 for PIO versus 1.58±0.12 umol/kg/min for PL, p<0.05, and reduced glucose endogenous hepatic production. PIO did not affect post-dialysis body weight, total fat and lean body mass, but significantly reduced visceral adipose tissue (VAT area and the VAT/SAT (subcutaneous adipose tissue ratio. HDL-cholesterol significantly increased. PIO decreased CRP (3.96±1.44 mg/l vs 7.88±2.56, p<0.05, plasma leptin, and dramatically reduced leptin/adiponectin ratio. Glycerol turnover, circulating glycerol and non esterified fatty acids were paradoxically increased. In conclusion, the improvement in insulin sensitivity by PIO, in non diabetic dialyzed patients, was associated with favorable metabolic effects, reduction in inflammation and body fat redistribution. The stimulation of systemic lipolysis was a surprising finding which may reflect adipose tissue remodeling and/or a paradoxical lypolitic

  1. Differences in perceived health status between kidney transplant recipients and dialyzed patients are based mainly on the selection process

    NARCIS (Netherlands)

    Rosenberger, Jaroslav; van Dijk, Jitse P.; Prihodova, Lucia; Majernikova, Maria; Nagyova, Iveta; Geckova, Andrea Madarasova; Roland, Robert; van den Heuvel, Wim J. A.; Groothoff, Johan W.

    2010-01-01

    Kidney transplantation offers longer survival, less morbidity and lower costs than dialysis. It is also believed to improve quality of life. The aim of this study was to compare prospectively the perceived health status (PHS) of dialyzed patients on a waiting list with kidney transplant recipients a

  2. Effect of blood flow rate on internal filtration in a high-flux dialyzer with polysulfone membrane.

    Science.gov (United States)

    Sakiyama, Ryoichi; Ishimori, Isamu; Akiba, Takashi; Mineshima, Michio

    2012-09-01

    Internal filtration/backfiltration (IF/BF) of a dialyzer depends on several parameters. This study evaluated the effect of the blood flow rate (Q (B)) on the internal filtration flow rate (Q (IF)) measured using Doppler ultrasonography for a high-flux dialyzer with a polysulfone membrane, APS-15E. In an in vitro study, bovine blood was circulated through the dialyzer, at a Q (B) of 100-350 mL/min. The clearances (CL) of creatinine, β(2)-microglobulin, and α(1)-microglobulin were then investigated. Q (IF) increased with the Q (B) value. A good correlation was obtained between Q (IF) and the pressure difference between the pressures at the inlet of the blood compartment and the pressure at the outlet of the dialysate compartment. The creatinine CL values strongly depended on Q (B) because molecular diffusion was dominant. The β(2)-microglobulin CL also depended on Q (B), because its removal rate seemed to be affected by both diffusive and convective transport caused by the IF/BF. An extremely low CL value was obtained for α(1)-microglobulin because of its low diffusivity and membrane fouling induced by proteins plugging the membrane. In conclusion, the IF/BF in the dialyzer strongly depends on Q (B). Furthermore, the dependence of the solute clearance on Q (B) decreased with increasing molecular size of the solute because of the decrease in diffusivity through the membrane.

  3. Development of bactericidal capacity and phagocytosis-associated metabolism of fetal pig leukocytes.

    Science.gov (United States)

    Holmes, B; Day, N; Haseman, J; Good, R A

    1972-02-01

    Evidence that the bactericidal ability and the stimulated oxidative metabolism of leukocytes appear in parallel during fetal development of the Minnesota Miniature pig has been obtained by application of the techniques applied to studies of human cells. It was demonstrated that leukocytes from 87- to 90-day fetuses were fully capable of ingesting Staphylococcus aureus but greatly diminished in bactericidal capacity as compared to leukocytes of older fetuses and adults. Although resting levels of oxygen consumption and hexose monophosphate pathway activity of leukocytes from the younger fetuses compared well with those of leukocytes from older animals, the phagocytosis-stimulated increments of metabolism were much less at 87 to 90 days of gestation than at later developmental stages. Both bactericidal capacity and increased metabolism of leukocytes reach adult levels by 100 days of gestation (normal gestation period of 115 to 120 days). Acrylamide gels stained for reduced nicotinamide adenine dinucleotide (NADH) and NADH phosphate (NADPH) diaphorase activity after disc electrophoresis of leukocyte extracts revealed normal mobility and intensity of NADH diaphorase bands. Three NADPH diaphorase bands were present in adult leukocyte extracts. Only the fast-migrating NADPH diaphorase band of 87- to 90-day cells stained with decreased intensity. This "deficiency" was no longer present at the later fetal period. The fast-migrating NADPH diaphorase band may represent an electron transfer protein which functions in cyanide-insensitive respiration of the leukocytes of the pig.

  4. Integrin Regulation during Leukocyte Recruitment.

    Science.gov (United States)

    Herter, Jan; Zarbock, Alexander

    2013-05-01

    Integrins are recognized as vital players in leukocyte recruitment. Integrin malfunction causes severe disease patterns characterized by the inability to fight pathogens. Although inflammatory reactions are beneficial and necessary for host defense, these reactions have to be controlled to prevent tissue destruction and harmful sequelae. In this review, we discuss the different signaling pathways leading to the change of integrin adhesiveness in neutrophils, monocytes, and lymphocytes. We thereby focus on the importance of integrin activation for the different steps of the leukocyte recruitment cascade, including rolling, adhesion, postadhesion strengthening, intravascular crawling, and transmigration, as each step necessitates the proper functioning of a distinct set of integrin molecules that has to be activated specifically. Additionally, we discuss endogenous mechanisms that balance and counteract integrin activation and limit leukocyte recruitment at the site of inflammation. Further insight into these complex mechanisms may provide new approaches for developing new anti-inflammatory therapies.

  5. An experimental and numerical study of the flow and mass transfer in a model of the wearable artificial kidney dialyzer

    Directory of Open Access Journals (Sweden)

    Rosenfeld Moshe

    2010-05-01

    Full Text Available Abstract Background Published studies of the past decades have established that mass transfer across the dialyzer membrane is governed by diffusion, convection and osmosis. While the former is independent of the pressure in the liquids, the latter two are pressure dependent and are enhanced when the pressure difference across the membrane is increased. The goal of the present study is to examine the impact of pulsatile flow on the transport phenomena across the membrane of a high-flux dialyzer in a wearable artificial kidney (WAK with a novel single small battery-operated pulsatile pump that drives both the blood and dialysate in a counter-phased manner, maximizing the trans-membrane pressure. Methods Both in-vitro experimental and numerical tools are employed to compare the performance of the pulsatile WAK dialyzer with a traditional design of a single-channel roller blood pump together with a centrifugal pump that drives the dialysate flow. The numerical methods utilize the axisymmetric Navier-Stokes and mass transfer equations to model the flow in the fibers of the dialyzer. Results While diffusion is still the dominating transport regime, the WAK pump enhances substantially the trans-membrane pressure and thus increases mass convection that might be as high as 30% of the overall transfer. This increase is obtained due to the design of the pulsatile WAK pump that increases ultrafiltration by increasing the trans-membrane pressure. Conclusions The experimental and numerical results revealed that when pumping at similar flow rates, a small battery-operated pulsatile pump provides clearances of urea and creatinine similar as or better than a large heavy AC-powered roller pump.

  6. Vitamin K status in peritoneally dialyzed patients with chronic kidney disease.

    Science.gov (United States)

    Stankowiak-Kulpa, Hanna; Krzyżanowska, Patrycja; Kozioł, Lidia; Grzymisławski, Marian; Wanic-Kossowska, Maria; Moczko, Jerzy; Walkowiak, Jarosław

    2011-01-01

    Abnormal vitamin K status was documented in patients with chronic kidney diseases (CKD), especially those undergoing hemodialysis. The data related to patients undergoing peritoneal dialysis (PD) are contradictory. Therefore, in the present study we aimed to evaluate vitamin K status in patients with CKD who are treated with continuous ambulatory PD. Twenty-eight patients entered into the study. Dialysis vintage ranged from 3 to 89 months. Vitamin K status was assessed in all subjects using undercarboxylated prothrombin measurement (PIVKA-II). In addition, total protein and albumin levels, total cholesterol, LDL cholesterol, triglyceride, calcium, urea and creatinine concentrations were determined. PIVKA-II concentrations were abnormal in 13 (46.4 %) subjects. BMI values, both total and LDL cholesterol concentrations were significantly higher in patients with than those without vitamin K deficiency. Moreover, PIVKA II levels correlated with BMI values (r = 0.441, p < 0.019), LDL cholesterol (r = 0.434, p < 0.021) and creatinine (r = 0.406, p< 0.032) concentrations. However, through the use of logistic regression analysis and multiple regression analysis, no clinical factor was documented to be the independent risk factor of vitamin K deficiency. In conclusion, vitamin K deficiency is a frequent condition in peritoneally dialyzed patients. Assessment of vitamin K status should become a standard procedure in this group of patients.

  7. Interstitial leukocyte migration and immune function.

    NARCIS (Netherlands)

    Friedl, P.H.A.; Weigelin, B.

    2008-01-01

    The trafficking of leukocytes into and within lymphoid and peripheral tissues is central to immune cell development, immunosurveillance and effector function. Interstitial leukocyte trafficking is the result of amoeboid polarization and migration, guided by soluble or tissue-bound chemoattractant

  8. Leukocyte adhesion - a fundamental process in leukocyte physiology

    Directory of Open Access Journals (Sweden)

    Gahmberg C.G.

    1999-01-01

    Full Text Available Leukocyte adhesion is of pivotal functional importance. The adhesion involves several different adhesion molecules, the most important of which are the leukocyte ß2-integrins (CD11/CD18, the intercellular adhesion molecules, and the selectins. We and others have extensively studied the specificity and binding sites in the integrins and the intercellular adhesion molecules for their receptors and ligands. The integrins have to become activated to exert their functions but the possible mechanisms of activation remain poorly understood. Importantly, a few novel intercellular adhesion molecules have been recently described, which seem to function only in specific tissues. Furthermore, it is becoming increasingly apparent that changes in integrins and intercellular adhesion molecules are associated with a number of acute and chronic diseases.

  9. Hemocompatible polyethersulfone/polyurethane composite membrane for high-performance antifouling and antithrombotic dialyzer.

    Science.gov (United States)

    Yin, Zehua; Cheng, Chong; Qin, Hui; Nie, Chuanxiong; He, Chao; Zhao, Changsheng

    2015-01-01

    Researches on blood purification membranes are fuelled by diverse clinical needs, such as hemodialysis, hemodiafiltration, hemofiltration, plasmapheresis, and plasma collection. To approach high-performance dialyzer, the integrated antifouling and antithrombotic properties are highly necessary for the design/modification of advanced artificial membranes. In this study, we propose and demonstrate that the physical blend of triblock polyurethane (PU) and polyethersulfone (PES) may advance the performance of hemodialysis membranes with greatly enhanced blood compatibility. It was found that the triblock PU could be blended with PES at high ratio owing to their excellent miscibility. The surfaces of the PES/PU composite membranes were characterized using attenuated total reflectance-Fourier transform infrared spectroscopy, X-ray photoelectron spectroscopy, water contact angle measurement, and surface ζ-potentials. The results indicated that the membrane surfaces were assembled with hydrophilic segregation layer owing to the migration of amphiphilic PU segments during membrane preparation, which might confer the composite membranes with superior hemocompatibility. The cross-section scanning electron microscopy images of the composite membranes exhibited structure transformation from finger-like structure to sponge-like structure, which indicated that the composite membrane had tunable porosity and permeability. The further ultrafiltration experiments indicated that the composite membranes showed increased permeability and excellent antifouling ability. The blood compatibility observation indicated that PES/PU composite membranes owned decreased protein adsorption, suppressed platelet adhesion, and prolonged plasma recalcification time. These results indicated that the PES/PU composite membranes exhibited enhanced antifouling and antithrombotic properties than the pristine PES membrane. The strategy may forward the fabrication of blood compatible composite membranes for

  10. EXTRACT

    DEFF Research Database (Denmark)

    Pafilis, Evangelos; Buttigieg, Pier Luigi; Ferrell, Barbra

    2016-01-01

    The microbial and molecular ecology research communities have made substantial progress on developing standards for annotating samples with environment metadata. However, sample manual annotation is a highly labor intensive process and requires familiarity with the terminologies used. We have the...... and text-mining-assisted curation revealed that EXTRACT speeds up annotation by 15-25% and helps curators to detect terms that would otherwise have been missed.Database URL: https://extract.hcmr.gr/....

  11. Adjusting MtDNA Quantification in Whole Blood for Peripheral Blood Platelet and Leukocyte Counts.

    Science.gov (United States)

    Hurtado-Roca, Yamilee; Ledesma, Marta; Gonzalez-Lazaro, Monica; Moreno-Loshuertos, Raquel; Fernandez-Silva, Patricio; Enriquez, Jose Antonio; Laclaustra, Martin

    2016-01-01

    Alterations of mitochondrial DNA copy number (mtDNAcn) in the blood (mitochondrial to nuclear DNA ratio) appear associated with several systemic diseases, including primary mitochondrial disorders, carcinogenesis, and hematologic diseases. Measuring mtDNAcn in DNA extracted from whole blood (WB) instead of from peripheral blood mononuclear cells or buffy coat may yield different results due to mitochondrial DNA present in platelets. The aim of this work is to quantify the contribution of platelets to mtDNAcn in whole blood [mtDNAcn(WB)] and to propose a correction formula to estimate leukocytes' mtDNAcn [mtDNAcn(L)] from mtDNAcn(WB). Blood samples from 10 healthy adults were combined with platelet-enriched plasma and saline solution to produce artificial blood preparations. Aliquots of each sample were combined with five different platelet concentrations. In 46 of these blood preparations, mtDNAcn was measured by qPCR. MtDNAcn(WB) increased 1.07 (95%CI 0.86, 1.29; p<0.001) per 1000 platelets present in the preparation. We proved that leukocyte count should also be taken into account as mtDNAcn(WB) was inversely associated with leukocyte count; it increased 1.10 (95%CI 0.95, 1.25, p<0.001) per unit increase of the ratio between platelet and leukocyte counts. If hematological measurements are available, subtracting 1.10 the platelets/leukocyte ratio from mtDNAcn(WB) may serve as an estimation for mtDNAcn(L). Both platelet and leukocyte counts in the sample are important sources of variation if comparing mtDNAcn among groups of patients when mtDNAcn is measured in DNA extracted from whole blood. Not taking the platelet/leukocyte ratio into account in whole blood measurements, may lead to overestimation and misclassification if interpreted as leukocytes' mtDNAcn.

  12. Adjusting MtDNA Quantification in Whole Blood for Peripheral Blood Platelet and Leukocyte Counts

    Science.gov (United States)

    Gonzalez-Lazaro, Monica; Moreno-Loshuertos, Raquel; Fernandez-Silva, Patricio; Enriquez, Jose Antonio; Laclaustra, Martin

    2016-01-01

    Alterations of mitochondrial DNA copy number (mtDNAcn) in the blood (mitochondrial to nuclear DNA ratio) appear associated with several systemic diseases, including primary mitochondrial disorders, carcinogenesis, and hematologic diseases. Measuring mtDNAcn in DNA extracted from whole blood (WB) instead of from peripheral blood mononuclear cells or buffy coat may yield different results due to mitochondrial DNA present in platelets. The aim of this work is to quantify the contribution of platelets to mtDNAcn in whole blood [mtDNAcn(WB)] and to propose a correction formula to estimate leukocytes' mtDNAcn [mtDNAcn(L)] from mtDNAcn(WB). Blood samples from 10 healthy adults were combined with platelet-enriched plasma and saline solution to produce artificial blood preparations. Aliquots of each sample were combined with five different platelet concentrations. In 46 of these blood preparations, mtDNAcn was measured by qPCR. MtDNAcn(WB) increased 1.07 (95%CI 0.86, 1.29; p<0.001) per 1000 platelets present in the preparation. We proved that leukocyte count should also be taken into account as mtDNAcn(WB) was inversely associated with leukocyte count; it increased 1.10 (95%CI 0.95, 1.25, p<0.001) per unit increase of the ratio between platelet and leukocyte counts. If hematological measurements are available, subtracting 1.10 the platelets/leukocyte ratio from mtDNAcn(WB) may serve as an estimation for mtDNAcn(L). Both platelet and leukocyte counts in the sample are important sources of variation if comparing mtDNAcn among groups of patients when mtDNAcn is measured in DNA extracted from whole blood. Not taking the platelet/leukocyte ratio into account in whole blood measurements, may lead to overestimation and misclassification if interpreted as leukocytes' mtDNAcn. PMID:27736919

  13. Leukocytes and Perfusion Recovery after Arterial Occlusion

    NARCIS (Netherlands)

    Haverslag, R.T.

    2012-01-01

    The main topic of this thesis is arteriogenesis and the role circulating leukocytes play in this process of perfusion recovery. We have used clinically applicable inhibitors to increase leukocyte attraction and extravasation and discovered previously unknown factors which play a role in

  14. Leukocyte telomere dynamics in the elderly

    DEFF Research Database (Denmark)

    Steenstrup, Troels; Hjelmborg, Jacob V B; Mortensen, Laust H;

    2013-01-01

    Limited data suggest that leukocytes of the elderly display ultra-short telomeres. It was reported that in some elderly persons leukocyte telomere length (LTL) shows age-dependent elongation. Using cross-sectional and longitudinal models, we characterized LTL dynamics in participants...

  15. Interactions between stably rolling leukocytes in vivo

    Science.gov (United States)

    King, Michael R.; Ruscio, Aimee D.; Kim, Michael B.; Sarelius, Ingrid H.

    2005-03-01

    We have characterized the two-dimensional spatial dependence of the hydrodynamic interactions between two adhesively rolling leukocytes in a live venule in the mouse cremaster muscle. Two rolling leukocytes were observed to slow each other down when rolling together in close proximity due to mutual sheltering from the external blood flow in the vessel lumen. A previous study of leukocyte rolling interactions using carbohydrate-coated beads in a parallel-plate flow chamber and a detailed computer model of adhesion in a multicellular environment is in qualitative agreement with the current in vivo results.

  16. Metabolism of phospholipids by polymorphonuclear leukocytes

    NARCIS (Netherlands)

    Elsbach, P.; Berg, J.W.O. van den; Bosch, H. van den; Deenen, L.L.M. van

    1965-01-01

    By incubating homogenates of polymorphonuclear leukocytes obtained from rabbit-peritoneal exudates with various 32P-labeled phosphoglycerides the following reactions were identified: Lecithin → Lysolecithin and fatty acid (1) Lysolecithin → Glycerylphosphorylcholine and fatty acid (2) Lysolecithin →

  17. The multiple faces of leukocyte interstitial migration

    Science.gov (United States)

    Lämmermann, Tim; Germain, Ronald N.

    2014-01-01

    Spatiotemporal control of leukocyte dynamics within tissues is critical for successful innate and adaptive immune responses. Homeostatic trafficking and coordinated infiltration into and within sites of inflammation and infection rely on signaling in response to extracellular cues that in turn controls a variety of intracellular protein networks regulating leukocyte motility, migration, chemotaxis, positioning, and cell–cell interaction. In contrast to mesenchymal cells, leukocytes migrate in an amoeboid fashion by rapid cycles of actin polymerization and actomyosin contraction, and their migration in tissues is generally referred to as low adhesive and nonproteolytic. The interplay of actin network expansion, contraction, and adhesion shapes the exact mode of amoeboid migration, and in this review, we explore how leukocyte subsets potentially harness the same basic biomechanical mechanisms in a cell-type-specific manner. Most of our detailed understanding of these processes derives from in vitro migration studies in three-dimensional gels and confined spaces that mimic geometrical aspects of physiological tissues. We summarize these in vitro results and then critically compare them to data from intravital imaging of leukocyte interstitial migration in mouse tissues. We outline the technical challenges of obtaining conclusive mechanistic results from intravital studies, discuss leukocyte migration strategies in vivo, and present examples of mode switching during physiological interstitial migration. These findings are also placed in the context of leukocyte migration defects in primary immunodeficiencies. This overview of both in vitro and in vivo studies highlights recent progress in understanding the molecular and biophysical mechanisms that shape robust leukocyte migration responses in physiologically complex and heterogeneous environments. PMID:24573488

  18. Effects of ochratoxin a on broiler leukocytes

    Directory of Open Access Journals (Sweden)

    MA Moura

    2004-09-01

    Full Text Available This study evaluated alterations in the qualitative cellular profile of leukocytes caused by the administration of low doses of ochratoxin-A (OTA in poultry. Sixty chicks were separated in three experimental groups: control, PBS-treated and OTA-treated. Blood smears from all birds were analyzed three and six hours post-treatment. Differential leukocyte counting demonstrated that OTA reduced the percentage of lymphocytes and eosinophils and significantly increased the number of heterophils and monocytes.

  19. Extração de DNA a partir de sangue humano coagulado para aplicação nas técnicas de genotipagem de antígenos leucocitários humanos e de receptores semelhantes à imunoglobulina DNA extraction from coagulated human blood for application in genotyping techniques for human leukocyte antigen and immunoglobulin-like receptors

    Directory of Open Access Journals (Sweden)

    Daniela Maira Cardozo

    2009-12-01

    Full Text Available O objetivo deste estudo foi padronizar uma metodologia de extração de DNA de alta qualidade a partir de amostras de sangue coagulado. Quarenta e oito amostras de sangue humano coagulado foram utilizadas para a extração de DNA pelo kit comercial EZ-DNA® (Biological Industries, Beit Haemek, Israel, pelo kit de coluna Neoscience® (One Lambda Inc., San Diego, CA e pelo método modificado de salting out. Apenas o método de salting out foi capaz de extrair altas concentrações de DNA (média, 180ng/µL, as quais foram medidas pelo detector de fluorescência Qubit® (Invitrogen, USA. Este método permitiu a amplificação dos genes HLA (human leukocyte antigens pela tecnologia PCR-SSO (polymerase chain reaction - specific sequence of oligonucleotides Luminex, a qual exige DNA de boa qualidade, e de genes KIR (killer cell immunoglobulin-like receptors pela técnica made in house PCR-SSP (polymerase chain reaction-sequence specific of primers, a qual demanda uma concentração específica de DNA (10ng/µL. Concluímos que a técnica de salting out modificada foi muito eficiente, simples e rápida para a extração de DNA de amostras de sangue humano coagulado, com o objetivo de realizar a genotipagem de genes HLA e KIR.The objective of this study was to standardize a method for extracting high-quality DNA from samples of coagulated blood. Forty-eight samples of human coagulated blood were used for DNA extraction by means of the EZ-DNA® commercial kit (Biological Industries, Beit Haemek, Israel, the Neoscience® column kit (One Lambda Inc., San Diego, CA, USA and a modified salting-out method. Only the salting-out method was able to extract high concentrations of DNA (mean, 180 ng/¼l, which were measured using the Qubit® fluorescence detector (Invitrogen, USA. This method enabled amplification of HLA (human leukocyte antigen genes using the Luminex PCR-SSO (polymerase chain reaction - sequence-specific oligonucleotide technology, which demands

  20. Plasmodium vivax: paroxysm-associated lipids mediate leukocyte aggregation

    Directory of Open Access Journals (Sweden)

    Mendis Kamini

    2007-05-01

    Full Text Available Abstract Background Paroxysms are recurrent febrile episodes, characteristic of Plasmodium vivax infections, which coincide with the rupture of schizont-infected erythrocytes in the patients' circulation. The present study describes the formation of prominent aggregates of leukocytes in vitro in the presence of parasite and host factors released during paroxysms. Methods Whole blood cells from uninfected malaria-naïve donors were incubated with plasma taken during a paroxysm or normal human plasma as a control and cell smears were observed under the microscope for the presence of leukocyte aggregates. Plasma factors involved in mediating the leukocyte aggregation were identified using immune depletion and reconstitution experiments. Furthermore, biochemical characterization was carried out to determine the chemical nature of the active moieties in plasma present during paroxysms. Results Leukocyte aggregates were seen exclusively when cells were incubated in plasma collected during a paroxysm. Immune depletion and reconstitution experiments revealed that the host cytokines TNF-alpha, GM-CSF, IL-6 and IL-10 and two lipid fractions of paroxysm plasma comprise the necessary and sufficient mediators of this phenomenon. The two lipid components of the paroxysm plasmas speculated to be of putative parasite origin, were a phospholipid-containing fraction and another containing cholesterol and triglycerides. The phospholipid fraction was dependent upon the presence of cytokines for its activity unlike the cholesterol/triglyceride-containing fraction which in the absence of added cytokines was much more active than the phospholipids fraction. The biological activity of the paroxysm plasmas from non-immune patients who presented with acute P. vivax infections was neutralized by immune sera raised against schizont extracts of either P. vivax or Plasmodium falciparum. However, immune sera against P. vivax were more effective than that against P. falciparum

  1. Leukocyte activation by triglyceride-rich lipoproteins.

    Science.gov (United States)

    Alipour, Arash; van Oostrom, Antonie J H H M; Izraeljan, Alisa; Verseyden, Caroline; Collins, Jennifer M; Frayn, Keith N; Plokker, Thijs W M; Elte, Jan Willem F; Castro Cabezas, Manuel

    2008-04-01

    Postprandial lipemia has been linked to atherosclerosis and inflammation. Because leukocyte activation is obligatory for atherogenesis, leukocyte activation by triglyceride-rich lipoproteins (TRLs) was investigated. The expression of CD11b and CD66b after incubation with glucose and native and artificial TRLs (NTRL and ATRL) in vivo and in vitro was evaluated by flowcytometry. Oral fat loading tests showed an increased expression of CD11b on monocytes and neutrophils and CD66b on neutrophils. In 11 volunteers, postprandial leukocytes became enriched with meal-derived fatty acids ([1-(13)C]16:0) suggesting uptake of exogenous fat. ApoB binding on leukocytes measured by flowcytometry in 65 subjects was highest on neutrophils and monocytes suggesting adherence of apoB-containing lipoproteins. Physiological concentrations of TRLs showed 62% increased neutrophil CD11b and a dose-dependent increased monocyte CD11b up to 84% in vitro. Incubations with lipid emulsions in the hypertriglyceridemic range showed a 5-fold increased monocyte CD11b expression, which was higher than the positive control (fMLP), and a dose-dependent 2- to 3-fold increased neutrophil CD11b and CD66b. The oxidative scavenger DMTU decreased the neutrophil CD66b expression by 36%. Acute hypertriglyceridemia is a leukocyte activator most likely by direct interaction between TRLs and leukocytes and uptake of fatty acids. TG-mediated leukocyte activation is an alternative proinflammatory and proatherogenic mechanism of hypertriglyceridemia in part associated to the generation of oxidative stress.

  2. Leukocyte margination in a model microvessel

    Science.gov (United States)

    Freund, Jonathan B.

    2007-02-01

    The physiological inflammation response depends upon the multibody interactions of blood cells in the microcirculation that bring leukocytes (white blood cells) to the vessel walls. We investigate the fluid mechanics of this using numerical simulations of 29 red blood cells and one leukocyte flowing in a two-dimensional microvessel, with the cells modeled as linearly elastic shell membranes. Despite its obvious simplifications, this model successfully reproduces the increasingly blunted velocity profiles and increased leukocyte margination observed at lower shear rates in actual microvessels. Red cell aggregation is shown to be unnecessary for margination. The relative stiffness of the red cells in our simulations is varied by over a factor of 10, but the margination is found to be much less correlated with this than it is to changes associated with the blunting of the mean velocity profile at lower shear rates. While velocity around the leukocyte when it is near the wall depends upon the red cell properties, it changes little for strongly versus weakly marginating cases. In the more strongly marginating cases, however, a red cell is frequently observed to be leaning on the upstream side of the leukocyte and appears to stabilize it, preventing other red cells from coming between it and the wall. A well-known feature of the microcirculation is a near-wall cell-free layer. In our simulations, it is observed that the leukocyte's most probable position is at the edge of this layer. This wall stand-off distance increases with velocity following a scaling that would be expected for a lubrication mechanism, assuming that there were a nearly constant force pushing the cells toward the wall. The leukocyte's near-wall position is observed to be less stable with increasing mean stand-off distance, but this distance would have potentially greater effect on adhesion since the range of the molecular binding is so short.

  3. Improvement of blood compatibility on polysulfone-polyvinylpyrrolidone blend films as a model membrane of dialyzer by physical adsorption of recombinant soluble human thrombomodulin (ART-123).

    Science.gov (United States)

    Omichi, Masaaki; Matsusaki, Michiya; Maruyama, Ikuro; Akashi, Mitsuru

    2012-01-01

    ART-123 is a recombinant soluble human thrombomodulin (hTM) with potent anticoagulant activity, and is available for developing antithrombogenic surfaces by immobilization. We focused on improving blood compatibility on the dialyzer surface by the physical adsorption of ART-123 as a safe yet simple method without using chemical reagents. The physical adsorption mechanism and anticoagulant activities of adsorbed hTM on the surface of a polysulfone (PSF) membrane containing polyvinylpyrrolidone (PVP) as a model dialyzer were investigated in detail. The PVP content of the PSF-PVP films was saturated at 20 wt% after immersion in Tris-HCl buffer, even with the addition of over 20 wt% PVP. The surface morphology of the PSF-PVP films was strongly influenced by the PVP content, because PVP covered the outermost surface of the PSF-PVP films. The adsorption speed of hTM slowed dramatically with increasing PVP content up to 10 wt%, but the maximum adsorption amount of hTM onto the PSF-PVP film surface was almost the same, regardless of the PVP content. The PSF-PVP film with the physically adsorbed hTM showed higher protein C activity as compared to the PSF film, it showed excellent blood compatibility due to the protein C activity and the inhibition properties of platelet adhesion. The physical adsorption of hTM can be useful as a safe yet simple method to improve the blood compatibility of a dialyzer surface.

  4. Cigarette smoking and leukocyte subpopulations in men.

    Science.gov (United States)

    Freedman, D S; Flanders, W D; Barboriak, J J; Malarcher, A M; Gates, L

    1996-07-01

    Because of previously reported associations among the total leukocyte count, cigarette smoking, and risk of cardiovascular disease, we examined the relation of cigarette smoking to various leukocyte subpopulations among 3467 men aged 31 to 45 years. The median total leukocyte count was 36% higher (7840 vs. 5760 cells/mL) among current cigarette smokers than among men who had never smoked, and both stratification and regression analyses were used to examine independent associations with leukocyte subpopulations. At equivalent counts of other subpopulations, CD4+ lymphocytes and neutrophils were the cell types most strongly associated with cigarette smoking; each standard deviation change in counts of these subpopulations increased the odds of current (vs. never) smoking by approximately threefold. Furthermore, whereas 15% of the 238 men with relatively low (men with relatively high counts of both subpopulations were current smokers. Counts of T lymphocytes also tended to be higher among the 32 men with self-reported ischemic heart disease than among other men. These results, along with previous reports of immunologically active T lymphocytes in atherosclerotic plaques, suggest that this subpopulation may be of particular interest in studies examining the relation of leukocytes to cardiovascular disease.

  5. Profile of anemia in chronic renal failure patients: comparison between predialyzed and dialyzed patients at the Division of Nephrology, Department of Internal Medicine, Sanglah Hospital, Denpasar, Bali, Indonesia.

    Science.gov (United States)

    Suega, Ketut; Bakta, Made; Dharmayudha, Tjok Gde; Lukman, Jodi S; Suwitra, Ketut

    2005-01-01

    To explore the profile of anemia in pradialytic and dialytic CRF patients at the Division of Nephrology, Department of Internal Medicine, Sanglah Hospital, Denpasar, from January to June 2000. There were 26 chronic dialyzed patients and 26 pradialyzed patients. Technicon H-1 was used to examine peripheral blood count; blood urea nitrogen and serum creatinine were examined using standard technique. MEIA (microparticle enzyme immunoassay) was used for serum folic acid and serum B12 level. Of 52 CRF patients, hemoglobin (Hb) levels ranged from 4.6 g/dl to 15.1 g/dl with a mean of 9.3 +/- 2.7 g/dl. Hematocrit levels ranged from 15.3% to 49.3%, with a mean of 29.8 +/- 7.9%. There was a significant difference in the hemoglobin and hematocrit levels of chronic dialyzed and pradialyzed patients. The prevalence of anemia (according to the WHO's criteria) among CRF patients was 84.5% (45/52), with the prevalence in chronic dialyzed patients being 100% and 73.1% in pradialyzed patients. If a hemoglobin level of less than 10 g/dl is used as a cut off point, the prevalence of anemia in dialyzed patients was 96.2% and 30.8% among pradialyzed patients. The severity of anemia among 26 dialyzed patients was: severe in 2 cases (8%), moderate in 16 cases (64%) and mild in 7 cases (28%), while in pradialytic patients the severity of anemia was: moderate in 4 patients (50%) and mild in 4 (50%). The morphology of 33 anemic patients was normocrhomic normocytic in 26 (78.8%) cases, slightly macrocytic in 7 (21.2%) cases, and no hypochromic anemia was found. Two anemic patients were associated with low serum folic acid (1 patient with macrocytic anemia and the other case with normochromic normocytic anemia). The serum B12 level was found to be normal in all cases. There was no significant correlation between the severity of anemia and serum creatinine levels or duration of hemodialysis. Anemia is a hallmark for CRF patients with a high prevalence of anemia and moderate degree of anemia

  6. Modeling leukocyte-leukocyte non-contact interactions in a lymph node.

    Directory of Open Access Journals (Sweden)

    Nicola Gritti

    Full Text Available The interaction among leukocytes is at the basis of the innate and adaptive immune-response and it is largely ascribed to direct cell-cell contacts. However, the exchange of a number of chemical stimuli (chemokines allows also non-contact interaction during the immunological response. We want here to evaluate the extent of the effect of the non-contact interactions on the observed leukocyte-leukocyte kinematics and their interaction duration. To this aim we adopt a simplified mean field description inspired by the Keller-Segel chemotaxis model, of which we report an analytical solution suited for slowly varying sources of chemokines. Since our focus is on the non-contact interactions, leukocyte-leukocyte contact interactions are simulated only by means of a space dependent friction coefficient of the cells. The analytical solution of the Keller-Segel model is then taken as the basis of numerical simulations of interactions between leukocytes and their duration. The mean field interaction force that we derive has a time-space separable form and depends on the chemotaxis sensitivity parameter as well as on the chemokines diffusion coefficient and their degradation rate. All these parameters affect the distribution of the interaction durations. We draw a successful qualitative comparison between simulated data and sets of experimental data for DC-NK cells interaction duration and other kinematic parameters. Remarkably, the predicted percentage of the leukocyte-leukocyte interactions falls in the experimental range and depends (~25% increase upon the chemotactic parameter indicating a non-negligible direct effect of the non-contact interaction on the leukocyte interactions.

  7. A liver perfusion model for studies of selective adherence and transient halting of portal blood leukocytes in sinusoids.

    Science.gov (United States)

    Durowicz, Sergiusz; Olszewski, Waldemar L

    2003-01-15

    Portal blood leukocytes play an important, but still poorly defined, role in the immune processes of the liver. In our previous studies, we showed that certain leukocyte subsets are selectively halted in the liver. These cells marginate in sinusoids and, together with resident Kupffer and endothelial sinusoidal cells, participate in antiviral and antitumor processes. The molecular mechanisms of margination and cooperation with resident sinusoidal cells require clarification. However, in vivo harvesting of portal blood leukocytes is associated with cumbersome cannulation of portal and hepatic veins and manipulation of the liver, causing major disturbances in splanchnic blood flow and liver blood supply, totally distorting sinusoidal blood perfusion and leukocyte margination. To overcome these difficulties, we have developed an in situ normothermic rat liver perfusion model permitting quantitative observations of blood leukocyte extraction in sinusoids. First, liver was flushed through the portal vein and the effluent leukocytes, named liver-associated leukocytes (LAL1), were collected from hepatic veins. Then, the liver was perfused for 60 min with 50 ml of blood using a semiclosed perfusion system. Upon completion of perfusion, the liver portal vasculature was flushed again to retrieve the leukocytes extracted from the perfusing blood (LAL2). These cells were characterized with respect to their phenotype and cytotoxicity. The mean leukocyte count of the washout before perfusion was 1.04+/-0.2x10(6)/g of liver tissue and 0.9+/-0.1x10(6)/g after 60 min of perfusion, indicating retention by the perfused liver, the live leukocyte extracting capacity. To further evaluate the efficiency of perfusion, FITC-labelled leukocytes were added to the perfusing leukocyte-free blood. Around 95% of the postperfusion washout LALs were FITC(+). Heat-killed leukocytes did not marginate in sinusoids. Preincubation of leukocytes with substances able to lower adhesion capacity, such as

  8. Epac inhibits apoptosis of human leukocytes

    NARCIS (Netherlands)

    Grandoch, M.; Bujok, V.; Fleckenstein, D.; Schmidt, M.; Fischer, J. W.; Weber, A. -A.

    2009-01-01

    cAMP is known to participate in the regulation of apoptosis in leukocytes. Depending on the cell type, pro- and antiapoptotic effects of cAMP have been described. Thus far, most of the cAMP-dependent effects have been attributed to the activation of PKA. However, Epac proteins (direct cAMP targets a

  9. Chemokines in the corpus luteum: Implications of leukocyte chemotaxis

    Directory of Open Access Journals (Sweden)

    Liptak Amy R

    2003-11-01

    Full Text Available Abstract Chemokines are small molecular weight peptides responsible for adhesion, activation, and recruitment of leukocytes into tissues. Leukocytes are thought to influence follicular atresia, ovulation, and luteal function. Many studies in recent years have focused attention on the characterization of leukocyte populations within the ovary, the importance of leukocyte-ovarian cell interactions, and more recently, the mechanisms of ovarian leukocyte recruitment. Information about the role of chemokines and leukocyte trafficking (chemotaxis during ovarian function is important to understanding paracrine-autocrine relationships shared between reproductive and immune systems. Recent advances regarding chemokine expression and leukocyte accumulation within the ovulatory follicle and the corpus luteum are the subject of this mini-review.

  10. Leukocyte set points in metabolic disease.

    Science.gov (United States)

    Odegaard, Justin I; Chawla, Ajay

    2012-01-01

    Vertebrate tissues comprise precise admixtures of parenchymal and hematopoietic cells, whose interactions are vital to proper tissue function. By regulating this interaction, vertebrates are able to mitigate environmental stress and coordinate dramatic physiologic adaptations. For instance, under conditions of chronic nutrient excess, leukocyte recruitment and activation increase in an effort to decrease excess nutrient storage and alleviate adipocyte stress. While basal equilibria may be reestablished upon normalization of nutrient intake, a new set point characterized by insulin resistance and chronic inflammation is established if the stress persists. Consequently, although this response is adaptive in settings of acute overfeeding and infection, it has catastrophic health consequences in the modern context of obesity. Understanding how leukocyte set points (numbers and activation status) are established, maintained, and regulated in tissues is, thus, critical to our understanding of, and intervention in, chronic metabolic diseases, such as obesity and diabetes.

  11. Indium-111 leukocyte scanning and fracture healing

    Energy Technology Data Exchange (ETDEWEB)

    Mead, L.P.; Scott, A.C.; Bondurant, F.J.; Browner, B.D. (Univ. of Texas Medical School, Houston (USA))

    1990-01-01

    This study was undertaken to determine the specificity of indium-111 leukocyte scans for osteomyelitis when fractures are present. Midshaft tibial osteotomies were performed in 14 New Zealand white rabbits, seven of which were infected postoperatively with Staphylococcus aureus per Norden's protocol. All 14 rabbits were scanned following injection with 75 microCi of indium 111 at 72 h after osteotomy and at weekly intervals for 4 weeks. Before the rabbits were killed, the fracture sites were cultured to document the presence or absence of infection. The results of all infected osteotomy sites were positive, whereas no positive scans were found in the noninfected osteotomies. We concluded from this study that uncomplicated fracture healing does not result in a positive indium-111 leukocyte scan.

  12. Loss of amino acids into dialysate during hemodialysis using hydrophilic and nonhydrophilic polyester-polymer alloy and polyacrylonitrile membrane dialyzers.

    Science.gov (United States)

    Yokomatsu, Atsuko; Fujikawa, Tetsuya; Toya, Yoshiyuki; Shino-Kakimoto, Midori; Itoh, Yoko; Mitsuhashi, Hiroshi; Tamura, Kouichi; Hirawa, Nobuhito; Yasuda, Gen; Umemura, Satoshi

    2014-08-01

    During hemodialysis, amino acid loss through the dialysate remained a significant problem and was not clear in some dialyzers; therefore, we investigated amino acid loss with hydrophilic and nonhydrophilic polyester-polymer alloy membranes and polyacrylonitrile membranes. Nine maintenance hemodialysis patients were studied to assess amino acid loss during hemodialysis with the three membranes. Total amino acid losses were 85.7 ± 27.2 mg/L, 83.3 ± 16.1 mg/L, and 72.1 ± 22.5 mg/L with the hydrophilic, nonhydrophilic polyester-polymer alloy, and polyacrylonitrile membranes, respectively. Amino acid losses were greater with the hydrophilic membrane compared with the polyacrylonitrile membrane for ornithine (2.0 ± 0.6 vs. 1.4 ± 0.4 mg/L, P = 0.025), phenylalanine (2.4 ± 0.9 vs. 1.8 ± 0.8 mg/L, P = 0.012), and tryptophan (0.6 ± 0.2 vs. 0.4 ± 0.2 mg/L, P = 0.023). Amino acid losses were greater with the nonhydrophilic membrane than with the polyacrylonitrile membrane for ornithine (2.0 ± 0.4 vs. 1.4 ± 0.4 mg/L, P = 0.017), phenylalanine (2.3 ± 0.5 vs. 1.8 ± 0.8 mg/L, P = 0.018), tryptophan (0.7 ± 0.2 vs. 0.4 ± 0.2 mg/L, P = 0.003), and cystine (3.2 ± 0.7 vs. 2.0 ± 0.7 mg/L, P = 0.005). In conclusion, greater losses of ornithine, phenylalanine, tryptophan, and cystine were observed with polyester-polymer alloy than with polyacrylonitrile membranes during hemodialysis. Constant attention should be paid to the amino acid loss profile to improve nutritional control in hemodialysis patients.

  13. Automated leukocyte recognition using fuzzy divergence.

    Science.gov (United States)

    Ghosh, Madhumala; Das, Devkumar; Chakraborty, Chandan; Ray, Ajoy K

    2010-10-01

    This paper aims at introducing an automated approach to leukocyte recognition using fuzzy divergence and modified thresholding techniques. The recognition is done through the segmentation of nuclei where Gamma, Gaussian and Cauchy type of fuzzy membership functions are studied for the image pixels. It is in fact found that Cauchy leads better segmentation as compared to others. In addition, image thresholding is modified for better recognition. Results are studied and discussed.

  14. Bovine leukocyte adhesion deficiency (BLAD): a review.

    Science.gov (United States)

    Nagahata, Hajime

    2004-12-01

    Bovine leukocyte adhesion deficiency (BLAD) in Holstein cattle is an autosomal recessive congenital disease characterized by recurrent bacterial infections, delayed wound healing and stunted growth, and is also associated with persistent marked neutrophilia. The molecular basis of BLAD is a single point mutation (adenine to guanine) at position 383 of the CD18 gene, which caused an aspartic acid to glycine substitution at amino acid 128 (D128G) in the adhesion molecule CD18. Neutrophils from BLAD cattle have impaired expression of the beta2 integrin (CD11a,b,c/CD18) of the leukocyte adhesion molecule. Abnormalities in a wide spectrum of adherence dependent functions of leukocytes have been fully characterized. Cattle affected with BLAD have severe ulcers on oral mucous membranes, severe periodontitis, loss of teeth, chronic pneumonia and recurrent or chronic diarrhea. Affected cattle die at an early age due to the infectious complications. Holstein bulls, including carrier sires that had a mutant BLAD gene in heterozygote were controlled from dairy cattle for a decade. The control of BLAD in Holstein cattle by publishing the genotypes and avoiding the mating between BLAD carriers was found to be successful. This paper provides an overview of the genetic disease BLAD with reference to the disease in Holstein cattle.

  15. Comparison of technetium-99m-HM-PAO leukocytes with indium-111-oxine leukocytes for localizing intraabdominal sepsis

    Energy Technology Data Exchange (ETDEWEB)

    Mountford, P.J.; Kettle, A.G.; O' Doherty, M.J.; Coakley, A.J. (Kent and Canterbury Hospital (England))

    1990-03-01

    Technetium-99m-HM-PAO (({sup 99m}Tc)HM-PAO) leukocyte and indium-111-oxine (111In-oxine) leukocyte scanning were carried out simultaneously in 41 patients at 4 hr and 24 hr after reinjection to determine whether the 4-hr {sup 99m}Tc scan could replace the 24-hr {sup 111}In scan for detecting intraabdominal sepsis. Abdominal infection was confirmed in 12 cases. The 4-hr {sup 99}Tc-leukocyte scan, the 4-hr {sup 111}In-leukocyte scan, and the 24-hr {sup 111}In-leukocyte scan yielded a sensitivity of 100%, 67%, and 100%, respectively, and a specificity of 62%, 90%, and 86%, respectively. The 24-hr {sup 99m}Tc-leukocyte scan also produced a sensitivity of 100%, but it was falsely positive in all 29 cases without infection due to physiologic bowel uptake. False-positive 4-hr {sup 99m}Tc-leukocyte scans were also produced by physiologic bowel uptake in seven cases all of whom had true-negative 4-hr and 24-hr {sup 111}In-leukocyte scans. Because of the high incidence of false-positive 4-hr ({sup 99m}Tc)HM-PAO leukocyte scans, it was concluded that they could not replace 24-hr {sup 111}In-leukocyte scans for detecting intraabdominal sepsis, and that serial {sup 99m}Tc leukocyte scans starting earlier than 4 hr after reinjection must be evaluated.

  16. Impact of human leukocyte antigen mismatching on outcomes of liver transplantation:A meta-analysis

    Institute of Scientific and Technical Information of China (English)

    2010-01-01

    AIM:To assess the effect of human leukocyte antigen(HLA) mismatching on liver graft outcome and acute rejection from a meta-analysis of available cohort studies.METHODS:Articles in PubMed/MEDLINE,EMBASE and the Cochrane database from January 1970 to June 2009,including non-English literature identified in these databases,were searched.Only studies comparing HLA or sub-phenotype matching with mismatching were extracted.The percentage of graft survival was extracted by "Engauge Digitizer" from survival curves...

  17. Killing of gram-negative bacteria by polymorphonuclear leukocytes: role of an O2-independent bactericidal system.

    Science.gov (United States)

    Weiss, J; Victor, M; Stendhal, O; Elsbach, P

    1982-04-01

    Previous studies have suggested that a cationic bactericidal/permeability-increasing protein (BPI) present in both rabbit and human polymorphonuclear leukocytes is the principal O2-independent bactericidal agent of these cells toward several strains of Escherichia coli and Salmonella typhimurium (1978. J. Biol. Chem. 253: 2664--2672; 1979. J. Biol. Chem. 254: 11000--11009). To further evaluate the possible role of this protein in the killing of gram-negative bacteria by polymorphonuclear leukocytes, we have measured the bactericidal activity of intact rabbit peritoneal exudate leukocytes under aerobic or anaerobic conditions and of intact human leukocytes from a patient with chronic granulomatous disease. Anaerobic conditions were created by flushing the cells under a nitrogen stream. Effective removal of oxygen was demonstrated by the inability of nitrogen-flushed leukocytes to mount a respiratory burst (measured as increased conversion of 1-[14C]glucose leads to 14CO2 or by superoxide production) during bacterial ingestion. At a bacteria/leukocyte ratio of 10:1, killing of gram-positive, BPI-resistant, Staphylococcus epidermidis is markedly impaired in the absence of oxygen (76.4 +/- 3.3% killing in room air, 29.2 +/- 8.2% killing in nitrogen). Essentially all increased bacterial survival is intracellular. In contrast, both a nonopsonized rough strain (MR-10) and an opsonized smooth strain (MS) of S. typhimurium 395 are killed equally well in room air and nitrogen. A maximum of 70--80 MR-10 and 30--40 MS are killed per leukocyte either in the presence or absence of oxygen. There is no intracellular bacterial survival in either condition indicating that intracellular O2-independent bactericidal system(s) of rabbit polymorphonuclear leukocytes can at least match the leukocyte's ingestive capacity. Whole homogenates and crude acid extracts manifest similar bactericidal capacity toward S. typhimurium 395. This activity can be accounted for by the BPI content of these

  18. Flow and deformation of the capillary glycocalyx in the wake of a leukocyte

    Science.gov (United States)

    Damiano, Edward R.; Stace, Thomas M.

    2005-03-01

    An analysis is presented of the axisymmetric axial and radial flow and deformation fields throughout the endothelial-cell glycocalyx surface layer in the wake region behind a leukocyte moving steadily through a capillary. The glycocalyx, modeled as a thin poroelastic surface layer lining the capillary wall, is assumed to consist of a binary mixture of a linearly viscous fluid constituent and an isotropic, highly compressible, linearly elastic solid constituent having a vanishingly small solid-volume fraction. Invoking the asymptotic approximations of lubrication theory in a frame of reference translating with the leukocyte, closed-form solutions are obtained to the leading-order boundary-value problems governing the axial and radial flow and deformation fields throughout the glycocalyx as well as the axial and radial flow fields throughout the free capillary lumen within the wake. A simple asymptotic expression is obtained for the length lchar of the wake region in terms of the translational speed U0 of the leukocyte, and the equilibrium thickness h0, permeability k0, and aggregate elastic modulus HA of the glycocalyx. The predicted wake length, as seen from an observer moving in a reference frame attached to the leukocyte, is consistent with the recovery time predicted from a one-dimensional analysis of glycocalyx deformation through a quiescent inviscid fluid. The two-dimensional fluid dynamical analysis presented here thus provides the appropriate relationships for extracting estimates of the mechanoelectrochemical properties of the glycocalyx from physiologically realistic constitutive models developed under simplified one-dimensional flow regimes. The directly measurable quantities lchar,U0, and h0, which are obtainable from in vivo observations of the wake region behind a leukocyte moving steadily through a capillary, can therefore be connected, through the results of this analysis, to estimates of the mechanoelectrochemical properties of the glycocalyx on

  19. Classification of human leukocyte antigen (HLA) supertypes

    DEFF Research Database (Denmark)

    Wang, Mingjun; Claesson, Mogens H

    2014-01-01

    Identification of new antigenic peptides, derived from infectious agents or cancer cells, which bind to human leukocyte antigen (HLA) class I and II molecules, is of importance for the development of new effective vaccines capable of activating the cellular arm of the immune response. However...... this complexity is to group thousands of different HLA molecules into several so-called HLA supertypes: a classification that refers to a group of HLA alleles with largely overlapping peptide binding specificities. In this chapter, we focus on the state-of-the-art classification of HLA supertypes including HLA...

  20. Analysis of leukocyte rolling in vivo and in vitro.

    Science.gov (United States)

    Sperandio, Markus; Pickard, John; Unnikrishnan, Sunil; Acton, Scott T; Ley, Klaus

    2006-01-01

    Leukocyte rolling is an important step for the successful recruitment of leukocytes from blood to tissues mediated by a specialized group of glycoproteins termed selectins. Because of the dynamic process of leukocyte rolling, binding of selectins to their respective counter-receptors (selectin ligands) needs to fulfill three major requirements: (1) rapid bond formation, (2) high tensile strength, and (3) fast dissociation rates. These criteria are perfectly met by selectins, which interact with specific carbohydrate determinants on selectin ligands. This chapter describes the theoretical background, technical requirements, and analytical tools needed to quantitatively assess leukocyte rolling in vivo and in vitro. For the in vivo setting, intravital microscopy allows the observation and recording of leukocyte rolling under different physiological and pathological conditions in almost every organ. Real-time and off-line analysis tools help to assess geometric, hemodynamic, and rolling parameters. Under in vitro conditions, flow chamber assays such as parallel plate flow chamber systems have been the mainstay to study interactions between leukocytes and adhesion molecules under flow. In this setting, adhesion molecules are immobilized on plastic, in a lipid monolayer, or presented on cultured endothelial cells on the chamber surface. Microflow chambers are available for studying leukocyte adhesion in the context of whole blood and without blood cell isolation. The microscopic observation of leukocyte rolling in different in vivo and in vitro settings has significantly contributed to our understanding of the molecular mechanisms responsible for the stepwise extravasation of leukocytes into inflamed tissues.

  1. Toward the treatment for Alzheimer's disease: adsorption is primary mechanism of removing amyloid β protein with hollow-fiber dialyzers of the suitable materials, polysulfone and polymethyl methacrylate.

    Science.gov (United States)

    Kawaguchi, Kazunori; Saigusa, Akira; Yamada, Shinji; Gotoh, Takehiro; Nakai, Shigeru; Hiki, Yoshiyuki; Hasegawa, Midori; Yuzawa, Yukio; Kitaguchi, Nobuya

    2016-06-01

    The accumulation of amyloid β protein (Aβ) in the brain reflects cognitive impairment in Alzheimer's disease. We hypothesized that the rapid removal of Aβ from the blood by an extracorporeal system may act as a peripheral Aβ sink from the brain. The present study aimed to determine the optimal materials and modality for Aβ removal by hemodialyzers. In a batch analysis, hollow-fiber fragments of polysulfone (PSf) and polymethyl methacrylate (PMMA) showed greater removal efficiency of Aβ than did other materials, such as cellulose-triacetates and ethylene-vinyl alcohol copolymer (PSf:PMMA at 30 min, 98.6 ± 2.4 %:97.8 ± 0.4 % for Aβ1-40 and 96.6 ± 0.3 %:99.0 ± 1.0 % for Aβ1-42). In a modality study, the Aβ solution was applied to PSf dialyzers and circulated in the dialysis and Air-filled adsorption-mode (i.e., the outer space of the hollow fibers was filled with air) or phosphate-buffered saline (PBS)-filled adsorption modes. The Aβ1-40 removal efficiency of the pre/post dialyzer in the Air-filled adsorption-mode was the highest (62.4 ± 12.6 %, p = 0.007). In a flow rate study in the Air-filled adsorption-mode, 200 mL/min showed the highest Aβ1-40 reduction rate of pool solution (97.3 ± 0.8 % at 15 min) compared with 20 mL/min (p = 0.00001) and 50 mL/min (p = 0.00382). PMMA dialyzers showed similar high reduction rates. Thus, the optimal modality for Aβ removal was the adsorption-mode with PSf or PMMA hollow fibers at around 50 mL/min flow rate, which seems to be suitable for clinical use.

  2. Competitive evaluation of data mining algorithms for use in classification of leukocyte subtypes with Raman microspectroscopy.

    Science.gov (United States)

    Maguire, A; Vega-Carrascal, I; Bryant, J; White, L; Howe, O; Lyng, F M; Meade, A D

    2015-04-07

    Raman microspectroscopy has been investigated for some time for use in label-free cell sorting devices. These approaches require coupling of the Raman spectrometer to complex data mining algorithms for identification of cellular subtypes such as the leukocyte subpopulations of lymphocytes and monocytes. In this study, three distinct multivariate classification approaches, (PCA-LDA, SVMs and Random Forests) are developed and tested on their ability to classify the cellular subtype in extracted peripheral blood mononuclear cells (T-cell lymphocytes from myeloid cells), and are evaluated in terms of their respective classification performance. A strategy for optimisation of each of the classification algorithm is presented with emphasis on reduction of model complexity in each of the algorithms. The relative classification performance and performance characteristics are highlighted, overall suggesting the radial basis function SVM as a robust option for classification of leukocytes with Raman microspectroscopy.

  3. Two New Monoterpene Glycosides from Qing Shan Lu Shui Tea with Inhibitory Effects on Leukocyte-Type 12-Lipoxygenase Activity

    Directory of Open Access Journals (Sweden)

    Ding Zhi Fang

    2013-04-01

    Full Text Available We evaluated the inhibitory effect of 12 Chinese teas on leukocyte-type 12-lipoxygenase (LOX activity. Tea catechins such as epigallocatechin gallate have been known to exhibit leukocyte-type 12-LOX inhibition. Qing Shan Lu Shui, which contains lower catechin levels than the other tested teas, suppressed leukocyte-type 12-LOX activity. To characterize the bioactive components of Qing Shan Lu Shui, leukocyte-type 12-LOX inhibitory activity–guided fractionation of the aqueous ethanol extract of the tea was performed, resulting in the isolation of two new monoterpene glycosides: liguroside A (1 and B (2. The structures of compounds 1 and 2 were characterized as (2E,5E-7-hydroperoxy-3,7-dimethyl-2,5-octadienyl-O-(α-L-rhamnopyranosyl-(1″→3′-(4′″-O-trans-p-coumaroyl-β-D-glucopyranoside and (2E,5E-7-hydroperoxy-3,7-dimethyl-2,5-octa-dienyl- O-(α-L-rhamnopyranosyl-(1″→3′-(4′″-O-cis-p-coumaroyl-β-D-glucopyranoside, respectively, based on spectral and chemical evidence. Ligurosides A (1 and B (2 showed inhibitory effects on leukocyte-type 12-LOX activity, with IC50 values of 1.7 and 0.7 μM, respectively.

  4. Effect of fatty acids on leukocyte function

    Directory of Open Access Journals (Sweden)

    Pompéia C.

    2000-01-01

    Full Text Available Fatty acids have various effects on immune and inflammatory responses, acting as intracellular and intercellular mediators. Polyunsaturated fatty acids (PUFAs of the omega-3 family have overall suppressive effects, inhibiting lymphocyte proliferation, antibody and cytokine production, adhesion molecule expression, natural killer cell activity and triggering cell death. The omega-6 PUFAs have both inhibitory and stimulatory effects. The most studied of these is arachidonic acid that can be oxidized to eicosanoids, such as prostaglandins, leukotrienes and thromboxanes, all of which are potent mediators of inflammation. Nevertheless, it has been found that many of the effects of PUFA on immune and inflammatory responses are not dependent on eicosanoid generation. Fatty acids have also been found to modulate phagocytosis, reactive oxygen species production, cytokine production and leukocyte migration, also interfering with antigen presentation by macrophages. The importance of fatty acids in immune function has been corroborated by many clinical trials in which patients show improvement when submitted to fatty acid supplementation. Several mechanisms have been proposed to explain fatty acid modulation of immune response, such as changes in membrane fluidity and signal transduction pathways, regulation of gene transcription, protein acylation, and calcium release. In this review, evidence is presented to support the proposition that changes in cell metabolism also play an important role in the effect of fatty acids on leukocyte functioning, as fatty acids regulate glucose and glutamine metabolism and mitochondrial depolarization.

  5. Optimization and pharmacological validation of a leukocyte migration assay in zebrafish larvae for the rapid in vivo bioactivity analysis of anti-inflammatory secondary metabolites.

    Directory of Open Access Journals (Sweden)

    María Lorena Cordero-Maldonado

    Full Text Available Over the past decade, zebrafish (Danio rerio have emerged as an attractive model for in vivo drug discovery. In this study, we explore the suitability of zebrafish larvae to rapidly evaluate the anti-inflammatory activity of natural products (NPs and medicinal plants used in traditional medicine for the treatment of inflammatory disorders. First, we optimized a zebrafish assay for leukocyte migration. Inflammation was induced in four days post-fertilization (dpf zebrafish larvae by tail transection and co-incubation with bacterial lipopolysaccharides (LPS, resulting in a robust recruitment of leukocytes to the zone of injury. Migrating zebrafish leukocytes were detected in situ by myeloperoxidase (MPO staining, and anti-inflammatory activity was semi-quantitatively scored using a standardized scale of relative leukocyte migration (RLM. Pharmacological validation of this optimized assay was performed with a panel of anti-inflammatory drugs, demonstrating a concentration-responsive inhibition of leukocyte migration for both steroidal and non-steroidal anti-inflammatory drugs (SAIDs and NSAIDs. Subsequently, we evaluated the bioactivity of structurally diverse NPs with well-documented anti-inflammatory properties. Finally, we further used this zebrafish-based assay to quantify the anti-inflammatory activity in the aqueous and methanolic extracts of several medicinal plants. Our results indicate the suitability of this LPS-enhanced leukocyte migration assay in zebrafish larvae as a front-line screening platform in NP discovery, including for the bioassay-guided isolation of anti-inflammatory secondary metabolites from complex NP extracts.

  6. Total antioxidant capacity, total phenolic content and iron and zinc dialyzability in selected Greek varieties of table olives, tomatoes and legumes from conventional and organic farming.

    Science.gov (United States)

    Drakou, Marina; Birmpa, Angeliki; Koutelidakis, Antonios E; Komaitis, Michael; Panagou, Efstathios Z; Kapsokefalou, Maria

    2015-03-01

    The objective was to compare 10 types of table olives, 11 types of tomatoes and tomato products and 18 types of legumes from conventional or organic farming for selected nutritional properties. All products were tested for their total antioxidant capacity (TAC) (Ferric Reducing Antioxidant Power (FRAP) assay) and total phenolic content (Folin-Ciocalteau method). Tomatoes and legumes were further tested for iron and zinc dialyzability after in vitro digestion. Ascorbic acid content of tomatoes was also measured. The study resulted that the nutritional properties of olives, tomatoes and legumes tested were different among the various cultivars but, in most cases, not between products from organic or conventional farming. Natural black olives, cherry and santorini tomatoes and lentils exhibited superior nutritional properties.

  7. Effectiveness of Haemodiafiltration with Heat Sterilized High-Flux Polyphenylene HF Dialyzer in Reducing Free Light Chains in Patients with Myeloma Cast Nephropathy.

    Directory of Open Access Journals (Sweden)

    Mathieu Rousseau-Gagnon

    Full Text Available In cases of myeloma cast nephropathy in need of haemodialysis (HD, reduction of free light chains using HD with High-Cut-Off filters (HCO-HD, in combination with chemotherapy, may be associated with better renal recovery. The aim of the present study is to evaluate the effectiveness of haemodiafiltration (HDF in reducing free light chain levels using a less expensive heat sterilized high-flux polyphenylene HF dialyzer (HF-HDF.In a single-centre prospective cohort study, 327 dialysis sessions were performed using a 2.2 m2 heat sterilized high-flux polyphenylene HF dialyzer (Phylther HF22SD, a small (1.1m2 or large (2.1 m2 high-cut-off (HCO dialyzer (HCOS and HCOL in a cohort of 16 patients presenting with dialysis-dependent acute cast nephropathy and elevated free light chains (10 kappa, 6 lambda. The outcomes of the study were the mean reduction ratio (RR of kappa and lambda, the proportion of treatments with an RR of at least 0.65, albumin loss and the description of patient outcomes. Statistical analysis was performed using linear and logistic regression through generalized estimating equation analysis so as to take into account repeated observation within subjects and adjust for session duration.There were no significant differences in the estimated marginal mean of kappa RR, which were respectively 0.67, 0.69 and 0.70 with HCOL-HD, HCOS-HDF and HF-HDF (P = 0.950. The estimated marginal mean of the proportions of treatments with a kappa RR ≥0.65 were 68%, 63% and 71% with HCOL-HD, HCOS-HDF and HF-HDF, respectively (P = 0.913. The estimated marginal mean of lambda RR were higher with HCOL-HDF (0.78, compared to HCOL-HD and HF-HDF (0.62, and 0.61 respectively. The estimated marginal mean proportion of treatments with a lambda RR ≥0.65 were higher with HCOL-HDF (81%, compared to 57% in HF-HDF (P = 0.042. The median albumin loss were 7, 21 and 63 g/session with HF-HDF, HCOL-HD and HCOL-HDF respectively (P = 0.044. Among survivors, 9 out of 10

  8. 聚偏氟乙烯中空纤维血液透析器的研究%Study of polyvinylidene fluoride hollow fiber dialyzers

    Institute of Scientific and Technical Information of China (English)

    张娟; 郭建辉; 吕晓龙; 武春瑞; 贾悦; 王暄; 陈华艳; 高启君

    2011-01-01

    目的 目前,用于血液透析的膜材料主要有聚砜和聚醚砜,在去除β2-微球蛋白及生物相容性方面尚存在缺陷.聚偏氟乙烯(PVDF)在血液透析方面尚无应用,本研究对PVDF膜材料进行了力学性能及血液透析性能的评价.方法 采用干-湿相转化法制备PVDF中空纤维膜;采用离心浇铸法将中空纤维膜封装成血液透析器,用实验室配制的模拟液代替人体血液进行血液透析实验.结果 PVDF膜断裂强力为20 cN左右,拉伸伸长率为200%左右;纤维壁厚为30μm的透析器对尿素和溶菌酶的清除率分别为90%和75%,对牛血清白蛋白的截留率可达到90%以上;添加剂聚乙二醇(PEG)质量分数为19%条件下牛血清白蛋白的截留率为91%,较其质量分数为22%的条件下截留率高;增大透析液流速,可以提高尿素和溶菌酶的清除率,但对牛血清白蛋白的截留率影响不大.结论 膜的力学性能满足透析要求,对尿素及溶菌酶的清除率有很大提高,对牛血清白蛋白的截留率比较理想,可以作为透析膜材料进一步开发利用.%Objective Currently applied membrane materials for hemodialysis are mainly polysulfone and polyethers. However, defects still remain in the removal of β2-MG and biocompatibility. Polyvinylidene fluoride (PVDF) has not been put in use in hemodialysis. This study focused on the evaluation of mechanics performance and the dialysis performance of PVDF membrane material. Methods PVDF hollow fiber membranes were spun by changing the membrane forming conditions. The membranes were sealed to dialyzers by centrifugal moulding method. The simulation solution, instead of patient blood, was used to evaluate the dialysis performance of PVDF hollow fiber membrane dialyzers. Results The tensile failure strength of PVDF membrane was about 20 cN. The tensile elongation of PVDF membrane was about 200%. The clearance rates of the dialyzers prepared with the fiber whose wall thickness was 30

  9. Effects of Pneumolysin on Human Polymorphonuclear Leukocytes and Platelets

    OpenAIRE

    Johnson, M K; Boese-Marrazzo, D; Pierce, W. A.

    1982-01-01

    Pneumolysin was bound by human polymorphonuclear leukocytes in a reaction which occurred very rapidly at 0 degrees C. Low concentrations of pneumolysin were found to stimulate leukocyte migration and lysosomal enzyme secretion. At increasing lysin levels, inhibition of spontaneous migration and chemotaxis, cell death, and lysis were observed. Pneumolysin was also found to lyse platelets and to activate serum to become chemotactic.

  10. 21 CFR 864.7675 - Leukocyte peroxidase test.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Leukocyte peroxidase test. 864.7675 Section 864... peroxidase test. (a) Identification. A leukocyte peroxidase test is a device used to distinguish certain... peroxidase activity as evidenced by staining. The results of this test are used in the differential...

  11. The effects of stress on the enzymes of peripheral leukocytes

    Science.gov (United States)

    Leise, E. M.; Gray, I.

    1973-01-01

    Previous work showed an early response of rabbit and human leukocyte enzymes to the stress of bacterial infection. Since these represented a mixed population of leukocytes and since polymorphonuclear leukocytes (PMN) increased in these preparations, it was necessary to establish whether the observed increase in lactate dehydrenase (LDH) and protein was the result of an increase in any one particular cell type or in all cells. The need for the development of a simple reproducible method for the differential separation of peripheral leukocytes for the furtherance of our own studies was apparent. It was also becoming increasingly apparent that morphologically similar cells, such as small lymphocytes (L) and macrophages, were capable of different biological functions. A dextran gradient centrifugation method was developed which has provided an easily reproducible technique for separating L from PMN. During the course of this work, in which over 250 rabbits were examined, the pattern of daily leukocyte protein and enzyme variation became increasingly more apparent. This information could have some impact on future work with leukocyte enzymes, by our group and by other workers. The differences in normal protein and enzyme levels maintained by some individuals, and some inbred strains, were evaluated and reported separately. It has been shown that one type of leukocyte may react more to a given stress than other leukocytes.

  12. Asymmetric membrane filters for the removal of leukocytes from blood

    NARCIS (Netherlands)

    Bruil, A.; Aken, van W.G.; Beugeling, T.; Feijen, J.; Steneker, I.; Huisman, J.G.; Prins, H.K.

    1991-01-01

    As part of a study on the mechanisms of leukocyte filtration, the influence of pore size distribution on filter efficiency was investigated. Conventional leukocyte filters are not suitable for model studies, as these filters are composed of tightly packed synthetic fibers, with a poorly defined poro

  13. File list: His.Bld.05.AllAg.Polymorphonuclear_leukocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Bld.05.AllAg.Polymorphonuclear_leukocytes hg19 Histone Blood Polymorphonuclear ...leukocytes http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/His.Bld.05.AllAg.Polymorphonuclear_leukocytes.bed ...

  14. File list: Unc.Bld.50.AllAg.Polymorphonuclear_leukocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Unc.Bld.50.AllAg.Polymorphonuclear_leukocytes hg19 Unclassified Blood Polymorphonuc...lear leukocytes http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/Unc.Bld.50.AllAg.Polymorphonuclear_leukocytes.bed ...

  15. File list: His.Bld.10.AllAg.Polymorphonuclear_leukocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Bld.10.AllAg.Polymorphonuclear_leukocytes hg19 Histone Blood Polymorphonuclear ...leukocytes http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/His.Bld.10.AllAg.Polymorphonuclear_leukocytes.bed ...

  16. File list: Unc.Bld.20.AllAg.Polymorphonuclear_leukocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Unc.Bld.20.AllAg.Polymorphonuclear_leukocytes hg19 Unclassified Blood Polymorphonuc...lear leukocytes http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/Unc.Bld.20.AllAg.Polymorphonuclear_leukocytes.bed ...

  17. File list: His.Bld.50.AllAg.Polymorphonuclear_leukocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Bld.50.AllAg.Polymorphonuclear_leukocytes hg19 Histone Blood Polymorphonuclear ...leukocytes http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/His.Bld.50.AllAg.Polymorphonuclear_leukocytes.bed ...

  18. File list: Unc.Bld.10.AllAg.Polymorphonuclear_leukocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Unc.Bld.10.AllAg.Polymorphonuclear_leukocytes hg19 Unclassified Blood Polymorphonuc...lear leukocytes http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/Unc.Bld.10.AllAg.Polymorphonuclear_leukocytes.bed ...

  19. File list: His.Bld.20.AllAg.Polymorphonuclear_leukocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Bld.20.AllAg.Polymorphonuclear_leukocytes hg19 Histone Blood Polymorphonuclear ...leukocytes http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/His.Bld.20.AllAg.Polymorphonuclear_leukocytes.bed ...

  20. File list: Unc.Bld.05.AllAg.Polymorphonuclear_leukocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Unc.Bld.05.AllAg.Polymorphonuclear_leukocytes hg19 Unclassified Blood Polymorphonuc...lear leukocytes http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/Unc.Bld.05.AllAg.Polymorphonuclear_leukocytes.bed ...

  1. Penetration of equine leukocytes by merozoites of Sarcocystis neurona.

    Science.gov (United States)

    Lindsay, David S; Mitchell, Sheila M; Yang, Jibing; Dubey, J P; Gogal, Robert M; Witonsky, Sharon G

    2006-06-15

    Horses are considered accidental hosts for Sarcocystis neurona and they often develop severe neurological disease when infected with this parasite. Schizont stages develop in the central nervous system (CNS) and cause the neurological lesions associated with equine protozoal myeloencephalitis. The present study was done to examine the ability of S. neurona merozoites to penetrate and develop in equine peripheral blood leukocytes. These infected host cells might serve as a possible transport mechanism into the CNS. S. neurona merozoites penetrated equine leukocytes within 5 min of co-culture. Infected leukocytes were usually monocytes. Infected leukocytes were present up to the final day of examination at 3 days. Up to three merozoites were present in an infected monocyte. No development to schizont stages was observed. All stages observed were in the host cell cytoplasm. We postulate that S. neurona merozoites may cross the blood brain barrier hidden inside leukocytes. Once inside the CNS these merozoites can egress and invade additional cells and cause encephalitis.

  2. Leukocytic oxygen activation and microbicidal oxidative toxins.

    Science.gov (United States)

    Hurst, J K; Barrette, W C

    1989-01-01

    Following a brief introduction of cellular response to stimulation comprising leukocyte activation, three major areas are discussed: (1) the neutrophil oxidase; (2) myeloperoxidase (MPO)-dependent oxidative microbicidal reactions; and (3) MPO-independent oxidative reactions. Topics included in section (A) are current views on the activation mechanism, redox composition, structural and topographic organization of the oxidase, and its respiratory products. In section (B), emphasis is placed on recent research on cidal mechanisms of HOCl, including the oxidative biochemistry of active chlorine compounds, identification of sites of lesions in bacteria, and attendant metabolic consequences. In section (C), we review the (bio)chemistry of H2O2 and .OH microbicidal reactions, with particular attention being given to addressing the controversial issue of probe methods to identify .OH radical and critical assessment of the recent proposal that MPO-independent killing arises from site-specific metal-catalyzed Fenton-type chemistry.

  3. Plasminogen activator inhibitor-2 (PAI-2) in eosinophilic leukocytes.

    Science.gov (United States)

    Swartz, Jonathan M; Byström, Jonas; Dyer, Kimberly D; Nitto, Takeaki; Wynn, Thomas A; Rosenberg, Helene F

    2004-10-01

    Plasminogen activator inhibitor-2 (PAI-2) as a potential eosinophil protein was inferred from our gene microarray study of mouse eosinophilopoiesis. Here, we detect 47 kDa intracellular and approximately 60 kDa secretory forms of PAI-2 in purified human eosinophil extracts. PAI-2 is present at variable concentrations in eosinophil lysates, ranging from 30 to 444 ng/10(6) cells, with a mean of 182 ng/10(6) cells from 10 normal donors, which is the highest per-cell concentration among all leukocyte subtypes evaluated. Enzymatic assay confirmed that eosinophil-derived PAI-2 is biologically active and inhibits activation of its preferred substrate, urokinase. Immunohistochemical and immunogold staining demonstrated PAI-2 localization in eosinophil-specific granules. Immunoreactive PAI-2 was detected in extracellular deposits in and around the eosinophil-enriched granuloma tissue encapsulating the parasitic egg in livers of wild-type mice infected with the helminthic parasite Schistosoma mansoni. Among the possibilities, we consider a role for eosinophil-derived PAI-2 in inflammation and remodeling associated with parasitic infection as well as allergic airways disease, respiratory virus infection, and host responses to tumors and metastasis in vivo.

  4. Immunomodulatory effects of selected Malaysian plants on the CD18/11a expression and phagocytosis activities of leukocytes

    Institute of Scientific and Technical Information of China (English)

    Nurul; Hikmah; Harun; Abdi; Wira; Septama; Ibrahim; Jantan

    2015-01-01

    Objective:To investigate the effects of 20 methanolic extracts from Malaysian selected plants on CD18/11 a expression and phagocytosis activity of leukocytes using flow cytometry analysis.Methods:The effects of methanolic extracts on CD18/11 a expression and phagocytosis of leukocytes were measured by labelling the cells with CD18-fluorescein isolhiocyanaie and ingestion labelled with Escherichia coli-fluorescein isothiocyanate and then analyzed using flow cytometer.Results:About 12 out of 20 methanolic extracts of selected Malaysian medicinal plants significantly(P≤0.05) inhibited the CD18/1 la expression of leukocytes at both concentrations of 6.25 μg/mL and 100 μg/mL in dose dependent manner.The most active inhibitory was shown in Citrus aurantifolia(Christm.) Swingle and Alpinia galangal(L.) Willd.at dosage 100ug/mL.Moreover,the Orthosiphon aristatus(Blume) Miq(O.aristatus).showed the highest stimulatory activity at the concentration of 100 μg/mL.Other than that,four plant extracts significantly(P<0.05) rose the phagocytosis activities of leukocytes in dose dependent manner.However,Annona muricata L.and O.aristatus showed the highest stimulated activities at the 100 pg/mL concentration.Conclusions:The results suggest that methanolic extracts of Cirrus aurantifolia.Alpinia gaiangal,O.aristatus and Annona muricata are able to modulate innate immune system and can potentially be recognized as therapeutic agents for modulating immune system.

  5. Antifungal activity against Cryptococcus neoformans strains and genotoxicity assessment in human leukocyte cells of Euphorbia tirucalli L.

    Directory of Open Access Journals (Sweden)

    Luís Flávio Souza de Oliveira

    2014-12-01

    Full Text Available In the last times, focus on plant research has increased all over the world. Euphorbia tirucalli L., a plant known popularly as Aveloz, and originally used in Africa, has been drawing attention for its use in the United States and Latin America, both for use as an ornamental plant and as a medicinal plant. E. tirucalli L. is a member of the family Euphorbiaceae and contains many diterpenoids and triterpenoids, in particular phorbol esters, apparently the main constituent of this plant, which are assumed to be responsible for their activities in vivo and in vitro. The in vitro antifungal activities of Euphorbia tirucalli (L. against opportunistic yeasts were studied using microbroth dilution assay. The results showed that aqueous extract and latex preparation were effective against ten clinical strains of Cryptococcus neoformans in vitro (Latex and extract MIC range of 3.2 - > 411 µg/mL. Aiming the safe use in humans, the genotoxic effects of E. tirucalli were evaluated in human leukocytes cells. Our data show that both aqueous extract and latex preparation have no genotoxic effect in human leukocytes cells in vitro. Although the results cannot be extrapolated by itself for use in vivo, they suggest a good perspective for a therapeutic application in future. In conclusion, our results show that the aqueous extract and latex preparation from E. tirucalli L. are antifungal agents effectives against several strains of C. neoformans and do not provoke DNA damage in human leukocyte cells, considering the concentrations tested.

  6. Allogeneic leukocytes in cardiac surgery: Good or bad?

    Directory of Open Access Journals (Sweden)

    Anneke Brand

    2011-09-01

    Full Text Available Worldwide, cardiac surgery is a common procedure requiring a large quantity of allogeneic blood products, which are associated with postoperative complications. Leukocytes present in blood products may play a role in these complications, which are referred to as transfusion-related immunomodulation (TRIM. Several randomized controlled trials (RCTs in different settings investigated the effects of allogeneic leukocytes in red blood cells (RBCs. Cardiac surgery studies reported a reduction in postoperative infections and mortality in patients that received leukocyte-reduced RBCs compared with leukocyte-containing RBCs; this was mainly due to more deaths due to infections and multiple organ dysfunction syndrome (MODS in the group that received leukocyte-containing RBCs. Patients with postoperative complications had higher concentrations of inflammatory mediators. These findings suggest that leukocyte-containing transfusion during cardiac surgery induces a second insult to the systemic inflammatory response. In the present review we discuss the possible role of blood transfusions in cardiac surgery. Especially, we focus on the possible role of allogeneic leukocytes associated with postoperative complications after cardiac surgery.

  7. Influence of Magnetite Nanoparticles on Human Leukocyte Activity

    Science.gov (United States)

    Džarová, Anežka; Dubničková, Martina; Závišová, Vlasta; Koneracká, Martina; Kopčanský, Peter; Gojzewski, Hubert; Timko, Milan

    2010-12-01

    Chemically synthesized magnetite particles coated by sodium oleate and PEG (MNP), and magnetosomes (MS) influence the process of phagocytosis and the metabolic activity (lysozyme and peroxidase activity) in leukocytes. Lysozyme activity is oxygen-independent liquidation mechanisms of engulfed microorganism, peroxidase activity is an oxygen-dependent mechanism. Both tested types of nanoparticles lysed leukocyte cells during incubation. MNP at concentrations of 10 and 20 μg/mL lysed almost all leukocytes and their cell viability was in the 14±0.05% range. On the other hand MS begin to influence leukocytes activity at the concentration of 1 μg/ml and this influence grows with increasing concentration up to 20 μg/ml. MS are more suitable for biological applications than MNP which are more aggressive material than MS. MS should not be used above 10 μg/mL.

  8. Lectins in extracts of certain Polygonaceae seed precipitate animal and human serums.

    Science.gov (United States)

    Hanan, E B; Spindler, J W

    1968-06-28

    Seeds of four species of Polygonaceae were tested for lectins that precipitate human and animal serums. Rumex crispus, Polygonum convolvulus, and Polygonum pennsylvanicum developed specific precipitate bands on double diffusion on agar gel plates. These bands were enhanced and increased in number when extracts were tested against serums from patients with certain diseases. When tested against lyophilized serum, no precipitate bands developed. The active substance cannot be dialyzed through cellulose membrane against running tap water for 16 hours, and it is heat stable. Extracts from Fagopyrum esculentum developed no precipitate bands.

  9. Iron, zinc and calcium dialyzability from extruded product based on whole grain amaranth (Amaranthus caudatus and Amaranthus cruentus) and amaranth/Zea mays blends.

    Science.gov (United States)

    Galan, María Gimena; Drago, Silvina Rosa; Armada, Margarita; José, Rolando González

    2013-06-01

    Amaranth is a Native American grain appreciated for its high nutritional properties including high mineral content. The aim of this study was to evaluate the availability of Fe, Zn and Ca from extruded products made with two varieties of amaranth and their mixtures with maize at two levels of replacement. Mineral availability was estimated using dialyzability method. The contents of Fe (64.0-84.0 mg/kg), Ca (1977.5-2348.8 mg/kg) and Zn (30.0-32.1 mg/kg) were higher in amaranth than in maize products (6.2, 19.1, 9.7 mg/kg, respectively). Mineral availability was in the range of (2.0-3.6%), (3.3-11.1%) and (1.6-11.4%) for Fe, Ca and Zn, respectively. Extruded amaranth and amaranth/maize products provide higher amount of Fe and Ca than extruded maize. Extruded amaranth products and amaranth addition to maize could be an interesting way to increase nutritional value of extruded products.

  10. C-type natriuretic peptide inhibits leukocyte recruitment and platelet-leukocyte interactions via suppression of P-selectin expression

    Science.gov (United States)

    Scotland, Ramona S.; Cohen, Marc; Foster, Paul; Lovell, Matthew; Mathur, Anthony; Ahluwalia, Amrita; Hobbs, Adrian J.

    2005-10-01

    The multifaceted process of immune cell recruitment to sites of tissue injury is key to the development of an inflammatory response and involved in the pathogenesis of numerous cardiovascular disorders. We recently identified C-type natriuretic peptide (CNP) as an important endothelium-derived mediator that regulates vascular tone and protects against myocardial ischemia/reperfusion injury. Herein, we investigated whether CNP inhibits leukocyte recruitment and platelet aggregation and thereby exerts a potential antiinflammatory influence on the blood vessel wall. We assessed the effects of CNP on leukocyte-endothelial cell interactions in mouse mesenteric postcapillary venules in vivo in animals with high basal leukocyte activation (endothelial nitric oxide synthase knockout mice, eNOS-/-) or under acute inflammatory conditions (induced by interleukin-1 or histamine). CNP suppressed basal leukocyte rolling in eNOS-/- mice in a rapid, reversible, and concentration-dependent manner. These effects of CNP were mimicked by the selective natriuretic peptide receptor-C agonist cANF4-23. CNP also suppressed leukocyte rolling induced by IL-1 or histamine, inhibited platelet-leukocyte interactions, and prevented thrombin-induced platelet aggregation of human blood. Furthermore, analysis of human umbilical vein endothelial cells, leukocytes, and platelets revealed that CNP selectively attenuates expression of P-selectin. Thus, CNP is a modulator of acute inflammation in the blood vessel wall characterized by leukocyte and platelet activation. These antiinflammatory effects appear to be mediated, at least in part, via suppression of P-selectin expression. These observations suggest that endothelial CNP might maintain an anti-atherogenic influence on the blood vessel wall and represent a target for therapeutic intervention in inflammatory cardiovascular disorders. endothelium | natriuretic peptide receptor type C | atherosclerosis | thrombosis

  11. Genomic signatures characterize leukocyte infiltration in myositis muscles

    Directory of Open Access Journals (Sweden)

    Zhu Wei

    2012-11-01

    Full Text Available Abstract Background Leukocyte infiltration plays an important role in the pathogenesis and progression of myositis, and is highly associated with disease severity. Currently, there is a lack of: efficacious therapies for myositis; understanding of the molecular features important for disease pathogenesis; and potential molecular biomarkers for characterizing inflammatory myopathies to aid in clinical development. Methods In this study, we developed a simple model and predicted that 1 leukocyte-specific transcripts (including both protein-coding transcripts and microRNAs should be coherently overexpressed in myositis muscle and 2 the level of over-expression of these transcripts should be correlated with leukocyte infiltration. We applied this model to assess immune cell infiltration in myositis by examining mRNA and microRNA (miRNA expression profiles in muscle biopsies from 31 myositis patients and 5 normal controls. Results Several gene signatures, including a leukocyte index, type 1 interferon (IFN, MHC class I, and immunoglobulin signature, were developed to characterize myositis patients at the molecular level. The leukocyte index, consisting of genes predominantly associated with immune function, displayed strong concordance with pathological assessment of immune cell infiltration. This leukocyte index was subsequently utilized to differentiate transcriptional changes due to leukocyte infiltration from other alterations in myositis muscle. Results from this differentiation revealed biologically relevant differences in the relationship between the type 1 IFN pathway, miR-146a, and leukocyte infiltration within various myositis subtypes. Conclusions Results indicate that a likely interaction between miR-146a expression and the type 1 IFN pathway is confounded by the level of leukocyte infiltration into muscle tissue. Although the role of miR-146a in myositis remains uncertain, our results highlight the potential benefit of deconvoluting the

  12. Expression of β2-integrin on leukocytes in liver cirrhosis

    Institute of Scientific and Technical Information of China (English)

    Anatol Panasiuk; Janusz Zak; Elzbieta Maciorkowska; Bozena Panasiuk; Danuta Prokopowicz

    2006-01-01

    AIM: To analyze β2-integrin expression on blood leukocytes in liver cirrhosis.METHODS: In 40 patients with liver cirrhosis and 20healthy individuals, the evaluation of expression of CD11a (LFA-1α), CD11b (Mac-1α), CD11c (αX) and CD49d (VLA-4α) on peripheral blood leukocytes was performed using flow cytometry. The analysis was carried out in groups of patients divided into B and C according to Child-Pugh's classification.RESULTS: An increased CD11a, CD11b, CD11c and CD49d integrin expression was observed on peripheral blood leukocytes in liver cirrhosis. The integrin levels were elevated as the advancement of liver failure progressed. The highest expression of integrins occurred predominantly on monocytes. A slight expression of VLA-4 was found on lymphocytes and granulocytes and it increased together with liver failure. A positive correlation was noted between median intensity of fluorescence (MIF) expression on polymorphonuclear cells of CD11a and CD11c and CD49d (r = 0.42, P < 0.01; r = 053, P < 0.01, respectively) in liver cirrhosis stage C. However,no correlation was observed between integrin expression on leukocytes. The concentrations of sICAM-1, sVCAM-1,and TNFα, were significantly elevated in liver cirrhosis.CONCLUSION: β2-integrin expression on leukocytes increases in liver cirrhosis decompensated as the stage of liver failure increases, which is a result of permanent activation of leukocytes circulating through the inflamed liver environment. β2-integrin expression on circulating leukocytes can intensify liver cirrhosis.

  13. Attempted Depletion of Passenger Leukocytes by Irradiation in Pigs

    Directory of Open Access Journals (Sweden)

    Hao-Chih Tai

    2011-01-01

    Full Text Available Allograft/xenograft rejection is associated with “passenger leukocyte” migration from the organ into recipient lymph nodes. In Study 1, we attempted to deplete leukocytes from potential kidney “donor” pigs, using two regimens of total body irradiation. A dose of 700 cGy was administered, followed by either 800 cGy (“low-dose” or 1,300 cGy (“high dose” with the kidneys shielded. Neither regimen was entirely successful in depleting all leukocytes, although remaining T and 8 cell numbers were negligible. Study 2 was aimed at providing an indication of whether near-complete depletion of leukocytes had any major impact on kidney allograft survival. In non-immunosuppressed recipient pigs, survival of a kidney from a donor that received high-dose irradiation was compared with that of a kidney taken from a non-irradiated donor. Kidney graft survival was 9 and 7 days, respectively, suggesting that depletion had little impact on graft survival. The lack of effect may have been related to (i inadequate depletion of passenger leukocytes, thus not preventing a direct T cell response, (ii the presence of dead or dying leukocytes (antigens, thus not preventing an indirect T cell response, or (iii constitutive expression of MHC class II and B7 molecules on the porcine vascular endothelium, activating recipient T cells.

  14. Contour Detection of Leukocyte Cell Nucleus Using Morphological Image

    Science.gov (United States)

    Supriyanti, R.; Satrio, G. P.; Ramadhani, Y.; Siswandari, W.

    2017-04-01

    Leukocytes are blood cells that do not contain color pigments. Leukocyte function to the tool body’s defenses. Abnormal forms of leukocytes can be a sign of serious diseases such example is leukemia. Most laboratories still use cell morphology examination to assist the diagnosis of illness associated with white blood cells such example is leukemia because of limited resources, both infrastructure, and human resources as happens in developing nations, such as Indonesia. This examination is less expensive and quicker process. However, morphological review requires the expertise of a specialist clinical pathology were limited. This process is sometimes less valid cause in some cases trying to differentiate morphology blast cells into the type of myoblasts, lymphoblast, monoblast, or erythroblast thus potentially misdiagnosis. The goal of this research is to develop a detection device types of blood cells automatically as lower-priced, easy to use and accurate so that the tool can be distributed across all units in existing health services throughout Indonesia and in particular for remote areas. However, because the variables used in the identification of abnormal leukocytes are very complex, in this paper, we emphasize on the contour detection of leukocyte cell nucleus using the morphological image. The results show that this method is promising for further development.

  15. Crossing the Vascular Wall: Common and Unique Mechanisms Exploited by Different Leukocyte Subsets during Extravasation

    Directory of Open Access Journals (Sweden)

    Michael Schnoor

    2015-01-01

    Full Text Available Leukocyte extravasation is one of the essential and first steps during the initiation of inflammation. Therefore, a better understanding of the key molecules that regulate this process may help to develop novel therapeutics for treatment of inflammation-based diseases such as atherosclerosis or rheumatoid arthritis. The endothelial adhesion molecules ICAM-1 and VCAM-1 are known as the central mediators of leukocyte adhesion to and transmigration across the endothelium. Engagement of these molecules by their leukocyte integrin receptors initiates the activation of several signaling pathways within both leukocytes and endothelium. Several of such events have been described to occur during transendothelial migration of all leukocyte subsets, whereas other mechanisms are known only for a single leukocyte subset. Here, we summarize current knowledge on regulatory mechanisms of leukocyte extravasation from a leukocyte and endothelial point of view, respectively. Specifically, we will focus on highlighting common and unique mechanisms that specific leukocyte subsets exploit to succeed in crossing endothelial monolayers.

  16. Improving diagnosis of appendicitis. Early autologous leukocyte scanning.

    Science.gov (United States)

    DeLaney, A R; Raviola, C A; Weber, P N; McDonald, P T; Navarro, D A; Jasko, I

    1989-10-01

    A prospective nonrandomized study investigating the accuracy and utility of autologous leukocyte scanning in the diagnosis of apendicitis was performed. One hundred patients in whom the clinical diagnosis of appendicitis was uncertain underwent indium 111 oxyquinoline labelling of autologous leukocytes and underwent scanning 2 hours following reinjection. Of 32 patients with proved appendicitis, three scans revealed normal results (false-negative rate, 0.09). Of 68 patients without appendicitis, three scans had positive results (false-positive rate, 0.03; sensitivity, 0.91; specificity, 0.97; predictive value of positive scan, 0.94; predictive value of negative scan, 0.96; and overall accuracy, 0.95). Scan results altered clinical decisions in 19 patients. In 13 cases, the scan produced images consistent with diagnoses other than appendicitis, expediting appropriate management. Early-imaging111 In oxyquinoline autologous leukocyte scanning is a practical and highly accurate adjunct for diagnosing appendicitis.

  17. Leukocyte depletion during CPB: effects on inflammation and lung function.

    Science.gov (United States)

    de Amorim, Célio Gomes; Malbouisson, Luiz Marcelo Sá; da Silva, Francisco Costa; Fiorelli, Alfredo Inácio; Murakami, Caroline Kameio Fernandes; Carmona, Maria José Carvalho

    2014-02-01

    Cardiopulmonary bypass (CPB) is related to inflammatory response and pulmonary dysfunction. The aim of this study was to evaluate the effects of CPB leukocyte filtration on inflammation and lung function after coronary artery bypass grafting (CABG). A prospective randomized study was performed to compare CABG patients undergoing CPB leukocyte filtration (n = 9) or standard CPB (n = 11). Computed tomography, oxygenation, leukocyte count, hemodynamic data, PaO2/FiO2, shunt fraction, interleukins, elastase, and myeloperoxidase were evaluated. Data were analyzed using two-factor ANOVA for repeated measurements. The filtered group showed lower neutrophil counts up to 50 min of CPB, lower shunt fraction up to 6 h after surgery, and lower levels of IL-10 at the end of surgery (p CPB results in neutrophil sequestration by a short time, decreased IL-10 serum levels, and lower worsening of lung function only temporarily.

  18. Platelet subpopulation bearing leukocyte specific antigen and tissue factor.

    Science.gov (United States)

    Gabbasov, Z A; Saburova, O S; Antonova, O A; Golubeva, N V; Khaspekova, S G; Shustova, O N; Zyuryaev, I T; Ruda, M Ya; Mazurov, A V

    2016-11-01

    Platelets bearing leukocyte antigen CD45 were identified in the blood of patients with myocardial infarction (MI) and healthy donors by flow cytofluorimetry. Part of these platelets contained tissue factor (TF)-primary initiator of blood clotting. The number of CD45(+) and CD45(+)/TF(+) platelets in MI patients at the first day was comparable with their level in healthy donors, but was increased at 8-12 days after MI onset. At that time in some patients the amount of CD45(+) and CD45(+)/TF(+) platelets reached 5-6 and 2-3% of their total number. It is assumed that CD45(+)/TF(+) platelets could be formed as a result of platelet interaction with leukocytes or leukocyte produced membrane microparticles.

  19. In Vivo Quantitation of Local Anesthetic Suppression of Leukocyte Adherence

    Science.gov (United States)

    Giddon, D. B.; Lindhe, J.

    1972-01-01

    Using intravital microscopy, topically applied amide-type local anesthetics suppressed the adherence of leukocytes to the venular endothelium within surgical defects in the hamster cheek pouch. The response was reversible with physiologic saline and was localized to venules within the defect. Quantitation in terms of the percent of initially adhering leukocytes remaining in place on the venule wall at each minute following application of lidocaine and physiologic saline, respectively, revealed the suppression to be reliably related to the concentration, viz: 20.0 >10.0 >5.0 >0.0 mg ml of commercially available Xylocaine® (lidocaine) HCl. ImagesFig 1Fig 1 PMID:5049429

  20. Inhibitory Effect of Helicteres gardneriana Ethanol Extract on Acute Inflammation

    Directory of Open Access Journals (Sweden)

    Juliana Oliveira de Melo

    2012-01-01

    Full Text Available The anti-inflammatory effect of an ethanol extract of Helicteres gardneriana (Nees Castiglioni was assayed in experimental models of pleurisy and microcirculation in situ. Treatment of animals with 500 mg/kg body weight reduced the exudate volume (35% reduction induced by intrapleural injection of carrageenan and the migration of polymorphonuclear cells into the inflamed pleural cavity of rats (40%. Additionally, rolling and adhesion of leukocytes and the number of leukocytes that migrated toward the perivascular space in response to the carrageenan injection were decreased by the extract (500 mg/kg. These data demonstrate the anti-inflammatory effect of the ethanol extract of Helicteres gardneriana and imply that inhibition of leukocyte-endothelial interactions is important in the extract's mechanism of action.

  1. The Effect of Hemiscorpius lepturus (Scorpionida: Hemiscorpiidae Venom on Leukocytes and the Leukocyte Subgroups in Peripheral Blood of Rat

    Directory of Open Access Journals (Sweden)

    Mehri Ghafourian

    2016-01-01

    Full Text Available Background: The aim of this study was to investigate the effect of Hemiscorpius lepturus venom on leukocytes and the leukocyte subgroups in peripheral blood of rat.Methods: In this experimental study, sixty N-Mari rats were divided into three groups of 20 rats. Then the rats in each group were divided into four subgroups based on the blood sampling time that was 2, 6, 24 and 48 hours after the venom injection, respectively. The control group did not receive anything, however, the first and the second ex­perimental groups received 0.1 and 0.01mg/kg of venom, subcutaneously. In accordance with a designated four sam­pling times, the blood sampling was carried out in three groups. After RBC lysis, the leukocytes and leukocyte sub­populations were determined and counted using appropriate hematological standard methods.Results: The leukocyte and the neutrophil count at two (P<0.05, six (P<0.01 and 24 (P<0.05 hours after the venom injection showed a significant decline compared with the control group, this decrease was significant at the dose of 0.1 mg/kg until 48 hours after the venom injection (P<0.05. The lymphocyte count showed a significant decline throughout the all hours of the experiment, compared with the control group (P<0.05.Conclusion: Leukocytes are probably affected by the cytotoxicity effect of the H. lepturus venom in a dose-dependent manner. This could be a wakeup call for the medical staff to perform quick and accurate treatment in the least time possible.

  2. Production and characterization of monoclonal antibodies against mink leukocytes

    DEFF Research Database (Denmark)

    Chen, W.S.; Pedersen, Mikael; Gram-Nielsen, S.

    1997-01-01

    Three monoclonal antibodies (mAbs) were generated against mink leukocytes. One antibody reacted with all T lymphocytes, one with all monocytes and one had platelet reactivity. Under reducing conditions, the T lymphocyte reactive antibody immunoprecipitated 18 kDa, 23 kDa, 25 kDa and 32-40 kDa pol...

  3. Improved survival of newborns receiving leukocyte transfusions for sepsis

    Energy Technology Data Exchange (ETDEWEB)

    Cairo, M.S.; Rucker, R.; Bennetts, G.A.; Hicks, D.; Worcester, C.; Amlie, R.; Johnson, S.; Katz, J.

    1984-11-01

    To determine the role of polymorphonuclear (PMN) leukocyte transfusions in neonates with sepsis, 23 consecutive newborns were prospectively randomly selected during an 18-month period in a treatment plan to receive polymorphonuclear leukocyte transfusions with supportive care or supportive care alone. Thirteen neonates received transfusions every 12 hours for a total of five transfusions. Each transfusion consisting of 15 mL/kg of polymorphonuclear leukocytes was subjected to 1,500 rads of radiation. The polymorphonuclear leukocytes were obtained by continuous-flow centrifugation leukapheresis and contained 0.5 to 1.0 X 10(9) granulocytes per 15 mL with less than 10% lymphocytes. Positive findings on blood cultures were obtained in 14/23 patients and seven were randomly selected for each treatment group. Absolute granulocyte counts were less than 1,500/microL in 13 patients but tibial bone marrow examinations revealed that the neutrophil supply pool was depleted in only three patients. The survival was significantly greater in the treatment group compared with the group that did not receive transfusions.

  4. Activation of peripheral leukocytes in rat pregnancy and experimental preeclampsia

    NARCIS (Netherlands)

    Faas, MM; Schuiling, GA; Linton, EA; Sargent, IL; Redman, CWG

    OBJECTIVE: The aim of this study was to search for activation markers of peripheral leukocytes in experimental preeclampsia in the rat. STUDY DESIGN: Experimental preeclampsia was induced in 14-day-pregnant rats by infusion of endotoxin (1.0 mu g/kg body weight). For comparison, rats with normal

  5. Upregulation of Leukocytic Syncytin-1 in Acute Myeloid Leukemia Patients.

    Science.gov (United States)

    Sun, Yi; Zhu, Hongyan; Song, Jianxin; Jiang, Yaxian; Ouyang, Hongmei; Huang, Rongzhong; Zhang, Guiqian; Fan, Xin; Tao, Rui; Jiang, Jie; Niu, Hua

    2016-07-09

    BACKGROUND Syncytin-1, a cell membrane-localizing fusogen, is abnormally expressed in several cancers, including endometrial cancer, breast cancer, and leukemia. Although abnormal syncytin-1 expression has been detected in two-thirds of leukemia blood samples, its expression profile in acute leukemia patients has not yet been analyzed. MATERIAL AND METHODS Bone marrow samples from 50 acute myelogenous leukemia (AML) cases and 14 B-cell acute lymphocytic leukemia (B-cell ALL) patients were subjected to flow cytometry to assess leukocyte type distributions and leukocytic syncytin-1 surface expression. RT-PCR was applied to assess leukocytic syncytin-1 mRNA expression. Statistical analysis was applied to compare syncytin-1 expression between AML and B-cell ALL patients across blasts, granulocytes, lymphocytes, and monocytes as well as to determine clinical factors statistically associated with changes in syncytin-1 expression. RESULTS The leukocyte type distributions of the AML and B-cell ALL cohorts highly overlapped, with an observable difference in blast distribution between the 2 cohorts. The AML cohort displayed significantly greater syncytin-1 surface and mRNA expression (pphenotype and the acute monocytic leukemia phenotype in particular.

  6. Differential MSC activation leads to distinct mononuclear leukocyte binding mechanisms

    Science.gov (United States)

    Kota, Daniel J.; Dicarlo, Bryan; Hetz, Robert A.; Smith, Philippa; Cox, Charles S.; Olson, Scott D.

    2014-04-01

    Advances in the field of Multipotent Mesenchymal Stromal cell (MSC) biology have demonstrated that MSCs can improve disease outcome when `activated' to exert immunomodulatory effects. However, the precise mechanisms modulating MSC-immune cells interactions remain largely elusive. In here, we activated MSC based on a recent polarization paradigm, in which MSCs can be polarized towards a pro- or anti-inflammatory phenotype depending on the Toll-like receptor stimulated, to dissect the mechanisms through which MSCs physically interact with and modulate leukocytes in this context. Our data show that MSCs activated through the Toll-like receptor (TLR) 4 pathway increased VCAM-1 and ICAM-1 dependent binding of leukocytes. On the other hand, TLR3 stimulation strongly increases leukocytes affinity to MSC comparatively, through the formation of cable-like hyaluronic acid structures. In addition, TLR4 activation elicited secretion of pro-inflammatory mediators by MSCs, whereas TLR3-activated MSCs displayed a milder pro-inflammatory phenotype, similar to inactivated MSCs. However, the differently activated MSCs maintained their ability to suppress leukocyte activation at similar levels in our in vitro model, and this immunomodulatory property was shown here to be partially mediated by prostaglandin. These results reinforce the concept that alternate activation profiles control MSC responses and may impact the therapeutic use of MSCs.

  7. Radiolabelled leukocytes for imaging inflammation: how radiochemistry affects clinical use.

    Science.gov (United States)

    Ballinger, J R; Gnanasegaran, G

    2005-12-01

    Indium-111((111)In)-labelled leukocytes were introduced for imaging inflammation about 25 years ago. A few years later methods to label leukocytes with Technetium-99m ((99m)Tc) were developed, but the two radiolabels cannot be used interchangeably. The amount of radioactivity which can be administered with (111)In is low, because of its 67-h half-life and associated radiation dose. This results in low count density in images. However, (111)In labelling is very stable, with binding to intracellular macromolecules and particulates, and there is minimal urinary or faecal excretion. In contrast, (99m)Tc has a half-life of 6 h and can be administered in higher doses, resulting in improved image quality. However, (99m)Tc labelling is less stable because the trapped form is soluble and there is excretion of (99m)Tc through both the kidneys and intestine, which limits imaging of disease in the abdomen except at early times. There is interest in extending inflammation imaging to PET. Although leukocytes can be labelled with (18)F-FDG, its half-life and stability are not optimal and radiometals such as Copper-64 are being evaluated. Despite the laborious nature of leukocyte labelling, it has yet to be replaced by direct injection agents.

  8. Leukocyte telomere length dynamics in women and men

    DEFF Research Database (Denmark)

    Dalgård, Christine; Benetos, Athanase; Verhulst, Simon;

    2015-01-01

    BACKGROUND: A longer leukocyte telomere length (LTL) in women than men has been attributed to a slow rate of LTL attrition in women, perhaps due to high estrogen exposure during the premenopausal period. METHODS: To test this premise we performed a longitudinal study (an average follow-up of 12...

  9. Longitudinal Changes in Leukocyte Telomere Length and Mortality in Humans

    DEFF Research Database (Denmark)

    Bendix, Laila; Thinggaard, Mikael; Fenger, Mogens;

    2014-01-01

    Leukocyte telomere length (LTL) ostensibly shortens with age and has been moderately associated with mortality. In humans, these findings have come almost solely from cross-sectional studies. Only recently has LTL shortening within individuals been analyzed in longitudinal studies. Such studies...... are relevant to establish LTL dynamics as biomarkers of mortality as well as to disentangle the causality of telomeres on aging....

  10. VARIABILITY OF LEUKOCYTE COUNT AS AN INFLAMMATORY RESPONSE AMONG HYPERTENSIVES

    Directory of Open Access Journals (Sweden)

    Brinda

    2015-12-01

    Full Text Available INTRODUCTION Hypertension, a globally prevailing non communicable disease has been linked to various pathophysiological mechanisms. The new spotlight is on the inflammatory mechanism, which has been embarked on this study by utilising the White blood cell counts as the predominant marker. The pathogenesis behind this concept is the endothelial alteration which occurs in hypertension, causing increased oxidative stress and release of inflammatory mediators and further causing damage to the vascular walls and may result in sustained hypertension. OBJECTIVE To substantiate the difference in leukocyte count among the hypertensives and normotensives, to assess the variation in the leukocyte count among varied grades of hypertension. METHODS 80 subjects and 40 controls of both sex and age between 20-60 years were taken. Subjects with other chronic medical disorders, pregnancy, menstruation phase, history of infections in past 3 months, obesity, smokers and regular alcoholics were excluded. Three recordings of blood pressure were measured with a standard mercury sphygmomanometer. The differential leukocyte count was estimated with an automated counter. RESULTS Statistical analysis was done using independent sample t-test and one-way ANOVA. Total Leukocyte count (Mean = 8397.50 ± 1666.57 was raised significantly among the hypertensives, with P value <0.001. Comparison was done on the total count with the varied hypertensive years, which was suggestive of increase in total count among the pre hypertensives (systolic pressure and there was linear increase in total count with diastolic pressure though not statistically significant. CONCLUSION The elevation of white blood cell count is suggestive of underlying inflammatory mechanism among the hypertensives. The onset of inflammation could be at the onset of hypertension, as there is rise in total leukocyte count among the pre hypertensives which could also lead to sustained hypertension.

  11. Efficiency and safety of leukocyte filtration during cardiopulmonary bypass for cardiac surgery

    NARCIS (Netherlands)

    Smit, JJJ; de Vries, AJ; Gu, YJ; van Oeveren, W

    1999-01-01

    Background. Leukocyte filtration of systemic blood during cardiopulmonary bypass surgery to reduce post-operative morbidity has not yet been established because of the enormous leukocyte release from the third space. This study was designed to examine the efficiency and safety of leukocyte filtratio

  12. File list: InP.Bld.20.AllAg.Polymorphonuclear_leukocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available InP.Bld.20.AllAg.Polymorphonuclear_leukocytes hg19 Input control Blood Polymorphonu...clear leukocytes http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/InP.Bld.20.AllAg.Polymorphonuclear_leukocytes.bed ...

  13. File list: DNS.Bld.10.AllAg.Polymorphonuclear_leukocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available DNS.Bld.10.AllAg.Polymorphonuclear_leukocytes hg19 DNase-seq Blood Polymorphonuclea...r leukocytes http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/DNS.Bld.10.AllAg.Polymorphonuclear_leukocytes.bed ...

  14. File list: Oth.Bld.50.AllAg.Polymorphonuclear_leukocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Oth.Bld.50.AllAg.Polymorphonuclear_leukocytes hg19 TFs and others Blood Polymorphonu...clear leukocytes http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/Oth.Bld.50.AllAg.Polymorphonuclear_leukocytes.bed ...

  15. File list: NoD.Bld.20.AllAg.Polymorphonuclear_leukocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available NoD.Bld.20.AllAg.Polymorphonuclear_leukocytes hg19 No description Blood Polymorphonu...clear leukocytes SRX1016682,SRX1016679,SRX1016680,SRX1016681 http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/NoD.Bld.20.AllAg.Polymorphonuclear_leukocytes.bed ...

  16. File list: ALL.Bld.05.AllAg.Polymorphonuclear_leukocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available ALL.Bld.05.AllAg.Polymorphonuclear_leukocytes hg19 All antigens Blood Polymorphonuc...lear leukocytes SRX1016679,SRX1016682,SRX1016681,SRX1016680 http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/ALL.Bld.05.AllAg.Polymorphonuclear_leukocytes.bed ...

  17. File list: ALL.Bld.50.AllAg.Polymorphonuclear_leukocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available ALL.Bld.50.AllAg.Polymorphonuclear_leukocytes hg19 All antigens Blood Polymorphonuc...lear leukocytes SRX1016682,SRX1016679,SRX1016680,SRX1016681 http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/ALL.Bld.50.AllAg.Polymorphonuclear_leukocytes.bed ...

  18. File list: Oth.Bld.05.AllAg.Polymorphonuclear_leukocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Oth.Bld.05.AllAg.Polymorphonuclear_leukocytes hg19 TFs and others Blood Polymorphonu...clear leukocytes http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/Oth.Bld.05.AllAg.Polymorphonuclear_leukocytes.bed ...

  19. File list: DNS.Bld.05.AllAg.Polymorphonuclear_leukocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available DNS.Bld.05.AllAg.Polymorphonuclear_leukocytes hg19 DNase-seq Blood Polymorphonuclea...r leukocytes http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/DNS.Bld.05.AllAg.Polymorphonuclear_leukocytes.bed ...

  20. File list: NoD.Bld.05.AllAg.Polymorphonuclear_leukocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available NoD.Bld.05.AllAg.Polymorphonuclear_leukocytes hg19 No description Blood Polymorphonu...clear leukocytes SRX1016679,SRX1016682,SRX1016681,SRX1016680 http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/NoD.Bld.05.AllAg.Polymorphonuclear_leukocytes.bed ...

  1. File list: Oth.Bld.20.AllAg.Polymorphonuclear_leukocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Oth.Bld.20.AllAg.Polymorphonuclear_leukocytes hg19 TFs and others Blood Polymorphonu...clear leukocytes http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/Oth.Bld.20.AllAg.Polymorphonuclear_leukocytes.bed ...

  2. File list: InP.Bld.10.AllAg.Polymorphonuclear_leukocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available InP.Bld.10.AllAg.Polymorphonuclear_leukocytes hg19 Input control Blood Polymorphonu...clear leukocytes http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/InP.Bld.10.AllAg.Polymorphonuclear_leukocytes.bed ...

  3. File list: NoD.Bld.50.AllAg.Polymorphonuclear_leukocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available NoD.Bld.50.AllAg.Polymorphonuclear_leukocytes hg19 No description Blood Polymorphonu...clear leukocytes SRX1016682,SRX1016679,SRX1016680,SRX1016681 http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/NoD.Bld.50.AllAg.Polymorphonuclear_leukocytes.bed ...

  4. File list: Pol.Bld.20.AllAg.Polymorphonuclear_leukocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Pol.Bld.20.AllAg.Polymorphonuclear_leukocytes hg19 RNA polymerase Blood Polymorphonu...clear leukocytes http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/Pol.Bld.20.AllAg.Polymorphonuclear_leukocytes.bed ...

  5. File list: InP.Bld.50.AllAg.Polymorphonuclear_leukocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available InP.Bld.50.AllAg.Polymorphonuclear_leukocytes hg19 Input control Blood Polymorphonu...clear leukocytes http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/InP.Bld.50.AllAg.Polymorphonuclear_leukocytes.bed ...

  6. File list: ALL.Bld.20.AllAg.Polymorphonuclear_leukocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available ALL.Bld.20.AllAg.Polymorphonuclear_leukocytes hg19 All antigens Blood Polymorphonuc...lear leukocytes SRX1016682,SRX1016679,SRX1016680,SRX1016681 http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/ALL.Bld.20.AllAg.Polymorphonuclear_leukocytes.bed ...

  7. File list: ALL.Bld.10.AllAg.Polymorphonuclear_leukocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available ALL.Bld.10.AllAg.Polymorphonuclear_leukocytes hg19 All antigens Blood Polymorphonuc...lear leukocytes SRX1016682,SRX1016679,SRX1016681,SRX1016680 http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/ALL.Bld.10.AllAg.Polymorphonuclear_leukocytes.bed ...

  8. File list: DNS.Bld.20.AllAg.Polymorphonuclear_leukocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available DNS.Bld.20.AllAg.Polymorphonuclear_leukocytes hg19 DNase-seq Blood Polymorphonuclea...r leukocytes http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/DNS.Bld.20.AllAg.Polymorphonuclear_leukocytes.bed ...

  9. File list: DNS.Bld.50.AllAg.Polymorphonuclear_leukocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available DNS.Bld.50.AllAg.Polymorphonuclear_leukocytes hg19 DNase-seq Blood Polymorphonuclea...r leukocytes http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/DNS.Bld.50.AllAg.Polymorphonuclear_leukocytes.bed ...

  10. File list: Pol.Bld.50.AllAg.Polymorphonuclear_leukocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Pol.Bld.50.AllAg.Polymorphonuclear_leukocytes hg19 RNA polymerase Blood Polymorphonu...clear leukocytes http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/Pol.Bld.50.AllAg.Polymorphonuclear_leukocytes.bed ...

  11. File list: Pol.Bld.05.AllAg.Polymorphonuclear_leukocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Pol.Bld.05.AllAg.Polymorphonuclear_leukocytes hg19 RNA polymerase Blood Polymorphonu...clear leukocytes http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/Pol.Bld.05.AllAg.Polymorphonuclear_leukocytes.bed ...

  12. File list: InP.Bld.05.AllAg.Polymorphonuclear_leukocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available InP.Bld.05.AllAg.Polymorphonuclear_leukocytes hg19 Input control Blood Polymorphonu...clear leukocytes http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/InP.Bld.05.AllAg.Polymorphonuclear_leukocytes.bed ...

  13. File list: NoD.Bld.10.AllAg.Polymorphonuclear_leukocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available NoD.Bld.10.AllAg.Polymorphonuclear_leukocytes hg19 No description Blood Polymorphonu...clear leukocytes SRX1016682,SRX1016679,SRX1016681,SRX1016680 http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/NoD.Bld.10.AllAg.Polymorphonuclear_leukocytes.bed ...

  14. Age-related changes following in vitro stimulation with Rhodococcus equi of peripheral blood leukocytes from neonatal foals.

    Science.gov (United States)

    Kachroo, Priyanka; Ivanov, Ivan; Seabury, Ashley G; Liu, Mei; Chowdhary, Bhanu P; Cohen, Noah D

    2013-01-01

    Rhodococcus equi is an intracellular bacterium primarily known as an equine pathogen that infects young foals causing a pyogranulomatuous pneumonia. The molecular mechanisms mediating the immune response of foals to R. equi are not fully elucidated. Hence, global genomic high-throughput tools like gene expression microarrays might identify age-related gene expression signatures and molecular pathways that contribute to the immune mechanisms underlying the inherent susceptibility of foals to disease caused by R. equi. The objectives of this study were 2-fold: 1) to compare the expression profiles at specific ages of blood leukocytes from foals stimulated with virulent R. equi with those of unstimulated leukocytes; and, 2) to characterize the age-related changes in the gene expression profile associated with blood leukocytes in response to stimulation with virulent R. equi. Peripheral blood leukocytes were obtained from 6 foals within 24 hours (h) of birth (day 1) and 2, 4, and 8 weeks after birth. The samples were split, such that half were stimulated with live virulent R. equi, and the other half served as unstimulated control. RNA was extracted and the generated cDNA was labeled with fluorescent dyes for microarray hybridizations using an equine microarray. Our findings suggest that there is age-related differential expression of genes involved in host immune response and immunity. We found induction of genes critical for host immunity against pathogens (MHC class II) only at the later time-points (compared to birth). While it appears that foals up to 8-weeks of age are able to initiate a protective inflammatory response against the bacteria, relatively decreased expression of various other immune-related genes points toward inherent diminished immune responses closer to birth. These genes and pathways may contribute to disease susceptibility in foals if infected early in life, and might thus be targeted for developing preventative or therapeutic strategies.

  15. Age-related changes following in vitro stimulation with Rhodococcus equi of peripheral blood leukocytes from neonatal foals.

    Directory of Open Access Journals (Sweden)

    Priyanka Kachroo

    Full Text Available Rhodococcus equi is an intracellular bacterium primarily known as an equine pathogen that infects young foals causing a pyogranulomatuous pneumonia. The molecular mechanisms mediating the immune response of foals to R. equi are not fully elucidated. Hence, global genomic high-throughput tools like gene expression microarrays might identify age-related gene expression signatures and molecular pathways that contribute to the immune mechanisms underlying the inherent susceptibility of foals to disease caused by R. equi. The objectives of this study were 2-fold: 1 to compare the expression profiles at specific ages of blood leukocytes from foals stimulated with virulent R. equi with those of unstimulated leukocytes; and, 2 to characterize the age-related changes in the gene expression profile associated with blood leukocytes in response to stimulation with virulent R. equi. Peripheral blood leukocytes were obtained from 6 foals within 24 hours (h of birth (day 1 and 2, 4, and 8 weeks after birth. The samples were split, such that half were stimulated with live virulent R. equi, and the other half served as unstimulated control. RNA was extracted and the generated cDNA was labeled with fluorescent dyes for microarray hybridizations using an equine microarray. Our findings suggest that there is age-related differential expression of genes involved in host immune response and immunity. We found induction of genes critical for host immunity against pathogens (MHC class II only at the later time-points (compared to birth. While it appears that foals up to 8-weeks of age are able to initiate a protective inflammatory response against the bacteria, relatively decreased expression of various other immune-related genes points toward inherent diminished immune responses closer to birth. These genes and pathways may contribute to disease susceptibility in foals if infected early in life, and might thus be targeted for developing preventative or therapeutic

  16. Preparation of /sup 111/In leukocytes after hemolytic removal of erythrocytes

    Energy Technology Data Exchange (ETDEWEB)

    Karesh, S.M.; Henkin, R.E.

    1987-01-01

    An optimized procedure is described for isolation and high-efficiency radiolabeling of leukocytes using /sup 111/In-oxine. The chief advantages over conventional methods include virtually no loss of leukocytes during washing and separation steps; a significant reduction in the time required to prepare leukocytes for radiolabeling compared to non-hemolytic preparations; a 28% increase in the average labeling efficiency obtained using /sup 111/In-oxine; > 95% cell viability as measured by the trypan blue exclusion test; elimination of contaminating red blood cells from the leukocyte pellet prior to labeling; and 80% survivability at 15 min post injection (measured as per cent of blood activity on leukocyte fraction).

  17. Combined bone scintigraphy and indium-111 leukocyte scans in neuropathic foot disease

    Energy Technology Data Exchange (ETDEWEB)

    Schauwecker, D.S.; Park, H.M.; Burt, R.W.; Mock, B.H.; Wellman, H.N.

    1988-10-01

    It is difficult to diagnose osteomyelitis in the presence of neurotrophic osteoarthropathy. We performed combined (99mTc)MDP bone scans and indium-111 (111In) leukocyte studies on 35 patients who had radiographic evidence of neuropathic foot disease and clinically suspected osteomyelitis. The (111In)leukocyte study determined if there was an infection and the bone scan provided the anatomic landmarks so that the infection could be localized to the bone or the adjacent soft tissue. Seventeen patients had osteomyelitis and all showed increased (111In)leukocyte activity localized to the bone, giving a sensitivity of 100%. Among the 18 patients without osteomyelitis, eight had no accumulation of (111In)leukocytes, seven had the (111In)leukocyte activity correctly localized to the soft tissue, two had (111In)leukocyte activity mistakenly attributed to the bone, and one had (111In)leukocyte accumulation in a proven neuroma which was mistakenly attributed to bone. These three false-positive results for osteomyelitis reduced the specificity to 83%. Considering only the 27 patients with a positive (111In)leukocyte study, the combined bone scan and (111In)leukocyte study correctly localized the infection to the soft tissues or bone in 89%. Uninfected neurotrophic osteoarthropathy does not accumulate (111In)leukocytes. We found the combined bone scan and (111In) leukocyte study useful for the detection and localization of infection to soft tissue or bone in patients with neuropathic foot disease.

  18. Effect of radiographic contrast agents on leukocyte metabolic response

    Energy Technology Data Exchange (ETDEWEB)

    Hernanz-Schulman, M. [Dept. of Pediatric Radiology, Vanderbilt Children' s Hospital, Nashville, TN (United States); Vanholder, R.; Waterloos, M.A. [Dept. of Internal Medicine, Nephrology Section, University Hospital, Gent (Belgium); Hakim, R.; Schulman, G. [Department of Nephrology, Vanderbilt University Medical Center, Nashville, TN (United States)

    2000-06-01

    Barium, at clinical dilutions, causes a significant increase of baseline ''resting state'' phagocytic activity, which in turn leads to significant blunting of subsequent response to phagocytic challenge and adversely affects the response to all bacteria tested. There is no baseline activation of leukocytes by the water-soluble media, although there was some inhibition (rather than activation) of leukocyte metabolic activity. The effect of the water-soluble media in bacteria was more complex (although inhibition is minor compared to barium). Our data demonstrate that barium is a significat activator of phagocytic cells, which results in deactivation of phagocytic response when challenged; these dsata serve to explain the enhanced adverse effect of barium in cased of fecal peritonitis. (orig.)

  19. Imaging Cytometry of Human Leukocytes with Third Harmonic Generation Microscopy

    Science.gov (United States)

    Wu, Cheng-Ham; Wang, Tzung-Dau; Hsieh, Chia-Hung; Huang, Shih-Hung; Lin, Jong-Wei; Hsu, Szu-Chun; Wu, Hau-Tieng; Wu, Yao-Ming; Liu, Tzu-Ming

    2016-11-01

    Based on third-harmonic-generation (THG) microscopy and a k-means clustering algorithm, we developed a label-free imaging cytometry method to differentiate and determine the types of human leukocytes. According to the size and average intensity of cells in THG images, in a two-dimensional scatter plot, the neutrophils, monocytes, and lymphocytes in peripheral blood samples from healthy volunteers were clustered into three differentiable groups. Using these features in THG images, we could count the number of each of the three leukocyte types both in vitro and in vivo. The THG imaging-based counting results agreed well with conventional blood count results. In the future, we believe that the combination of this THG microscopy-based imaging cytometry approach with advanced texture analysis of sub-cellular features can differentiate and count more types of blood cells with smaller quantities of blood.

  20. Segmentation of leukocytes and erythrocytes in blood smear images.

    Science.gov (United States)

    Bergen, Tobias; Steckhan, Dirk; Wittenberg, Thomas; Zerfass, Thorsten

    2008-01-01

    Differential blood count is a standard method in hematological laboratory diagnosis. In the course of developing a computer-assisted microscopy system for the generation of differential blood counts, the detection and segmentation of white and red blood cells forms an essential step and its exactness is a fundamental prerequisite for the effectiveness of the subsequent classification step. We propose a method for the exact segmentation of leukocytes and erythrocytes in a simultaneous and cooperative way. We combine pixel-wise classification with template matching to locate erythrocytes and use a level-set approach in order to get the exact cell contours of leukocyte nucleus and plasma regions as well as erythrocyte regions. An evaluation comparing the performance of the algorithm to the manual segmentation performed by several persons yielded good results.

  1. Leukocyte Trafficking to the Small Intestine and Colon.

    Science.gov (United States)

    Habtezion, Aida; Nguyen, Linh P; Hadeiba, Husein; Butcher, Eugene C

    2016-02-01

    Leukocyte trafficking to the small and large intestines is tightly controlled to maintain intestinal immune homeostasis, mediate immune responses, and regulate inflammation. A wide array of chemoattractants, chemoattractant receptors, and adhesion molecules expressed by leukocytes, mucosal endothelium, epithelium, and stromal cells controls leukocyte recruitment and microenvironmental localization in intestine and in the gut-associated lymphoid tissues (GALTs). Naive lymphocytes traffic to the gut-draining mesenteric lymph nodes where they undergo antigen-induced activation and priming; these processes determine their memory/effector phenotypes and imprint them with the capacity to migrate via the lymph and blood to the intestines. Mechanisms of T-cell recruitment to GALT and of T cells and plasmablasts to the small intestine are well described. Recent advances include the discovery of an unexpected role for lectin CD22 as a B-cell homing receptor GALT, and identification of the orphan G-protein-coupled receptor 15 (GPR15) as a T-cell chemoattractant/trafficking receptor for the colon. GPR15 decorates distinct subsets of T cells in mice and humans, a difference in species that could affect translation of the results of mouse colitis models to humans. Clinical studies with antibodies to integrin α4β7 and its vascular ligand mucosal vascular addressin cell adhesion molecule 1 are proving the value of lymphocyte trafficking mechanisms as therapeutic targets for inflammatory bowel diseases. In contrast to lymphocytes, cells of the innate immune system express adhesion and chemoattractant receptors that allow them to migrate directly to effector tissue sites during inflammation. We review the mechanisms for innate and adaptive leukocyte localization to the intestinal tract and GALT, and discuss their relevance to human intestinal homeostasis and inflammation.

  2. Influence of light sources on the migration of polymorphonuclear leukocytes

    Science.gov (United States)

    DellaVecchia, Michael A.; Beard, Richard B.; Dai, Xiaoyan

    1995-05-01

    In the process of inflammation, leukocytes must travel from the intraluminal space of the capillary to the interstitial space in order to reach the site of the inflammation. The two major populations of mature human leukocytes based on the morphology are the polymorphonuclear leukocytes (PMN), and mononuclear leukocytes (MNL). Previous research on PMNs and MNLs at the Biomedical Engineering and Science Institute of Drexel University have shown that their migration can be markedly enhanced by excitation with electric and magnetic fields. This presentation demonstrates that the migration of PMNs under excitation of photons is enhanced in the red light region of (lambda) equals 660 nm and inhibited in the green light region of (lambda) equals 565 nm. There is an intensity threshold at which red light enhances migration and an intensity threshold at which green light inhibits migration. In these experiments the Boyden technique was used with the distance of the cell migration through a cellulose filter measured in terms of the leading edge. The comparison of the relative value of the distance to cell migration under a light to cell migration without a light stimulus was recorded as a cytokinetic index, K.I.. K.I. is a measure of the cytokinesis which is the progress of the cell movement in which the migration is enhanced by substances in the cell environment irrespective of a concentration gradient. The cytotactic index is a measure of cytotaxis which is the directional movement along a chemical gradient formed by a chemotactic factor. A Russian pulsed commercial laser biostimulator in the near infrared wavelength above an intensity threshold enhances PMN migration. Intermittent green and red stimulators below the intensity threshold markedly influence the cytokinetic index of PMNs while above the intensity threshold, this influence is deminished.

  3. Myxoma and vaccinia viruses bind differentially to human leukocytes.

    Science.gov (United States)

    Chan, Winnie M; Bartee, Eric C; Moreb, Jan S; Dower, Ken; Connor, John H; McFadden, Grant

    2013-04-01

    Myxoma virus (MYXV) and vaccinia virus (VACV), two distinct members of the family Poxviridae, are both currently being developed as oncolytic virotherapeutic agents. Recent studies have demonstrated that ex vivo treatment with MYXV can selectively recognize and kill contaminating cancerous cells from autologous bone marrow transplants without perturbing the engraftment of normal CD34(+) hematopoietic stem and progenitor cells. However, the mechanism(s) by which MYXV specifically recognizes and eliminates the cancer cells in the autografts is not understood. While little is known about the cellular attachment factor(s) exploited by MYXV for entry into any target cells, VACV has been shown to utilize cell surface glycosaminoglycans such as heparan sulfate (HS), the extracellular matrix protein laminin, and/or integrin β1. We have constructed MYXV and VACV virions tagged with the Venus fluorescent protein and compared their characteristics of binding to various human cancer cell lines as well as to primary human leukocytes. We report that the binding of MYXV or VACV to some adherent cell lines could be partially inhibited by heparin, but laminin blocked only VACV binding. In contrast to cultured fibroblasts, the binding of MYXV and VACV to a wide spectrum of primary human leukocytes could not be competed by either HS or laminin. Additionally, MYXV and VACV exhibited very different binding characteristics against certain select human leukocytes, suggesting that the two poxviruses utilize different cell surface determinants for the attachment to these cells. These results indicate that VACV and MYXV can exhibit very different oncolytic tropisms against some cancerous human leukocytes.

  4. Dimensions of religious involvement and leukocyte telomere length.

    Science.gov (United States)

    Hill, Terrence D; Ellison, Christopher G; Burdette, Amy M; Taylor, John; Friedman, Katherine L

    2016-08-01

    Although numerous studies suggest that religious involvement is associated with a wide range of favorable health outcomes, it is unclear whether this general pattern extends to cellular aging. In this paper, we tested whether leukocyte telomere length varies according to several dimensions of religious involvement. We used cross-sectional data from the Nashville Stress and Health Study (2011-2014), a large probability sample of 1252 black and white adults aged 22 to 69 living in Davidson County, TN, USA. Leukocyte telomere length was measured using the monochrome multiplex quantitative polymerase chain reaction method with albumin as the single-copy reference sequence. Dimensions of religious involvement included religiosity, religious support, and religious coping. Our multivariate analyses showed that religiosity (an index of religious attendance, prayer frequency, and religious identity) was positively associated with leukocyte telomere length, even with adjustments for religious support, religious coping, age, gender, race, education, employment status, income, financial strain, stressful life events, marital status, family support, friend support, depressive symptoms, smoking, heavy drinking, and allostatic load. Unlike religiosity, religious support and religious coping were unrelated to leukocyte telomere length across models. Depressive symptoms, smoking, heavy drinking, and allostatic load failed to explain any of the association between religiosity and telomere length. To our knowledge, this is the first population-based study to link religious involvement and cellular aging. Although our data suggest that adults who frequently attend religious services, pray with regularity, and consider themselves to be religious tend to exhibit longer telomeres than those who attend and pray less frequently and do not consider themselves to be religious, additional research is needed to establish the mechanisms underlying this association.

  5. Indium-111 leukocyte localization in infected prosthetic graft

    Energy Technology Data Exchange (ETDEWEB)

    Purnell, G.L.; Walker, C.W.; Allison, J.W.; Dalrymple, G.V. (Univ. of Arkansas for Medical Sciences, Little Rock (USA))

    1990-08-01

    Infective endocarditis can be difficult to prove, even in the face of strong clinical suspicion. A case in which standard methods of diagnosis failed to demonstrate endocarditis in a patient with recurrent Staphylococcus aureus bacteremia and porcine aortic valve is reported. An In-111 labelled leukocyte SPECT study demonstrated uptake in the aortic root and leaflets, and autopsy demonstrated vegetations on the leaflets. In-111 may prove useful in demonstrating endocarditis in patients with prosthetic valve infection.

  6. Leukocyte adhesion defect type 1 presenting with recurrent pyoderma gangrenosum

    Directory of Open Access Journals (Sweden)

    Neha Thakur

    2013-01-01

    Full Text Available Leukocyte adhesion deficiency 1 (LAD-1 is a rare autosomal recessive disorder of leukocyte function. LAD-1 affects about 1 per 10 million individuals and is characterized by recurrent bacterial and fungal infections and depressed inflammatory responses despite striking blood neutrophilia. Patients with the severe clinical form of LAD-1 express <0.3% of the normal amount of the β2 -integrin molecules, whereas patients with the moderate phenotype may express 2-7%. Skin infection may progress to large chronic ulcers with polymicrobial infection, including anaerobic organisms. The ulcers heal slowly, require months of antibiotic treatment, and often require plastic surgical grafting. The diagnosis of LAD-1 is established most readily by flow cytometric measurements of surface CD11b in stimulated and unstimulated neutrophils using monoclonal antibodies directed against CD11b. Pyoderma gangrenosum (PG is an uncommon condition characterized by recurrent sterile, inflammatory skin ulcers. Commonly, PG occurs in the context of inflammatory bowel disease or rheumatic, hematologic, or immunologic disorders. Here, we present a 5-year-old female with a long history of PG, which healed with atrophic scarring, who was ultimately diagnosed with leukocyte adhesion deficiency type 1 (LAD1. She had a good response to high-dose prednisone therapy (2 mg/kg and was discharged after 3 weeks of admission but only to be re-admitted 3 weeks later with severe pneumonia. During hospital stay, she developed pneumothorax and pneumomediastinum and later succumbed to her illness.

  7. Tracking flow of leukocytes in blood for drug analysis

    Science.gov (United States)

    Basharat, Arslan; Turner, Wesley; Stephens, Gillian; Badillo, Benjamin; Lumpkin, Rick; Andre, Patrick; Perera, Amitha

    2011-03-01

    Modern microscopy techniques allow imaging of circulating blood components under vascular flow conditions. The resulting video sequences provide unique insights into the behavior of blood cells within the vasculature and can be used as a method to monitor and quantitate the recruitment of inflammatory cells at sites of vascular injury/ inflammation and potentially serve as a pharmacodynamic biomarker, helping screen new therapies and individualize dose and combinations of drugs. However, manual analysis of these video sequences is intractable, requiring hours per 400 second video clip. In this paper, we present an automated technique to analyze the behavior and recruitment of human leukocytes in whole blood under physiological conditions of shear through a simple multi-channel fluorescence microscope in real-time. This technique detects and tracks the recruitment of leukocytes to a bioactive surface coated on a flow chamber. Rolling cells (cells which partially bind to the bioactive matrix) are detected counted, and have their velocity measured and graphed. The challenges here include: high cell density, appearance similarity, and low (1Hz) frame rate. Our approach performs frame differencing based motion segmentation, track initialization and online tracking of individual leukocytes.

  8. Does human leukocyte elastase degrade intact skin elastin?

    Science.gov (United States)

    Schmelzer, Christian E H; Jung, Michael C; Wohlrab, Johannes; Neubert, Reinhard H H; Heinz, Andrea

    2012-11-01

    This study aimed to investigate the susceptibility of intact fibrillar human elastin to human leukocyte elastase and cathepsin G. Elastin is a vital protein of the extracellular matrix of vertebrates, and provides exceptional properties including elasticity and tensile strength to many tissues and organs, including the aorta, lung, cartilage, elastic ligaments and skin, and is thus critical for their long-term function. Mature elastin is an insoluble and extremely durable protein that undergoes very little turnover, but sustained exposure to proteases may lead to irreversible and severe damage, and thus to functional loss of the elastic fiber network. Hence, it is a key issue to understand which enzymes actually initiate elastolysis under certain pathological conditions or during intrinsic aging. In this paper, we provide a complete workflow for isolation of pure and intact elastin from very small tissue samples to test enzymes for their elastolytic potential. This workflow was applied to skin samples from variously aged individuals, and it was found that strong differences exist in the degradability of the elastins investigated. In summary, human leukocyte elastase was unable to degrade intact elastin fibers but hydrolyzed elastin derived from the skin of old people. However, cathepsin G cleaved all elastin samples, even those derived from younger individuals. These results indicate that human leukocyte elastase is not a driving force for elastolysis, but may nevertheless promote further breakdown of elastic fibers after the action of other enzymes such as cathepsin G. © 2012 The Authors Journal compilation © 2012 FEBS.

  9. Comparison of photonic and electromagnetic effects on the human leukocyte

    Science.gov (United States)

    DellaVecchia, Michael A.; Beard, Richard B.; Feng, D.; Dai, Xiaoyan; Pourrezaei, Kambiz; Priezzhev, Alexander V.

    1998-06-01

    The dielectric and magnetic influence on human cells have been widely studied previously by the authors. Recently, the effects of energy in the visible electromagnetic spectrum have been investigated. In this subsequent study, the photonic effects on the in vitro migration of the polymorphonuclear and mononuclear leukocytes are compared with the corresponding electromagnetic field effects. Dielectric spectra of the polymorph in the 300 KHz to 400 KHz and 700 KHz to 800 KHz range have been measured. At frequencies of 350 KHz and 720 KHz an increase in the migration of the polymorphonuclear leukocyte have been observed. This stimulation was attributed to the charges on the nuclear surface. Recent preliminary data have shown a similar increased migration in the 20 MHz range. Photonic studies have indicated an enhanced migration for the polymorphonuclear leukocytes at a wavelength of 660 nm (red) and an inhibited migration at 565 nm (green). The photonic effects were postulated to be the results of a biochemical interaction rather than a membranous surface charge displacement secondary to an electric field. The migration of the white blood cells were measurement via the Boyden chamber technique and expressed in terms of a cytokinetic index which expresses the cellular movement independent of its environmental concentration gradient.

  10. Common leukocyte antigen staining of a primitive sarcoma.

    Science.gov (United States)

    McDonnell, J M; Beschorner, W E; Kuhajda, F P; deMent, S H

    1987-04-15

    A 4-year-old boy presented with symptoms of tracheal obstruction and was found to have a polypoid tracheal mass, which was studied by biopsy. Light microscopy showed a tumor composed of small cells with round to oval dark nuclei, clumped chromatin, one to two nucleoli, and small, variable amounts of indistinct pink cytoplasm. In other areas the tumor had a loose, spindle appearance, with some cells showing more elongated nuclei, and fibrillar pink cytoplasm consistent with strap cells. Cross striations were not found. Electron microscopy showed desmosomes and 7 to 10 nm cytoplasmic filaments forming dense bodies. The findings are most consistent with a primitive sarcoma, probably rhabdomyosarcoma. Immunoperoxidase with three monoclonal antibodies for common leukocyte antigen showed diffuse membraneous staining with fresh-frozen tissue. All other lymphocyte and monocyte marker studies were negative. We believe that this case of anticommon leukocyte antigen staining, a rhabdomyosarcoma, represents the first report of a false positive reaction with monoclonal antibody to common leukocyte antigen.

  11. Macrophage recognition of ICAM-3 on apoptotic leukocytes.

    Science.gov (United States)

    Moffatt, O D; Devitt, A; Bell, E D; Simmons, D L; Gregory, C D

    1999-06-01

    Cells undergoing apoptosis are cleared rapidly by phagocytes, thus preventing tissue damage caused by loss of plasma membrane integrity. In this study, we show that the surface of leukocytes is altered during apoptosis such that the first Ig-like domain of ICAM-3 (CD50) can participate in the recognition and phagocytosis of the apoptotic cells by macrophages. Macrophage recognition of apoptotic cell-associated ICAM-3 was demonstrated both on leukocytes and, following transfection of exogenous ICAM-3, on nonleukocytes. The change in ICAM-3 was a consistent consequence of apoptosis triggered by various stimuli, suggesting that it occurs as part of a final common pathway of apoptosis. Alteration of ICAM-3 on apoptotic cells permitting recognition by macrophages resulted in a switch in ICAM-3-binding preference from the prototypic ICAM-3 counterreceptor, LFA-1, to an alternative macrophage receptor. Using mAbs to block macrophage/apoptotic cell interactions, we were unable to obtain evidence that either the alternative ICAM-3 counterreceptor alpha d beta 2 or the apoptotic cell receptor alpha v beta 3 was involved in the recognition of ICAM-3. By contrast, mAb blockade of macrophage CD14 inhibited ICAM-3-dependent recognition of apoptotic cells. These results show that ICAM-3 can function as a phagocytic marker of apoptotic leukocytes on which it acquires altered macrophage receptor-binding activity.

  12. Increased oxidative DNA damage in mononuclear leukocytes in vitiligo

    Energy Technology Data Exchange (ETDEWEB)

    Giovannelli, Lisa [Department of Preclinical and Clinical Pharmacology, University of Florence, Viale Pieraccini 6, 50139 Florence (Italy)]. E-mail: lisag@pharm.unifi.it; Bellandi, Serena [Department of Dermatological Sciences, University of Florence, Viale Pieraccini 6, 50139 Florence (Italy); Pitozzi, Vanessa [Department of Preclinical and Clinical Pharmacology, University of Florence, Viale Pieraccini 6, 50139 Florence (Italy); Fabbri, Paolo [Department of Dermatological Sciences, University of Florence, Viale Pieraccini 6, 50139 Florence (Italy); Dolara, Piero [Department of Preclinical and Clinical Pharmacology, University of Florence, Viale Pieraccini 6, 50139 Florence (Italy); Moretti, Silvia [Department of Dermatological Sciences, University of Florence, Viale Pieraccini 6, 50139 Florence (Italy)

    2004-11-22

    Vitiligo is an acquired pigmentary disorder of the skin of unknown aetiology. The autocytotoxic hypothesis suggests that melanocyte impairment could be related to increased oxidative stress. Evidences have been reported that in vitiligo oxidative stress might also be present systemically. We used the comet assay (single cell alkaline gel electrophoresis) to evaluate DNA strand breaks and DNA base oxidation, measured as formamidopyrimidine DNA glycosylase (FPG)-sensitive sites, in peripheral blood cells from patients with active vitiligo and healthy controls. The basal level of oxidative DNA damage in mononuclear leukocytes was increased in vitiligo compared to normal subjects, whereas DNA strand breaks (SBs) were not changed. This alteration was not accompanied by a different capability to respond to in vitro oxidative challenge. No differences in the basal levels of DNA damage in polymorphonuclear leukocytes were found between patients and healthy subjects. Thus, this study supports the hypothesis that in vitiligo a systemic oxidative stress exists, and demonstrates for the first time the presence of oxidative alterations at the nuclear level. The increase in oxidative DNA damage shown in the mononuclear component of peripheral blood leukocytes from vitiligo patients was not particularly severe. However, these findings support an adjuvant role of antioxidant treatment in vitiligo.

  13. Peripheral blood leukocyte count as an index of defense status in the leukopenic host

    Energy Technology Data Exchange (ETDEWEB)

    Cawley, S.; Findon, G.; Miller, T.E.

    1988-07-01

    These experimental studies have investigated the reliability of the peripheral blood leukocyte count to predict whether the leukopenic host can contain or eliminate infection. Additionally, we have investigated the possibility that determination of leukocyte recruitment, supplementary to peripheral blood leukocyte counts, might allow individuals with neutropenia at risk from serious infection to be distinguished with greater certainty. Varying doses of radiation, cyclophosphamide, and methylprednisolone were used to induce distinct levels of leukopenia in rats. Leukocyte recruitment was measured by quantifying the response of neutropenic animals to evocative, subcutaneous stimuli, and the results of this assay were then compared with circulating leukocyte counts in the same individuals. Six models of experimentally induced infection were used to compare circulating and recruitable leukocytes as indicators of the susceptibility of the leukopenic host to infection. Response curves relating leukocyte numbers to host resistance were similar when circulating or recruitable leukocytes were used as an index of defense capability. These findings support the use of peripheral blood leukocyte numbers as an index of resistance to infection in individuals with leukopenia and suggest that functional analyses such as leukocyte recruitment are unlikely to provide additional information.

  14. Leukocyte Mitochondrial DNA Alteration in Systemic Lupus Erythematosus and Its Relevance to the Susceptibility to Lupus Nephritis

    Directory of Open Access Journals (Sweden)

    Yau-Huei Wei

    2012-07-01

    Full Text Available The role of mitochondrial DNA (mtDNA alterations in the pathophysiology of systemic lupus erythematosus (SLE remains unclear. We investigated sequence variations in the D310 region and copy number change of mtDNA in 85 SLE patients and 45 normal subjects. Leukocyte DNA and RNA were extracted from leukocytes of the peripheral venous blood. The D310 sequence variations and copy number of mtDNA, and mRNA expression levels of mtDNA-encoded genes in leukocytes were determined by quantitative real-time polymerase chain reaction (Q-PCR and PCR-based direct sequencing, respectively. We found that leukocyte mtDNA in SLE patients exhibited higher frequency of D310 heteroplasmy (69.4% vs. 48.9%, p = 0.022 and more D310 variants (2.2 vs. 1.7, p = 0.014 than those found in controls. Among normal controls and patients with low, medium or high SLE disease activity index (SLEDAI, an ever-increasing frequency of D310 heteroplasmy was observed (p = 0.021. Leukocyte mtDNA copy number tended to be low in patients of high SLEDAI group (p = 0.068, especially in those harboring mtDNA with D310 heteroplasmy (p = 0.020. Moreover, the mtDNA copy number was positively correlated with the mRNA level of mtDNA-encoded ND1 (NADH dehydrogenase subunit 1 (p = 0.041 and ATPase 6 (ATP synthase subunit 6 (p = 0.030 genes. Patients with more D310 variants were more susceptible to lupus nephritis (p = 0.035. Taken together, our findings suggest that decrease in the mtDNA copy number and increase in D310 heteroplasmy of mtDNA are related to the development and progression of SLE, and that the patients harboring more D310 variants of mtDNA are more susceptible to lupus nephritis.

  15. Importance of Primary Capture and L-Selectin–Dependent Secondary Capture in Leukocyte Accumulation in Inflammation and Atherosclerosis in Vivo

    OpenAIRE

    2001-01-01

    In the multistep process of leukocyte extravasation, the mechanisms by which leukocytes establish the initial contact with the endothelium are unclear. In parallel, there is a controversy regarding the role for L-selectin in leukocyte recruitment. Here, using intravital microscopy in the mouse, we investigated leukocyte capture from the free flow directly to the endothelium (primary capture), and capture mediated through interactions with rolling leukocytes (secondary capture) in venules, in ...

  16. Biomimetic Leukocyte Adhesion: A Review of Microfluidic and Computational Approaches and Applications

    Institute of Scientific and Technical Information of China (English)

    J.Hanzlik; E.Cretekos; K.A.Lamkin-Kennard

    2008-01-01

    Leukocyte rolling and adhesion are complex physiological processes that have received a great deal of attention over the past decade. Significant increases in the knowledge base related to how leukocytes adhere in shear flows have occurred as a result of the development of novel experimental and computational techniques. Micro- and nano-fabrication techniques have enabled the development of novel flow devices for studying leukocyte adhesion in simple and complex geometries. Improve-ments in computer technology have enabled simulations of complex flow processes to be developed. As a result of these ad-vances in knowledge related to leukocyte adhesion, numerous novel devices have been developed that mimic the leukocyte rolling and adhesion process. Examples of these devices include cell separation and enrichment devices and targeted ultrasound contrast agents. Future advances related to leukocyte rolling and adhesion processes hold great promise for advancing our knowledge of disease processes as well as development of novel therapeutic devices.

  17. Assessment of malathion and its effects on leukocytes in human blood samples

    Science.gov (United States)

    Sharma, Amit Kumar; Tiwari, Udita; Gaur, Mulayam Singh; Tiwari, Rajeev Kumar

    2016-01-01

    Abstract In the present paper, we report a reproducible, cost effective, fast response method for detection of malathion and its effects on leukocytes in different human blood groups. Spectroscopic methods (UV-Vis spectrometry) and Fourier transform infrared coupled with solid phase extraction were applied for analyzing malathion content in human blood plasma. The spiking levels of malathion in the range of 0.1-1.7 µg/mL were extracted from blood plasma samples using SPE. The present active functional groups (C = O; P-O-C; -OH; P = S) were also characterized. The recovery rate of malathion was 80%±4.5%. The calculated correlation coefficient was 0.9799, indicating the linearity of the results. The limit of detection (LOD) and limit of quantification (LOQ) were (0.1-1.7) µg/mL and (0.3-1.5) µg/mL, respectively. Malathion <1.0 µg/mL showed no significant change while higher levels of malathion exposure (1.5 µg/mL and 3.0 µg/mL) reduced the number of white blood cells. In conclusion, the spectroscopic results may be useful to understand the mechanism of other pesticides such as methyl parathion and parathion.

  18. Role of Gallium and labeled leukocyte scintigraphy in AIDS patient

    Energy Technology Data Exchange (ETDEWEB)

    Palestro, C.J. [Division of nuclear medicine, Long Island Jewish Medical Center, New Hyde Park, New York (United States); Goldsmith, S.J. [Division of nuclear medicine, New York Hospital, Cornell Medical Center, New York (United States)

    1995-09-01

    Because AIDS patients frequently present with minimal symptomatology, radionuclide imaging with its ability to survey the entire body, is especially valuable. Gallium-67 citrate, the most commonly performed radionuclide study for localizing infection in these patients, is most useful for detecting opportunistic infections, especially in the thorax. A negative gallium scan, particularly when the chest X-ray is unremarkable, rules strongly against pulmonary disease. A negative gallium scan in a patient with an abnormal chest X-ray and Kaposi`s sarcoma, suggests that the patient`s respiratory distress is related to the neoplasm. Diffuse pulmonary parenchymal uptake of gallium in the HIV (+) patient is most often associated with PCP. While there are other causes of diffuse pulmonary uptake, the more intense or heterogeneous the uptake, the more likely the patient is to have PCP. Focal pulmonary uptake is usually associated with bacterial pneumonia although PCP may occasionally present in this fashion. Lymph node uptake of gallium is usually associated with Mycob acterium avium complex, tuberculosis, or Iymphoma. When corresponding abnormalities are present on thallium scintigraphy lymphoma is likely. Gallium positive, thallium negative, studies suggest mycobacterial disease. Labeled leukocyte imaging is not useful for detecting opportunistic infections probably because of the inflammatory response incited by these organisms. Leukocyte imaging is, however, more sensitive for detecting bacterial pneumonia. In the abdomen, gallium imaging is most useful for identifying lymphadenopathy, while labeled leukocyte imaging is superior for detecting AlDS-associated colitides. In summary, radionuclide studies are valuable diagnostic modalities in AIDS. Their success can be maximized by tailoring the study to the individual`s needs.

  19. Imaging of leukocyte trafficking in Alzheimer’s disease

    Directory of Open Access Journals (Sweden)

    Gabriela eConstantin

    2016-02-01

    Full Text Available Alzheimer’s disease (AD is the most common neurodegenerative disorder and is characterized by a progressive decline of cognitive functions. The neuropathological features of AD include amyloid beta (Aβ deposition, intracellular neurofibrillary tangles derived from the cytoskeletal hyperphosphorylated tau protein, amyloid angiopathy, the loss of synapses, and neuronal degeneration. In the last decade, inflammation has emerged as a key feature of AD, but most studies have focused on the role of microglia-driven neuroinflammation mechanisms. A dysfunctional blood–brain barrier (BBB has also been implicated in the pathogenesis of AD, and several studies have demonstrated that the vascular deposition of Aβ induces the expression of adhesion molecules and alters the expression of tight junction proteins, potentially facilitating the transmigration of circulating leukocytes. Two-photon laser scanning microscopy (TPLSM has become an indispensable tool to dissect the molecular mechanisms controlling leukocyte trafficking in the central nervous system (CNS. Recent TPLSM studies have shown that vascular deposition of Aβ in the CNS promotes intraluminal neutrophil adhesion and crawling on the brain endothelium, and also that neutrophils extravasate in the parenchyma preferentially in areas with Aβ deposits. These studies have also highlighted a role for LFA-1 integrin in neutrophil accumulation in the CNS of AD-like disease models, revealing that LFA-1 inhibition reduces the corresponding cognitive deficit and AD neuropathology. In this article, we consider how current imaging techniques can help to unravel new inflammation mechanisms in the pathogenesis of AD and identify novel therapeutic strategies to treat the disease by interfering with leukocyte trafficking mechanisms.

  20. The Alpha-Defensin Immunoassay and Leukocyte Esterase Colorimetric Strip Test for the Diagnosis of Periprosthetic Infection

    Science.gov (United States)

    Wyatt, M.C.; Beswick, A.D.; Kunutsor, S.K.; Wilson, M.J.; Whitehouse, M.R.; Blom, A.W.

    2016-01-01

    Background: Synovial biomarkers have recently been adopted as diagnostic tools for periprosthetic joint infection (PJI), but their utility is uncertain. The purpose of this systematic review and meta-analysis was to synthesize the evidence on the accuracy of the alpha-defensin immunoassay and leukocyte esterase colorimetric strip test for the diagnosis of PJI compared with the Musculoskeletal Infection Society diagnostic criteria. Methods: We performed a systematic review to identify diagnostic technique studies evaluating the accuracy of alpha-defensin or leukocyte esterase in the diagnosis of PJI. MEDLINE and Embase on Ovid, ACM, ADS, arXiv, CERN DS (Conseil Européen pour la Recherche Nucléaire Document Server), CrossRef DOI (Digital Object Identifier), DBLP (Digital Bibliography & Library Project), Espacenet, Google Scholar, Gutenberg, HighWire, IEEE Xplore (Institute of Electrical and Electronics Engineers digital library), INSPIRE, JSTOR (Journal Storage), OAlster (Open Archives Initiative Protocol for Metadata Harvesting), Open Content, Pubget, PubMed, and Web of Science were searched for appropriate studies indexed from inception until May 30, 2015, along with unpublished or gray literature. The classification of studies and data extraction were performed independently by 2 reviewers. Data extraction permitted meta-analysis of sensitivity and specificity with construction of receiver operating characteristic curves for each test. Results: We included 11 eligible studies. The pooled diagnostic sensitivity and specificity of alpha-defensin (6 studies) for PJI were 1.00 (95% confidence interval [CI], 0.82 to 1.00) and 0.96 (95% CI, 0.89 to 0.99), respectively. The area under the curve (AUC) for alpha-defensin and PJI was 0.99 (95% CI, 0.98 to 1.00). The pooled diagnostic sensitivity and specificity of leukocyte esterase (5 studies) for PJI were 0.81 (95% CI, 0.49 to 0.95) and 0.97 (95% CI, 0.82 to 0.99), respectively. The AUC for leukocyte esterase and PJI

  1. Cell-fluid Interaction: Coupling Between the Deformation of an Adherent Leukocyte and the Shear Flow

    Institute of Scientific and Technical Information of China (English)

    2005-01-01

    1 IntroductionLeukocyte adhesion is a natural physiopathological phenomenon and the balance between the hemodynamic forces and adhesion forces (molecular bonds) plays a key role. According to hemodynamic theories, blood flow induces the change of the shape and the spatial arrangement of leukocytes. These changes may in turn induce the redistribution of blood flow around the cell. Therefore there exist interaction of the adherent leukocyte with blood flow, which is called as the coupling between the hemodyna...

  2. Cystine accumulation and clearance by normal and cystinotic leukocytes exposed to cystine dimethyl ester.

    OpenAIRE

    Steinherz, R; Tietze, F.; Gahl, W A; Triche, T J; Chiang, H.; Modesti, A.; Schulman, J D

    1982-01-01

    Upon exposure to 0.25 mM cystine dimethyl ester, normal and cystinotic leukocytes accumulate substantially more intracellular cystine than is present endogenously in cystinotic cells. Leukocytes loaded by exposure to cystine dimethyl ester may have abnormally lucent and distended lysosomes, and the cystine is compartmentalized within the granular fraction of the cells. After the cells are exposed to cystine dimethyl ester, cystine clearance from normal leukocytes is much faster than from cyst...

  3. Osteomyelitis in leukocyte adhesion deficiency type 1 syndrome

    DEFF Research Database (Denmark)

    Jabbari Azad, Farahzad; Ardalan, Maryam; H.Rafati, Ali;

    2010-01-01

    Leukocyte adhesion deficiency type 1 (LAD-1) is a rare, inherited immunodeficiency that affects one per million people yearly and usually presents with recurrent, indolent bacterial infections of the skin, mouth, and respiratory tract and impaired pus formation and wound healing. A 13-year-old gi......(10.60%). A plain radiography of the left leg revealed osteomyelitis. It is highly suggested that patients diagnosed mild to moderate LAD-1 with recurrent skin infection and simultaneous weak response to conventional therapy undergo (BMT) marrow transplant to prohibit subsequent life...

  4. The Many Faces of Human Leukocyte Antigen-G

    DEFF Research Database (Denmark)

    Dahl, Mette; Djurisic, Snezana; Hviid, Thomas Vauvert F

    2014-01-01

    is the human leukocyte antigen (HLA)-G, a nonclassical HLA protein displaying limited polymorphism, restricted tissue distribution, and a unique alternative splice pattern. HLA-G is primarily expressed in placenta and plays multifaceted roles during pregnancy, both as a soluble and a membrane-bound molecule...... pregnancy and pregnancy complications, such as preeclampsia, recurrent spontaneous abortions, and subfertility or infertility. This review aims to clarify the multifunctional role of HLA-G in pregnancy-related disorders by focusing on genetic variation, differences in mRNA stability between HLA-G alleles...

  5. Theileria-transformed bovine leukocytes have cancer hallmarks.

    Science.gov (United States)

    Tretina, Kyle; Gotia, Hanzel T; Mann, David J; Silva, Joana C

    2015-07-01

    The genus Theileria includes tick-transmitted apicomplexan parasites of ruminants with substantial economic impact in endemic countries. Some species, including Theileria parva and Theileria annulata, infect leukocytes where they induce phenotypes that are shared with some cancers, most notably immortalization, hyperproliferation, and dissemination. Despite considerable research into the affected host signaling pathways, the parasite proteins directly responsible for these host phenotypes remain unknown. In this review we outline current knowledge on the manipulation of host cells by transformation-inducing Theileria, and we propose that comparisons between cancer biology and host-Theileria interactions can reveal chemotherapeutic targets against Theileria-induced pathogenesis based on cancer treatment approaches.

  6. Longitudinal evaluation of leukocyte transcripts in killer whales (Orcinus Orca).

    Science.gov (United States)

    Sitt, Tatjana; Bowen, Lizabeth; Lee, Chia-Shan; Blanchard, Myra T; McBain, James; Dold, Christopher; Stott, Jeffrey L

    2016-07-01

    Early identification of illness and/or presence of environmental and/or social stressors in free-ranging and domestic cetaceans is a priority for marine mammal health care professionals. Incorporation of leukocyte gene transcript analysis into the diagnostic tool kit has the potential to augment classical diagnostics based upon ease of sample storage and shipment, inducible nature and well-defined roles of transcription and associated downstream actions. Development of biomarkers that could serve to identify "insults" and potentially differentiate disease etiology would be of great diagnostic value. To this end, a modest number of peripheral blood leukocyte gene transcripts were selected for application to a domestic killer whale population with a focus on broad representation of inducible immunologically relevant genes. Normalized leukocyte transcript values, longitudinally acquired from 232 blood samples derived from 26 clinically healthy whales, were not visibly influenced temporally nor by sex or the specific Park in which they resided. Stability in leukocyte transcript number during periods of health enhances their potential use in diagnostics through identification of outliers. Transcript levels of two cytokine genes, IL-4 and IL-17, were highly variable within the group as compared to the other transcripts. IL-4 transcripts were typically absent. Analysis of transcript levels on the other genes of interest, on an individual animal basis, identified more outliers than were visible when analyzed in the context of the entire population. The majority of outliers (9 samples) were low, though elevated transcripts were identified for IL-17 from 2 animals and one each for Cox-2 and IL-10. The low number of outliers was not unexpected as sample selection was intentionally directed towards animals that were clinically healthy at the time of collection. Outliers may reflect animals experiencing subclinical disease that is transient and self-limiting. The immunologic

  7. Osteomyelitis in leukocyte adhesion deficiency type 1 syndrome

    DEFF Research Database (Denmark)

    Jabbari Azad, Farahzad; Ardalan, Maryam; H.Rafati, Ali

    2010-01-01

    Leukocyte adhesion deficiency type 1 (LAD-1) is a rare, inherited immunodeficiency that affects one per million people yearly and usually presents with recurrent, indolent bacterial infections of the skin, mouth, and respiratory tract and impaired pus formation and wound healing. A 13-year-old girl......(10.60%). A plain radiography of the left leg revealed osteomyelitis. It is highly suggested that patients diagnosed mild to moderate LAD-1 with recurrent skin infection and simultaneous weak response to conventional therapy undergo (BMT) marrow transplant to prohibit subsequent life...

  8. Longitudinal evaluation of leukocyte transcripts in killer whales (Orcinus Orca)

    Science.gov (United States)

    Sitt, Tatjana; Bowen, Lizabeth; Lee, Chia-Shan; Blanchard, Myra; McBain, James; Dold, Christopher; Stott, Jeffrey L.

    2016-01-01

    Early identification of illness and/or presence of environmental and/or social stressors in free-ranging and domestic cetaceans is a priority for marine mammal health care professionals. Incorporation of leukocyte gene transcript analysis into the diagnostic tool kit has the potential to augment classical diagnostics based upon ease of sample storage and shipment, inducible nature and well-defined roles of transcription and associated downstream actions. Development of biomarkers that could serve to identify “insults” and potentially differentiate disease etiology would be of great diagnostic value. To this end, a modest number of peripheral blood leukocyte gene transcripts were selected for application to a domestic killer whale population with a focus on broad representation of inducible immunologically relevant genes. Normalized leukocyte transcript values, longitudinally acquired from 232 blood samples derived from 26 clinically healthy whales, were not visibly influenced temporally nor by sex or the specific Park in which they resided. Stability in leukocyte transcript number during periods of health enhances their potential use in diagnostics through identification of outliers. Transcript levels of two cytokine genes, IL-4 and IL-17, were highly variable within the group as compared to the other transcripts. IL-4 transcripts were typically absent. Analysis of transcript levels on the other genes of interest, on an individual animal basis, identified more outliers than were visible when analyzed in the context of the entire population. The majority of outliers (9 samples) were low, though elevated transcripts were identified for IL-17 from 2 animals and one each for Cox-2 and IL-10. The low number of outliers was not unexpected as sample selection was intentionally directed towards animals that were clinically healthy at the time of collection. Outliers may reflect animals experiencing subclinical disease that is transient and self-limiting. The

  9. Correlation between Leukocyte Numbers and Body Size of Rainbow Trout

    DEFF Research Database (Denmark)

    Mohammad, Rezkar Jaafar; Otani, Maki; Kania, Per Walter

    2016-01-01

    towards an antigen to be initiated even in fry. The number of leukocytes in individual fish at different developmental stages is likely to influence the capacity of the fish to respond simultaneously to several antigens (pathogens and vaccine components). This parameter may therefore be crucial for both...... - 780 g (group IV) were investigated. The number of lymphocytes was generally higher in head kidney compared to blood and spleen but they dominated in all samples (blood, head kidney and spleen) and their numbers increased exponentially with fish size. Percentages of lymphocytes in relation...

  10. Biocompatibility and permeability of dialyzer membranes do not affect anemia, erythropoietin dosage or mortality in japanese patients on chronic non-reuse hemodialysis: a prospective cohort study from the J-DOPPS II study.

    Science.gov (United States)

    Yokoyama, Hitoshi; Kawaguchi, Takehiko; Wada, Takashi; Takahashi, Yoshinori; Higashi, Takahiro; Yamazaki, Shin; Fukuhara, Shunichi; Akiba, Takashi; Akizawa, Tadao; Asano, Yasushi; Kurokawa, Kiyoshi; Saito, Akira

    2008-01-01

    Considerable controversy exists over the impact of the biocompatibility and flux characteristics of dialyzer membranes on anemia in chronic hemodialysis patients. A subset of 1,207 subjects from the Japanese arm of DOPPS phase II was analyzed. Patient characteristics included mean age 59 years, male sex 60%, BMI 20.6, time on dialysis therapy 7.8 years, and diabetes rate 27%. Dialysis parameters were Kt/V 1.33, and normalized protein catabolic rate 1.05 g/kg/day. Initial hemoglobin level was 10.1 g/dl. 79% were treated by intravenous erythropoietin with mean weekly doses of 4,500 IU. Hemoglobin levels and erythropoietin doses during 2-year study period were not affected by dialysis membrane biocompatibility (unmodified cellulose or biocompatible) or flux (standard or high performance). The 2-year survival rate was 90.9% and was influenced by older age, presence of cardiovascular diseases and amyloidosis, lower levels of BMI and serum albumin, but not by other variables, including dialysis membranes. Use of biocompatible membranes was associated with a lower all-cause mortality (8.3 vs. 13.0% for bioincompatible, p = 0.037), but this difference was not significant in multivariate analyses (hazard ratio 0.70, p = 0.17 by Cox multivariate analysis). The biocompatibility and permeability of dialyzer membranes had no effect on anemia, erythropoietin dosage or all-cause mortality in Japanese chronic hemodialysis patients treated by non-reuse dialysis. Copyright 2008 S. Karger AG, Basel.

  11. The Prognostic Values of Leukocyte Rho Kinase Activity in Acute Ischemic Stroke

    Directory of Open Access Journals (Sweden)

    Cheng-I. Cheng

    2014-01-01

    Full Text Available Objective. It has been reported that leukocyte ROCK activity is elevated in patients after ischemic stroke, but it is unclear whether leukocyte ROCK activity is associated with clinical outcomes following acute stroke events. The objective of this study is to investigate if leukocyte ROCK activity can predict the outcomes in patients with acute ischemic stroke. Materials and Methods. We enrolled 110 patients of acute ischemic stroke and measured the leukocyte ROCK activity and plasma level of inflammatory cytokines to correlate the clinical outcomes of these patients. Results. The leukocyte ROCK activity at 48 hours after admission in acute ischemic stroke patients was higher as compared to a risk-matched population. The leukocyte ROCK activity significantly correlated with National Institute of Health Stroke Scale (NIHSS difference between admission and 90 days after stroke event. Kaplan-Meier survival estimates showed lower stroke-free survival during follow-up period in patients with high leukocyte ROCK activity or plasma hsCRP level. Leukocyte ROCK activity independently predicted the recurrent stroke in patients with atherosclerotic stroke. Conclusions. This study shows elevated leukocyte ROCK activity in patients with ischemic stroke as compared to risk-matched subjects and is an independent predictor for recurrent stroke.

  12. Spleen tyrosine kinase Syk is critical for sustained leukocyte adhesion during inflammation in vivo

    Directory of Open Access Journals (Sweden)

    Poeschl Johannes

    2007-11-01

    Full Text Available Abstract Background During inflammation, β2-integrins mediate leukocyte adhesion to the endothelium accompanied by the activation of the spleen tyrosine kinase Syk. Results We investigated leukocyte adhesion and rolling in cremaster muscle venules before and during stimulation with fMLP using mice with a Syk-/- hematopoietic system. In unstimulated venules, Syk-/- leukocytes adhered less efficiently than control leukocytes while rolling was similar between Syk-/- and control leukocytes. During fMLP-superfusion, control mice showed significantly increased adhesion accompanied by reduced rolling. For Syk-/- leukocytes, an increase in adhesion with a concomitant decrease in rolling was only observed during the first three minutes during fMLP stimulation, but not at later time points. We also investigated leukocyte spreading against the vessel wall during fMLP stimulation and found a significant impairment of spreading for Syk-/- leukocytes. Additional in vitro experiments revealed that the adhesion and spreading defect seen in Syk-/- chimeric mice was due to compromised β2-integrin-mediated outside-in signaling. Conclusion We provide substantial evidence for an important role of Syk in mediating β2-integrin dependent outside-in signaling leading to sustained leukocyte adhesion and spreading during the inflammatory response in vivo.

  13. Local histamine release increases leukocyte rolling in the cerebral microcirculation of the mouse.

    Science.gov (United States)

    Yong, T; Zheng, M Q; Linthicum, D S

    1997-10-01

    Histamine-mediated induction of leukocyte rolling and adhesion in the cerebral microcirculation was examined in two inbred strains of mice (SJL/J and BALB/c). A cranial window was surgically prepared for the visualization of the cerebral microcirculation using intra-vital microscopy. Leukocyte rolling and adhesion to pial venular walls were assessed during off-line video playback analyses. The surgical preparation of the cranial windows was found to trigger 'spontaneous' leukocyte rolling, and this was attributed to disruption of dural mast cells and localized release of vasoactive histamine. This spontaneous leukocyte rolling was observed only in the SJL/J strain of mice, and could be prevented by presurgical treatment with the mast cell stabilizer sodium cromoglycate. BALB/c mice did not show 'spontaneous' leukocyte rolling or adhesion; this strain is known to have low numbers of CNS-associated mast cells. Exogenous histamine, applied topically to the cerebral microcirculation via the cranial window in mice pretreated with sodium cromoglycate, produced significant dose-dependent increases in leukocyte rolling and adhesion to pial venules in SJL/J mice, but not in BALB/c mice. Diphenhydramine (H1 receptor antagonist), but not cimetidine (H2 receptor antagonist), abolished both 'spontaneous' and histamine-induced leukocyte rolling. Anti-P-selectin antibody was found efficiently to block both spontaneous and histamine-induced increases in leukocyte rolling, but not leukocyte adhesion.

  14. Leukocytes Breach Endothelial Barriers by Insertion of Nuclear Lobes and Disassembly of Endothelial Actin Filaments

    Directory of Open Access Journals (Sweden)

    Sagi Barzilai

    2017-01-01

    Full Text Available The endothelial cytoskeleton is a barrier for leukocyte transendothelial migration (TEM. Mononuclear and polymorphonuclear leukocytes generate gaps of similar micron-scale size when squeezing through inflamed endothelial barriers in vitro and in vivo. To elucidate how leukocytes squeeze through these barriers, we co-tracked the endothelial actin filaments and leukocyte nuclei in real time. Nuclear squeezing involved either preexistent or de novo-generated lobes inserted into the leukocyte lamellipodia. Leukocyte nuclei reversibly bent the endothelial actin stress fibers. Surprisingly, formation of both paracellular gaps and transcellular pores by squeezing leukocytes did not require Rho kinase or myosin II-mediated endothelial contractility. Electron-microscopic analysis suggested that nuclear squeezing displaced without condensing the endothelial actin filaments. Blocking endothelial actin turnover abolished leukocyte nuclear squeezing, whereas increasing actin filament density did not. We propose that leukocyte nuclei must disassemble the thin endothelial actin filaments interlaced between endothelial stress fibers in order to complete TEM.

  15. Kinetics of reversible-sequestration of leukocytes by the isolated perfused rat lung

    Energy Technology Data Exchange (ETDEWEB)

    Goliaei, B.

    1980-08-01

    The kinetics and morphology of sequestration and margination of rat leukocytes were studied using an isolated perfused and ventilated rat lung preparation. Whole rat blood, bone marrow suspension, or leukocyte suspensions, were used to perfuse the isolated rat lung. The lung was also perfused with latex particle suspensions and the passage of particles through the lung capillaries was studied. When a leukocyte suspension was perfused through the lung in the single-pass mode, the rate of sequestration decreased as more cells were perfused. In contrast, latex particles of a size comparable to that of leukocytes were totally stopped by the lung. When the leukocyte suspension was recirculated through the lung, cells were rapidly removed from circulation until a steady state was reached, after which no net removal of cells by the lung occurred. These results indicate that leukocytes are reversibly sequestered from circulation. The sequestered cells marginated and attached to the luminal surface of the endothelium of post-capillary venules and veins. A mathematical model was developed based on the assumption that the attachment and detachment of leukocytes to blood vessel walls follows first-order kinetics. The model correctly predicts the following characteristics of the system: (a) the kinetics of the sequestration of leukocytes by the lung; (b) the existence of a steady state when a suspension of leukocytes is recirculated through the lung; and (c) the independence of the fraction of cells remaining in circulation from the starting concentration for all values of starting concentration. (ERB)

  16. A study on trypsin, Aspergillus flavus and Bacillus sp. protease inhibitory activity in Cassia tora (L. syn Senna tora (L. Roxb. seed extract

    Directory of Open Access Journals (Sweden)

    Garg Satyendra K

    2011-07-01

    Full Text Available Abstract Background Proteases play an important role in virulence of many human, plant and insect pathogens. The proteinaceous protease inhibitors of plant origin have been reported widely from many plant species. The inhibitors may potentially be used for multiple therapeutic applications in viral, bacterial, fungal diseases and physiological disorders. In traditional Indian medicine system, Cassia tora (Senna tora is reportedly effective in treatment of skin and gastrointestinal disorders. The present study explores the protease inhibitory activity of the above plant seeds against trypsin, Aspergillus flavus and Bacillus sp. proteases. Methods The crushed seeds of Cassia tora were washed thoroughly with acetone and hexane for depigmentation and defatting. The proteins were fractionated by ammonium sulphate (0-30, 30-60, 60-90% followed by dialysis and size exclusion chromatography (SEC. The inhibitory potential of crude seed extract and most active dialyzed fraction against trypsin and proteases was established by spot test using unprocessed x-ray film and casein digestion methods, respectively. Electrophoretic analysis of most active fraction (30-60% and SEC elutes were carried employing Sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE and Gelatin SDS-PAGE. Inhibition of fungal spore germination was studied in the presence of dialyzed active inhibitor fraction. Standard deviation (SD and ANOVA were employed as statistical tools. Results The crude seeds' extract displayed strong antitryptic, bacterial and fungal protease inhibitory activity on x-ray film. The seed protein fraction 30-60% was found most active for trypsin inhibition in caseinolytic assay (P Aspergillus flavus and Bacillus sp. proteases remained only 4, 7 and 3.1%, respectively when proteases were incubated with 3 mg ml-1 seed protein extract for 60 min. The inhibitory activity was evident in gelatin SDS-PAGE where a major band (~17-19 kD of protease

  17. Leukocyte subsets and neutrophil function after short-term spaceflight

    Science.gov (United States)

    Stowe, R. P.; Sams, C. F.; Mehta, S. K.; Kaur, I.; Jones, M. L.; Feeback, D. L.; Pierson, D. L.

    1999-01-01

    Changes in leukocyte subpopulations and function after spaceflight have been observed but the mechanisms underlying these changes are not well defined. This study investigated the effects of short-term spaceflight (8-15 days) on circulating leukocyte subsets, stress hormones, immunoglobulin levels, and neutrophil function. At landing, a 1.5-fold increase in neutrophils was observed compared with preflight values; lymphocytes were slightly decreased, whereas the results were variable for monocytes. No significant changes were observed in plasma levels of immunoglobulins, cortisol, or adrenocorticotropic hormone. In contrast, urinary epinephrine, norepinephrine, and cortisol were significantly elevated at landing. Band neutrophils were observed in 9 of 16 astronauts. Neutrophil chemotactic assays showed a 10-fold decrease in the optimal dose response after landing. Neutrophil adhesion to endothelial cells was increased both before and after spaceflight. At landing, the expression of MAC-1 was significantly decreased while L-selectin was significantly increased. These functional alterations may be of clinical significance on long-duration space missions.

  18. Peripheral blood and milk leukocytes subsets of lactating Sarda ewes

    Directory of Open Access Journals (Sweden)

    Piero Bonelli

    2013-05-01

    Full Text Available Leukocytes subpopulations in blood and milk of lactating Sarda ewes were investigated. Animals characterized by a SSC level <500×103cells/mL and a negative bacteriological examination were sampled in early, mid and late lactation. Milk differential cell count evidenced that macrophage represented the main population (42.8%±3.5 followed by lymphocytes (40.2%±3.4 and neutrophils (8,6%±2.1. Flow cytometry analysis showed that lymphocytes subsets in milk were quite different from blood. High CD8+ and low CD4+ lymphocytes percentages determined a CD4/CD8 ratio inversion in milk compared to blood (0.3%±0.03 vs 1.8%±0.08. CD8+ decreased while, conversely, CD4+ increased in late lactation. γδ T cells were more represented in milk (12.6%±1.3 than in blood (6.8%±0.3 and their proportions appeared similar throughout lactation in both compartments. IL-2 receptor was mainly expressed in milk on T cytotoxic lymphocytes. Data obtained in uninfected mammary glands could allow an early discrimination between physiological and pathological changes occurring in ewe milk. Further phenotypical and functional studies on milk leukocytes subsets might help to understand defense mechanisms of the ovine mammary gland against IMI.

  19. Swine Leukocyte Antigen (SLA) Class II is a Xenoantigen.

    Science.gov (United States)

    Ladowski, Joseph M; Reyes, Luz M; Martens, Gregory R; Butler, James R; Wang, Zheng-Yu; Eckhoff, Devin E; Tector, Matt; Tector, A Joseph

    2017-08-24

    Over 130 000 patients in the United States alone need a life-saving organ transplant. Genetically modified porcine organs could resolve the donor organ shortage, but human xenoreactive antibodies destroy pig cells and are the major barrier to clinical application of xenotransplantation. The objective of this study was to determine whether waitlisted patients possess preformed antibodies to swine leukocyte antigen (SLA) class II, homologs of the class II human leukocyte antigens (HLA). Sera from people currently awaiting solid organ transplant were tested for IgG binding to class II SLA proteins when expressed on mammalian cells. Pig fibroblasts were made positive by transfection with the class II transactivator (CIITA). As a second expression system, transgenes encoding the alpha and beta chains of class II SLA were transfected into Human embryonic kidney (HEK293) cells. Human sera containing IgG specific for class II HLA molecules exhibited greater binding to class II SLA positive cells than to SLA negative cells. Sera lacking antibodies against class II HLA showed no change in binding regardless of the presence of class II SLA. These antibodies could recognize either SLA-DR or SLA-DQ complexes. Class II SLA proteins may behave as xenoantigens for people with humoral immunity towards class II HLA molecules.

  20. Global methylation of blood leukocyte DNA and risk of melanoma.

    Science.gov (United States)

    Shen, Jie; Song, Renduo; Wan, Jie; Huff, Chad; Fang, Shenying; Lee, Jeffrey E; Zhao, Hua

    2017-04-01

    Global DNA methylation, possibly influenced by lifestyle and environmental factors, has been suggested to play an active role in carcinogenesis. However, its role in melanoma has rarely been explored. The aims of this study were to evaluate the relationship between melanoma risk and levels of 5-methylcytosine (5-mC), a marker for global DNA methylation, in blood leukocyte DNA, and to determine whether this 5-mC level is influenced by pigmentation and sun exposure. This case-control study included 540 melanoma cases and 540 healthy controls. Overall, melanoma cases had significantly lower levels of 5-mC% than healthy controls (median: 3.24 vs. 3.91, p melanoma (OR: 1.25; 95% CI: 1.08, 1.37). A significant dose-response relationship was observed in quartile analysis (p = 0.001). Our results suggest that global hypomethylation in blood leukocyte DNA is associated with increased risk of melanoma and that the level of methylation is influenced by pigmentation and sun exposure. © 2016 UICC.

  1. Stable and unstable angina: Identifying novel markers on circulating leukocytes.

    Science.gov (United States)

    Brown, Angus; Lattimore, Jo-Dee; McGrady, Michele; Sullivan, David; Dyer, Wayne; Braet, Filip; Dos Remedios, Cristobal

    2008-01-01

    There is currently no blood-based test that can rapidly and objectively distinguish between chest pain which is initiated by increased myocardial oxygen demand (stable angina pectoris (SAP)) and chest pain initiated due to decreased coronary blood flow (unstable angina pectoris (UAP)). Since leukocytes play an active role in the progression of coronary artery disease (CAD), we hypothesize these can provide novel markers of SAP and UAP. Here we use a microarray of 82 cluster of differentiation (CD) antibodies (plus controls) to selectively immobilize peripheral blood mononuclear cells. We find that the pattern of leukocyte immobilization from patients with CAD significantly differs from healthy donors. Within the CAD group, 15 SAP patients exhibited significant (p<0.05) changes in 8 of 82 CD antibody spots compared to 19 age-matched healthy blood donors. An additional ten CD antigens differed between healthy donors and patients with UAP (p<0.05). Furthermore, seven CD antibody spots are significantly different between SAP and UAP patients. These preliminary data suggest it is now appropriate to undertake a larger clinical trial to test the hypothesis that these antibody microarrays can monitor the progression from SAP to UAP.

  2. Fcγ Receptor Heterogeneity in Leukocyte Functional Responses

    Science.gov (United States)

    Rosales, Carlos

    2017-01-01

    Antibodies participate in defense of the organism from all types of pathogens, including viruses, bacteria, fungi, and protozoa. IgG antibodies recognize their associated antigen via their two Fab portions and are in turn recognized though their Fc portion by specific Fcγ receptors (FcγRs) on the membrane of immune cells. Multiple types and polymorphic variants of FcγR exist. These receptors are expressed in many cells types and are also redundant in inducing cell responses. Crosslinking of FcγR on the surface of leukocytes activates several effector functions aimed toward the destruction of pathogens and the induction of an inflammatory response. In the past few years, new evidence on how the particular IgG subclass and the glycosylation pattern of the antibody modulate the IgG–FcγR interaction has been presented. Despite these advances, our knowledge of what particular effector function is activated in a certain cell and in response to a specific type of FcγR remains very limited today. On one hand, each immune cell could be programmed to perform a particular cell function after FcγR crosslinking. On the other, each FcγR could activate a particular signaling pathway leading to a unique cell response. In this review, I describe the main types of FcγRs and our current view of how particular FcγRs activate various signaling pathways to promote unique leukocyte functions. PMID:28373871

  3. Interactions between CD44 and hyaluronan in leukocyte trafficking

    Directory of Open Access Journals (Sweden)

    Braedon eMcDonald

    2015-02-01

    Full Text Available Recruitment of leukocytes from the bloodstream to inflamed tissues requires a carefully regulated cascade of binding interactions between adhesion molecules on leukocytes and endothelial cells. Adhesive interactions between CD44 and hyaluronan have been implicated in the regulation of immune cell trafficking within various tissues. In this review, the biology of CD44-hyaluronan interactions in cell trafficking is summarized, with special attention to neutrophil recruitment within the liver microcirculation. We describe the molecular mechanisms that regulate adhesion between neutrophil CD44 and endothelial hyaluronan, including recent evidence implicating serum-derived hyaluronan associated protein (SHAP as an important co-factor in the binding of hyaluronan to CD44 under flow conditions. CD44-hyaluronan mediated neutrophil recruitment has been shown to contribute to innate immune responses to invading microbes, as well as to the pathogenesis of many inflammatory diseases, including various liver pathologies. As a result, blockade of neutrophil recruitment by targeting CD44-HA interactions has proven beneficial as an anti-inflammatory treatment strategy in a number of animal models of inflammatory diseases.

  4. DNA integrity of human leukocytes after magnetic resonance imaging.

    Science.gov (United States)

    Szerencsi, Ágnes; Kubinyi, Györgyi; Váliczkó, Éva; Juhász, Péter; Rudas, Gábor; Mester, Ádám; Jánossy, Gábor; Bakos, József; Thuróczy, György

    2013-10-01

    This study focuses on the effects of high-field (3T) magnetic resonance imaging (MRI) scans on the DNA integrity of human leukocytes in vitro in order to validate the study where genotoxic effects were obtained and published by Lee et al. The scanning protocol and exposure situation were the same as those used under routine clinical brain MRI scan. Peripheral blood samples from healthy non-smoking male donors were exposed to electromagnetic fields (EMF) produced by 3T magnetic resonance imaging equipment for 0, 22, 45, 67, and 89 min during the scanning procedure. Samples of positive control were exposed to ionizing radiation (4 Gy of (60)Co-γ). Single breaks of DNA in leukocytes were detected by single-cell gel electrophoresis (Comet assay). Chromosome breakage, chromosome loss and micronuclei formations were detected by a micronucleus test (MN). Three independent experiments were performed. The data of comet tail DNA%, olive tail moment and micronucleus frequency showed no DNA damages due to MRI exposure. The results of the Comet assay and the micronucleus test indicate that the applied exposure of MRI does not appear to produce breaks in the DNA and has no significant effect on DNA integrity.

  5. Leukocyte responses to immobilized patterns of CXCL8.

    Science.gov (United States)

    Girrbach, Maria; Rink, Ina; Ladnorg, Tatjana; Azucena, Carlos; Heißler, Stefan; Haraszti, Tamás; Schepers, Ute; Schmitz, Katja

    2016-06-01

    The attachment of neutrophils to the endothelial surface and their migration towards the site of inflammation following chemokine gradients play an essential role in the innate immune response. Chemokines adhere to glycosaminoglycans on the endothelial surface to be detected by leukocytes and trigger their movement along surface- bound gradients in a process called haptotaxis. In assays to systematically study the response of leukocytes to surface-bound compounds both the spatial arrangement of the compound as well as the mode of immobilization need to be controlled. In this study microcontact printing was employed to create patterns of hydrophobic or functionalized thiols on gold-coated glass slides and CXCL8 was immobilized on the thiol coated areas using three different strategies. Human neutrophils adhered to the CXCL8-coated lines but not to the PEG-coated background. We could show that more cells adhered to CXCL8 adsorbed to hydrophobic octadecanethiol than on CXCL8 covalently bound to amino undecanethiol or CXCL8 specifically bound to immobilized heparin on aminothiol. Likewise general cell activity such as lamellipodia formation and random migration were most pronounced for CXCL8 adsorbed on a hydrophobic surface which may be attributed to the larger amounts of protein immobilized on this type of surface.

  6. Novel and known constituents from Buddleja species and their activity against leukocyte eicosanoid generation.

    Science.gov (United States)

    Liao, Y H; Houghton, P J; Hoult, J R

    1999-09-01

    We have undertaken a systematic survey of the genus Buddleja used in traditional Chinese medicine for antiinflammatory and other indications by testing extracts and isolated natural products for their activity against the enzymes of the arachidonate cascade. This was done by using elicited rat peritoneal leukocytes, a physiologically relevant established whole cell system that expresses both cyclo-oxygenase (COX) and 5-lipoxygenase (5-LOX) activity. Lipophilic extracts of B. globosa roots and B. myriantha stem exhibited inhibitory activities in the 5-LOX and COX enzyme assays, whereas those of B. officinalis flowers, B. yunanesis stems, and B. asiatica stems showed inhibitory activities only against COX. The phytochemical investigation of these extracts, and consequent structure elucidation of isolated compounds using spectroscopic data, led to the isolation from B. globosa of three new terpenoid compounds named dihydrobuddledin A, buddledone A, and buddledone B and four known compounds-buddledins A, B, and C and zerumbone; 12 known compounds from B. officinalis-calceolarioside, campneoside, verbascoside, echinacoside, forsythoside B, angoroside A, crocetin monogentibiosyl ester, acacetin, acacetin-7-O-alpha-L-rhamnopyranosyl (1-6)-beta-D-glucopyranoside, acacetin-7-O-alpha-L-rhamnopyranosyl (1-6)[alpha-L-rhamnopyranosyl (1-2)]-beta-D-glucopyranoside, songarosaponin A, delta-amyrone; and eight known compounds fromB. yunanesis-11,14-dihydroxy-8,11, 13-abietatrien-7-one, beta-sitosterol, verbascoside, echinacoside, forsythoside B, angoroside A, methylcatapol, and sucrose. Tests on the isolated compounds for inhibition of eicosanoid synthesis showed that buddledin A, crocetin monogentibiosyl ester, and acacetin exhibited an inhibitory effect on COX with IC(50) values of 13.7 microM, 28.2 microM, and 77.5 microM, respectively, whereas buddledin A exhibited inhibitory effect on 5-LOX with an IC(50) value of 50.4 microM.

  7. Production of fibrogenic cytokines by interleukin-2-treated peripheral blood leukocytes

    DEFF Research Database (Denmark)

    Kovacs, E J; Brock, B; Silber, I E;

    1993-01-01

    was tested for induction of fibroblast proliferation, collagen synthesis, and expression of cytokine genes. RESULTS: Supernatants from IL-2-treated peripheral blood leukocytes induced six times more fibroblast proliferation than medium from leukocytes cultured without IL-2. The expression of type I...

  8. Elevated leukocyte count is associated with periodontitis in Korean adults: the 2012-2014 KNHANES.

    Science.gov (United States)

    Kwon, Y-J; Jeon, K-J; Chung, T-H; Lee, Y-J

    2017-03-01

    Both an elevated leukocyte count and periodontitis share well-recognized associations with cardiometabolic diseases. This cross-sectional study aimed to identify whether the leukocyte count is associated with periodontitis in a nationally representative Korean adult population. Data from 9391 participants (3659 males and 5732 females) enrolled in 2012-2014 Korean National Health and Nutrition Examination Survey were analyzed. Leukocyte quartiles were categorized as follows: 3000 ≤ Q1 ≤ 4870, 4880 ≤ Q2 ≤5790, 5800 ≤ Q3 ≤ 6840, and 6850 ≤ Q4 ≤ 10000 cells/μl. Periodontitis was defined as scoring greater than or equal to 'code 3' in at least one site according to the WHO's Community Periodontal Index. The odds ratios (ORs) and 95% confidence intervals (95% CIs) for periodontitis in each leukocyte count quartile were calculated using multiple logistic regression analyses. The prevalence of periodontitis was directly correlated with increasing leukocyte quartiles: 19%, 20.4%, 24.3%, and 30.3%. Compared with the lowest leukocyte quartile group, the OR (95% CI) for periodontitis of the highest leukocyte quartile was 1.558 (1.285-1.891) after controlling for confounding factors. An elevated leukocyte count was positively associated with the presence of periodontitis. © 2016 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  9. Sex difference in leukocyte telomere length is ablated in opposite-sex co-twins

    NARCIS (Netherlands)

    Benetos, Athanase; Dalgard, Christine; Labat, Carlos; Kark, Jeremy D.; Verhulst, Simon; Christensen, Kaare; Kimura, Masayuki; Horvath, Kent; Kyvik, Kirsten Ohm; Aviv, Abraham

    2014-01-01

    Background: In eutherian mammals and in humans, the female fetus may be masculinized while sharing the intra-uterine environment with a male fetus. Telomere length (TL), as expressed in leukocytes, is heritable and is longer in women than in men. The main determinant of leukocyte TL (LTL) is LTL at

  10. Protein-bound uremic toxins stimulate crosstalk between leukocytes and vessel wall.

    Science.gov (United States)

    Pletinck, Anneleen; Glorieux, Griet; Schepers, Eva; Cohen, Gerald; Gondouin, Bertrand; Van Landschoot, Maria; Eloot, Sunny; Rops, Angelique; Van de Voorde, Johan; De Vriese, An; van der Vlag, Johan; Brunet, Philippe; Van Biesen, Wim; Vanholder, Raymond

    2013-12-01

    Leukocyte activation and endothelial damage both contribute to cardiovascular disease, a major cause of morbidity and mortality in CKD. Experimental in vitro data link several protein-bound uremic retention solutes to the modulation of inflammatory stimuli, including endothelium and leukocyte responses and cardiovascular damage, corroborating observational in vivo data. However, the impact of these uremic toxins on the crosstalk between endothelium and leukocytes has not been assessed. This study evaluated the effects of acute and continuous exposure to uremic levels of indoxylsulfate (IS), p-cresylsulfate (pCS), and p-cresylglucuronide (pCG) on the recruitment of circulating leukocytes in the rat peritoneal vascular bed using intravital microscopy. Superfusion with IS induced strong leukocyte adhesion, enhanced extravasation, and interrupted blood flow, whereas pCS caused a rapid increase in leukocyte rolling. Superfusion with pCS and pCG combined caused impaired blood flow and vascular leakage but did not further enhance leukocyte rolling over pCS alone. Intravenous infusion with IS confirmed the superfusion results and caused shedding of heparan sulfate, pointing to disruption of the glycocalyx as the mechanism likely mediating IS-induced flow stagnation. These results provide the first clear in vivo evidence that IS, pCS, and pCG exert proinflammatory effects that contribute to vascular damage by stimulating crosstalk between leukocytes and vessels.

  11. Broadly altered gene expression in blood leukocytes in essential hypertension is absent during treatment

    NARCIS (Netherlands)

    Chon, H; Gaillard, CAJM; van der Meijden, BB; Dijstelbloem, HM; Kraaijenhagen, RJ; van Leenen, D; Holstege, FCP; Joles, JA; Bluyssen, HAR; Koomans, HA; Braam, B

    2004-01-01

    We assessed whether large-scale expression profiling of leukocytes of patients with essential hypertension reflects characteristics of systemic disease and whether such changes are responsive to antihypertensive therapy. Total RNA from leukocytes were obtained from untreated (n=6) and treated (n=6)

  12. Blockade of leukocyte haptokinesis and haptotaxis by ketoprofen, diclofenac and SC-560

    Directory of Open Access Journals (Sweden)

    Paskauskas Saulius

    2011-11-01

    Full Text Available Abstract Background Nonsteroidal anti-inflammatory drugs (NSAID represent a one of the most widely used anti-inflammatory substances. Their anti-inflammatory effects are mainly based on inhibition of cyclooxygenase. The potential direct effect of NSAID on leukocyte migration was poorly investigated. Using time-lapse microscopy and 96-well fluorescence-based assay, we studied the effect of three different NSAID, ketoprofen, diclofenac and SC-560, on leukocyte haptokinesis and haptotaxis in vivo and in vitro. Results NSAID induced an immediate inhibiting effect on leukocyte migration both in vitro and in vivo. This effect was dose-dependent and was not restricted to a specific type of leukocytes. The inhibition of leukocyte migration by NSAID was partially re-stored after removal of inhibiting agent. Only complete blockade of leukocyte migration was accompanied by a strong reduction of [Ca2+]i. Conclusions NSAID strongly supress leukocyte migration. The results of the present study may have important clinical implications since blockade of leukocyte migration can be achieved after topical application of NSAID.

  13. Leukocyte accumulation promoting fibrin deposition is mediated in vivo by P-selectin on adherent platelets

    Science.gov (United States)

    Palabrica, Theresa; Lobb, Roy; Furie, Barbara C.; Aronovitz, Mark; Benjamin, Christopher; Hsu, Yen-Ming; Sajer, Susan A.; Furie, Bruce

    1992-10-01

    THE glycoprotein P-selectin is a cell adhesion molecule of stimulated platelets and endothelial cells, which mediates the interaction of these cells with neutrophils and monocytes1,2. It is a membrane component of cell storage granules3-6, and is a member of the selectin family which includes E-selectin and L-selectin7,8. P-selectin recognizes both lineage-specific carbohydrate ligands on monocytes and neutrophils, including the Lewis x antigen, sialic acid, and a protein component9-12. In inflammation and thrombosis, P-selectin may mediate the interaction of leukocytes with platelets bound in the region of tissue injury and with stimulated endothelium1,2. To evaluate the role of P-selectin in platelet-leukocyte adhesion in vivo, the accumulation of leukocytes within an experimental thrombus was explored in an arteriovenous shunt model in baboons13. A Dacron graft implanted within an arteriovenous shunt is thrombogenic, accumulating platelets and fibrin within its lumen. These bound platelets express P-selectin14. Here we show that antibody inhibition of leukocyte binding to P-selectin expressed on platelets immobilized on the graft blocks leukocyte accumulation and inhibits the deposition of fibrin within the thrombus. These results indicate that P-selectin is an important adhesion molecule on platelets, mediating platelet-leukocyte binding in vivo, that the presence of leukocytes in thrombi is mediated by P-selectin, and that these leukocytes promote fibrin deposition.

  14. Tc-99m HM-PAO labelling of leukocytes for detection of inflammatory bowel disease

    Energy Technology Data Exchange (ETDEWEB)

    Schuemichen, C.; Schoelmerich, J.

    1986-10-01

    Labelling of purified leukocytes with the brain imaging agent Tc-99m HM-PAO proved to be a reliable method, yielding a 45% labelling efficiency in phosphate buffer and leaving more than 85% of the leukocytes viable. In fifteen patients with M. Crohn, a positive finding was shown in all cases on the six hour image.

  15. Signaling through MyD88 regulates leukocyte recruitment after brain injury

    DEFF Research Database (Denmark)

    Babcock, Alicia A; Toft-Hansen, Henrik; Owens, Trevor

    2008-01-01

    Injury to the CNS provokes an innate inflammatory reaction that engages infiltrating leukocytes with the capacity to repair and/or exacerbate tissue damage. The initial cues that orchestrate leukocyte entry remain poorly defined. We have used flow cytometry to investigate whether MyD88, an adapto...

  16. Sex difference in leukocyte telomere length is ablated in opposite-sex co-twins

    DEFF Research Database (Denmark)

    Benetos, Athanase; Dalgård, Christine; Labat, Carlos;

    2014-01-01

    BACKGROUND: In eutherian mammals and in humans, the female fetus may be masculinized while sharing the intra-uterine environment with a male fetus. Telomere length (TL), as expressed in leukocytes, is heritable and is longer in women than in men. The main determinant of leukocyte TL (LTL) is LTL...

  17. ICAM-1 enrichment near tri-cellular endothelial junctions is preferentially associated with leukocyte transmigration, and signals for reorganization of these junctions to accommodate leukocyte passage

    Science.gov (United States)

    Sumagin, Ronen; Sarelius, Ingrid H

    2010-01-01

    Leukocyte transmigration occurs at specific locations (portals) on the endothelium, but the nature of these portals is not clear. Using intravital confocal microscopy of anesthetized mouse cremaster muscle in combination with immunofluorescence labeling, we showed that in microvessels transmigration is mainly junctional and preferentially occurs at tri-cellular endothelial junctional regions. Our data suggest that enrichment of ICAM-1 near approximately 43% of these junctions makes these locations preferred for transmigration, by signaling the location of a nearby portal, as well as preparing the EC-junctions to accommodate leukocyte passage. Blockade of the extracellular domain of the ICAM-1 significantly reduced transmigration (by 68.8±4.5%), by reducing the ability of leukocytes to get to these portals. In contrast, blockade of the cytoplasmic tail of ICAM-1 reduced transmigration (by 71.1±7.0%) by disabling VE-Cadherin rearrangement. Importantly, venular convergences are optimally equipped to support leukocyte transmigration. Differences in EC morphology result in a significantly higher number of tri-cellular junctions in convergences compared to straight venular regions (20.7±1.2 vs 12.43±1.1/6000μm2, respectively). Consequently leukocyte adhesion and transmigration are significantly higher in convergences compared to straight regions (1.6- and 2.6-fold, respectively). Together, these data identify an important role for EC morphology and expression patterns of ICAM-1 in leukocyte transmigration. PMID:20363969

  18. Characteristics of an autologous leukocyte and platelet-rich fibrin patch intended for the treatment of recalcitrant wounds.

    Science.gov (United States)

    Lundquist, Rasmus; Holmstrøm, Kim; Clausen, Christian; Jørgensen, Bo; Karlsmark, Tonny

    2013-01-01

    We have investigated the physical, biochemical, and cellular properties of an autologous leukocyte and platelet-rich fibrin patch. This was generated in an automated device from a sample of a patient's blood at the point of care. Using microscopy, cell counting, enzyme-linked immunosorbent assay, antibody arrays, and cell culture assays, we show that the patch is a three-layered membrane comprising a fibrin sheet, a layer of platelets, and a layer of leukocytes. Mean recovery of platelets from the donated blood was 98% (±95%CI 0.8%). Mean levels of platelet-derived growth factor AB, human transforming growth factor beta 1, and vascular endothelial growth factor extracted from the patch were determined as 127 ng (±95% CI 20), 92 ng (±95%CI 17), and 1.35 ng (±95%CI 0.37), respectively. We showed a continued release of PDGF-AB over several days, the rate of which was increased by the addition of chronic wound fluid. By comparison with traditional platelet-rich plasma, differences in immune components were found. The relevance of these findings was assessed by showing a mitogenic and migratory effect on cultured human dermal fibroblasts. Further, we showed that fibrocytes, a cell type important for acute wound healing, could be grown from the patch. The relevance of these findings in relation to the use of the patch for treating recalcitrant wounds is discussed.

  19. Morinda citrifolia linn leaf extract possesses antioxidant activities and reduces nociceptive behavior and leukocyte migration

    OpenAIRE

    Serafini, Mairim Russo; Santos, Rodrigo Correia; Guimarães, Adriana Gibara; Santos, João Paulo Almeida dos; Santos, Alan Diego da Conceição; Alves, Izabel Almeida; Gelain, Daniel Pens; Nogueira, Paulo Cesar de Lima; Quintans-Júnior, Lucindo José; Bonjardim, Leonardo Rigoldi; Araújo, Adriano Antunes de Souza

    2011-01-01

    This is a copy of an article published in the Journal of Medicinal Food© 2011 - copyright Mary Ann Liebert, Inc.; Journal of Medicinal Food is available online at: http://online.liebertpub.com Herbal drugs have been used since ancient times to treat a wide range of diseases. Morinda citrifolia Linn (popularly known as ‘‘Noni’’) has been used in folk medicine by Polynesians for over 2,000 years. It is reported to have a broad range of therapeutic effects, including effects against headache,...

  20. In-111-labeled leukocyte brain SPECT imaging. Clinical significance in evaluating acute ischemic stroke

    Energy Technology Data Exchange (ETDEWEB)

    Fujinuma, Kunihiko [Kitasato Univ., Sagamihara, Kanagawa (Japan). School of Medicine

    2002-02-01

    Many experimental studies have demonstrated that leukocyte infiltration plays an important role in the progression of ischemic cellular damage or post perfusion brain injury. However, only a few clinical studies have been reported. The purpose of this study is to evaluate the clinical significance of leukocyte accumulation in the ischemic brain tissue. Seventy six patients (49 men, 27 women; mean age: 65.5{+-}13.9 years) with acute ischemic stroke were studied by leukocyte brain SPECT imaging. A diagnosis included cardioembolism (n=46), atherothrombotic infarction (n=24), TIA (n=3) and lacuna (n=3). Immediately after the CBF study using Tc-99m-ECD (600 MBq), indium-111-labeled autologous leukocytes were injected. A brain scan for leukocytes was performed 48 hours later. The leukocyte-SPECT study was made 11.1{+-}7.7 days after the onset of stroke. Regional accumulation of leukocytes in the ischemic tissue was evaluated both by visual assessment and by measuring the hemispheric asymmetry index for leukocyte (AI-leuko), and was evaluated by comparison with variable factors including age, gender, infarction size, hemorrhagic transformation, timing of study after the onset, type of stroke and functional outcome. Of the 61 patients with acute ischemic stroke within 2 weeks of onset, 28 patients showed the accumulation of leukocytes in the central zone of ischemia. Six of 7 patients with repeated studies showed a reduction in leukocyte accumulation with time after the onset. Factors significantly associated with the higher accumulation of leukocyte included cardioembolic stroke, larger size of infarct, presence of hemorrhagic transformation and significant reduction in flow. In the 61 patients within 2 weeks of onset, the functional outcome was significantly correlated with the accumulation of leukocyte (p<0.001). The accumulation of leukocytes was seen more in patients with embolic stroke, larger infarction, and hemorrhagic transformation. The higher accumulation

  1. Preclinical studies for the gene therapy of leukocyte adhesion deficiency type I

    Energy Technology Data Exchange (ETDEWEB)

    Leon Rico, D.

    2015-07-01

    Leukocyte Adhesion Deficiency Type I (LAD-I) is a primary immunodeficiency caused by mutations in ITGB2 gene, encoding for CD18 protein (also known as 2 subunit). This protein binds to different CD11 subunits to form 2 integrins, which are expressed in the leukocyte membrane and allow leukocytes to firmly adhere to the endothelium as a previous step to the extravasation. In LAD-I patients, ITGB2 mutations lead to absent, low or aberrant CD18 expression, which results in absent or low 2 integrin expression on the leukocyte membrane. CD18 deficient leukocytes, especially neutrophils, fail to extravasate from the bloodstream to infected tissues. LAD-I patients suffer from recurrent and severe infections leading normally to death. (Author)

  2. Rosmarinus officinalis L. essential oil inhibits in vivo and in vitro leukocyte migration.

    Science.gov (United States)

    Nogueira de Melo, Gessilda Alcantara; Grespan, Renata; Fonseca, Jefferson Pitelli; Farinha, Thiago Oliveira; Silva, Expedito Leite; Romero, Adriano Lopes; Bersani-Amado, Ciomar A; Cuman, Roberto Kenji Nakamura

    2011-09-01

    Rosmarinus officinalis L. (Lamiaceae), popularly known as rosemary, is used for food flavoring and in folk medicine as an antispasmodic, analgesic, antirheumatic, diuretic, and antiepileptic agent. Few studies have shown the anti-inflammatory effects of rosemary essential oil (REO). This study evaluated the effects of REO on leukocyte migration through in vivo leukocyte migration and in vitro chemotaxis assay. REO was analyzed by using gas chromatography-mass spectometry, and the main components identified were camphor (27.59%), 1,8-cineole (15.74%), α-pinene (16.58%), and β-myrcene (10.02%). In rats, administration of REO reduced the number of leukocytes that rolled, adhered, and migrated to the scrotal chamber after carrageenan injection. All doses of REO tested significantly inhibited leukocyte chemotaxis induced by casein. The effects of REO on leukocyte migration highlight an important mechanism of the anti-inflammatory action of rosemary.

  3. Influence of antioxidant complex on the adhesion of leukocytes in chronic venous insufficiency of lower limbs in rats

    Directory of Open Access Journals (Sweden)

    Mark Plotnikov

    2012-01-01

    Conclusions: Model of CVI of lower limb is accompanied by increased venous pressure and raised adhesion activity of leukocytes. Administration of AOC for 14 days reduces the adhesive activity of leukocytes.

  4. Filtration of activated granulocytes during cardiopulmonary bypass surgery : A morphologic and immunologic study to characterize the trapped leukocytes

    NARCIS (Netherlands)

    Smit, JJJ; de Vries, AJ; Gu, YJ; van Oeveren, W

    Cardiopulmonary bypass surgery induces an inflammatory reaction among others by activation of granulocytes. Leukocyte filtration has been shown to reduce the postoperative morbidity mediated by activated granulocytes. However, little is known about the mechanism of filter-leukocyte interaction, This

  5. Filtration of activated granulocytes during cardiopulmonary bypass surgery : A morphologic and immunologic study to characterize the trapped leukocytes

    NARCIS (Netherlands)

    Smit, JJJ; de Vries, AJ; Gu, YJ; van Oeveren, W

    2000-01-01

    Cardiopulmonary bypass surgery induces an inflammatory reaction among others by activation of granulocytes. Leukocyte filtration has been shown to reduce the postoperative morbidity mediated by activated granulocytes. However, little is known about the mechanism of filter-leukocyte interaction, This

  6. Leukotriene C4 biosynthesis in isolated August rat peritoneal leukocytes

    Directory of Open Access Journals (Sweden)

    J. M. Huebner

    1996-01-01

    Full Text Available The mixed leukocyte population obtained from the peritoneum of the August rat is a potentially important experimental model of inherent eosinophilia that has not been well characterized. In the present study, isolated cell preparations generated a concentration-dependent release of leukotriene (LT C4 when exposed to the Ca2+ ionophore A23187, reaching maximal stimulation at 5.0 μM. This response was inhibited by the 5-lipoxygenase activating protein antagonist MK-886 (0.1 μM, nominally Ca2+ and Mg2+-free incubation media and by activation of protein kinase C via phorbol 12-myristate 13-acetate (50 nM. These findings establish a model system for investigating LTC4 profiles contingent with innate peritoneal eosinophilia and are consistent with the hypothesis that cellular LTC4 biosynthesis is phosphoregulated.

  7. Characterization of Leukocyte-platelet Rich Fibrin, A Novel Biomaterial.

    Science.gov (United States)

    Madurantakam, Parthasarathy; Yoganarasimha, Suyog; Hasan, Fadi K

    2015-09-29

    Autologous platelet concentrates represent promising innovative tools in the field of regenerative medicine and have been extensively used in oral surgery. Unlike platelet rich plasma (PRP) that is a gel or a suspension, Leukocyte-Platelet Rich Fibrin (L-PRF) is a solid 3D fibrin membrane generated chair-side from whole blood containing no anti-coagulant. The membrane has a dense three dimensional fibrin matrix with enriched platelets and abundant growth factors. L-PRF is a popular adjunct in surgeries because of its superior handling characteristics as well as its suturability to the wound bed. The goal of the study is to demonstrate generation as well as provide detailed characterization of relevant properties of L-PRF that underlie its clinical success.

  8. Diterpenoids from Tetraclinis articulata that inhibit various human leukocyte functions.

    Science.gov (United States)

    Barrero, Alejandro F; Quílez del Moral, José F; Lucas, Rut; Payá, Miguel; Akssira, Mohamed; Akaad, Said; Mellouki, Fouad

    2003-06-01

    Ten new compounds, eight of them pimarane derivatives (1-8), together with a menthane dimer (9) and a totarane diterpenoid (10), were isolated from the leaves and wood of Tetraclinis articulata. The structures of 1-10 were established by using spectroscopic techniques, including 2D NMR spectra. Pimaranes 1-5 were found to possess an unusual cis interannular union of the B and C rings, which, from a biogenetic perspective, could be derived from the hydration of a carbocation at C-8. Compounds 4-6 and a mixture of 7 and 11 modulated different human leukocyte functions at a concentration of 10 microM, mainly the degranulation process measured as myeloperoxidase release and, to a lesser extent, the superoxide production measured by chemiluminescence.

  9. In vitro effect of benzodiazepines on polymorphonuclear leukocyte oxidative activity.

    Science.gov (United States)

    Goldfarb, G; Belghiti, J; Gautero, H; Boivin, P

    1984-01-01

    The effect of three benzodiazepine compounds, diazepam, flunitrazepam, and clorazepate, on oxidative activity of human polymorphonuclear leukocyte (PMN) was investigated. The oxidative activity of zymosan-stimulated PMN in the presence of three concentrations (10, 20, and 40 micrograms/ml) of these compounds was measured polarographically. In addition, zymosan-stimulated nitroblue tetrazolium (NBT) reduction was measured in the presence of various concentrations of flunitrazepam. All three compounds inhibited oxygen consumption of the PMN. The extent of inhibition was linear with respect to log-concentrations; oxygen consumption was reduced 50% for concentrations of diazepam, flunitrazepam, and clorazepate of 13 micrograms/ml, 56 micrograms/ml, and 285 micrograms/ml, respectively. In addition 30% and 100% inhibition of NBT reduction by flunitrazepam were observed at respective concentrations of 10 micrograms/ml and 60 micrograms/ml. The clinical relevance of these findings remains to be determined.

  10. Leukocyte apheresis in the management of ulcerative colitis

    Directory of Open Access Journals (Sweden)

    Helmy Ahmed

    2009-01-01

    Full Text Available Ulcerative colitis is a chronic inflammatory disease that affects the colon and rectum. Its pathogenesis is probably multifactorial including the influx of certain cytokines into the colonic mucosa, causing disease activity and relapse. The hypothesis of removing such cytokines from the circulation by leukocytapheresis was implemented to reduce disease activity, maintain remission, and prevent relapse. Many recent reports not only in Japan, but also in the West, have highlighted its beneficial effects in both adult and pediatric patients. Large placebo-controlled studies are needed to confirm the available data in this regard. In this article, we shed some light on the use of leukocyte apheresis in the management of autoimmune diseases, especially ulcerative colitis.

  11. Human Leukocyte Antigen Diversity: A Southern African Perspective

    Directory of Open Access Journals (Sweden)

    Mqondisi Tshabalala

    2015-01-01

    Full Text Available Despite the increasingly well-documented evidence of high genetic, ethnic, and linguistic diversity amongst African populations, there is limited data on human leukocyte antigen (HLA diversity in these populations. HLA is part of the host defense mechanism mediated through antigen presentation to effector cells of the immune system. With the high disease burden in southern Africa, HLA diversity data is increasingly important in the design of population-specific vaccines and the improvement of transplantation therapeutic interventions. This review highlights the paucity of HLA diversity data amongst southern African populations and defines a need for information of this kind. This information will support disease association studies, provide guidance in vaccine design, and improve transplantation outcomes.

  12. Susceptibility of different leukocyte cell types to Vaccinia virus infection

    Directory of Open Access Journals (Sweden)

    Sánchez-Puig Juana M

    2004-11-01

    Full Text Available Abstract Background Vaccinia virus, the prototype member of the family Poxviridae, was used extensively in the past as the Smallpox vaccine, and is currently considered as a candidate vector for new recombinant vaccines. Vaccinia virus has a wide host range, and is known to infect cultures of a variety of cell lines of mammalian origin. However, little is known about the virus tropism in human leukocyte populations. We report here that various cell types within leukocyte populations have widely different susceptibility to infection with vaccinia virus. Results We have investigated the ability of vaccinia virus to infect human PBLs by using virus recombinants expressing green fluorescent protein (GFP, and monoclonal antibodies specific for PBL subpopulations. Flow cytometry allowed the identification of infected cells within the PBL mixture 1–5 hours after infection. Antibody labeling revealed that different cell populations had very different infection rates. Monocytes showed the highest percentage of infected cells, followed by B lymphocytes and NK cells. In contrast to those cell types, the rate of infection of T lymphocytes was low. Comparison of vaccinia virus strains WR and MVA showed that both strains infected efficiently the monocyte population, although producing different expression levels. Our results suggest that MVA was less efficient than WR in infecting NK cells and B lymphocytes. Overall, both WR and MVA consistently showed a strong preference for the infection of non-T cells. Conclusions When infecting fresh human PBL preparations, vaccinia virus showed a strong bias towards the infection of monocytes, followed by B lymphocytes and NK cells. In contrast, very poor infection of T lymphocytes was detected. These finding may have important implications both in our understanding of poxvirus pathogenesis and in the development of improved smallpox vaccines.

  13. Detection of CFTR protein in human leukocytes by flow cytometry.

    Science.gov (United States)

    Johansson, Jan; Vezzalini, Marzia; Verzè, Genny; Caldrer, Sara; Bolognin, Silvia; Buffelli, Mario; Bellisola, Giuseppe; Tridello, Gloria; Assael, Baroukh Maurice; Melotti, Paola; Sorio, Claudio

    2014-07-01

    Leukocytes have previously been shown to express detectable levels of the protein cystic fibrosis transmembrane conductance regulator (CFTR). This study aims to evaluate the application of flow cytometric (FC) analysis to detect CFTR expression, and changes thereof, in these cells. Aliquots (200 μL) of peripheral whole blood from 12 healthy control volunteers (CTRLs), 12 carriers of a CFTR mutation (CFC), and 40 patients with cystic fibrosis (CF) carrying various combinations of CFTR mutations were incubated with specific fluorescent probes recognizing CFTR protein expressed on the plasma membrane of leukocytes. FC was applied to analyze CFTR expression in monocytes, lymphocytes, and polymorphonuclear (PMN) cells. CFTR protein was detected in monocytes and lymphocytes, whereas inconclusive results were obtained from the analysis of PMN cells. Mean fluorescence intensity (MFI) ratio value and %CFTR-positive cells above a selected threshold were the two parameters selected to quantify CFTR expression in cells. Lowest variability and the highest reproducibility were obtained when analyzing monocytes. ANOVA results indicated that both parameters were able to discriminate monocytes of healthy controls and CF individuals according to CFTR mutation classes with high accuracy. Significantly increased MFI ratio values were recorded in CFTR-defective cells that were also able to improve CFTR function after ex vivo treatment with PTC124 (Ataluren), an investigative drug designed to permit the ribosome to read through nonsense CFTR mutations. The method described is minimally invasive and may be used in the monitoring of responses to drugs whose efficacy can depend on increased CFTR protein expression levels. © 2014 International Society for Advancement of Cytometry.

  14. Relationship between leukocyte count and angiographical characteristics of coronary atherosclerosis

    Institute of Scientific and Technical Information of China (English)

    En-zhi JIA; Zhi-jian YANG; Biao YUAN; Xiao-ling ZANG; Rong-hu WANG; Tie-bing ZHU; Lian-sheng WANG; Bo CHEN; Wen-zhu MA

    2005-01-01

    Aim: To explore the relationship between differential leucocyte count and coronary atherosclerosis. Methods: The study population consisted of 507 consecutive patients (376 male and 131 female) who underwent coronary angiography for suspected or known coronary atherosclerosis. The patients' smoking and drinking habits were investigated, and anthropometric measurements, serum measurements, and hematological measurements were conducted for every patient.The severity of coronary atherosclerosis was defined by using Gensini' s score system. One-way ANOVA, Spearman's correlation analysis, and multivariate stepwise linear regression analysis were employed to explore the relationship between differential leucocyte count and coronary atherosclerosis. Results: Oneway ANOVA indicated that the diastolic blood pressure, glucose, urea, creatinine,leukocyte count, neutrophil count, monocyte count, hemoglobin, and platelet count differed among the groups according to Gensini's score, the tertile values of which were used as cutoff points. Spearman's correlation analysis suggested that Gensini's score was significantly correlated with age, diastolic blood pressure,glucose, urea, creatinine, leukocyte count, neutrophil count, monocyte count,hemoglobin, and erythrocyte count, respectively. Multivariate stepwise linear regression analysis show that neutrophil count (β=0.247, P=0.000), age (β=0.141,P=0.001), glucose (β=0.173, P=0.000), creatinine (β=0.088, P=0.063), hemoglobin (β=-0.168, P=0.013) and sex (men were coded as 1 and women were coded as 2;β=-0.121, P=0.012) were significantly independently associated with the Gensini's score. Conclusion: The independent association of neutrophil count with the angiographical characteristics of coronary atherosclerosis, as estimated by Gensini's score, strongly suggests that granulocytosis may play a role in the development of coronary atherosclerosis.

  15. Using milk leukocyte differentials for diagnosis of subclinical bovine mastitis.

    Science.gov (United States)

    Gonçalves, Juliano Leonel; Lyman, Roberta L; Hockett, Mitchell; Rodriguez, Rudy; Dos Santos, Marcos Veiga; Anderson, Kevin L

    2017-08-01

    This research study aimed to evaluate the use of the milk leukocyte differential (MLD) to: (a) identify quarter milks that are culture-positive; and (b) characterize the milk leukocyte responses to specific groups of pathogens causing subclinical mastitis. The MLD measures the absolute number and relative percentage of inflammatory cells in milk samples. Using the MLD in two dairy herds (170 and 172 lactating cows, respectively), we studied all lactating cows with a most recent monthly Dairy Herd Improvement Association somatic cell count (SCC) >200 × 103 cells/ml. Quarter milk samples from 78 cows meeting study criteria were analysed by MLD and aseptically collected milk samples were subjected to microbiological culture (MC). Based upon automated instrument evaluation of the number and percentage of inflammatory cells in milk, samples were designated as either MLD-positive or - negative for subclinicial mastitis. Positive MC were obtained from 102/156 (65·4%) of MLD-positive milk samples, and 28/135 (20·7%) of MLD-negative milk samples were MC-positive. When MC was considered the gold standard for mastitis diagnosis, the calculated diagnostic Se of the MLD was 65·4% (IC95% = 57·4 to 72·8%) and the Sp was 79·3% (IC95% = 71·4 to 85·7%). Quarter milks positive on MC had higher absolute numbers of neutrophils, lymphocytes and macrophages, with higher neutrophils% and lymphocytes% but lower macrophages%. The Log10 (N/L) ratios were the most useful ratio to differentiate specific subclinical mastitis quarters from healthy quarters. Use of the MLD on cows with monthly composite SCC > 200 × 103 cells/ml for screening at quarter level identified quarters more likely to be culture-positive. In conclusion, the MLD can provide an analysis of mammary quarter status more detailed than provided by SCC alone; however, the MLD response to subclinical mastitis was not found useful to specifically identify the causative pathogen.

  16. Human Leukocyte Antigen-G and Regulatory T Cells during Specific Immunotherapy for Pollen Allergy

    DEFF Research Database (Denmark)

    Sørensen, Anja Elaine; Johnsen, Claus R; Dalgaard, Louise Torp;

    2013-01-01

    with pollen extract in vitro and immune factors were evaluated. Results: During SIT, the main changes in the peripheral blood were an increase in CXCR3+CD4+CD25+CD127low/- Tregs and a decrease in CCR4+CD4+CD25+CD127low/- Tregs, an increase in allergen-specific IgG4, and a decrease in sHLA-G during the first......Background: TH2-biased immune responses are important in allergy pathogenesis. Mechanisms of allergen-specific immunotherapy (SIT) might include the induction of regulatory T cells (Tregs) and immunoglobulin (Ig) G4 blocking antibodies, a reduction in the number of effector cells, and skewing...... of the cytokine profile towards a TH1-polarized immune response. We investigated the effects of SIT on T cells, on immunomodulation of human leukocyte antigen (HLA)-G, which has been associated with allergy, on regulatory cytokine expression, and on serum allergen-specific antibody subclasses (IgE and IgG4...

  17. Leukocyte telomere length and mortality among U.S. adults: Effect modification by physical activity behaviour.

    Science.gov (United States)

    Loprinzi, Paul D; Loenneke, Jeremy P

    2017-02-17

    The purpose of this study was to examine the association between leukocyte telomere length (LTL) and mortality (outcome variable), with consideration by physical activity behaviour. Data from the 1999-2002 National Health and Nutrition Examination Survey were employed (N = 6,611; 20-85 yrs), with follow-up mortality assessment through 31 December 2006. DNA was extracted from whole blood to assess LTL via quantitative polymerase chain reaction. Compared to those in the first LTL tertile, the adjusted hazard ratio for all-cause mortality for those in the 2(nd) and 3(rd) LTL tertiles, respectively, was 0.82 (95% CI: 0.60-1.12; P = .22) and 0.76 (95% CI: 0.50-1.14; P = .18). However, after adjustments, LTL tertile 3 (vs. 1) was associated with all-cause mortality (HR = 0.37; 95% CI: 0.14-0.93; P = .03) for those who engaged in moderate-intensity exercise. Similarly, LTL was associated with CVD-specific mortality for those who engaged in moderate-intensity exercise (HR = 0.17; 95% CI: 0.04-0.73; P = .02). Longer telomeres are associated with increased survival, particularly among men and those who are active, underscoring the importance of promotion of physical activity behaviour.

  18. Leukocyte transcript alterations in West-African girls following a booster vaccination with diphtheria-tetanus-pertussis vaccine

    DEFF Research Database (Denmark)

    Orntoft, Nikolaj W; Thorsen, Kasper; Benn, Christine S;

    2013-01-01

    Background. Observational studies from low-income countries have shown that the vaccination against diphtheria, tetanus and pertussis (DTP) is associated with excess female mortality due to infectious diseases. Methods. To investigate possible changes in gene expression after DTP vaccination, we...... identified a group of nine comparable West African girls, from a biobank of 356 children, who were due to receive DTP booster vaccine at age 18 months. As a pilot experiment we extracted RNA from blood samples before, and 6 weeks after, vaccination to analyze the coding transcriptome in leukocytes using...... expression microarrays, and ended up with information from eight girls. The data was further analyzed using dedicated array pathway and network software. We aimed to study whether DTP vaccination introduced a systematic alteration in the immune system in girls. Results. We found very few transcripts to alter...

  19. Tapasin discriminates peptide-human leukocyte antigen-A*02:01 complexes formed with natural ligands

    DEFF Research Database (Denmark)

    Røder, Gustav Andreas; Geironson, Linda; Rasmussen, Michael

    2011-01-01

    A plethora of peptides are generated intracellularly, and most peptide-human leukocyte antigen (HLA)-I interactions are of a transient, unproductive nature. Without a quality control mechanism, the HLA-I system would be stressed by futile attempts to present peptides not sufficient for the stable...... according to the identity of the peptide. The facilitation was also specific for the identity of the HLA-I heavy chain, where it correlated to established tapasin dependence hierarchies. Two large sets of HLA-A*02:01 binding peptides, one extracted from natural HLA-I ligands from the SYFPEITHI database...... functionally discriminate the selected SYFPEITHI peptides from the other peptide binders with high sensitivity and specificity. We suggest that this HLA-I- and peptide-specific function, together with the functions exerted by the more C-terminal parts of tapasin, are major features of tapasin-mediated HLA...

  20. A high-throughput microfluidic approach for 1000-fold leukocyte reduction of platelet-rich plasma

    Science.gov (United States)

    Xia, Hui; Strachan, Briony C.; Gifford, Sean C.; Shevkoplyas, Sergey S.

    2016-10-01

    Leukocyte reduction of donated blood products substantially reduces the risk of a number of transfusion-related complications. Current ‘leukoreduction’ filters operate by trapping leukocytes within specialized filtration material, while allowing desired blood components to pass through. However, the continuous release of inflammatory cytokines from the retained leukocytes, as well as the potential for platelet activation and clogging, are significant drawbacks of conventional ‘dead end’ filtration. To address these limitations, here we demonstrate our newly-developed ‘controlled incremental filtration’ (CIF) approach to perform high-throughput microfluidic removal of leukocytes from platelet-rich plasma (PRP) in a continuous flow regime. Leukocytes are separated from platelets within the PRP by progressively syphoning clarified PRP away from the concentrated leukocyte flowstream. Filtrate PRP collected from an optimally-designed CIF device typically showed a ~1000-fold (i.e. 99.9%) reduction in leukocyte concentration, while recovering >80% of the original platelets, at volumetric throughputs of ~1 mL/min. These results suggest that the CIF approach will enable users in many fields to now apply the advantages of microfluidic devices to particle separation, even for applications requiring macroscale flowrates.

  1. Single Cell Analysis of Leukocyte Protease Activity Using Integrated Continuous-Flow Microfluidics.

    Science.gov (United States)

    Jing, Tengyang; Lai, Zhangxing; Wu, Lidan; Han, Jongyoon; Lim, Chwee Teck; Chen, Chia-Hung

    2016-12-06

    Leukocytes are the essential cells of the immune system that protect the human body against bacteria, viruses, and other foreign invaders. Secretory products of individual leukocytes, such as matrix metalloproteinases (MMPs) and a disintegrin and metalloproteinase (ADAMs), are critical for regulating the inflammatory response and mediating host defense. Conventional single cell analytical methods, such as flow cytometry for cellular surface biomarker studies, are insufficient for performing functional assays of the protease activity of individual leukocytes. Here, an integrated continuous-flow microfluidic assay is developed to effectively detect secretory protease activity of individual viable leukocytes. Leukocytes in blood are first washed on-chip with defined buffer to remove background activity, followed by encapsulating individual leukocytes with protease sensors in water-in-oil droplets and incubating for 1 h to measure protease secretion. With this design, single leukocyte protease profiles under naive and phorbol 12-myristate 13-acetate (PMA)-stimulated conditions are reliably measured. It is found that PMA treatment not only elevates the average protease activity level but also reduces the cellular heterogeneity in protease secretion, which is important in understanding immune capability and the disease condition of individual patients.

  2. A multiscale SPH particle model of the near-wall dynamics of leukocytes in flow.

    Science.gov (United States)

    Gholami, Babak; Comerford, Andrew; Ellero, Marco

    2014-01-01

    A novel multiscale Lagrangian particle solver based on SPH is developed with the intended application of leukocyte transport in large arteries. In such arteries, the transport of leukocytes and red blood cells can be divided into two distinct regions: the bulk flow and the near-wall region. In the bulk flow, the transport can be modeled on a continuum basis as the transport of passive scalar concentrations. Whereas in the near-wall region, specific particle tracking of the leukocytes is required and lubrication forces need to be separately taken into account. Because of large separation of spatio-temporal scales involved in the problem, simulations of red blood cells and leukocytes are handled separately. In order to take the exchange of leukocytes between the bulk fluid and the near-wall region into account, solutions are communicated through coupling of conserved quantities at the interface between these regions. Because the particle tracking is limited to those leukocytes lying in the near-wall region only, our approach brings considerable speedup to the simulation of leukocyte circulation in a test geometry of a backward-facing step, which encompasses many flow features observed in vivo.

  3. Effect of leukocyte filtration on the P-selectin expression of apheresis platelets.

    Science.gov (United States)

    Xie, Z T; Chen, C; Zhang, S H; Yang, H M; Tao, Z H

    2015-06-01

    The aim of this study was to investigate the effect of leukocyte filtration on the P-selectin (CD62P) surface expression of apheresis platelets during the retention period. Ten bags of apheresis platelets stored for 1 day (0-24 h) and 10 bags of apheresis platelets stored for 2 days (24-48 h) were used for leukocyte filtration (experimental group). Ten bags of apheresis platelets with the corresponding retention periods but without filtration were used as a negative control (control group). Thereafter, 100 μL of platelet suspensions from apheresis platelets with or without leukocyte filtration were sampled before and after leukocyte filtration for the detection of CD62P surface expression by flow cytometry. No statistical difference in the CD62P surface expression of apheresis platelets was observed before and after leukocyte filtration (P > 0.05), neither did the CD62P surface expression exhibit any change among the different retention periods. Leukocyte filtration does not affect the CD62P surface expression of apheresis platelets stored for up to 2 days, which indicates that leukocyte filtration does not damage the activation of apheresis platelets within the retention period.

  4. A high-throughput microfluidic approach for 1000-fold leukocyte reduction of platelet-rich plasma.

    Science.gov (United States)

    Xia, Hui; Strachan, Briony C; Gifford, Sean C; Shevkoplyas, Sergey S

    2016-10-24

    Leukocyte reduction of donated blood products substantially reduces the risk of a number of transfusion-related complications. Current 'leukoreduction' filters operate by trapping leukocytes within specialized filtration material, while allowing desired blood components to pass through. However, the continuous release of inflammatory cytokines from the retained leukocytes, as well as the potential for platelet activation and clogging, are significant drawbacks of conventional 'dead end' filtration. To address these limitations, here we demonstrate our newly-developed 'controlled incremental filtration' (CIF) approach to perform high-throughput microfluidic removal of leukocytes from platelet-rich plasma (PRP) in a continuous flow regime. Leukocytes are separated from platelets within the PRP by progressively syphoning clarified PRP away from the concentrated leukocyte flowstream. Filtrate PRP collected from an optimally-designed CIF device typically showed a ~1000-fold (i.e. 99.9%) reduction in leukocyte concentration, while recovering >80% of the original platelets, at volumetric throughputs of ~1 mL/min. These results suggest that the CIF approach will enable users in many fields to now apply the advantages of microfluidic devices to particle separation, even for applications requiring macroscale flowrates.

  5. Distribution of leukocyte subtypes in the sheep ovary after laser drilling.

    Science.gov (United States)

    Tozawa, H; Brännström, M; Petrucco, O; Walker, S; Chambers, H; Pascoe, V; Norman, R J

    1995-03-01

    The distribution of leukocyte subtypes in the sheep ovary following laser drilling of the ovarian capsule was examined to understand a possible mechanism by which this treatment promotes ovulation in polycystic ovarian syndrome. Ovaries were removed from sheep at different time-points following laparoscopic laser drilling and immunohistochemical detection of leukocyte subtypes, using specific monoclonal antibodies; standard histological staining was performed. Migration of leukocytes into the laser-drilled site was observed as early as 6 h after laser drilling and the total number of leukocytes in the site was found to increase up to the 12th day after surgery. In the earlier period, polymorphonuclear leukocytes were the dominant leukocyte subtypes, while macrophages and lymphocytes were the major cellular components on the 12th day and later. These results show that the tissue changes in the ovary following laser drilling are consistent with a local inflammatory reaction. The prolonged appearance of numerous macrophages following the acute inflammatory phase could lead to the secretion of cytokines and other substances suggested to be important in promoting ovulation. These data indicate that part of the effectiveness of the laser drilling in polycystic ovarian syndrome may be attributable to the secretory products of these leukocytes.

  6. Downhill running and exercise in hot environments increase leukocyte Hsp72 (HSPA1A) and Hsp90α (HSPC1) gene transcripts.

    Science.gov (United States)

    Tuttle, James A; Castle, Paul C; Metcalfe, Alan J; Midgley, Adrian W; Taylor, Lee; Lewis, Mark P

    2015-04-15

    Stressors within humans and other species activate Hsp72 and Hsp90α mRNA transcription, although it is unclear which environmental temperature or treadmill gradient induces the largest increase. To determine the optimal stressor for priming the Hsp system, physically active but not heat-acclimated participants (19.8 ± 1.9 and 20.9 ± 3.6 yr) exercised at lactate threshold in either temperate (20°C, 50% relative humidity; RH) or hot (30°C, 50% RH) environmental conditions. Within each condition, participants completed a flat running (temperate flat or hot flat) and a downhill running (temperate downhill or hot downhill) experimental trial in a randomized counterbalanced order separated by at least 7 days. Venous blood samples were taken immediately before (basal), immediately after exercise, and 3 and 24 h postexercise. RNA was extracted from leukocytes and RT-quantitative PCR conducted to determine Hsp72 and Hsp90α mRNA relative expression. Leukocyte Hsp72 mRNA was increased immediately after exercise following downhill running (1.9 ± 0.9-fold) compared with flat running (1.3 ± 0.4-fold; P = 0.001) and in hot (1.9 ± 0.6-fold) compared with temperate conditions (1.1 ± 0.5-fold; P = 0.003). Leukocyte Hsp90α mRNA increased immediately after exercise following downhill running (1.4 ± 0.8-fold) compared with flat running (0.9 ± 0.6-fold; P = 0.002) and in hot (1.6 ± 1.0-fold) compared with temperate conditions (0.9 ± 0.6-fold; P = 0.003). Downhill running and exercise in hot conditions induced the largest stimuli for leukocyte Hsp72 and Hsp90α mRNA increases. Copyright © 2015 the American Physiological Society.

  7. Is Chronic Asthma Associated with Shorter Leukocyte Telomere Length at Midlife?

    Science.gov (United States)

    Shalev, Idan; Sears, Malcolm R.; Hancox, Robert J.; Lee Harrington, Hona; Houts, Renate; Moffitt, Terrie E.; Sugden, Karen; Williams, Benjamin; Poulton, Richie; Caspi, Avshalom

    2014-01-01

    Rationale: Asthma is prospectively associated with age-related chronic diseases and mortality, suggesting the hypothesis that asthma may relate to a general, multisystem phenotype of accelerated aging. Objectives: To test whether chronic asthma is associated with a proposed biomarker of accelerated aging, leukocyte telomere length. Methods: Asthma was ascertained prospectively in the Dunedin Multidisciplinary Health and Development Study cohort (n = 1,037) at nine in-person assessments spanning ages 9–38 years. Leukocyte telomere length was measured at ages 26 and 38 years. Asthma was classified as life-course-persistent, childhood-onset not meeting criteria for persistence, and adolescent/adult-onset. We tested associations between asthma and leukocyte telomere length using regression models. We tested for confounding of asthma-leukocyte telomere length associations using covariate adjustment. We tested serum C-reactive protein and white blood cell counts as potential mediators of asthma-leukocyte telomere length associations. Measurements and Main Results: Study members with life-course-persistent asthma had shorter leukocyte telomere length as compared with sex- and age-matched peers with no reported asthma. In contrast, leukocyte telomere length in study members with childhood-onset and adolescent/adult-onset asthma was not different from leukocyte telomere length in peers with no reported asthma. Adjustment for life histories of obesity and smoking did not change results. Study members with life-course-persistent asthma had elevated blood eosinophil counts. Blood eosinophil count mediated 29% of the life-course-persistent asthma-leukocyte telomere length association. Conclusions: Life-course-persistent asthma is related to a proposed biomarker of accelerated aging, possibly via systemic eosinophilic inflammation. Life histories of asthma can inform studies of aging. PMID:24956257

  8. Meisoindigo, but not its core chemical structure indirubin, inhibits zebrafish interstitial leukocyte chemotactic migration.

    Science.gov (United States)

    Ye, Baixin; Xiong, Xiaoxing; Deng, Xu; Gu, Lijuan; Wang, Qiongyu; Zeng, Zhi; Gao, Xiang; Gao, Qingping; Wang, Yueying

    2017-12-01

    Inflammatory disease is a big threat to human health. Leukocyte chemotactic migration is required for efficient inflammatory response. Inhibition of leukocyte chemotactic migration to the inflammatory site has been shown to provide therapeutic targets for treating inflammatory diseases. Our study was designed to discover effective and safe compounds that can inhibit leukocyte chemotactic migration, thus providing possible novel therapeutic strategy for treating inflammatory diseases. In this study, we used transgenic zebrafish model (Tg:zlyz-EGFP line) to visualize the process of leukocyte chemotactic migration. Then, we used this model to screen the hit compound and evaluate its biological activity on leukocyte chemotactic migration. Furthermore, western blot analysis was performed to evaluate the effect of the hit compound on the AKT or ERK-mediated pathway, which plays an important role in leukocyte chemotactic migration. In this study, using zebrafish-based chemical screening, we identified that the hit compound meisoindigo (25 μM, 50 μM, 75 μM) can significantly inhibit zebrafish leukocyte chemotactic migration in a dose-dependent manner (p = 0.01, p = 0.0006, p migration (p = 0.43). Furthermore, our results unexpectedly showed that indirubin, the core structure of meisoindigo, had no significant effect on zebrafish leukocyte chemotactic migration (p = 0.6001). Additionally, our results revealed that meisoindigo exerts no effect on the Akt or Erk-mediated signalling pathway. Our results suggest that meisoindigo, but not indirubin, is effective for inhibiting leukocyte chemotactic migration, thus providing a potential therapeutic agent for treating inflammatory diseases.

  9. Deformation mechanism of leukocyte adhering to vascular surface under steady shear flow

    Institute of Scientific and Technical Information of China (English)

    LIU; Xiaoheng; WANG; Xiong; YIN; Hongmei; CHEN; Huaiqing

    2004-01-01

    The adhesion of leukocytes to vascular surface is an important biomedical problem and has drawn extensive attention. In this study, we propose a compound drop model to simulate a leukocyte with a nucleus adhering to the surface of blood vessel under steady shear flow. A two-dimensional computational fluid dynamics (CFD) is conducted to determine the local distribution of pressure on the surface of the adherent model cell. By introducing the parameter of deformation index (DI), we investigate the deformation of the leukocyte and its nucleus under controlled conditions. Our numerical results show that: (i) the leukocyte is capable of deformation under external exposed flow field. The deformation index increases with initial contact angle and Reynolds number of external exposed flow. (ii) The nucleus deforms with the cell, and the deformation index of the leukocyte is greater than that of the nucleus. The leukocyte is more deformable while the nucleus is more capable of resisting external shear flow. (iii) The leukocyte and the nucleus are not able to deform infinitely with the increase of Reynolds number because the deformation index reaches a maximum. (iv) Pressure distribution confirms that there exists a region downstream of the cell, which produces high pressure to retard continuous deformation and provide a positive lift force on the cell. Meanwhile, we have measured the deformation of human leukocytes exposed to shear flow by using a flow chamber system. We found that the numerical results are well consistent with those of experiment. We conclude that the nucleus with high viscosity plays a particular role in leukocyte deformation.

  10. Longer leukocyte telomere length in Costa Rica's Nicoya Peninsula: a population-based study.

    Science.gov (United States)

    Rehkopf, David H; Dow, William H; Rosero-Bixby, Luis; Lin, Jue; Epel, Elissa S; Blackburn, Elizabeth H

    2013-11-01

    Studies in humans suggest that leukocyte telomere length may act as a marker of biological aging. We investigated whether individuals in the Nicoya region of Costa Rica, known for exceptional longevity, had longer telomere length than those in other parts of the country. After controlling for age, age squared, rurality, rainy season and gender, the mean leukocyte telomere length in Nicoya was substantially longer (81 base pairs, ptelomere length that characterizes this unique region does not appear to be explainable by traditional behavioral and biological risk factors. More detailed examination of mean leukocyte telomere length by age shows that the regional telomere length difference declines at older ages.

  11. Increased thymidylate synthase mRNA concentration in blood leukocytes following an experimental stressor

    DEFF Research Database (Denmark)

    Ehrnrooth, Eva; Zacharia, Robert; Svendsen, Gunner

    2002-01-01

    to a computerized mental stressor; (2) relaxation, and (3) control. Measurements included TS mRNA levels, total leukocyte number, leukocyte subtypes, and serum cortisol before (baseline), immediately after, and 1 h after each experimental condition. RESULTS: While no significant differences were found between...... in percentage of neutrophil cells after stress. CONCLUSION: The results suggest that TS mRNA levels in peripheral leukocytes may be sensitive to mental stress and confirm previous findings indicating that subjects scoring high on the personality trait of absorption exhibit greater physiological stress...

  12. Study of terahertz-radiation-induced DNA damage in human blood leukocytes

    Energy Technology Data Exchange (ETDEWEB)

    Angeluts, A A; Esaulkov, M N; Kosareva, O G; Solyankin, P M; Shkurinov, A P [International Laser Center, M. V. Lomonosov Moscow State University, Moscow (Russian Federation); Gapeyev, A B; Pashovkin, T N [Institute of Cell Biophysics, Russian Academy of Sciences, Pushchino, Moscow Region (Russian Federation); Matyunin, S N [Section of Applied Problems at the Presidium of the Russian Academy of Sciences, Moscow (Russian Federation); Nazarov, M M [Institute on Laser and Information Technologies, Russian Academy of Sciences, Shatura, Moscow Region (Russian Federation); Cherkasova, O P [Institute of Laser Physics, Siberian Branch, Russian Academy of Sciences, Novosibirsk (Russian Federation)

    2014-03-28

    We have carried out the studies aimed at assessing the effect of terahertz radiation on DNA molecules in human blood leukocytes. Genotoxic testing of terahertz radiation was performed in three different oscillation regimes, the blood leukocytes from healthy donors being irradiated for 20 minutes with the mean intensity of 8 – 200 μW cm{sup -2} within the frequency range of 0.1 – 6.5 THz. Using the comet assay it is shown that in the selected regimes such radiation does not induce a direct DNA damage in viable human blood leukocytes. (biophotonics)

  13. Leukocytes respiratory burst activity as indicator of innate immunity of pacu Piaractus mesopotamicus

    Directory of Open Access Journals (Sweden)

    JD Biller-Takahashi

    Full Text Available The present study evaluated the assay to quantify the respiratory burst activity of blood leukocytes of pacu as an indicator of the innate immune system, using the reduction of nitroblue tetrazolium (NBT to formazan as a measure of the production of reactive oxygen species (ROS. In order to assess the accuracy of the assay, fish were challenged by Aeromonas hydrophila and sampled one week after challenge. The A. hydrophila infection increased the leukocyte respiratory burst activity. The protocol showed a reliable and easy assay, appropriate to determine the respiratory burst activity of blood leukocytes of pacu, a neotropical fish, in the present experimental conditions.

  14. Efficiency of application original preservatives for conservation leukocytes at -40 °C

    Directory of Open Access Journals (Sweden)

    Utemov S.V.

    2011-12-01

    Full Text Available When frozen leukocytes exposed to harmful factors of the complex, due to their complex cellular structure and high metabolism. Cryopreservatives allow to avoid damages, but most of which are toxic. The aim of the present was to compare the efficacy of application of two non-toxic solutions for conservation of leukocytes at -40°С for 1 day. It was show that solution containing cryoprotector mixed action (a derivative of urea and antihypoxant (sodium fumarate is most effective in preserving the functional activity of leukocytes.

  15. [Anti-inflammatory action of a group of plant extracts].

    Science.gov (United States)

    Shipochliev, T; Dimitrov, A; Aleksandrova, E

    1981-01-01

    Use was made of Wistar albino rats in which an inflammation was induced via the simultaneous injection of caraginan and prostaglandin E1 in order to evaluate the antiinflammatory activity of 6 freeze dried plant extracts. It was found that with such model of inflammation the inflammatory effect of caraginan was strongly enhanced, which was accompanied by the rapid and prolific white blood cell extravasates. The freeze-dried extracts of St. John's-wort (Hypericum perforatum L.), potmarigold calendula (Calendula officinalis L.), camomile (Matricaria chamomilla L.) and plantain (Plantago lanceolata L. et Pl. major L.) were found to suppress both the inflammatory effect and the leukocyte infiltration. The extracts of symphytum (Symphytum officinale L.) and those of flax seed (Linum usitatissimum L.) did not inhibit the inflammation, however, they suppressed the leukocyte infiltration at the 3rd and 4th hour of the induced inflammation.

  16. 高通量透析器在维持性血液透析患者中的应用及护理%Clinical application of high flux dialyzer in hemodialysis

    Institute of Scientific and Technical Information of China (English)

    王文静

    2015-01-01

    To evaluate the effect of high flux dialyzer on hemodialysis.Methods: 36 cases of hemodialysis took high flux hemodialysis treatment,and were given the targeted nursing measures to monitor first and 3 months of the improvement.Results: after using,greatly improved,the quality of life of patients improved significantly.Conclusion: high flux hemodialysis and the corresponding nursing measures can significantly improve the quality of life with maintenance hemodialysis.%探讨维持性血液透析患者使用高通量透析器的护理方法及效果.方法:对我院36例透析患者进行高通量血液透析治疗,并给予针对性的护理措施,监测使用前和使用后3个月在饮食、精神、活动能力等方面的改善情况.结果:使用后比使用前在饮食、精神、活动能力等方面,有极大改善,患者生活质量明显改善.结论:高通量透析和相应的护理措施能明显改善维持性透析患者的生活质量.

  17. Removal of methadone by extended dialysis using a high cut-off dialyzer: implications for the treatment of overdose and for pain management in patients undergoing light chain removal.

    Science.gov (United States)

    Arelin, Viktor; Schmidt, Julius J; Kayser, Nathalie; Kühn-Velten, W Nikolaus; Suhling, Hendrik; Eden, Gabriele; Kielstein, Jan T

    2016-06-01

    The synthetic opioid methadone hydrochloride has a low molecular weight of 346 D, a high volume of distribution (4 - 7 L/kg), and is lipophilic. It is used as an analgesic and for the maintenance treatment of opiate dependence. In drug addicts, methadone is frequently involved in mixed intoxications that can lead to death. Here we present the case of a drug addict in whom a high cut-off dialysis membrane together with extended dialysis was used in the setting of suspected overdose and acute kidney injury. Although the observed dialyzer plasma clearance (31.5 mL/min) and reduction ratio (38%) were higher than previously reported for standard hemodialysis, the total amount of methadone in the spent dialysate after 1 extended dialysis session was quite low. Hence, even extended dialysis with a high cut-off membrane does not seem to offer a clinically relevant benefit in the setting of overdose for enhanced methadone removal. On the other hand, in patients undergoing high cut-off dialysis for the removal of light chains, methadone could still be used as an analgesic without an additional dose after high cut-off hemodialysis.

  18. Integrin activation by P-Rex1 is required for selectin-mediated slow leukocyte rolling and intravascular crawling.

    Science.gov (United States)

    Herter, Jan M; Rossaint, Jan; Block, Helena; Welch, Heidi; Zarbock, Alexander

    2013-03-21

    Integrin activation is essential for the function of leukocytes. Impaired integrin activation on leukocytes is the hallmark of the leukocyte adhesion deficiency syndrome in humans, characterized by impaired leukocyte recruitment and recurrent infections. In inflammation, leukocytes collect different signals during the contact with the microvasculature, which activate signaling pathways leading to integrin activation and leukocyte recruitment. We report the role of P-Rex1, a Rac-specific guanine nucleotide exchanging factor, in integrin activation and leukocyte recruitment. We find that P-Rex1 is required for inducing selectin-mediated lymphocyte function-associated antigen-1 (LFA-1) extension that corresponds to intermediate affinity and induces slow leukocyte rolling, whereas P-Rex1 is not involved in the induction of the high-affinity conformation of LFA-1 obligatory for leukocyte arrest. Furthermore, we demonstrate that P-Rex1 is involved in Mac-1-dependent intravascular crawling. In vivo, both LFA-1-dependent slow rolling and Mac-1-dependent crawling are defective in P-Rex1(-/-) leukocytes, whereas chemokine-induced arrest and postadhesion strengthening remain intact in P-Rex1-deficient leukocytes. Rac1 is involved in E-selectin-mediated slow rolling and crawling. In vivo, in an ischemia-reperfusion-induced model of acute kidney injury, abolished selectin-mediated integrin activation contributed to decreased neutrophil recruitment and reduced kidney damage in P-Rex1-deficient mice. We conclude that P-Rex1 serves distinct functions in LFA-1 and Mac-1 activation.

  19. Characterization of Leukocyte Formin FMNL1 Expression in Human Tissues

    Science.gov (United States)

    Heuser, Vanina D.; Iljin, Kristiina; Kampf, Caroline; Uhlen, Mathias; Carpén, Olli

    2014-01-01

    Formins are cytoskeleton regulating proteins characterized by a common FH2 structural domain. As key players in the assembly of actin filaments, formins direct dynamic cytoskeletal processes that influence cell shape, movement and adhesion. The large number of formin genes, fifteen in the human, suggests distinct tasks and expression patterns for individual family members, in addition to overlapping functions. Several formins have been associated with invasive cell properties in experimental models, linking them to cancer biology. One example is FMNL1, which is considered to be a leukocyte formin and is known to be overexpressed in lymphomas. Studies on FMNL1 and many other formins have been hampered by a lack of research tools, especially antibodies suitable for staining paraffin-embedded formalin-fixed tissues. Here we characterize, using bioinformatics tools and a validated antibody, the expression pattern of FMNL1 in human tissues and study its subcellular distribution. Our results indicate that FMNL1 expression is not restricted to hematopoietic tissues and that neoexpression of FMNL1 can be seen in epithelial cancer. PMID:24700756

  20. Acoustic and photoacoustic microscopy imaging of single leukocytes

    Science.gov (United States)

    Strohm, Eric M.; Moore, Michael J.; Kolios, Michael C.

    2016-03-01

    An acoustic/photoacoustic microscope was used to create micrometer resolution images of stained cells from a blood smear. Pulse echo ultrasound images were made using a 1000 MHz transducer with 1 μm resolution. Photoacoustic images were made using a fiber coupled 532 nm laser, where energy losses through stimulated Raman scattering enabled output wavelengths from 532 nm to 620 nm. The laser was focused onto the sample using a 20x objective, and the laser spot co-aligned with the 1000 MHz transducer opposite the laser. The blood smear was stained with Wright-Giemsa, a common metachromatic dye that differentially stains the cellular components for visual identification. A neutrophil, lymphocyte and a monocyte were imaged using acoustic and photoacoustic microscopy at two different wavelengths, 532 nm and 600 nm. Unique features in each imaging modality enabled identification of the different cell types. This imaging method provides a new way of imaging stained leukocytes, with applications towards identifying and differentiating cell types, and detecting disease at the single cell level.

  1. Current opinion on human leukocyte antigen-G in China

    Institute of Scientific and Technical Information of China (English)

    YAN Wei-hua; LIN Ai-fen

    2007-01-01

    @@ Since discovery and cloning of the non-classical human leukocyte antigen (HLA) class Ⅰ antigen HLA-G by Geraghty et al1 in 1987, a large number of studies have been carried out. HLA-G has a low polymorphism, limited distribution to normal tissues and seven isoforms resulting from its primary mRNA alternative splicing.2 HLA-G expression was first found on the extravillous cytotrophoblasts, at the fetal-maternal interface during normal pregnancy, which lacks the expression of HLA-A, -B and HLA Ⅱ antigens. Initial studies on HLA-G mainly addressed its function in fetal-maternal immunotolerance.3 Two decades later,HLA-G is now considered to be a very important immune molecule which plays a vital immune inhibitory role in the context of reproduction, oncology, transplantation,infection and also in autoimmune disease.4 A number of Chinese research teams are interested in, and have contributed to, the publication of more than 80 peer-reviewed articles and reviews on HLA-G over the past ten years. We summarize the key points in this field that were presented and discussed by them.

  2. Role of bacteria in leukocyte adhesion deficiency-associated periodontitis.

    Science.gov (United States)

    Hajishengallis, George; Moutsopoulos, Niki M

    2016-05-01

    Leukocyte adhesion deficiency Type I (LAD-I)-associated periodontitis is an aggressive form of inflammatory bone loss that has been historically attributed to lack of neutrophil surveillance of the periodontal infection. However, this form of periodontitis has proven unresponsive to antibiotics and/or mechanical removal of the tooth-associated biofilm. Recent studies in LAD-I patients and relevant animal models have shown that the fundamental cause of LAD-I periodontitis involves dysregulation of a granulopoietic cytokine cascade. This cascade includes interleukin IL-23 (IL-23) and IL-17 that drive inflammatory bone loss in LAD-I patients and animal models and, moreover, foster a nutritionally favorable environment for bacterial growth and development of a compositionally unique microbiome. Although the lack of neutrophil surveillance in the periodontal pockets might be expected to lead to uncontrolled bacterial invasion of the underlying connective tissue, microbiological analyses of gingival biopsies from LAD-I patients did not reveal tissue-invasive infection. However, bacterial lipopolysaccharide was shown to translocate into the lesions of LAD-I periodontitis. It is concluded that the bacteria serve as initial triggers for local immunopathology through translocation of bacterial products into the underlying tissues where they unleash the dysregulated IL-23-IL-17 axis. Subsequently, the IL-23/IL-17 inflammatory response sustains and shapes a unique local microbiome which, in turn, can further exacerbate inflammation and bone loss in the susceptible host. Copyright © 2015 Elsevier Ltd. All rights reserved.

  3. Swine leukocyte antigen (SLA) diversity in Sinclair and Hanford swine.

    Science.gov (United States)

    Ho, Chak-Sum; Martens, Gregory W; Amoss, Max S; Gomez-Raya, Luis; Beattie, Craig W; Smith, Douglas M

    2010-03-01

    The swine leukocyte antigen (SLA) haplotype B is associated with increased penetrance of the tumor traits in Sinclair swine cutaneous melanoma (SSCM). We established a series of SinclairxHanford swine crosses to facilitate genetic mapping of the tumor-associated loci. In this study, the SLA diversity in the founding animals was characterized for effective selection of maximum tumor penetrance in the pedigrees. Using the sequence-based typing (SBT) method we identified a total of 29 alleles at five polymorphic SLA loci (SLA-1, SLA-3, SLA-2, DRB1 and DQB1) representing six class I and five class II haplotypes. We subsequently developed a rapid PCR-based typing assay using sequence-specific primers (PCR-SSP) to efficiently follow the SLA types of the crossbred progeny. In a total of 469 animals we identified three crossovers within the class I region and three between the class I and class II regions, which corresponded to recombination frequencies of 0.39% and 0.56%, respectively. We also confirmed the presence of two expressed SLA-1 loci in three of the class I haplotypes and were able to determine the relative chromosomal arrangement of the duplicated loci in two haplotypes. This study furthers our understanding of the allelic architecture and polymorphism of the SLA system and will facilitate the mapping of loci associated with the expression of SSCM. Copyright 2009 Elsevier Ltd. All rights reserved.

  4. Role of salivary leukocyte protease inhibitor in periodontal disease progression

    Directory of Open Access Journals (Sweden)

    Pateel Deepak

    2010-01-01

    Full Text Available Context: Proteases play a major role in the tissue destruction involved in periodontal disease. It is known that the balance between proteases and their inhibitors is a major determinant in maintaining tissue integrity. The association between the proteases and periodontitis is well established, but not many studies have been carried out to know the role played by a protease inhibitor like salivary leukocyte protease inhibitor (SLPI in periodontitis. Aim: The aim of the present study was to correlate SLPI with periodontitis. Settings and Design: Case-control study. Materials and Methods: Seventy-five clinically confirmed cases of periodontitis and 20 controls were included in the study. A detailed case history and periodontal index (PI were recorded. Two milliliters of unstimulated saliva samples was obtained and subjected to quantification of SLPI leaves using SLPI in enzyme-linked immunosorbent assay (ELISA kit. Based on the periodontal index score of the individuals, the cases and controls were divided into groups A, B and C, and the obtained SLPI levels were compared among the groups. Statistical Analysis: Mann-Whitney U test and correlation coefficient test. Results: The results showed that in the initial stages of periodontitis there is a tendency of SLPI levels to be raised. The SLPI levels were found to be reduced in the terminal stages of periodontitis. Conclusion: It appears that SLPI accumulates in the local environment, at least in the initial stages of the periodontal disease, probably to inhibit the action of increased elastic activity.

  5. HLA-Modeler: Automated Homology Modeling of Human Leukocyte Antigens

    Directory of Open Access Journals (Sweden)

    Shinji Amari

    2013-01-01

    Full Text Available The three-dimensional (3D structures of human leukocyte antigen (HLA molecules are indispensable for the studies on the functions at molecular level. We have developed a homology modeling system named HLA-modeler specialized in the HLA molecules. Segment matching algorithm is employed for modeling and the optimization of the model is carried out by use of the PFROSST force field considering the implicit solvent model. In order to efficiently construct the homology models, HLA-modeler uses a local database of the 3D structures of HLA molecules. The structure of the antigenic peptide-binding site is important for the function and the 3D structure is highly conserved between various alleles. HLA-modeler optimizes the use of this structural motif. The leave-one-out cross-validation using the crystal structures of class I and class II HLA molecules has demonstrated that the rmsds of nonhydrogen atoms of the sites between homology models and crystal structures are less than 1.0 Å in most cases. The results have indicated that the 3D structures of the antigenic peptide-binding sites can be reproduced by HLA-modeler at the level almost corresponding to the crystal structures.

  6. HLA-Modeler: Automated Homology Modeling of Human Leukocyte Antigens.

    Science.gov (United States)

    Amari, Shinji; Kataoka, Ryoichi; Ikegami, Takashi; Hirayama, Noriaki

    2013-01-01

    The three-dimensional (3D) structures of human leukocyte antigen (HLA) molecules are indispensable for the studies on the functions at molecular level. We have developed a homology modeling system named HLA-modeler specialized in the HLA molecules. Segment matching algorithm is employed for modeling and the optimization of the model is carried out by use of the PFROSST force field considering the implicit solvent model. In order to efficiently construct the homology models, HLA-modeler uses a local database of the 3D structures of HLA molecules. The structure of the antigenic peptide-binding site is important for the function and the 3D structure is highly conserved between various alleles. HLA-modeler optimizes the use of this structural motif. The leave-one-out cross-validation using the crystal structures of class I and class II HLA molecules has demonstrated that the rmsds of nonhydrogen atoms of the sites between homology models and crystal structures are less than 1.0 Å in most cases. The results have indicated that the 3D structures of the antigenic peptide-binding sites can be reproduced by HLA-modeler at the level almost corresponding to the crystal structures.

  7. Identifying coevolutionary patterns in human leukocyte antigen (HLA) molecules.

    Science.gov (United States)

    Jiang, Xiaowei; Fares, Mario A

    2010-05-01

    The antigenic peptide, major histocompatibility complex molecule (MHC; also called human leukocyte antigen, HLA), coreceptor CD8, or CD4 and T-cell receptor (TCR) function as a complex to initiate effectors' mechanisms of the immune system. The tight functional and physical interaction among these molecules may have involved strong coevolution links among domains within and between proteins. Despite the importance of unraveling such dependencies to understand the arms race of host-pathogen interaction, no previous studies have aimed at achieving such an objective. Here, we perform an exhaustive coevolution analysis and show that indeed such dependencies are strongly shaping the evolution and probably the function of these molecules. We identify intramolecular coevolution in HLA class I and II at domains important for their immune activity. Most of the amino acid sites identified to be coevolving in HLAI have been also detected to undergo positive Darwinian selection highlighting therefore their adaptive value. We also identify coevolution among antigen-binding pockets (P1-P9) and among these and TCR-binding sites. Conversely to HLAI, coevolution is weaker in HLAII. Our results support that such coevolutionary patterns are due to selective pressures of host-pathogen coevolution and cooperative binding of TCRs, antigenic peptides, and CD8/CD4 to HLAI and HLAII.

  8. Automatic leukocyte nucleus segmentation by intuitionistic fuzzy divergence based thresholding.

    Science.gov (United States)

    Jati, Arindam; Singh, Garima; Mukherjee, Rashmi; Ghosh, Madhumala; Konar, Amit; Chakraborty, Chandan; Nagar, Atulya K

    2014-03-01

    The paper proposes a robust approach to automatic segmentation of leukocyte's nucleus from microscopic blood smear images under normal as well as noisy environment by employing a new exponential intuitionistic fuzzy divergence based thresholding technique. The algorithm minimizes the divergence between the actual image and the ideally thresholded image to search for the final threshold. A new divergence formula based on exponential intuitionistic fuzzy entropy has been proposed. Further, to increase its noise handling capacity, a neighborhood-based membership function for the image pixels has been designed. The proposed scheme has been applied on 110 normal and 54 leukemia (chronic myelogenous leukemia) affected blood samples. The nucleus segmentation results have been validated by three expert hematologists. The algorithm achieves an average segmentation accuracy of 98.52% in noise-free environment. It beats the competitor algorithms in terms of several other metrics. The proposed scheme with neighborhood based membership function outperforms the competitor algorithms in terms of segmentation accuracy under noisy environment. It achieves 93.90% and 94.93% accuracies for Speckle and Gaussian noises, respectively. The average area under the ROC curves comes out to be 0.9514 in noisy conditions, which proves the robustness of the proposed algorithm.

  9. Leukocyte telomere length: Effects of schizophrenia, age, and gender.

    Science.gov (United States)

    Wolkowitz, Owen M; Jeste, Dilip V; Martin, Averria Sirkin; Lin, Jue; Daly, Rebecca E; Reuter, Chase; Kraemer, Helena

    2017-02-01

    Schizophrenia is linked with early medical comorbidity and mortality. These observations indicate possible "accelerated biological aging" in schizophrenia, although prior findings are mixed, and few such studies have examined the role of gender. One putative marker of biological aging is leukocyte telomere length (LTL), which typically shortens with age. We assessed LTL in phenotypically well characterized 134 individuals with schizophrenia (60 women and 74 men) and 123 healthy comparison subjects (HCs) (66 women and 57 men), aged 26 to 65 years. Overall, LTL was inversely associated with age (t(249) = -6.2, p schizophrenia and HC groups (t(249) = 2.48, p = 0.014), with women having longer LTL than men, and a significant gender X diagnosis effect (t(249) = 2.43, p = 0.016) - at the average sample age, women with schizophrenia had shorter LTL than HC women. Gender, not the diagnosis of schizophrenia, was the major factor involved with LTL shortening across the age range studied. We discuss the constraints of a cross-sectional design and other methodological issues, and indicate future directions. Understanding the impact of schizophrenia on biological aging will require separate evaluations in men and women. Published by Elsevier Ltd.

  10. Relationship between vitamin D status and leukocytes in hospitalised cats.

    Science.gov (United States)

    Titmarsh, Helen F; Cartwright, Jennifer A; Kilpatrick, Scott; Gaylor, Donna; Milne, Elspeth M; Berry, Jacqueline L; Bommer, Nicholas X; Gunn-Moore, Danièlle; Reed, Nicola; Handel, Ian; Mellanby, Richard J

    2017-04-01

    Objectives Vitamin D deficiency, as assessed by serum 25-hydroxyvitamin D (25[OH]D) concentrations, has been linked to markers of systemic inflammation in human and canine medicine. However, the relationship between vitamin D status and inflammation has not been previously investigated in cats. The aim of this study was to examine the relationship between serum 25(OH)D concentrations and leukocyte counts in hospitalised sick cats. Methods Serum 25(OH)D concentrations and haematology profiles were measured in 170 consecutive hospitalised sick cats. A binary logistical regression model examined the relationship between serum 25(OH)D concentration, age, sex, breed and neutrophil, monocyte, eosinophil and lymphocyte counts. Results Cats with neutrophilia had lower serum 25(OH)D concentrations than cats with neutrophil concentrations below the upper limit of the reference interval (RI). There were no differences in serum 25(OH)D concentrations in cats with monocyte, lymphocyte or eosinophil counts above their respective RI compared with cats with counts below the upper limit of the RI. Conclusions and relevance Hospitalised cats with a neutrophil count above the RI had lower vitamin D status. There is a need to establish whether lower vitamin D status is a cause or consequence of increased neutrophil counts.

  11. Impact of Dehydroepiandrosterone Sulfate on Newborn Leukocyte Telomere Length

    Science.gov (United States)

    Liu, Han; Zhou, Guangdi; Chen, Qian; Ouyang, Fengxiu; Little, Julian; Zhang, Jun; Chen, Dan

    2017-01-01

    The newborn setting of leukocyte telomere length (LTL) likely has important implications for telomere dynamics over the lifespan. However, its determinants are poorly understood. Hormones play an important role during pregnancy and delivery. We hypothesized that exposure to hormones may impact the fetal telomere biology system. To test this hypothesis, cortisol, estradiol, dehydroepiandrosterone sulfate (DHEAS) and reactive oxygen species (ROS) were measured in cord blood of 821 newborns from a prospective study. After accounting for the effects of potential determinants of newborn LTL, a 10-fold increase in DHEAS concentration was associated with a 0.021 increase in T/S ratio of newborn LTL (95% confidence interval: 0.009–0.034, P = 0.0008). For newborns who fell in the lowest quartile of DHEAS level, the mean newborn LTL was estimated to be approximately 2.0% shorter than the newborns in the highest DHEAS concentration quartile (P = 0.0014). However, no association was found between newborn LTL and cortisol or estradiol. As expected, newborns with higher ROS level (ROS > 260 mol/L) had lower LTL compared to that with lower ROS level (ROS ≤ 260 mol/L) (P = 0.007). There was also an inverse relationship between DHEAS and ROS (P programming” effect on the newborn telomere biology system. PMID:28186106

  12. Leukocyte deformability: finite element modeling of large viscoelastic deformation.

    Science.gov (United States)

    Dong, C; Skalak, R

    1992-09-21

    An axisymmetric deformation of a viscoelastic sphere bounded by a prestressed elastic thin shell in response to external pressure is studied by a finite element method. The research is motivated by the need for understanding the passive behavior of human leukocytes (white blood cells) and interpreting extensive experimental data in terms of the mechanical properties. The cell at rest is modeled as a sphere consisting of a cortical prestressed shell with incompressible Maxwell fluid interior. A large-strain deformation theory is developed based on the proposed model. General non-linear, large strain constitutive relations for the cortical shell are derived by neglecting the bending stiffness. A representation of the constitutive equations in the form of an integral of strain history for the incompressible Maxwell interior is used in the formulation of numerical scheme. A finite element program is developed, in which a sliding boundary condition is imposed on all contact surfaces. The mathematical model developed is applied to evaluate experimental data of pipette tests and observations of blood flow.

  13. EDU pretreatment decreases polymorphonuclear leukocyte migration into rat lung airways.

    Science.gov (United States)

    Bassett, D J; Elbon, C L; Ishii, Y; Yang, H; Otterbein, L; Boswell, G A; Kerr, J S

    1994-07-01

    Pretreatment with the heterocyclic compound EDU (N-[2-(2-oxo-1-imidazolindinyl)ethyl]-N'-phenylurea) has previously been shown to reduce polymorphonuclear leukocyte (PMN) infiltration into the airways of ozone-exposed rats. The present study further examined the effects of 1 and 2 days EDU pretreatment on rat lung inflammatory responses by determining PMN infiltration in response to intratracheal instillation with the chemoattractant formyl-norleucine-leucine-phenylalanine (fNLP). Maximal recovery of PMNs by bronchoalveolar lavage was observed 4 hr after fNLP instillation with no alteration in the numbers of recoverable macrophages and lymphocytes. Although 1-day pretreatment with EDU did not affect PMN recovery from fNLP-instilled rat lungs, 2 days of EDU pretreatment prevented PMN infiltration as indicated by PMN recoveries that were similar to those obtained from saline-instilled lungs. Measurements of lung-marginated and interstitial pools of inflammatory cells using collagenase tissue digestion demonstrated no effect of 2 days EDU pretreatment. Although 2 days EDU pretreatment alone did not alter blood PMN content, lung permeability, and the lavage recoveries of inflammatory cells, blood PMN responses to chemotactic stimuli in vitro were impaired. In addition, EDU was shown to directly inhibit PMN chemotaxis and superoxide anion generation in vitro. These data demonstrated that EDU acts by interfering with PMN activation and migration rather than by decreasing PMN availability. EDU, by modulating the inflammatory response, represents a useful compound for preventing PMN-associated amplification of acute lung injuries.

  14. Leukocyte and leukocyte function

    Institute of Scientific and Technical Information of China (English)

    2008-01-01

    2008333 The clinical and laboratory features of acute promyelocytic leukemia: an analysis of 513 cases. LIANG Jianying(梁建英), et al. Dept Hematol, 1st Affili Hosp, Soochow Univ, Suzhou 215006. Chin J Interm Med 2008;47(5):389-392.Objective To investigate the clinical and laboratory features of acute promyelocytic leukemia(APL).Methods 513 APL patients in the last two decades were retrospectively analyzed in this research.We investigated the clinical features including age,sex,abnormality of

  15. Leukocyte and leukocyte function

    Institute of Scientific and Technical Information of China (English)

    1995-01-01

    950368 A study on the relationship hetween morphol-ogy and gene hepterogeneity in acute promyelocyticleukemia.XIONG Shumin(熊树民),et al.shanghaiHematol Instit & Ruijin Hosp,Shanghai,200025.ChinJ Intern Med 1995;34(3):165-168.Aucte promyelocytic leukemia (APL) can be treatedby all-trans retinoic acid (ATRA) with high completeremission rate.50 cases of APL diagnosed morphologi-cally were studied on their cytogenetics,molecular bi-ology and response to treatment with ATRA.Fortyfive cases showed chromosomal translocation t(5;17)and PML/RAR α fusion gene (PML+RARα+APL)

  16. Leukocyte and leukocyte function

    Institute of Scientific and Technical Information of China (English)

    1993-01-01

    930574 Phase II clinical trial of domestic ant-saerine in patients with acute leukemias.Cooper-ative Group headed by Blood dis Hosp,InstitHematol,CAMS,Tianjin,300020.Chin J In-tern Med 1993;32(2):80—83.One hundred and eighteen patients with acuteleukemias,including initial,relapsed and refrac-tory cases,were treated with domestic Am-sacrine(m—AMSA),single or in combinationwith other drugs.The total CR rate was 39.5%in ALL and 38.8% in ANLL and the responserate was 47.5% for both types of acuteleukemias.The CR rates of relapsed and refrac-

  17. Leukocyte and leukocyte function

    Institute of Scientific and Technical Information of China (English)

    2009-01-01

    2009228 A new prognostic stratification for patients with acute myeloid leukemia.JIANG Bo(姜波),et al.Instit Hematol & Blood Dis Hosp,CAMS & PUMC,Tianjin 300020.Chin J Intern Med,2009;48(4):316-320.

  18. Leukocyte and leukocyte function

    Institute of Scientific and Technical Information of China (English)

    2010-01-01

    2010367 The clinical characteristics of newly diagnosed acute myeloid leukemia patients with NPM1 mutation. ZHU Honghu(主鸿鹄),et al.Instit Hematol,People’ s Hosp,Peking Univ,Beijing 100044.Chin J Hematol 2010;31(5):315-318. Objective To investigate the clinical characteristics of newly diagnosed acute myeloid leukemia (AML) with NPM1 mutation.Methods

  19. Transfusion of Leukocyte-Depleted RBCs Is Independently Associated With Increased Morbidity After Pediatric Cardiac Surgery

    NARCIS (Netherlands)

    Kneyber, Martin C. J.; Grotenhuis, Femke; Berger, Rolf F. M.; Ebels, Tjark W.; Burgerhof, Johannes G. M.; Albers, Marcel J. I. J.

    Objective: To test the hypothesis that transfusion of leukocyte-depleted RBC preparations within the first 48 hours of PICU stay was independently associated with prolonged duration of mechanical ventilation, irrespective of surgery type and disease severity. Design: Retrospective, observational

  20. Leukocyte scintigraphy compared to intraoperative small bowel enteroscopy and laparotomy findings in Crohn's disease

    DEFF Research Database (Denmark)

    Almen, Sven; Granerus, Göran; Ström, Magnus

    2007-01-01

    Background: Leukocyte scintigraphy is a noninvasive investigation to assess inflammation. We evaluated the utility of labeled leukocytes to detect small bowel inflammation and disease complications in Crohn's disease and compared it to whole small bowel enteroscopy and laparotomy findings. Methods......: Scintigraphy with technetium-99m exametazime-labeled leukocytes was prospectively performed in 48 patients with Crohn's disease a few days before laparotomy; 41 also had an intraoperative small bowel enteroscopy. The same procedures were performed in 8 control patients. Independent grading of scans...... was compared with the results of enteroscopy and with surgical, histopathologic, and clinical data. Results: In the 8 control patients leukocyte scan, endoscopy, and histopathology were all negative for the small bowel. In patients with Crohn's disease and small bowel inflammation seen at enteroscopy and...

  1. HAEMATO-BIOCHEMICAL PROFILE AND MILK LEUKOCYTE COUNT IN SUBCLINICAL AND CLINICAL MASTITIS AFFECTED CROSSBRED CATTLE

    National Research Council Canada - National Science Library

    Sarvesha K; Satyanarayana M L; Narayanaswamy H D; Rao S; Yathiraj S; Isloor S; Mukartal S Y; Singh S V; Anuradha M E

    2017-01-01

    .... The SCM were diagnosed by California Mastitis Test (CMT) and electrical conductivity (EC) of milk. Blood and milk of mastitis infected crossbred cattle were analyzed for haemato-biochemical parameters and milk leukocyte count...

  2. In vitro phagocytosis of several Candida berkhout species by murine leukocytes.

    Science.gov (United States)

    Fontenla de Petrino, S E; Bibas Bonet de Jorrat, M E; Sirena, A

    1985-03-01

    In vitro phagocytosis of thirteen Candida berkhout species by rat leukocytes was studied to assess a possible correlation between pathogenicity and phagocytosis Yeast-leukocyte suspensions were mixed up for 3 h and phagocytic index, germ-tube formation and leukocyte candidacidal activity were evaluated. Highest values for phagocytosis were reached in all cases at the end of the first hour. Leukocyte candidacidal activity was absent. Only C. albicans produced germ-tubes. The various phagocytosis indices were determined depending on the Candida species assayed. Under these conditions, the more pathogenic species presented the lower indices of phagocytosis. It is determined that the in vitro phagocytic index may bear a close relationship with the pathogenicity of the Candida berkhout.

  3. Activated leukocyte cell adhesion molecule and prognosis in acute ischemic stroke

    DEFF Research Database (Denmark)

    Smedbakken, Linda; Jensen, Jesper K; Hallén, Jonas

    2011-01-01

    Biomarkers predicting mortality and functional outcome in stroke may be clinically helpful in identification of patients likely to benefit from intervention. Activated leukocyte cell adhesion molecule (ALCAM) is upregulated during neuroinflammation; we investigated whether ALCAM concentrations...

  4. Evaluation of leukocyte esterase and nitrite strip tests to detect spontaneous bacterial peritonitis in cirrhotic patients

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    AIM: To investigate the diagnostic efficacy of leukocyte esterase and nitrite reagent strips for bedside diagnosis of spontaneous bacterial peritonitis (SBP).METHODS: A total of 63 consecutive patients with cirrhotic ascites (38 male, 25 female) tested between April 2005 and July 2006 were included in the study. Bedside reagent strip testing was performed on ascitic fluid and the results compared to manual cell counting and ascitic fluid culture. SBP was defined as having a olymorphonuclear ascites count of ≥ 250/mm3.RESULTS: Fifteen samples showed SBP. The sensitivity,specificity, positive and negative predictive values of the leukocyte esterase reagent strips were; 93%, 100%, 100%, and 98%, respectively. The sensitivity,specificity, positive and negative predictive value of the nitrite reagent strips were 13%, 93%, 40%, and 77%, respectively. The combination of leukocyte esterase and nitrite reagents strips did not yield statistically significant effects on diagnostic accuracy. CONCLUSION: Leukocyte esterase reagent strips may provide a rapid, bedside diagnostic test for SBP.

  5. Syntenin-1 and ezrin proteins link activated leukocyte cell adhesion molecule to the actin cytoskeleton

    NARCIS (Netherlands)

    Tudor, C.; Riet, J. te; Eich, C.; Harkes, R.; Smisdom, N.; Bouhuijzen-Wenger, J.; Ameloot, M.; Holt, M.; Kanger, J.S.; Figdor, C.G.; Cambi, A.; Subramaniam, V.

    2014-01-01

    Activated leukocyte cell adhesion molecule (ALCAM) is a type I transmembrane protein member of the immunoglobulin superfamily of cell adhesion molecules. Involved in important pathophysiological processes such as the immune response, cancer metastasis, and neuronal development, ALCAM undergoes both

  6. Inflammation after Ischemic Stroke: The Role of Leukocytes and Glial Cells

    Science.gov (United States)

    Kim, Jong Youl; Park, Joohyun; Chang, Ji Young; Kim, Sa-Hyun

    2016-01-01

    The immune response after stroke is known to play a major role in ischemic brain pathobiology. The inflammatory signals released by immune mediators activated by brain injury sets off a complex series of biochemical and molecular events which have been increasingly recognized as a key contributor to neuronal cell death. The primary immune mediators involved are glial cells and infiltrating leukocytes, including neutrophils, monocytes and lymphocyte. After ischemic stroke, activation of glial cells and subsequent release of pro- and anti-inflammatory signals are important for modulating both neuronal cell damage and wound healing. Infiltrated leukocytes release inflammatory mediators into the site of the lesion, thereby exacerbating brain injury. This review describes how the roles of glial cells and circulating leukocytes are a double-edged sword for neuroinflammation by focusing on their detrimental and protective effects in ischemic stroke. Here, we will focus on underlying characterize of glial cells and leukocytes under inflammation after ischemic stroke.

  7. Microfluidic isolation of leukocytes from whole blood for phenotype and gene expression analysis.

    Science.gov (United States)

    Sethu, Palaniappan; Moldawer, Lyle L; Mindrinos, Michael N; Scumpia, Philip O; Tannahill, Cynthia L; Wilhelmy, Julie; Efron, Philip A; Brownstein, Bernard H; Tompkins, Ronald G; Toner, Mehmet

    2006-08-01

    Technologies that enable the isolation of cell subtypes from small samples of complex populations will greatly facilitate the implementation of proteomics and genomics to human diseases. Transcriptome analysis of blood requires the depletion of contaminating erythrocytes. We report an automated microfluidic device to rapidly deplete erythrocytes from whole blood via deionized water lysis and to collect enriched leukocytes for phenotype and genomic analyses. Starting with blood from healthy subjects, we demonstrate the utility of this microfluidic cassette and lysis protocol to prepare unstimulated leukocytes, and leukocytes stimulated ex vivo with Staphylococcal enterotoxin B, which mimics some of the cellular effects seen in patients with severe bacterial infections. Microarrays are used to assess the global gene expression response to enterotoxin B. The results demonstrate that this system can isolate unactivated leukocytes from small blood samples without any significant loss, which permits more information to be obtained from subsequent analysis, and will be readily applicable to clinical settings.

  8. Appearance of acute gouty arthritis on indium-111-labeled leukocyte scintigraphy

    Energy Technology Data Exchange (ETDEWEB)

    Palestro, C.J.; Vega, A.; Kim, C.K.; Swyer, A.J.; Goldsmith, S.J. (Mt. Sinai Medical Center, New York, NY (USA))

    1990-05-01

    Indium-111-labeled leukocyte scintigraphy was performed on a 66-yr-old male with polyarticular acute gouty arthritis. Images revealed intense labeled leukocyte accumulation in a pattern indistinguishable from septic arthritis, in both knees and ankles, and the metatarsophalangeal joint of both great toes, all of which were involved in the acute gouty attack. Joint aspirate as well as blood cultures were reported as no growth; the patient was treated with intravenous colchicine and ACTH for 10 days with dramatic improvement noted. Labeled leukocyte imaging, repeated 12 days after the initial study, revealed near total resolution of joint abnormalities, concordant with the patient's clinical improvement. This case demonstrates that while acute gouty arthritis is a potential pitfall in labeled leukocyte imaging, in the presence of known gout, it may provide a simple, objective, noninvasive method of evaluating patient response to therapy.

  9. The role of human leukocyte antigen in susceptibility and clinical manifestations of sarcoidosis.

    Institute of Scientific and Technical Information of China (English)

    1997-01-01

    To investigate the association of human leukocyte antigen (HLA) with susceptibility and clinical manifestations of sarcoidosis, fifty-five patients with sarcoidosis were studied by using allele group specific polymerase chain reaction technique (PCR). Our data

  10. Transepithelial activation of human leukocytes by probiotics and commensal bacteria: Role of Enterobacteriaceae-type endotoxin

    DEFF Research Database (Denmark)

    Baeuerlein, Annette; Ackermann, Stefanie; Parlesak, Alexandr

    2009-01-01

    The goal of the current study was to clarify whether commercially available probiotics induce greater trans-epithelial activation of human leukocytes than do commensal, food-derived and pathogenic bacteria and to identify the compounds responsible for this activation. Eleven different bacterial...... Escherichia coli K12, probiotic E. coli Nissle, EPEC) induced basolateral production of TNF-alpha, IFN-gamma, IL 6, 8, and 10. Gram-positive probiotics (Lactobacillus spp. and Bifidobacterium spp.) had virtually no effect. In addition, commensals (Enterococcus faecalis, Bacteroides vulgatus) and food...... (polymyxin, colistin) completely abrogated transepithelial activation of leukocytes. Enterobacteriaceae-type endotoxin is a crucial factor in transepithelial stimulation of leukocytes, regardless of whether it is produced by probiotics or other bacteria. Hence, transepithelial stimulation of leukocytes...

  11. The antioxidant effect of free bilirubin on cumene-hydroperoxide treated human leukocytes.

    Science.gov (United States)

    Yesilkaya, A; Altinayak, R; Korgun, D K

    2000-07-01

    To examine the antioxidant effect of bilirubin (BR) on leukocyte, we treated leukocytes obtained from healthy subjects with an oxidant and various concentrations of BR. High concentrations of BR decreased thiobarbituric acid reactive substances (TBARS) and catalase activities, increased superoxide dismutase (SOD) activity, but had no effect on glutathione (GSH) concentration. Our results showed that under physiological conditions, BR has an antioxidant effect only in high concentrations.

  12. In vivo imaging and quantitative analysis of leukocyte directional migration and polarization in inflamed tissue.

    Directory of Open Access Journals (Sweden)

    Alexander Georg Khandoga

    Full Text Available Directional migration of transmigrated leukocytes to the site of injury is a central event in the inflammatory response. Here, we present an in vivo chemotaxis assay enabling the visualization and quantitative analysis of subtype-specific directional motility and polarization of leukocytes in their natural 3D microenvironment. Our technique comprises the combination of i semi-automated in situ microinjection of chemoattractants or bacteria as local chemotactic stimulus, ii in vivo near-infrared reflected-light oblique transillumination (RLOT microscopy for the visualization of leukocyte motility and morphology, and iii in vivo fluorescence microscopy for the visualization of different leukocyte subpopulations or fluorescence-labeled bacteria. Leukocyte motility parameters are quantified off-line in digitized video sequences using computer-assisted single cell tracking. Here, we show that perivenular microinjection of chemoattractants [macrophage inflammatory protein-1alpha (MIP-1alpha/Ccl3, platelet-activating factor (PAF] or E. coli into the murine cremaster muscle induces target-oriented intravascular adhesion and transmigration as well as polarization and directional interstitial migration of leukocytes towards the locally administered stimuli. Moreover, we describe a crucial role of Rho kinase for the regulation of directional motility and polarization of transmigrated leukocytes in vivo. Finally, combining in vivo RLOT and fluorescence microscopy in Cx3CR1(gfp/gfp mice (mice exhibiting green fluorescent protein-labeled monocytes, we are able to demonstrate differences in the migratory behavior of monocytes and neutrophils.Taken together, we propose a novel approach for investigating the mechanisms and spatiotemporal dynamics of subtype-specific motility and polarization of leukocytes during their directional interstitial migration in vivo.

  13. Therapeutic relevance of penicillin-induced hypersensitivity of Staphylococcus aureus to killing by polymorphonuclear leukocytes.

    OpenAIRE

    Lam, C; Georgopoulos, A.; Laber, G.; Schütze, E

    1984-01-01

    There is an overwhelming body of evidence that certain Staphylococcus aureus strains become more sensitive to killing by polymorphonuclear leukocytes after their growth in media containing subinhibitory concentrations of penicillin. However, it is not clear to what extent this phenomenon contributes to the curative effect of penicillin in vivo. To explore its therapeutic relevance, we evaluated the interaction of staphylococci pretreated with penicillin in vitro with leukocytes in cell-proof ...

  14. Identifying the rules of engagement enabling leukocyte rolling, activation, and adhesion.

    Science.gov (United States)

    Tang, Jonathan; Hunt, C Anthony

    2010-02-19

    The LFA-1 integrin plays a pivotal role in sustained leukocyte adhesion to the endothelial surface, which is a precondition for leukocyte recruitment into inflammation sites. Strong correlative evidence implicates LFA-1 clustering as being essential for sustained adhesion, and it may also facilitate rebinding events with its ligand ICAM-1. We cannot challenge those hypotheses directly because it is infeasible to measure either process during leukocyte adhesion following rolling. The alternative approach undertaken was to challenge the hypothesized mechanisms by experimenting on validated, working counterparts: simulations in which diffusible, LFA1 objects on the surfaces of quasi-autonomous leukocytes interact with simulated, diffusible, ICAM1 objects on endothelial surfaces during simulated adhesion following rolling. We used object-oriented, agent-based methods to build and execute multi-level, multi-attribute analogues of leukocytes and endothelial surfaces. Validation was achieved across different experimental conditions, in vitro, ex vivo, and in vivo, at both the individual cell and population levels. Because those mechanisms exhibit all of the characteristics of biological mechanisms, they can stand as a concrete, working theory about detailed events occurring at the leukocyte-surface interface during leukocyte rolling and adhesion experiments. We challenged mechanistic hypotheses by conducting experiments in which the consequences of multiple mechanistic events were tracked. We quantified rebinding events between individual components under different conditions, and the role of LFA1 clustering in sustaining leukocyte-surface adhesion and in improving adhesion efficiency. Early during simulations ICAM1 rebinding (to LFA1) but not LFA1 rebinding (to ICAM1) was enhanced by clustering. Later, clustering caused both types of rebinding events to increase. We discovered that clustering was not necessary to achieve adhesion as long as LFA1 and ICAM1 object

  15. In-111 labeled leukocyte scintigraphy in a case of multifocal candidiasis

    Energy Technology Data Exchange (ETDEWEB)

    Palestro, C.J.; Vega, A.; Kim, C.K.; Goldsmith, S.J. (Mount Sinai School of Medicine, New York, NY (USA))

    1990-06-01

    The value of indium-111 labeled leukocyte scintigraphy for the diagnosis of infection in the general population is well documented; there is less information available on its role in the evaluation of the immunocompromised patient. In this study, leukocyte scintigraphy was performed on a 31-year-old immunocompromised woman who had a possible intra-abdominal abscess. No abscess was detected, but intense oral, esophageal, gastric, and vaginal uptake was observed. Candida infection was histologically confirmed at all four sites.

  16. Leukocyte extravasation as a target for anti-inflammatory therapy - Which molecule to choose?

    DEFF Research Database (Denmark)

    Boehncke, W-H; Schön, M P; Girolomoni, G

    2005-01-01

    of these agents and despite their crippling price tag, the recent incorporation of biologicals that target defined molecular controls of leukocyte extravasation into dermatological and rheumatological practise, consequently, has greatly enriched our therapeutic options for battling major, chronic, inflammatory...... dermatoses such as psoriasis. However, the - as yet unresolved and still rather controversially discussed - critical question is: Which of the multiple steps that control leukocyte extravasation in the human system really offer the most promising, most pragmatic, and safest molecular targets for therapeutic...

  17. Clinical Associations of Leukocyte Telomere Length in a Cohort of Repatriated Prisoners of War

    Science.gov (United States)

    2014-07-01

    1 RPW TELOMERE LENGTH 1. Title Page Clinical Associations of Leukocyte Telomere Length in a Cohort of Repatriated Prisoners of War Authors...Reprints will not be available from the authors Short Title: RPW Telomere Length Manuscript metrics: Word count for abstract - 200 Word count for...to 01-07-2014 4. TITLE AND SUBTITLE Clinical Associations of Leukocyte Telomere Length in a Cohort of Repatriated Prisoners of War 5a. CONTRACT

  18. Identifying the rules of engagement enabling leukocyte rolling, activation, and adhesion.

    Directory of Open Access Journals (Sweden)

    Jonathan Tang

    2010-02-01

    Full Text Available The LFA-1 integrin plays a pivotal role in sustained leukocyte adhesion to the endothelial surface, which is a precondition for leukocyte recruitment into inflammation sites. Strong correlative evidence implicates LFA-1 clustering as being essential for sustained adhesion, and it may also facilitate rebinding events with its ligand ICAM-1. We cannot challenge those hypotheses directly because it is infeasible to measure either process during leukocyte adhesion following rolling. The alternative approach undertaken was to challenge the hypothesized mechanisms by experimenting on validated, working counterparts: simulations in which diffusible, LFA1 objects on the surfaces of quasi-autonomous leukocytes interact with simulated, diffusible, ICAM1 objects on endothelial surfaces during simulated adhesion following rolling. We used object-oriented, agent-based methods to build and execute multi-level, multi-attribute analogues of leukocytes and endothelial surfaces. Validation was achieved across different experimental conditions, in vitro, ex vivo, and in vivo, at both the individual cell and population levels. Because those mechanisms exhibit all of the characteristics of biological mechanisms, they can stand as a concrete, working theory about detailed events occurring at the leukocyte-surface interface during leukocyte rolling and adhesion experiments. We challenged mechanistic hypotheses by conducting experiments in which the consequences of multiple mechanistic events were tracked. We quantified rebinding events between individual components under different conditions, and the role of LFA1 clustering in sustaining leukocyte-surface adhesion and in improving adhesion efficiency. Early during simulations ICAM1 rebinding (to LFA1 but not LFA1 rebinding (to ICAM1 was enhanced by clustering. Later, clustering caused both types of rebinding events to increase. We discovered that clustering was not necessary to achieve adhesion as long as LFA1 and

  19. Highly specific blockade of CCR5 inhibits leukocyte trafficking and reduces mucosal inflammation in murine colitis.

    Science.gov (United States)

    Mencarelli, Andrea; Cipriani, Sabrina; Francisci, Daniela; Santucci, Luca; Baldelli, Franco; Distrutti, Eleonora; Fiorucci, Stefano

    2016-08-05

    Targeted disruption of leukocyte trafficking to the gut represents a promising approach for the treatment of inflammatory bowel diseases (IBDs). CCR5, the shared receptor for MIP1α and β and RANTES, is expressed by multiple leukocytes. Here, we aimed to determine the role of CCR5 in mediating leukocyte trafficking in models of colitis, and evaluate the therapeutic potential of maraviroc, an orally active CCR5 antagonist used in the treatment of CCR5-tropic HIV. Acute and chronic colitis were induced by administration of DSS or TNBS to wild-type and CCR5(-/-) mice or adoptive transfer of splenic naïve CD4(+) T-cells from wild type or CCR5(-/-) mice into RAG-1(-/-). CCR5 gene ablation reduced the mucosal recruitment and activation of CCR5-bearing CD4(+) and CD11b(+) leukocytes, resulting in profound attenuation of signs and symptoms of inflammation in the TNBS and transfer models of colitis. In the DSS/TNBS colitis and in the transfer model, maraviroc attenuated development of intestinal inflammation by selectively reducing the recruitment of CCR5 bearing leukocytes. In summary, CCR5 regulates recruitment of blood leukocytes into the colon indicating that targeting CCR5 may offer therapeutic options in IBDs.

  20. Thrombopoietin induces p-selectin expression on platelets and subsequent platelet/leukocyte interactions.

    Science.gov (United States)

    Tibbles, Heather E; Navara, Christopher S; Hupke, Michael A; Vassilev, Alexei O; Uckun, Fatih M

    2002-04-12

    Ligation of thrombopoietin (TPO) to the platelet c-Mpl receptor induces numerous biochemical pathways in the absence of aggregation. Two forms of recombinant TPO are currently in clinical trials for the treatment of thrombocytopenia. This study focuses on the effects of the full-length recombinant human TPO (rhTPO) on platelets in a whole blood system. Platelet-leukocyte associations (PLAs) were visualized following rhTPO stimulation as CD42b/CD 45 double positive clusters by FACS analysis. Treatment of washed platelets with rhTPO induced granule release and expression of the leukocyte adhesion receptor P-selectin (CD 62P) in the absence of aggregation and calcium mobilization. RhTPO also induced platelet-leukocyte interactions in whole blood. Following stimulation, leukocytes were recruited by platelets through P-selectin in a calcium-dependent manner. rhTPO stimulates platelet-leukocyte associations in whole blood through expression of platelet P-selectin. To our knowledge, this is the first report that identifies TPO as a promoter of platelet-leukocyte interactions.

  1. Heparan sulfate domains on cultured activated glomerular endothelial cells mediate leukocyte trafficking.

    Science.gov (United States)

    Rops, A L; van den Hoven, M J; Baselmans, M M; Lensen, J F; Wijnhoven, T J; van den Heuvel, L P; van Kuppevelt, T H; Berden, J H; van der Vlag, J

    2008-01-01

    Heparan sulfate (HS) proteoglycans by playing key roles in the leukocyte-endothelial interactions are thought to mediate inflammatory cell influx in proliferative glomerulonephritis. Here, we evaluated the specific features within glomerular endothelial HS that promote leukocyte adhesion. Mouse and human glomerular endothelial cells activated by tumor necrosis factor (TNF)-alpha or interleukin (IL)-1beta increased expression of inflammatory N- and 6-O-sulfated HS domains. In addition, altered expression of HS-modifying enzymes occurred, a feature also found in mouse kidneys with anti-glomerular basement membrane disease or lupus nephritis. Inhibition of the nuclear factor (NF)-kappaB pathway repressed cytokine-induced alterations in HS and gene expression of modifying enzymes. Firm adhesion of leukocytes to activated mouse glomerular endothelial cells decreased after removal of endothelial HS or addition of sulfated heparinoids. Specific antibodies that block N- and 6-O-sulfated HS domains on activated mouse endothelial cells reduced the number of rolling and firmly adhering leukocytes under dynamic flow conditions, while they increased the average leukocyte-rolling velocity. Our study shows that N- and 6-O-sulfated domains in HS on activated glomerular endothelium are crucial for leukocyte trafficking and are possible therapeutic targets.

  2. Stress-induced enhancement of leukocyte trafficking into sites of surgery or immune activation

    Science.gov (United States)

    Viswanathan, Kavitha; Dhabhar, Firdaus S.

    2005-04-01

    Effective immunoprotection requires rapid recruitment of leukocytes into sites of surgery, wounding, infection, or vaccination. In contrast to immunosuppressive chronic stressors, short-term acute stressors have immunoenhancing effects. Here, we quantify leukocyte infiltration within a surgical sponge to elucidate the kinetics, magnitude, subpopulation, and chemoattractant specificity of an acute stress-induced increase in leukocyte trafficking to a site of immune activation. Mice acutely stressed before sponge implantation showed 200-300% higher neutrophil, macrophage, natural killer cell, and T cell infiltration than did nonstressed animals. We also quantified the effects of acute stress on lymphotactin- (LTN; a predominantly lymphocyte-specific chemokine), and TNF-- (a proinflammatory cytokine) stimulated leukocyte infiltration. An additional stress-induced increase in infiltration was observed for neutrophils, in response to TNF-, macrophages, in response to TNF- and LTN, and natural killer cells and T cells in response to LTN. These results show that acute stress initially increases trafficking of all major leukocyte subpopulations to a site of immune activation. Tissue damage-, antigen-, or pathogen-driven chemoattractants subsequently determine which subpopulations are recruited more vigorously. Such stress-induced increases in leukocyte trafficking may enhance immunoprotection during surgery, vaccination, or infection, but may also exacerbate immunopathology during inflammatory (cardiovascular disease or gingivitis) or autoimmune (psoriasis, arthritis, or multiple sclerosis) diseases. chemokine | psychophysiological stress | surgical sponge | wound healing | lymphotactin

  3. Dark chocolate consumption improves leukocyte adhesion factors and vascular function in overweight men.

    Science.gov (United States)

    Esser, Diederik; Mars, Monica; Oosterink, Els; Stalmach, Angelique; Müller, Michael; Afman, Lydia A

    2014-03-01

    Flavanol-enriched chocolate consumption increases endothelium-dependent vasodilation. Most research so far has focused on flow-mediated dilation (FMD) only; the effects on other factors relevant to endothelial health, such as inflammation and leukocyte adhesion, have hardly been addressed. We investigated whether consumption of regular dark chocolate also affects other markers of endothelial health, and whether chocolate enrichment with flavanols has additional benefits. In a randomized double-blind crossover study, the effects of acute and of 4 wk daily consumption of high flavanol chocolate (HFC) and normal flavanol chocolate (NFC) on FMD, augmentation index (AIX), leukocyte count, plasma cytokines, and leukocyte cell surface molecules in overweight men (age 45-70 yr) were investigated. Sensory profiles and motivation scores to eat chocolate were also collected. Findings showed that a 4 wk chocolate intake increased FMD by 1%, which was paralleled by a decreased AIX of 1%, decreased leukocyte cell count, decreased plasma sICAM1 and sICAM3, and decreased leukocyte adhesion marker expression (Pchocolate. This study provides new insights on how chocolate affects endothelial health by demonstrating that chocolate consumption, besides improving vascular function, also lowers the adherence capacity of leukocytes in the circulation.

  4. Identification of calprotectin, a calcium binding leukocyte protein, in human dental calculus matrix.

    Science.gov (United States)

    Kido, J; Nishikawa, S; Ishida, H; Yamashita, K; Kitamura, S; Kohri, K; Nagata, T

    1997-05-01

    Calprotectin is a calcium binding protein produced by leukocytes, macrophages and epithelial cells, and its levels in several tissues increase during infections and in many inflamed areas, suggesting that it may be an indicator of inflammatory activity. Osteopontin is a prominent phosphorylated glycoprotein in bone matrix, having calcium binding capacity. Recently, it has been reported that calprotectin and osteopontin are present in urinary stones (pathological mineralized masses in the body), and that these proteins may be involved in their formation. Dental calculus formed by mineralization of dental plaque is an inflammatory factor which may contribute to periodontal disease. It contains many organic components involved in mineralization. We recently found osteopontin molecules in human dental calculus and suggested that the components of its matrix may be similar to those of urinary stones. In this study, we investigated the presence of calprotectin in human dental calculus by immunohistochemical and immunoblotting analyses using a specific antibody for calprotectin. After fixation and demineralization of dental calculi adhered to tooth roots, sections embedded in paraffin were immunoreacted with the antibody for calprotectin and positive immunostaining for calprotectin was observed. Dental calculus proteins were then extracted with EDTA and separated by electrophoresis on 15% polyacrylamide gels. By immunoblotting analysis, 3 or 4 bands were observed at 11, 14.5, 22-25, 28 or 36.5 kDa and these patterns corresponded to those of calprotectin subunits. When non-immune rabbit serum was used instead of calprotectin-specific antibody as a negative control, no immunoreactivity was observed. These findings indicate that calprotectin is associated not only with antibacterial action but also with calcium binding capacity during dental calculus formation.

  5. Screening for arthrofibrosis after anterior cruciate ligament reconstruction: analysis of association with human leukocyte antigen.

    Science.gov (United States)

    Skutek, Michael; Elsner, Holger-A; Slateva, Kalina; Mayr, Hermann-O; Weig, Thomas-G; van Griensven, Martijn; Krettek, Christian; Bosch, Ulrich

    2004-05-01

    Arthrofibrosis represents a severe complication of trauma and reconstructive joint surgery because of generalized connective tissue proliferation resulting in painful joint stiffness. It often appears stereotypical in terms of its clinical and pathologic features, comprising excess deposition of extracellular matrix proteins such as collagen type I, III, and VI and proliferation of fibroblasts. However, trauma and surgery around joints does not always lead to fibrosis, suggesting a genetic predisposition. For a number of autoimmune diseases, strong associations have been described. The objective of the study was to investigate whether an association of HLA (human leukocyte antigen) with primary arthrofibrosis exists. Retrospective cohort study. Seventeen patients with primary arthrofibrosis after autologous anterior cruciate ligament (ACL) reconstruction were identified and clinically reviewed. Blood samples were taken, and DNA was isolated by column extraction method. DNA samples were typed for the loci HLA-A, -B, -C, -DRB1, and -DQB1. Results were compared with the frequencies of allelic groups as determined for the caucasoid population. HLA-Cw*07 was significantly less often found in the patient group than in the general population (P =.022). The opposite effect was seen for Cw*08, which was found in 17.6% of the patient group but only in 3.8% of the reference group (P =.045). A significant difference was also seen for DQB1*06, because 23.5% of the patients but 48.6% of the reference group possessed an allelic variant of this group (P =.048). However, according to the relatively small number of patients, a statistical bias cannot be excluded. A possible link may exist between arthrofibrosis and HLA-Cw*07- and DQB1*06-negative as well as Cw*08-positive individuals. Further investigation is necesessary to confirm or vitiate the possible association. Level IV.

  6. [Assessment of the response of peripheral blood mononuclear leukocytes on Helicobacter pylori infection in children].

    Science.gov (United States)

    Czkwianianc, Elzbieta; Jarosińska, Agnieszka; Chmiela, Magdalena; Bajer, Anna; Bak-Romaniszyn, Leokadia; Płaneta-Małecka, Izabela; Rudnicka, Wiesława

    2003-01-01

    In the study the proliferative response of peripheral blood mononuclear leukocytes (PBML) from children with chronic dyspepsia (chr. d) to H.p. antigens was investigated. From 38 children aged 7-18, with chr. d., blood was collected just before upper GI endoscopy. Twenty one patients were found to be H.p. (+). PBML were used for the cultures and were stimulated with heat-killed H.p. G27 bacteria, heated and unheated glycine extract (GE) of H.p. G27 or with H.p. LPS containing Lewis X and Lewis Y determinants, in the presence or absence IL-2. The cell proliferation was estimated on the basis of [3H] - thymidine incorporation. In the cultures, the phenotype of responding cells was determined by an EIA with monoclonal antibody to human CD3, CD4 and CD8. PBML from patients H.p. (-), responded to killed H.p. bacteria and to heated GE more frequently and more intensively than PBML from the H.p.(+). IL-2 enhanced PBML response to these antigens. Unheated GE did not induce PBML proliferation even in the cultures with IL-2. LPS alone induced proliferation of PBML from 3 patients (2 H.p. - and 1 H.p.+). However, in the presence of IL-2, LPS induced proliferation of PBML from 15 patients. In the cultures of PBML stimulated with whole bacteria or heated EG, T cells dominated. In the cultures of PBML from H.p. (+) we found a higher percentage of CD8 cells in comparison with the cultures of PBML from H.p. (-). Data demonstrate a significant variation in the response of PBML from dyspeptic children to H.p. antigens.

  7. Effects of cigarette smoking and nicotine metabolite ratio on leukocyte telomere length.

    Science.gov (United States)

    Verde, Zoraida; Reinoso-Barbero, Luis; Chicharro, Luis; Garatachea, Nuria; Resano, Pilar; Sánchez-Hernández, Ignacio; Rodríguez González-Moro, Jose Miguel; Bandrés, Fernando; Santiago, Catalina; Gómez-Gallego, Félix

    2015-07-01

    Studies of the effects of smoking on leukocyte telomere length (LTL) using cigarettes smoked per day or pack years smoked (PYS) present limitations. Reported high levels of smoking may not increase toxin exposure levels proportionally. Nicotine metabolism ratio (NMR) predicts total cigarette puff volume and overall exposure based on total N-nitrosamines, is highly reproducible and independent of time since the last cigarette. We hypothesized that smokers with higher NMRs will exhibit increased total puff volume, reflecting efforts to extract more nicotine from their cigarettes and increasing toxin exposure. In addition, higher levels of smoking could cause a gross damage in LTL. The urinary cotinine, 3-OH cotinine and nicotine levels of 147 smokers were analyzed using a LC/MS system Triple-Q6410. LTL and CYP2A6 genotype was determined by PCR in blood samples. We found a significant association between NMR and CYP2A6 genotype. Reduction in LTL was seen in relation to accumulated tobacco consumption and years smoking when we adjusted for age and gender. However, there were no significant differences between NMR values and LTL. In our study the higher exposure was associated with lower number of PYS. Smokers with reduced cigarette consumption may exhibit compensatory smoking behavior that results in no reduced tobacco toxin exposure. Our results suggest that lifetime accumulated smoking exposure could cause a gross damage in LTL rather than NMR or PYS. Nevertheless, a combination of smoking topography (NMR) and consumption (PYS) measures may provide useful information about smoking effects on health outcomes. Copyright © 2015 Elsevier Inc. All rights reserved.

  8. Sociodemographic correlates and family aggregation of leukocyte telomere length in adults and children from Mesoamerica.

    Science.gov (United States)

    Flannagan, Kerry S; Jansen, Erica C; Rozek, Laura S; Rentschler, Katie M; Roman, Ana Victoria; Ramirez-Zea, Manuel; Villamor, Eduardo

    2017-05-06

    Telomere length is a biomarker of cumulative stress and inflammation related to chronic disease risk. We examined the associations of leukocyte telomere length (LTL) with sociodemographic and anthropometric variables and estimated LTL family aggregation in Central America, a region with a high burden of chronic disease where LTL has not been studied. We conducted a cross-sectional study of 174 school age children and their parents in the capital cities of Belize, Honduras, Nicaragua, Costa Rica, Panama, and the city of Tuxtla-Gutierrez in Mexico. We measured LTL by quantitative PCR in DNA extracted from whole blood. We compared the distribution of LTL by categories of sociodemographic and anthropometric characteristics using linear regression. Family aggregation was estimated with correlation coefficients and intraclass correlations. In mothers, LTL was inversely associated with age (P, trend < .0001) and positively associated with height (P = .0002). Among fathers, LTL was inversely associated with food insecurity (P, trend = .0004). In children, boys had 0.10 log units shorter LTL than girls (95% CI: -0.17, -0.03; P = .004). LTL was inversely associated with parental education (P, trend = .01) and positively associated with paternal age at birth (P, trend < .0001), maternal LTL (P, trend = .007), and paternal LTL (P, trend = .02). LTL varied significantly by country of origin among all family members. Aggregation was greatest between children and their mothers, and mostly occurred at the country, rather than family, level. LTL is associated with age and height in women; food insecurity in men; and sex, parental education, parental LTL, and paternal age at birth among children. © 2016 Wiley Periodicals, Inc.

  9. Imputing amino acid polymorphisms in human leukocyte antigens.

    Directory of Open Access Journals (Sweden)

    Xiaoming Jia

    Full Text Available DNA sequence variation within human leukocyte antigen (HLA genes mediate susceptibility to a wide range of human diseases. The complex genetic structure of the major histocompatibility complex (MHC makes it difficult, however, to collect genotyping data in large cohorts. Long-range linkage disequilibrium between HLA loci and SNP markers across the major histocompatibility complex (MHC region offers an alternative approach through imputation to interrogate HLA variation in existing GWAS data sets. Here we describe a computational strategy, SNP2HLA, to impute classical alleles and amino acid polymorphisms at class I (HLA-A, -B, -C and class II (-DPA1, -DPB1, -DQA1, -DQB1, and -DRB1 loci. To characterize performance of SNP2HLA, we constructed two European ancestry reference panels, one based on data collected in HapMap-CEPH pedigrees (90 individuals and another based on data collected by the Type 1 Diabetes Genetics Consortium (T1DGC, 5,225 individuals. We imputed HLA alleles in an independent data set from the British 1958 Birth Cohort (N = 918 with gold standard four-digit HLA types and SNPs genotyped using the Affymetrix GeneChip 500 K and Illumina Immunochip microarrays. We demonstrate that the sample size of the reference panel, rather than SNP density of the genotyping platform, is critical to achieve high imputation accuracy. Using the larger T1DGC reference panel, the average accuracy at four-digit resolution is 94.7% using the low-density Affymetrix GeneChip 500 K, and 96.7% using the high-density Illumina Immunochip. For amino acid polymorphisms within HLA genes, we achieve 98.6% and 99.3% accuracy using the Affymetrix GeneChip 500 K and Illumina Immunochip, respectively. Finally, we demonstrate how imputation and association testing at amino acid resolution can facilitate fine-mapping of primary MHC association signals, giving a specific example from type 1 diabetes.

  10. Reduced Arylsulfatase B Activity in Leukocytes from Cystic Fibrosis Patients

    Science.gov (United States)

    Sharma, Girish; Burke, Jenifer; Bhattacharyya, Sumit; Sharma, Neha; Katyal, Shivani; Park, R. Lucy; Tobacman, Joanne

    2013-01-01

    Summary The enzyme Arylsulfatase B (ARSB; N-acetylgalactosamine-4-sulfatase) removes 4-sulfate groups from chondroitin-4-sulfate and dermatan sulfate and is required for the degradation of these sulfated glycosaminoglycans (GAGs). Since these GAGs accumulate in patients with Cystic Fibrosis (CF), we investigated the activity of ARSB in leukocytes of patients with CF, to consider if reduced activity of ARSB might contribute to the pathophysiology of CF. Previous cell-based experiments had demonstrated that when the deficiency of the cystic fibrosis transmembrane regulator (CFTR) was corrected in bronchial epithelial cells, the ARSB activity increased significantly. De-identified, citrated blood samples were collected from 16 children with cystic fibrosis and 31 control subjects, seen in the Pediatric Clinic at Rush University Medical Center. Polymorphonuclear (PMN) and mononuclear cell (MC) populations were separated by density gradient, and blinded determinations of ARSB activity were performed using the exogenous substrate 4-methylumbilliferyl sulfate. Interleukin-6 was measured in the plasma samples by ELISA. ARSB activity was significantly less in the PMN and MC from the CF patients than controls (p<0.0001, unpaired t-test, two-tailed). Interleukin-6 levels in plasma were significantly greater in the CF population (p<0.001). Mean age, age range, and male:female ratio of CF patients and controls were similar, and no association of ARSB activity with age, gender, or CFTR genotype was evident. Since recombinant human ARSB is used successfully for replacement therapy in Mucopolysaccharidosis VI, it may be useful to restore ARSB activity to normal levels and increase degradation of sulfated GAGs in CF patients. PMID:22550062

  11. Altered DNA methylation in leukocytes with trisomy 21.

    Directory of Open Access Journals (Sweden)

    Kristi Kerkel

    2010-11-01

    Full Text Available The primary abnormality in Down syndrome (DS, trisomy 21, is well known; but how this chromosomal gain produces the complex DS phenotype, including immune system defects, is not well understood. We profiled DNA methylation in total peripheral blood leukocytes (PBL and T-lymphocytes from adults with DS and normal controls and found gene-specific abnormalities of CpG methylation in DS, with many of the differentially methylated genes having known or predicted roles in lymphocyte development and function. Validation of the microarray data by bisulfite sequencing and methylation-sensitive Pyrosequencing (MS-Pyroseq confirmed strong differences in methylation (p<0.0001 for each of 8 genes tested: TMEM131, TCF7, CD3Z/CD247, SH3BP2, EIF4E, PLD6, SUMO3, and CPT1B, in DS versus control PBL. In addition, we validated differential methylation of NOD2/CARD15 by bisulfite sequencing in DS versus control T-cells. The differentially methylated genes were found on various autosomes, with no enrichment on chromosome 21. Differences in methylation were generally stable in a given individual, remained significant after adjusting for age, and were not due to altered cell counts. Some but not all of the differentially methylated genes showed different mean mRNA expression in DS versus control PBL; and the altered expression of 5 of these genes, TMEM131, TCF7, CD3Z, NOD2, and NPDC1, was recapitulated by exposing normal lymphocytes to the demethylating drug 5-aza-2'deoxycytidine (5aza-dC plus mitogens. We conclude that altered gene-specific DNA methylation is a recurrent and functionally relevant downstream response to trisomy 21 in human cells.

  12. Bactericidal/permeability-increasing protein preserves leukocyte functions after major liver resection.

    Science.gov (United States)

    Wiezer, M J; Meijer, C; Sietses, C; Prins, H A; Cuesta, M A; Beelen, R H; Meijer, S; van Leeuwen, P A

    2000-08-01

    To analyze postoperative leukocyte functions in patients undergoing hemihepatectomy, and to assess the effect of treatment with the endotoxin-neutralizing agent bactericidal/permeability-increasing protein (rBPI21). Extensive liver resection is associated with a high incidence of infectious complications. Because elimination of pathogenic microorganisms occurs mainly by leukocytes, this increased rate of infections is most likely due to an impaired function of these cells. Endotoxin, translocated from the gut into the systemic circulation as a result of increased gut permeability and reduced hepatic clearance function after major liver resection, may play an important role in the impairment of posthepatectomy leukocyte function. To investigate whether hemihepatectomy results in impaired leukocyte functions and to determine the role of endotoxin in this process, leukocyte oxidative burst and leukocyte antigen expression were studied in three groups of patients: patients undergoing a hemihepatectomy and receiving rBPI21 treatment, patients undergoing hemihepatectomy and receiving placebo, and as an extra control group patients undergoing other major abdominal surgeries. Blood samples were collected before surgery, 2 hours after surgery, and at days 1, 2, 5, and 7. Phorbol myristate acetate-stimulated oxidative burst was measured using dihydrorhodamine, and leukocyte surface expression of the antigens CD11b, CD16, and CD14 was investigated by indirect immunofluorescence. Both oxidative burst and membrane surface expression were quantified by flow cytometry. An indication of the antiendotoxin effect of rBPI21 treatment was provided by assessment of plasma lipopolysaccharide binding protein (LBP) levels by enzyme-linked immunosorbent assay. The oxidative burst in the hemihepatectomized patients receiving placebo and the controls increased 2 hours after surgery, whereas it decreased in the rBPI21-treated patients, resulting in significant differences between the groups

  13. Cadmium-induced microsatellite instability in the kidneys and leukocytes of C57BL/6J mice.

    Science.gov (United States)

    Du, Xiaoyan; Lan, Tianfeng; Yuan, Bao; Chen, Jian; Hu, Jinping; Ren, Wenzhi; Chen, Zhenwen

    2015-01-01

    Cadmium is a cytotoxic, carcinogenic, and mutagenic industrial product or byproduct. The correlation between metal exposure and microsatellite instability (MSI) has been reported by several groups. In the present study, 50 C57BL/6J mice at 6 weeks of age were divided into five groups and intraperitoneally injected with 0, 0.25, 0.5, 1, or 2 mg/kg cadmium chloride quaque die alterna for 4 weeks. Then, the liver, kidney, testis, leukocytes, bone marrow, and small intestine were collected from the treated mice and weighed. Portions of these tissues were fixed for further histological analysis, and the remaining tissues were subjected to genomic DNA extraction for the analysis of a panel of 42 microsatellite markers. The liver and testis weight coefficients were significantly changed in the 1 and 2 mg/kg cadmium chloride-treated groups compared with the control group. Simultaneously, severe histopathologic changes in the liver and kidneys, along with a complete disorganization of testicular structure and obvious severe necrosis in the testes were observed in the cadmium-treated group. The cadmium accumulated in the liver and kidneys of the mice in all cadmium-treated groups; the tissue cadmium concentrations were significantly higher than those in the control group. After STR scanning, MSI was found at three loci (D15Mit5, D10Mit266, and DxMit172) in the kidneys and leukocytes of mice in the lower dose groups (0.25 and 0.5 mg/kg). In summary, we have successfully established a sub-chronic cadmium exposure model and confirmed that cadmium exposure can induce MSI in mice. We also identified two loci that could be regarded as "hotspots" of microsatellite mutation in mice.

  14. Proviral genomic sequence analysis of Chinese donkey leukocyte attenuated equine infectious anemia virus vaccine and its parental virus strain Liaoning

    Institute of Scientific and Technical Information of China (English)

    王柳; 童光志; 刘红全; 杨志彪; 仇华吉; 孔宪刚; 王玫

    2002-01-01

    Proviral DNA was extracted from donkey leukocyte infected with Chinese donkey leukocyte attenuated equine infectious anemia virus(DLA-EIAV), and peripheral blood lymphocytes(PBL) from a horse infected with the virulent EIAV strain Liaoning(EIAV L). The entire proviral DNA from both viruses was cloned and sequenced. The lengths of complete genomic sequences of DLA-EIAV and EIAV L provirus were 8266 bp and 8235 bp, respectively. Sequence comparison indicated that DLA-EIAV shares 97.0% and 97.5% in sequence homology with EIAV L and donkey-adapted EIAV(DA-EIAV), respectively. Lots of variations occurred in long terminal repeat(LTR, consisting of U3, R, U5), ORF S2, and env regions between DLA-EIAV and EIAV L. The nucleotide sequence differences of the two viruses in U3, R, U5, ORF S2, and env are 13.2%, 7.5%, 5.1%, 3.9%, and 2.7%, respectively, and predicted amino acid sequence differences in env and S2 coding regions are 4.4% and 8.8%, respectively. Six conserved regions are characterized in Gp90. There is a cis-activating GATA motif in ENH of DLA-EIAV and EIAV L. Two N-linked glycosylation sites disappeared in DLA-EIAV Gp90 in comparison with that of EIAV L. A bHLH transcription factor binding consensus sequence was found in LTR of DLA-EIAV but not in EIAV L. Furthermore, there is a mutation in the stem of DLA-EIAV TAR resulting in formation of a uridine tuber. Further study is needed to uncover the relationship between sequence changes and their biological functions of DLA-EIAV and L.

  15. Peripheral nerve extract effects on mesenchymal cells.

    Science.gov (United States)

    Dietz, F R; Mukhopadhyay, B; Becker, G; Daniels, K; Solursh, M

    1996-01-01

    Several common congenital limb disorders are characterized by normal tissue differentiation but abnormal somatic growth. These include: idiopathic clubfoot, idiopathic leg length discrepancy, hemi-atrophy and hemi-hypertrophy. Both clinical and research studies have suggested that peripheral nerves may be important in regulating somatic growth of limb tissues. To investigate the hypothesis that peripheral nerves convey trophic substances to mesenchymal tissues that are involved in the regulation of growth, we developed an in vitro assay to assess the effect of fractions of peripheral nerve on myoblast and chondroblast growth and differentiation in a mammalian (rat) system. Whole rat sciatic nerve extract was fractionated by ammonium sulfate precipitation and by affinity chromatography. Concavalin A chromatography resolved whole nerve extract into a glycoprotein and a non-glycoprotein fraction. Serial ammonium sulfate precipitation yielded three pellet fractions designated as 35%, 70%, and 100% pellets; corresponding to ammonium sulfate concentrations of 0 to 35%, 35 to 70%, and 70 to 100% saturation, respectively. Dialyzed solutions of these pellets as well as the fractions from Concavalin A chromatography were assayed for biological activity in micromass cultures of rat limb bud mesenchyme, which allowed assessment of both myoblast and chondroblast stimulation. Stimulation of protein synthesis and myoblast proliferation (as measured by MF20 staining) occurred with both 70% and 100% ammonium sulfate fractions. Stimulation of chondroblasts (as measured by the number of alcian blue staining nodules) occurred with the 35% and 100% fractions. The glycoprotein fraction from the affinity chromatography stimulated protein synthesis and myoblast proliferation and inhibited chondroblast development. Stimulation of chondroblasts was seen with the non-glycoprotein fraction. No effect on protein synthesis, myoblast proliferation or chondroblast proliferation was found in

  16. Nanowire array chips for molecular typing of rare trafficking leukocytes with application to neurodegenerative pathology

    Science.gov (United States)

    Kwak, Minsuk; Kim, Dong-Joo; Lee, Mi-Ri; Wu, Yu; Han, Lin; Lee, Sang-Kwon; Fan, Rong

    2014-05-01

    Despite the presence of the blood-brain barrier (BBB) that restricts the entry of immune cells and mediators into the central nervous system (CNS), a small number of peripheral leukocytes can traverse the BBB and infiltrate into the CNS. The cerebrospinal fluid (CSF) is one of the major routes through which trafficking leukocytes migrate into the CNS. Therefore, the number of leukocytes and their phenotypic compositions in the CSF may represent important sources to investigate immune-to-brain interactions or diagnose and monitor neurodegenerative diseases. Due to the paucity of trafficking leucocytes in the CSF, a technology capable of efficient isolation, enumeration, and molecular typing of these cells in the clinical settings has not been achieved. In this study, we report on a biofunctionalized silicon nanowire array chip for highly efficient capture and multiplexed phenotyping of rare trafficking leukocytes in small quantities (50 microliters) of clinical CSF specimens collected from neurodegenerative disease patients. The antibody coated 3D nanostructured materials exhibited vastly improved rare cell capture efficiency due to high-affinity binding and enhanced cell-substrate interactions. Moreover, our platform creates multiple cell capture interfaces, each of which can selectively isolate specific leukocyte phenotypes. A comparison with the traditional immunophenotyping using flow cytometry demonstrated that our novel silicon nanowire-based rare cell analysis platform can perform rapid detection and simultaneous molecular characterization of heterogeneous immune cells. Multiplexed molecular typing of rare leukocytes in CSF samples collected from Alzheimer's disease patients revealed the elevation of white blood cell counts and significant alterations in the distribution of major leukocyte phenotypes. Our technology represents a practical tool for potentially diagnosing and monitoring the pathogenesis of neurodegenerative diseases by allowing an effective

  17. Chemokine Ligand 20: A Signal for Leukocyte Recruitment During Human Ovulation?

    Science.gov (United States)

    Al-Alem, Linah; Puttabyatappa, Muraly; Rosewell, Kathy; Brännström, Mats; Akin, James; Boldt, Jeffrey; Muse, Ken; Curry, Thomas E

    2015-09-01

    Ovulation is one of the cornerstones of female fertility. Disruption of the ovulatory process results in infertility, which affects approximately 10% of couples. Using a unique model in which the dominant follicle is collected across the periovulatory period in women, we have identified a leukocyte chemoattractant, chemokine ligand 20 (CCL20), in the human ovary. CCL20 mRNA is massively induced after an in vivo human chorionic gonadotropin (hCG) stimulus in granulosa (>10 000-fold) and theca (>4000-fold) cells collected during the early ovulatory (12-18 h) and late ovulatory (18-34 h) periods after hCG administration. Because the LH surge sets in motion an inflammatory reaction characterized by an influx of leukocytes and CCL20 is known to recruit leukocytes in other systems, the composition of ovarian leukocytes (CD45+) containing the CCL20 receptor CCR6 was determined immediately prior to ovulation. CD45+/CCR6+ cells were primarily natural killer cells (41%) along with B cells (12%), T cells (11%), neutrophils (10%), and monocytes (9%). Importantly, exogenous CCL20 stimulated ovarian leukocyte migration 59% within 90 minutes. Due to the difficulties in obtaining human follicles, an in vitro model was developed using granulosa-lutein cells to explore CCL20 regulation. CCL20 expression increased 40-fold within 6 hours after hCG, was regulated partially by the epithelial growth factor pathway, and was positively correlated with progesterone production. These results demonstrate that hCG dramatically increases CCL20 expression in the human ovary, that ovarian leukocytes contain the CCL20 receptor, and that CCL20 stimulates leukocyte migration. Our findings raise the prospect that CCL20 may aid in the final ovulatory events and contribute to fertility in women.

  18. Relationship between total and differential leukocyte counts and isolated coronary artery ectasia.

    Science.gov (United States)

    Kocaman, Sinan Altan; Taçoy, Gülten; Sahinarslan, Asife; Cengel, Atiye

    2008-08-01

    Coronary artery ectasia (CAE) is a clinical entity characterized by localized or diffuse dilatation of more than or equal to 1.5 times that of the normal adjacent segments of vessels. Although the etiopathogenesis is not clearly understood, some studies have shown that CAE may be a form of atherosclerosis and has more potent inflammatory properties. Leukocytes have a crucial role in the development of inflammatory processes. We aimed to investigate a possible relationship between leukocytes and the coronary ectatic process without coronary artery disease (CAD) and to compare it with the inflammatory atherosclerotic process related to leukocytes. The study population consisted of 371 patients. We divided the patients into three groups: 42 patients with isolated CAE as group I, 279 patients with CAD as group II, and 50 control participants with normal coronary arteries (NCA) as group III. The counts of total leukocytes (7348+/-1898, 7569+/-1619, and 6770+/-1748 cells/mm, P=0.002), neutrophils (4260+/-2169, 4529+/-1380, and 4040+/-1649 cells/mm, P=0.037) and monocytes (630+/-216, 583+/-198, and 480+/-140 cells/mm, P<0.001) were significantly different among the CAE, CAD, and NCA groups, respectively. The CAE group also had significantly higher leukocyte and subtype counts than the nonobstructive CAD subgroup and NCA group. This study demonstrates that total and differential leukocyte counts, which play an important role in inflammation, are increased in patients with isolated CAE. In conclusion, this study's findings show that leukocytes may play an important role in the development of CAE independently of the atherosclerotic process.

  19. Altered expression of adhesion molecules on peripheral blood leukocytes in feline infectious peritonitis.

    Science.gov (United States)

    Olyslaegers, Dominique A J; Dedeurwaerder, Annelike; Desmarets, Lowiese M B; Vermeulen, Ben L; Dewerchin, Hannah L; Nauwynck, Hans J

    2013-10-25

    Feline infectious peritonitis (FIP) is a fatal, coronavirus-induced systemic disease in domestic and wild felids. The pathology associated with FIP (multifocal granulomatous vasculitis) is considered to be elicited by exaggerated activation and subsequent extravasation of leukocytes. As changes in the expression of adhesion molecules on circulating leukocytes precede their margination and emigration, we reasoned that the expression of leukocyte adhesion molecules may be altered in FIP. In present study, the expression of principal adhesion molecules involved in leukocyte transmigration (CD15s, CD11a, CD11b, CD18, CD49d, and CD54) on peripheral blood leukocytes from cats with naturally occurring FIP (n=15) and controls (n=12) was quantified by flow cytometry using a formaldehyde-based rapid leukocyte preparation technique. T- and B-lymphocytes from FIP patients exhibit higher expression of both subunits (CD11a and CD18) composing the β2 integrin lymphocyte function-associated antigen (LFA)-1. In addition, the expression of the α4 subunit (CD49d) of the β1 integrin very late antigen (VLA)-4 was elevated on B-lymphocytes from FIP patients. The expression of CD11b and CD18, that combine to form the β2 integrin macrophage-1 antigen (Mac-1), was elevated on monocytes, whereas the density of CD49d was reduced on this population in FIP. Granulocytes of FIP cats displayed an increased expression of the α chain of Mac-1 (CD11b). These observations suggest that leukocytes from FIP patients show signs of systemic activation causing them to extravasate into surrounding tissues and ultimately contribute to pyogranuloma formation seen in FIP.

  20. Cells on the run: shear-regulated integrin activation in leukocyte rolling and arrest on endothelial cells.

    Science.gov (United States)

    Alon, Ronen; Ley, Klaus

    2008-10-01

    The arrest of rolling leukocytes on various target vascular beds is mediated by specialized leukocyte integrins and their endothelial immunoglobulin superfamily (IgSF) ligands. These integrins are kept in largely inactive states and undergo in situ activation upon leukocyte-endothelial contact by both biochemical and mechanical signals from flow-derived shear forces. In vivo and in vitro studies suggest that leukocyte integrin activation involves conformational alterations through inside-out signaling followed by ligand-induced rearrangements accelerated by external forces. This activation process takes place within fractions of seconds by in situ signals transduced to the rolling leukocyte as it encounters specialized endothelial-displayed chemoattractants, collectively termed arrest chemokines. In neutrophils, selectin rolling engagements trigger intermediate affinity integrins to support reversible adhesions before chemokine-triggered arrest. Different leukocyte subsets appear to use different modalities of integrin activation during rolling and arrest at distinct endothelial sites.

  1. The Alpha-Defensin Immunoassay and Leukocyte Esterase Colorimetric Strip Test for the Diagnosis of Periprosthetic Infection: A Systematic Review and Meta-Analysis.

    Science.gov (United States)

    Wyatt, M C; Beswick, A D; Kunutsor, S K; Wilson, M J; Whitehouse, M R; Blom, A W

    2016-06-15

    Synovial biomarkers have recently been adopted as diagnostic tools for periprosthetic joint infection (PJI), but their utility is uncertain. The purpose of this systematic review and meta-analysis was to synthesize the evidence on the accuracy of the alpha-defensin immunoassay and leukocyte esterase colorimetric strip test for the diagnosis of PJI compared with the Musculoskeletal Infection Society diagnostic criteria. We performed a systematic review to identify diagnostic technique studies evaluating the accuracy of alpha-defensin or leukocyte esterase in the diagnosis of PJI. MEDLINE and Embase on Ovid, ACM, ADS, arXiv, CERN DS (Conseil Européen pour la Recherche Nucléaire Document Server), CrossRef DOI (Digital Object Identifier), DBLP (Digital Bibliography & Library Project), Espacenet, Google Scholar, Gutenberg, HighWire, IEEE Xplore (Institute of Electrical and Electronics Engineers digital library), INSPIRE, JSTOR (Journal Storage), OAlster (Open Archives Initiative Protocol for Metadata Harvesting), Open Content, Pubget, PubMed, and Web of Science were searched for appropriate studies indexed from inception until May 30, 2015, along with unpublished or gray literature. The classification of studies and data extraction were performed independently by 2 reviewers. Data extraction permitted meta-analysis of sensitivity and specificity with construction of receiver operating characteristic curves for each test. We included 11 eligible studies. The pooled diagnostic sensitivity and specificity of alpha-defensin (6 studies) for PJI were 1.00 (95% confidence interval [CI], 0.82 to 1.00) and 0.96 (95% CI, 0.89 to 0.99), respectively. The area under the curve (AUC) for alpha-defensin and PJI was 0.99 (95% CI, 0.98 to 1.00). The pooled diagnostic sensitivity and specificity of leukocyte esterase (5 studies) for PJI were 0.81 (95% CI, 0.49 to 0.95) and 0.97 (95% CI, 0.82 to 0.99), respectively. The AUC for leukocyte esterase and PJI was 0.97 (95% CI, 0.95 to 0

  2. Human Leukocyte Antigen Alleles and Cytomegalovirus Infection After Renal Transplantation

    Directory of Open Access Journals (Sweden)

    Futohi

    2015-11-01

    Full Text Available Background Several studies have been conducted on the relationship between a number of human leukocyte antigen (HLA alleles and cytomegalovirus infection (CMV, in kidney transplant recipients, after transplantation. However, only a limited number of HLAs have been investigated, so far, and the results have been contradictory. Objectives This study aimed to investigate the relationship between 59 HLA alleles and the CMV infection, in transplant recipients, after kidney transplantation. Patients and Methods This retrospective cohort study was conducted on 200 patients, receiving a kidney transplant, in Baqiyatallah Hospital, in Tehran, during 2013. Throughout a one-year follow-up of kidney transplant recipients, in case of detecting the CMV antigen in patients’ blood, at any time, they were placed in the group of patients with CMV infection, whereas, if no CMV-specific antigen was developed, over a year, patients were placed in the group of patients without CMV infection, after transplantation. This study investigated the relationship between CMV infection in kidney transplant recipients and 59 HLA alleles, including 14 HLA-A, 28 HLA-B, and 17 HLA-DRB1 cases. Results Of all participants, 104 patients (52% were diagnosed with CMV infection. There was no significant difference between the two groups, with and without CMV infection, in terms of patient’s characteristics. The CMV infection, in patients receiving a transplanted organ from deceased donor, was significantly more prevalent than in those receiving kidney transplant from living donor (63% vs. 39%, respectively, P = 0.001. Recipients with HLA-B44 were more infected with CMV compared with patients without this allele (80% vs. 50%, respectively, P = 0.024; on the contrary, kidney recipients with HLA-DRB1-1 were less infected with CMV than patients without this allele (31% vs. 55%, respectively, P = 0.020. There was no significant relationship between CMV infection and other HLA alleles

  3. Microscopic observation of leukocyte kinesis in the vascular bed during hemodialysis using the rabbit ear chamber technique.

    Science.gov (United States)

    Teraoka, S; Sugawara, M; Kitano, Y; Hoshino, T; Takahashi, M; Minagawa, Y; Naganuma, S; Sanaka, T; Mineshima, M; Era, K

    1989-04-01

    Leukocyte kinesis in the capillary vascular bed during hemodialysis (HD) was investigated to elucidate the mechanism of transient leukopenia. Leukocyte movement was observed microscopically during HD using the rabbit ear chamber (REC) technique, which permits visualization of the movement of blood corpuscles in capillaries. Blood was drawn from the femoral artery and returned into the auricular and/or carotid artery so that the blood passing through the hollow fiber artificial kidney (HFAK) flowed into capillaries in the REC. Leukocyte counts of blood samples taken from the afferent and efferent limbs of the HD circuit, the right jugular vein and the right atrium were determined consecutively during HD. The difference in the leukocyte count was observed between the afferent and efferent limbs for the first 15 minutes and thereafter between the efferent limb and the jugular vein. The "transpulmonary" difference in the leukocyte count was not noticed throughout HD. Between 15 and 90 minutes after the start of HD, scarcely any circulating leukocytes were found in capillaries in the REC and some leukocytes were attached to the endothelial surface. Thereafter circulating leukocytes were seen again and detachment of leukocytes from the endothelial surface was observed. No leukocyte aggregation or embolization of aggregating leukocytes was noticed. This evidence suggests that leukopenia may be attributed to the transient shift of leukocytes to the marginal pool of the vessel lumen and this process may not be specific for the pulmonary vasculature, but may occur in the first capillary bed into which the blood passing through the HFAK flows.(ABSTRACT TRUNCATED AT 250 WORDS)

  4. Tooth extraction

    Science.gov (United States)

    ... this page: //medlineplus.gov/ency/article/007630.htm Tooth extraction To use the sharing features on this page, please enable JavaScript. A tooth extraction is a procedure to remove a tooth from ...

  5. The constitutive equation for membrane tether extraction.

    Science.gov (United States)

    Chen, Yong; Yao, Da-Kang; Shao, Jin-Yu

    2010-12-01

    Membrane tethers or nanotubes play a critical role in a variety of cellular and subcellular processes such as leukocyte rolling and intercellular mass transport. The current constitutive equations that describe the relationship between the pulling force and the tether velocity during tether extraction have serious limitations. In this article, we propose a new phenomenological constitutive equation that captures all known characteristics of nanotube formation, including nonlinearity, nonzero threshold force, and possible negative tether velocity. We used tether extraction from endothelial cells as a prototype to illustrate how to obtain the material constants in the constitutive equation. With the micropipette aspiration technique, we measured tether pulling forces at both positive and negative tether velocities. We also determined the threshold force of 55 pN experimentally for the first time. This new constitutive equation unites two established ones and provides us a unified platform to better understand not only the physiological role of tether extraction during leukocyte rolling and intercellular or intracellular transport, but also the physics of membrane tether growth or retraction.

  6. Effect of Leukocytes Transfer on the Induction of Liver Damage after Renal Ischemia- Reperfusion in Inbred Mice

    Directory of Open Access Journals (Sweden)

    Hossein Khastar

    2012-07-01

    Full Text Available Introduction: Renal ischemia-reperfusion (IR induces organ damage in remote organs such as liver, brain and lung. The aim of this study was to assess the role of leukocytes in the induction of liver damage after renal IR injury.Methods: Inbred mice were subjected to either sham operation or bilateral renal IR injury (60 min ischemia followed by 3h reperfusion. Mice were then anesthetized for collection of leukocytes by heart puncture. Isolated leukocytes were transferred to two other groups: intact recipient mice that received leukocytes from IR mice and intact recipient mice that received leukocytes from sham-operated control mice. After 24h, recipient mice were anesthetized and blood and hepatic samples were collected.Results: Alanine aminotransferase (ALT, aspartate aminotransferase (AST and hepatic malondialdehyde (MDA increased significantly in intact recipient mice that received leukocytes from IR mice in comparison to intact recipient mice receiving leukocytes from sham-operated control mice. In addition, loss of normal liver architecture, cytoplasmic vacuolization and focal infiltration of leukocytes were observed.Conclusion: These results suggest that leukocytes are one of the possible factors that contribute to liver damage after renal IR injury and this damage is partly due to the induction of oxidative stress.

  7. Directional migration of leukocytes: their pathological roles in inflammation and strategies for development of anti-inflammatory therapies

    Institute of Scientific and Technical Information of China (English)

    2001-01-01

    Directional migration of leukocytes is indispensable to innate immunity for host defense.However,recruitment of leukocytes to a site of tissue injury also constitutes a leading cause for inflammatory responses.Mechanistically,it involves a cascade of cellular events precisely regulated by temporal and spatial presentation of a repertoire of molecules in the migrating leukocytes and their surroundings(microenvironments).Here I will summarize the emerging evidence that has shed lights on the underlying molecular mechanism for directional migration of leukocytes,which has guided the therapeutical development for innovative anti-inflammatory medicines.

  8. Channel catfish (Ictalurus punctatus) leukocytes express estrogen receptor isoforms ERα and ERβ2 and are functionally modulated by estrogens

    Science.gov (United States)

    Iwanowicz, Luke R.; Stafford, James L.; Patiño, Reynaldo; Bengten, Eva; Miller, Norman W.; Blazer, Vicki

    2014-01-01

    Estrogens are recognized as modulators of immune responses in mammals and teleosts. While it is known that the effects of estrogens are mediated via leukocyte-specific estrogen receptors (ERs) in humans and mice, leucocyte-specific estrogen receptor expression and the effects of estrogens on this cell population is less explored and poorly understood in teleosts. Here in, we verify that channel catfish (Ictalurus punctaus) leukocytes express ERα and ERβ2. Transcripts of these isoforms were detected in tissue-associated leukocyte populations by PCR, but ERβ2 was rarely detected in PBLs. Expression of these receptors was temporally regulated in PBLs following polyclonal activation by concanavalin A, lipopolysaccharide or alloantigen based on evaluation by quantitative and end-point PCR. Examination of long-term leukocyte cell lines demonstrated that these receptors are differentially expressed depending on leukocyte lineage and phenotype. Expression of ERs was also temporally dynamic in some leukocyte lineages and may reflect stage of cell maturity. Estrogens affect the responsiveness of channel catfish peripheral blood leukocytes (PBLs) to mitogens in vitro. Similarly, bactericidal activity and phorbol 12-myristate 13-acetate induced respiratory burst was modulated by 17β-estradiol. These actions were blocked by the pure ER antagonist ICI 182780 indicating that response is, in part, mediated via ERα. In summary, estrogen receptors are expressed in channel catfish leukocytes and participate in the regulation of the immune response. This is the first time leukocyte lineage expression has been reported in teleost cell lines.

  9. Leukocyte telomere length and hippocampus volume: a meta-analysis [version 1; referees: 2 approved

    Directory of Open Access Journals (Sweden)

    Gustav Nilsonne

    2015-10-01

    Full Text Available Leukocyte telomere length has been shown to correlate to hippocampus volume, but effect estimates differ in magnitude and are not uniformly positive. This study aimed primarily to investigate the relationship between leukocyte telomere length and hippocampus gray matter volume by meta-analysis and secondarily to investigate possible effect moderators. Five studies were included with a total of 2107 participants, of which 1960 were contributed by one single influential study. A random-effects meta-analysis estimated the effect to r = 0.12 [95% CI -0.13, 0.37] in the presence of heterogeneity and a subjectively estimated moderate to high risk of bias. There was no evidence that apolipoprotein E (APOE genotype was an effect moderator, nor that the ratio of leukocyte telomerase activity to telomere length was a better predictor than leukocyte telomere length for hippocampus volume. This meta-analysis, while not proving a positive relationship, also is not able to disprove the earlier finding of a positive correlation in the one large study included in analyses. We propose that a relationship between leukocyte telomere length and hippocamus volume may be mediated by transmigrating monocytes which differentiate into microglia in the brain parenchyma.

  10. PLATELET-LEUKOCYTE INTERACTIONS : MULTIPLE LINKS BETWEEN INFLAMMATION , BLOOD COAGULATION AND VASCULAR RISK

    Directory of Open Access Journals (Sweden)

    Chiara Cerletti

    2010-08-01

    Full Text Available The aim of this review is to summarize the contribution of platelets and leukocytes and their interactions in inflammation and blood coagulation and its possible relevance in the pathogenesis of  thrombosis. There is some evidence of an association between infection/inflammation and thrombosis. This is likely a bidirectional relationship. The presence of a thrombus may serve as a nidus of infection. Vascular injury indeed promotes platelet and leukocyte activation and thrombus formation and the thrombus and its components facilitate adherence of bacteria to the vessel wall. Alternatively, an infection and the associated inflammation can trigger platelet and leukocyte activation and thrombus formation. In either case platelets and leukocytes co-localize and interact in the area of vascular injury, at sites of inflammation and/or at sites of thrombosis. Following vascular injury, the subendothelial tissue, a thrombogenic surface, becomes available for interaction with these blood cells. Tissue factor, found not only in media and adventitia of the vascular wall, but also on activated platelets and leukocytes, triggers blood coagulation. Vascular-blood cell interactions, mediated by the release of preformed components of the endothelium, is modulated by both cell adhesion and production of soluble stimulatory or inhibitory molecules that alter cell function: adhesion molecules regulate cell-cell contact and facilitate the modulation of biochemical pathways relevant to inflammatory and/or thrombotic processes.

  11. Exposure to Sodium Fluoride Produces Signs of Apoptosis in Rat Leukocytes

    Directory of Open Access Journals (Sweden)

    Sigrit Suástegui-Domínguez

    2010-09-01

    Full Text Available Fluoride is naturally present in the earth's crust and can be found in rocks, coal, and clay; thus, it can be found in small quantities in water, air, plants, and animals. Therefore, humans are exposed to fluoride through food, drinking water, and in the air they breathe. Flouride is essential to maintain bone strength and to protect against dental decay, but if it is absorbed too frequently, it can cause tooth decay, osteoporosis, and damage to kidneys, bones, nerves, and muscles. Therefore, the present work was aimed at determining the effect of intake of sodium fluoride (NaF as an apoptosis inducer in leukocytes of rats treated for eight weeks with 1 or 50 parts per million (ppm NaF. Expression of p53, bcl-2, and caspade-3 were used as apoptotic and general metabolism indicators of leukocyte-like indicators of the (INT oxidation system. Male rats were exposed to NaF (1 and 500 ppm for eight weeks, and then sacrificed weekly to obtain blood samples. Expression of p53, bcl-2, and caspase-3 were determined in leukocytes by Western blot, and general metabolism of leukocytes was analyzed with a commercial kit. We found changes in the expression of the proteins described, especially when the animals received 50 ppm of NaF. These results indicate that NaF intoxication can be an apoptosis inducer in rat leukocytes treated with the compound for eight weeks.

  12. Cellular softening mediates leukocyte demargination and trafficking, thereby increasing clinical blood counts.

    Science.gov (United States)

    Fay, Meredith E; Myers, David R; Kumar, Amit; Turbyfield, Cory T; Byler, Rebecca; Crawford, Kaci; Mannino, Robert G; Laohapant, Alvin; Tyburski, Erika A; Sakurai, Yumiko; Rosenbluth, Michael J; Switz, Neil A; Sulchek, Todd A; Graham, Michael D; Lam, Wilbur A

    2016-02-23

    Leukocytes normally marginate toward the vascular wall in large vessels and within the microvasculature. Reversal of this process, leukocyte demargination, leads to substantial increases in the clinical white blood cell and granulocyte count and is a well-documented effect of glucocorticoid and catecholamine hormones, although the underlying mechanisms remain unclear. Here we show that alterations in granulocyte mechanical properties are the driving force behind glucocorticoid- and catecholamine-induced demargination. First, we found that the proportions of granulocytes from healthy human subjects that traversed and demarginated from microfluidic models of capillary beds and veins, respectively, increased after the subjects ingested glucocorticoids. Also, we show that glucocorticoid and catecholamine exposure reorganizes cellular cortical actin, significantly reducing granulocyte stiffness, as measured with atomic force microscopy. Furthermore, using simple kinetic theory computational modeling, we found that this reduction in stiffness alone is sufficient to cause granulocyte demargination. Taken together, our findings reveal a biomechanical answer to an old hematologic question regarding how glucocorticoids and catecholamines cause leukocyte demargination. In addition, in a broader sense, we have discovered a temporally and energetically efficient mechanism in which the innate immune system can simply alter leukocyte stiffness to fine tune margination/demargination and therefore leukocyte trafficking in general. These observations have broad clinically relevant implications for the inflammatory process overall as well as hematopoietic stem cell mobilization and homing.

  13. Exposure to sodium fluoride produces signs of apoptosis in rat leukocytes.

    Science.gov (United States)

    Gutiérrez-Salinas, José; Morales-González, José A; Madrigal-Santillán, Eduardo; Esquivel-Soto, Jaime; Esquivel-Chirino, César; González-Rubio, Manuel García-Luna Y; Suástegui-Domínguez, Sigrit; Valadez-Vega, Carmen

    2010-09-27

    Fluoride is naturally present in the earth's crust and can be found in rocks, coal, and clay; thus, it can be found in small quantities in water, air, plants, and animals. Therefore, humans are exposed to fluoride through food, drinking water, and in the air they breathe. Flouride is essential to maintain bone strength and to protect against dental decay, but if it is absorbed too frequently, it can cause tooth decay, osteoporosis, and damage to kidneys, bones, nerves, and muscles. Therefore, the present work was aimed at determining the effect of intake of sodium fluoride (NaF) as an apoptosis inducer in leukocytes of rats treated for eight weeks with 1 or 50 parts per million (ppm) NaF. Expression of p53, bcl-2, and caspade-3 were used as apoptotic and general metabolism indicators of leukocyte-like indicators of the (INT) oxidation system. Male rats were exposed to NaF (1 and 500 ppm) for eight weeks, and then sacrificed weekly to obtain blood samples. Expression of p53, bcl-2, and caspase-3 were determined in leukocytes by Western blot, and general metabolism of leukocytes was analyzed with a commercial kit. We found changes in the expression of the proteins described, especially when the animals received 50 ppm of NaF. These results indicate that NaF intoxication can be an apoptosis inducer in rat leukocytes treated with the compound for eight weeks.

  14. KCNE gene expression is dependent on the proliferation and mode of activation of leukocytes.

    Science.gov (United States)

    Solé, Laura; Vallejo-Gracia, Albert; Roig, Sara R; Serrano-Albarrás, Antonio; Marruecos, Laura; Manils, Joan; Gómez, Diana; Soler, Concepció; Felipe, Antonio

    2013-01-01

    Voltage-dependent K (+) (Kv) channels are tightly regulated during the immune system response. Leukocytes have a limited repertoire of Kv channels, whose physiological role is under intense investigation. A functional Kv channel is an oligomeric complex composed of pore-forming and ancillary subunits. The KCNE gene family is a novel group of modulatory Kv channel elements in leukocytes. Here, we characterized the gene expression of KCNEs (1-5) in leukocytes and investigated their regulation during leukocyte proliferation and mode of activation. Murine bone-marrow-derived macrophages, human Jurkat T-lymphocytes and human Raji B-cells were analyzed. KCNEs (1-5) are expressed in all leukocytes lineages. Most KCNE mRNAs show cell cycle-dependent regulation and are differentially regulated under specific insults. Our results further suggest a new and yet undefined physiological role for KCNE subunits in the immune system. Putative associations of these ancillary proteins with Kv channels would yield a wide variety of biophysically and pharmacologically distinct channels that fine-tune the immunological response.

  15. Reduced platelet-mediated and enhanced leukocyte-mediated fibrinolysis in experimentally induced diabetes in rats

    Energy Technology Data Exchange (ETDEWEB)

    Winocour, P.D.; Colwell, J.A.

    1985-05-01

    Studies of fibrinolytic activity in diabetes mellitus have produced conflicting results. This may be a result of methodologic insensitivity or of variable contributions of the different blood components to whole blood fibrinolysis. To explore these two possibilities, the authors used a sensitive solid-phase radiometric assay to examine the fibrinolytic activity of whole blood, platelet-rich plasma, leukocytes, and platelet- and leukocyte-poor plasma prepared from control rats and rats with streptozocin-induced diabetes at various times after induction of diabetes. Fibrinolytic activity of whole blood from diabetic rats after 7 days was significantly reduced, and remained reduced after longer durations of diabetes up to 28 days. Platelet-rich plasma from diabetic rats had decreased fibrinolytic activity, which followed the same time course of changes as in whole blood. The platelet contribution to whole blood fibrinolysis was further reduced in vivo after 14 days of diabetes by a reduced whole blood platelet count. In contrast, fibrinolytic activity of leukocytes from diabetic rats became enhanced after 7 days of diabetes. After 49 days of diabetes, the whole blood leukocyte count was reduced, and in vivo would offset the enhanced activity. Plasma fibrinolytic activity was small compared with that of whole blood and was unaltered in diabetic rats. The authors conclude that altered platelet function contributes to decreased fibrinolytic activity of whole blood in diabetic rats, and that this may be partially offset by enhanced leukocyte-mediated fibrinolysis.

  16. Clinical evaluation of {sup 99m}Tc-HMPAO labeled leukocyte imaging in ulcerative colitis

    Energy Technology Data Exchange (ETDEWEB)

    Saitoh, Yasuhiro; Aburano, Tamio; Takashio, Tetsuya; Shuke, Noriyuki; Ayabe, Tokiyoshi; Nomura, Masashi; Kohgo, Yutaka; Ishikawa, Yukio; Satoh, Junichi [Asahikawa Medical Coll., Hokkaido (Japan)

    1996-07-01

    Inflammatory imaging using {sup 99m}Tc-HMPAO-labeled mixed leukocytes was assessed for use in treating 11 cases diagnosed as ulcerative colitis: 10 cases with total colitis and 1 with left-sided colitis. They consisted of 8 patients with relapse-remitting type and 3 with chronic continuous type. Radionuclide abdominal images were obtained at 1 hr, 4 hr and 24 hr after intravenous injection of 200 MBq prepared {sup 99m}Tc leukocytes. Obvious colonic activity noted at 4 hr served as the basis for positive comparative criterion in the present study. The diagnostic efficacy of radionuclide imaging was compared with endoscopic findings (based on Matts` classification) and the clinical manifestations as reference. The sensitivity and specificity of this imaging were 83.3% and 85.7%, respectively, these values being consistent with endoscopic findings and clinical manifestations at sites of disease activity. All of positive images changed to negative after treatment by leukocyte apheresis or glucocorticoid. Based on these results, {sup 99m}Tc leukocyte imaging can be used to accurately evaluate severity and treatment response in ulcerative colitis. Leukocytes may be closely related to the pathogenesis of ulcerative colitis. (author)

  17. Microfluidic chambers for monitoring leukocyte trafficking and humanized nano-proresolving medicines interactions.

    Science.gov (United States)

    Jones, Caroline N; Dalli, Jesmond; Dimisko, Laurie; Wong, Elisabeth; Serhan, Charles N; Irimia, Daniel

    2012-12-11

    Leukocyte trafficking plays a critical role in determining the progress and resolution of inflammation. Although significant progress has been made in understanding the role of leukocyte activation in inflammation, dissecting the interactions between different leukocyte subpopulations during trafficking is hampered by the complexity of in vivo conditions and the lack of detail of current in vitro assays. To measure the effects of the interactions between neutrophils and monocytes migrating in response to various chemoattractants, at single-cell resolution, we developed a microfluidic platform that replicates critical features of focal inflammation sites. We integrated an elastase assay into the focal chemotactic chambers (FCCs) of our device that enabled us to distinguish between phlogistic and nonphlogistic cell recruitment. We found that lipoxin A(4) and resolvin D1, in solution or incorporated into nano-proresolving medicines, reduced neutrophil and monocyte trafficking toward leukotriene B(4). Lipoxin A(4) also reduced the elastase release from homogenous and heterogenous mixtures of neutrophils and monocytes. Surprisingly, the effect of resolvin D1 on heterogenous mixtures was antisynergistic, resulting in a transient spike in elastase activity, which was quickly terminated, and the degraded elastin removed by the leukocytes inside the FCCs. Therefore, the microfluidic assay provides a robust platform for measuring the effect of leukocyte interactions during trafficking and for characterizing the effects of inflammation mediators.

  18. Progesterone and estrogen influence postexercise leukocyte infiltration in overiectomized female rats.

    Science.gov (United States)

    Iqbal, Sobia; Thomas, Amy; Bunyan, Kareem; Tiidus, Peter M

    2008-12-01

    Limited research has been conducted on the effects of progesterone alone, or in combination with estrogen, on leukocyte infiltration in skeletal muscle following exercise. To investigate the effects of these female sex hormones, ovariectomized female rats were divided into 4 exercise and 4 control groups: sham, estrogen, progesterone, and a combination of estrogen plus progesterone. Following 8 days of hormone replacement and 24 h postexercise, soleus (red) and superficial (white) vastus muscles were removed and immunostained for His48 (neutrophil)- and ED1 (macrophage)-positive cells. The postexercise increase in leukocyte infiltration was completely (p estrogen supplementation alone in both muscle types, relative to sham. Progesterone treatment alone also resulted in a smaller (20%-30%) but significant (p estrogen and progesterone treatment did not significantly alter the attenuation seen with estrogen supplementation alone. Hence, progesterone can independently attenuate postexercise muscle leukocyte infiltration, albeit to a lesser degree than estrogen, and it will not negate or accentuate the effect of estrogen.

  19. Leukocyte Beta-Catenin Expression Is Disturbed in Systemic Lupus Erythematosus.

    Science.gov (United States)

    Orme, Jacob J; Du, Yong; Vanarsa, Kamala; Wu, Tianfu; Satterthwaite, Anne B; Mohan, Chandra

    2016-01-01

    Wnt/β-catenin signaling is relatively understudied in immunity and autoimmunity. β-catenin blocks inflammatory mediators and favors tolerogenic dendritic cell (DC) phenotypes. We show here that leukocytes from lupus-prone mice and SLE patients express diminished β-catenin transcriptional activity, particularly in myeloid cells, although other leukocytes revealed similar trends. Serum levels of DKK-1, an inhibitor under transcriptional control of Wnt/β-catenin, were also decreased in lupus-prone mice. Surprisingly, however, preemptive deletion of β-catenin from macrophages appears to have no effect on lupus development, even in mice with varying genetic loads for lupus. Although myeloid-specific loss of β-catenin does not seem to be important for lupus development, the potential role of this transcription factor in other leukocytes and renal cells remain to be elucidated.

  20. Jon Van Rood: pioneer at the crossroad of human leukocyte antigens and transplantation.

    Science.gov (United States)

    Jansen, Jan

    2007-04-01

    Jon Van Rood (born in 1926) has made major contributions to the fields of transfusion medicine as well as organ and stem cell transplantation. His group was the first to start unraveling the complexity of the human leukocyte antigen (HLA) system through collaborative studies that used panels of sera and leukocyte samples. Furthermore, using HLA typing, he introduced the first HLA-matched platelet transfusions and developed routine leukocyte depletion as a means to prevent HLA alloimmunization. Van Rood has also been active in the fields of kidney transplantation (Eurotransplant) and stem cell transplantation (Europdonor). He combined scientific laboratory research with application to clinical medicine. He retired from his university position in 1991 but remains active in the field.

  1. TNFα regulation of Fas ligand expression on the vascular endothelium modulates leukocyte extravasation

    Science.gov (United States)

    Sata, Masataka; Walsh, Kenneth

    2010-01-01

    It is generally believed that the vascular endothelium serves as an inflammatory barrier by providing a nonadherent surface to leukocytes. Here, we report that Fas ligand (FasL) is expressed on vascular endothelial cells (ECs) and that it may function to actively inhibit leukocyte extravasation. TNFα downregulates FasL expression with an accompanying decrease in EC cytotoxicity toward co-cultured Fas-bearing cells. Local administration of TNFα to arteries downregulates endothelial FasL expression and induces mononuclear cell infiltration. Constitutive FasL expression markedly attenuates TNFα-induced cell infiltration and adherent mononuclear cells undergo apoptosis under these conditions. These findings suggest that endothelial FasL expression can negatively regulate leukocyte extravasation. PMID:9546786

  2. Human leukocyte mobilization and morphology in nickel contact allergy using a skin chamber technique

    DEFF Research Database (Denmark)

    Lerche, A; Bisgaard, H; Christensen, J D

    1981-01-01

    An improved skin chamber technique has been devised and used for quantitative evaluation of the leukocyte mobilization rate (LMR). The method was applied in 10 nickel-hypersensitive patients exposed to nickel sulphate. Each patient served as his own control and for additional control purpose, 5...... healthy individuals without nickel hypersensitivity were studied. The kinetics of the mobilized leukocytes were followed over a 48-hour period. After an initial lag phase of 2-4 hours, maximum migration was observed from the 24th to the 48th hour, with a wide interindividual variability in the number...... is a valuable means for quantitative evaluation of leukocyte mobilization and morphology in skin exudates during exposure to an allergen in delayed hypersensitivity reactions....

  3. Leukocyte Subset Changes in Response to a 164-km Road Cycle Ride in a Hot Environment

    Science.gov (United States)

    LUK, HUI-YING; MCKENZIE, AMY L.; DUPLANTY, ANTHONY A.; BUDNAR, RONALD G.; LEVITT, DANIELLE; FERNANDEZ, ALEX; LEE, ELAINE C.; ARMSTRONG, LAWRENCE E.; VINGREN, JAKOB L.

    2016-01-01

    The purpose of this observational study was to determine the circulating leukocyte subset response to completing the 2013 Hotter’N Hell Hundred recreational 164-km road cycle event in a hot and humid environmental condition. Twenty-eight men and four women were included in this study. Whole blood samples were obtained 1–2 hours before (PRE) and immediately after (POST) the event. Electronic sizing/sorting and cytometry were used to determine complete blood counts (CBC) including neutrophil, monocyte, and lymphocyte subsets. The concentration of circulating total leukocytes (103·μL−1) increased 134% from PRE to POST with the greatest increase in neutrophils (319%, pexercise in stressful environmental conditions affects the complement of circulating immune cells, although activational state and characterization of specific leukocyte subsets remains unclear. The observed increase in circulating cell sub-populations suggests that the circulating immune surveillance system may be acutely affected by exercise in hot and humid conditions. PMID:27293505

  4. Leukocyte adhesion deficiency type III: clinical features and treatment with stem cell transplantation.

    Science.gov (United States)

    Stepensky, Polina Y; Wolach, Baruch; Gavrieli, Ronit; Rousso, Sharon; Ben Ami, Tal; Goldman, Vladimir; Rozovsky, Katya; Hanna, Suhair; Etzioni, Amos; Weintraub, Michael

    2015-05-01

    Leukocyte adhesion deficiency type III (LADIII) is an autosomal recessive disorder that presents with a severe leukocyte adhesion defect and a Glanzmann-type thrombocytopathy. Hematopoietic stem cell transplantation (HSCT)--the only definitive treatment for LADIII--appears to have a high rate of complications. In this study, we describe a new group of patients with LADIII, highlighting further clinical and immunologic aspects of this disease, and reevaluating the effectiveness of HSCT for its treatment. The patients had clinical and laboratory findings consistent with LADIII. Molecular analysis confirmed the presence of a mutation in the kindlin-3 gene. HSCT was carried out in 3 patients and was successful in 2. The diagnosis of LADIII should be considered in all patients who present with recurrent infections and a bleeding diathesis, regardless of the leukocyte count. LADIII is a primary immune deficiency, which can be successfully corrected by bone marrow transplantation if applied early in the course of the disease using appropriate conditioning.

  5. [The effect of norfloxacin on the luminol-dependent chemiluminescence and adhesion of leukocytes].

    Science.gov (United States)

    Iordanova, A I; Smolkina, T V; Nikitin, A V

    1995-05-01

    The in vitro effect of norfloxacin on the luminol-dependent chemiluminescence and adherence of polymorphonuclear leukocytes of guinea pigs was studied with the use of a wide range of the drug concentrations from 0.1 to 100 micrograms/ml under two different conditions, i.e. during the cell incubation in the presence of norfloxacin various concentrations and after the cell washing to remove the drug. In a concentration of 1 microgram/ml norfloxacin stimulated the zymosan-induced chemiluminescence. When the drug was used in a concentration of 10 micrograms/ml, the stimulating effect was less pronounced. In a concentration of 100 mu/ml norfloxacin significantly inhibited the leukocyte chemiluminescence. In concentrations of 0.1 to 100 micrograms/ml norfloxacin had no effect on the leukocyte adherence to the plastic surface covered by albumin.

  6. Peripheral Leukocyte Apoptosis in Patients with Parkinsonism: Correlation with Clinical Characteristics and Neuroimaging Findings

    Directory of Open Access Journals (Sweden)

    Wei-Che Lin

    2014-01-01

    Full Text Available Apoptosis of both brain neurons and peripheral blood leukocyte is believed to be an important biomarker for evaluating the functional status of Parkinson’s disease (PD. However, their correlation remains unknown. A better understanding of the pathophysiology of neurodegeneration is essential for the treatment and prevention of PD. The present study demonstrated that leukocyte apoptosis is significantly higher in PD patients and is associated with central dopamine neuron loss by using Tc99m-TRODAT-1 SPECT. The leukocyte apoptosis and striatal dopamine transporter uptake ratios were further associated with increased severity and longer duration of disease. The interaction between brain and systemic inflammation may be responsible for the neurodegenerative disease progression.

  7. Limitations of indium-111 leukocyte scanning in febrile renal transplant patients

    Energy Technology Data Exchange (ETDEWEB)

    Sebrechts, C.; Biberstein, M.; Klein, J.L.; Witztum, K.F.

    1986-04-01

    Indium-111-labeled leukocyte scanning was evaluated as a technique for investigating possible abscess as the cause of fever in 10 renal allograft recipients under therapy for rejection, acute tubular necrosis, or urinary infection. The usefulness of the method in this setting was found to be limited by marked nonspecificity of renal, pulmonary, and other focal leukocyte accumulation. Although wound infections were correctly identified, false-positive scans resulted in multiple nonproductive consultations and radiologic procedures (some invasive) and contributed to the decision to perform one negative exploratory laparotomy. Such generalized nonspecificity in this patient population is in distinct contrast to the experience with this diagnostic test in nontransplant patients, and has not previously been reported. Possible explanations and implications of these findings are discussed. Consequently, great caution is recommended in the use of indium-111 leukocyte scans to diagnose infection in febrile renal transplant patients who present in a similar clinical setting.

  8. Fast and Specific Assessment of the Halogenating Peroxidase Activity in Leukocyte-enriched Blood Samples.

    Science.gov (United States)

    Flemmig, Jörg; Schwarz, Pauline; Bäcker, Ingo; Leichsenring, Anna; Lange, Franziska; Arnhold, Jürgen

    2016-07-28

    In this paper a protocol for the quick and standardized enrichment of leukocytes from small whole blood samples is described. This procedure is based on the hypotonic lysis of erythrocytes and can be applied to human samples as well as to blood of non-human origin. The small initial sample volume of about 50 to 100 µl makes this method applicable to recurrent blood sampling from small laboratory animals. Moreover, leukocyte enrichment is achieved within minutes and with low material efforts regarding chemicals and instrumentation, making this method applicable in multiple laboratory environments. Standardized purification of leukocytes is combined with a highly selective staining method to evaluate halogenating peroxidase activity of the heme peroxidases, myeloperoxidase (MPO) and eosinophil peroxidase (EPO), i.e., the formation of hypochlorous and hypobromous acid (HOCl and HOBr). While MPO is strongly expressed in neutrophils, the most abundant immune cell type in human blood as well as in monocytes, the related enzyme EPO is exclusively expressed in eosinophils. The halogenating activity of these enzymes is addressed by using the almost HOCl- and HOBr-specific dye aminophenyl fluorescein (APF) and the primary peroxidase substrate hydrogen peroxide. Upon subsequent flow cytometry analysis all peroxidase-positive cells (neutrophils, monocytes, eosinophils) are distinguishable and their halogenating peroxidase activity can be quantified. Since APF staining may be combined with the application of cell surface markers, this protocol can be extended to specifically address leukocyte sub-fractions. The method is applicable to detect HOCl and HOBr production both in human and in rodent leukocytes. Given the widely and diversely discussed immunological role of these enzymatic products in chronic inflammatory diseases, this protocol may contribute to a better understanding of the immunological relevance of leukocyte-derived heme peroxidases.

  9. Initial afferent lymphatic vessels controlling outbound leukocyte traffic from skin to lymph nodes.

    Directory of Open Access Journals (Sweden)

    Ignacio eMelero

    2013-12-01

    Full Text Available Tissue drains fluid and macromolecules through lymphatic vessels, which are lined by a specialized endothelium that expresses peculiar differentiation proteins, not found in blood vessels (i.e: LYVE-1, Podoplanin, PROX-1 and VEGFR-3. Lymphatic capillaries are characteristically devoid of a continuous basal membrane and are anchored to the ECM by elastic fibers that act as pulling ropes which open the vessel to avoid oedema if tissue volume increases, as it occurs upon inflammation. Lymphatic vessels are also crucial for the transit of T lymphocytes and antigen presenting cells from tissue to draining lymph nodes. Importantly, cell traffic control across lymphatic endothelium is differently regulated under resting and inflammatory conditions. Under steady-state non-inflammatory conditions, leukocytes enter into the lymphatic capillaries through basal membrane gaps (portals. This entrance is integrin-independent and seems to be mainly guided by CCL21 chemokine gradients acting on leukocytes expressing CCR7. In contrast, inflammatory processes in lymphatic capillaries involve a plethora of cytokines, chemokines, leukocyte integrins and other adhesion molecules. Importantly, under inflammation a role for integrins and their ligands becomes apparent and, as a consequence, the number of leukocytes entering the lymphatic capillaries multiplies several-fold. Enhancing transmigration of dendritic cells en route to lymph nodes is conceivably useful for vaccination and cancer immunotherapy, whereas interference with such key mechanisms may ameliorate autoimmunity or excessive inflammation. Recent findings illustrate how, transient cell-to-cell interactions between lymphatic endothelial cells and leukocytes contribute to shape the subsequent behaviour of leukocytes and condition the lymphatic vessel for subsequent trans-migratory events.

  10. Increased recruitment but impaired function of leukocytes during inflammation in mouse models of type 1 and type 2 diabetes.

    Directory of Open Access Journals (Sweden)

    Ulrika Sofia Pettersson

    Full Text Available BACKGROUND: Patients suffering from diabetes show defective bacterial clearance. This study investigates the effects of elevated plasma glucose levels during diabetes on leukocyte recruitment and function in established models of inflammation. METHODOLOGY/PRINCIPAL FINDINGS: Diabetes was induced in C57Bl/6 mice by intravenous alloxan (causing severe hyperglycemia, or by high fat diet (moderate hyperglycemia. Leukocyte recruitment was studied in anaesthetized mice using intravital microscopy of exposed cremaster muscles, where numbers of rolling, adherent and emigrated leukocytes were quantified before and during exposure to the inflammatory chemokine MIP-2 (0.5 nM. During basal conditions, prior to addition of chemokine, the adherent and emigrated leukocytes were increased in both alloxan- (62±18% and 85±21%, respectively and high fat diet-induced (77±25% and 86±17%, respectively diabetes compared to control mice. MIP-2 induced leukocyte emigration in all groups, albeit significantly more cells emigrated in alloxan-treated mice (15.3±1.0 compared to control (8.0±1.1 mice. Bacterial clearance was followed for 10 days after subcutaneous injection of bioluminescent S. aureus using non-invasive IVIS imaging, and the inflammatory response was assessed by Myeloperoxidase-ELISA and confocal imaging. The phagocytic ability of leukocytes was assessed using LPS-coated fluorescent beads and flow cytometry. Despite efficient leukocyte recruitment, alloxan-treated mice demonstrated an impaired ability to clear bacterial infection, which we found correlated to a 50% decreased phagocytic ability of leukocytes in diabetic mice. CONCLUSIONS/SIGNIFICANCE: These results indicate that reduced ability to clear bacterial infections observed during experimentally induced diabetes is not due to reduced leukocyte recruitment since sustained hyperglycemia results in increased levels of adherent and emigrated leukocytes in mouse models of type 1 and type 2 diabetes

  11. Importance of Primary Capture and L-Selectin–Dependent Secondary Capture in Leukocyte Accumulation in Inflammation and Atherosclerosis in Vivo

    Science.gov (United States)

    Eriksson, Einar E.; Xie, Xun; Werr, Joachim; Thoren, Peter; Lindbom, Lennart

    2001-01-01

    In the multistep process of leukocyte extravasation, the mechanisms by which leukocytes establish the initial contact with the endothelium are unclear. In parallel, there is a controversy regarding the role for L-selectin in leukocyte recruitment. Here, using intravital microscopy in the mouse, we investigated leukocyte capture from the free flow directly to the endothelium (primary capture), and capture mediated through interactions with rolling leukocytes (secondary capture) in venules, in cytokine-stimulated arterial vessels, and on atherosclerotic lesions in the aorta. Capture was more prominent in arterial vessels compared with venules. In venules, the incidence of capture increased with increasing vessel diameter and wall shear rate. Secondary capture required a minimum rolling leukocyte flux and contributed by ∼20–50% of total capture in all studied vessel types. In arteries, secondary capture induced formation of clusters and strings of rolling leukocytes. Function inhibition of L-selectin blocked secondary capture and thereby decreased the flux of rolling leukocytes in arterial vessels and in large (>45 μm in diameter), but not small (<45 μm), venules. These findings demonstrate the importance of leukocyte capture from the free flow in vivo. The different impact of blockage of secondary capture in venules of distinct diameter range, rolling flux, and wall shear rate provides explanations for the controversy regarding the role of L-selectin in various situations of leukocyte recruitment. What is more, secondary capture occurs on atherosclerotic lesions, a fact that provides the first evidence for roles of L-selectin in leukocyte accumulation in atherogenesis. PMID:11457895

  12. Enhancement of leukocyte adhesion after percutaneous irradiation in rats with hepatocellular carcinoma

    Institute of Scientific and Technical Information of China (English)

    Sasa-Marcel Maksan; Eduard Schmidt; Eduard Ryschich; Wolfgang Harms; Jan Schmidt

    2005-01-01

    AIM: To evaluate the effects of percutaneous radiation on leukocyte-endothelium interaction (LEI) in experimental hepatocellular carcinoma (HCC).METHODS: Twelve ACI rats underwent HCC-inoculation,six of which on day 12 received low-dose external radiation and six did not. After 12 h intravital microscopy was performed.RESULTS: LEI was significantly reduced in tumor tissue.However, irradiation of liver sinusoids and tumor tissue with 6 Gy led to a significant activation of leukocyte adhesion in the tumor with a marked increase of the proinfiammatory cytokine TNF-α.CONCLUSION: The findings indicate that the immunological tumor-endothelial barrier can be overcome by external irradiation.

  13. Effect of Vitamin C Administration on Leukocyte Vitamin C Level and Severity of Bronchial Asthma

    Directory of Open Access Journals (Sweden)

    Seyed Hamid Hashemi

    2012-04-01

    Full Text Available Oxidative stress mediated by reactive oxygen species is known to contribute to the inflammatory process of bronchial asthma. Reactive oxygen species are released into the bronchial tree by activated inflammatory cells. In this study, we aimed to determine the effect of vitamin C administration on leukocyte vitamin C level as well as severity of asthma. In this double blind clinical trial study we evaluated 60 patients with chronic stable asthma. The patients were divided into two groups (A and B including 30 patients in each group. Patients in these groups were matched according to their age, weight, height, gender, BMI and drug consumption. In addition to standard asthma treatment (according to stepwise therapy in 4th step of bronchial asthma in which the patients were controlled appropriately, group A received 1000 mg vitamin C daily and group B received placebo. At the baseline and after one month treatment, non-fasting blood samples were drawn for laboratory evaluations. Asthmatic patient's clinical condition was evaluated through standard pulmonary function test (PFT. The mean (±SD leukocyte vitamin C level in group A at the baseline and after one month treatment with 1000 mg/day vitamin C, were 0.0903 (±0.0787 µg/108 leukocytes and 0.1400 (±0.0953 µg/108 leukocytes respectively (P<0.05. The mean (±SD leukocyte vitamin C level in group B at the baseline and after one month administration of placebo, were 0.0867 (±0.0629 µg/108 leukocytes and 0.0805(±0.0736 µg/108 leukocytes respectively. The leukocyte vitamin C level in group A was higher than those of group B after one month treatment with vitamin C and placebo and the difference was statistically significant (P<0.05. Comparing PFT (FEV1, FVC and FEV1/FVC in group B during the study period showed a significant increase in FEV1 (P<0.05, while the other two parameters remained unchanged. In group A, who received 1000 mg/day vitamin C, none of the spirometry parameters changed after

  14. Radioresponse of peripheral blood and its modification by MPG (2-mercaptopropionylglycine) in mice leukocytes

    Energy Technology Data Exchange (ETDEWEB)

    Kumar, S.; Devi, P.U. (Rajasthan Univ., Jaipur (India). Radiation Biology Lab.)

    1983-01-01

    The protective effect of MPG on the peripheral blood leukocytes of Swiss albino mice was studied after a single whole-body exposure to 10 Gy of /sup 60/Co radiation with or without prior injection of MPG. The results indicated that the drug was not effective in modifying the radiation-induced changes in the total leukocyte, lymphocyte and neutrophil counts, whereas a significant drop in the number of degenerating cells was observed at the early postirradiation intervals in the drug-treated animals, indicating a protection against the direct cell killing effect of radiation.

  15. Cognitive Change during the Life Course and Leukocyte Telomere Length in Late Middle-Aged Men

    DEFF Research Database (Denmark)

    Rask, Lene; Bendix, Laila; Harbo, Maria

    2016-01-01

    , it is unclear whether there is an association between cognitive decline and leukocyte telomere length. Objective: To examine the association between changes in cognitive function during adult life and leukocyte telomere length after adjusting for confounding factors such as education, mental health and life......% CI −0.17 to −0.01, p = 0.02) but was non-significant after adjusting for smoking, alcohol consumption, leisure time activity, BMI, cholesterol, current cognitive function, depression and education (adjusted difference β = −0.07, 95% CI −0.16 to −0.01, p = 0.08). Conclusion and Relevance: Preclinical...

  16. Morphogen and proinflammatory cytokine release kinetics from PRGF-Endoret fibrin scaffolds: evaluation of the effect of leukocyte inclusion.

    Science.gov (United States)

    Anitua, E; Zalduendo, M M; Prado, R; Alkhraisat, M H; Orive, G

    2015-03-01

    The potential influence of leukocyte incorporation in the kinetic release of growth factors from platelet-rich plasma (PRP) may explain the conflicting efficiency of leukocyte platelet-rich plasma (L-PRP) scaffolds in tissue regeneration. To assess this hypothesis, leukocyte-free (PRGF-Endoret) and L-PRP fibrin scaffolds were prepared, and both morphogen and proinflammatory cytokine release kinetics were analyzed. Clots were incubated with culture medium to monitor protein release over 8 days. Furthermore, the different fibrin scaffolds were morphologically characterized. Results show that leukocyte-free fibrin matrices were homogenous while leukocyte-containing ones were heterogeneous, loose and cellular. Leukocyte incorporation produced a significant increase in the contents of proinflammatory cytokines interleukin (IL)-1β and IL-16 but not in the platelet-derived growth factors release (<1.5-fold). Surprisingly, the availability of vascular endothelial growth factor suffered an important decrease after 3 days of incubation in the case of L-PRP matrices. While the release of proinflammatory cytokines was almost absent or very low from PRGF-Endoret, the inclusion of leukocytes induced a major increase in these cytokines, which was characterized by the presence of a latent period. The PRGF-Endoret matrices were stable during the 8 days of incubation. The inclusion of leukocytes alters the growth factors release profile and also increased the dose of proinflammatory cytokines.

  17. Circulating Leukocyte and Carotid Atherosclerotic Plaque Telomere Length Interrelation, Association With Plaque Characteristics, and Restenosis After Endarterectomy

    NARCIS (Netherlands)

    Huzen, Jardi; Peeters, Wouter; de Boer, Rudolf A.; Moll, Frans L.; Wong, Liza S. M.; Codd, Veryan; de Kleijn, Dominique P. V.; de Smet, Bart J. G. L.; van Veldhuisen, Dirk J.; Samani, Nilesh J.; van Gilst, Wiek H.; Pasterkamp, Gerard; van der Harst, Pim

    2011-01-01

    Objective-Shorter leukocyte telomeres are associated with atherosclerosis and predict future heart disease. The goal of the present study was to determine whether leukocyte telomere length is related to atherosclerotic plaque telomere length and whether it is associated with plaque characteristics o

  18. Association of the leukocyte immunoglobulin G (Fcgamma) receptor IIIa-158V/F polymorphism with inflammatory myopathies in Dutch patients

    NARCIS (Netherlands)

    Bronner, I.M.; Hoogendijk, J.E.; de Visser, M.; van de Vlekkert, J.; Badrising, U.A.; Wintzen, A.R.; Uitdehaag, B.M.; Blokland-Fromme, M.; Leusen, J.H.W.; van der Pol, W.L.

    2009-01-01

    Leukocytes are involved in the pathogenesis of idiopathic inflammatory myopathies (IIMs). Immunoglobulin G (IgG) receptors (FcgammaR) link the specificity of IgG to the effector functions of leukocytes. Several FcgammaR subclasses display functional polymorphisms that determine in part the vigour of

  19. Effects of Citrus aurantifolia concentrated extract on the spontaneous proliferation of MDA-MB-453 and RPMI-8866 tumor cell lines.

    Science.gov (United States)

    Gharagozloo, M; Doroudchi, M; Ghaderi, A

    2002-07-01

    The in vitro effects of concentrated lime juice (CLJ) extract on the spontaneous proliferation of a human breast carcinoma cell line (MDA-MB-453) and a human lymphoblastoid B cell line (RPMI-8866) were investigated. CLJ extract was prepared by freeze-drying fresh fruit juice and dialyzing the concentrated extract against phosphate buffered saline in order to deplete low molecular weight micronutrients such as flavonoids as well as adjusting the pH of the extract to the physiological range. The effects of different concentrations of the CLJ extract on the spontaneous proliferative responses of the cell lines were determined by 3H-thymidine incorporation after 24 hrs of incubation. CLJ extract had no significant effect on MDA-MB-453 cell line, however, using the concentrations of 125, 250, and 500 microg/ml of CLJ extract a significant inhibition of the spontaneous proliferation of RPMI-8866 cell line was detected (P < 0.05). Due to the protein nature of the biologically active macromolecules of the CLJ extract (Gharagozloo and Ghaderi, 2001), it is reasonable to assume that the protein components of the CLJ extract may have anti-proliferative effects on tumor cell lines.

  20. Focal MMP-2 and MMP-9 Activity at the Blood-Brain Barrier Promotes Chemokine-Induced Leukocyte Migration

    Directory of Open Access Journals (Sweden)

    Jian Song

    2015-02-01

    Full Text Available Although chemokines are sufficient for chemotaxis of various cells, increasing evidence exists for their fine-tuning by selective proteolytic processing. Using a model of immune cell chemotaxis into the CNS (experimental autoimmune encephalomyelitis [EAE] that permits precise localization of immigrating leukocytes at the blood-brain barrier, we show that, whereas chemokines are required for leukocyte migration into the CNS, additional MMP-2/9 activities specifically at the border of the CNS parenchyma strongly enhance this transmigration process. Cytokines derived from infiltrating leukocytes regulate MMP-2/9 activity at the parenchymal border, which in turn promotes astrocyte secretion of chemokines and differentially modulates the activity of different chemokines at the CNS border, thereby promoting leukocyte migration out of the cuff. Hence, cytokines, chemokines, and cytokine-induced MMP-2/9 activity specifically at the inflammatory border collectively act to accelerate leukocyte chemotaxis across the parenchymal border.

  1. Cardiopulmonary bypass during cardiac surgery modulates systemic inflammation by affecting different steps of the leukocyte recruitment cascade.

    Directory of Open Access Journals (Sweden)

    Jan Rossaint

    Full Text Available BACKGROUND: It is known that the use of a cardiopulmonary bypass (CPB during cardiac surgery leads to leukocyte activation and may, among other causes, induce organ dysfunction due to increased leukocyte recruitment into different organs. Leukocyte extravasation occurs in a cascade-like fashion, including capturing, rolling, adhesion, and transmigration. However, the molecular mechanisms of increased leukocyte recruitment caused by CPB are not known. This clinical study was undertaken in order to investigate which steps of the leukocyte recruitment cascade are affected by the systemic inflammation during CPB. METHODS: We investigated the effects of CPB on the different steps of the leukocyte recruitment cascade in whole blood from healthy volunteers (n = 9 and patients undergoing cardiac surgery with the use of cardiopulmonary bypass (n = 7 or in off-pump coronary artery bypass-technique (OPCAB, n = 9 by using flow chamber experiments, transmigration assays, and biochemical analysis. RESULTS: CPB abrogated selectin-induced slow leukocyte rolling on E-selectin/ICAM-1 and P-selectin/ICAM-1. In contrast, chemokine-induced arrest and transmigration was significantly increased by CPB. Mechanistically, the abolishment of slow leukocyte rolling was due to disturbances in intracellular signaling with reduced phosphorylation of phospholipase C (PLC γ2, Akt, and p38 MAP kinase. Furthermore, CPB induced an elevated transmigration which was caused by upregulation of Mac-1 on neutrophils. CONCLUSION: These data suggest that CPB abrogates selectin-mediated slow leukocyte rolling by disturbing intracellular signaling, but that the clinically observed increased leukocyte recruitment caused by CPB is due to increased chemokine-induced arrest and transmigration. A better understanding of the underlying molecular mechanisms causing systemic inflammation after CPB may aid in the development of new therapeutic approaches.

  2. Serial Serum Leukocyte Apoptosis Levels as Predictors of Outcome in Acute Traumatic Brain Injury

    Directory of Open Access Journals (Sweden)

    Hung-Chen Wang

    2014-01-01

    Full Text Available Background. Apoptosis associates with secondary brain injury after traumatic brain injury (TBI. This study posits that serum leukocyte apoptosis levels in acute TBI are predictive of outcome. Methods. Two hundred and twenty-nine blood samples from 88 patients after acute TBI were obtained on admission and on Days 4 and 7. Serial apoptosis levels of different leukocyte subsets were examined in 88 TBI patients and 27 control subjects. Results. The leukocyte apoptosis was significantly higher in TBI patients than in controls. Brief unconsciousness (P=0.009, motor deficits (P≤0.001, GCS (P≤0.001, ISS (P=0.001, WBC count (P=0.015, late apoptosis in lymphocytes and monocytes on Day 1 (P=0.004 and P=0.022, resp., subdural hemorrhage on initial brain CT (P=0.002, neurosurgical intervention (P≤0.001, and acute posttraumatic seizure (P=0.046 were significant risk factors of outcome. Only motor deficits (P=0.033 and late apoptosis in monocytes on Day 1 (P=0.037 were independently associated with outcome. A cutoff value of 5.72% of late apoptosis in monocytes was associated with poor outcome in acute TBI patients. Conclusion. There are varying degrees of apoptosis in patients following TBI and in healthy individuals. Such differential expression suggests that apoptosis in different leukocyte subsets plays an important role in outcome following injury.

  3. Usefulness of Leukocyte Esterase Test Versus Rapid Strep Test for Diagnosis of Acute Strep Pharyngitis

    Directory of Open Access Journals (Sweden)

    Kumara V. Nibhanipudi MD

    2015-08-01

    Full Text Available Objective: A study to compare the usage of throat swab testing for leukocyte esterase on a test strip(urine dip stick-multi stick to rapid strep test for rapid diagnosis of Group A Beta hemolytic streptococci in cases of acute pharyngitis in children. Hypothesis: The testing of throat swab for leukocyte esterase on test strip currently used for urine testing may be used to detect throat infection and might be as useful as rapid strep. Methods: All patients who come with a complaint of sore throat and fever were examined clinically for erythema of pharynx, tonsils and also for any exudates. Informed consent was obtained from the parents and assent from the subjects. 3 swabs were taken from pharyngo-tonsillar region, testing for culture, rapid strep & Leukocyte Esterase. Results: Total number is 100. Cultures 9(+; for rapid strep== 84(- and16 (+; For LE== 80(- and 20(+ Statistics: From data configuration Rapid Strep versus LE test don’t seem to be a random (independent assignment but extremely aligned. The Statistical results show rapid and LE show very agreeable results. Calculated Value of Chi Squared Exceeds Tabulated under 1 Degree Of Freedom (P<.0.0001 reject Null Hypothesis and Conclude Alternative Conclusions: Leukocyte esterase on throat swab is as useful as rapid strep test for rapid diagnosis of strep pharyngitis on test strip currently used for urine dip stick causing acute pharyngitis in children.

  4. Potential Predictors of Poor Visual Outcome in Human Leukocyte Antigen-B27-Associated Uveitis

    NARCIS (Netherlands)

    Verhagen, Fleurieke H; Brouwer, Anna H; Kuiper, Jonas J W|info:eu-repo/dai/nl/342171070; Ossewaarde-van Norel, Jeannette; ten Dam-van Loon, Ninette H|info:eu-repo/dai/nl/304816957; de Boer, Joke H|info:eu-repo/dai/nl/140201890

    2016-01-01

    PURPOSE: To identify potential predictors of permanent vision loss in patients with human leukocyte antigen (HLA)-B27-associated uveitis in a tertiary referral center. DESIGN: Retrospective case-control study. METHODS: The charts of 212 patients (338 eyes) with HLA-B27-associated uveitis that visite

  5. Effect of cardiopulmonary bypass on leukocyte activation : changes in membrane-bound elastase on neutrophils

    NARCIS (Netherlands)

    Tang, M; Gu, YJ; Wang, WJ; Xu, YP; Chen, CZ

    2004-01-01

    Background: Neutrophil elastase is known to be released from the activated leukocytes as a result of cardiopulmonary bypass (CPB). However, its biological effect on organ injury is questionable because it is quickly bound by natural proteinase inhibitors (PIs). Recently, membrane-bound elastase ( MB

  6. Inhaled Nitric Oxide Decreases Leukocyte Trafficking in the Neonatal Mouse Lung During Exposure to >95% Oxygen

    Science.gov (United States)

    Rose, Melissa J.; Stenger, Michael R.; Joshi, Mandar S.; Welty, Stephen E.; Bauer, John Anthony; Nelin, Leif D.

    2010-01-01

    Chronic lung injury in the neonate is termed bronchopulmonary dysplasia (BPD). These patients generally require supplemental oxygen therapy, and hyperoxia has been implicated in the pathogenesis of BPD. The concomitant use of oxygen and inhaled nitric oxide (iNO) may result in the generation of reactive nitrogen species, or may have an anti-inflammatory effect in the neonatal lung. We tested the hypothesis that exposure to >95% O2 in neonatal mice would increase trafficking of leukocytes into the lung, and that the addition of iNO to >95% O2 would decrease this leukocyte trafficking. Hyperoxia resulted in fewer alveoli, increased presence of neutrophils and macrophages, and decreased number of mast cells within the lung parenchyma. Adding iNO to hyperoxia prevented the hyperoxia-induced changes and resulted in the numbers of alveoli, neutrophils, macrophages, and mast cells approximating those found in controls (room air exposure). Intercellular adhesion molecule (ICAM) and monocyte chemotactic protein-1 (MCP-1), two factors responsible for leukocyte recruitment, were upregulated by hyperoxic exposure, but the addition of iNO to the hyperoxic exposure prevented the hyperoxia-induced upregulation of ICAM and MCP-1. These data demonstrate that iNO alters the hyperoxia-induced recruitment of leukocytes into the lung. PMID:19915514

  7. Reduction in mitochondrial DNA copy number in peripheral leukocytes after onset of Huntington's disease

    DEFF Research Database (Denmark)

    Petersen, Maria Hvidberg; Budtz-Jørgensen, Esben; Sørensen, Sven Asger;

    2014-01-01

    to the investigation of the mitochondrial DNA (mtDNA) copy number relative to nuclear DNA (nDNA) in leukocytes from carriers of the HD mutation compared to healthy individuals. We found significantly reduced mtDNA/nDNA in HD mutation carriers compared to controls. A longitudinal study of archive DNA sample pairs from...

  8. Effects of indomethacin on ovarian leukocytes during the periovulatory period in the rat

    Directory of Open Access Journals (Sweden)

    Tarradas Esteban

    2003-02-01

    Full Text Available Abstract We have investigated the effects of indomethacin (IM, a non-steroidal anti-inflammatory drug, and the role of prostaglandins on the accumulation of leukocytes in the rat ovary during the periovulatory period. Adult cycling rats were injected sc with 1 mg of IM in olive oil or vehicle on the morning of proestrus. Some animals were killed at 16:00 h in proestrus. On the evening (19:00 h of proestrus, IM-treated rats were injected with 500 micrograms of prostaglandin E1 in saline or vehicle. Animals were killed at 01:30 and 09:00 h in estrus. There was an influx of macrophages, neutrophils, and eosinophils into the theca layers of preovulatory follicles, and of neutrophils and eosinophils into the ovarian medulla from 16:00 h in proestrus to 01:30 h in estrus. All these changes, except the accumulation of neutrophils in the theca layers of preovulatory follicles, were blocked by IM treatment. At 09:00 h in estrus, large clusters of neutrophils were observed in IM-treated rats, around abnormally ruptured follicles. The accumulation of leukocytes was not restored by prostaglandin supplementation, despite the inhibition of abnormal follicle rupture and restoration of ovulation in these animals. These results suggest that different mechanisms are involved in leukocyte accumulation in the ovary during the periovulatory period, and that the inhibitory effects of IM on the influx of leukocytes are not dependent on prostaglandin synthesis inhibition.

  9. Secretory leukocyte proteinase inhibitor-competent DNA deposits are potent stimulators of plasmacytoid dendritic cells

    DEFF Research Database (Denmark)

    Skrzeczynska-Moncznik, Joanna; Wlodarczyk, Agnieszka; Zabieglo, Katarzyna

    2012-01-01

    Secretory leukocyte proteinase inhibitor (SLPI) is a well-established inhibitor of serine proteases such as human neutrophil elastase (HNE) and a NF-κB regulatory agent in immune cells. In this paper, we report that SLPI plays a previously uncharacterized role in regulating activation of plasmacy...

  10. A simple skin blister technique for the study of in vivo transmigration of human leukocytes.

    Science.gov (United States)

    Davidsson, Lisa; Björkman, Lena; Christenson, Karin; Alsterholm, Mikael; Movitz, Charlotta; Thorén, Fredrik B; Karlsson, Anna; Welin, Amanda; Bylund, Johan

    2013-07-31

    The study of human leukocytes is almost exclusively conducted using cells isolated from peripheral blood. This is especially true for neutrophils, despite the fact that these cells are of main (pathological) importance in extravascular tissues upon e.g., infection and/or tissue damage. The journey from circulation to tissue is typically associated with a number of cellular changes, making the cells primed, or hyper-responsive, and in many aspects distinct from the cells present in circulation. Models to obtain in vivo transmigrated leukocytes from human tissue are available, but not widely used. We describe here an easy-to-use model for the study of local inflammation, stemming from limited tissue damage, which can be used to isolate viable and functional leukocytes. The model is based on the generation of aseptic skin blisters, formed by the application of negative pressure, and allows for investigations of the cellular infiltrate as well as of soluble mediators present in the exudate. We believe that this method, combined with modern analysis equipment suitable for small volumes and cell numbers, could be of great use for increasing our understanding of the nature and function of leukocytes that have left circulation and transmigrated to inflamed tissues. Copyright © 2013 Elsevier B.V. All rights reserved.

  11. Association between Leukocyte Mitochondrial DNA Copy Number and Regular Exercise in Postmenopausal Women.

    Science.gov (United States)

    Chang, Yu Kyung; Kim, Da Eun; Cho, Soo Hyun; Kim, Jung-Ha

    2016-11-01

    Previous studies suggest that habitual exercise can improve skeletal mitochondrial function; however, to date, the association between exercise and mitochondrial function in peripheral leukocytes has not been reported. The aim of this study was to evaluate the relationship between regular exercise and mitochondrial function by measuring leukocyte mitochondrial DNA (mtDNA) copy number in postmenopausal women. This cross-sectional study included 144 relatively healthy, non-diabetic, non-smoking, postmenopausal women. Clinical parameters, including anthropometric measurements and cardio-metabolic parameters, were assessed. Regular exercise was defined as at least 150 minutes per week of moderate-intensity activity, or an equivalent combination of moderate and vigorous-intensity activity, over a duration of at least 6 months. Leukocyte mtDNA copy numbers were measured using real-time polymerase chain reaction assays, and these were normalized to the β-globin copy number to give the relative mtDNA copy number. The mtDNA copy number of peripheral leukocytes was significantly greater in the exercise group (1.33±0.02) than in the no exercise group (1.05±0.02, Pcopy number (β=0.25, Pcopy number in postmenopausal women.

  12. Isolation of RNA and DNA from leukocytes and cDNA synthesis.

    NARCIS (Netherlands)

    Jansen, J.H.; Reijden, B.A. van der

    2006-01-01

    In this chapter, methods to isolate RNA and DNA from human leukocytes for the subsequent use in molecular diagnostic tests are described. In addition, protocols for cDNA synthesis are given, both for the use in conventional reverse transcription (RT)-polymerase chain reaction (PCR), and for the use

  13. Human neutrophil defensins and secretory leukocyte proteinase inhibitor in squamous metaplastic epithelium of bronchial airways.

    NARCIS (Netherlands)

    Aarbiou, J.; Schadewijk, A. van; Stolk, J.; Sont, J.K.; Boer, W.I.; Rabe, K.F.; Krieken, J.H.J.M. van; Mauad, T.; Hiemstra, P.S.

    2004-01-01

    OBJECTIVE: The aim of this study was to analyze a possible contribution of human neutrophil defensins and secretory leukocyte proteinase inhibitor (SLPI) to the induction of airway epithelial changes such as squamous cell metaplasia. MATERIALS AND METHODS: The presence of these molecules and the num

  14. Phylogenetic analysis of the swine leukocyte antigen - 2 gene for Korean native pigs

    Science.gov (United States)

    The objective of this study was to investigate genetic distances of the SLA-2 gene, to characterize SLA-2 alleles, and to provide basic genetic information of Korean pigs. The swine leukocyte antigen - 2 (SLA-2) gene in the MHC classical region was cloned with spleen tissues from Korean native pigs ...

  15. Activated leukocyte cell adhesion molecule expression predicts lymph node metastasis in oral squamous cell carcinoma.

    NARCIS (Netherlands)

    Brand, M. van den; Takes, R.P.; Blokpoel-deRuyter, M.; Slootweg, P.J.; Kempen, L.C.L.T. van

    2010-01-01

    Lymphatic metastasis of oral squamous cell carcinoma (SCC) is important for prognosis and clinical decision making concerning the treatment of the neck but may be difficult to detect. Activated leukocyte cell adhesion molecule (ALCAM), has been shown to correlate with prognosis or tumor grade in dif

  16. Leukocyte oxygen radical production determines disease severity in the recurrent Guillain-Barré syndrome

    Directory of Open Access Journals (Sweden)

    Bergström Tomas

    2010-08-01

    Full Text Available Abstract Background The recurrent Guillain-Barré syndrome (RGBS is characterized by at least two GBS episodes with intervening remission. In a previous study of monophasic GBS, we reported that the magnitude of oxygen radical production ("respiratory burst" in peripheral blood leukocytes was inversely correlated to disease severity. The present study sought to establish a similar correlation in patients with RGBS. Methods Oxygen radical production in leukocytes was induced by formyl-Met-Leu-Phe (fMLF, Trp-Lys-Tyr-Met-Val-Met-NH2 (WKYMVM, or phorbol myristate acetate (PMA and assessed by quantifying superoxide anion formed by the leukocyte NADPH oxidase. Results Disease severity, assessed using the MRC score, was negatively correlated to superoxide anion production triggered by fMLF or WKYMVM (p = 0.001 and 0.002, respectively; n = 10. Superoxide anion production also was significantly lower in RGBS patients with incomplete recovery after stimulation with fMLF (p = 0.004 or WKYMVM (p = 0.003. Conclusion We conclude that a lower respiratory burst in leukocytes is strongly associated with a severe course of RGBS.

  17. Leukocyte oxygen radical production determines disease severity in the recurrent Guillain-Barré syndrome.

    Science.gov (United States)

    Mossberg, Natalia; Andersen, Oluf; Nordin, Magnus; Nilsson, Staffan; Svedhem, Ake; Bergström, Tomas; Hellstrand, Kristoffer; Movitz, Charlotta

    2010-08-08

    The recurrent Guillain-Barré syndrome (RGBS) is characterized by at least two GBS episodes with intervening remission. In a previous study of monophasic GBS, we reported that the magnitude of oxygen radical production ("respiratory burst") in peripheral blood leukocytes was inversely correlated to disease severity. The present study sought to establish a similar correlation in patients with RGBS. Oxygen radical production in leukocytes was induced by formyl-Met-Leu-Phe (fMLF), Trp-Lys-Tyr-Met-Val-Met-NH2 (WKYMVM), or phorbol myristate acetate (PMA) and assessed by quantifying superoxide anion formed by the leukocyte NADPH oxidase. Disease severity, assessed using the MRC score, was negatively correlated to superoxide anion production triggered by fMLF or WKYMVM (p = 0.001 and 0.002, respectively; n = 10). Superoxide anion production also was significantly lower in RGBS patients with incomplete recovery after stimulation with fMLF (p = 0.004) or WKYMVM (p = 0.003). We conclude that a lower respiratory burst in leukocytes is strongly associated with a severe course of RGBS.

  18. Dark chocolate consumption improves leukocyte adhesion factors and vascular function in overweight men

    NARCIS (Netherlands)

    Esser, D.; Mars, M.; Oosterink, E.; Stalmach, A.; Müller, M.R.; Afman, L.A.

    2014-01-01

    Flavanol-enriched chocolate consumption increases endothelium-dependent vasodilation. Most research so far has focused on flow-mediated dilation (FMD) only; the effects on other factors relevant to endothelial health, such as inflammation and leukocyte adhesion, have hardly been addressed. We

  19. Transfusion associated complications in cardiac surgery : the swan song of the allogeneic leukocytes ?

    NARCIS (Netherlands)

    Bilgin, Memiş Yavuz

    2011-01-01

    In the Netherlands 1 of 1.000 inhibitants undergo cardiac surgery annually. To compensate blood loss these patients receive often blood transfusions, which can cause unexpected adverse reactions. Allogeneic leukocytes may play a prominent role in the development of these adverse reactions. We found

  20. Enzyme analysis for Pompe disease in leukocytes; superior results with natural substrate compared with artificial substrates.

    Science.gov (United States)

    van Diggelen, O P; Oemardien, L F; van der Beek, N A M E; Kroos, M A; Wind, H K; Voznyi, Y V; Burke, D; Jackson, M; Winchester, B G; Reuser, A J J

    2009-06-01

    Enzyme analysis for Pompe disease in leukocytes has been greatly improved by the introduction of acarbose, a powerful inhibitor of interfering alpha-glucosidases, which are present in granulocytes but not in lymphocytes. Here we show that the application of acarbose in the enzymatic assay employing the artificial substrate 4-methylumbelliferyl-alpha-D: -glucoside (MU-alphaGlc) is insufficient to clearly distinguish patients from healthy individuals in all cases. Also, the ratios of the activities without/with acarbose only marginally discriminated Pompe patients and healthy individuals. By contrast, when the natural substrate glycogen is used, the activity in leukocytes from patients (n = 82) with Pompe disease is at most 17% of the lowest control value. The use of artificial substrate in an assay with isolated lymphocytes instead of total leukocytes is a poor alternative as blood samples older than one day invariably yield lymphocyte preparations that are contaminated with granulocytes. To diagnose Pompe disease in leukocytes we recommend the use of glycogen as substrate in the presence of acarbose. This assay unequivocally excludes Pompe disease. To also exclude pseudo-deficiency of acid alpha-glucosidase caused by the sequence change c.271G>A (p.D91N or GAA2; homozygosity in approximately 1:1000 caucasians), a second assay employing MU-alphaGlc substrate plus acarbose or DNA analysis is required.

  1. Cell-stiffness-induced mechanosignaling - a key driver of leukocyte transendothelial migration

    NARCIS (Netherlands)

    A. Schaefer; P.L. Hordijk

    2015-01-01

    The breaching of cellular and structural barriers by migrating cells is a driving factor in development, inflammation and tumor cell metastasis. One of the most extensively studied examples is the extravasation of activated leukocytes across the vascular endothelium, the inner lining of blood vessel

  2. Varicella zoster virus glycoprotein C increases chemokine-mediated leukocyte migration.

    Science.gov (United States)

    González-Motos, Víctor; Jürgens, Carina; Ritter, Birgit; Kropp, Kai A; Durán, Verónica; Larsen, Olav; Binz, Anne; Ouwendijk, Werner J D; Lenac Rovis, Tihana; Jonjic, Stipan; Verjans, Georges M G M; Sodeik, Beate; Krey, Thomas; Bauerfeind, Rudolf; Schulz, Thomas F; Kaufer, Benedikt B; Kalinke, Ulrich; Proudfoot, Amanda E I; Rosenkilde, Mette M; Viejo-Borbolla, Abel

    2017-05-01

    Varicella zoster virus (VZV) is a highly prevalent human pathogen that establishes latency in neurons of the peripheral nervous system. Primary infection causes varicella whereas reactivation results in zoster, which is often followed by chronic pain in adults. Following infection of epithelial cells in the respiratory tract, VZV spreads within the host by hijacking leukocytes, including T cells, in the tonsils and other regional lymph nodes, and modifying their activity. In spite of its importance in pathogenesis, the mechanism of dissemination remains poorly understood. Here we addressed the influence of VZV on leukocyte migration and found that the purified recombinant soluble ectodomain of VZV glycoprotein C (rSgC) binds chemokines with high affinity. Functional experiments show that VZV rSgC potentiates chemokine activity, enhancing the migration of monocyte and T cell lines and, most importantly, human tonsillar leukocytes at low chemokine concentrations. Binding and potentiation of chemokine activity occurs through the C-terminal part of gC ectodomain, containing predicted immunoglobulin-like domains. The mechanism of action of VZV rSgC requires interaction with the chemokine and signalling through the chemokine receptor. Finally, we show that VZV viral particles enhance chemokine-dependent T cell migration and that gC is partially required for this activity. We propose that VZV gC activity facilitates the recruitment and subsequent infection of leukocytes and thereby enhances VZV systemic dissemination in humans.

  3. Mechanisms involved in apoptosis of carp leukocytes upon in vitro and in vivo immunostimulation

    NARCIS (Netherlands)

    Kepka, M.; Verburg-van Kemenade, B.M.L.; Homa, J.; Chadzinska, M.K.

    2014-01-01

    During inflammation leukocyte activity must be carefully regulated, as high concentrations and/or prolonged action of pro-inflammatory mediators e.g. reactive oxygen species (ROS) can be detrimental not only for pathogens but also for host tissues. Programmed cell death – apoptosis is a most effecti

  4. Effect of Intensity of Cigarette Smoking on Leukocytes among Adult Men and Women Smokers in Bangladesh

    Directory of Open Access Journals (Sweden)

    Shahena Shipa

    2017-03-01

    Full Text Available Background: Smoking is one of the preventable causes of disease in middle and low-income countries. This study was conducted in smokers and non-smokers to observe the changes in total count of leukocytes in cigarette smokers in relation to body mass index (BMI and blood pressure (BP. Methods:The study populations were from different sources including diagnostic center and general hospital, and consisted of 58 smokers and 77 non-smokers, with a broad range of age groups. The variables considered for this study were the smoking status of current smokers and non-smokers, and blood samples of the subject, anthropometric data and also blood pressure data. Results: Total leukocytes in smokers were found to be higher than the non-smokers along with the increasing of lymphocytes. Leukocytes were also found to be increased with intensity of smoking in adult men and women. The BMI of the smokers showed decreasing trend compared to non-smokers. The relation between blood pressure and smoking was not well established, as there were only little changes on systolic blood pressure (SBP of smokers found according to smoking intensity. Conclusion: Cigarette smoking has negative effects on leukocytes both in men and women smokers in terms of certain anthropometric parameters.

  5. Leptin and Adiponectin: new players in the field of tumor cell and leukocyte migration

    Directory of Open Access Journals (Sweden)

    Ratke Janina

    2009-12-01

    Full Text Available Abstract Adipose tissue is no longer considered to be solely an energy storage, but exerts important endocrine functions, which are primarily mediated by a network of various soluble factors derived from fat cells, called adipocytokines. In addition to their responsibility to influence energy homeostasis, new studies have identified important pathways linking metabolism with the immune system, and demonstrating a modulatory role of adipocytokines in immune function. Additionally, epidemiological studies underline that obesity represents a significant risk factor for the development of cancer, although the exact mechanism of this relationship remains to be determined. Whereas a possible influence of adipocytokines on the proliferation of tumor cells is already known, new evidence has come to light elucidating a modulatory role of this signaling substances in the regulation of migration of leukocytes and tumor cells. The migration of leukocytes is a key feature to fight cancer cells, whereas the locomotion of tumor cells is a prerequisite for tumor formation and metastasis. We herein review the latest tumor biological findings on the role of the most prominent adipocytokines leptin and adiponectin, which are secreted by fat cells, and which are involved in leukocyte migration, tumor growth, invasion and metastasis. This review thus accentuates the complex, interactive involvement of adipocytokines in the regulation of migration of both leukocytes and tumor cells, and gives an insight in the underlying molecular mechanisms.

  6. In vitro leukocyte adhesion to modified polyurethane surfaces. II. Effect of wettability

    NARCIS (Netherlands)

    Bruil, Anton; Brenneisen, Laura M.; Terlingen, Johannes G.A.; Beugeling, Tom; Aken, van Willem G.; Feijen, J.

    1994-01-01

    The influence of substrate wettability on leukocyte adhesion was studied using a series of polyurethane films with different surface wettabilities, prepared by a two step gas plasma modification procedure. In the first step the films were made hydrophobic by exposure to a tetrafluoromethane plasma.

  7. Dark chocolate consumption improves leukocyte adhesion factors and vascular function in overweight men

    NARCIS (Netherlands)

    Esser, D.; Mars, M.; Oosterink, E.; Stalmach, A.; Müller, M.R.; Afman, L.A.

    2014-01-01

    Flavanol-enriched chocolate consumption increases endothelium-dependent vasodilation. Most research so far has focused on flow-mediated dilation (FMD) only; the effects on other factors relevant to endothelial health, such as inflammation and leukocyte adhesion, have hardly been addressed. We invest

  8. No Evidence of Human Leukocyte Antigen Gene Association With Rheumatic Fever Among Children in Samoa.

    Science.gov (United States)

    Erdem, Guliz; Seifried, Steven E

    2015-03-01

    Human leukocyte antigens (HLAs) have been implicated in rheumatic fever pathogenesis. This pilot whole genome association study compares genotypes of Samoan children with rheumatic fever to unaffected siblings and unrelated healthy controls. No risk-related genotypes were associated with HLA genes. Thirteen Regions of Interest were identified as candidates for further study.

  9. Recent advances in microscopic techniques for visualizing leukocytes in vivo [version 1; referees: 3 approved

    Directory of Open Access Journals (Sweden)

    Rohit Jain

    2016-05-01

    Full Text Available Leukocytes are inherently motile and interactive cells. Recent advances in intravital microscopy approaches have enabled a new vista of their behavior within intact tissues in real time. This brief review summarizes the developments enabling the tracking of immune responses in vivo.

  10. Potential Predictors of Poor Visual Outcome in Human Leukocyte Antigen-B27-Associated Uveitis

    NARCIS (Netherlands)

    Verhagen, Fleurieke H; Brouwer, Anna H; Kuiper, Jonas J W; Ossewaarde-van Norel, Jeannette; ten Dam-van Loon, Ninette H; de Boer, Joke H

    2016-01-01

    PURPOSE: To identify potential predictors of permanent vision loss in patients with human leukocyte antigen (HLA)-B27-associated uveitis in a tertiary referral center. DESIGN: Retrospective case-control study. METHODS: The charts of 212 patients (338 eyes) with HLA-B27-associated uveitis that visite

  11. Familial occurrence of subacute thyroiditis associated with human leukocyte antigen-B35

    NARCIS (Netherlands)

    Kramer, AB; Roozendaal, C; Dullaart, RPF

    2004-01-01

    Subacute thyroiditis (SAT) is a spontaneously remitting inflammatory disorder of the thyroid, associated with human leukocyte antigen (HLA)-B35, and may be virally induced in genetically predisposed individuals. A 57-year-old Caucasian man presented with symptoms of hyperthyroidism as well as enlarg

  12. Vero cytotoxin binding to polymorphonuclear leukocytes among households with children with hemolytic uremic syndrome.

    NARCIS (Netherlands)

    Loo, D.M.W.M. te; Heuvelink, A.E.; Boer, E. de; Nauta, J.; Walle, J. van der; Schröder, C.H.; Hinsbergh, V.W.H. van; Chart, H.; Kar, N.C.A.J. van de; Heuvel, L.P.W.J. van den

    2001-01-01

    Hemolytic uremic syndrome (HUS), the leading cause of acute renal failure in childhood, can be caused by different serotypes of vero cytotoxin (VT; i.e., Shiga toxin)-producing Escherichia coli (VTEC). Recently, VT was shown to bind to polymorphonuclear leukocytes (PMNL) in the systemic circulation

  13. Leukocyte integrin Mac-1 regulates thrombosis via interaction with platelet GPIbα.

    Science.gov (United States)

    Wang, Yunmei; Gao, Huiyun; Shi, Can; Erhardt, Paul W; Pavlovsky, Alexander; A Soloviev, Dmitry; Bledzka, Kamila; Ustinov, Valentin; Zhu, Liang; Qin, Jun; Munday, Adam D; Lopez, Jose; Plow, Edward; Simon, Daniel I

    2017-05-30

    Inflammation and thrombosis occur together in many diseases. The leukocyte integrin Mac-1 (also known as integrin αMβ2, or CD11b/CD18) is crucial for leukocyte recruitment to the endothelium, and Mac-1 engagement of platelet GPIbα is required for injury responses in diverse disease models. However, the role of Mac-1 in thrombosis is undefined. Here we report that mice with Mac-1 deficiency (Mac-1(-/-)) or mutation of the Mac-1-binding site for GPIbα have delayed thrombosis after carotid artery and cremaster microvascular injury without affecting parameters of haemostasis. Adoptive wild-type leukocyte transfer rescues the thrombosis defect in Mac-1(-/-) mice, and Mac-1-dependent regulation of the transcription factor Foxp1 contributes to thrombosis as evidenced by delayed thrombosis in mice with monocyte-/macrophage-specific overexpression of Foxp1. Antibody and small-molecule targeting of Mac-1:GPIbα inhibits thrombosis. Our data identify a new pathway of thrombosis involving leukocyte Mac-1 and platelet GPIbα, and suggest that targeting this interaction has anti-thrombotic therapeutic potential with reduced bleeding risk.

  14. Sphingosine-1-phosphate receptor 3 promotes leukocyte rolling by mobilizing endothelial P-selectin.

    Science.gov (United States)

    Nussbaum, Claudia; Bannenberg, Sarah; Keul, Petra; Gräler, Markus H; Gonçalves-de-Albuquerque, Cassiano F; Korhonen, Hanna; von Wnuck Lipinski, Karin; Heusch, Gerd; de Castro Faria Neto, Hugo C; Rohwedder, Ina; Göthert, Joachim R; Prasad, Vysakh Pushpa; Haufe, Günter; Lange-Sperandio, Baerbel; Offermanns, Stefan; Sperandio, Markus; Levkau, Bodo

    2015-01-01

    Sphingosine-1-phosphate (S1P) participates in inflammation; however, its role in leukocyte rolling is still unclear. Here we use intravital microscopy in inflamed mouse cremaster muscle venules and human endothelial cells to show that S1P contributes to P-selectin-dependent leukocyte rolling through endothelial S1P receptor 3 (S1P3) and Gαq, PLCβ and Ca(2+). Intra-arterial S1P administration increases leukocyte rolling, while S1P3 deficiency or inhibition dramatically reduces it. Mast cells involved in triggering rolling also release S1P that mobilizes P-selectin through S1P3. Histamine and epinephrine require S1P3 for full-scale effect accomplishing it by stimulating sphingosine kinase 1 (Sphk1). In a counter-regulatory manner, S1P1 inhibits cAMP-stimulated Sphk1 and blocks rolling as observed in endothelial-specific S1P1(-/-) mice. In agreement with a dominant pro-rolling effect of S1P3, FTY720 inhibits rolling in control and S1P1(-/-) but not in S1P3(-/-) mice. Our findings identify S1P as a direct and indirect contributor to leukocyte rolling and characterize the receptors mediating its action.

  15. Localization and activity of inflammatory bowel disease using /sup 111/In leukocyte imaging

    Energy Technology Data Exchange (ETDEWEB)

    Hotze, A.; Biersack, H.J.; Biele, B.; Wolf, F.; Knapp, F.F.

    1988-06-01

    A prospective study was initiated to compare the clinically proven results concerning localization/extent and activity of inflammatory bowel diseases with those of /sup 111/In-oxine leukocyte imaging. All patients studied were completely examined with barium enema x-ray, clinical and laboratory investigations, and endoscopy with histopathology. A total of 31 leukocyte scans were performed in 15 patients (12 with Crohn's desease, 3 with ulcerative colitis). The scans were graded by comparing the cell uptake of a lesion (when present) and a bone marrow area providing a count ratio (CR). The inflammatory lesions were correctly localized on 26 leukocyte scans, and in 21 scans the scintigraphically estimated extent of disease was identical to endoscopy. In 5 cases the disease extent was underestimated, 4 scans in patients with relapse of Crohn's disease were falsely negative, and in one patient with remission truly negative. The scintigraphically assessed disease activity was also in a good agreement with clinical disease activity based on histopathology in all cases. We conclude that leukocyte imaging provides valuable information about localization and activity of inflammatory bowel disease.

  16. Rac-null leukocytes are associated with increased inflammation-mediated alveolar bone loss.

    Science.gov (United States)

    Sima, Corneliu; Gastfreund, Shoshi; Sun, Chunxiang; Glogauer, Michael

    2014-02-01

    Periodontitis is characterized by altered host-biofilm interactions that result in irreversible inflammation-mediated alveolar bone loss. Genetic and epigenetic factors that predispose to ineffective control of biofilm composition and maintenance of tissue homeostasis are not fully understood. We elucidated how leukocytes affect the course of periodontitis in Rac-null mice. Mouse models of acute gingivitis and periodontitis were used to assess the early inflammatory response and patterns of chronicity leading to loss of alveolar bone due to inflammation in Rac-null mice. Leukocyte margination was differentially impaired in these mice during attachment in conditional Rac1-null (granulocyte/monocyte lineage) mice and during rolling and attachment in Rac2-null (all blood cells) mice. Inflammatory responses to subgingival ligatures, assessed by changes in peripheral blood differential leukocyte numbers, were altered in Rac-null compared with wild-type mice. In response to persistent subgingival ligature-mediated challenge, Rac-null mice had increased loss of alveolar bone with patterns of resorption characteristic of aggressive forms of periodontitis. These findings were partially explained by higher osteoclastic coverage of the bone-periodontal ligament interface in Rac-null compared with wild-type mice. In conclusion, this study demonstrates that leukocyte defects, such as decreased endothelial margination and tissue recruitment, are rate-limiting steps in the periodontal inflammatory process that lead to more aggressive forms of periodontitis.

  17. Rapid, high-efficiency labeling of leukocytes with In-111 after hemolytic removal of erythrocytes

    Energy Technology Data Exchange (ETDEWEB)

    Karesh, S.M.; Henkin, R.E.

    1985-05-01

    During the labeling of leukocytes with Indium-111, conventional methodology involves separation and washing to remove red cells. This technique results in the loss of a significant number of leukocytes. Citrated whole blood of ten normal volunteers was studied for an alternate labeling method following sedimentation for 30 to 45 minutes and low speed centrifugation of the leukocyte-rich plasma. The average labeling for these ten volunteers by Indium-111 was 90% versus 60% by the older technique. Viability as measured by the trypan blue exclusion test was greater than 95%, WBC losses were essentially zero, and no WBC clumping was observed. Eighteen patients referred for leukocyte imaging were studied by this method. In this patient population, there was 91% labeling with viability greater than 95% and no evidence of clumping. Less than 5% RBC's were noted in any lot. Indium-111 WBC activity 20 minutes post injection averaged 79% of whole blood activity. This modification results in decreased losses of white cells, reduces preparation time to less than 2 hours, and significantly improves the labeling efficiency of the final product. Liver/spleen ratios and image quality were unchanged from the original method.

  18. Poly(ethyleneimine) modified filters for the removal of leukocytes from blood

    NARCIS (Netherlands)

    Bruil, Anton; Oosterom, Hieke A.; Steneker, Ingeborg

    1993-01-01

    Polyurethane membrane filters and filters coated with poly(ethyleneimine) were used to investigate the influence of leukocyte adhesion during filtration. Treatment of the filters with an aqueous solution of 1% (w/v) poly(ethyleneimine) (PEI) led to the introduction of amine groups at the filter surf

  19. Transfusion of Leukocyte-Depleted RBCs Is Independently Associated With Increased Morbidity After Pediatric Cardiac Surgery

    NARCIS (Netherlands)

    Kneyber, Martin C. J.; Grotenhuis, Femke; Berger, Rolf F. M.; Ebels, Tjark W.; Burgerhof, Johannes G. M.; Albers, Marcel J. I. J.

    2013-01-01

    Objective: To test the hypothesis that transfusion of leukocyte-depleted RBC preparations within the first 48 hours of PICU stay was independently associated with prolonged duration of mechanical ventilation, irrespective of surgery type and disease severity. Design: Retrospective, observational stu

  20. Mechanisms involved in apoptosis of carp leukocytes upon in vitro and in vivo immunostimulation

    NARCIS (Netherlands)

    Kepka, M.; Verburg-van Kemenade, B.M.L.; Homa, J.; Chadzinska, M.K.

    2014-01-01

    During inflammation leukocyte activity must be carefully regulated, as high concentrations and/or prolonged action of pro-inflammatory mediators e.g. reactive oxygen species (ROS) can be detrimental not only for pathogens but also for host tissues. Programmed cell death – apoptosis is a most

  1. Reduction of leukocyte-derived H2S linked to abnormal glycolipid metabolism in hypertensive subjects.

    Science.gov (United States)

    Sun, Xiaonan; Chen, Yongzeng; Zeng, Qiang; Huang, Xianyong; Cai, Junyan

    2017-01-01

    We deduced that leukocyte-derived H2S would also play a pivotal role regarding nutrition homeostasis in hypertensive subjects. Plasma was obtained from patients with hypertension (n  =  151) as well as control (n  =  41). Leukocyte-derived H2S speed was determined, and biochemical indices of glucose and lipid metabolism were measured. Western blot analyses of CSE were also performed. Inflammation factors were measured. Leukocyte-derived H2S is produced at a significantly lower rate in overweight or obese patients (p H2S and the levels of HOMA-RI and insulin in overweight patients and has a positive relationship with HDL-C only in overweight hypertensive patients (p H2S speed (r = 0.995, p = 0.001). Linear regression analysis showed that insulin levels will increase (β = -1.685, p = 0.041) with the slower speed of H2S. Leukocyte-derived H2S production varied according to the nutritional status of hypertensive subjects, and the H2S/IL-10 signaling pathway may be the junction point among hypertension, disturbance of nutritional status, and inflammation.

  2. Tracking and fixed ranking of leukocyte telomere length across the adult life course

    DEFF Research Database (Denmark)

    Benetos, Athanase; Kark, Jeremy D; Susser, Ezra;

    2013-01-01

    Short leukocyte telomere length (LTL) is associated with atherosclerosis in adults and diminished survival in the elderly. LTL dynamics are defined by LTL at birth, which is highly variable, and its age-dependent attrition thereafter, which is rapid during the first 20 years of life. We examined...

  3. Leukocyte telomere length is inversely correlated with plasma Von Willebrand factor

    DEFF Research Database (Denmark)

    Hjelmborg, Jacob V B; Nzietchueng, Rosine; Kimura, Masayuki;

    2010-01-01

    INTRODUCTION: Leukocyte telomere length (LTL) is short, while the plasma level of Von Willebrand (VWF) is high in persons with atherosclerosis. Moreover, both short LTLs and high VWF levels are observed in individuals who display risks for atherosclerosis, including hypertension, obesity, insulin...

  4. Higher circulating levels of IGF-1 are associated with longer leukocyte telomere length in healthy subjects

    DEFF Research Database (Denmark)

    Barbieri, Michelangela; Paolisso, Giuseppe; Kimura, Masayuki;

    2009-01-01

    Mutations that inhibit the insulin-like growth factor-1 (IGF-1) extend the lifespan of worms, flies and mice. However, it appears that relatively low circulating levels of IGF-1 in humans are associated with aging-related diseases and diminished longevity. As leukocyte telomere length (LTL...

  5. Association of leukocyte telomere length with fatigue in nondisabled older adults

    DEFF Research Database (Denmark)

    Bendix, Laila; Thinggaard, Mikael; Kimura, Masayuki;

    2014-01-01

    Introduction: Fatigue is often present in older adults with no identified underlying cause. The accruing burden of oxidative stress and inflammation might be underlying factors of fatigue. We therefore hypothesized that leukocyte telomere length (LTL) is relatively short in older adults who...

  6. Leukocyte telomere length and physical ability among Danish twins age 70+

    DEFF Research Database (Denmark)

    Bendix, Laila; Gade, Maria Monrad; Staun, Pia Wirenfeldt;

    2011-01-01

    Leukocyte telomere length (LTL) shortens with age and is potentially a biomarker of human aging. We examined the relation of LTL with physical ability and cognitive function in 548 same-sex twins from the Longitudinal Study of Aging Danish Twins. LTL was measured by Southern blots of the terminal...

  7. Varicella zoster virus glycoprotein C increases chemokine-mediated leukocyte migration.

    Directory of Open Access Journals (Sweden)

    Víctor González-Motos

    2017-05-01

    Full Text Available Varicella zoster virus (VZV is a highly prevalent human pathogen that establishes latency in neurons of the peripheral nervous system. Primary infection causes varicella whereas reactivation results in zoster, which is often followed by chronic pain in adults. Following infection of epithelial cells in the respiratory tract, VZV spreads within the host by hijacking leukocytes, including T cells, in the tonsils and other regional lymph nodes, and modifying their activity. In spite of its importance in pathogenesis, the mechanism of dissemination remains poorly understood. Here we addressed the influence of VZV on leukocyte migration and found that the purified recombinant soluble ectodomain of VZV glycoprotein C (rSgC binds chemokines with high affinity. Functional experiments show that VZV rSgC potentiates chemokine activity, enhancing the migration of monocyte and T cell lines and, most importantly, human tonsillar leukocytes at low chemokine concentrations. Binding and potentiation of chemokine activity occurs through the C-terminal part of gC ectodomain, containing predicted immunoglobulin-like domains. The mechanism of action of VZV rSgC requires interaction with the chemokine and signalling through the chemokine receptor. Finally, we show that VZV viral particles enhance chemokine-dependent T cell migration and that gC is partially required for this activity. We propose that VZV gC activity facilitates the recruitment and subsequent infection of leukocytes and thereby enhances VZV systemic dissemination in humans.

  8. Use of {sup 99m}Tc-Mononuclear Leukocyte Scintigraphy in Nosocomial Fever

    Energy Technology Data Exchange (ETDEWEB)

    Gutfilen, B.; Lopes de Souza, S.A.; Martins, F.P.P.; Cardoso, L.R.; Pinheiro Pessoa, M.C.; Fonseca, L.M.B. [Univ. Federal do Rio de Janeiro (Brazil). Dept. de Radiologia

    2006-09-15

    Purpose: To determine the overall diagnostic accuracy of mononuclear leukocyte-{sup 99m}Tc scintigraphy in the routine detection of infectious lesions and fever of unknown origin (FUO) in inpatients. Material and Methods: The use of mononuclear leukocyte {sup 99m}Tc scintigraphy is presented in 87 patients who fulfilled the Durack and Street diagnostic criteria of nosocomial FUO; 66 patients were suspected of having infectious lesions (myocarditis, endocarditis, infected catheters, diabetic foot, and osteomyelitis) and 21 patients presented with unknown causes of FUO. Scans were carried out 1, 3, and 24 h after injection of labeled leukocytes. Results: In three cases (3/27) where scintigraphs were negative, biopsies were positive. There were two (2/87) false-positive scintigrams. We found a 95.8% sensitivity and 92.3% specificity. PPV was 93.8%, PPN 94.7%, and accuracy 94.2%. Conclusion: Mononuclear leukocyte {sup 99m}Tc scintigraphy showed high sensitivity, specificity, positive and negative predictive values in patients with nosocomial FUO. These results suggest an important role for nuclear medicine in the management of patients with infection/inflammation.

  9. Long telomeres in blood leukocytes are associated with a high risk of ascending aortic aneurysm.

    Directory of Open Access Journals (Sweden)

    Tuija J Huusko

    Full Text Available Ascending aortic aneurysm is a connective tissue disorder. Even though multiple novel gene mutations have been identified, risk profiling and diagnosis before rupture still represent a challenge. There are studies demonstrating shorter telomere lengths in the blood leukocytes of abdominal aortic aneurysm patients. The aim of this study was to measure whether relative telomere lengths are changed in the blood leukocytes of ascending aortic aneurysm patients. We also studied the expression of telomerase in aortic tissue samples of ascending aortic aneurysms. Relative lengths of leukocyte telomeres were determined from blood samples of patients with ascending aortic aneurysms and compared with healthy controls. Telomerase expression, both at the level of mRNA and protein, was quantified from the aortic tissue samples. Mean relative telomere length was significantly longer in ascending aortic aneurysm blood samples compared with controls (T/S ratio 0.87 vs. 0.61, p<0.001. Expressions of telomerase mRNA and protein were elevated in the aortic aneurysm samples (p<0.05 and p<0.01. Our study reveals a significant difference in the mean length of blood leukocyte telomeres in ascending aortic aneurysm and controls. Furthermore, expression of telomerase, the main compensating factor for telomere loss, is elevated at both the mRNA and protein level in the samples of aneurysmal aorta. Further studies will be needed to confirm if this change in telomere length can serve as a tool for assessing the risk of ascending aortic aneurysm.

  10. Electromembrane extraction

    DEFF Research Database (Denmark)

    Huang, Chuixiu; Chen, Zhiliang; Gjelstad, Astrid

    2017-01-01

    Electromembrane extraction (EME) was inspired by solid-phase microextraction and developed from hollow fiber liquid-phase microextraction in 2006 by applying an electric field over the supported liquid membrane (SLM). EME provides rapid extraction, efficient sample clean-up and selectivity based...

  11. Grape polyphenols and propolis mixture inhibits inflammatory mediator release from human leukocytes and reduces clinical scores in experimental arthritis.

    Science.gov (United States)

    Mossalayi, M D; Rambert, J; Renouf, E; Micouleau, M; Mérillon, J M

    2014-02-15

    Polyphenols from red fruits and bee-derived propolis (PR) are bioactive natural products in various in vitro and in vivo models. The present study shows that hematotoxicity-free doses of grape polyphenols (GPE) and PR differentially decreased the secretion of pro-inflammatory cytokines from activated human peripheral blood leucocytes. While GPE inhibited the monocytes/macrophage response, propolis decreased both monokines and interferon γ (IFNγ) production. When used together, their distinct effects lead to the attenuation of all inflammatory mediators, as supported by a significant modulation of the transcriptomic profile of pro-inflammatory genes in human leukocytes. To enforce in vitro data, GPE+PR were tested for their ability to improve clinical scores and cachexia in chronic rat adjuvant-induced arthritis (AA). Extracts significantly reduced arthritis scores and cachexia, and this effect was more significant in animals receiving continuous low doses compared to those receiving five different high doses. Animals treated daily had significantly better clinical scores than corticoid-treated rats. Together, these findings indicate that the GPE+PR combination induces potent anti-inflammatory activity due to their complementary immune cell modulation. Copyright © 2013 Elsevier GmbH. All rights reserved.

  12. A novel method for rapid and reliable detection of complex vertebral malformation and bovine leukocyte adhesion deficiency in Holstein cattle

    Directory of Open Access Journals (Sweden)

    Zhang Yi

    2012-07-01

    Full Text Available Abstract Background Complex vertebral malformation (CVM and bovine leukocyte adhesion deficiency (BLAD are two autosomal recessive lethal genetic defects frequently occurring in Holstein cattle, identifiable by single nucleotide polymorphisms. The objective of this study is to develop a rapid and reliable genotyping assay to screen the active Holstein sires and determine the carrier frequency of CVM and BLAD in Chinese dairy cattle population. Results We developed real-time PCR-based assays for discrimination of wild-type and defective alleles, so that carriers can be detected. Only one step was required after the DNA extraction from the sample and time consumption was about 2 hours. A total of 587 Chinese Holstein bulls were assayed, and fifty-six CVM-carriers and eight BLAD-carriers were identified, corresponding to heterozygote carrier frequencies of 9.54% and 1.36%, respectively. The pedigree analysis showed that most of the carriers could be traced back to the common ancestry, Osborndale Ivanhoe for BLAD and Pennstate Ivanhoe Star for CVM. Conclusions These results demonstrate that real-time PCR is a simple, rapid and reliable assay for BLAD and CVM defective allele detection. The high frequency of the CVM allele suggests that implementing a routine testing system is necessary to gradually eradicate the deleterious gene from the Chinese Holstein population.

  13. Misoprostol Inhibits Lipopolysaccharide-Induced Pro-inflammatory Cytokine Production by Equine Leukocytes

    Directory of Open Access Journals (Sweden)

    Emily Medlin Martin

    2017-09-01

    Full Text Available Pro-inflammatory cytokines including tumor necrosis factor α (TNFα, IL-1β, IL-6, and IL-8 are potent immune mediators that exacerbate multiple equine diseases such as sepsis and laminitis. Unfortunately, safe and effective cytokine-targeting therapies are lacking in horses; therefore, novel mechanisms of inhibiting cytokine production are critically needed. One potential mechanism for inhibiting cytokine synthesis is elevation of intracellular cyclic AMP (cAMP. In human leukocytes, intracellular cAMP production is induced by activation of E-prostanoid (EP receptors 2 and 4. These receptors can be targeted by the EP2/4 agonist and prostaglandin E1 analog, misoprostol. Misoprostol is currently used as a gastroprotectant in horses but has not been evaluated as a cytokine-targeting therapeutic. Thus, we hypothesized that misoprostol treatment would inhibit pro-inflammatory cytokine production by lipopolysaccharide (LPS-stimulated equine leukocytes in an in vitro inflammation model. To test this hypothesis, equine leukocyte-rich plasma (LRP was collected from 12 healthy adult horses and used to model LPS-mediated inflammatory signaling. LRP was treated with varying concentrations of misoprostol either before (pretreated or following (posttreated LPS stimulation. LRP supernatants were assayed for 23 cytokines using an equine-specific multiplex bead immunoassay. Leukocytes were isolated from LRP, and leukocyte mRNA levels of four important cytokines were evaluated via RT-PCR. Statistical differences between treatments were determined using one-way RM ANOVA (Holm–Sidak post hoc testing or Friedman’s RM ANOVA on Ranks (SNK post hoc testing, where appropriate (p < 0.05, n = 3–6 horses. These studies revealed that misoprostol pre- and posttreatment inhibited LPS-induced TNFα and IL-6 protein production in equine leukocytes but had no effect on IL-8 protein. Interestingly, misoprostol pretreatment enhanced IL-1β protein synthesis

  14. Vacuum extraction

    DEFF Research Database (Denmark)

    Maagaard, Mathilde; Oestergaard, Jeanett; Johansen, Marianne

    2012-01-01

    Objectives. To develop and validate an Objective Structured Assessment of Technical Skills (OSATS) scale for vacuum extraction. Design. Two part study design: Primarily, development of a procedure-specific checklist for vacuum extraction. Hereafter, validationof the developed OSATS scale for vacuum...... extraction in a prospective observational study. Setting. Rigshospitalet, University Hospital of Copenhagen. Population. For development an obstetric expert from each labor ward in Denmark (28 departments) were invited to participate. For validation nine first-year residents and ten chief physicians...... with daily work in the obstetric field were tested. Methods. The Delphi method was used for development of the scale. In a simulated vacuum extraction scenario first-year residents and obstetric chief physicians were rated using the developed OSATS scale for vacuum extraction to test construct validity...

  15. Vacuum extraction

    DEFF Research Database (Denmark)

    Maagaard, Mathilde; Oestergaard, Jeanett; Johansen, Marianne

    2012-01-01

    Objectives. To develop and validate an Objective Structured Assessment of Technical Skills (OSATS) scale for vacuum extraction. Design. Two part study design: Primarily, development of a procedure-specific checklist for vacuum extraction. Hereafter, validationof the developed OSATS scale for vacuum...... extraction in a prospective observational study. Setting. Rigshospitalet, University Hospital of Copenhagen. Population. For development an obstetric expert from each labor ward in Denmark (28 departments) were invited to participate. For validation nine first-year residents and ten chief physicians...... with daily work in the obstetric field were tested. Methods. The Delphi method was used for development of the scale. In a simulated vacuum extraction scenario first-year residents and obstetric chief physicians were rated using the developed OSATS scale for vacuum extraction to test construct validity...

  16. Influence of Leukocyte- and Platelet-Rich Fibrin (L-PRF in the Healing of Simple Postextraction Sockets: A Split-Mouth Study

    Directory of Open Access Journals (Sweden)

    Gaetano Marenzi

    2015-01-01

    Full Text Available The aim of this study was to evaluate the effects of leukocyte- and platelet-rich fibrin (L-PRF on the pain and soft tissue healing after tooth extractions. Twenty-six patients (9 males and 17 females were treated with multiple extractions (2 to 8, with a total of 108 extractions. This was an exploratory single blinded randomized clinical trial with a split-mouth design. The pain after the surgery was assessed in each patient by the VAS scale (1 to 10 at intervals of 24-48-72-96 hours. The soft tissue healing was clinically evaluated at 3, 7, 14, and 21 days after surgery by the same examiner surgeon, using the modified Healing Index (4 to 12. The mean value of postextraction pain was 3.2 ± 0.3 in the experimental sides and 4.1 ± 0.1 in the control sides. After 7 days from the extractions, the values of modified Healing Index in the experimental and control groups were, respectively, 4.8 ± 0.6 and 5.1 ± 0.9. The use of L-PRF in postextraction sockets filling can be proposed as a useful procedure in order to manage the postoperative pain and to promote the soft tissue healing process, reducing the early adverse effects of the inflammation.

  17. Influence of Leukocyte- and Platelet-Rich Fibrin (L-PRF) in the Healing of Simple Postextraction Sockets: A Split-Mouth Study.

    Science.gov (United States)

    Marenzi, Gaetano; Riccitiello, Francesco; Tia, Mariano; di Lauro, Alessandro; Sammartino, Gilberto

    2015-01-01

    The aim of this study was to evaluate the effects of leukocyte- and platelet-rich fibrin (L-PRF) on the pain and soft tissue healing after tooth extractions. Twenty-six patients (9 males and 17 females) were treated with multiple extractions (2 to 8), with a total of 108 extractions. This was an exploratory single blinded randomized clinical trial with a split-mouth design. The pain after the surgery was assessed in each patient by the VAS scale (1 to 10) at intervals of 24-48-72-96 hours. The soft tissue healing was clinically evaluated at 3, 7, 14, and 21 days after surgery by the same examiner surgeon, using the modified Healing Index (4 to 12). The mean value of postextraction pain was 3.2 ± 0.3 in the experimental sides and 4.1 ± 0.1 in the control sides. After 7 days from the extractions, the values of modified Healing Index in the experimental and control groups were, respectively, 4.8 ± 0.6 and 5.1 ± 0.9. The use of L-PRF in postextraction sockets filling can be proposed as a useful procedure in order to manage the postoperative pain and to promote the soft tissue healing process, reducing the early adverse effects of the inflammation.

  18. Roscovitine blocks leukocyte extravasation by inhibition of cyclin-dependent kinases 5 and 9.

    Science.gov (United States)

    Berberich, Nina; Uhl, Bernd; Joore, Jos; Schmerwitz, Ulrike K; Mayer, Bettina A; Reichel, Christoph A; Krombach, Fritz; Zahler, Stefan; Vollmar, Angelika M; Fürst, Robert

    2011-07-01

    Roscovitine, a cyclin-dependent kinase (CDK) inhibitor that induces tumour cell death, is under evaluation as an anti-cancer drug. By triggering leukocyte apoptosis, roscovitine can also enhance the resolution of inflammation. Beyond death-inducing properties, we tested whether roscovitine affects leukocyte-endothelial cell interaction, a vital step in the onset of inflammation. Leukocyte-endothelial cell interactions were evaluated in venules of mouse cremaster muscle, using intravital microscopy. In primary human endothelial cells, we studied the influence of roscovitine on adhesion molecules and on the nuclear factor-κB (NF-κB) pathway. A cellular kinome array, in vitro CDK profiling and RNAi methods were used to identify targets of roscovitine. In vivo, roscovitine attenuated the tumour necrosis factor-α (TNF-α)-induced leukocyte adherence to and transmigration through, the endothelium. In vitro, roscovitine strongly inhibited TNF-α-evoked expression of endothelial adhesion molecules (E-selectin, intercellular cell adhesion molecule, vascular cell adhesion molecule). Roscovitine blocked NF-κB-dependent gene transcription, but not the NF-κB activation cascade [inhibitor of κB (IκB) kinase activity, IκB-α degradation, p65 translocation]. Using a cellular kinome array and an in vitro CDK panel, we found that roscovitine inhibited protein kinase A, ribosomal S6 kinase and CDKs 2, 5, 7 and 9. Experiments using kinase inhibitors and siRNA showed that the decreased endothelial activation was due solely to blockade of CDK5 and CDK9 by roscovitine. Our study highlights a novel mode of action for roscovitine, preventing endothelial activation and leukocyte-endothelial cell interaction by inhibition of CDK5 and 9. This might expand its usage as a promising anti-inflammatory compound. © 2011 The Authors. British Journal of Pharmacology © 2011 The British Pharmacological Society.

  19. Functional role of gap junctions in cytokine-induced leukocyte adhesion to endothelium in vivo

    Science.gov (United States)

    Véliz, Loreto P.; González, Francisco G.; Duling, Brian R.; Sáez, Juan C.; Boric, Mauricio P.

    2008-01-01

    To assess the hypothesis that gap junctions (GJs) participate on leukocyte-endothelium interactions in the inflammatory response, we compared leukocyte adhesion and transmigration elicited by cytokine stimulation in the presence or absence of GJ blockers in the hamster cheek pouch and also in the cremaster muscle of wild-type (WT) and endothelium-specific connexin 43 (Cx43) null mice (Cx43e−/−). In the cheek pouch, topical tumor necrosis factor-α (TNF-α; 150 ng/ml, 15 min) caused a sustained increment in the number of leukocytes adhered to venular endothelium (LAV) and located at perivenular regions (LPV). Superfusion with the GJ blockers 18-α-glycyrrhetinic acid (AGA; 75 μM) or 18-β-glycyrrhetinic acid (50 μM) abolished the TNF-α-induced increase in LAV and LPV; carbenoxolone (75 μM) or oleamide (100 μM) reduced LAV by 50 and 75%, respectively, and LPV to a lesser extent. None of these GJ blockers modified venular diameter, blood flow, or leukocyte rolling. In contrast, glycyrrhizin (75 μM), a non-GJ blocker analog of AGA, was devoid of effect. Interestingly, when AGA was removed 90 min after TNF-α stimulation, LAV started to rise at a similar rate as in control. Conversely, application of AGA 90 min after TNF-α reduced the number of previously adhered cells. In WT mice, intrascrotal injection of TNF-α (0.5 μg/0.3 ml) increased LAV (fourfold) and LPV (threefold) compared with saline-injected controls. In contrast to the observations in WT animals, TNF-α stimulation did not increase LAV or LPV in Cx43e−/− mice. These results demonstrate an important role for GJ communication in leukocyte adhesion and transmigration during acute inflammation in vivo and further suggest that endothelial Cx43 is key in these processes. PMID:18599597

  20. [Dynamic studies of the leukocyte phagocytic activity after exposure of rats to asbestos and basalt fibers].

    Science.gov (United States)

    Hurbánková, M

    1993-05-01

    The paper presents the results of the dynamic one-year follow-up of the phagocytic activity of Wistar-rats peripheral blood leukocytes following intraperitoneal administration of asbestos and basalt fibres (Man-Made Mineral Fibres--MMMF). We investigated the phagocytic activity of leukocytes in peripheral blood following intraperitoneal administration of asbestos and basalt fibres to rats 2, 24, 48 h as well as 1, 2, 4, 8 weeks and 6 and 12 months after dosing. We investigated the time dependent of the changes of relative granulocytes count, percentage of phagocytizing cells from leukocytes, percentage of phagocytizing granulocytes and percentage of phagocytizing monocytes. The results of our experiment showed that asbestos and basalt fibres differed in their effects on the parameters studied. Granulocyte count as well as the phagocytic activity of leukocytes during the one-year dynamic follow-up in both dust--exposed groups of animals were found to change in two phases, characterised by the initial stimulation of the acute phase (I), followed by the suppression of the parameters in the chronic phase (II). Exposure to asbestos and basalt fibres led, in phase II, to impairment of the phagocytic activity of granulocytes. Asbestos fibres at the same time significantly decreased also the phagocytic activity of monocytes. Exposure to basalt fibres did not affect the phagocytic activity of monocytes in phase II. It follows from the results of the experiment, that the monocytic component of leukocytes probably plays an important role in the development of diseases caused by exposure to fibrous dusts and basalt fibres have smaller biological effects compared with asbestos fibres.

  1. Altered mitochondrial function and oxidative stress in leukocytes of anorexia nervosa patients.

    Directory of Open Access Journals (Sweden)

    Victor M Victor

    Full Text Available CONTEXT: Anorexia nervosa is a common illness among adolescents and is characterised by oxidative stress. OBJECTIVE: The effects of anorexia on mitochondrial function and redox state in leukocytes from anorexic subjects were evaluated. DESIGN AND SETTING: A multi-centre, cross-sectional case-control study was performed. PATIENTS: Our study population consisted of 20 anorexic patients and 20 age-matched controls, all of which were Caucasian women. MAIN OUTCOME MEASURES: Anthropometric and metabolic parameters were evaluated in the study population. To assess whether anorexia nervosa affects mitochondrial function and redox state in leukocytes of anorexic patients, we measured mitochondrial oxygen consumption, membrane potential, reactive oxygen species production, glutathione levels, mitochondrial mass, and complex I and III activity in polymorphonuclear cells. RESULTS: Mitochondrial function was impaired in the leukocytes of the anorexic patients. This was evident in a decrease in mitochondrial O2 consumption (P<0.05, mitochondrial membrane potential (P<0.01 and GSH levels (P<0.05, and an increase in ROS production (P<0.05 with respect to control subjects. Furthermore, a reduction of mitochondrial mass was detected in leukocytes of the anorexic patients (P<0.05, while the activity of mitochondrial complex I (P<0.001, but not that of complex III, was found to be inhibited in the same population. CONCLUSIONS: Oxidative stress is produced in the leukocytes of anorexic patients and is closely related to mitochondrial dysfunction. Our results lead us to propose that the oxidative stress that occurs in anorexia takes place at mitochondrial complex I. Future research concerning mitochondrial dysfunction and oxidative stress should aim to determine the physiological mechanism involved in this effect and the physiological impact of anorexia.

  2. Goat uterine DBA+ leukocytes differentiation and cytokines expression respond differently to cloned versus fertilized embryos.

    Directory of Open Access Journals (Sweden)

    Lijuan Qin

    Full Text Available High rate of fetal mortality in ruminant somatic cell nuclear transfer (SCNT pregnancies is due, at least in part, to immune-mediated abortion of fetuses. In the present study, goat uterine leukocytes were isolated by Dolichos biflorus agglutinin (DBA coated magnetic beads, and with majority being were CD56+CD16- in phenotype with low levels of perforin and Granzyme B expression. The responses of the isolated cells to SCNT and in vitro fertilization (IVF embryos conditioned mediums containing hormone steroids were compared by measuring their phenotype and cytokines expression. The results showed there was a 2-fold increase in the numbers of isolated uterine leukocytes after incubation with different conditioned mediums for 120 h. However, significantly lower percentage and absolute numbers of uterine CD56+CD16- leukocytes incubated with SCNT conditioned mediums were detected as compared with those incubated with IVF conditioned mediums (P < 0.05. The group treated with progesterone (P4 or the combination of P4 and 17β-estradiol (E2 were associated with significantly higher percentage and absolute numbers of CD56+CD16- cells as compared with those treated with E2 alone (P < 0.05. Furthermore, in the presence of steroids, the isolated leukocytes incubated with SCNT conditioned mediums associated with greater levels of IFN-γ secretion and expression, as well as lesser levels of VEGF, as compared with those treated with IVF conditioned mediums (P < 0.05. In conclusion, this study demonstrates that SCNT embryos have a profound effect on the phenotype expression of goat uterine DBA+ leukocytes, as well as the secretion and expression of IFN-γ and VEGF by these cells in vitro.

  3. Red blood cell and leukocyte alloimmunization in patients awaiting kidney transplantation

    Science.gov (United States)

    da Silva, Silvia Fernandes Ribeiro; Ferreira, Gláucia Maria; da Silva, Sonia Leite; Alves, Tânia Maria de Oliveira; Ribeiro, Ilana Farias; Ribeiro, Thyciana Rodrigues; Cavalcante, Maria do Carmo Serpa

    2013-01-01

    Objective To determine the rates of red blood cell and leukocyte alloimmunization in patients with chronic kidney disease awaiting kidney transplantation. Methods In this cross-sectional and prospective study, the serum of 393 chronic kidney disease patients on a transplant waiting list in Ceará, Northeastern Brazil were tested for red cell and leukocyte antibodies. In addition, demographic, clinical and laboratory data were collected. Results The average age in the sample of 393 patients was 34.1 ± 14 years. Slightly more than half (208; 52.9%) were male. The average numbers of transfusions and gestations were 3.1 ± 3.3 and 1.6 ± 6, respectively. One third (33.6%) were alloimmunized: 78% with leukocyte antibodies, 9.1% with red cell antibodies and 12.9% with both. Red cell antibodies were detected in 29 cases (7.4%), 17 of whom were women, who had received more transfusions than the males (p-value < 0.0001). The most frequently detected red cell antibodies belonged to the Rh (24.1%) and Kell (13.8%) blood group systems. Leukocyte antibodies were detected in 30.5% of cases, 83 of whom were women, who had received more transfusions than the males (p-value < 0.0001) and were more reactive to panel reactive antibodies (p-value < 0.0001). The mean alloreactivity to panel reactive antibodies was 47.7 ± 31.2%. Conclusion Chronic kidney disease patients on the transplant waiting list in Ceará, Brazil, display high rates of red cell (7.4%) and leukocyte (30.5%) alloimmunization. In this sample, alloimmunization was significantly associated with the number of transfusions and gender. PMID:23904808

  4. Association of postmenopausal endogenous sex hormones with global methylation level of leukocyte DNA among Japanese women

    Directory of Open Access Journals (Sweden)

    Iwasaki Motoki

    2012-07-01

    Full Text Available Abstract Background Although global hypomethylation of leukocyte DNA has been associated with an increased risk of several sites of cancer, including breast cancer, determinants of global methylation level among healthy individuals remain largely unexplored. Here, we examined whether postmenopausal endogenous sex hormones were associated with the global methylation level of leukocyte DNA. Methods A cross-sectional study was conducted using the control group of a breast cancer case–control study in Nagano, Japan. Subjects were postmenopausal women aged 55 years or over who provided blood samples. We measured global methylation level of peripheral blood leukocyte DNA by luminometric methylation assay; estradiol, estrone, androstenedione, dehydroepiandrosterone sulfate, testosterone and free testosterone by radioimmunoassay; bioavailable estradiol by the ammonium sulfate precipitation method; and sex-hormone binding globulin by immunoradiometric assay. A linear trend of association between methylation and hormone levels was evaluated by regression coefficients in a multivariable liner regression model. A total of 185 women were included in the analyses. Results Mean global methylation level (standard deviation was 70.3% (3.1 and range was from 60.3% to 79.2%. Global methylation level decreased 0.27% per quartile category for estradiol and 0.39% per quartile category for estrone while it increased 0.41% per quartile category for bioavailable estradiol. However, we found no statistically significant association of any sex hormone level measured in the present study with global methylation level of leukocyte DNA. Conclusions Our findings suggest that endogenous sex hormones are not major determinants of the global methylation level of leukocyte DNA.

  5. Determination of an unrelated donor pool size for human leukocyte antigen-matched platelets in Brazil

    Directory of Open Access Journals (Sweden)

    Carolina Bonet Bub

    2016-02-01

    Full Text Available ABSTRACT Background: Successful transfusion of platelet refractory patients is a challenge. Many potential donors are needed to sustain human leukocyte antigen matched-platelet transfusion programs because of the different types of antigens and the constant needs of these patients. For a highly mixed population such as the Brazilian population, the pool size required to provide adequate platelet support is unknown. Methods: A mathematical model was created to estimate the appropriate size of an unrelated donor pool to provide human leukocyte antigen-compatible platelet support for a Brazilian population. A group of 154 hematologic human leukocyte antigen-typed patients was used as the potential patient population and a database of 65,500 human leukocyte antigen-typed bone marrow registered donors was used as the donor population. Platelet compatibility was based on the grading system of Duquesnoy. Results: Using the mathematical model, a pool containing 31,940, 1710 and 321 donors would be necessary to match more than 80% of the patients with at least five completely compatible (no cross-reactive group, partial compatible (one cross-reactive group or less compatible (two cross-reactive group donors, respectively. Conclusion: The phenotypic diversity of the Brazilian population has probably made it more difficulty to find completely compatible donors. However, this heterogeneity seems to have facilitated finding donors when cross-reactive groups are accepted as proposed by the grading system of Duquesnoy. The results of this study may help to establish unrelated human leukocyte antigen-compatible platelet transfusions, a procedure not routinely performed in most Brazilian transfusion services.

  6. Increased expression of telomere-regulating genes in endurance athletes with long leukocyte telomeres.

    Science.gov (United States)

    Denham, Joshua; O'Brien, Brendan J; Prestes, Priscilla R; Brown, Nicholas J; Charchar, Fadi J

    2016-01-15

    Leukocyte telomeres shorten with age, and excessive shortening is associated with age-related cardiometabolic diseases. Exercise training may prevent disease through telomere length maintenance although the optimal amount of exercise that attenuates telomere attrition is unknown. Furthermore, the underlying molecular mechanisms responsible for the enhanced telomere maintenance observed in endurance athletes is poorly understood. We quantified the leukocyte telomere length and analyzed the expression of telomere-regulating genes in endurance athletes and healthy controls (both n = 61), using quantitative PCR. We found endurance athletes have significantly longer (7.1%, 208-416 nt) leukocyte telomeres and upregulated TERT (2.0-fold) and TPP1 (1.3-fold) mRNA expression compared with controls in age-adjusted analysis. The telomere length and telomere-regulating gene expression differences were no longer statistically significant after adjustment for resting heart rate and relative V̇O(2 max) (all P > 0.05). Resting heart rate emerged as an independent predictor of leukocyte telomere length and TERT and TPP1 mRNA expression in stepwise regression models. To gauge whether volume of exercise was associated with leukocyte telomere length, we divided subjects into running and cycling tertiles (distance covered per week) and found individuals in the middle and highest tertiles had longer telomeres than individuals in the lowest tertile. These data emphasize the importance of cardiorespiratory fitness and exercise training in the prevention of biological aging. They also support the concept that moderate amounts of exercise training protects against biological aging, while higher amounts may not elicit additional benefits.

  7. Leukocyte telomere length in major depression: correlations with chronicity, inflammation and oxidative stress--preliminary findings.

    Directory of Open Access Journals (Sweden)

    Owen M Wolkowitz

    Full Text Available BACKGROUND: Depression is associated with an unusually high rate of aging-related illnesses and early mortality. One aspect of "accelerated aging" in depression may be shortened leukocyte telomeres. When telomeres critically shorten, as often occurs with repeated mitoses or in response to oxidation and inflammation, cells may die. Indeed, leukocyte telomere shortening predicts early mortality and medical illnesses in non-depressed populations. We sought to determine if leukocyte telomeres are shortened in Major Depressive Disorder (MDD, whether this is a function of lifetime depression exposure and whether this is related to putative mediators, oxidation and inflammation. METHODOLOGY: Leukocyte telomere length was compared between 18 unmedicated MDD subjects and 17 controls and was correlated with lifetime depression chronicity and peripheral markers of oxidation (F2-isoprostane/Vitamin C ratio and inflammation (IL-6. Analyses were controlled for age and sex. PRINCIPAL FINDINGS: The depressed group, as a whole, did not differ from the controls in telomere length. However, telomere length was significantly inversely correlated with lifetime depression exposure, even after controlling for age (p<0.05. Average telomere length in the depressed subjects who were above the median of lifetime depression exposure (≥9.2 years' cumulative duration was 281 base pairs shorter than that in controls (p<0.05, corresponding to approximately seven years of "accelerated cell aging." Telomere length was inversely correlated with oxidative stress in the depressed subjects (p<0.01 and in the controls (p<0.05 and with inflammation in the depressed subjects (p<0.05. CONCLUSIONS: These preliminary data indicate that accelerated aging at the level of leukocyte telomeres is proportional to lifetime exposure to MDD. This might be related to cumulative exposure to oxidative stress and inflammation in MDD. This suggest that telomere shortening does not antedate depression

  8. Healthy lifestyle and leukocyte telomere length in U.S. women.

    Directory of Open Access Journals (Sweden)

    Qi Sun

    Full Text Available CONTEXT: Whether a healthy lifestyle may be associated with longer telomere length is largely unknown. OBJECTIVES: To examine healthy lifestyle practices, which are primary prevention measures against major age-related chronic diseases, in relation to leukocyte telomere length. DESIGN AND SETTING: Cross-sectional analysis in the Nurses' Health Study (NHS. PARTICIPANTS: The population consisted of 5,862 women who participated in multiple prospective case-control studies within the NHS cohort. Z scores of leukocyte telomere length were derived within each case-control study. Based on prior work, we defined low-risk or healthy categories for five major modifiable factors assessed in 1988 or 1990: non-current smoking, maintaining a healthy body weight (body mass index in 18.5-24.9 kg/m(2, engaging in regular moderate or vigorous physical activities (≥150 minutes/week, drinking alcohol in moderation (1 drink/week to <2 drinks/day, and eating a healthy diet (Alternate Healthy Eating Index score in top 50%. We calculated difference (% of the z scores contrasting low-risk groups with reference groups to evaluate the association of interest. RESULTS: Although none of the individual low-risk factors was significantly associated with larger leukocyte telomere length z scores, we observed a significant, positive relationship between the number of low-risk factors and the z scores. In comparison with women who had zero low-risk factors (1.9% of the total population and were, therefore, considered the least healthy group, the leukocyte telomere length z scores were 16.4%, 22.1%, 28.7%, 22.6%, and 31.2% (P for trend = 0.015 higher for women who had 1 to 5 low-risk factors, respectively. CONCLUSIONS: Adherence to a healthy lifestyle, defined by major modifiable risk factors, was associated with longer telomere length in leukocytes.

  9. SRC protein tyrosine kinase, c-Jun N-terminal kinase (JNK), and NF-kappaBp65 signaling in commercial and wild-type turkey leukocytes

    Science.gov (United States)

    Studies comparing signaling in wild-type turkey (WT) leukocytes and commercial turkey (CT) leukocytes found that the activity of protein tyrosine kinases (PTK) and MAP kinases, ERK 1/2 and p38, were significantly higher in WT leukocytes compared to CT lines upon exposure to both SE and OPSE on days...

  10. Relative Leukocyte Telomere Length, Hematological Parameters and Anemia - Data from the Berlin Aging Study II (BASE-II).

    Science.gov (United States)

    Meyer, Antje; Salewsky, Bastian; Buchmann, Nikolaus; Steinhagen-Thiessen, Elisabeth; Demuth, Ilja

    2016-01-01

    The length of the chromosome ends, telomeres, is widely accepted as a biomarker of aging. However, the dynamic of the relationship between telomere length and hematopoietic parameters in the normal aging process, which is of particular interest with respect to age-related anemia, is not well understood. We have analyzed the relationship between relative leukocyte telomere length (rLTL) and several hematological parameters in the older group of the Berlin Aging Study II (BASE-II) participants. This paper also compares rLTL between both BASE-II age groups (22-37 and 60-83 years). Genomic DNA was extracted from peripheral blood leukocytes of BASE-II participants and used to determine rLTL by a quantitative PCR protocol. Standard methods were used to determine blood parameters, and the WHO criteria were used to identify anemic participants. Telomere length data were available for 444 younger participants (28.4 ± 3.1 years old; 52% women) and 1,460 older participants (68.2 ± 3.7 years old; 49.4% women). rLTL was significantly shorter in BASE-II participants of the older group (p = 3.7 × 10-12) and in women (p = 4.2 × 10-31). rLTL of older men exhibited a statistically significant, positive partial correlation with mean corpuscular hemoglobin (MCH; p = 0.012) and MCH concentration (p = 0.002). While these correlations were only observed in men, the rLTL of older women was negatively correlated with the number of thrombocytes (p = 0.015) in the same type of analysis. Among all older participants, 6% met the criteria to be categorized as 'anemic'; however, there was no association between anemia and rLTL. In the present study, we have detected isolated correlations between rLTL and hematological parameters; however, in all cases, rLTL explained only a small part of the variation of the analyzed parameters. In disagreement with some other studies showing similar data, we interpret the association between rLTL and some of the hematological parameters studied here to be

  11. [Effect of herba schizonepetae volatile oil (STO) on activity of 5-lipoxygenase in rat thoracic cavity leukocytes].

    Science.gov (United States)

    Zhao, Lu; Zeng, Nan; Tang, Yong-xin; Liu, Xiao-shuai; Yang, Xuan; Qu, Li-ping

    2008-09-01

    To investigate the effect of herba schizonepetae volatile oil (STO) on the activity of 5-lipoxygenase (5-LO), so as to elucidate its mechanisms of anti-inflammatory action which is related to the arachidonic acid (AA) metabolism. Thoracic cavity leukocytes from the pleurisy model rat induced by injecting 1%-carrageenan into the pleural cavity were collected. Then 0. 4 mL cell suspension including 2 x 10(7) cells per millilitre were used as the reaction system in vitro. STO in different concentrations (final concentration 0.011, 0.022, 0.043, 0.087, 0.179, 0.255, 0.364 g x L(-1)), zileuton (final concentration 0.625 x 10(-3) g x L(-1)), and DMSO in the same volume were added into the reaction tube respectively. The reaction tubes were incubated at 37 degrees C for 20 min and CaCl2 (final concentration 2 mmol x L(-1)), MgCl2 (final concentration 0.5 mmol x L(-1)), exogenous AA (final concentration 200 micromol x L(-1)) and A23187 (final concentration 5 micromol x L(-1)) were added in turns during this period. The reaction tubes were mixed and continuously incubated at 37 degrees C for 30 min. After terminating reaction by adding methanol, the metabolites of 5-LO, leukotriene B4 (LTB4) and 5-hydroxy-6, 8, 11, 14-eicosatetraenoic acid (5-HETE), were extracted, separated and detected by means of RP-HPLC. Compared with control group, STO significantly inhibited the biosynthesis of LTB4 and 5-HETE at final concentration between 0. 022 g x L(-1) and 0.364 g x L(-1) (P STO can inhibited the activity of 5-LO, which is an important enzyme of AA metabolism, in rat thoracic cavity leukocytes in a dose-dependent manner in vitro. It is suggested that the mechanism of anti-inflammatory action of STO is related to its inhibiting the activity of 5-LO and decreasing the level of major inflammatory mediators LTB4.

  12. The role of leukocytes from L-PRP/L-PRF in wound healing and immune defense: new perspectives.

    Science.gov (United States)

    Bielecki, Tomasz; Dohan Ehrenfest, David M; Everts, Peter A; Wiczkowski, Andrzej

    2012-06-01

    Platelet concentrates for topical use are innovative tools of regenerative medicine and their effects in various therapeutical situations are hotly debated. Unfortunately, this field of research mainly focused on the platelet growth factors, and the fibrin architecture and the leukocyte content of these products are too often neglected. In the four families of platelet concentrates, 2 families contain significant concentrations of leukocytes: L-PRP (Leukocyte- and Platelet-Rich Plasma) and L-PRF (Leukocyte- and Platelet-Rich Fibrin). The presence of leukocytes has a great impact on the biology of these products, not only because of their immune and antibacterial properties, but also because they are turntables of the wound healing process and the local factor regulation. In this article, the various kinds of leukocytes present in a platelet concentrate are described (particularly the various populations of granulocytes and lymphocytes), and we insist on the large diversity of factors and pathways that these cells can use to defend the wound site against infections and to regulate the healing process. Finally, the impact of these cells in the healing properties of the L-PRP and L-PRF is also discussed: if antimicrobial properties were already pointed out, effects in the regulation of cell proliferation and differentiation were also hypothesized. Leukocytes are key actors of many platelet concentrates, and a better understanding of their effects is an important issue for the development of these technologies.

  13. RAGE and ICAM-1 differentially control leukocyte recruitment during acute inflammation in a stimulus-dependent manner

    Directory of Open Access Journals (Sweden)

    Nawroth Peter P

    2011-10-01

    Full Text Available Abstract Background The receptor for advanced glycation endproducts, RAGE, is involved in the pathogenesis of many inflammatory conditions, which is mostly related to its strong activation of NF-κB but also due to its function as ligand for the β2-integrin Mac-1. To further dissect the stimulus-dependent role of RAGE on leukocyte recruitment during inflammation, we investigated β2-integrin-dependent leukocyte adhesion in RAGE-/- and Icam1-/- mice in different cremaster muscle models of inflammation using intravital microscopy. Results We demonstrate that RAGE, but not ICAM-1 substantially contributes to N-formyl-methionyl-leucyl-phenylalanine (fMLP-induced leukocyte adhesion in TNF-α-pretreated cremaster muscle venules in a Mac-1-dependent manner. In contrast, fMLP-stimulated leukocyte adhesion in unstimulated cremaster muscle venules is independent of RAGE, but dependent on ICAM-1 and its interaction with LFA-1. Furthermore, chemokine CXCL1-stimulated leukocyte adhesion in surgically prepared cremaster muscle venules was independent of RAGE but strongly dependent on ICAM-1 and LFA-1 suggesting a differential and stimulus-dependent regulation of leukocyte adhesion during inflammation in vivo. Conclusion Our results demonstrate that RAGE and ICAM-1 differentially regulate leukocyte adhesion in vivo in a stimulus-dependent manner.

  14. Involvement of seminal leukocytes, reactive oxygen species, and sperm mitochondrial membrane potential in the DNA damage of the human spermatozoa.

    Science.gov (United States)

    Lobascio, A M; De Felici, M; Anibaldi, M; Greco, P; Minasi, M G; Greco, E

    2015-03-01

    Measurement of reactive oxygen species (ROS) producing leukocytes in semen has been a standard component of the semen analysis, but its true significance remains still unknown. In this study, we have correlated the number of seminal leukocytes to various semen parameters. We found a negative correlation between the leukocyte number and sperm concentration (rs  = -0.22; p = 0.01) and motility (rs  = -0.20; p = 0.02). In contrast, a positive correlation between the number of leukocytes and both seminal ROS (rs  = 0.70, p sperm mitochondrial membrane potential (MMP) (10% vs 35%, rs  = 0.25, p = 0.08; n = 50). Overall these results indicate that the presence of high number of leukocytes in the ejaculate negatively affects key semen parameters, as sperm concentration and motility, associated with infertility conditions. Moreover, they suggest that leukocytes are the major source of the seminal ROS and cause of sperm DNA fragmentation. However, the absence of a clear correlation between ROS and sperm DNA fragmentation, and spermatozoa with damaged DNA and MMP loss, suggest that ROS produced by leukocytes might be not the only cause of DNA damage in spermatozoa and that intrinsic mitochondrial-dependent apoptotic pathways might not have a major impact on sperm DNA fragmentation.

  15. Anethole and eugenol reduce in vitro and in vivo leukocyte migration induced by fMLP, LTB4, and carrageenan.

    Science.gov (United States)

    Estevão-Silva, Camila Fernanda; Kummer, Raquel; Fachini-Queiroz, Fernanda Carolina; Grespan, Renata; Nogueira de Melo, Gessilda Alcântara; Baroni, Silmara; Cuman, Roberto Kenji Nakamura; Bersani-Amado, Ciomar Aparecida

    2014-07-01

    The aim of this study was to investigate the effect of anethole (AN) and eugenol (EUG) on leukocyte migration using in vitro chemotaxis and in situ microcirculation assays. BALB/c mice were used for the in vitro chemotaxis assay, and Wistar rats for the in situ microcirculation assay. We evaluated (a) the in vitro leukocyte migration in response to chemotactic factors (formyl-methionyl-leucyl-phenylalanine [fMLP] and leukotriene B4 [LTB4]) and (b) the rolling, adhesion, and migration of leukocytes induced by an injection of carrageenan (100 µg/cavity) into the scrotum of the animal. In the in vitro chemotaxis assay, AN and EUG at doses of 1, 3, 9, and 27 µg/ml significantly inhibited leukocyte migration when stimulated by the chemotactic agents fMLP and LTB4. In the in situ microcirculation assay, AN at doses of 125 and 250 mg/kg and EUG at a dose of 250 mg/kg significantly decreased the number of leukocytes that rolled, adhered, and migrated to perivascular tissue. The results indicate that AN and EUG exert inhibitory effects on leukocyte migration, highlighting their possible use to diminish excessive leukocyte migration in the inflammatory process.

  16. Calibrating spatio-temporal models of leukocyte dynamics against in vivo live-imaging data using approximate Bayesian computation

    Science.gov (United States)

    Barnes, Chris P.; Huvet, Maxime; Bugeon, Laurence; Thorne, Thomas; Lamb, Jonathan R.; Dallman, Margaret J.; Stumpf, Michael P. H.

    2016-01-01

    In vivo studies allow us to investigate biological processes at the level of the organism. But not all aspects of in vivo systems are amenable to direct experimental measurements. In order to make the most of such data we therefore require statistical tools that allow us to obtain reliable estimates for e.g. kinetic in vivo parameters. Here we show how we can use approximate Bayesian computation approaches in order to analyse leukocyte migration in zebrafish embryos in response to injuries. We track individual leukocytes using live imaging following surgical injury to the embryos’ tail-fins. The signalling gradient that leukocytes follow towards the site of the injury cannot be directly measured but we can estimate its shape and how it changes with time from the directly observed patterns of leukocyte migration. By coupling simple models of immune signalling and leukocyte migration with the unknown gradient shape into a single statistical framework we can gain detailed insights into the tissue-wide processes that are involved in the innate immune response to wound injury. In particular we find conclusive evidence for a temporally and spatially changing signalling gradient that modulates the changing activity of the leukocyte population in the embryos. We conclude with a robustness analysis which highlights the most important factors determining the leukocyte dynamics. Our approach relies only on the ability to simulate numerically the process under investigation and is therefore also applicable in other in vivo contexts and studies. PMID:22327539

  17. Protein Extractability

    African Journals Online (AJOL)

    human nutrition, particularly in developing ... studied the effect of NaCl on the extractability of ... significant influence on the functional properties ..... Malaka, S. L. O. and Amund, O. O. Studies on the Life Cycle and Morphometrics of Honeybees,.

  18. Leukocyte counts in urine reflect the risk of concomitant sepsis in bacteriuric infants: A retrospective cohort study

    Directory of Open Access Journals (Sweden)

    Harper Marvin B

    2007-06-01

    Full Text Available Abstract Background When urine infections are missed in febrile young infants with normal urinalysis, clinicians may worry about the risk – hitherto unverified – of concomitant invasion of blood and cerebrospinal fluid by uropathogens. In this study, we determine the extent of this risk. Methods In a retrospective cohort study of febrile 0–89 day old infants evaluated for sepsis in an urban academic pediatric emergency department (1993–1999, we estimated rates of bacteriuric sepsis (urinary tract infections complicated by sepsis after stratifying infants by urine leukocyte counts higher, or lower than 10 cells/hpf. We compared the global accuracy of leukocytes in urine, leukocytes in peripheral blood, body temperature, and age for predicting bacteruric sepsis. The global accuracy of each test was estimated by calculating the area under its receiver operating characteristic curve (AUC. Chi-square and Fisher exact tests compared count data. Medians for data not normally distributed were compared by the Kruskal-Wallis test. Results Two thousand two hundred forty-nine young infants had a normal screening dipstick. None of these developed bacteremia or meningitis despite positive urine culture in 41 (1.8%. Of 1516 additional urine specimens sent for formal urinalysis, 1279 had 0–9 leukocytes/hpf. Urine pathogens were isolated less commonly (6% vs. 76% and at lower concentrations in infants with few, compared to many urine leukocytes. Urine leukocytes (AUC: 0.94 were the most accurate predictors of bacteruric sepsis. Infants with urinary leukocytes Conclusion In young infants evaluated for fever, leukocytes in urine reflect the likelihood of bacteriuric sepsis. Infants with urinary tract infections missed because of few leukocytes in urine are at relatively low risk of invasive bacterial sepsis by pathogens isolated from urine.

  19. Flow cytofluorometric assay of human whole blood leukocyte DNA degradation in response to Yersinia pestis and Staphylococcus aureus

    Science.gov (United States)

    Kravtsov, Alexander L.; Grebenyukova, Tatyana P.; Bobyleva, Elena V.; Golovko, Elena M.; Malyukova, Tatyana A.; Lyapin, Mikhail N.; Kostyukova, Tatyana A.; Yezhov, Igor N.; Kuznetsov, Oleg S.

    2001-05-01

    Human leukocytes containing less than 2C DNA per cell (damaged or dead cells) were detected and quantified by flow cytometry and DNA-specific staining with ethidium bromide and mithramycin in whole blood infected with Staphylococcus aureus or Yersinia pestis. Addition of live S. aureus to the blood (100 microbe cells per one leukocyte) resulted in rapid degradation of leukocyte DNA within 3 to 6 hours of incubation at 37 degree(s)C. However, only about 50 percent cells were damaged and the leukocytes with the intact genetic apparatus could be found in the blood for a period up to 24 hours. The leukocyte injury was preceded by an increase of DNA per cell content (as compared to the normal one) that was likely to be connected with the active phagocytosis of S. aureus by granulocytes (2C DNA of diploid phagocytes plus the all bacterial DNA absorbed). In response to the same dose of actively growing (at 37 degree(s)C) virulent Y. pestis cells, no increase in DNA content per cell could be observed in the human blood leukocytes. The process of the leukocyte DNA degradation started after a 6-hour incubation, and between 18 to 24 hours of incubation about 90 percent leukocytes (phagocytes and lymphocytes) lost their specific DNA fluorescence. These results demonstrated a high potential of flow cytometry in comparative analysis in vitro of the leukocyte DNA degradation process in human blood in response to bacteria with various pathogenic properties. They agree with the modern idea of an apoptotic mechanism of immunosuppression in plague.

  20. Bevalac extraction

    Energy Technology Data Exchange (ETDEWEB)

    Kalnins, J.G.; Krebs, G.; Tekawa, M.; Cowles, D.; Byrne, T.

    1992-02-01

    This report will describe some of the general features of the Bevatron extraction system, primarily the dependence of the beam parameters and extraction magnet currents on the Bevalac field. The extraction magnets considered are: PFW, XPl, XP2, XS1, XS2, XM1, XM2, XM3, XQ3A and X03B. This study is based on 84 past tunes (from 1987 to the present) of various ions (p,He,O,Ne,Si,S,Ar,Ca,Ti,Fe,Nb,La,Au and U), for Bevalac fields from 1.749 to 12.575 kG, where all tunes included a complete set of beam line wire chamber pictures. The circulating beam intensity inside the Bevalac is measured with Beam Induction Electrodes (BIE) in the South Tangent Tank. The extracted beam intensity is usually measured with the Secondary Emission Monitor (SEM) in the F1-Box. For most of the tunes the extraction efficiency, as given by the SEM/BIE ratio, was not recorded in the MCR Log Book, but plotting the available Log Book data as a function of the Bevalac field, see Fig.9, we find that the extraction efficiency is typically between 30->60% with feedback spill.

  1. Apoptosis of peripheral blood leukocytes from rabbits infected with non-haemagglutinating strains of rabbit haemorrhagic disease virus (RHDV).

    Science.gov (United States)

    Niedźwiedzka-Rystwej, Paulina; Deptuła, Wiesław

    2012-09-15

    The report demonstrates that the induction of apoptosis in peripheral blood granulocytes and lymphocytes of rabbits infected with three non-haemagglutinating RHDV strains (English Rainham, German Frankfurt, and Spanish Asturias) is a crucial determinant of the pathogenesis of rabbit haemorrhagic disease. Apoptosis was measured by flow cytometric detection of caspase activity. These studies demonstrated that the investigated RHDV (rabbit haemorrhagic disease virus) viral strains affected leukocyte apoptosis to varying degrees. Enhanced leukocyte apoptosis was detected between 4 and 36 h after infection and was more pronounced in lymphocytes than in granulocytes. The data presented here thus provide a preliminary understanding of the kinetics of apoptosis in leukocytes of rabbits infected with RHDV.

  2. Primary characterization and evaluation of anti ulcerogenic activity of an aqueous extract from callus culture of Cereus peruvianus Mill. (Cactaceae).

    Science.gov (United States)

    Jayme, Milena O; Ames, Franciele Q; Bersani-Amado, Ciomar A; Machado, Maria de Fatima P S; Mangolin, Claudete A; Goncalves, Regina A C; de Oliveira, Arildo J B

    2015-01-01

    In the current study we reported cultivation, extraction procedure, analysis and preliminary characterization of the aqueous extract from Cereus peruvianus callus culture and evaluated its anti ulcerogenic activity in vivo models of experimental ulcers in Wistar rats. The obtained aqueous extract from callus (AC) was dialyzed and subjected to freeze-thaw process, providing a possible polysaccharide. The carbohydrate and protein contents of the aqueous extract were estimated at 53.4% and 0.66%, respectively, composed primarily of galactose, arabinose and galacturonic acid, with minor amounts of glucose. This appeared heterogeneous when analyzed by high-performance size-exclusion chromatography and a multiangle laser light scattering detector (HPSEC-MALLS). The AC was found to be significantly effective against ethanol-induced lesions but was ineffective against indomethacin-induced lesions. The callus culture of C. peruvianus is an alternative source for the synthesis of substances originally produced by plants. The calluses grown indefinitely in vitro under controlled conditions are stable tissues, and the aqueous extract from calluses may be used instead of fully developed plants using the protocols described in this study.

  3. Influence of Helleborus odorus W. et K. underground parts extracts on red blood pictures of Wistar rats

    Directory of Open Access Journals (Sweden)

    Davidović Vesna

    2010-01-01

    Full Text Available This paper presents the results of investigations of parameter values of the red blood cell count in Wistar rats, after intramuscular application of rhizome and root extract of Helleborus odorus W. et K (HE. The trial was conducted in experimental conditions on 49 Wistar rats divided into 7 groups with 7 animals per unit (control and six experimental groups. The control group was treated by an intramuscular injection of sterile saline (0.25 ml/100 g BM. For the purpose of determing the duration of the HE effects, the animals were injected with a dose of 10 mg/100 g BMand blood was collected by cardial taps after 24, 48 and 72 hrs. Moreover, we investigated the influence of elevated temperatures on the noted HE extract effects, 24 hrs after the application of the inactivated (56oC 30 min or denaturized (100oC 5 min extract, as well as the influence of dialyzed HE. The obtained results of haematological analyses have pointed out the weak hemolyses activity of the extract. In all experimental groups, the erythrocyte count was decreased, but remained within reference values. Haemoglobin concentration and haematocrit values in the experimental and the control groups of rats were decreased in comparison to the physiological values, while the applied extract did not significantly affect the examined parameters. .

  4. Extraction Optimization, Preliminary Characterization and Bioactivities in Vitro of Ligularia hodgsonii Polysaccharides

    Directory of Open Access Journals (Sweden)

    Xueping Song

    2016-05-01

    Full Text Available The optimization extraction, preliminary characterization and bioactivities of Ligularia hodgsonii polysaccharides were investigated. Based on single-factor experiments and orthogonal array test, the optimum extraction conditions were obtained as follows: extraction time 3 h, temperature 85 °C, water/raw material ratio 36. Further Sevag deproteinization and dialysis yielded the dialyzed Ligularia hodgsonii polysaccharides (DLHP, 19.2 ± 1.4 mg/g crude herb. Compositional analysis, size-exclusion chromatography connected with multi-angle laser light-scattering and refractive index (SEC-MALLS-RI, Fourier transform infrared (FT-IR and 1H nuclear magnetic resonance (NMR spectroscopy were employed for characterization of the polysaccharides. DLHP was found to have a major component with a weight-average molecular weight of 1.17 × 105 Da, mainly comprising of glucose, galactose, arabinose, mannose, rhamnose, glucuronic acid and galacturonic acid. By in vitro antioxidant activity assays, DLHP presented remarkable scavenging capacities towards 1,1-diphenyl-2-picrylhydrazyl (DPPH, 2,2′-azinobis (3-ethylbenzothiazoline-6-sulfonic acid (ABTS and hydroxyl radicals, and ferrous ions chelating ability. Moreover, it exhibited appreciable anti-hyperglycemic activity as demonstrated by differential inhibition of α-glucosidase and α-amylase. The results indicated that DLHP could potentially be a resource for antioxidant and hypoglycemic agents.

  5. Purification and properties of Cu-Zn superoxide dismutase extracted from Brucella abortus strain 19

    Energy Technology Data Exchange (ETDEWEB)

    Tabatabai, L.B. (ARS-USDA, Ames, IA (United States))

    1991-03-11

    Recent work showed that a recombinant 20 kDa protein from Brucella abortus expressed in E. coli is a Cu-Zn superoxide dismutase (SOD). Western blot and ELISA results indicated that cattle with brucellosis have antibody to SOD. Here the authors report the purification and properties of the native B. abortus Cu-Zn SOD. SOD was extracted from methanol-killed Brucella abortus strain 19 with 0.1 M sodium citrate-1.0 M sodium chloride solution. The extract was dialyzed and protein precipitated by ammonium sulfate at 70-100% saturation was collected. The SOD was purified by HPLC anion exchange chromatography. SOD activity was assayed with a coupled enzyme assay using xanthine oxidase-cytochrome C reduction assay. The authors determined that the Brucella SOD is present in two molecular forms both inhibitable with KCN with Ki's of 0.32 mM and 4.98 mM, respectively. No other form of SOD was identified in the extract. Polyclonal antibody to SOD and polyclonal antibody to SOD synthetic peptide residues 134-143 inhibited SOD activity by 50% and 13%, respectively. Both SOD and the synthetic peptide inhibited binding of anti-SOD antibody to SOD by 60% and 20%, respectively. Based on these results the SOD and its amphipathic peptide will be considered as candidates for the design of synthetic multiple peptide vaccines and diagnostic reagents for bovine brucellosis.

  6. Purification and Characterization of Fibrinolytic Proteases from Mushroom Volvariela volvaceae Extract.

    Directory of Open Access Journals (Sweden)

    Willy Praira

    2010-11-01

    Full Text Available Purification and Characterization of Fibrinolytic Proteases from Mushroom Volvariela volvaceae Extract. Ediblestraw mushroom (V. volvaceae has been known used for improvement of blood circulation due to its fibrinolyticcontent. The objective of the study is to purify and characterize fibrinolytic protease from straw mushroom extract.Purification were performed through several steps, i.e. precipitation using ammonium sulphate 75%, dialyzed membran(cut-off 10 kDa, and ion-exchange chromatography using DEAE Sepharose. The active fraction of DEAE-Sepharosecontains two purified protein bands with molecular weight of 12.9 and 15.8 kDa. The active fraction has specificactivity of 0.383 U/mg with 2.7 fold higher purification compared to its crude extract. Both crude and purified enzymeshad optimum activity at temperature of 50 ºC and pH 7 in 10 minutes of incubation. Fibrin zymographic profiledemonstrated that the enzyme hydrolyzed fibrin, as well as casein, indicating their potent fibrinolytic activity. Theenzyme was strongly inhibited by phenilmethylsulphonyl fluoride and N-p-tosil-L-lysinchloromethyl keton. Thissuggested that it was a serine protease. In summary, these results showed that crude and purified protease of strawmushroom (V. volvaceae has fibrinolytic activities that can be applied for alternative thrombolytic therapy.

  7. Extraction Optimization, Preliminary Characterization and Bioactivities in Vitro of Ligularia hodgsonii Polysaccharides.

    Science.gov (United States)

    Song, Xueping; Tang, Jun

    2016-05-21

    The optimization extraction, preliminary characterization and bioactivities of Ligularia hodgsonii polysaccharides were investigated. Based on single-factor experiments and orthogonal array test, the optimum extraction conditions were obtained as follows: extraction time 3 h, temperature 85 °C, water/raw material ratio 36. Further Sevag deproteinization and dialysis yielded the dialyzed Ligularia hodgsonii polysaccharides (DLHP, 19.2 ± 1.4 mg/g crude herb). Compositional analysis, size-exclusion chromatography connected with multi-angle laser light-scattering and refractive index (SEC-MALLS-RI), Fourier transform infrared (FT-IR) and ¹H nuclear magnetic resonance (NMR) spectroscopy were employed for characterization of the polysaccharides. DLHP was found to have a major component with a weight-average molecular weight of 1.17 × 10⁵ Da, mainly comprising of glucose, galactose, arabinose, mannose, rhamnose, glucuronic acid and galacturonic acid. By in vitro antioxidant activity assays, DLHP presented remarkable scavenging capacities towards 1,1-diphenyl-2-picrylhydrazyl (DPPH), 2,2'-azinobis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) and hydroxyl radicals, and ferrous ions chelating ability. Moreover, it exhibited appreciable anti-hyperglycemic activity as demonstrated by differential inhibition of α-glucosidase and α-amylase. The results indicated that DLHP could potentially be a resource for antioxidant and hypoglycemic agents.

  8. Comparative genome analysis of three eukaryotic parasites with differing abilities to transform leukocytes reveals key mediators of theileria-induced leukocyte transformation

    KAUST Repository

    Hayashida, Kyoko

    2012-09-04

    We sequenced the genome of Theileria orientalis, a tick-borne apicomplexan protozoan parasite of cattle. The focus of this study was a comparative genome analysis of T. orientalis relative to other highly pathogenic Theileria species, T. parva and T. annulata. T. parva and T. annulata induce transformation of infected cells of lymphocyte or macrophage/monocyte lineages; in contrast, T. orientalis does not induce uncontrolled proliferation of infected leukocytes and multiplies predominantly within infected erythrocytes. While synteny across homologous chromosomes of the three Theileria species was found to be well conserved overall, subtelomeric structures were found to differ substantially, as T. orientalis lacks the large tandemly arrayed subtelomere-encoded variable secreted protein-encoding gene family. Moreover, expansion of particular gene families by gene duplication was found in the genomes of the two transforming Theileria species, most notably, the TashAT/TpHN and Tar/Tpr gene families. Gene families that are present only in T. parva and T. annulata and not in T. orientalis, Babesia bovis, or Plasmo-dium were also identified. Identification of differences between the genome sequences of Theileria species with different abilities to transform and immortalize bovine leukocytes will provide insight into proteins and mechanisms that have evolved to induce and regulate this process. The T. orientalis genome database is available at http://totdb.czc.hokudai.ac.jp/. 2012 Hayashida et al. T.

  9. Comparative genome analysis of three eukaryotic parasites with differing abilities to transform leukocytes reveals key mediators of Theileria-induced leukocyte transformation.

    Science.gov (United States)

    Hayashida, Kyoko; Hara, Yuichiro; Abe, Takashi; Yamasaki, Chisato; Toyoda, Atsushi; Kosuge, Takehide; Suzuki, Yutaka; Sato, Yoshiharu; Kawashima, Shuichi; Katayama, Toshiaki; Wakaguri, Hiroyuki; Inoue, Noboru; Homma, Keiichi; Tada-Umezaki, Masahito; Yagi, Yukio; Fujii, Yasuyuki; Habara, Takuya; Kanehisa, Minoru; Watanabe, Hidemi; Ito, Kimihito; Gojobori, Takashi; Sugawara, Hideaki; Imanishi, Tadashi; Weir, William; Gardner, Malcolm; Pain, Arnab; Shiels, Brian; Hattori, Masahira; Nene, Vishvanath; Sugimoto, Chihiro

    2012-01-01

    We sequenced the genome of Theileria orientalis, a tick-borne apicomplexan protozoan parasite of cattle. The focus of this study was a comparative genome analysis of T. orientalis relative to other highly pathogenic Theileria species, T. parva and T. annulata. T. parva and T. annulata induce transformation of infected cells of lymphocyte or macrophage/monocyte lineages; in contrast, T. orientalis does not induce uncontrolled proliferation of infected leukocytes and multiplies predominantly within infected erythrocytes. While synteny across homologous chromosomes of the three Theileria species was found to be well conserved overall, subtelomeric structures were found to differ substantially, as T. orientalis lacks the large tandemly arrayed subtelomere-encoded variable secreted protein-encoding gene family. Moreover, expansion of particular gene families by gene duplication was found in the genomes of the two transforming Theileria species, most notably, the TashAT/TpHN and Tar/Tpr gene families. Gene families that are present only in T. parva and T. annulata and not in T. orientalis, Babesia bovis, or Plasmodium were also identified. Identification of differences between the genome sequences of Theileria species with different abilities to transform and immortalize bovine leukocytes will provide insight into proteins and mechanisms that have evolved to induce and regulate this process. The T. orientalis genome database is available at http://totdb.czc.hokudai.ac.jp/.

  10. Human leukocyte telomere length is associated with DNA methylation levels in multiple subtelomeric and imprinted loci.

    Science.gov (United States)

    Buxton, Jessica L; Suderman, Matthew; Pappas, Jane J; Borghol, Nada; McArdle, Wendy; Blakemore, Alexandra I F; Hertzman, Clyde; Power, Christine; Szyf, Moshe; Pembrey, Marcus

    2014-05-14

    In humans, leukocyte telomere length (LTL) is positively correlated with lifespan, and shorter LTL is associated with increased risk of age-related disease. In this study we tested for association between telomere length and methylated cytosine levels. Measurements of mean telomere length and DNA methylation at >450,000 CpG sites were obtained for both blood (N = 24) and EBV-transformed cell-line (N = 36) DNA samples from men aged 44-45 years. We identified 65 gene promoters enriched for CpG sites at which methylation levels are associated with leukocyte telomere length, and 36 gene promoters enriched for CpG sites at which methylation levels are associated with telomere length in DNA from EBV-transformed cell-lines. We observed significant enrichment of positively associated methylated CpG sites in subtelomeric loci (within 4 Mb of the telomere) (P telomere length, DNA methylation and gene expression in health and disease.

  11. [Gamma interferon induced in human leukocytes by phytohemagglutinin: its production and biological characteristics].

    Science.gov (United States)

    Danielescu, G; Maniu, H; Georgescu, T; Cajal, N

    1988-01-01

    Human gamma type interferon (IFN) preparations were obtained through phytohemagglutinin stimulation of leukocytes from the peripheral blood. Biological value of these preparations varied between 160 u and 800 u/ml, depending on leukocyte incubation medium, culture system and inductor conservation. The rising of the antiviral activity through association between gamma (3 u) and alpha (27 u) interferons was revealed by the virus quantity reduction (in this case the vesicular stomatitis virus was used) during a 24-hour multiplication cycle. The protection ensured by the mixture of the two types of interferon was about ten times higher than the additive effect of the two preparations. Study of the antiproliferative activity of a gamma interferon preparation was conducted on two human cell lines of tumoral origin (T-10 from a glioblastoma, and HEp-2) and revealed the difficulties to quantify precisely this property of the crude gamma interferon preparations.

  12. Dynamics of leukocytes and cytokines during experimentally induced Streptococcus uberis mastitis.

    Science.gov (United States)

    Rambeaud, M; Almeida, R A; Pighetti, G M; Oliver, S P

    2003-12-15

    Streptococcus uberis causes a significant proportion of clinical and subclinical intramammary infections (IMI) in lactating and non-lactating dairy cows. In spite of this, its pathogenesis is incompletely understood. A study was conducted to determine leukocyte and cytokine dynamics during experimentally induced S. uberis mastitis. Five Jersey and five Holstein cows were challenged via intramammary inoculation of S. uberis into two uninfected mammary glands. Sixteen of 20 challenged mammary glands developed clinical mastitis with peak clinical signs observed at 144 h. The number of S. uberis in milk increased (Pmastitis. Other mediators may be involved in initial leukocyte recruitment to the mammary gland, since increases in milk somatic cells occurred earlier than cytokine production.

  13. Necrotizing Ulcer After BCG Vaccination in a Girl With Leukocyte-adhesion Deficiency Type 1.

    Science.gov (United States)

    Kurosawa, Hidemitsu; Mizukami, Tomoyuki; Nunoi, Hiroyuki; Kato, Masaya; Sato, Yuya; Okuya, Mayuko; Fukushima, Keitaro; Katsuyama, Yoshihiko; Arisaka, Osamu

    2017-05-22

    Leukocyte-adhesion deficiency-1 is a recessively inherited disorder associated with recurrent bacterial infections, severe periodontitis, peripheral leukocytosis, and impaired wound healing. We diagnosed moderate-type leukocyte-adhesion deficiency-1 in a 7-year-old girl who developed a necrotizing ulcer after Bacillus Calmette-Guerin vaccination. The patient showed moderate expression of CD18 in neutrophils with a homozygous splice mutation with c.41_c.58+2dup20 of ITGB2 and experienced recurrent severe infections complicated with systemic lupus erythematosus. She received hematopoietic stem cell transplantation from a matched elder brother with heterozygous mutation of ITGB2, and has since remained free of infection and systemic lupus erythematosus symptoms without immunosuppression therapy.

  14. Two-dimensional electrophoretic analysis of human leukocyte proteins from patients with rheumatoid arthritis

    Energy Technology Data Exchange (ETDEWEB)

    Willard, K.E. (Argonne National Lab., IL); Thorsrud, A.K.; Munthe, E.; Jellum, E.

    1982-04-01

    Human leukocyte proteins from more than 150 patients with rheumatoid arthritis, together with age- and sex-matched controls, were analyzed by use of the ISO-DALT technique of two-dimensional polyacrylamide gel electrophoresis. Patients with ankylosing spondylitis, polymyalgia rheumatica, psoriatic arthritis, calcium tendinitis, post-infectious arthritis, and asymmetrical seronegative arthritis were also included as positive controls. Synthesis of several proteins, referred to by number as members of the Rheuma set, is shown to increase in the leukocyte preparations from patients with classical rheumatoid arthritis. Several of these proteins are specific to monocytes or granulocytes; others are of unknown cellular origin, but appear to be unique to rheumatoid arthritis. The Rheuma proteins appear to be indicators of disease activity, because their increased synthesis can be correlated with sedimentation rate and other clinical indices of rheumatoid disease activity.

  15. [The role of leukocytic factors in the preparation for and induction of labor].

    Science.gov (United States)

    Ermoshenko, B G; Shubich, M G

    2000-01-01

    Preparation for labour and its induction are controlled by an integrative but not single process. The primary stimulus that comprises this integrative hormonal prostaglandin-leukocytic mechanism is a suppression of local progesterone action with further inhibition of perivascular prostaglandine dehydrogenase and sharp increase of E2 and F2 prostaglandine content in uteroplacentary mechanism bed. Vasodilatory effect of prostaglandines along with synergication of interleukine 8 leads to leukocyte emigration and formation of infiltrates consisting of neutrophilic granulocytes and certain number of macrophages and mast cells. Hydrolytic enzymes released by neutrophilic granulocytes cause degradation of collagenic tissue in uterine cervix promoting its ripening. The factors mentioned also influence loosening of choriodecidual interface. Further amplification of effect of prostaglandine including those produced by neutrophilic granulocytes along with the other uteroconstrictors increase myometrium contractions which leads to labour induction.

  16. Visual and automated differential leukocyte counts. A comparison study of three instruments.

    Science.gov (United States)

    Rosvoll, R V; Mengason, A P; Smith, L; Patel, H J; Maynard, J; Connor, F

    1979-06-01

    The authors compared referee (senior author) microscopic counts, microscopic counts by several technologists, and counts obtained with two pattern-recognition leukocyte classifiers, (1) Larc and (2) Hematrak, and a cytochemical automated method for leukocyte counting, (3) Hermalog D, using samples from (1) a random patient population, (2) a selected abnormal patient population, and (3) healthy individuals. All instruments showed good accuracy and flagged abnormal results for review. Variability in pattern-recognition counts was found to be due mainly to the distribution of the cells on prepared blood smears. The Larc classifier was found to be very sensitive to minor alterations in the cytoplasm or nucleus, and rejected a number of slides. The Hemalog D showed the greatest precision. The method of preparation of slides would be a major decision factor in selecting one pattern-recognition instrument over another.

  17. Phosphatidylcholine reverses ethanol-induced increase in transepithelial endotoxin permeability and abolishes transepithelial leukocyte activation

    DEFF Research Database (Denmark)

    Mitscherling, K.; Volynets, V.; Parlesak, Alexandr

    2009-01-01

    BACKGROUND: Chronic alcohol abuse increases both intestinal bacterial overgrowth and intestinal permeability to macromolecules. Intestinal permeability of endotoxin, a component of the outer cell membrane of Gram-negative bacteria, plays a crucial role in the development of alcohol-induced liver...... disease (ALD). As impaired bile flow leads to endotoxemia and the bile component phosphatidylcholine (PC) is therapeutically active in ALD, we tested the hypothesis that conjugated primary bile salts (CPBS) and PC inhibit ethanol-enhanced transepithelial permeability of endotoxin and the subsequent...... transepithelial activation of human leukocytes. METHODS: For this purpose, we used a model in which intestinal epithelial cells (Caco-2) were basolaterally cocultivated with mononuclear leukocytes. Cells were challenged apically with endotoxin from Escherichia coli K12 and were incubated with or without...

  18. Phosphatidylcholine Reverses Ethanol-Induced Increase in Transepithelial Endotoxin Permeability and Abolishes Transepithelial Leukocyte Activation

    DEFF Research Database (Denmark)

    Mitzscherling, Katja; Volynets, Valentina; Parlesak, Alexandr

    2009-01-01

    Chronic alcohol abuse increases both intestinal bacterial overgrowth and intestinal permeability to macromolecules. Intestinal permeability of endotoxin, a component of the outer cell membrane of Gram-negative bacteria, plays a crucial role in the development of alcohol-induced liver disease (ALD......). As impaired bile flow leads to endotoxemia and the bile component phosphatidylcholine (PC) is therapeutically active in ALD, we tested the hypothesis that conjugated primary bile salts (CPBS) and PC inhibit ethanol-enhanced transepithelial permeability of endotoxin and the subsequent transepithelial...... activation of human leukocytes. For this purpose, we used a model in which intestinal epithelial cells (Caco-2) were basolaterally cocultivated with mononuclear leukocytes. Cells were challenged apically with endotoxin from Escherichia coli K12 and were incubated with or without the addition of CPBS (1.5 m...

  19. Roles of secretory leukocyte protease inhibitor amniotic membrane in oral wound healing

    Directory of Open Access Journals (Sweden)

    Elly Munadziroh

    2006-12-01

    Full Text Available Secretory Leukocyte Protease Inhibitor (SLPI is serine protease inhibitor. Secretory Leukocyte Protease Inhibitor is a protein found in secretions such as whole saliva, seminal fluid, cervical mucus, synovial fluid, breast milk, tears, and cerebral spinal fluid, as in secretions from the nose and bronchi, amniotic fluid and amniotic membrane etc. These findings demonstrate that SLPI function as a potent anti protease, anti inflammatory, bactericidal, antifungal, tissue repair, extra cellular synthesis. Impaired healing states are characterized by excessive proteolysis and often bacterial infection, leading to the hypothesis that SLPI may have a role in the process. The objectives of this article are to investigate the role of SLPI in oral inflammation and how it contributes to tissue repair in oral mucosa. The oral wound healing responses are impaired in the SLPI sufficient mice and matrix synthesis and collagen deposition are delayed. This study indicated that SLPI is a povital factor necessary for optimal wound healing.

  20. Effect of p-cymene on chemotaxis, phagocytosis and leukocyte behaviors

    Directory of Open Access Journals (Sweden)

    Raquel Kummer

    2015-03-01

    Full Text Available Summary. In this study, we investigated the effects of p-cymene (CYM in vitro and in vivo on leukocyte activity. In the cell viability assay, CYM (3, 10, 30, 90 g/mL had low cytotoxicity. In vitro chemotaxis revealed that CYM (3, 10, 30, 60 g/mL promoted a significant reduction of neutrophil migration toward fMLP, but not toward LTB4 stimulation. In carrageenan-induced peritonitis, CYM (100, 200, 400 mg/kg decreased the infiltration of peritoneal exudate leukocytes and of polymorphonuclear leukocytes. CYM pretreatment resulted in a significant decrease in the number of rolling (100, 200 mg/kg and in the number of adherent leukocytes (100, 200, 400 mg/kg to the perivascular tissue. The macrophage phagocytic index was increased significantly in concentrations of CYM (3, 10, 30 µg/ml. Treatment with CYM (10, 90 µg/ml also reduced TNF- levels but did not alter IL-10 levels in fMLP-stimulated neutrophils. In conclusion, CYM may be considered as a potential agent for treatment inflammatory injury, however, further studies are necessary to elucidate the anti-inflammatory mechanism.Industrial relevance. The CYM is a secondary metabolite, belongs to the class of monoterpene and is found in essential oils from plants, food and in several spices. It is an important intermediate used in pharmaceutical, flavor and aroma industries and for the production of fungicides. The investigation of the anti-inflammatory activity of this compound may be useful to development of new drug anti-inflammatory, on a commercial scale.Keywords. p-cymene; chemotaxis; in vivo microcirculation; phagocytosis; anti-inflammatory.

  1. High Phobic Anxiety Is Related to Lower Leukocyte Telomere Length in Women

    OpenAIRE

    Okereke, Olivia I.; Jennifer Prescott; Wong, Jason Y.Y.; Jiali Han; Rexrode, Kathryn M; Immaculata De Vivo

    2012-01-01

    BACKGROUND: Chronic psychological distress has been linked to shorter telomeres, an indication of accelerated aging. Yet, little is known about relations of anxiety to telomeres. We examined whether a typically chronic form of anxiety--phobic anxiety--is related to telomere length. METHODOLOGY/PRINCIPAL FINDINGS: Relative telomere lengths (RTLs) in peripheral blood leukocytes were measured by quantitative real-time polymerase chain reaction among 5,243 women (aged 42-69 years) who: were parti...

  2. Human Leukocytes Kill Brugia malayi Microfilariae Independently of DNA-Based Extracellular Trap Release

    OpenAIRE

    Ciaran J. McCoy; Reaves, Barbara J.; Giguère, Steeve; Coates, Ruby; Rada, Balázs; Wolstenholme, Adrian J.

    2017-01-01

    Background Wuchereria bancrofti, Brugia malayi and Brugia timori infect over 100 million people worldwide and are the causative agents of lymphatic filariasis. Some parasite carriers are amicrofilaremic whilst others facilitate mosquito-based disease transmission through blood-circulating microfilariae (Mf). Recent findings, obtained largely from animal model systems, suggest that polymorphonuclear leukocytes (PMNs) contribute to parasitic nematode-directed type 2 immune responses. When expos...

  3. Impaired leukocyte trafficking and skin inflammatory responses in hamsters lacking a functional circadian system.

    Science.gov (United States)

    Prendergast, Brian J; Cable, Erin J; Patel, Priyesh N; Pyter, Leah M; Onishi, Kenneth G; Stevenson, Tyler J; Ruby, Norman F; Bradley, Sean P

    2013-08-01

    The immune system is under strong circadian control, and circadian desynchrony is a risk factor for metabolic disorders, inflammatory responses and cancer. Signaling pathways that maintain circadian rhythms (CRs) in immune function in vivo, and the mechanisms by which circadian desynchrony impairs immune function, remain to be fully identified. These experiments tested the hypothesis that the hypothalamic circadian pacemaker in the suprachiasmatic nucleus (SCN) drives CRs in the immune system, using a non-invasive model of SCN circadian arrhythmia. Robust CRs in blood leukocyte trafficking, with a peak during the early light phase (ZT4) and nadir in the early dark phase (ZT18), were absent in arrhythmic hamsters, as were CRs in spleen clock gene (per1, bmal1) expression, indicating that a functional pacemaker in the SCN is required for the generation of CRs in leukocyte trafficking and for driving peripheral clocks in secondary lymphoid organs. Pinealectomy was without effect on CRs in leukocyte trafficking, but abolished CRs in spleen clock gene expression, indicating that nocturnal melatonin secretion is necessary for communicating circadian time information to the spleen. CRs in trafficking of antigen presenting cells (CD11c(+) dendritic cells) in the skin were abolished, and antigen-specific delayed-type hypersensitivity skin inflammatory responses were markedly impaired in arrhythmic hamsters. The SCN drives robust CRs in leukocyte trafficking and lymphoid clock gene expression; the latter of which is not expressed in the absence of melatonin. Robust entrainment of the circadian pacemaker provides a signal critical to diurnal rhythms in immunosurveilliance and optimal memory T-cell dependent immune responses. Copyright © 2013 Elsevier Inc. All rights reserved.

  4. Signal Inhibitory Receptor on Leukocytes-1 (SIRL-1), a potent regulator of neutrophil function

    OpenAIRE

    Avondt, K.

    2014-01-01

    In this thesis, we aimed to better understand the role of the immune inhibitory receptor Signal Inhibitory Receptor on Leukocytes-1 (SIRL-1) in neutrophil functions. Inhibitory receptors play a crucial role in tailoring immune responses. SIRL-1 belongs to a family of transmembrane receptors, the Ig superfamily, and is mainly expressed on granulocytes and monocytes. SIRL-1 contains two immunoreceptor tyrosine-based inhibitory motifs (ITIMs) in its cytoplasmic domain and recruits the phosphatas...

  5. Blood Level of Polymorphonuclear Neutrophil Leukocytes and Bronchial Hyperreactivity in Chronic Obstructive Pulmonary Disease

    OpenAIRE

    Cukic, Vesna

    2015-01-01

    Introduction: Polymorphonuclear neutrophil leukocytes (PMNL) have an important defensive role against various microorganisms and other agents, but by liberating various substances, first of all the superoxide anion (O 2¯), they can damage the bronchial mucosa and influence the development of bronchial inflammation which is the fundamental of bronchial hyperreactivity (BHR). Objective: to show the role of the PMNL for development and level of BHR in patients with chronic obstructive pulmonary ...

  6. Decline of semen quality and increase of leukocytes with cigarette smoking in infertile men

    Directory of Open Access Journals (Sweden)

    Zhi Hong Zhang

    2013-01-01

    Full Text Available Background: Previous researches about the effect of smoking on semen quality are contradictory, and the mechanism behind the harmful effect of smoking on semen quality still remains unclear until today. Objective: The objectives of this study are evaluation of the relationship between smoking and fertility, investigation of the effects of cigarette smoking on sperm parameters and detection of presence of leukocytes within the semen of idiopathic infertile men from Northeastern China. Materials and Methods: A retrospective study of 1512 infertile patients who visited affiliated hospitals of Jilin University from 2007-2010 were enrolled in this study. Patients were assigned into one non-smoking and one smoking group which was divided into mild, moderate and heavy subgroups. Sperm parameters (including leukocytes and sperm morphology analysis were performed using standard techniques. Results: Compared with non-smokers, smokers had a significant decrease in semen volumes (p=0.006, rapid progressive motility (p=0.002 and sperm viability (p=0.019; moreover, smokers had a significant increase in the levels of immotile sperms (p=0.005 and semen leukocytes (p=0.002; pH and sperm concentration were not statistically significant (p=0.789 and p=0.297 respectively. Sperm motion parameters were all lower in the smokers except for beat-cross frequency (Hz (BCF. Further, the percentage of normal morphology sperm was decreased significantly in smokers (p=0.003, the sperm morphology was worse with increasing degree of smoking. Conclusion: These findings suggest that smoking leads to a significant decline in semen quality and higher levels of leukocytes, thus smoking may affects the fertilization efficiency.

  7. TGF-b2 induction regulates invasiveness of Theileria-transformed leukocytes and disease susceptibility.

    Directory of Open Access Journals (Sweden)

    Marie Chaussepied

    Full Text Available Theileria parasites invade and transform bovine leukocytes causing either East Coast fever (T. parva, or tropical theileriosis (T. annulata. Susceptible animals usually die within weeks of infection, but indigenous infected cattle show markedly reduced pathology, suggesting that host genetic factors may cause disease susceptibility. Attenuated live vaccines are widely used to control tropical theileriosis and attenuation is associated with reduced invasiveness of infected macrophages in vitro. Disease pathogenesis is therefore linked to aggressive invasiveness, rather than uncontrolled proliferation of Theileria-infected leukocytes. We show that the invasive potential of Theileria-transformed leukocytes involves TGF-b signalling. Attenuated live vaccine lines express reduced TGF-b2 and their invasiveness can be rescued with exogenous TGF-b. Importantly, infected macrophages from disease susceptible Holstein-Friesian (HF cows express more TGF-b2 and traverse Matrigel with great efficiency compared to those from disease-resistant Sahiwal cattle. Thus, TGF-b2 levels correlate with disease susceptibility. Using fluorescence and time-lapse video microscopy we show that Theileria-infected, disease-susceptible HF macrophages exhibit increased actin dynamics in their lamellipodia and podosomal adhesion structures and develop more membrane blebs. TGF-b2-associated invasiveness in HF macrophages has a transcription-independent element that relies on cytoskeleton remodelling via activation of Rho kinase (ROCK. We propose that a TGF-b autocrine loop confers an amoeboid-like motility on Theileria-infected leukocytes, which combines with MMP-dependent motility to drive invasiveness and virulence.

  8. The internal architecture of leukocyte lipid body organelles captured by three-dimensional electron microscopy tomography.

    Directory of Open Access Journals (Sweden)

    Rossana C N Melo

    Full Text Available Lipid bodies (LBs, also known as lipid droplets, are complex organelles of all eukaryotic cells linked to a variety of biological functions as well as to the development of human diseases. In cells from the immune system, such as eosinophils, neutrophils and macrophages, LBs are rapidly formed in the cytoplasm in response to inflammatory and infectious diseases and are sites of synthesis of eicosanoid lipid mediators. However, little is known about the structural organization of these organelles. It is unclear whether leukocyte LBs contain a hydrophobic core of neutral lipids as found in lipid droplets from adipocytes and how diverse proteins, including enzymes involved in eicosanoid formation, incorporate into LBs. Here, leukocyte LB ultrastructure was studied in detail by conventional transmission electron microscopy (TEM, immunogold EM and electron tomography. By careful analysis of the two-dimensional ultrastructure of LBs from human blood eosinophils under different conditions, we identified membranous structures within LBs in both resting and activated cells. Cyclooxygenase, a membrane inserted protein that catalyzes the first step in prostaglandin synthesis, was localized throughout the internum of LBs. We used fully automated dual-axis electron tomography to study the three-dimensional architecture of LBs in high resolution. By tracking 4 nm-thick serial digital sections we found that leukocyte LBs enclose an intricate system of membranes within their "cores". After computational reconstruction, we showed that these membranes are organized as a network of tubules which resemble the endoplasmic reticulum (ER. Our findings explain how membrane-bound proteins interact and are spatially arranged within LB "cores" and support a model for LB formation by incorporating cytoplasmic membranes of the ER, instead of the conventional view that LBs emerge from the ER leaflets. This is important to understand the functional capabilities of leukocyte

  9. The internal architecture of leukocyte lipid body organelles captured by three-dimensional electron microscopy tomography.

    Science.gov (United States)

    Melo, Rossana C N; Paganoti, Guillherme F; Dvorak, Ann M; Weller, Peter F

    2013-01-01

    Lipid bodies (LBs), also known as lipid droplets, are complex organelles of all eukaryotic cells linked to a variety of biological functions as well as to the development of human diseases. In cells from the immune system, such as eosinophils, neutrophils and macrophages, LBs are rapidly formed in the cytoplasm in response to inflammatory and infectious diseases and are sites of synthesis of eicosanoid lipid mediators. However, little is known about the structural organization of these organelles. It is unclear whether leukocyte LBs contain a hydrophobic core of neutral lipids as found in lipid droplets from adipocytes and how diverse proteins, including enzymes involved in eicosanoid formation, incorporate into LBs. Here, leukocyte LB ultrastructure was studied in detail by conventional transmission electron microscopy (TEM), immunogold EM and electron tomography. By careful analysis of the two-dimensional ultrastructure of LBs from human blood eosinophils under different conditions, we identified membranous structures within LBs in both resting and activated cells. Cyclooxygenase, a membrane inserted protein that catalyzes the first step in prostaglandin synthesis, was localized throughout the internum of LBs. We used fully automated dual-axis electron tomography to study the three-dimensional architecture of LBs in high resolution. By tracking 4 nm-thick serial digital sections we found that leukocyte LBs enclose an intricate system of membranes within their "cores". After computational reconstruction, we showed that these membranes are organized as a network of tubules which resemble the endoplasmic reticulum (ER). Our findings explain how membrane-bound proteins interact and are spatially arranged within LB "cores" and support a model for LB formation by incorporating cytoplasmic membranes of the ER, instead of the conventional view that LBs emerge from the ER leaflets. This is important to understand the functional capabilities of leukocyte LBs in health and

  10. Concanavalin A as a probe for studying the mechanism of metabolic stimulation of leukocytes.

    Science.gov (United States)

    Romeo, D; Zabucchi, G; Jug, M; Miani, N; Soranzo, M R

    1975-01-01

    The disruption of the molecular organization of the plasma membrane of leukocytes by phagocytosable particles, or by agents such as surfactants, antibodies, phospholipase C, fatty acids and chemotactic factors, leads to a stimulation of the phagocyte oxidative metabolism. Concanavalin A (Con A) has been used as a tool to study the mechanism of this metabolic regulation. The binding of Con A to the surface of polymorphonuclear leukocytes (PMNL) or macrophages produces a rapid enhancement of oxygen uptake and glucose oxidation through the hexose monophosphate pathway (HMP). This is explained by an activation of the granular NADPH oxidase, the key enzyme in the metabolic stimulation. The effect of Con A is not due to endocytosed lectin, since Con A covalently coupled to large sepharose beads still acts as stimulant. The metabolic changes caused by Con A are reversible. If, after the onset of stimulation, sugars with high affinity for Con A are added to the leukocyte suspension, the activity of granular NADPH oxidase and the rate of respiration and glucose oxidation return to their resting values. The metabolic burst, while partially supressed by treatment of PMNL with iodoacetate, sodium flouride and cytochalasin B, is slightly increased by colchicine. Con A induces a selective release of granular enzymes (beta-glucuronidase, peroxidase, alkaline phosphatase) from PMNL, whereas no leakage of cytoplasmic enzymes is observed. The enzyme release is inhibited by iodoacetate and by drugs known to increase cell levels of cyclic AMP. Based on a current view of the mode of interaction between Con A and cell surfaces, a model of the metabolic disruption of leukocytes is presented.

  11. Porcine leukocyte cellular subsets sensitive to African swine fever virus in vitro.

    OpenAIRE

    1984-01-01

    African swine fever virus infected most, if not all, of the macrophages (monocytes) and ca. 4% of the polymorphonuclear leukocytes from porcine peripheral blood. B and T lymphocytes, either resting or stimulated with phytohemagglutinin, lipopolysaccharide, or pokeweed mitogen, were not susceptible to the virus. All of the mitogens used inhibited African swine fever multiplication in susceptible cells. The number of virus passages in vitro and the virulence degree of the virus did not affect t...

  12. Leukocyte scintigraphy: correlation of serial scintigraphic findings and clinical progression of inflammatory bowel disease

    Energy Technology Data Exchange (ETDEWEB)

    Ho, Y.; Kelly, M.J.; Kaliff, V. [Alfred Hospital, Prahan, VIC (Australia). Department of Nuclear Medicine

    1997-12-01

    Full text: This study was performed (a) to determine whether the clinical progress of individual patients with inflammatory bowel disease mirrored changes in leukocyte scans, and (b) to assess the reasons for significant discrepancies. Two nuclear medicine physicians reviewed 44 white cell scans in 20 consecutive patients (4 males, 18 females) who were referred for two or more leukocyte scans (using either the {sup 111}In Oxine or {sup 99m}Tc HMPAO labelling methods) by three gastroenterologists between 1 January 1992 and 1 June 1996. The sequential scanpairs (range 2-18 months apart) were classified by consensus reading as showing no change, more severe or less severe disease. Questionnaires were sent to the referring gastroenterologists to determine whether the overall clinical status of each patient was unchanged, better or worse in the interval between the two scans. There was complete agreement between clinical and scintigraphic assessment in 45% (10/22) of the study pairs. Review of responses of the three individual gastroenterologists showed a wide range of agreement (4/4, 4/5, 2/13). Review of data showed that most disagreement was based on subjective clinical assessment, and hence of uncertain significance. In two patients, however, potentially preventable false negative leukocyte scans occurred in patients with active proctitis. This may be overcome by rigorous attention to ensure complete emptying of radioactivity from the bladder when {sup 99m}Tc HMPAO is used. It was concluded that serial leukocyte scans add to clinical assessment but careful technique is needed to avoid false negative scans in the rectum

  13. Phagocytosis of Pseudomonas aeruginosa by polymorphonuclear leukocytes and monocytes: effect of cystic fibrosis serum.

    OpenAIRE

    Thomassen, M J; Demko, C A; Wood, R.E.; Sherman, J. M.

    1982-01-01

    It has been shown previously that serum from chronically infected patients with cystic fibrosis inhibits the phagocytosis of Pseudomonas aeruginosa by both normal and cystic fibrosis alveolar macrophages. In the present study, the ability of peripheral monocytes and polymorphonuclear leukocytes from normal volunteers and cystic fibrosis patients to phagocytize P. aeruginosa was shown not to be inhibited in the presence of serum from cystic fibrosis patients.

  14. Poly(ethyleneimine) modified filters for the removal of leukocytes from blood

    OpenAIRE

    Bruil, Anton; Oosterom, Hieke A.; Steneker, Ingeborg

    1993-01-01

    Polyurethane membrane filters and filters coated with poly(ethyleneimine) were used to investigate the influence of leukocyte adhesion during filtration. Treatment of the filters with an aqueous solution of 1% (w/v) poly(ethyleneimine) (PEI) led to the introduction of amine groups at the filter surfaces, as was confirmed by X-ray photoelectron spectroscopy. The modification procedure did not significantly change the porous structure in the filters, as was demonstrated by SEM and porometry. Us...

  15. Lipids and leukocytes in newborn umbilical vein blood, birth weight and maternal body mass index.

    Science.gov (United States)

    Brittos, T; de Souza, W B; Anschau, F; Pellanda, L

    2016-12-01

    Maternal obesity during pregnancy may influence fetal development and possibly predispose offspring to cardiovascular disease. The aim of the present study was to evaluate the relationship between maternal pre-pregnancy body mass index (BMI) and weight gain during pregnancy, and newborn birth weight, with lipid profile, high-sensitivity C-reactive protein (hs-CRP) and leukocyte in newborns. We performed a cross-sectional study of 245 mothers and their children. Blood was collected from the umbilical vein and assayed for lipid profile, hs-CRP and leukocyte count. Newborns average weight was 3241 g, total cholesterol 53.9 mg/dl, high-density lipoprotein cholesterol (HDL-c) 21.9 mg/dl, low-density lipoprotein cholesterol (LDL-c) 26.2 mg/dl, triglyceride 29.5 mg/dl and leukocytes 13,777/mm3. There was a direct correlation of pre-pregnancy BMI of overweight mothers with total cholesterol (r=0.220, P=0.037) and LDL-c (r=0.268, P=0.011) of newborns. Total cholesterol, LDL-c and HDL-c were higher in pre-term newborns (66.3±19.7, 35.9±14.6 and 25.2±7.7 mg/dl, respectively) that in full-term (52.4±13.1, 25.0±8.7 and 21.5±6.0 mg/dl), with P=0.001, 0.001 and 0.003, respectively. Leukocyte counts were higher in full-term newborns (14,268±3982/mm3) compared with pre-term (9792±2836/mm3, Pprogramming of the immune system.

  16. Age-dependent expression of osteochondrosis-related genes in equine leukocytes

    OpenAIRE

    Mendoza García, Luis; Piquemal, David; Lejeune, Jean-Philippe; Vander Heyden, Laurent; Noguier, F.; Bruno, R; Sandersen, Charlotte; Serteyn, Didier

    2015-01-01

    Introduction:  Osteochondrosis (OC) is a developmental disease in horses which has a significant impact on the horse's welfare and performance. The early disturbance in the process of endochondral ossification progresses to inflammatory and repair processes in older horses. Previously, differentially expressed genes in leukocytes of OC-affected horses have been identified. The aim of the present study is to detect age-related changes in these differentially expressed genes. Materials and Meth...

  17. F-actin-rich contractile endothelial pores prevent vascular leakage during leukocyte diapedesis through local RhoA signalling

    NARCIS (Netherlands)

    Heemskerk, N.; Schimmel, L.; Oort, C.; van Rijssel, J.; Yin, T.; Ma, B.; van Unen, J.; Pitter, B.; Huveneers, S.; Goedhart, J.; Wu, Y.; Montanez, E.; Woodfin, A.; van Buul, J.D.

    2016-01-01

    During immune surveillance and inflammation, leukocytes exit the vasculature through transient openings in the endothelium without causing plasma leakage. However, the exact mechanisms behind this intriguing phenomenon are still unknown. Here we report that maintenance of endothelial barrier integri

  18. Transfusion of leukocyte-depleted red blood cells is not a risk factor for nosocomial infections in critically ill children

    NARCIS (Netherlands)

    van der Wal, Judith; van Heerde, Marc; Markhorst, Dick G.; Kneyber, Martin C. J.

    Objectives: Transfusion of red blood cells is increasingly linked with adverse outcomes in critically ill children. We tested the hypothesis that leukocyte-depleted red blood cell transfusions were independently associated with increased development of bloodstream infections, ventilator-associated

  19. Increased prevalence of leukocytes and elevated cytokine levels in semen from Schistosoma haematobium-infected individuals.

    Science.gov (United States)

    Leutscher, Peter D C; Pedersen, Mette; Raharisolo, Clairette; Jensen, Jørgen Skov; Hoffmann, Steen; Lisse, Ida; Ostrowski, Sisse R; Reimert, Claus M; Mauclere, Philippe; Ullum, Henrik

    2005-05-15

    In this study, we investigated the seminal inflammatory response to egg infestation of the urogenital organs in 240 semen-donating men aged 15-49 years living in a Schistosoma haematobium-endemic area of Madagascar. In 29 subjects (12%) with excretion of > or =5 ova/ejaculate, leukocytospermia (>10(6) leukocytes/mL) and the presence of seminal lymphocytes and eosinophil leukocytes were each significantly more prevalent than in 74 subjects (31%) who were S. haematobium negative (Pinfection-negative subjects (Pinfection (STI) with Neisseria gonorrhoeae, Chlamydia trachomatis, Mycoplasma genitalium, and/or Trichomonas vaginalis did not act as a confounding factor for the observed associations. At follow-up, 6 months after systematic antischistosomiasis and STI syndrome treatment at baseline, the prevalence of seminal leukocytes decreased significantly among the previously seminal egg-positive subjects. The same tendency was observed for the posttreatment levels of cytokines. Numerous studies have already shown an association between STI-associated genital inflammation and human immunodeficiency virus (HIV) propagation. Therefore, the results of the present study suggest that male urogenital schistosomiasis may constitute a risk factor for HIV transmission, as a result of egg-induced inflammation in the semen-producing pelvic organs.

  20. Evaluation of Tc-99m leukocyte scan in the diagnosis of acute appendicitis

    Energy Technology Data Exchange (ETDEWEB)

    Butler, J.A.; Marcus, C.S.; Henneman, P.L.; Inkelis, S.H.; Wilson, S.E.

    1987-05-01

    A new /sup 99m/Tc Microlite leukocyte scan was performed in 38 patients to assess its value in the diagnosis of acute appendicitis. Autologous leukocytes are labeled with /sup 99m/Tc by inducing phagocytosis of /sup 99m/Tc albumin microcolloid particles. The advantages of this test over the standard indium-111 scan include superior imaging capability, a marked reduction (greater than 75%) in the radiation dose, and performance of the test including labeling, in less than 3 hr. Imaging is performed at 5-90 min postinjection of labeled cells. There were 19 male and 19 female patients with ages ranging from 10 to 80 years, in whom the diagnosis of appendicitis was indeterminate on clinical examination. Of the 13 of the 38 patients (34%) who came to surgery 12 had acute appendicitis. The WBC scan correctly identified 10 of the 12 cases of appendicitis. There were two false-negative studies. In the nonoperative group of 25 patients admitted for observation, 21 studies were reported as negative and four identified other sites of inflammation. All patients with a negative study have remained asymptomatic on follow-up. With a sensitivity of 83% (10/12) and a specificity of 100% (26/26) the /sup 99m/Tc leukocyte scan provides a rapid and highly accurate method for diagnosis of appendicitis in this preliminary study of patients with equivocal clinical exams.