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Sample records for bovine peripheral blood

  1. Phenotypic, ultra-structural and functional characterization of bovine peripheral blood dendritic cell subsets

    USDA-ARS?s Scientific Manuscript database

    Dendritic cells (DC) are multifunctional cells that bridge the gap between innate and adaptive immune systems. In bovine, significant information is lacking on the precise identity and role of peripheral blood DC subsets. In this study, we identify and characterize bovine peripheral blood DC subsets...

  2. Flow cytometric detection of bovine viral diarrhea virus in peripheral blood leukocytes of persistently infected cattle.

    PubMed Central

    Qvist, P; Aasted, B; Bloch, B; Meyling, A; Rønsholt, L; Houe, H

    1990-01-01

    Flow cytometry was investigated for detection of bovine viral diarrhea virus (BVDV) in peripheral blood mononuclear leukocytes of persistently infected cattle. The mononuclear leukocytes were purified by sedimentation in a gradient of Ficoll-Paque, fixed, permeabilized, and then labelled by indirect immunofluorescence using biotinylated immunoglobulins from a porcine antiserum to BVDV. Flow cytometric analysis of blood samples obtained from persistently infected cattle revealed virus in 3.0-21.0% (mean +/- SD, 11.2% +/- 6.4%) of the mononuclear leukocytes. Fluorescent cells were not observed in controls. Flow cytometric detection of BVDV in blood cells of persistently infected bovines is a rapid and objective technique which does not require cell culture facilities. PMID:2174298

  3. Phenotypic, Ultra-Structural, and Functional Characterization of Bovine Peripheral Blood Dendritic Cell Subsets

    PubMed Central

    Sei, Janet J.; Ochoa, Amanda S.; Bishop, Elizabeth; Barlow, John W.; Golde, William T.

    2014-01-01

    Dendritic cells (DC) are multi-functional cells that bridge the gap between innate and adaptive immune systems. In bovine, significant information is lacking on the precise identity and role of peripheral blood DC subsets. In this study, we identify and characterize bovine peripheral blood DC subsets directly ex vivo, without further in vitro manipulation. Multi-color flow cytometric analysis revealed that three DC subsets could be identified. Bovine plasmacytoid DC were phenotypically identified by a unique pattern of cell surface protein expression including CD4, exhibited an extensive endoplasmic reticulum and Golgi apparatus, efficiently internalized and degraded exogenous antigen, and were the only peripheral blood cells specialized in the production of type I IFN following activation with Toll-like receptor (TLR) agonists. Conventional DC were identified by expression of a different pattern of cell surface proteins including CD11c, MHC class II, and CD80, among others, the display of extensive dendritic protrusions on their plasma membrane, expression of very high levels of MHC class II and co-stimulatory molecules, efficient internalization and degradation of exogenous antigen, and ready production of detectable levels of TNF-alpha in response to TLR activation. Our investigations also revealed a third novel DC subset that may be a precursor of conventional DC that were MHC class II+ and CD11c−. These cells exhibited a smooth plasma membrane with a rounded nucleus, produced TNF-alpha in response to TLR-activation (albeit lower than CD11c+ DC), and were the least efficient in internalization/degradation of exogenous antigen. These studies define three bovine blood DC subsets with distinct phenotypic and functional characteristics which can be analyzed during immune responses to pathogens and vaccinations of cattle. PMID:25295753

  4. In vitro gene expression profile of bovine peripheral blood mononuclear cells in early Mycobacterium bovis infection

    PubMed Central

    CHENG, YAFEN; CHOU, CHUNG-HSI; TSAI, HSIANG-JUNG

    2015-01-01

    The intracellular parasite Mycobacterium bovis (M. bovis) causes tuberculosis in cattle and humans. Understanding the interactions between M. bovis and host cells is essential in developing tools for the prevention, detection, and treatment of M. bovis infection. Gene expression profiles provide a large amount of information regarding the molecular mechanisms underlying these interactions. The present study analyzed changes in gene expression in bovine peripheral blood mononuclear cells (PBMCs) at 0, 4 and 24 h following exposure to M. bovis. Using bovine whole-genome microarrays, a total of 420 genes were identified that exhibited significant alterations in expression (≥2-fold). Significantly enriched genes were identified using the Kyoto Encyclopedia of Genes and Genomes database, of which the highest differentially expressed genes were associated with the immune system, signal transduction, endocytosis, cellular transport, inflammation, and apoptosis. Of the genes associated with the immune system, 84.85% displayed downregulation. These findings support the view that M. bovis inhibits signaling pathways of antimycobacterial host defense in bovine PBMCs. These in vitro data demonstrated that molecular alterations underlying the pathogenesis of tuberculosis begin early, during the initial 24 h following M. bovis infection. PMID:26668602

  5. Interaction of Bovine Peripheral Blood Polymorphonuclear Cells and Leptospira Species; Innate Responses in the Natural Bovine Reservoir Host

    PubMed Central

    Wilson-Welder, Jennifer H.; Frank, Ami T.; Hornsby, Richard L.; Olsen, Steven C.; Alt, David P.

    2016-01-01

    Cattle are the reservoir hosts of Leptospira borgpetersenii serovar Hardjo, and can also be reservoir hosts of other Leptospira species such as L. kirschneri, and Leptospira interrogans. As a reservoir host, cattle shed Leptospira, infecting other animals, including humans. Previous studies with human and murine neutrophils have shown activation of neutrophil extracellular trap or NET formation, and upregulation of inflammatory mediators by neutrophils in the presence of Leptospira. Humans, companion animals and most widely studied models of Leptospirosis are of acute infection, hallmarked by systemic inflammatory response, neutrophilia, and septicemia. In contrast, cattle exhibit chronic infection with few outward clinical signs aside from reproductive failure. Taking into consideration that there is host species variation in innate immunity, especially in pathogen recognition and response, the interaction of bovine peripheral blood polymorphonuclear cells (PMNs) and several Leptospira strains was evaluated. Studies including bovine-adapted strains, human pathogen strains, a saprophyte and inactivated organisms. Incubation of PMNs with Leptospira did induce slight activation of neutrophil NETs, greater than unstimulated cells but less than the quantity from E. coli P4 stimulated PMNs. Very low but significant from non-stimulated, levels of reactive oxygen peroxides were produced in the presence of all Leptospira strains and E. coli P4. Similarly, significant levels of reactive nitrogen intermediaries (NO2) was produced from PMNs when incubated with the Leptospira strains and greater quantities in the presence of E. coli P4. PMNs incubated with Leptospira induced RNA transcripts of IL-1β, MIP-1α, and TNF-α, with greater amounts induced by live organisms when compared to heat-inactivated leptospires. Transcript for inflammatory cytokine IL-8 was also induced, at similar levels regardless of Leptospira strain or viability. However, incubation of Leptospira strains

  6. Molecular detection of bovine leukemia virus in peripheral blood of Iranian cattle, camel and sheep.

    PubMed

    Nekoei, S; Hafshejani, T Taktaz; Doosti, A; Khamesipour, F

    2015-01-01

    Bovine leukemia virus (BLV) is a deltaretrovirus which infects and induces proliferation of B-lymphocytes in the peripheral blood circulation and in lymphoid organs primarily of cattle, leading to leukemia/lymphoma. This study was carried out to investigate the presence of BLV in cattle, sheep and camels from the Chaharmahal va Bakhtiary and Isfahan provinces in Iran. A total of 874 blood samples collected from cattle, sheep and camels were used in this study to detect BLV using a nested-PCR. The results from this study indicated that 17.2% (n=874) of all blood samples collected were positive for BLV. The percentages of blood samples positive for BLV from cattle, sheep and camels were 22.1 (n=657), 5.3 (n=95) and 0 (n=122) respectively. The results from this study showed that BLV infected cattle and sheep. Camels seemed to be resistant to BLV infection. This study contributes to the nationwide effort to obtain baseline information on the prevalence of BLV, which will assist in planning the control strategy for the disease in Iran.

  7. Effects of polyunsaturated fatty acids on the proliferation of mitogen stimulated bovine peripheral blood mononuclear cells.

    PubMed

    Thanasak, J; Müller, K E; Dieleman, S J; Hoek, A; Noordhuizen, J P T M; Rutten, V P M G

    2005-04-08

    The present study aimed at analysis of the effects of polyunsaturated fatty acid (PUFA), linoleic acid (LA, C18:2n - 6) and linolenic acid (LNA, C18:3n - 3) on bovine peripheral blood mononuclear cells (PBMC) in vitro. Both mitogen (ConA)-induced proliferative lymphocyte responsiveness during 4 days of culture and eicosanoid (prostaglandin E(2) (PGE(2)) and leukotriene B(4) (LTB(4))) production during 36 h were determined in relation to the absence or presence of various concentrations of LA and LNA (0, 1, 5, 25, 125 and 250 microM). Mitogen-driven proliferative responses of lymphocytes tended to be uninfluenced in the presence of lower concentrations of LA, whereas significant inhibition was observed at the higher concentrations of LA (125 and 250 microM). However, increasing amounts of LNA did not affect the proliferation. ConA stimulation induced a clear PGE(2) response, which significantly decreased in the presence of 250 microM of LA. In addition, increasing amounts of LNA, but not LA, led to a significant decrease in LTB(4) levels. However, The production of LTB(4) did not alter due to mitogenic stimulation. In conclusion, the present study shows that bovine mononuclear cells may functionally be influenced by the presence of PUFA in their environment. Further studies need to be conducted to clarify in vivo consequences of these findings in a situation of PUFA enriched rations in ruminants.

  8. Hormonal modulation of phagocytosis and intracellular growth of Mycobacterium avium ss. paratuberculosis In bovine peripheral blood monocytes.

    PubMed

    Feola, R P; Collins, M T; Czuprynski, C J

    1999-01-01

    In this study, we evaluated the effects of several hormones (i.e. growth hormone, prolactin, vitamin D3, luteinizing hormone, oxytocin) on the phagocytosis and intracellular survival of Mycobacterium avium ss. paratuberculosis within bovine peripheral blood monocytes. Phagocytosis of M. avium ss. paratuberculosis declined in a dose-dependent manner when monocytes were exposed to increasing amounts of recombinant bovine growth hormone, with little phagocytosis occurring at a growth hormone concentration of 50 ng/ml. The other hormones tested had little effect on phagocytosis. Continuous exposure of bovine monocytes to bovine growth hormone (10 ng per ml) resulted in enhanced intracellular bacillary growth. This was detected within 3 days of monocyte infection, and resulted in a 1 Log10 greater number of M. avium ss. paratuberculosis in growth hormone treated, than control, monocytes at 12 days of infection. When monocytes were incubated with growth hormone for only the first 5 days of a 12 day incubation period, a further increase in bacillary multiplication was observed. A similar increase in bacillary multiplication was observed when M. avium ss. paratuberculosis monocytes were incubated with prolactin for the first 5 days of a 12 day incubation period. These data indicate that varying levels of growth hormone and prolactin can affect the intracellular multiplication of M. avium ss. paratuberculosis in bovine monocytes. Copyright 1999 Academic Press.

  9. Global Genetic Profiles of Gene Network Disruption in Bovine Peripheral Blood Mononuclear Cells Induced Bovine Leukemia Virus (BLV) Infection

    USDA-ARS?s Scientific Manuscript database

    Efficient nutrient assimilation into useful animal-derived products is the ultimate requirement for successful animal production. Infection in young growing animals can decrease energy and nutrient use required for growth rate by redirection of nutrients to support immune defense processes. Bovine l...

  10. In vitro permissivity of bovine peripheral blood mononuclear cells to bovine viral diarrhoea virus is dependent on the animal specific immune status.

    PubMed

    Lucchini, Barbara; Ponti, Wilma; Turin, Lauretta; Bronzo, Valerio; Scaccabarozzi, Licia; Luzzago, Camilla

    2012-04-01

    The in vitro permissivity to infection with homologous and heterologous bovine viral diarrhoea virus (BVDV) strains of bovine peripheral blood mononuclear cells (PBMCs) from eight naïve and eight BVDV-1b immune animals was studied. Four reference strains (BVDV-1a NADL, BVDV-1b NY-1, BVDV-2 125 and BVDV-2 890) were selected, based on genotype, prevalence and biotype. Virus neutralizing antibody titres were determined at bleeding and the viral loads were measured in PBMCs by end point titration in cell culture and by real-time PCR. PBMCs from both naïve and immune animals became infected by all BVDV strains tested, although virus titres were lower for immune heifers than naïve ones; the differences were significant for NADL (P<0.05) and 890 (P<0.001) strains. The in vitro model used in this study showed that PBMCs from immune animals are susceptible to re-infection with both homologous and heterologous BVDV strains, albeit at a lower extent than naïve cattle.

  11. In vitro replication activity of bovine viral diarrhea virus in an epithelial cell line and in bovine peripheral blood mononuclear cells.

    PubMed

    Turin, Lauretta; Lucchini, Barbara; Bronzo, Valerio; Luzzago, Camilla

    2012-11-01

    The present study focused on the in vitro infection of Madin-Darby bovine kidney (MDBK) cells and bovine peripheral blood mononuclear cells (PBMCs) from naÏve animals with non-cytopathic (ncp, BVDV-1b NY-1) and cytopathic (cp, BVDV-1a NADL) strains. Infections with 0.1 and 1 multiplicity of infections (MOI) and incubation times of 18 and 36 hr were compared. Twelve BVDV naÏve heifers were enrolled to collect PBMCs. The viral loads in MDBK cells and in PBMCs after in vitro infections were measured by real-time polymerase chain reaction (PCR) assays. The highest viral loads were measured at 1 MOI and 36 hr post infection in both cell systems and the lowest at 0.1 MOI and 18 hr with the exception of the cp strain NADL in PBMCs, for which the highest viral load was observed at 0.1 MOI and 36 hr. Viral load mean values were higher for the cp strain than the ncp strain irrespective of the extent of the infection period and MOI. The models of infection studied uncovered different replication activities respectively according to the biotype of virus, the cell substrate and the duration of infection. Replication tends to be higher in PBMCs, particularly at low MOIs and for the ncp strain.

  12. Road transportation stress promptly increases bovine peripheral blood absolute NK cell counts and cortisol levels.

    PubMed

    Ishizaki, Hiroshi; Kariya, Yoshihiro

    2010-06-01

    Livestock transportation effects on the number of circulating leukocytes have been reported. However, data related specifically to the relation between acute stress levels during transport and leukocyte differentiation, including lymphocyte subsets, are lacking. This study was undertaken to evaluate the distribution of peripheral blood leukocyte differential counts, CD25+ lymphocytes and NK cells in calves subjected to truck transportation on different road types. Healthy Japanese Black calves were divided into three treatments: 1) those moved around in a mountainous area (Group M); 2) those moved around on flatland (Group F); and 3) those that were not transported (control). The plasma cortisol levels in Group M increased during transport. The increase was significantly higher at the end of transport than in the controls (P<0.05); a slight increase was noted in Group F. Total leukocytes and the neutrophil to lymphocyte ratio in Group M were elevated with neutrophilia at 2 hr post-transport (P<0.05); the former levels remained higher than those in the controls for 4 hr. The numbers of lymphocytes, monocytes, eosinophils and CD25+ lymphocytes remained unchanged throughout the observations. The number of circulating NK cells in Group M increased during transport and peaked shortly after transport (P<0.05). Subsequent to these time points, the counts in Group F showed a trend toward elevation. The circulating NK cell counts were positively correlated with the plasma cortisol level during transport (M, r=0.755, P<0.0005; F, r=0.653; P<0.005). These results suggest that circulating NK cells might be more rapidly mobilized than other leukocytes. Therefore, they might reflect acute stress levels in calves during road transportation.

  13. Fuel feeds function: Energy balance and bovine peripheral blood mononuclear cell activation.

    PubMed

    Schwarm, A; Viergutz, T; Kuhla, B; Hammon, H M; Schweigel-Röntgen, M

    2013-01-01

    A general phenomenon in peripartum mammals is the breakdown of (acquired) immunity. The incidence of parasite load, disease and inflammation often rise during the specific energetically demanding time of pregnancy and lactation. In this period, blood leukocytes display decreased DNA synthesis in response to mitogens in vitro. Leukocyte activation, the phase of the cell cycle preceding the DNA synthetic phase has hardly been investigated, but the few studies suggest that leukocyte activation may also be impaired by the limited energy/nutrient availability. Leukocyte activation is characterized by manifold processes, thus, we used the cellular oxygen consumption rate (OCR) as a measure of ATP turnover to support all these processes. We hypothesized that the activation of peripheral blood mononuclear cells (PBMC) - in terms of oxygen consumed over basal levels after in vitro stimulation - is altered by energy balance around parturition. We studied peripartum high-yielding dairy cows because they undergo substantial fluctuations in energy intake, energy output and body fat mass. We established a fluorescence-based test strategy allowing for long-term (≥24h) quantification of O(2)-consumption and studied the peripartum period from 5 weeks ante partum to 5 weeks postpartum. In addition, we determined cellular lactate production, DNA/RNA synthesis and cell size and zoo-technical parameters such as animal energy intake and milk yield were assessed, as well as selected plasma parameters, e.g. glucose concentration. The basal OCR of PBMC from pregnant, non-lactating cows (n=6, -5 weeks ante partum) was 1.19±0.15 nmol min(-1) (10(7)cells)(-1) and increased to maximum levels of 2.54±0.49 nmol min(-1) (10(7)cells)(-1) in phytohemagglutinin (PHA)-stimulated PBMC. The basal OCR did not change over the peripartum period. Whereas the activation indices, herein defined as the PHA-induced 24h-increase of OCR above baseline, amounted to 1.1±0.3, 4.2±0.3, 4.1±1.1, 2.1±0.3, and

  14. Effect of bovine lactoferrin on functions of activated feline peripheral blood mononuclear cells during chronic feline immunodeficiency virus infection.

    PubMed

    Kobayashi, Saori; Sato, Reeko; Aoki, Takako; Omoe, Katsuhiko; Inanami, Osamu; Hankanga, Careen; Yamada, Yuichi; Tomizawa, Nobuyuki; Yasuda, Jun; Sasaki, Juso

    2008-05-01

    Feline immunodeficiency virus (FIV) infection is characterized by chronic overactivation of immune and inflammatory system, resulting in anergic state and dysfunction of immune cells. Lactoferrin (LF), a glycoprotein present in exocrine secretions and neutrophils, plays an important role in host defense system. Our previous study showed that oral administration of bovine LF (bLF) suppressed oral inflammation, improved the clinical symptoms and decreased serum gamma-globulin as a marker of inflammation in FIV-infected cats with intractable stomatitis. The anti-inflammatory effect was partly involved in regulation of neutrophil function by bLF. In this study, to clarify the relationship between anti-inflammatory effects of bLF and peripheral blood mononuclear cells (PBMC), we examined the effect of bLF on proliferation, cell cycle progression and cytokine expression in mitogen-activated PBMC. MTT [3-(4,5-dimethylthiazol-2-yl)- 2,5-diphenyl tetrazolium bromide] assay showed that bLF inhibited the concanavalin A (ConA)-induced cell proliferation in FIV-infected cats with the asymptomatic carrier and AIDS-related complex (ARC) phase. Bovine LF restored ConA-induced cell cycle progression and resulted in suppression of the induced apoptosis in feline PBMC. Real-time RT-PCR showed that bLF suppressed ConA-induced expression of interferon-gamma and interleukin-2 in cells of the ARC group regardless of the time of its addition to the medium. These results suggest the hypothesis that therapy with bLF may have the potential to improve and protect functions of overactivated lymphocytes by modulating the cell proliferation, cell cycle and cytokines expression in cats in terminal stage of FIV infection.

  15. Peripheral blood mononuclear cells from field cattle immune to bovine viral diarrhea virus (BVDV) are permissive in vitro to BVDV.

    PubMed

    Gupta, V; Mishra, N; Pateriya, A; Behera, S P; Rajukumar, K

    2014-01-01

    The aim of this study was to determine the in vitro permissivity of peripheral blood mononuclear cells (PBMCs) from bovine viral diarrhea virus (BVDV)-immune field cattle to homologous and heterologous BVDVs. PBMCs from seventeen BVDV-naïve and sixteen BVDV-immune animals were infected with noncytopathic BVDV-1 or BVDV-2. The immune status of cattle was indicated by the presence of virus neutralizing antibodies, while viral load of PBMCs was determined by real-time RT-PCR. The results revealed that the PBMCs from naïve or immune animals were permissive to either BVDV-1 or BVDV-2, but the viral load was significantly higher for the naïve than for the immune animals. Furthermore, the load of homologous virus in PBMCs from immune animals was lower than that of heterologous virus. Our results provide evidence that the PBMCs from BVDV-immune cattle in field are susceptible to reinfection with homologous or heterologous BVDV, albeit to a lower extent in the former case.

  16. Enzymatic Activities of Bovine Peripheral Blood Leukocytes and Milk Polymorphonuclear Neutrophils during Intramammary Inflammation Caused by Lipopolysaccharide

    PubMed Central

    Prin-Mathieu, C.; Le Roux, Y.; Faure, G. C.; Laurent, F.; Béné, M. C.; Moussaoui, F.

    2002-01-01

    Leukocytes are recruited from peripheral blood into milk as part of the inflammatory response to mastitis. However, excessive accumulation of inflammatory cells alters the quality of milk and the proteases produced by polymorphonuclear neutrophils (PMNs) and macrophages may lead to mammary tissue damage. To investigate PMN recruitment and the kinetics of their intracytoplasmic enzymes in inflammation, we generated mastitis in six cows by intramammary infusion of lipopolysaccharide (LPS). Clinical signs of acute mastitis were observed in all of the cows, and normal status was resumed by 316 h. Intracytoplasmic elastase, collagenase, and cathepsin activities were measured within live cells by flow cytometry in peripheral blood leukocytes and milk PMNs before and during the inflammatory process (at 10 time points between 4 and 316 h). The proportion of immature PMNs was appreciated by CD33 surface labeling measured in flow cytometry. Leukopenia was observed in the peripheral blood 4 h postinfusion, concomitant to an increase in somatic cell counts in milk. CD33+ PMNs were preferentially recruited from the peripheral blood to milk. Enzymatic activities were detected in PMNs, lymphocytes, and monocytes at levels depending on the cell type, sample nature, and time of collection. Milk PMNs had lower enzymatic activities than peripheral blood PMNs. This study showed that milk PMNs recruited during LPS-induced experimental mastitis have an immature phenotype and significantly lower enzymatic activities than peripheral blood PMNs. This suggests that CD33, an adhesion molecule, may be involved in the egress from blood to milk and that the enzymatic contents of PMNs are partly used during this process. PMID:12093678

  17. Decellularization of bovine anterior cruciate ligament tissues minimizes immunogenic reactions to alpha-gal epitopes by human peripheral blood mononuclear cells.

    PubMed

    Yoshida, Ryu; Vavken, Patrick; Murray, Martha M

    2012-10-01

    Rupture of ACL is a common injury. While the current surgical treatments are effective, many patients still suffer from precocious osteoarthritis, and there is an increasing interest in bioengineering approaches to improve ACL repair. Bovine collagen is a material currently in use for tissue engineering of ligaments. The alpha-gal epitopes found on bovine cells are a source of immunogenic stimulus for human cells. In this study, we wished to determine if those epitopes could be removed sufficiently to mitigate an immunogenic response using either a decellularization protocol or decellularization followed by alpha-galactosidase treatment. Bovine ACLs were treated with Triton-X, sodium deoxycholate, ribonuclease, and deoxyribonuclease to remove cells. A subset of the decellularized tissues was further treated with alpha-galactosidase. Human peripheral blood mononuclear cells (PBMCs) were exposed to untreated, decellularized, and alpha-galactosidase-treated tissues, and PBMC migration and IL-6 release were measured. PBMCs were significantly more attracted to untreated ACL compared to decellularized or alpha-galactosidase-treated tissue, but no difference was seen between the two treatment groups. PBMCs also released significantly more IL-6 when exposed to untreated tissue compared to decellularized ACL or alpha-galactosidase-treated ACL, but no difference was seen between the two treatment groups. Immunohistochemistry using anti-alpha-gal antibody detected the epitopes throughout the untreated ACL, but similar areas of reaction were not seen on decellularized or alpha-galactosidase-treated ACL. These results suggest that our decellularization protocol minimizes the immunogenic reactions of human PBMCs to bovine ACL tissue. Therefore, decellularized bovine ACL tissue may be a safe, effective biomaterial for ACL injury treatments.

  18. Effects of supplemental chromium on production of cytokines by mitogen-stimulated bovine peripheral blood mononuclear cells.

    PubMed

    Burton, J L; Nonnecke, B J; Dubeski, P L; Elsasser, T H; Mallard, B A

    1996-12-01

    This study determined whether supplementing the diets of dairy cows during the peripartum period with organic trivalent Cr influenced the capacity of their peripheral blood mononuclear cells to produce activation cytokines in response to stimulation with mitogens in vitro. Nine cows were fed 0.5 ppm of Cr/d per cow from 6 wk prepartum to 16 wk postpartum; 10 other periparturient cows served as unsupplemented controls. Mononuclear leukocytes, enriched from peripheral blood during wk 0, 2, 4, and 6 of lactation, were cultured with or without the T-lymphocyte mitogen, concanavalin A. Culture supernatants, harvested at 24, 48, or 72 h, were assayed for interleukin-2, interferon-gamma, and tumor necrosis factor-alpha. The cytokines were barely detectable in the supernatants from the unstimulated cultures, but supernatants from mitogen-stimulated cultures contained higher concentrations of each cytokine. For cows fed Cr, concentrations of all three cytokines in the culture supernatants of the mitogen-stimulated mononuclear cells decreased significantly relative to values for unsupplemented cows, particularly around peak lactation for the 24- and 48-h cultures. Theses results extended our previous observations and supported the hypothesis that organic Cr is immunomodulatory in high producing cows.

  19. Identification of three new bovine T-cell receptor delta variable gene subgroups expressed by peripheral blood T cells.

    PubMed

    Herzig, Carolyn T A; Blumerman, Seth L; Baldwin, Cynthia L

    2006-09-01

    To understand the biology of gammadelta T cells in ruminants, it is necessary to have a comprehensive picture of gammadelta T-cell receptor gene diversity and expression. In this study, three new subgroups of bovine T-cell receptor delta (TRD) variable genes were identified by RT-PCR and sequencing and homology with TRDV genes from other mammals determined. Previously unidentified TRDV subgroup genes described in this study include the bovine homologues of ovine TRDV2, TRDV3, and TRDV4 which were named accordingly. TRDV2 subgroup has two genes (TRDV2-1 and TRDV2-2) while we found the previously identified TRDV1 has at least eight genes corresponding to separate genomic sequences. Nucleotide and amino acid sequences for particular gene subgroups between cattle and sheep were more than 87% identical but identities among TRDV subgroups within a species were much less, with bovine TRDV4 having <45% identity to the other three bovine TRDV gene subgroups. Analysis of circulating bovine gammadelta T cells revealed that genes from all four TRDV subgroups were expressed in combination with TRDJ1, TRDJ3, and TRDC, although TRDV4 was the least represented, and all displayed a variety of CDR3 junctional lengths. Finally, some genes within the TRDV1, TRDV2, and TRDV3 subgroups recombined with TRAV incorporating TRAJs, suggesting dual use.

  20. Genetic characterization of bovine viral diarrhea virus strains in Beijing, China and innate immune responses of peripheral blood mononuclear cells in persistently infected dairy cattle.

    PubMed

    Weng, Xiao Gang; Song, Quan Jiang; Wu, Qiong; Liu, Ming Chao; Wang, Meng Ling; Wang, Jiu Feng

    2015-01-01

    To acquire epidemiological data on the bovine viral diarrhea virus (BVDV) and identify cattle persistently infected (PI) with this virus, 4,327 samples from Holstein dairy cows were screened over a four-year period in Beijing, China. Eighteen BVD viruses were isolated, 12 from PI cattle. Based on genetic analysis of their 5'-untranslated region (5'-UTR), the 18 isolates were assigned to subgenotype BVDV-1m, 1a, 1d, 1q, and 1b. To investigate the innate immune responses in the peripheral-blood mononuclear cells of PI cattle, the expression of Toll-like receptors (TLRs), RIG-I-like receptors, interferon-α (IFN-α), IFN-β, myxovirus (influenza virus) resistance 1 (MX1), and interferon stimulatory gene 15 (ISG15) was assessed by qPCR. When compared with healthy cattle, the expression of TLR-7, IFN-α, and IFN-β mRNA was downregulated, but the expression of MX1 and ISG-15 mRNA was upregulated in PI cattle. Immunoblotting analysis revealed that the expression of interferon regulatory factor 3 (IRF-3) and IRF-7 was lower in PI cattle than in healthy cattle. Thus, BVDV-1m and 1a are the predominant subgenotypes in the Beijing region, and the strains are highly divergent. Our findings also suggest that the TLR-7/IRF-7 signaling pathway plays a role in evasion of host restriction by BVDV.

  1. Eimeria bovis infection enhances adhesion of peripheral blood mononuclear cells to and their transmigration through an infected bovine endothelial cell monolayer in vitro.

    PubMed

    Taubert, Anja; Zahner, Horst; Hermosilla, Carlos

    2007-08-01

    The first schizogony of Eimeria bovis takes place in lymphatic endothelial cells of the ileum, resulting in the formation of macroschizonts within 2-3 weeks. In this study, we analyse early cellular immune responses to infected host cells on the basis of peripheral blood mononuclear cell (PBMC) adhesion on and transmigration through infected bovine umbilical vein endothelial cell (BUVEC) monolayers. Adhesion of PBMC was upregulated by an E. bovis infection. Most marked effects were observed 1 day p.i.; thereafter, PBMC adhesion declined reaching control levels from day 8 p.i. onward. CD8(+) T cells adhered more frequently to infected BUVEC (42%) than CD4(+) T cells (25%). About one third of attached PBMC were represented by gammadelta-TCR(+) T cells. Adhesion of T cells was not restricted to parasitised host cells, but occurred almost equally on non-infected BUVEC within the same monolayer. Furthermore, we found moderately enhanced levels of PBMC transmigration through infected BUVEC monolayers, in particular on day 2 p.i. The data presented here suggest that E. bovis infection of BUVEC induces endothelial cell-derived proinflammatory reactions, which appear suitable for the initiation of both adaptive and innate immune responses.

  2. Genetic characterization of bovine viral diarrhea virus strains in Beijing, China and innate immune responses of peripheral blood mononuclear cells in persistently infected dairy cattle

    PubMed Central

    Weng, Xiao Gang; Song, Quan Jiang; Wu, Qiong; Liu, Ming Chao; Wang, Meng Ling

    2015-01-01

    To acquire epidemiological data on the bovine viral diarrhea virus (BVDV) and identify cattle persistently infected (PI) with this virus, 4,327 samples from Holstein dairy cows were screened over a four-year period in Beijing, China. Eighteen BVD viruses were isolated, 12 from PI cattle. Based on genetic analysis of their 5'-untranslated region (5'-UTR), the 18 isolates were assigned to subgenotype BVDV-1m, 1a, 1d, 1q, and 1b. To investigate the innate immune responses in the peripheral-blood mononuclear cells of PI cattle, the expression of Toll-like receptors (TLRs), RIG-I-like receptors, interferon-α (IFN-α), IFN-β, myxovirus (influenza virus) resistance 1 (MX1), and interferon stimulatory gene 15 (ISG15) was assessed by qPCR. When compared with healthy cattle, the expression of TLR-7, IFN-α, and IFN-β mRNA was downregulated, but the expression of MX1 and ISG-15 mRNA was upregulated in PI cattle. Immunoblotting analysis revealed that the expression of interferon regulatory factor 3 (IRF-3) and IRF-7 was lower in PI cattle than in healthy cattle. Thus, BVDV-1m and 1a are the predominant subgenotypes in the Beijing region, and the strains are highly divergent. Our findings also suggest that the TLR-7/IRF-7 signaling pathway plays a role in evasion of host restriction by BVDV. PMID:26119170

  3. Relationship between quantity of IFNT estimated by IFN-stimulated gene expression in peripheral blood mononuclear cells and bovine embryonic mortality after AI or ET

    PubMed Central

    2012-01-01

    Background Interferon tau (IFNT), which is secreted into the uterine cavity during the maternal recognition period (MRP), is a key factor for establishment of pregnancy. The present study aims to clarify the relationship between the ability of a bovine conceptus to produce IFNT during the MRP and the conceptus's ability to establish pregnancy. Methods In the first experiment, IFNT (0, 500, or 1000 micrograms) was administered into the uterine horn ipsilateral to the CL 16 or 17 d after standing estrus, and mRNA levels of IFN-stimulated gene 15-kDa protein (ISG15) and Mx2 in peripheral blood mononuclear cells (PBMCs) were determined. In the second experiment, we investigated ISG15 mRNA expression in PBMCs during the MRP in cattle after either artificial insemination (AI) or embryo transfer (ET). Results Intrauterine administration of IFNT stimulated ISG15 and Mx2 gene expressions in PBMCs in cattle, and there was a positive correlation between the expressions of peripheral markers and the quantity of IFNT administered. In pregnant and normal interestrous interval (< 25 d) cattle (nIEI cattle), expression levels of the ISG15 gene showed similar patterns after AI and ET, and ISG15 mRNA expression was increased in pregnant cattle but unchanged in nIEI cattle. In contrast, ISG15 gene expression in extended interestrous interval (greater than or equal to 25 d) cattle (eIEI cattle) differed after ET compared with AI. In eIEI cattle after ET, ISG15 gene expression increased, such that the value on day 18 was intermediate between those of pregnant and nIEI cattle. In eIEI cattle after AI, ISG15 gene expression did not increase throughout the observation period. Conclusions The results of the current study indicate that the quantity of conceptus-derived IFNT can be estimated by measuring ISG15 mRNA levels in PBMCs from cattle. Using this approach, we demonstrate that ISG15 gene expression during the MRP in eIEI cattle differed after ET compared with AI. In addition, the

  4. Assessing transmission of Salmonella to bovine peripheral lymph nodes upon horn fly feeding

    USDA-ARS?s Scientific Manuscript database

    Blood-feeding ectoparasites are an economic burden to cattle production systems. Their role in mechanical dissemination of bacterial pathogens is also of significance. Biting arthropods are implicated in the transdermal transmission of Salmonella to bovine peripheral lymph nodes (PLNs), which may ul...

  5. PERIPHERAL BLOOD FILM - A REVIEW

    PubMed Central

    Adewoyin, AS; Nwogoh, B.

    2014-01-01

    The peripheral blood film (PBF) is a laboratory work-up that involves cytology of peripheral blood cells smeared on a slide. As basic as it is, PBF is invaluable in the characterization of various clinical diseases. This article highlights the basic science and art behind the PBF. It expounds its laboratory applications, clinical indications and interpretations in the light of various clinical diseases. Despite advances in haematology automation and application of molecular techniques, the PBF has remained a very important diagnostic test to the haematologist. A good quality smear, thorough examination and proper interpretation in line with patient's clinical state should be ensured by the haemato-pathologist. Clinicians should be abreast with its clinical utility and proper application of the reports in the management of patients. PMID:25960697

  6. Feeding soybean meal increases the blood level of isoflavones and reduces the steroidogenic capacity in bovine corpora lutea, without affecting peripheral progesterone concentrations.

    PubMed

    Cools, S; Van den Broeck, W; Vanhaecke, L; Heyerick, A; Bossaert, P; Hostens, M; Opsomer, G

    2014-01-30

    Thirty-three Holstein-Friesian cows were followed from 14 days pre partum until the fourth ovulation post partum. Housing conditions and basic ration were identical for all animals. Concentrates were individually supplemented according to the daily milk production level, using two different types of protein rich concentrates: soybean meal and rapeseed meal. Soybean and rapeseed meal are known to be respectively high and low in isoflavones. Cows were randomly divided into three groups and blocked for parity. Group I (n=11) was supplemented with soybean meal and acted as control group. Groups II (n=11) and III (n=11) were respectively supplemented with soybean and rapeseed meal and were subjected to a biopsy sampling of the corpus luteum at day 9 of the first three postpartal estrous cycles. Soybean meal supplementation to lactating dairy cows (1.72 kg on average) induced an increase in the blood concentration of equol, dihydrodaidzein, o-desmethylangolensin in both soy groups and resulted in a reduced area occupied by steroidogenic (P=0.012) and endothelial cells (P=0.0007) in the luteal biopsies. Blood concentrations of equol and glycitein were negatively correlated with the areas occupied by steroidogenic (r=-0.410 with P=0.0002, respectively r=-0.351 with P=0.008) and endothelial cells (r=-0.337 with P=0.01, respectively r=-0.233 with P=0.085) in the 3 first estrous cycles. The latter however did not affect the diestrous peripheral blood progesterone concentration. Copyright © 2014 Elsevier B.V. All rights reserved.

  7. Characterization of viral replication and the immune response in bison peripheral blood mononuclear cells following in vitro bovine viral diarrhea virus infection

    USDA-ARS?s Scientific Manuscript database

    Bovine viral diarrhea virus (BVDV) is a Pestivirus of the family Flaviviridae that has significant negative economic impact on beef and dairy production worldwide. In recent years, the North American bison industry has grown considerably with increases in the numbers of both wild and private herds. ...

  8. Bovine papillomavirus DNA in milk, blood, urine, semen, and spermatozoa of bovine papillomavirus-infected animals.

    PubMed

    Lindsey, C L; Almeida, M E; Vicari, C F; Carvalho, C; Yaguiu, A; Freitas, A C; Beçak, W; Stocco, R C

    2009-01-01

    Papillomavirus infection in bovines is associated with cutaneous papillomatosis on the hide, udders and other epithelial tissues, as well as in oral respiratory, alimentary and urinary tract mucosa. Bovine papillomavirus (BPV) is also considered the etiological agent of esophageal tumors and the malignant bladder tumors that characterize the clinical condition associated with chronic enzootic hematuria. After infective viral DNA was found in cattle blood and BPV1, 2 and 4 DNA in cattle reproductive and embryonic tissues, we looked for and found BPV DNA in blood, milk, urine, seminal fluid, and spermatozoa of BPV-infected animals. Peripheral blood lymphocyte cultures from BPV-infected animals had high rates of chromosome aberrations, including radial rearrangements that signal oncogenic potential and viral interaction with telomeric regions. The finding of BPV DNA in body fluids and tissues other than the epithelium demonstrates co-infection of other tissues or cell types by papillomavirus and shows the potential role of lymphocytes, seminal fluid and spermatozoa in BPV transmission. Our findings reinforce a peremptory need for prophylactic and therapeutic instruments to curtail this disease in bovine livestock.

  9. Stroma-free hemoglobin from bovine blood.

    PubMed

    Lima, Maria Celiana P; Andrade, Cristina T

    2007-01-01

    Isolation and purification of bovine hemoglobin (HbBv) was carried out after reaction of whole blood with carbon monoxide. Washing/centrifugation steps were used to eliminate leukocytes, platelets, and plasma proteins. Hypotonic media and ultrasound radiation were used to lyse red blood cells. Lyse by ultrasound was shown to lead to solutions at the highest concentrations in HbBv, and the least concentrations in major phospholipids contaminants. Additional purification procedures were performed to remove membrane proteins and phospholipids. In the first case, proteins were denatured by thermal treatment, and filtered. To eliminate phospholipids, liquid chromatography was used with strong anion exchangers. Purity of HbBv was evaluated by normal phase high performance liquid chromatography (HPLC), electrophoresis, and size-exclusion HPLC.

  10. Identification of highly active flocculant proteins in bovine blood

    USDA-ARS?s Scientific Manuscript database

    Bovine blood is an excellent flocculating agent, faster acting and as effective on a mass basis as polyacrylamide, the most widely utilized polymeric flocculant. To determine the molecular basis of flocculation activity, whole bovine blood (BB) and BB plasma were fractionated by size exclusion chro...

  11. Effects of cis-9,trans-11 and trans-10,cis-12 Conjugated Linoleic Acid, Linoleic Acid, Phytanic Acid and the Combination of Various Fatty Acids on Proliferation and Cytokine Expression of Bovine Peripheral Blood Mononuclear Cells

    PubMed Central

    Renner, Lydia; Kersten, Susanne; Duevel, Anna; Schuberth, Hans-Joachim; Dänicke, Sven

    2013-01-01

    Fatty acids may have an impact on immune functions, which is important in times of increased mobilization of body fat, e.g., around parturition. The aim of the present study was to investigate the effects of the CLA isomers cis-9,trans-11 and trans-10,cis-12, phytanic acid (PA), linoleic acid (LA) and a fatty acid (FA) mixture (containing 29.8% palmitic acid, 6.7% palmitoleic acid, 17.4% stearic acid and 46.1% oleic acid) on the proliferation of bovine blood mononuclear cells (PBMC) in vitro using alamar blue (AB) and 5-bromo-2′-deoxyuridine (BrdU) assay. Quantitative real time polymerase chain reaction analyses were performed to evaluate the expression of interleukin (IL)-4, IL-10, interferon (IFN)-γ, tumor necrosis factor (TNF)-α and peroxisome proliferator-activated receptor (PPAR)-γ in response to cis-9,trans-11 and LA. The IC50 values did not differ between the investigated FA, but there were differences within the proliferation in the response of these FA in a concentration range between 20 and 148 µM (e.g., increased proliferation after treatment with lower concentrations of LA). No differences occurred when different FA combinations were tested. ConA stimulation increased the expression of TNF-α and IFN-γ, whereas IL-10 decreased. In general, neither the baseline expression nor the ConA-stimulated mRNA expression of cytokines and PPAR-γ were affected by the FA. In conclusion, all FA inhibit the proliferation of PBMC dose dependently without significantly altering the induced cytokine spectrum of activated bovine PBMC. PMID:23857174

  12. Effects of cis-9,trans-11 and trans-10,cis-12 conjugated linoleic acid, linoleic acid, phytanic acid and the combination of various fatty acids on proliferation and cytokine expression of bovine peripheral blood mononuclear cells.

    PubMed

    Renner, Lydia; Kersten, Susanne; Duevel, Anna; Schuberth, Hans-Joachim; Dänicke, Sven

    2013-07-12

    Fatty acids may have an impact on immune functions, which is important in times of increased mobilization of body fat, e.g., around parturition. The aim of the present study was to investigate the effects of the CLA isomers cis-9,trans-11 and trans-10,cis-12, phytanic acid (PA), linoleic acid (LA) and a fatty acid (FA) mixture (containing 29.8% palmitic acid, 6.7% palmitoleic acid, 17.4% stearic acid and 46.1% oleic acid) on the proliferation of bovine blood mononuclear cells (PBMC) in vitro using alamar blue (AB) and 5-bromo-2'-deoxyuridine (BrdU) assay. Quantitative real time polymerase chain reaction analyses were performed to evaluate the expression of interleukin (IL)-4, IL-10, interferon (IFN)-γ, tumor necrosis factor (TNF)-α and peroxisome proliferator-activated receptor (PPAR)-γ in response to cis-9,trans-11 and LA. The IC50 values did not differ between the investigated FA, but there were differences within the proliferation in the response of these FA in a concentration range between 20 and 148 µM (e.g., increased proliferation after treatment with lower concentrations of LA). No differences occurred when different FA combinations were tested. ConA stimulation increased the expression of TNF-α and IFN-γ, whereas IL-10 decreased. In general, neither the baseline expression nor the ConA-stimulated mRNA expression of cytokines and PPAR-γ were affected by the FA. In conclusion, all FA inhibit the proliferation of PBMC dose dependently without significantly altering the induced cytokine spectrum of activated bovine PBMC.

  13. Automated microscopy system for peripheral blood cells

    NASA Astrophysics Data System (ADS)

    Boev, Sergei F.; Sazonov, Vladimir V.; Kozinets, Gennady I.; Pogorelov, Valery M.; Gusev, Alexander A.; Korobova, Farida V.; Vinogradov, Alexander G.; Verdenskaya, Natalya V.; Ivanova, Irina A.

    2000-11-01

    The report describes the instrument ASPBS (Automated Screening of Peripheral Blood Cells) designed for an automated analysis of dry blood smears. The instrument is based on computer microscopy and uses dry blood smears prepared according to the standard Romanovskii-Giemza procedure. In comparison with the well-known flow cytometry systems, our instrument provides more detailed information and offers an opporunity of visualizing final results. The basic performances of the instrument are given. Software of this instrument is based on digital image processing and image recognition procedures. It is pointed out that the instrument can be used as a fairly universal tool in scientific research, public demonstrations, in medical treatment, and in medical education. The principle used as the basis of the instrument appeared adequate for creating an instrument version serviceable even during space flights where standard manual procedures and flow cytometry systems fail. The benefit of the use of the instrument in clinical laboratories is described.

  14. Accumulation of L-type Bovine Prions in Peripheral Nerve Tissues

    PubMed Central

    Imamura, Morikazu; Matsuura, Yuichi; Masujin, Kentaro; Shimizu, Yoshihisa; Shu, Yujing; Kurachi, Megumi; Kasai, Kazuo; Murayama, Yuichi; Fukuda, Shigeo; Onoe, Sadao; Hagiwara, Ken’ichi; Yamakawa, Yoshio; Sata, Tetsutaro; Mohri, Shirou; Okada, Hiroyuki; Yokoyama, Takashi

    2010-01-01

    We recently reported the intraspecies transmission of L-type atypical bovine spongiform encephalopathy (BSE). To clarify the peripheral pathogenesis of L-type BSE, we studied prion distribution in nerve and lymphoid tissues obtained from experimentally challenged cattle. As with classical BSE prions, L-type BSE prions accumulated in central and peripheral nerve tissues. PMID:20587193

  15. Mobilization of peripheral blood stem cells.

    PubMed

    Arslan, Onder; Moog, Rainer

    2007-10-01

    The use of peripheral blood stem cells (PBSC) as a source of hematopoietic stem cells is steadily increasing and has nearly supplanted bone marrow transplantation. The present article reviews mobilization of PBSC as well as the side effects. Under steady state conditions less than 0.05% of the white blood cells (WBC) are CD34+ cells. Chemotherapy results in a 5-15-fold increase of PBSC. Combining chemotherapy and growth factors increases CD34+ cells up to 6% of WBC. In the allogeneic setting, granulocyte-colony stimulating factor (G-CSF) is used alone for PBSC mobilization. Several factors affect the mobilization of PBSC: age, gender, type of growth factor, dose of the growth factor and in the autologous setting, the patient's diagnosis, chemotherapy regimen and number of previous chemotherapy cycles or radiation. Muscle and bone pain are frequent adverse events in allogeneic stem cell mobilization but are usually tolerated with the use of analgesics. Spleen enlargement followed by rupture is a serious complication in allogeneic donors. Large volume apheresis (LVL) with a processed volume of more than 4-fold of the patient's blood volume can be used to increase the CD34+ yield in patients with low CD34+ pre-counts, resulting in higher yields of CD34+ cells for transplantation. Processing of more blood in LVL is achieved by an increase of the blood flow rate and an altered anticoagulation regimen with the occurrence of more citrate reactions.

  16. Purification of basophils from peripheral human blood.

    PubMed

    Falcone, Franco H; Gibbs, Bernhard F

    2014-01-01

    The purification of basophils from peripheral blood has represented a formidable challenge for researchers since they were discovered by Paul Ehrlich in 1879. From the first published attempts in the late 1960s, it took half a century to develop robust protocols able to provide sufficient numbers of pure, functionally unimpaired basophils. The existing protocols for basophil purification exploit those properties of basophils which distinguish them from other cell types such as their localization in blood, density, and the presence or absence of surface markers. Purification techniques have been used in various combinations and variations to achieve a common goal in mind: to obtain a pure population of human basophils in sufficient numbers for downstream studies. The arduous way leading up to the modern protocols is summarized in this historical retrospective. A fast protocol for purification of basophils to near homogeneity is also described.

  17. [Peripheral blood hematopoietic stem cell collection].

    PubMed

    Bojanić, Ines; Mazić, Sanja; Cepulić, Branka Golubić

    2009-01-01

    Summary. Peripheral blood hematopoietic stem cells (PBSC) have numerous advatages in comparison with traditionally used bone marrow. PBSC collection by leukapheresis procedure is simpler and better tolerated than bone marrow harvest. PBCS are mobilized by myelosupressive chemotherapy or/and hematopoietic growth factors. Leukapheresis product contains PBSC along with lineage commited progenitors and precursors which contribute to faster hematopoietic recovery. In "poor mobilizers" options are large-volume leukapheresis (LVL) procedure or second generation of mobilising agents (pegfilgrastim, CXCR4 receptor antagonists). Total blood volume is processed 2-3 times in standard procedure compared to more than 3 times in LVL. LVL yields significantly higher numbers of CD34+ cells. Adverse effects of leukapheresis are electrolyte disbalance (hypocalcemia) caused by citrat administration and risk of bleeding due to trobocytopenia and heparin administration. PBSC collection and product quality control are regulated by national and international standards and recommendations.

  18. Peripheral blood monocyte responses in periodontitis.

    PubMed

    Fokkema, S J

    2012-08-01

    Periodontitis results from the interaction of bacteria on the tooth surfaces and the host immune response. Although periodontal pathogens are essential for the initiation and progression of the disease, the tissue damage in periodontitis is primarily mediated by the host immune response. Differences in the susceptibility to the disease and in the clinal outcome of the therapy seem to be less dependent on genetics but more on lifestyle factors, like smoking, overweight, stress and nutrition. It has been shown that these lifestyle factors may modulate the immune response and therefore influence the initiation and progression of the disease. To study the host immune response, whole blood cell cultures (WBCC) stimulated with lipopolysaccharide (LPS) have been widely used and they specifically reflect the behaviour of monocytes. It has been shown that peripheral blood monocytes in LPS-stimulated WBCC from non-smoking periodontitis patients display a T-helper 2 (Th2)-promoting phenotype in comparison with controls. After periodontal therapy, this phenotype reversed and was comparable with controls. However, in smoking but treated patients, the Th2-promoting phenotype of monocytes still remained. Therefore, the aberrant phenotype of monocytes in the peripheral blood from periodontitis patients is likely to be a systemic response to exogenous and endogenous danger molecules released or induced by the periodontal infection or by smoking. It can be concluded that periodontal therapy in non-smoking periodontitis patients has beneficial health effects and that smoking cessation should be an integral part of the therapy as well for general health reasons as for the clinical outcome.

  19. Assessing Transmission of Salmonella to Bovine Peripheral Lymph Nodes upon Horn Fly Feeding.

    PubMed

    Olafson, Pia Untalan; Brown, Tyson R; Lohmeyer, Kimberly H; Harvey, Roger B; Nisbet, David J; Loneragan, Guy H; Edrington, Thomas S

    2016-07-01

    Biting arthropods are implicated in the transdermal transmission of Salmonella to bovine peripheral lymph nodes, and such contamination can contribute to increased Salmonella prevalence in processed beef. Since horn flies can acquire Salmonella and then excrete the bacteria in their feces, on-animal fly infestations were conducted in this study to assess whether horn flies have a role in this bacterial transmission. Three Salmonella serotypes were used to assess fly acquisition from and excretion onto cattle. The results indicated that flies can acquire Salmonella from the hide, as assessed by recovery from homogenates of surfacesterilized flies, and that Salmonella persists for at least 5 days in the fly. Fly fecal excreta serves as a bacterial contaminant on the hide, and the overall mean probable estimate of the quantity shed was ≈10(5) most probable number per fly cage area. In 5 days, no transmission of the bacteria to bovine peripheral lymph nodes was evident, prompting an assessment of the effects of prolonged horn fly feeding on transmission. Three groups of animals were infested with flies that had consumed a blood meal containing Salmonella Senftenberg. After 5 days, the study was either terminated or the flies were removed and the cages replenished with unfed flies either once or twice over the course of an 11- or 19-day fly exposure period, respectively. A microlancet-inoculated positive-control animal was included in each group for comparison. The impact of prolonged horn fly feeding was evident, as 8% of lymph nodes cultured were positive from the 5-day exposure, whereas 50 and 42% were positive from 11- and 19-day exposures, respectively. Higher concentrations of Salmonella were recovered from fly-infested animals than from the microlancet-inoculated control, likely a result of repeated inoculations over time by flies versus a single introduction. The data described provide new insights into the transmission dynamics of Salmonella in cattle

  20. The bovine peripheral-type benzodiazepine receptor: A receptor with low affinity for benzodiazepines

    SciTech Connect

    Parola, A.L.; Laird, H.E. II )

    1991-01-01

    The density of bovine peripheral-type benzodiazepine receptors (PBR) in four tissues was highest in adrenal cortex. The adrenal cortex PBR cofractionated with a mitochondrial membrane marker enzyme and could be solubilized with intact ligand binding properties using digitonin. The membrane bound and soluble mitochondrial receptors were pharmacologically characterized and showed the rank order of potency to inhibit ({sup 3}H)PK 11195 binding was PK 11195 > protoporphyrin IX > benzodiazepines. ({sup 3}H)PK 11195 binding to bovine adrenal mitochondria was unaffected by diethylpyrocarbonate, a histidine residue modifying reagent that decreased binding to rat liver mitochondria by 70%. ({sup 3}H)PK 14105 photolabeled the bovine PBR and the Mr was estimated under nondenaturing and denaturing conditions. These results demonstrate the bovine peripheral-type benzodiazepine receptor is pharmacologically and biochemically distinct from the rat receptor, but the receptor component photolabeled by an isoquinoline ligand has a similar molecular weight.

  1. Blood Thiamin Concentrations in Bovine Polioencephalomalacia

    PubMed Central

    Loew, F. M.; Dunlop, R. H.

    1972-01-01

    Thiamin concentrations were determined by the thiochrome method in blood from 16 cattle with polioencephalomalacia (PEM) or other apparent central nervous disorders. The mean blood total thiamin concentration of the five cattle with PEM (20.5 ± 5.1 µg/100 ml blood) was not significantly different from concentrations in the cattle with the other disorders, despite evidence of a thiamin diphosphate deficiency reported in cattle with PEM. PMID:4263913

  2. Human peripheral blood eosinophils induce angiogenesis.

    PubMed

    Puxeddu, Ilaria; Alian, Akram; Piliponsky, Adrian Martin; Ribatti, Domenico; Panet, Amos; Levi-Schaffer, Francesca

    2005-03-01

    Eosinophils play a crucial role in allergic reactions and asthma. They are also involved in responses against parasites, in autoimmune and neoplastic diseases, and in fibroses. There is increasing evidence that angiogenesis plays an important role in these processes. Since eosinophils are known to produce angiogenic mediators, we have hypothesized a direct contribution of these cells to angiogenesis. The effect of human peripheral blood eosinophil sonicates on rat aortic endothelial cell proliferation (in vitro), rat aorta sprouting (ex vivo) and angiogenesis in the chick embryo chorioallantoic membrane (in vivo) have been investigated. To determine whether eosinophil-derived vascular endothelial growth factor influences the eosinophil pro-angiogenic activity, eosinophil sonicates were incubated with anti-vascular endothelial growth factor antibodies and then added to the chorioallantoic membrane. Vascular endothelial growth factor mRNA expression and vascular endothelial growth factor receptor density on the endothelial cells were also evaluated. Eosinophils were found to enhance endothelial cell proliferation and to induce a strong angiogenic response both in the aorta rings and in the chorioallantoic membrane assays. Pre-incubation of eosinophil sonicates with anti-vascular endothelial growth factor antibodies partially reduced the angiogenic response of these cells in the chorioallantoic membrane. Eosinophils also increased vascular endothelial growth factor mRNA production on endothelial cells. Eosinophils are able to induce angiogenesis and this effect is partially mediated by their pre-formed vascular endothelial growth factor. This strongly suggests an important role of eosinophils in angiogenesis-associated diseases such as asthma.

  3. [Enterobacterial antigen in human peripheral blood lymphocytes].

    PubMed

    Faure-Fontenla, M A; García-Tamayo, F

    1989-11-01

    The following study has as prior history the research reports which have shown the existence of an antigenic tissue deposit in gram-negative enterobacteria. The antigens of the enterobacteria have also been found in the lymphocytic membranes and cytoplasm. Since intestinal lymphoid tissue cells can recirculate by means of the thoracic duct to the peripheral venous system, it was proposed that the circulating lymphocytes in healthy people could also contain small amounts of a common enterobacterial antigen. The study was carried out in 15 human venous blood samples, of which the lymphocytic population was separated to later be used in the preparation of 15 alcohol soluble extracts. This material was used for inhibiting the immuno-hemolysis assay in three occasions in order to show the presence of antigens shared by different enterobacterias, using as reference a fraction separated from the LPS of Escherichia coli 08. The results showed that the human lymphocytes also had antigenic determinants common to gram-negative bacteria.

  4. Enumeration of T cells, B cells and monocytes in the peripheral blood of normal and lymphocytotic cattle.

    PubMed Central

    Paul, P S; Senogles, D R; Muscoplat, C C; Johnson, D W

    1979-01-01

    The rosette-forming capacity of bovine peripheral blood lymphocytes (PBL) was determined with dextran and 2-aminoethylisothiouronium bromide (AET)-treated sheep erythrocytes (SRBC). Both dextran and AET-enhanced rosette formation; however, AET-treated SRBC detected a larger percentage of rosette-forming cells and thus was used in this study. The specificity of rosette formation by bovine thymus-derived (T) lymphocytes was shown by (1) demonstration of rosettes and surface-membrane immunoglobulins sIg) on different cells in PBL and nylon-wool fractionated lymphocyte populations and (2) rosette formation by a large percentage (83--90%) of thymocytes from three bovine foetuses and two 14-month-old heifers. A procedure was also developed to identify bovine monocytes by latex phagocytosis and 10--30% latex-ingesting cells were detected in PBL preparations isolated by Ficoll-Hypaque flotation. The frequency of sIg-bearing latex-ingesting, and sIg-bearing latex non-ingesting cells in bovine peripheral blood was also determined. These procedures were utilized to determine the distribution of T and bone-marrow derived (B) lymphocytes in peripheral blood of normal and lymphocytotic cattle. PBL from twenty normal cattle contained approximately 63% T and 11% B (sIg+ latex non-ingesting) lymphocytes. In peripheral blood of three cattle with persistent lymphocytosis, a prodromal stage of bovine leukaemia, the percentage of B cells was elevated approximately to 59% whereas T lymphocytes decreased to 35%, thus providing additional evidence that persistent lymphocytosis is a B-cell disease. PMID:312176

  5. Identification of highly active flocculant proteins in bovine blood.

    PubMed

    Piazza, George J; Nuñez, Alberto; Garcia, Rafael A

    2012-03-01

    Synthetic polymeric flocculants are used extensively for wastewater remediation, soil stabilization, and reduction in water leakage from unlined canals. Sources of highly active, inexpensive, renewable flocculants are needed to replace synthetic flocculants. High kaolin flocculant activity was documented for bovine blood (BB) and blood plasma with several anticoagulant treatments. BB serum also had high flocculant activity. To address the hypothesis that some blood proteins have strong flocculating activity, the BB proteins were separated by SEC. Then, the major proteins of the flocculant-active fractions were separated by SDS-PAGE. Identity of the major protein components was determined by tryptic digestion and peptide analysis by MALDI TOF MS. The sequence of selected peptides was confirmed using TOF/TOF-MS/MS fragmentation. Hemoglobin dimer (subunits α and β) was identified as the major protein component of the active fraction in BB; its high flocculation activity was confirmed by testing a commercial sample of hemoglobin. In the same manner, three proteins from blood plasma (fibrinogen, γ-globulin, α-2-macroglobulin) were found to be highly active flocculants, but bovine serum albumin, α-globulin, and β-globulin were not flocculants. On a mass basis, hemoglobin, γ-globulin, α-2-macroglobulin were as effective as anionic polyacrylamide (PAM), a widely used synthetic flocculant. The blood proteins acted faster than PAM, and unlike PAM, the blood proteins flocculants did not require calcium salts for their activity.

  6. Early Detection of NSCLC Using Stromal Markers in Peripheral Blood

    DTIC Science & Technology

    2015-09-01

    AWARD NUMBER: W81XWH-14-1-0263 TITLE: Early Detection of NSCLC Using Stromal Markers in Peripheral Blood PRINCIPAL INVESTIGATOR: Dingcheng...Sep 2014 - 31 Aug 2015 4. TITLE AND SUBTITLE Early Detection of NSCLC Using Stromal Markers in Peripheral Blood 5a. CONTRACT NUMBER W81XWH-14-1...SUPPLEMENTARY NOTES 14. ABSTRACT There is an immediate clinical need to develop a diagnostic biomarker for lung cancer in early stage. In this proposal

  7. Evaluation of a simple Theileria annulata culture protocol from experimentally infected bovine whole blood.

    PubMed

    Gharbi, M; Latrach, R; Sassi, L; Darghouth, M A

    2012-08-01

    We have evaluated a new simple technique using whole blood from experimentally infected cattle for the isolation and cultivation of Theileria annulata. The study was carried out on 20 Holstein-Frisian bovines that had been experimentally infected with a virulent lethal dose of Theileria annulata. This technique has been compared to the classical peripheral blood monocyte isolation with Ficoll carried out on 22 experimentally infected Holstein-Friesian calves. The effectiveness of the reference technique was estimated to 86.4%, whilst the effectiveness of the new technique was 100%. Moreover, this new technique leads to time and money saving estimated to € 3.06 per sample. It decreases the contamination risks by reducing the steps of sample manipulation.

  8. Effect of direct-fed microbial supplementation on the presence of Salmonella enterica in bovine peripheral lymph nodes

    USDA-ARS?s Scientific Manuscript database

    Introduction: Bovine peripheral lymph nodes (LN) contained within adipose trim, have been identified as a potential source of human exposure to Salmonella enterica, when incorporated into ground beef. How Salmonella gain entry to peripheral LN is a question yet to be answered, however recent survey...

  9. LEISHMANIA DONOVANI IN THE PERIPHERAL BLOOD

    PubMed Central

    Young, Charles W.; Van Sant, Helen M.

    1923-01-01

    1. The effect of human red cells and serum on the growth of Leishmania donovani has been studied and a blood culture method devised, based on these findings. 2. The distribution of blood platelets and of the different varieties of leucocytes in the strata of centrifugalized diluted blood has been investigated, and also the distribution of Leishman-Donovan bodies, extracellular and intracellular, in these strata. 3. The effect of treatment with antimony on the cultivability of Leishmania donavani has been studied. PMID:19868786

  10. Cardiac and peripheral blood similarities in the comparison of nordiazepam and bromazepam blood concentrations.

    PubMed

    Pos Pok, P-Rop; Haddouche, Djamel; Mauras, Michel; Kuhlmann, Erika; Burle, Joëlle; Salmon, Thierry; Berland, Emilie; Coiffait, P-Emmanuel; Viala, Alain

    2008-01-01

    Concomitant heart and peripheral blood determinations were performed on 40 fatal cases involving nordiazepam (20 cases) and bromazepam (20 cases). The heart blood concentration for the two drugs (588 ng/mL for nordiazepam and 802 ng/mL for bromazepam) does not differ from the corresponding peripheral blood concentration (587 ng/mL for nordiazepam and 883 ng/mL for bromazepam). The mean ratios for the heart and peripheral blood concentrations were 0.95 for nordiazepam and 0.86 for bromazepam. No postmortem redistribution was observed for these two benzodiazepines. The authors thus suggest that corresponding heart blood can be proposed in the quantitative analysis of these drugs when peripheral blood is unavailable. The present study also shows the stability of the two drugs after a year of storage.

  11. Peripheral blood findings associated with asymptomatic lead exposure

    SciTech Connect

    Day, C.M.; Tennant, F.S. Jr.

    1982-02-01

    This study was done to determine whether erythroid alterations can be found on a peripheral blood smear from an asymptomatic person exposure to excess atmospheric lead. Thirty healthy, asymptomatic adults who lived within five miles of a major Los Angeles, California freeway for five consecutive years were studied. Erythroid cytologic alterations-including-anisocytosis, poikilocytosis, polychromasia and basophilic stippling were statistically associated with increased free erythrocyte protoporphyrin levels. These findings indicate that erythroid alterations may be found on a peripheral blood smear prior to the development of clinical symptoms of lead intoxication.

  12. Peripheral blood stem cell mobilization failure.

    PubMed

    Kurnaz, Fatih; Kaynar, Leylagül

    2015-08-01

    Autologous hematopoietic stem cell transplantation (HSCT) is an important and often life saving treatment for many hematological malignancies and selected solid tumors. To rescue hematopoiesis after high-dose chemotherapy in autologous HSCT depends on maintaining sufficient stem cells. Hematopoietic stem cells and progenitor cells expressing CD34 in the BM are mobilized into the circulation with granulocyte-colony stimulating factor ± chemotherapy prior to autologous HSCT. One of the most important factors for success of autologous HSCT is hematopoietic stem cell (HSC) count. Minimum threshold for the engraftment of hematopoietic cells is accepted as 2 × 10(6) CD34 + cells/kg especially for platelet engraftment. Below this level it is defined as stem cell mobilization failure. There are several factors affecting stem cell mobilization: prior chemotherapy (such as fludarabine, melphalan, lenalidomide) and radiotherapy, age, type of disease, bone marrow cellularity. We tried to summarize the reasons of peripheral stem cell mobilization failure.

  13. Gene Expression Signature in Peripheral Blood Detects Thoracic Aortic Aneurysm

    PubMed Central

    Shiffman, Dov; Balasubramanian, Sriram; Iakoubova, Olga; Tranquilli, Maryann; Albornoz, Gonzalo; Blake, Julie; Mehmet, Necip N.; Ngadimo, Dewi; Poulter, Karen; Chan, Frances; Samaha, Raymond R.; Elefteriades, John A.

    2007-01-01

    Background Thoracic aortic aneurysm (TAA) is usually asymptomatic and associated with high mortality. Adverse clinical outcome of TAA is preventable by elective surgical repair; however, identifying at-risk individuals is difficult. We hypothesized that gene expression patterns in peripheral blood cells may correlate with TAA disease status. Our goal was to identify a distinct gene expression signature in peripheral blood that may identify individuals at risk for TAA. Methods and Findings Whole genome gene expression profiles from 94 peripheral blood samples (collected from 58 individuals with TAA and 36 controls) were analyzed. Significance Analysis of Microarray (SAM) identified potential signature genes characterizing TAA vs. normal, ascending vs. descending TAA, and sporadic vs. familial TAA. Using a training set containing 36 TAA patients and 25 controls, a 41-gene classification model was constructed for detecting TAA status and an overall accuracy of 78±6% was achieved. Testing this classifier on an independent validation set containing 22 TAA samples and 11 controls yielded an overall classification accuracy of 78%. These 41 classifier genes were further validated by TaqMan® real-time PCR assays. Classification based on the TaqMan® data replicated the microarray results and achieved 80% classification accuracy on the testing set. Conclusions This study identified informative gene expression signatures in peripheral blood cells that can characterize TAA status and subtypes of TAA. Moreover, a 41-gene classifier based on expression signature can identify TAA patients with high accuracy. The transcriptional programs in peripheral blood leading to the identification of these markers also provide insights into the mechanism of development of aortic aneurysms and highlight potential targets for therapeutic intervention. The classifier genes identified in this study, and validated by TaqMan® real-time PCR, define a set of promising potential diagnostic markers

  14. Assessment of Normal Variability in Peripheral Blood Gene Expression

    DOE PAGES

    Campbell, Catherine; Vernon, Suzanne D.; Karem, Kevin L.; ...

    2002-01-01

    Peripheral blood is representative of many systemic processes and is an ideal sample for expression profiling of diseases that have no known or accessible lesion. Peripheral blood is a complex mixture of cell types and some differences in peripheral blood gene expression may reflect the timing of sample collection rather than an underlying disease process. For this reason, it is important to assess study design factors that may cause variability in gene expression not related to what is being analyzed. Variation in the gene expression of circulating peripheral blood mononuclear cells (PBMCs) from three healthy volunteers sampled three times onemore » day each week for one month was examined for 1,176 genes printed on filter arrays. Less than 1% of the genes showed any variation in expression that was related to the time of collection, and none of the changes were noted in more than one individual. These results suggest that observed variation was due to experimental variability.« less

  15. A high-quality annotated transcriptome of swine peripheral blood

    USDA-ARS?s Scientific Manuscript database

    Background: High throughput gene expression profiling assays of peripheral blood are widely used in biomedicine, as well as in animal genetics and physiology research. Accurate, comprehensive, and precise interpretation of such high throughput assays relies on well-characterized reference genomes an...

  16. Peripheral blood stem cell transplantation for ischemic femoral head necrosis.

    PubMed

    Song, H-J; Lan, B-Sh; Cheng, B; Zhang, K-F; Yan, H-W; Wang, W-Zh; Gao, Z-Q

    2010-06-01

    Avascular necrosis of the femoral head (ANFH) is a highly mutilating disease. There is no effective way to treat femoral head ischemia. This study was designed to show the curative effects of peripheral blood stem cell transplantation to induce vascular regeneration and improve ischemic femoral head necrosis in rabbits. Twenty New Zealand white rabbits underwent ischemic femoral head necrosis in both hindlimbs using liquid-nitrogen refrigeration. One cohort of rats was intraperitoneally injected with granulocyte-specific colony-stimulating factor (250 microg/kg/d), and control animals received equivalent saline solution. The right side was used as the transplantation group and the left as the control. After separation of peripheral blood, a stem cell suspension was poured into the right femoral artery and saline solution into the left femoral artery. At 4 weeks after peripheral stem cell transplantation, standing ability and activity of the the transplanted right hindlimb were remarkably improved, but there were no obvious changes in the control limbs. The experimental rabbits underwent arteriography of bilateral femoral heads, which indicated increased and thickened blood supply to the transplanted right hindlimb compared with the left control. Peripheral blood stem cell transplantation improved ischemic femoral head necrosis.

  17. [Microangiopathy in preeclampsia: the usefulness of the peripheral blood smear].

    PubMed

    Duarte-Mote, Jesús; Espinosa-López, Rogelio F; Romero-Figueroa, Socorro; Lee Eng-Castro, Víctor E; Verduzco-Pineda, Julio; Calvo-Colindres, Jesús; Sánchez-Rojas, Graciela

    2012-01-01

    Endothelial dysfunction is a fundamental characteristic in the physiopathology of preeclampsia. Currently, a series of markers which explain endothelial dysfunction have been identified. The recognition of endothelial dysfunction has been used to realize an early diagnosis of preeclampsia, as soon as the classification of a possible prognosis. Nevertheless the detection of these markers is not accessible to the majority of hospitable centers that treat patients with preeclampsia. One indirect marker of endothelial dysfunction with a greater accessibility is the assessment of peripheral blood smear. Several studies had proved the presence of endothelial dysfunction by identification of red blood cells crenated in peripheral blood smear led us also to measure the impact in the evolution of the disease.

  18. Use of peripheral blood transcriptome biomarkers for epilepsy prediction

    PubMed Central

    Karsten, Stanislav L.; Kudo, Lili; Bragin, Anatol J.

    2011-01-01

    There are currently no predictive methods to identify patients who suffered an initial brain injury and are at high risk of developing chronic epilepsy. Consequently, treatments aimed at epilepsy prevention that would target the underlying epileptogenic process are neither available nor being developed. After a brain injury or any other initial precipitating event (IPE) to the development of epilepsy, pathological changes may occur in forms of inflammation, damage in the blood brain barrier, neuron loss, gliosis, axon sprouting, etc., in multiple brain areas. Recent studies provide connections between various kinds of brain pathology and alterations in the peripheral blood transcriptome. In this review we discuss the possibility of using peripheral blood transcriptome biomarkers for the detection of epileptogenesis and consequently, subjects at high risk of developing epilepsy. PMID:21419828

  19. Induction and identification of rabbit peripheral blood derived dendritic cells

    NASA Astrophysics Data System (ADS)

    Zhou, Jing; Yang, FuYuan; Chen, WenLi

    2012-03-01

    Purpose: To study a method of the induction of dendritic cells (DCs) from rabbit peripheral blood. Methods: Peripheral blood cells were removed from rabbit, filtered through nylon mesh. Peripheral blood mononuclear cells (PBMC) were isolated from the blood cells by Ficoll-Hypaque centrifugation (density of 1.077g/cm3).To obtain DCs, PBMC were cultured in RPMI1640 medium containing 10% fetal calf serum, 50U/mL penicillin and streptomycin, referred to subsequently as complete medium, at 37°C in 5% CO2 atmosphere for 4 hours. Nonadherent cells were aspirated, adherent cells were continued incubated in complete medium, supplemented with granulocyte/macrophage colony-stimulating factor (GM-CSF, 50ng/ml),and interleukin 4 (IL-4, 50ng/ml) for 9 days. Fluorescein labeled antibodies(anti-CD14, anti-HLA-DR, anti-CD86) were used to sign cells cultured for 3,6,9 days respectively, Then flow cytometry was performed. Results: Ratio of anti-HLA-DR and anti-CD86 labeled cells increased with induction time extension, in contrast with anti-CD14. Conclusion: Dendritic cells can be effectively induced by the method of this experiment, cell maturation status increased with induction time extension.

  20. Mouse cloning using a drop of peripheral blood.

    PubMed

    Kamimura, Satoshi; Inoue, Kimiko; Ogonuki, Narumi; Hirose, Michiko; Oikawa, Mami; Yo, Masahiro; Ohara, Osamu; Miyoshi, Hiroyuki; Ogura, Atsuo

    2013-08-01

    Somatic cell nuclear transfer (SCNT) is a unique technology that produces cloned animals from single cells. It is desirable from a practical viewpoint that donor cells can be collected noninvasively and used readily for nuclear transfer. The present study was undertaken to determine whether peripheral blood cells freshly collected from living mice could be used for SCNT. We collected a drop of peripheral blood (15-45 μl) from the tail of a donor. A nucleated cell (leukocyte) suspension was prepared by lysing the red blood cells. Following SCNT using randomly selected leukocyte nuclei, cloned offspring were born at a 2.8% birth rate. Fluorescence-activated cell sorting revealed that granulocytes/monocytes and lymphocytes could be roughly distinguished by their sizes, the former being significantly larger. We then cloned putative granulocytes/monocytes and lymphocytes separately and obtained 2.1% and 1.7% birth rates, respectively (P > 0.05). Because the use of lymphocyte nuclei inevitably results in the birth of offspring with DNA rearrangements, we applied granulocyte/monocyte cloning to two genetically modified strains and two recombinant inbred strains. Normal-looking offspring were obtained from all four strains tested. The present study clearly indicated that genetic copies of mice could be produced using a drop of peripheral blood from living donors. This strategy will be applied to the rescue of infertile founder animals or a "last-of-line" animal possessing invaluable genetic resources.

  1. A study of peripheral blood in hedgehogs in Turkey.

    PubMed

    Ozparlak, Haluk; Celik, Ilhami; Sur, Emrah; Ozaydin, Tuğba; Arslan, Atilla

    2011-09-01

    The aim of this study was to determine diameters of blood cells, differential counts of peripheral blood leukocytes, alpha-naphthyl acetate esterase (ANAE), acid phosphatase (ACP-ase) activity of some leukocyte types, and enzymatic positivity percentages of peripheral blood lymphocytes in two hedgehogs species, Hemiechinus auritus, the long-eared hedgehog, and Erinaceus concolor, the southern white-breasted hedgehog. Air-dried peripheral blood smears were stained with May-Grünwald-Giemsa stain. ANAE and ACP-ase were stained in glutaraldehyde-acetone-fixed smears. ANAE-positive lymphocytes displayed a dot-like positivity pattern characterized with 1-5 reddish brown cytoplasmic granules, whereas ACP-ase positive lymphocytes displayed a dot-like positivity pattern characterized with 1-3 pinkish cytoplasmic granules. Monocytes gave a diffuse and strong reaction while neutrophils displayed a weak positive reaction for ANAE and ACP-ase. No difference was observed in mean diameters of peripheral blood cells of these species. It was found that lymphocytes made up the majority (64.3% and 65.5%) of leukocytes, followed by neutrophils (23.9% and 23.3%), eosinophils (9.0% and 7.6%), monocytes (1.8% and 2.3%), and basophils (1.0% and 1.3%) in H. auritus and E. concolor, respectively. Mean ANAE positivity oflymphocytes was 36.6% and 51.3% and ACP-ase positivity was 32.1% and 37.5% for H. auritus and E. concolor, respectively. The ANAE positivity of lymphocytes in E. concolor was significantly (P < 0.05) higher than that of H. auritus.

  2. Management strategies for poor peripheral blood stem cell mobilization.

    PubMed

    Moog, Rainer

    2008-06-01

    Peripheral blood stem cells (PBSC) have nearly replaced bone marrow (BM) as the preferred source of hematopoietic rescue for patients undergoing high-dose chemotherapy. However, some patients fail to mobilize sufficient numbers of PBSC into the peripheral blood thereby putting high-dose chemotherapy at risk. The present article reviews mobilization of PBSC with a special focus on poor mobilizers. Under steady-state conditions less than 0.05% of the white blood cells (WBC) are CD34+ cells. Chemotherapy results in a 5-15-fold increase of PBSC. Combining chemotherapy and growth factors increases CD34+ cells up to 6% of WBC. Several factors affect the mobilization of PBSC: age, gender, type of growth factor, dose of the growth factor and in the autologous setting patient's diagnosis, chemotherapy regimen and number of previous chemotherapy cycles or radiation. Poor mobilizers are defined as patients with less than 10 CD34+ cells/mul in the peripheral blood during mobilization. Promising approaches for those patients rely on remobilization, use of high doses of granulocyte-colony stimulating factor (G-CSF), or the combination of G-CSF and granulocyte macrophage (GM)-CSF, which successfully mobilized the majority of poor mobilizing patients. New agents such as long lasting variants of G-CSF and CXCR4 antagonists are at the horizon and studied in clinical trials as mobilizing agents. Muscle and bone pain are frequent adverse events in stem cell mobilization but are usually tolerated under the use of analgesics. Large volume apheresis (LVL) with a processed volume of more than 4-fold patient's blood volume is an approach to increase the CD34+ yield in patients with low CD34+ pre-counts resulting in higher yields of CD34+ cells for transplantation. Processing of more blood in LVL is achieved by an increase of the blood flow rate and an altered anticoagulation regimen with the occurrence of more citrate reactions.

  3. A thermal peripheral blood flowmeter with contact force compensation

    NASA Astrophysics Data System (ADS)

    Sim, Jai Kyoung; Youn, Sechan; Cho, Young-Ho

    2012-12-01

    This paper presents a thermal peripheral blood flowmeter where a force sensor is integrated to compensate the blood flow measurement. Since blood flow is highly sensitive to the contact force between the sensor and skin, previous blood flowmeters needed to be fixed on the skin with a constant contact force. We integrate a force sensor with a thermal blood flowmeter to measure both blood flow and contact force simultaneously for force-compensated blood flow measurement. The blood flowmeter presented here is composed of a resistance temperature detector and a piezoresistive force sensor and was fabricated by surface and bulk micromachining techniques. In the experimental measurement, the blood flow linearly decreased with the contact force at the rate of 31.7% N-1. By using the measured compensation coefficient, the device showed a constant blood flow with the maximum difference of 6.4% over the contact force variation of 1-3 N, and otherwise showed the maximum difference of 75.0%. The present device is suitable for applications with portable biomedical instrumentation or air-conditioning systems for the estimation of human thermoregulation status.

  4. [239Pu and chromosomal aberrations in human peripheral blood lymphocytes].

    PubMed

    Okladnikova, N D; Osovets, S V; Kudriavtseva, T I

    2009-01-01

    The genome status in somatic cells was assessed using the chromosomal aberration (CA) test in peripheral blood lymphocytes from 194 plutonium workers exposed to occupational radiation mainly from low-transportable compounds of airborne 230Pu. Pu body burden at the time of cytogenetic study varied from values close to the method sensitivity to values multiply exceeding the permissible level. Standard (routine) methods of peripheral blood lymphocytes cultivation were applied. Chromatid- and chromosomal-type structural changes were estimated. Aberrations were estimated per 100 examined metaphase cells. The quantitative relationship between the CA frequency and Pu body burden and the absorbed dose to the lung was found. Mathematical processing of results was carried out based on the phenomenological model. The results were shown as theoretical and experimental curves. The threshold of the CA yield was 0.43 +/- 0.03 kBq (Pu body burden) and 6.12 +/- 1.20 cGy (absorbed dose to the lung).

  5. Reelin (RELN) DNA methylation in the peripheral blood of schizophrenia.

    PubMed

    Nabil Fikri, Rahim Mohd; Norlelawati, A Talib; Nour El-Huda, Abdul Rahim; Hanisah, Mohd Noor; Kartini, Abdullah; Norsidah, Kuzaifah; Nor Zamzila, Abdullah

    2017-05-01

    The epigenetic changes of RELN that are involved in the development of dopaminergic neurons may fit the developmental theory of schizophrenia. However, evidence regarding the association of RELN DNA methylation with schizophrenia is far from sufficient, as studies have only been conducted on a few limited brain samples. As DNA methylation in the peripheral blood may mirror the changes taking place in the brain, the use of peripheral blood for a DNA methylation study in schizophrenia is feasible due to the scarcity of brain samples. Therefore, the aim of our study was to examine the relationship of DNA methylation levels of RELN promoters with schizophrenia using genomic DNA derived from the peripheral blood of patients with the disorder. The case control studies consisted of 110 schizophrenia participants and 122 healthy controls who had been recruited from the same district. After bisufhite conversion, the methylation levels of the DNA samples were calculated based on their differences of the Cq values assayed using the highly sensitive real-time MethyLight TaqMan(®) procedure. A significantly higher level of methylation of the RELN promoter was found in patients with schizophrenia compared to controls (p = 0.005) and also in males compared with females (p = 0.004). Subsequently, the RELN expression of the methylated group was 25 fold less than that of the non-methylated group. Based upon the assumption of parallel methylation changes in the brain and peripheral blood, we concluded that RELN DNA methylation might contribute to the pathogenesis of schizophrenia. However, the definite effects of methylation on RELN function during development and also in adult life still require further elaboration. Copyright © 2017 Elsevier Ltd. All rights reserved.

  6. [Immunophenotypic characteristics of peripheral blood cells in normal elderly men].

    PubMed

    Wang, Ya-Zhe; Chang, Yan; Lu, Dan; Shi, Hong-Xia; Huang, Xiao-Jun; Liu, Yan-Rong

    2013-12-01

    This study was aimed to distinguish abnormal cells and to diagnose hematologic diseases through recognizing antigen expression pattern and percentage of peripheral blood cells in normal elderly men. Antigen expression of blast cells, granulocytes, monocytes, lymphocytes, nucleated red blood cells and plasma cells was detected by seven-color flow cytometry in a total of 88 peripheral blood samples from normal elderly men, aged median 82 years old, from 70 to 98 years. Groups were divided according to age, region and underlying diseases, and the percentages of different subgroup cells were examined to confirm whether the differences were significant or not. The results showed that the median proportion of CD34(+) blast cells in peripheral blood from normal elderly men were 0.017% (0.015%-0.020%), with high expression of HLA-DR, CD33, CD13 and CD117, low expression of myeloid antigens, such as CD15, CD11b and CD16, while lymphoid antigens were seldom positive, including CD7, CD19 and CD56. Dim-expression of CD38 was found in peripheral blood blast cells, CD38(dim)+/- cell percentage in blast cells was 61.36% ± 18.26%. In the differentiation and development of granulocytes, CD16(-), CD13(+) CD16(+) (intermediate) and CD16(+) (strong) CD13(+) cells appeared in sequence from immature to mature granulocytes, whose median proportions in nuclear cells were 0.04%, 0.30% and 61.30%, respectively. The percentages of immature monocytes, such as CD64(+) CD14(-) and HLA-DR(+) CD11b(-) cells, were from 0.00% to 0.10% and from 0.07% to 0.68%, separately. No significant differences were found between different subgroups (P > 0.05). It is concluded that the immunophenotypic characteristics and referential percentages of CD34(+) blast cells, granulocytes and monocytes with different development stages in peripheral blood from normal elderly men are recognized, which can help to discriminate abnormal cells.

  7. Detection of disseminated tumor cells in peripheral blood.

    PubMed

    Zieglschmid, V; Hollmann, C; Böcher, O

    2005-01-01

    Metastases are the major cause of cancer-related deaths in patients with solid epithelial malignancies, such as breast, colorectal and prostate carcinomas. Hematogenous spreading of tumor cells from a primary tumor can be considered as a crucial step in the metastasis cascade leading eventually to the formation of clinically manifest metastases. Consequently, as shown in recent studies, the detection of disseminated tumor cells in peripheral blood might be of clinical relevance with respect to individual patient prognosis and staging or monitoring of therapy. However, the rarity of disseminated tumor cells in peripheral blood renders the application of sensitive techniques mandatory for their detection. The emergence of highly sophisticated reverse transciptase-polymerase chain reaction (RT-PCR) assays, combining a preanalytical enrichment step with the assessment of multiple molecular tumor markers expressed in disseminated tumor cells, provides a powerful tool in detecting disseminated tumor cells with high sensitivity and specificity. This review will discuss currently used tumor markers as well as experimental means to enhance the sensitivity and specificity of RT-PCR assays to detect disseminated tumor cells in the peripheral blood of patients with breast, colorectal, and prostate cancers, and their clinical relevance assessed in recent studies.

  8. Peripheral markers of Alzheimer's disease: surveillance of white blood cells.

    PubMed

    Shad, Kaneez Fatima; Aghazadeh, Yashar; Ahmad, Sagheer; Kress, Bodo

    2013-08-01

    Inflammation is part of the complex biological response of vascular tissues to harmful stimuli, such as pathogens, damaged cells, or irritants. This is a mechanism of innate immunity, which may cause an increase in the number of monocytes and neutrophils circulating in the blood. Literature indicated that chronic inflammation might be a factor in developing neurological problems, including Alzheimer's, Parkinson's and other similar illnesses. Our main objective is to identify peripheral markers of Alzheimer's disease and for that purpose; we are looking at the profile of white blood cells focusing on monocytes, neutrophils, lymphocytes and basophils. Twenty-seven patients of Alzheimer's disease (AD), diagnosed by magnetic resonance imaging and neuropsychological tests were observed for their blood profile. Key observations during this study were that the levels of monocytes in the blood of the diagnosed AD patients were high irrespective of their age and sex. For those patients whose monocytes were in normal range their neutrophil levels were significantly high. Whereas blood levels of lymphocytes and basophils were found to be constantly low. Escalated levels of monocytes and neutrophils are hallmarks of chronic inflammation and may be precursor to Alzheimer's disease. A low lymphocyte count specifies that the body's resistance to fight infection is substantially reduced, whereas low basophil levels indicates their over utilization due to chronic allergic inflammatory condition. Future studies involved closer look at the cytokines produced by these white blood cells especially TNF IL-1, and IL-12, which are products of monocytes. Likewise, blood glucose and creatinine levels were high whereas calcium ions were low. Our studies indicated that white blood cells along with other inflammatory byproducts may act as peripheral markers for early diagnosis of Alzheimer's disease.

  9. Changes in CD34+ cell count in peripheral blood after whole blood donation.

    PubMed

    Pala, Cigdem; Mumcuoglu, Haluk; Kurnaz, Fatih; Sivgin, Serdar; Kaynar, Leylagul; Keklik, Muzaffer; Akyol, Gulsah; Koker, Yavuz; Eser, Bulent

    2013-10-01

    We aimed to investigate the change in the number of stem cells and white cells in the early period following blood donation. 22 male (71%) and 9 female (29%), 31 volunteers in total were included in the study. 450 ml of whole blood were collected from each of the volunteers for the donation. Complete blood counts were performed on the volunteers before and at 6 and 24h after the donation and CD34+ cell counts per ml of peripheral blood were measured by flow cytometry technique. There was a statistically significant increase in the number of CD34+ cells in the peripheral blood at 6h following blood donation (p<0.001). At 24h, however, there was a statistically significant decrease in the number of CD34+ cells, compared to 6h (p<0.001). There was a statistically significant increase in the number of leukocytes in the peripheral blood at 6h following blood donation (p<0.001). At 24h, there was a decrease in the number of leukocytes, which was statistically significant compared to 6h (p<0.001). When the difference in CD34+ cell and leukocytes counts before blood donation and at 24h after blood donation were compared, the results were not statistically significant. As the result of this study, a transient increase in the number of CD34+ cells in the peripheral blood after blood donation was demonstrated, with a decline in CD34+ cell counts back to levels prior to donation at 24h. Copyright © 2013 Elsevier Ltd. All rights reserved.

  10. Effect of parity on lymphocytes in peripheral blood and colostrum of healthy Holstein dairy cows

    PubMed Central

    Ohtsuka, Hiromichi; Terasawa, Sakiko; Watanabe, Chika; Kohiruimaki, Masayuki; Mukai, Machiko; Ando, Takaaki; Petrovski, Kiro R.; Morris, Stephen

    2010-01-01

    Investigation of the bovine systemic and mammary gland immune cells at calving might provide crucial information about the susceptibility of the mammary gland to infection. This study investigated the leukocyte population and cytokine mRNA levels in peripheral blood mononuclear cells (PBMCs) and colostrum mononuclear cells (CCs) obtained from healthy cows soon after calving. Fifty dairy cows that did not show clinical diseases were divided into 4 groups on the basis of parity: heifer (group 1, n = 10), 2nd calving (group 2, n = 11), 3rd calving (group 3, n = 14), and more than 3rd calving (group 4, n = 15). In the peripheral blood the numbers of CD3+TcR1-N12+, CD3+, CD4+, and major histocompatibility complex class II+CD14− lymphocytes were significantly higher in group 1 than in group 4, whereas in the colostrum the percentages of CD4+ and CD4+CD26+ lymphocytes and the CD4+/CD8+ ratio were significantly lower in group 1 than in group 4. There were no significant differences in the cytokine mRNA levels of PBMCs among the 4 groups; however, in the CCs the ratio of interferon gamma to interleukin 4 was significantly lower in group 1 than in group 3. These results suggest that the cellular immune function of PBMCs is lower, whereas mammary gland immune cells are more active, in cows with higher parity compared with heifers at calving. PMID:20592843

  11. Peripheral blood counts in workers exposed to synthetic fibres.

    PubMed

    Caciari, Tiziana; Casale, Teodorico; Loreti, Beatrice; Schifano, Maria P; Capozzella, Assunta; Scala, Barbara; De Sio, Simone; Tomei, Gianfranco; Rosati, Maria V; Tomei, Francesco

    2014-01-01

    Acrylonitrile is an intermediary with possible adverse health effects in the synthesis of organic products, such as acrylic fibres. This investigation was undertaken to determine the possible changes in the peripheral blood counts in workers of a polyacrylic fibres plant. The study involved 218 workers exposed to acrylonitrile at low doses and a control group of 200 unexposed workers. The chosen subjects underwent blood tests in order to check their haematological parameters. There were no statistically significant differences between the two groups in terms of the red blood cells, haemoglobin and total number of leukocytes. An increase in the neutrophils associated with a reduction of lymphocytes, both statistically significant, was observed. The authors hypothesized that the neutrophils are influenced by the exposure to acrylonitrile at low doses.

  12. Separation of hematopoietic stem cells from human peripheral blood through modified polyurethane foaming membranes.

    PubMed

    Higuchi, Akon; Sekiya, Mayu; Gomei, Yumiko; Sakurai, Masaru; Chen, Wen-Yih; Egashira, Satsuki; Matsuoka, Yuki

    2008-06-15

    Cell separation from peripheral blood was investigated using polyurethane (PU) foam membranes having 5.2 mum pore size and coated with Pluronic F127 or hyaluronic acid. The permeation ratio of hematopoietic stem cells (CD34(+) cells) and lymphocytes through the membranes was lower than for red blood cells and platelets. Adhered cells were detached from membrane surfaces using human serum albumin (HSA) solution after permeation of blood through the membranes, allowing isolation of CD34(+) cells in the permeate (recovery) solution. High-yield isolation of CD34(+) cells was achieved using Pluronic-coated membranes. This was because the Pluronic coating dissolved into the recovery solution at 4 degrees C, releasing adhered cells from the surfaces of the membranes during permeation of HSA solution through these membranes. Dextran and/or bovine serum albumin solutions were also evaluated for use as recovery solutions after blood permeation. A high recovery ratio of CD34(+) cells was achieved at 4 degrees C in a process using 20% dextran solution through PU membranes having carboxylic acid groups.

  13. Product characteristics and quality of bovine blood-enriched dried vegetable paste.

    PubMed

    Faria da Rocha, Silvia; Kappaun Rodrigues, Marla Cristina; Legemann Monte, Micheli; Quites Larrosa, Ana Paula; de Almeida Pinto, Luiz Antonio

    2014-12-01

    The aim of this study was to evaluate a preparation of vegetable paste with bovine blood in order to maximize the protein content using linear programming, and to analyze the product characteristics and quality of bovine blood-enriched vegetable paste dried in a spouted bed. The drying experiments were performed by evaluating the effects of inlet air temperature, paste flow rate and paste solids concentration on the dried product characteristics and quality (functional and nutritional properties). The vegetable paste enriched with bovine blood was a good source of protein (∼0.20 g g(-1) , dry basis), and the linear programming was adequate to select the constituents (carrot, onion, potato, kale, tomato, soybean oil and bovine blood) and optimize their quantities. The drying conditions of bovine blood-enriched vegetable paste in the spouted bed that gave the best product characteristics were an air temperature of 110 °C and a paste flow rate of 600 mL h(-1) with 0.07 g g(-1) solids concentration. The addition of bovine blood to vegetable paste by linear programming increased the protein content of the paste and improved its functional properties and digestibility. The powder obtained from the spouted bed drier showed suitable functional and nutritional properties and was also a good source of antioxidant compounds. © 2014 Society of Chemical Industry.

  14. Periodontal therapy alters gene expression of peripheral blood monocytes.

    PubMed

    Papapanou, Panos N; Sedaghatfar, Michael H; Demmer, Ryan T; Wolf, Dana L; Yang, Jun; Roth, Georg A; Celenti, Romanita; Belusko, Paul B; Lalla, Evanthia; Pavlidis, Paul

    2007-09-01

    We investigated the effects of periodontal therapy on gene expression of peripheral blood monocytes. Fifteen patients with periodontitis gave blood samples at four time points: 1 week before periodontal treatment (#1), at treatment initiation (baseline, #2), 6-week (#3) and 10-week post-baseline (#4). At baseline and 10 weeks, periodontal status was recorded and subgingival plaque samples were obtained. Periodontal therapy (periodontal surgery and extractions without adjunctive antibiotics) was completed within 6 weeks. At each time point, serum concentrations of 19 biomarkers were determined. Peripheral blood monocytes were purified, RNA was extracted, reverse-transcribed, labelled and hybridized with AffymetrixU133Plus2.0 chips. Expression profiles were analysed using linear random-effects models. Further analysis of gene ontology terms summarized the expression patterns into biologically relevant categories. Differential expression of selected genes was confirmed by real-time reverse transcriptase-polymerase chain reaction in a subset of patients. Treatment resulted in a substantial improvement in clinical periodontal status and reduction in the levels of several periodontal pathogens. Expression profiling over time revealed more than 11,000 probe sets differentially expressed at a false discovery rate of <0.05. Approximately 1/3 of the patients showed substantial changes in expression in genes relevant to innate immunity, apoptosis and cell signalling. The data suggest that periodontal therapy may alter monocytic gene expression in a manner consistent with a systemic anti-inflammatory effect.

  15. Periodontal therapy alters gene expression of peripheral blood monocytes

    PubMed Central

    Papapanou, Panos N.; Sedaghatfar, Michael H.; Demmer, Ryan T.; Wolf, Dana L.; Yang, Jun; Roth, Georg A.; Celenti, Romanita; Belusko, Paul B.; Lalla, Evanthia; Pavlidis, Paul

    2009-01-01

    Aims We investigated the effects of periodontal therapy on gene expression of peripheral blood monocytes. Methods Fifteen patients with periodontitis gave blood samples at four time points: 1 week before periodontal treatment (#1), at treatment initiation (baseline, #2), 6-week (#3) and 10-week post-baseline (#4). At baseline and 10 weeks, periodontal status was recorded and subgingival plaque samples were obtained. Periodontal therapy (periodontal surgery and extractions without adjunctive antibiotics) was completed within 6 weeks. At each time point, serum concentrations of 19 biomarkers were determined. Peripheral blood monocytes were purified, RNA was extracted, reverse-transcribed, labelled and hybridized with AffymetrixU133Plus2.0 chips. Expression profiles were analysed using linear random-effects models. Further analysis of gene ontology terms summarized the expression patterns into biologically relevant categories. Differential expression of selected genes was confirmed by real-time reverse transcriptase-polymerase chain reaction in a subset of patients. Results Treatment resulted in a substantial improvement in clinical periodontal status and reduction in the levels of several periodontal pathogens. Expression profiling over time revealed more than 11,000 probe sets differentially expressed at a false discovery rate of <0.05. Approximately 1/3 of the patients showed substantial changes in expression in genes relevant to innate immunity, apoptosis and cell signalling. Conclusions The data suggest that periodontal therapy may alter monocytic gene expression in a manner consistent with a systemic anti-inflammatory effect. PMID:17716309

  16. Peripheral blood eosinophilia has a favorable prognostic impact on transplant outcomes after allogeneic peripheral blood stem cell transplantation.

    PubMed

    Kim, Dong Hwan; Popradi, Gizelle; Xu, Wei; Gupta, Vikas; Kuruvilla, John; Wright, Janice; Messner, Hans A; Lipton, Jeffrey H

    2009-04-01

    Peripheral eosinophilia after allogeneic stem cell transplant (ASCT) may reflect the activation of the Th2 cytokine pathway. A retrospective analysis was performed to evaluate the impact of early- (before day 100: EEo) or late-onset (beyond day 100: LEo) eosinophilia (> or =0.5 x 10(9)/L in peripheral blood) on transplant outcomes after peripheral blood SCT (PBSCT) in 237 patients. The incidence of EEo and LEo was 43% at day 100 and 62% at 2 years, respectively. Compared with patients without LEo, improved transplant outcomes were observed in patients with LEo: better overall survival (OS; 86% versus 41%, P = 5 x 10(-11)), lower nonrelapse mortality (NRM; 10% versus 37%, P = 3 x 10(-6)), lower relapse incidence (11% versus 31%, P = 3 x 10(-5)), and higher GVHD-specific survival (GSS; 90% versus 64%, P = 1 x 10(-6)) were observed. In addition, similar finding was observed when transplant outcomes were analyzed according to the occurrence of eosinophilia at the onset of cGVHD. The multivariate analyses confirmed a favorable implication of LEo on OS, NRM, and GSS. LEo was associated with: (1) less severe chronic GVHD (cGVHD), (2) higher prevalence of autoantibodies, and (3) rapid lymphocyte count recovery after ASCT. In summary, the development of eosinophila after allogeneic PBSCT seemed to be a prognostic marker for improving transplant outcome.

  17. Peripheral blood smear image analysis: A comprehensive review.

    PubMed

    Mohammed, Emad A; Mohamed, Mostafa M A; Far, Behrouz H; Naugler, Christopher

    2014-01-01

    Peripheral blood smear image examination is a part of the routine work of every laboratory. The manual examination of these images is tedious, time-consuming and suffers from interobserver variation. This has motivated researchers to develop different algorithms and methods to automate peripheral blood smear image analysis. Image analysis itself consists of a sequence of steps consisting of image segmentation, features extraction and selection and pattern classification. The image segmentation step addresses the problem of extraction of the object or region of interest from the complicated peripheral blood smear image. Support vector machine (SVM) and artificial neural networks (ANNs) are two common approaches to image segmentation. Features extraction and selection aims to derive descriptive characteristics of the extracted object, which are similar within the same object class and different between different objects. This will facilitate the last step of the image analysis process: pattern classification. The goal of pattern classification is to assign a class to the selected features from a group of known classes. There are two types of classifier learning algorithms: supervised and unsupervised. Supervised learning algorithms predict the class of the object under test using training data of known classes. The training data have a predefined label for every class and the learning algorithm can utilize this data to predict the class of a test object. Unsupervised learning algorithms use unlabeled training data and divide them into groups using similarity measurements. Unsupervised learning algorithms predict the group to which a new test object belong to, based on the training data without giving an explicit class to that object. ANN, SVM, decision tree and K-nearest neighbor are possible approaches to classification algorithms. Increased discrimination may be obtained by combining several classifiers together.

  18. Modeled microgravity inhibits apoptosis in peripheral blood lymphocytes

    NASA Technical Reports Server (NTRS)

    Risin, D.; Pellis, N. R.; McIntire, L. V. (Principal Investigator)

    2001-01-01

    Microgravity interferes with numerous lymphocyte functions (expression of cell surface molecules, locomotion, polyclonal and antigen-specific activation, and the protein kinase C activity in signal transduction). The latter suggests that gravity may also affect programmed cell death (PCD) in lymphocyte populations. To test this hypothesis, we investigated spontaneous, activation- and radiation-induced PCD in peripheral blood mononuclear cells exposed to modeled microgravity (MMG) using a rotating cell culture system. The results showed significant inhibition of radiation- and activation-induced apoptosis in MMG and provide insights into the potential mechanisms of this phenomenon.

  19. Modeled Microgravity Inhibits Apoptosis in Peripheral Blood Lymphocytes

    NASA Technical Reports Server (NTRS)

    Risin, Diana; Pellis, Neal R.

    2000-01-01

    Microgravity interferes with numerous lymphocyte functions (expression of cell surface molecules, locomotion, polyclonal and antigen-specific activation, and the protein kinase C activity in signal transduction). The latter suggests that gravity may also affect programmed cell death (PCD) in lymphocyte populations. To test this hypothesis, we investigated spontaneous, activation- and radiation-induced PCD in peripheral blood mononuclear cells (PBMC) exposed to modeled microgravity using a rotating cell culture system. The results showed significant inhibition of radiation- and activation-induced apoptosis in modeled microgravity and provide insights into the potential mechanisms of this phenomenon.

  20. [Autologous transplantation of peripheral blood stem cells in malignant lymphomas].

    PubMed

    Sánchez Luceros, A; Koziner, B

    1995-01-01

    The administration of high dose chemotherapy and or radiotherapy with autologous hematopoietic rescue has become a treatment modality with increasing number of indications in a variety of malignant conditions. Improvements in the conditioning regimens and supportive measures used, as well as a more refined patient selection based on prognostic factors, have resulted in progressively better results. The availability of precursor cells from peripheral blood has allowed a faster restoration of hematopoiesis, decreasing the period and intensity of myelosuppression. The following revision gives an updated image of the accumulated experience with this mode of support in malignant lymphomas.

  1. Oropouche virus is detected in peripheral blood leukocytes from patients.

    PubMed

    de Souza Luna, Luciano Kleber; Rodrigues, Alcir Humberto; Santos, Rodrigo Ivo Marques; Sesti-Costa, Renata; Criado, Miriã Ferreira; Martins, Ronaldo B; Silva, Maria Lúcia; Delcaro, Luana Sella; Proença-Modena, Jose Luiz; Figueiredo, Luiz Tadeu Moraes; Acrani, Gustavo Olszanski; Arruda, Eurico

    2017-06-01

    Oropouche virus (OROV) is a frequent cause of arboviral febrile disease in the Amazon. The present report describes studies done in two patients, one of them; the first OROV human case acquired outside of the Amazon, which have revealed for the first time the presence of OROV in peripheral blood leukocytes. This novel finding raises important issues regarding pathogenesis of human infections and may offer a new tool, for the rapid diagnosis of this neglected infection. J. Med. Virol. 89:1108-1111, 2017. © 2017 Wiley Periodicals, Inc. © 2017 Wiley Periodicals, Inc.

  2. Modeled microgravity inhibits apoptosis in peripheral blood lymphocytes

    NASA Technical Reports Server (NTRS)

    Risin, D.; Pellis, N. R.; McIntire, L. V. (Principal Investigator)

    2001-01-01

    Microgravity interferes with numerous lymphocyte functions (expression of cell surface molecules, locomotion, polyclonal and antigen-specific activation, and the protein kinase C activity in signal transduction). The latter suggests that gravity may also affect programmed cell death (PCD) in lymphocyte populations. To test this hypothesis, we investigated spontaneous, activation- and radiation-induced PCD in peripheral blood mononuclear cells exposed to modeled microgravity (MMG) using a rotating cell culture system. The results showed significant inhibition of radiation- and activation-induced apoptosis in MMG and provide insights into the potential mechanisms of this phenomenon.

  3. Assessment of radio-frequency electromagnetic radiation by the micronucleus test in bovine peripheral erythrocytes.

    PubMed

    Balode, Z

    1996-02-02

    Previous bioindicative studies in the Skrunda Radio Location Station area have focused on the somatic influence of electromagnetic radiation on plants, but it is also important to study genetic effects. We have chosen cows as test animals for cytogenetical evaluation because they live in the same general exposure area as humans, are confined to specific locations and are chronically exposed to radiation. Blood samples were obtained from female Latvian Brown cows from a farm close to and in front of the Skrunda Radar and from cows in a control area. A simplified alternative to the Schiff method of DNA staining for identification of micronuclei in peripheral erythrocytes was applied. Microscopically, micronuclei in peripheral blood erythrocytes were round in shape and exhibited a strong red colour. They are easily detectable as the only coloured bodies in the uncoloured erythrocytes. From each individual animal 2000 erythrocytes were examined at a magnification of x 1000 for the presence of micronuclei. The counting of micronuclei in peripheral erythrocytes gave low average incidences, 0.6 per 1000 in the exposed group and 0.1 per 1000 in the control, but statistically significant (P < 0.01) differences were found in the frequency distribution between the control and exposed groups.

  4. Persistent fetal infection with bovine viral diarrhea virus differentially affects maternal blood cell signal transduction pathways.

    PubMed

    Smirnova, Natalia P; Ptitsyn, Andrey A; Austin, Kathleen J; Bielefeldt-Ohmann, Helle; Van Campen, Hana; Han, Hyungchul; van Olphen, Alberto L; Hansen, Thomas R

    2009-02-02

    The consequences of viral infection during pregnancy include impact on fetal and maternal immune responses and on fetal development. Transplacental infection in cattle with noncytopathic bovine viral diarrhea virus (ncpBVDV) during early gestation results in persistently infected (PI) fetuses with life-long viremia and susceptibility to infections. Infection of the fetus during the third trimester or after birth leads to a transient infection cleared by a competent immune system. We hypothesized that ncpBVDV infection and presence of an infected fetus would alter immune response and lead to downregulation of proinflammatory processes in pregnant dams. Naïve pregnant heifers were challenged with ncpBVDV2 on day 75 (PI fetus) and day 175 [transiently infected (TI) fetus] or kept uninfected (healthy control fetus). Maternal blood samples were collected up to day 190 of gestation. Genome-wide microarray analysis of gene expression in maternal peripheral white blood cells, performed on days 160 and 190 of gestation, revealed multiple signal transduction pathways affected by ncpBVDV infection. Acute infection and presence of a TI fetus caused upregulation of the type I interferon (IFN) pathway genes, including dsRNA sensors and IFN-stimulated genes. The presence of a PI fetus caused prolonged downregulation of chemokine receptor 4 (CXCR4) and T cell receptor (TCR) signaling in maternal blood cells. We conclude that: 1) infection with ncpBVDV induces a vigorous type I IFN response, and 2) presence of a PI fetus causes downregulation of important signaling pathways in the blood of the dam, which could have deleterious consequences on fetal development and the immune response.

  5. Quantification of wave reflection using peripheral blood pressure waveforms.

    PubMed

    Kim, Chang-Sei; Fazeli, Nima; McMurtry, M Sean; Finegan, Barry A; Hahn, Jin-Oh

    2015-01-01

    This paper presents a novel minimally invasive method for quantifying blood pressure (BP) wave reflection in the arterial tree. In this method, two peripheral BP waveforms are analyzed to obtain an estimate of central aortic BP waveform, which is used together with a peripheral BP waveform to compute forward and backward pressure waves. These forward and backward waves are then used to quantify the strength of wave reflection in the arterial tree. Two unique strengths of the proposed method are that 1) it replaces highly invasive central aortic BP and flow waveforms required in many existing methods by less invasive peripheral BP waveforms, and 2) it does not require estimation of characteristic impedance. The feasibility of the proposed method was examined in an experimental swine subject under a wide range of physiologic states and in 13 cardiac surgery patients. In the swine subject, the method was comparable to the reference method based on central aortic BP and flow. In cardiac surgery patients, the method was able to estimate forward and backward pressure waves in the absence of any central aortic waveforms: on the average, the root-mean-squared error between actual versus computed forward and backward pressure waves was less than 5 mmHg, and the error between actual versus computed reflection index was less than 0.03.

  6. Kurloff cells in peripheral blood and organs of wild capybaras.

    PubMed

    Jara, Luis Fernando; Sánchez, Jairo Mauricio; Alvarado, Hernán; Nassar-Montoya, Fernando

    2005-04-01

    Peripheral blood and tissue from twenty-two free-ranging, hunter-killed capybaras (Hydrochaeris hydrochaeris) collected between December 1996 and April 1997 in Casanare, Colombia (5 degrees 58'N and 71 degrees 33'W), were examined by light microscopy for Kurloff cells (KCs). Kurloff cells were observed in the blood of one pregnant adult female, and in organs from all the animals, including spleen (21 of 22 animals), liver (18 of 21), lungs (13 of 21), ovary (8 of 11), uterus (7 of 10), bone marrow (13 of 20), kidney (8 of 22), adrenal gland (6 of 20), and lymph node (4 of 14). The anatomic distribution of the KC in the wild capybaras was similar to that of the guinea pig.

  7. Twenty-five years of peripheral blood stem cell transplantation.

    PubMed

    Körbling, Martin; Freireich, Emil J

    2011-06-16

    Peripheral blood stem cell transplantation (PBSCT) is the most common transplantation procedure performed in medicine. Its clinical introduction in 1986 replaced BM as a stem-cell source to approximately 100% in the autologous and to approximately 75% in the allogeneic transplantation setting. This historical overview provides a brief insight into the discovery of circulating hematopoietic stem cells in the early 1960s, the development of apheresis technology, the discovery of hematopoietic growth factors and small molecule CXCR4 antagonist for stem- cell mobilization, and in vivo experimental transplantation studies that eventually led to clinical PBSCT. Also mentioned are the controversies surrounding the engraftment potential of circulating stem cells before acceptance as a clinical modality. Clinical trials comparing the outcome of PBSCT with BM transplantation, registry data analyses, and the role of the National Marrow Donor Program (NMDP) in promoting unrelated blood stem-cell donation are addressed.

  8. Characteristics of peripheral blood monocytes in hereditary xerocytosis and spherocytosis.

    PubMed

    Snyder, L M; Leb, L; Jacobs, J B; Fortier, N L; Reeves, D; Neri, L L; St John, M

    1982-01-01

    Peripheral blood monocytes isolated from patients with congenital hemolytic anemia, hereditary xerocytosis and spherocytosis, demonstrated in vivo engulfment of red cell and platelet fragments. In addition, morphometric studies performed on these monocytes showed an increase in cytoplasmic/nuclear ratio as well as lysosome and phagosome volumes. The production of carbon dioxide from glucose-1-14C in abnormal monocytes was increased (15-80%) but the intracellular values of beta-glucuronidase and esterase activity were similar to control monocytes. Monocyte locomotion assessed in the presence of chemotactic stimuli was found significantly increased (73 +/- 12 monocytes/oil immersion fields vs. 46 +/- 5 for control monocytes). We concluded that the monocytes in hemolytic anemias associated with increased in vitro red cell fragmentation have some features resembling the 'stimulated' monocytes and that this alteration may be due to red blood cell fragment ingestion.

  9. High-resolution ultrasound imaging and noninvasive optoacoustic monitoring of blood variables in peripheral blood vessels

    NASA Astrophysics Data System (ADS)

    Petrov, Irene Y.; Petrov, Yuriy; Prough, Donald S.; Esenaliev, Rinat O.

    2011-03-01

    Ultrasound imaging is being widely used in clinics to obtain diagnostic information non-invasively and in real time. A high-resolution ultrasound imaging platform, Vevo (VisualSonics, Inc.) provides in vivo, real-time images with exceptional resolution (up to 30 microns) using high-frequency transducers (up to 80 MHz). Recently, we built optoacoustic systems for probing radial artery and peripheral veins that can be used for noninvasive monitoring of total hemoglobin concentration, oxyhemoglobin saturation, and concentration of important endogenous and exogenous chromophores (such as ICG). In this work we used the high-resolution ultrasound imaging system Vevo 770 for visualization of the radial artery and peripheral veins and acquired corresponding optoacoustic signals from them using the optoacoustic systems. Analysis of the optoacoustic data with a specially developed algorithm allowed for measurement of blood oxygenation in the blood vessels as well as for continuous, real-time monitoring of arterial and venous blood oxygenation. Our results indicate that: 1) the optoacoustic technique (unlike pure optical approaches and other noninvasive techniques) is capable of accurate peripheral venous oxygenation measurement; and 2) peripheral venous oxygenation is dependent on skin temperature and local hemodynamics. Moreover, we performed for the first time (to the best of our knowledge) a comparative study of optoacoustic arterial oximetry and a standard pulse oximeter in humans and demonstrated superior performance of the optoacoustic arterial oximeter, in particular at low blood flow.

  10. Isolation of foamy viruses from peripheral blood lymphocytes.

    PubMed

    Tobaly-Tapiero, Joëlle; Bittoun, Patricia; Saïb, Ali

    2005-01-01

    The isolation of a retrovirus from peripheral blood lymphocytes/monocytes can be a difficult task, requiring the fulfillment of three essential parameters. First, this viral agent must infect such cells in vivo. Second, these circulating cells should harbor wild-type proviruses. Finally, the viral agent has to express, at least when these cells are cultured in vitro, the structural proteins necessary for the production of viral particles. Foamy viruses (FVs), also known as spumaviruses, are complex retroviruses whose genomic organization has been known since the cloning of the prototypic primate foamy virus type 1. These retroviruses infect most cell lines in culture, but circulating lymphocytes seem to represent their major reservoir in vivo. FV infection leads to the formation of multinucleated giant cells, resulting from the fusion of adjacent infected cells, which present multiple vacuoles giving the monolayer culture a foam aspect. These two features, combined with electron microscopy studies, have helped investigators in their attempt to isolate new FVs. These viruses were described and isolated from different animal species, mostly in nonhuman primates. Here we present the successive steps leading to the isolation of the equine foamy virus from peripheral blood lymphocytes of infected horses.

  11. Mitochondrial DNA copy number in peripheral blood and melanoma risk.

    PubMed

    Shen, Jie; Gopalakrishnan, Vancheswaran; Lee, Jeffrey E; Fang, Shenying; Zhao, Hua

    2015-01-01

    Mitochondrial DNA (mtDNA) copy number in peripheral blood has been suggested as risk modifier in various types of cancer. However, its influence on melanoma risk is unclear. We evaluated the association between mtDNA copy number in peripheral blood and melanoma risk in 500 melanoma cases and 500 healthy controls from an ongoing melanoma study. The mtDNA copy number was measured using real-time polymerase chain reaction. Overall, mean mtDNA copy number was significantly higher in cases than in controls (1.15 vs 0.99, P<0.001). Increased mtDNA copy number was associated with a 1.45-fold increased risk of melanoma (95% confidence interval: 1.12-1.97). Significant joint effects between mtDNA copy number and variables related to pigmentation and history of sunlight exposure were observed. This study supports an association between increased mtDNA copy number and melanoma risk that is independent on the known melanoma risk factors (pigmentation and history of sunlight exposure).

  12. Peripheral blood T and B lymphocytes during acute rheumatic fever.

    PubMed Central

    Lueker, R D; Abdin, Z H; Williams, R C

    1975-01-01

    Proportions and total numbers of thymus-derived (T) and bone marrow-derived (B) peripheral blood lymphocytes were studied in 53 patients with acute rheumatic fever, diagnosed on the basis of modifified Jones criteria. An elevation in both proportions and absolute numbers of cells bearing surface Ig was found in most patients, particularly during the first 7 days after onset. Conversely, T-cell proportions and numbers were often found to be depressed early in the acue phases of rheumatic fever. Proportions of cells bearing surface Ig did not correlate with another B-cell marker, the aggregated gamma globulin receptor, suggesting that such cells bearing surface Ig were not all B lymphocytes. Incuvation for 20 h at 37 per cent C of cells showing high proportions of surface Ig-bearing surface Ig in both normal and rheumatic fever subjects, although there was no appreciable increment in proportions of lymphocytes expressing T-cell markers. Patients with initial attacks showed higher percentages and total numbers of Ig-bearing lymphocytes (P smaller than 0.01) than did those with rneumatic fever recurrences. Elevations in numbers and proportions of peripheral blood lymphocytes bearing Ig appeared to correlate with the relative acute nature of the rheumatic fever attack. PMID:1091658

  13. Genetic analysis in retinoblastoma and peripheral blood correlation.

    PubMed

    Ruiz del Río, N; Abelairas Gómez, J M; Alonso García de la Rosa, F J; Peralta Calvo, J M; de las Heras Martín, A

    2015-12-01

    To determine the importance of intratumoral genetic analysis in the diagnosis of germ-line mutations in patients with retinoblastoma. To underline the importance of performing these genetic tests in every case of retinoblastoma. Intratumoral genetic analysis of RB1 mutation was performed on 17 enucleated eyes that were non-responsive to conservative treatment. Patients had no family history of retinoblastoma, and lesions were always single. The identified mutations were then also studied in peripheral blood analysis. There were 12 (70.6%) cases with positive results in intratumoral analysis. In 8 cases (47.1%) mutation of both RB1 alelli were detected, and in 4 (23.5%) cases only one allele was found mutated. In 5 patients (29.4%) no mutation was identified. In the first hit, mutations comprised 7 frameshift or nonsense and 2 splice, whereas in the second hit, one splice mutation, 2 nonsense and 8 loss of heterozygosity were identified. Among 6 patients where intratumoral analysis detected a single mutation associated with a loss of heterozygosity, the peripheral blood analysis was able to detect the same mutation in 3 cases (50%). Intratumoral genetic analysis of sporadic retinoblastoma can detect germ-line mutations. These patients are at higher risk of bilateralization and development of second tumors or trilateral retinoblastoma. Genetic screening is recommended in every patient diagnosed with retinoblastoma. Copyright © 2013 Sociedad Española de Oftalmología. Published by Elsevier España, S.L.U. All rights reserved.

  14. Peripheral blood CD34+ cell count reliably predicts autograft yield.

    PubMed

    Chapple, P; Prince, H M; Quinn, M; Bertoncello, I; Juneja, S; Wolf, M; Januszewicz, H; Brettell, M; Gardyn, J; Seymour, C; Venter, D

    1998-07-01

    A reliable measure to predict peripheral blood progenitor cell (PBPC) autograft CD34+ cell content is required to optimize the timing of PBPC collection. We prospectively examined the peripheral blood (PB) CD34+ cell count in 59 consecutive patients with various malignancies and analyzed the correlation between the PB CD34+ cell count and various parameters in the PBPC autograft. Two hundred and thirty-five collections were performed with a median of 4.0 collections per patient (range, 2-10). The median PB CD34+ cell count at the time of collection was 39 x 10(6)/1 (range, 0.0-285.6). The PBPC autograft parameters measured were the CD34+ cell, colony-forming unit granulocyte-macrophage (CFU-GM) and mononuclear cell (MNC) content. There was a strong linear correlation between PB CD34+ cells/l and autograft CD34+ cells/kg (r = 0.8477). The correlation with CFU-GM/kg (r = 0.5512) was weaker. There was no correlation between autograft CD34+ cells/kg and PB WBC (r= 0.0684), PB MNC (r = 0.1518) or PB platelet count (r = 0.2010). At our institution we aim to obtain a minimum of 0.5 x 10(6) CD34+ cells/kg with each day of collection. We demonstrate that such a collection can be reliably obtained if the PB CD34+ cell count exceeds 5.0 x 10(6)/l.

  15. Apheresis techniques for collection of peripheral blood progenitor cells.

    PubMed

    Moog, Rainer

    2004-12-01

    The combination of effective mobilisation protocols and efficient use of apheresis machines has caused peripheral blood progenitor cells (PBPC) transplantation to grow rapidly. The development of apheresis technology has improved over the years. Today PBSC procedures have changed towards systems to minimise operator interaction and to reduce the collection of undesired cells such as polymorphonuclear cells and platelets using functionally closed, sterile environments for PBSC collection in keeping with Good Manufacturing Practice guidelines. Blood cell separators with continuous flow technique allow the processing of more blood than intermittent flow devices resulting in higher PBSC yields. Large volume leukapheresis with the processing of 3-4-fold donor's/patient's blood volume can increase the number of collected progenitor cells. Therefore, intermittent flow cell separators are indicated if only single vein access is available. Anticoagulant induced hypocalcaemia is an often observed side effect in long lasting PBPC harvesting and monitoring of electrolytes should be performed especially at the end of the apheresis procedure to supplement low levels of potassium, calcium or magnesium. Refinement and improvement of collection techniques continue to add to the armamentarium of current approaches for cancer and non-malignant conditions and will enable future strategies.

  16. Topical Menthol, Ice, Peripheral Blood Flow, and Perceived Discomfort

    PubMed Central

    Topp, Robert; Ledford, Elizabeth R.; Jacks, Dean E.

    2013-01-01

    Context: Injury management commonly includes decreasing arterial blood flow to the affected site in an attempt to reduce microvascular blood flow and edema and limit the induction of inflammation. Applied separately, ice and menthol gel decrease arterial blood flow, but the combined effects of ice and menthol gel on arterial blood flow are unknown. Objectives: To compare radial artery blood flow, arterial diameter, and perceived discomfort before and after the application of 1 of 4 treatment conditions. Design: Experimental crossover design. Setting: Clinical laboratory. Participants or Other Participants: Ten healthy men, 9 healthy women (mean age = 25.68 years, mean height = 1.73 m, mean weight = 76.73 kg). Intervention(s): Four treatment conditions were randomly applied for 20 minutes to the right forearm of participants on 4 different days separated by at least 24 hours: (1) 3.5 mL menthol gel, (2) 0.5 kg of crushed ice, (3) 3.5 mL of menthol gel and 0.5 kg of crushed ice, or (4) no treatment (control). Main Outcome Measure(s): Using high-resolution ultrasound, we measured right radial artery diameter (cm) and blood flow (mL/min) every 5 minutes for 20 minutes after the treatment was applied. Discomfort with the treatment was documented using a 1-to-10 intensity scale. Results: Radial artery blood flow decreased (P < .05) from baseline in the ice (−20% to −24%), menthol (−17% to −24%), and ice and menthol (−36% to −39%) treatments but not in the control (3% to 9%) at 5, 10, and 15 minutes. At 20 minutes after baseline, only the ice (−27%) and combined ice and menthol (−38%) treatments exhibited reductions in blood flow (P < .05). Discomfort was less with menthol than with the ice treatment at 5, 10, and 20 minutes after application (P < .05). Arterial diameter and heart rate did not change. Conclusions: The application of 3.5 mL of menthol was similar to the application of 0.5 kg of crushed ice in reducing peripheral blood flood. Combining

  17. Topical menthol, ice, peripheral blood flow, and perceived discomfort.

    PubMed

    Topp, Robert; Ledford, Elizabeth R; Jacks, Dean E

    2013-01-01

    Injury management commonly includes decreasing arterial blood flow to the affected site in an attempt to reduce microvascular blood flow and edema and limit the induction of inflammation. Applied separately, ice and menthol gel decrease arterial blood flow, but the combined effects of ice and menthol gel on arterial blood flow are unknown. To compare radial artery blood flow, arterial diameter, and perceived discomfort before and after the application of 1 of 4 treatment conditions. Experimental crossover design. Clinical laboratory. PARTICIPANTS OR OTHER PARTICIPANTS: Ten healthy men, 9 healthy women (mean age = 25.68 years, mean height = 1.73 m, mean weight = 76.73 kg). Four treatment conditions were randomly applied for 20 minutes to the right forearm of participants on 4 different days separated by at least 24 hours: (1) 3.5 mL menthol gel, (2) 0.5 kg of crushed ice, (3) 3.5 mL of menthol gel and 0.5 kg of crushed ice, or (4) no treatment (control). Using high-resolution ultrasound, we measured right radial artery diameter (cm) and blood flow (mL/min) every 5 minutes for 20 minutes after the treatment was applied. Discomfort with the treatment was documented using a 1-to-10 intensity scale. Radial artery blood flow decreased (P < .05) from baseline in the ice (-20% to -24%), menthol (-17% to -24%), and ice and menthol (-36% to -39%) treatments but not in the control (3% to 9%) at 5, 10, and 15 minutes. At 20 minutes after baseline, only the ice (-27%) and combined ice and menthol (-38%) treatments exhibited reductions in blood flow (P < .05). Discomfort was less with menthol than with the ice treatment at 5, 10, and 20 minutes after application (P < .05). Arterial diameter and heart rate did not change. The application of 3.5 mL of menthol was similar to the application of 0.5 kg of crushed ice in reducing peripheral blood flood. Combining crushed ice with menthol appeared to have an additive effect on reducing blood flow.

  18. Mobilization and harvesting of peripheral blood stem cells.

    PubMed

    Moog, Rainer

    2006-05-01

    The use of peripheral blood stem cells (PBSC) as a source of hematopoietic stem cells is steadily increasing and has nearly supplanted bone marrow. The present article reviews mobilization and collection of PBSC as well as its side effects. Specialized harvesting strategies such as large volume leukapheresis (LVL) and pediatric PBSC collection are included in this overview. Under steady state conditions, less than 0.05% of the white blood cells (WBC) are CD34+ cells. Chemotherapy results in a 5-15-fold increase of PBSC. Combining chemotherapy and growth factors increases CD34+ cells up to 6% of WBC. In the allogeneic setting, granulocyte-colony stimulating factor is used alone for PBSC mobilization. Several factors affect the mobilization of PBSC: age, gender, type of growth factor, dose of the growth factor and in the autologous setting, patient's diagnosis, chemotherapy regimen and number of previous chemotherapy cycles or radiation. Harvesting of PBSC can be performed with various blood cell separators using continuous or discontinuous flow technique. Continuous flow separators allow the processing of more blood compared with intermittent flow devices resulting in higher yields of CD34+ cells for transplantation. LVL can be used to increase the CD34+ yield in patients with low CD34+ pre-counts. Processing of more blood in LVL is achieved by an increase of the blood flow rate and an altered anticoagulation regimen. Specialized strategies were developed for pediatric PBSC collection considering the main limiting factors, extracorporeal volume and vascular access. Adverse events in PBSC collection can be subdivided in apheresis associated and mobilization associated side effects. Citrate reactions due to hypocalcemia are frequent during apheresis, especially in pediatric PBSC collection and LVL. Thrombocytopenia is often observed in patients after termination of apheresis due to platelet loss during PBSC harvesting. Muscle and bone pain are frequent adverse events

  19. [Chlamydia trachomaatis DNA in leukocytes of peripheral blood from neonates].

    PubMed

    López-Hurtado, Marcela; Cuevas-Recillas, Karla N; Flores-Salazar, Verónica R; Guerra-Infante, Fernando M

    2015-01-01

    Diagnosis of Chlamydia trachomatis infection in newborns is difficult; however, this diagnosis is performed by cell culture or by detection of IgM antibodies against C. trachomatis. Detection of C. trachomatis DNA in peripheral blood leukocytes using polymer chain reaction (PCR) may be a better tool for the diagnosis of infection by this pathogen. A total of 44 premature newborns, all weighing less than 2500g, were included in the study. A blood sample and nasopharyngeal lavages were obtained from each newborn. Leukocyte DNA was obtained by phenol-chloroform extraction technique. Detection of C. trachomatis was performed by amplifying the ompA gene using the PCR endpoint. Cell culture tests and the detection of IgM antibodies against C. trachomatis by microimmunofluorescence assay were also performed. Twenty newborns were PCR-positive (45.5%), with this test being significantly associated with the presence of pneumonia (RR=2.28; 95%CI: 1.01 to 5.17; P=.035). The cell culture of nasopharyngeal lavage was positive in only 7 samples and no significant association was observed with any clinical or laboratory data. The titer of IgM antibodies against C. trachomatis associated with PCR-positive was 1:32 (RR=2.74; 95%CI: 1.21 to 6.23; P=.008), however this titer was not associated with the presence of pneumonia. DNA detection in peripheral blood leukocytes could be useful for diagnosis of C. trachomatis infection. Copyright © 2014 Elsevier España, S.L.U. y Sociedad Española de Enfermedades Infecciosas y Microbiología Clínica. All rights reserved.

  20. Whole Blood Gene Expression Profiling in Preclinical and Clinical Cattle Infected with Atypical Bovine Spongiform Encephalopathy

    PubMed Central

    Xerxa, Elena; Barbisin, Maura; Chieppa, Maria Novella; Krmac, Helena; Vallino Costassa, Elena; Vatta, Paolo; Simmons, Marion; Caramelli, Maria; Casalone, Cristina; Corona, Cristiano

    2016-01-01

    Prion diseases, such as bovine spongiform encephalopathies (BSE), are transmissible neurodegenerative disorders affecting humans and a wide variety of mammals. Variant Creutzfeldt-Jakob disease (vCJD), a prion disease in humans, has been linked to exposure to BSE prions. This classical BSE (cBSE) is now rapidly disappearing as a result of appropriate measures to control animal feeding. Besides cBSE, two atypical forms (named H- and L-type BSE) have recently been described in Europe, Japan, and North America. Here we describe the first wide-spectrum microarray analysis in whole blood of atypical BSE-infected cattle. Transcriptome changes in infected animals were analyzed prior to and after the onset of clinical signs. The microarray analysis revealed gene expression changes in blood prior to the appearance of the clinical signs and during the progression of the disease. A set of 32 differentially expressed genes was found to be in common between clinical and preclinical stages and showed a very similar expression pattern in the two phases. A 22-gene signature showed an oscillating pattern of expression, being differentially expressed in the preclinical stage and then going back to control levels in the symptomatic phase. One gene, SEL1L3, was downregulated during the progression of the disease. Most of the studies performed up to date utilized various tissues, which are not suitable for a rapid analysis of infected animals and patients. Our findings suggest the intriguing possibility to take advantage of whole blood RNA transcriptional profiling for the preclinical identification of prion infection. Further, this study highlighted several pathways, such as immune response and metabolism that may play an important role in peripheral prion pathogenesis. Finally, the gene expression changes identified in the present study may be further investigated as a fingerprint for monitoring the progression of disease and for developing targeted therapeutic interventions. PMID

  1. Whole Blood Gene Expression Profiling in Preclinical and Clinical Cattle Infected with Atypical Bovine Spongiform Encephalopathy.

    PubMed

    Xerxa, Elena; Barbisin, Maura; Chieppa, Maria Novella; Krmac, Helena; Vallino Costassa, Elena; Vatta, Paolo; Simmons, Marion; Caramelli, Maria; Casalone, Cristina; Corona, Cristiano; Legname, Giuseppe

    2016-01-01

    Prion diseases, such as bovine spongiform encephalopathies (BSE), are transmissible neurodegenerative disorders affecting humans and a wide variety of mammals. Variant Creutzfeldt-Jakob disease (vCJD), a prion disease in humans, has been linked to exposure to BSE prions. This classical BSE (cBSE) is now rapidly disappearing as a result of appropriate measures to control animal feeding. Besides cBSE, two atypical forms (named H- and L-type BSE) have recently been described in Europe, Japan, and North America. Here we describe the first wide-spectrum microarray analysis in whole blood of atypical BSE-infected cattle. Transcriptome changes in infected animals were analyzed prior to and after the onset of clinical signs. The microarray analysis revealed gene expression changes in blood prior to the appearance of the clinical signs and during the progression of the disease. A set of 32 differentially expressed genes was found to be in common between clinical and preclinical stages and showed a very similar expression pattern in the two phases. A 22-gene signature showed an oscillating pattern of expression, being differentially expressed in the preclinical stage and then going back to control levels in the symptomatic phase. One gene, SEL1L3, was downregulated during the progression of the disease. Most of the studies performed up to date utilized various tissues, which are not suitable for a rapid analysis of infected animals and patients. Our findings suggest the intriguing possibility to take advantage of whole blood RNA transcriptional profiling for the preclinical identification of prion infection. Further, this study highlighted several pathways, such as immune response and metabolism that may play an important role in peripheral prion pathogenesis. Finally, the gene expression changes identified in the present study may be further investigated as a fingerprint for monitoring the progression of disease and for developing targeted therapeutic interventions.

  2. Effects of a traditional herbal medicine on peripheral blood flow in women experiencing peripheral coldness: a randomized controlled trial.

    PubMed

    Nishida, Shinji; Eguchi, Eri; Ohira, Tetsuya; Kitamura, Akihiko; Kato, Yukiko Hakariya; Hagihara, Keisuke; Iso, Hiroyasu

    2015-04-02

    In Japan, a traditional herbal medicine, Tokishigyakukagoshuyushokyoto (TJ-38), is often used for the treatment of peripheral coldness, which is a common complaint among Japanese women. However, the effects of this herbal medicine have yet to be examined in a randomized controlled trial. In the current study, the effect of TJ-38 on the peripheral blood flow in women experiencing peripheral coldness was investigated using a parallel-group randomized controlled trial. Fifty-eight women aged 23 to 79 years with peripheral coldness were randomly divided into the intervention or control group. They were examined using cold bathing tests, physical examinations, and questionnaires in January 2010 for the baseline and in March 2010 for the follow-up, and January 2011 and March 2011, respectively. At the baseline, there were no differences in clinical characteristics between the two groups. In the intervention group, peripheral coldness improved after the intervention term; however, it persisted in the control group. Mean values of percentage recovery of the peripheral blood flow after cold bathing tests were 17.2% and -28.2% for the intervention and control groups, respectively (p = 0.007), and the proportions for percentage recovery of >50% were 32% and 0%, respectively (p = 0.0007). Mean values of percent recovery of skin temperature did not differ between the two groups. The present clinical trial supports that a traditional herbal medicine relieves peripheral coldness in women probably through the improvement of peripheral blood flow.

  3. Distribution of cyanide in heart blood, peripheral blood and gastric contents in 21 cyanide related fatalities.

    PubMed

    Rhee, Jongsook; Jung, Jinmi; Yeom, Hyesun; Lee, Hansun; Lee, Sangki; Park, Yoosin; Chung, Heesun

    2011-07-15

    This paper presents 21 cases related to cyanide intoxication by oral ingestion. Cyanide concentrations in biological specimens are especially different from the type of postmortem specimens, and very important in interpreting the cause of death in postmortem forensic toxicology. Besides the detection of cyanide in autopsy specimens, the autopsy findings were unremarkable. Biological samples (0.2mL or equal to less than 10μg of cyanide) were analyzed colorimetrically for cyanide. In a series of 21 cyanide fatalities, the concentration ranges (mean±SD) of cyanide in heart blood, peripheral blood and gastric contents were 0.1-248.6mg/L (38.1±56.6mg/L), 0.3-212.4mg/L (17.1±45.1mg/L) and 2.0-6398.0mg/kg (859.0±1486.2mg/kg), respectively. The ranges of the heart/peripheral blood concentration ratio and gastric contents/peripheral blood concentration ratio were 0.3-10.6 (mean 3.4) and 3.4-402.4 (mean 86.0), respectively. From the difference of cyanide concentration and the concentration ratio of cyanide in different types of postmortem specimens, the possibility of the postmortem redistribution of cyanide and death by oral ingestion of cyanide could be confirmed. We reported cyanide fatal cases along with a review of literature.

  4. A high-quality annotated transcriptome of swine peripheral blood.

    PubMed

    Liu, Haibo; Smith, Timothy P L; Nonneman, Dan J; Dekkers, Jack C M; Tuggle, Christopher K

    2017-06-24

    High throughput gene expression profiling assays of peripheral blood are widely used in biomedicine, as well as in animal genetics and physiology research. Accurate, comprehensive, and precise interpretation of such high throughput assays relies on well-characterized reference genomes and/or transcriptomes. However, neither the reference genome nor the peripheral blood transcriptome of the pig have been sufficiently assembled and annotated to support such profiling assays in this emerging biomedical model organism. We aimed to assemble published and novel RNA-seq data to provide a comprehensive, well-annotated blood transcriptome for pigs by integrating a de novo assembly with a genome-guided assembly. A de novo and a genome-guided transcriptome of porcine whole peripheral blood was assembled with ~162 million pairs of paired-end and ~183 million single-end, trimmed and normalized Illumina RNA-seq reads (~6 billion initial reads from 146 RNA-seq libraries) from five independent studies by using the Trinity and Cufflinks software, respectively. We then removed putative transcripts (PTs) of low confidence from both assemblies and merged the remaining PTs into an integrated transcriptome consisting of 132,928 PTs, with 126,225 (~95%) PTs from the de novo assembly and more than 91% of PTs spliced. In the integrated transcriptome, ~90% and 63% of PTs had significant sequence similarity to sequences in the NCBI NT and NR databases, respectively; 68,754 (~52%) PTs were annotated with 15,965 unique gene ontology (GO) terms; and 7618 PTs annotated with Enzyme Commission codes were assigned to 134 pathways curated by the Kyoto Encyclopedia of Genes and Genomes (KEGG). Full exon-intron junctions of 17,528 PTs were validated by PacBio IsoSeq full-length cDNA reads from 3 other porcine tissues, NCBI pig RefSeq mRNAs and transcripts from Ensembl Sscrofa10.2 annotation. Completeness of the 5' termini of 37,569 PTs was validated by public cap analysis of gene expression (CAGE

  5. [Production of mature red blood cell by using peripheral blood mononuclear cells].

    PubMed

    Jia, Yan-Jun; Liu, Jiang; Zhang, Ke-Ying; Shang, Xiao-Yan; Li, Wei; Wang, Li-Jun; Liu, Na; Wang, Lin; Cui, Shuang; Ni, Lei; Zhao, Bo-Tao; Wang, Dong-Mei; Gao, Song-Ming; Zhang, Zhi-Xin

    2014-10-01

    Most protocols for in vitro producing red blood cells (RBC) use the CD34(+) cells or embryonic stem cells from cord blood, bone marrow or peripheral blood as the start materials. This study was purposed to produce the mature RBC in vitro by using peripheral blood mononuclear cells as start material. The peripheral blood mononuclear cells (PBMNC) were isolated from buffy coat after blood leukapheresis, the mature red blood cells (RBC) were prepared by a 4-step culture protocol. The results showed that after culture by inducing with the different sets of cytokines and supporting by mouse MS-5 cell line, the expansion of PBMNC reached about 1000 folds at the end of the culture. About 90% of cultured RBC were enucleated mature cells which had the comparable morphological characteristics with normal RBC. Colony-forming assays showed that this culture system could stimulate the proliferation of progenitors in PBMNC and differentiate into erythroid cells. The structure and function analysis indicated that the mean cell volume of in vitro cultured RBC was 118 ± 4 fl, which was slight larger than that of normal RBC (80-100 fl); the mean cell hemoglobin was 36 ± 1.2 pg, which was slight higher than that of normal RBC (27-31 pg); the maximal deformation index was 0.46, which approachs level of normal RBC; the glucose-6-phosphate dehydrogenase and pyrurvate kinase levels was consistant with young RBC. It is concluded that PBMNC are feasble, convenient and low-cost source for producing cultured RBC and this culture system is suitable to generate the RBC from PBMNC.

  6. In vitro shear stress-induced platelet activation: sensitivity of human and bovine blood.

    PubMed

    Lu, Qijin; Hofferbert, Bryan V; Koo, Grace; Malinauskas, Richard A

    2013-10-01

    As platelet activation plays a critical role in physiological hemostasis and pathological thrombosis, it is important in the overall hemocompatibility evaluation of new medical devices and biomaterials to assess their effects on platelet function. However, there are currently no widely accepted in vitro test methods to perform this assessment. In an effort to develop effective platelet tests for potential use in medical device evaluation, this study compared the sensitivity of platelet responses to shear stress stimulation of human and bovine blood using multiple platelet activation markers. Fresh whole blood samples anticoagulated with heparin or anticoagulant citrate dextrose, solution A (ACDA) were exposed to shear stresses up to 40 Pa for 2 min using a cone-and-plate rheometer model. Platelet activation was characterized by platelet counts, platelet surface P-selectin expression, and serotonin release into blood plasma. The results indicated that exposure to shear stresses above 20 Pa caused significant changes in all three of the platelet markers for human blood and that the changes were usually greater with ACDA anticoagulation than with heparin. In contrast, for bovine blood, the markers did not change with shear stress stimulation except for plasma serotonin in heparin anticoagulated blood. The differences observed between human and bovine platelet responses suggest that the value of using bovine blood for in vitro platelet testing to evaluate devices may be limited.

  7. Angiogenic conditioning of peripheral blood mononuclear cells promotes fracture healing

    PubMed Central

    Mifuji, K.; Kamei, N.; Tanaka, R.; Arita, K.; Mizuno, H.; Asahara, T.; Adachi, N.; Ochi, M.

    2017-01-01

    Objectives The objective of this study was to investigate the therapeutic effect of peripheral blood mononuclear cells (PBMNCs) treated with quality and quantity control culture (QQ-culture) to expand and fortify angiogenic cells on the acceleration of fracture healing. Methods Human PBMNCs were cultured for seven days with the QQ-culture method using a serum-free medium containing five specific cytokines and growth factors. The QQ-cultured PBMNCs (QQMNCs) obtained were counted and characterised by flow cytometry and real-time polymerase chain reaction (RT-PCR). Angiogenic and osteo-inductive potentials were evaluated using tube formation assays and co-culture with mesenchymal stem cells with osteo-inductive medium in vitro. In order to evaluate the therapeutic potential of QQMNCs, cells were transplanted into an immunodeficient rat femur nonunion model. The rats were randomised into three groups: control; PBMNCs; and QQMNCs. The fracture healing was evaluated radiographically and histologically. Results The total number of PBMNCs was decreased after QQ-culture, however, the number of CD34+ and CD206+ cells were found to have increased as assessed by flow cytometry analysis. In addition, gene expression of angiogenic factors was upregulated in QQMNCs. In the animal model, the rate of bone union was higher in the QQMNC group than in the other groups. Radiographic scores and bone volume were significantly associated with the enhancement of angiogenesis in the QQMNC group. Conclusion We have demonstrated that QQMNCs have superior potential to accelerate fracture healing compared with PBMNCs. The QQMNCs could be a promising option for fracture nonunion. Cite this article: K. Mifuji, M. Ishikawa, N. Kamei, R. Tanaka, K. Arita, H. Mizuno, T. Asahara, N. Adachi, M. Ochi. Angiogenic conditioning of peripheral blood mononuclear cells promotes fracture healing. Bone Joint Res 2017;6: 489–498. DOI: 10.1302/2046-3758.68.BJR-2016-0338.R1. PMID:28835445

  8. Peripheral organ perfusion augmentation during left ventricular failure. A controlled bovine comparison between the intraaortic balloon pump and the Hemopump.

    PubMed Central

    Baldwin, R T; Radovancević, B; Conger, J L; Matsuwaka, R; Duncan, J M; Vaughn, W K; Wampler, R K; Frazier, O H

    1993-01-01

    Despite the use of inotropic therapy and the intraaortic balloon pump (IABP), inadequate peripheral organ perfusion and subsequent multiorgan failure from left ventricular dysfunction is a major cause of death following cardiac surgery. To compare the end-organ perfusion provided by the IABP with that of the recently developed Hemopump Cardiac Assist System, blood flow from visceral organs was measured by ultrasonic flow probes during separate periods of support with each of these pumps. Ten calves underwent coronary artery ligations with beta-receptor blockade; hemodynamic parameters were recorded before the induction of failure, during unsupported cardiac failure, and during Hemopump and IABP support. Improvement in mean cardiac output, mixed venous oxygen saturation, and pulmonary artery wedge pressure was significantly greater (p < 0.05) during Hemopump support than during IABP support. Renal artery flow was significantly greater during Hemopump support (276 +/- 74.2 cc/min) than during IABP support (164 +/- 79.6 cc/min). Hepatic artery flow was significantly greater during Hemopump support (34.7 +/- 25.7 cc/min) than during IABP support (24.4 +/- 18.9 cc/min), and portal vein flow was significantly greater during Hemopump support (1588 +/- 315 cc/min) than IABP support (1259 +/- 310 cc/min). There were no significant differences, however, between carotid artery flow during Hemopump support (292 +/- 171 cc/min) and that during IABP support (317 +/- 204 cc/min). We conclude that renal, hepatic, and mesenteric perfusion provided by the nonpulsatile Hemopump is superior to that of the IABP in this bovine model of left ventricular failure. Therefore, the Hemopump may be more effective in preventing multiorgan failure during recovery of ventricular function. PMID:8298324

  9. Automatic recognition of five types of white blood cells in peripheral blood.

    PubMed

    Rezatofighi, Seyed Hamid; Soltanian-Zadeh, Hamid

    2011-06-01

    This paper proposes image processing algorithms to recognize five types of white blood cells in peripheral blood automatically. First, a method based on Gram-Schmidt orthogonalization is proposed along with a snake algorithm to segment nucleus and cytoplasm of the cells. Then, a variety of features are extracted from the segmented regions. Next, most discriminative features are selected using a Sequential Forward Selection (SFS) algorithm and performances of two classifiers, Artificial Neural Network (ANN) and Support Vector Machine (SVM), are compared. The results demonstrate that the proposed methods are accurate and sufficiently fast to be used in hematological laboratories.

  10. Peripheral Blood Transcriptomic Signatures of Fasting Glucose and Insulin Concentrations.

    PubMed

    Chen, Brian H; Hivert, Marie-France; Peters, Marjolein J; Pilling, Luke C; Hogan, John D; Pham, Lisa M; Harries, Lorna W; Fox, Caroline S; Bandinelli, Stefania; Dehghan, Abbas; Hernandez, Dena G; Hofman, Albert; Hong, Jaeyoung; Joehanes, Roby; Johnson, Andrew D; Munson, Peter J; Rybin, Denis V; Singleton, Andrew B; Uitterlinden, André G; Ying, Saixia; Melzer, David; Levy, Daniel; van Meurs, Joyce B J; Ferrucci, Luigi; Florez, Jose C; Dupuis, Josée; Meigs, James B; Kolaczyk, Eric D

    2016-12-01

    Genome-wide association studies (GWAS) have successfully identified genetic loci associated with glycemic traits. However, characterizing the functional significance of these loci has proven challenging. We sought to gain insights into the regulation of fasting insulin and fasting glucose through the use of gene expression microarray data from peripheral blood samples of participants without diabetes in the Framingham Heart Study (FHS) (n = 5,056), the Rotterdam Study (RS) (n = 723), and the InCHIANTI Study (Invecchiare in Chianti) (n = 595). Using a false discovery rate q <0.05, we identified three transcripts associated with fasting glucose and 433 transcripts associated with fasting insulin levels after adjusting for age, sex, technical covariates, and complete blood cell counts. Among the findings, circulating IGF2BP2 transcript levels were positively associated with fasting insulin in both the FHS and RS. Using 1000 Genomes-imputed genotype data, we identified 47,587 cis-expression quantitative trait loci (eQTL) and 6,695 trans-eQTL associated with the 433 significant insulin-associated transcripts. Of note, we identified a trans-eQTL (rs592423), where the A allele was associated with higher IGF2BP2 levels and with fasting insulin in an independent genetic meta-analysis comprised of 50,823 individuals. We conclude that integration of genomic and transcriptomic data implicate circulating IGF2BP2 mRNA levels associated with glucose and insulin homeostasis. © 2016 by the American Diabetes Association.

  11. In-Depth Profiling of the Peripheral Blood Mononuclear Cells Proteome for Clinical Blood Proteomics

    PubMed Central

    Končarević, Saša; Lößner, Christopher; Pike, Ian; Zucht, Hans-Dieter

    2014-01-01

    Peripheral blood mononuclear cells (PBMCs) are an easy accessible cellular part of the blood organ and, along with platelets, represent the only site of active gene expression in blood. These cells undergo immunophenotypic changes in various diseases and represent a peripheral source of monitoring gene expression and posttranslational modifications relevant to many diseases. Little is known about the source of many blood proteins and we hypothesise that release from PBMCs through active and passive mechanisms may account for a substantial part of the plasma proteome. The use of state-of-the-art proteomic profiling methods in PBMCs will enable minimally invasive monitoring of disease progression or response to treatment and discovery of biomarkers. To achieve this goal, detailed mapping of the PBMC proteome using a sensitive, robust, and quantitative methodological setup is required. We have applied an indepth gel-free proteomics approach using tandem mass tags (TMT), unfractionated and SCX fractionated PBMC samples, and LC-MS/MS with various modulations. This study represents a benchmark in deciphering the PBMC proteome as we provide a deep insight by identifying 4129 proteins and 25503 peptides. The identified proteome defines the scope that enables PBMCs to be characterised as cellular major biomarker pool within the blood organ. PMID:24724028

  12. In-depth profiling of the peripheral blood mononuclear cells proteome for clinical blood proteomics.

    PubMed

    Končarević, Saša; Lößner, Christopher; Kuhn, Karsten; Prinz, Thorsten; Pike, Ian; Zucht, Hans-Dieter

    2014-01-01

    Peripheral blood mononuclear cells (PBMCs) are an easy accessible cellular part of the blood organ and, along with platelets, represent the only site of active gene expression in blood. These cells undergo immunophenotypic changes in various diseases and represent a peripheral source of monitoring gene expression and posttranslational modifications relevant to many diseases. Little is known about the source of many blood proteins and we hypothesise that release from PBMCs through active and passive mechanisms may account for a substantial part of the plasma proteome. The use of state-of-the-art proteomic profiling methods in PBMCs will enable minimally invasive monitoring of disease progression or response to treatment and discovery of biomarkers. To achieve this goal, detailed mapping of the PBMC proteome using a sensitive, robust, and quantitative methodological setup is required. We have applied an indepth gel-free proteomics approach using tandem mass tags (TMT), unfractionated and SCX fractionated PBMC samples, and LC-MS/MS with various modulations. This study represents a benchmark in deciphering the PBMC proteome as we provide a deep insight by identifying 4129 proteins and 25503 peptides. The identified proteome defines the scope that enables PBMCs to be characterised as cellular major biomarker pool within the blood organ.

  13. Effect of Peripheral Edema on Oscillometric Blood Pressure Measurement

    PubMed Central

    Ghaffari, Shamsi; Malaki, Majid; Rezaeifar, Afshin; Abdollahi Fakhim, Shahin

    2014-01-01

    Introduction: Blood pressure (BP) measurement is essential for epidemiological studies and clinical decisions. It seems that tissue characteristics can affect BP results and we try to find edema effect on BP results taken by different methods. Methods: BP of 55 children before open heart surgery were measured and compared according to three methods: Arterial as standard and reference, oscillometric and auscultatory methods. Peripheral edema as a tissue characteristic was defined in higher than +2 as marked edema and in equal or lower than +2 as no edema. Statistical analyses: data was expressed as Mean and 95% of confidence interval (CI 95%). Comparison of two groups was performed by T independent test and of more than two groups by ANOVA test. Mann–Whitney U and paired T-test were used for serially comparisons of changes. P less than 0.05 was considered significant. Results: Fifty five children aged 29.4±3.9 months were divided into two groups: 10 children with peripheral edema beyond +2 and 45 cases without edema. Oscillometric method overestimated systolic BP and the Mean (CI 95%) difference of oscillometric to arterial was 4.8 (8/-1, P=0.02) in edematous and 4.2 (7/1, p=0.004) in non edematous. Oscillometric method underestimated diastolic BP as -9 (-1.8/-16.5, P=0.03) in edematous group and 2.6 (-0.7/+5, P= 0.2) in non edematous compared to arterial method. Conclusion: Oscillometric device standards cannot cover all specific clinical conditions. It underestimates diastolic BP significantly in edematous children, which was 9.2 mmHg in average beyond the acceptable standards. PMID:25610552

  14. Sialic acid, ferritin and CEA levels in peripheral blood and blood draining from the tumor in breast cancer.

    PubMed

    Monti, M; Catania, S; Locatelli, E; Scazzoso, A; Calzaferri, G; Cunietti, E

    1988-01-01

    Concentrations of total serum N-acetyl-neuraminic acid, carcinoembryonic antigen, ferritin, lactate dehydrogenase, creatine phosphokinase and total proteins were measured in both tumor drainage blood (axillary vein) and in peripheral blood taken during surgery from 44 breast cancer patients. There were no significant differences in any of the markers between mean values in peripheral and tumor drainage blood, between cancer patients and healthy controls, between patients with or without axillary lymph node metastases, or according to the site of breast mass.

  15. Gene expression profiling of peripheral blood mononuclear cells in the setting of peripheral arterial disease

    PubMed Central

    2012-01-01

    Background Peripheral arterial disease (PAD) is a relatively common manifestation of systemic atherosclerosis that leads to progressive narrowing of the lumen of leg arteries. Circulating monocytes are in contact with the arterial wall and can serve as reporters of vascular pathology in the setting of PAD. We performed gene expression analysis of peripheral blood mononuclear cells (PBMC) in patients with PAD and controls without PAD to identify differentially regulated genes. Methods PAD was defined as an ankle brachial index (ABI) ≤0.9 (n = 19) while age and gender matched controls had an ABI > 1.0 (n = 18). Microarray analysis was performed using Affymetrix HG-U133 plus 2.0 gene chips and analyzed using GeneSpring GX 11.0. Gene expression data was normalized using Robust Multichip Analysis (RMA) normalization method, differential expression was defined as a fold change ≥1.5, followed by unpaired Mann-Whitney test (P < 0.05) and correction for multiple testing by Benjamini and Hochberg False Discovery Rate. Meta-analysis of differentially expressed genes was performed using an integrated bioinformatics pipeline with tools for enrichment analysis using Gene Ontology (GO) terms, pathway analysis using Kyoto Encyclopedia of Genes and Genomes (KEGG), molecular event enrichment using Reactome annotations and network analysis using Ingenuity Pathway Analysis suite. Extensive biocuration was also performed to understand the functional context of genes. Results We identified 87 genes differentially expressed in the setting of PAD; 40 genes were upregulated and 47 genes were downregulated. We employed an integrated bioinformatics pipeline coupled with literature curation to characterize the functional coherence of differentially regulated genes. Conclusion Notably, upregulated genes mediate immune response, inflammation, apoptosis, stress response, phosphorylation, hemostasis, platelet activation and platelet aggregation. Downregulated genes included several genes from

  16. Peripheral blood methylation profiling of female Crohn's disease patients.

    PubMed

    Li Yim, Andrew Y F; Duijvis, Nicolette W; Zhao, Jing; de Jonge, Wouter J; D'Haens, Geert R A M; Mannens, Marcel M A M; Mul, Adri N P M; Te Velde, Anje A; Henneman, Peter

    2016-01-01

    Crohn's disease (CD) is a chronic inflammatory disorder belonging to the inflammatory bowel diseases (IBD). CD affects distinct parts of the gastrointestinal tract, leading to symptoms including diarrhea, fever, abdominal pain, weight loss, and anemia. The aim of this study was to assess whether the DNA methylome of peripheral blood cells can be associated with CD in women. Samples were obtained from 18 female patients with histologically confirmed ileal or ileocolic CD and 25 healthy age- and gender-matched controls (mean age and standard deviation: 30.5 ± 6.5 years for both groups). Genome-wide DNA methylation was determined using the Illumina HumanMethylation 450k BeadChip. Our analysis implicated 4287 differentially methylated positions (DMPs; corrected p < 0.05) that are associated to 2715 unique genes. Gene ontology enrichment analysis revealed significant enrichment of our DMPs in immune response processes and inflammatory pathways. Of the 4287 DMPs, 32 DMPs were located on chromosome X with several hits for MIR223 and PABPC5. Comparison with previously performed (epi)genome-wide studies revealed that we replicated 33 IBD-associated genes. In addition to DMPs, we found eight differentially methylated regions (DMRs). CD patients display a characteristic DNA methylation landscape, with the differentially methylated genes being implicated in immune response. Additionally, DMPs were found on chromosome X suggesting X-linked manifestations of CD that could be associated with female-specific symptoms.

  17. Constitutive apoptosis in equine peripheral blood neutrophils in vitro

    PubMed Central

    Brazil, Timothy J.; Dixon, Padraic M.; Haslett, Christopher; Murray, Joanna; McGorum, Bruce C.

    2014-01-01

    The aim of this study was to characterise constitutive apoptosis in equine peripheral blood neutrophils, including assessment of factors that potentially modulate neutrophil survival through alteration of the rate of constitutive apoptosis. Cells underwent spontaneous time-dependent constitutive apoptosis when aged in culture for up to 36 h, developing the structural and functional features of apoptosis observed in many cell types, including human neutrophils. Neutrophils undergoing apoptosis also had diminished zymosan activated serum (ZAS)-stimulated chemiluminescence, but maintained responsiveness to phorbol myristate acetate (PMA). The constitutive rate of equine neutrophil apoptosis was promoted by lipopolysaccharide (LPS), tumour necrosis factor α and phagocytosis of opsonised ovine erythrocytes, while it was inhibited by dexamethasone and ZAS (a source of C5a). Formyl-Met-Leu-Phe, leukotriene B4, platelet activating factor and PMA had no demonstrable effect on equine neutrophil apoptosis. There was a difference between equine and human neutrophil apoptosis in response to LPS and the time-dependence of the response to dexamethasone. PMID:25239298

  18. Constitutive apoptosis in equine peripheral blood neutrophils in vitro.

    PubMed

    Brazil, Timothy J; Dixon, Padraic M; Haslett, Christopher; Murray, Joanna; McGorum, Bruce C

    2014-12-01

    The aim of this study was to characterise constitutive apoptosis in equine peripheral blood neutrophils, including assessment of factors that potentially modulate neutrophil survival through alteration of the rate of constitutive apoptosis. Cells underwent spontaneous time-dependent constitutive apoptosis when aged in culture for up to 36 h, developing the structural and functional features of apoptosis observed in many cell types, including human neutrophils. Neutrophils undergoing apoptosis also had diminished zymosan activated serum (ZAS)-stimulated chemiluminescence, but maintained responsiveness to phorbol myristate acetate (PMA). The constitutive rate of equine neutrophil apoptosis was promoted by lipopolysaccharide (LPS), tumour necrosis factor α and phagocytosis of opsonised ovine erythrocytes, while it was inhibited by dexamethasone and ZAS (a source of C5a). Formyl-Met-Leu-Phe, leukotriene B4, platelet activating factor and PMA had no demonstrable effect on equine neutrophil apoptosis. There was a difference between equine and human neutrophil apoptosis in response to LPS and the time-dependence of the response to dexamethasone.

  19. [Collection and transfusion of peripheral blood stem cells].

    PubMed

    Höcker, P; Wagner, A

    1991-01-01

    Harvesting of peripheral blood stem cells (PBSC) by cytapheresis was performed in 57 patients, who underwent chemotherapy. The best yields were obtained when the leukocyte count was above 1 x 10(9)/l and the platelet count was raised above 80 x 10(9)/l. Using a Haemonetics V-50 or a Baxter CS-3000, 374 PBSC-aphereses were performed with a median of six aphereses per patient. The median number of PBSC (CFU-GM) retransfused in 22 patients who received PBSC for hematological reconstitution only was 3.26 x 10(4)/kg. For 22 patients who received autologous bone marrow plus BPSC, the median number of retransfused PBSC was 2.14 x 10(4)/kg. Myeloid engraftment was achieved in all patients, but megakaryopoiesis was delayed when the number of PBSC was less than 5.0 x 10(4)/kg. The results demonstrate that harvesting of a sufficient number of PBSC after chemotherapy is feasible but further measures like the use of rh GM-CSF will be necessary to reduce apheresis procedures and to obtain high yields to ensure rapid and complete engraftment.

  20. Sex differences in the human peripheral blood transcriptome

    PubMed Central

    2014-01-01

    Background Genomes of men and women differ in only a limited number of genes located on the sex chromosomes, whereas the transcriptome is far more sex-specific. Identification of sex-biased gene expression will contribute to understanding the molecular basis of sex-differences in complex traits and common diseases. Results Sex differences in the human peripheral blood transcriptome were characterized using microarrays in 5,241 subjects, accounting for menopause status and hormonal contraceptive use. Sex-specific expression was observed for 582 autosomal genes, of which 57.7% was upregulated in women (female-biased genes). Female-biased genes were enriched for several immune system GO categories, genes linked to rheumatoid arthritis (16%) and genes regulated by estrogen (18%). Male-biased genes were enriched for genes linked to renal cancer (9%). Sex-differences in gene expression were smaller in postmenopausal women, larger in women using hormonal contraceptives and not caused by sex-specific eQTLs, confirming the role of estrogen in regulating sex-biased genes. Conclusions This study indicates that sex-bias in gene expression is extensive and may underlie sex-differences in the prevalence of common diseases. PMID:24438232

  1. Culture and characterisation of equine peripheral blood mesenchymal stromal cells.

    PubMed

    Spaas, Jan H; De Schauwer, Catharina; Cornillie, Pieter; Meyer, Evelyne; Van Soom, Ann; Van de Walle, Gerlinde R

    2013-01-01

    Although the use of mesenchymal stromal cells (MSCs) for the treatment of orthopaedic injuries in horses has been reported, no official guidelines exist that classify a particular cell as an equine MSC. Given the limited characterisation of peripheral blood (PB)-derived equine MSCs in particular, this study aimed to provide more detailed information in relation to this cell type. Mesenchymal stromal cells were isolated from equine PB samples and colony forming unit (CFU) assays as well as population doubling times (PDTs) (from P(0) to P(10)) were performed. Two types of colonies, 'fingerprint' and dispersed, could be observed based on macroscopic and microscopic features. Moreover, after an initial lag phase (as indicated by a negative PDT at P(0) to P(1)) the MSCs divided rapidly as indicated by a positive PDT at all further passages. Immunophenotyping was carried out with trypsin- as well as with accutase-detached MSC to evaluate potential trypsin-sensitive epitope destruction on particular antigens. Isolated MSC were positive for CD29, CD44, CD90 and CD105, and negative for CD45, CD79α, MHC II and a monocyte/macrophage marker, irrespective of the cell detaching agent used. Trilineage differentiation of the MSCs towards osteoblasts, chondroblasts and adipocytes was confirmed using a range of histochemical stains. Copyright © 2012 Elsevier Ltd. All rights reserved.

  2. Peripheral blood eosinophil counts predict the prognosis of drug eruptions.

    PubMed

    Yang, J; Yang, X; Li, M

    2013-01-01

    Previous studies indicated that eosinophils infiltrate the skin during drug eruptions and that counts may become elevated in circulation. However, little is known about the role of eosinophils in the prognosis of patients with drug eruption. This study aims to investigate the correlation between circulating eosinophil counts and the prognosis of patients with drug eruption. A total of 113 patients were enrolled in this study. Clinical features, peripheral blood eosinophil counts, and liver function were analyzed in patients and controls. Our study indicated that eosinophils changed dynamically in different types of drug eruption and that mean eosinophil counts in patients with erythema multiforme-type drug eruption were significantly higher than in patients with other types of eruption. Most patients with eosinophilia had poor liver function, prolonged corticosteroid use, and extended hospitalization, all of which indicate severe disease. Our data suggest that circulating eosinophil counts were positively correlated with the severity of the drug eruption. Therefore, corticosteroids may be needed to treat patients with eosinophilia in clinical practice.

  3. Fish peripheral blood mononuclear cells preparation for future monitoring applications.

    PubMed

    Pierrard, Marie-Aline; Roland, Kathleen; Kestemont, Patrick; Dieu, Marc; Raes, Martine; Silvestre, Frédéric

    2012-07-15

    Fish species possess many specific characteristics that support their use in ecotoxicology. Widely used in clinical research, peripheral blood mononuclear cells (PBMCs) can reasonably be exploited as relevant target cells in the assessment of environmental chemical toxicity. The current article focuses on the methods necessary to isolate, characterize, and culture fish PBMCs. These procedures were successfully applied on an endangered species, the European eel (Anguilla anguilla L.), and on an economically important and worldwide exported species, the Asian catfish (Pangasianodon hypophthalmus S.). Proteomic approaches can be useful to screen xenobiotic exposure at the protein expression level, giving the opportunity to develop early warning signals thanks to molecular signatures of toxicity. To date, a major limitation of proteomic analyses is that most protein expression profiles often reveal the same predominant and frequently differentially expressed families of proteins regardless of the experimental stressing conditions. The current study describes a methodology to get a postnuclear fraction of high quality isolated from fish PBMCs in order to perform subsequent subproteomic analyses. Applied on samples from eel, the subproteomic analysis (two-dimensional differential in-gel electrophoresis) allowed the identification by liquid chromatography-tandem mass spectrometry and searches in the full NCBInr (National Center for Biotechnology Information nonredundant) database of 66 proteins representing 36 different proteins validated through Peptide and Protein Prophet of Scaffold software.

  4. Evaluation of endotoxin (LPS) activity in bovine blood using neutrophil dependent chemiluminescence

    USDA-ARS?s Scientific Manuscript database

    The purpose of this study was to evaluate the applicability of a neutrophil chemiluminescence-based assay for the measurement of LPS stimulatory activity in bovine whole blood. The assay is based on the capacity for LPS to trigger the respiratory oxidative burst activity (RBA) of autologous neutroph...

  5. Mucocutaneous blood contact: blood release behavior of safety peripheral intravenous catheters.

    PubMed

    Wittmann, Andreas; Köver, Jan; Kralj, Nenad; Gasthaus, Klaus; Tosch, Marco; Hofmann, Friedrich

    2013-12-01

    Protection against needlestick injuries has significantly improved in recent years thanks to so-called "safety devices." However, a potential drawback occasionally reported by users is a risk of blood splashing. If this blood comes in contact with the mucous membranes, it could lead to an infection. Five safety peripheral intravenous catheter brands were examined in a laboratory test. To simulate the extreme situations, which may arise through human use, the introducer needle was withdrawn from the catheter at 2 different angles whereby an industrial robot was used to simulate the sequence of this movement. Each brand was tested 30 times. The experiment was carried out using radioactively labeled human whole blood. The measurements for the transmitted volume of blood was taken both from an artificial head and from a surface measuring 18.5 cm by 26.5 cm at a height of 30 cm above the catheter; scintigraphy was used to take the measurements. The volume of blood droplets potentially splashing into the mucous membranes was in the range of 1 nL. For normal virus concentrations in the blood of sick patients, this dose is too small to cause hepatitis C and HIV. Copyright © 2013 Association for Professionals in Infection Control and Epidemiology, Inc. Published by Mosby, Inc. All rights reserved.

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

    PubMed

    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.

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

    PubMed Central

    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

  8. Red Blood Cell Supernatant Potentiates LPS-Induced Proinflammatory Cytokine Response From Peripheral Blood Mononuclear Cells

    PubMed Central

    Nydam, Trevor L.; Clarke, Jason H.; Banerjee, Anirban; Silliman, Christopher C.; McCarter, Martin D.

    2009-01-01

    Allogeneic blood transfusion has an immunomodulatory capacity on its recipients through accumulation of immunologically active substances with blood storage, and prestorage leukoreduction reduces many of these mediators. We investigated lipopolysaccharide (LPS)-induced cytokine response of peripheral blood mononuclear cells (PBMCs) exposed to packed red blood cell (PRBC) supernatants from leukoreduced (LR) or non-leukoreduced (NLR) units with variable duration of storage. PRBC units were collected with or without leukoreduction on Day 0 before routine storage. The plasma fraction (supernatant) was isolated from LR and NLR units after 1 day (D1) or 42 days (D42) of storage and exposed to PBMCs versus control media for 24 h, then with LPS for an additional 24 h. Cell supernatants were analyzed for IL-1β, IL-6, IL-8, IL-10, and TNF-α by cytokine bead array. IL-1β, TNF-α, and IL-6 were significantly elevated in PRBC groups versus control. D42 NLR PRBC supernatant significantly increased secretion of IL-1β and IL-6 compared to D1 NLR PRBC supernatant. LR significantly attenuated the cytokine response of IL-1β. Thus, PRBC supernatant potentiates proinflammatory LPS-induced cytokine secretion from PBMCs. This response is accentuated with storage duration and partially attenuated with leukoreduction. These findings may partially explain the immune activation seen clinically after blood transfusion. PMID:19441884

  9. The Transcriptome of Equine Peripheral Blood Mononuclear Cells

    PubMed Central

    Pacholewska, Alicja; Drögemüller, Michaela; Klukowska-Rötzler, Jolanta; Lanz, Simone; Hamza, Eman; Dermitzakis, Emmanouil T.; Marti, Eliane; Gerber, Vincent

    2015-01-01

    Complete transcriptomic data at high resolution are available only for a few model organisms with medical importance. The gene structures of non-model organisms are mostly computationally predicted based on comparative genomics with other species. As a result, more than half of the horse gene models are known only by projection. Experimental data supporting these gene models are scarce. Moreover, most of the annotated equine genes are single-transcript genes. Utilizing RNA sequencing (RNA-seq) the experimental validation of predicted transcriptomes has become accessible at reasonable costs. To improve the horse genome annotation we performed RNA-seq on 561 samples of peripheral blood mononuclear cells (PBMCs) derived from 85 Warmblood horses. The mapped sequencing reads were used to build a new transcriptome assembly. The new assembly revealed many alternative isoforms associated to known genes or to those predicted by the Ensembl and/or Gnomon pipelines. We also identified 7,531 transcripts not associated with any horse gene annotated in public databases. Of these, 3,280 transcripts did not have a homologous match to any sequence deposited in the NCBI EST database suggesting horse specificity. The unknown transcripts were categorized as coding and noncoding based on predicted coding potential scores. Among them 230 transcripts had high coding potential score, at least 2 exons, and an open reading frame of at least 300 nt. We experimentally validated 9 new equine coding transcripts using RT-PCR and Sanger sequencing. Our results provide valuable detailed information on many transcripts yet to be annotated in the horse genome. PMID:25790166

  10. Osteoclastogenic Potential of Peripheral Blood Mononuclear Cells in Cleidocranial Dysplasia

    PubMed Central

    Faienza, Maria Felicia; Ventura, Annamaria; Piacente, Laura; Ciccarelli, Maria; Gigante, Margherita; Gesualdo, Loreto; Colucci, Silvia; Cavallo, Luciano; Grano, Maria; Brunetti, Giacomina

    2014-01-01

    Cleidocranial dysplasia (CCD) is an autosomal dominant skeletal dysplasia characterized by hypoplastic or aplastic clavicles, dental abnormalities, and delayed closure of the cranial sutures. In addition, mid-face hypoplasia, short stature, skeletal anomalies and osteoporosis are common. We aimed to evaluate osteoclastogenesis in a child (4 years old), who presented with clinical signs of CCD and who have been diagnosed as affected by deletion of RUNX2, master gene in osteoblast differentiation, but also affecting T cell development and indirectly osteoclastogenesis. The results of this study may help to understand whether in this disease is present an alteration in the bone-resorptive cells, the osteoclasts (OCs). Unfractionated and T cell-depleted Peripheral Blood Mononuclear Cells (PBMCs) from patient were cultured in presence/absence of recombinant human M-CSF and RANKL. At the end of the culture period, OCs only developed following the addition of M-CSF and RANKL. Moreover, real-time PCR experiment showed that freshly isolated T cells expressed the osteoclastogenic cytokines (RANKL and TNFα) at very low level, as in controls. This is in accordance with results arising from flow cytometry experiments demonstrating an high percentage of circulating CD4+CD28+ and CD4+CD27+ T cells, not able to produce osteoclastogenic cytokines. Also RANKL, OPG and CTX serum levels in CCD patient are similar to controls, whereas QUS measurements showed an osteoporotic status (BTT-Z score -3.09) in the patient. In conclusions, our findings suggest that the heterozygous deletion of RUNX2 in this CCD patient did not alter the osteoclastogenic potential of PBMCs in vitro. PMID:24578613

  11. Secretome of Peripheral Blood Mononuclear Cells Enhances Wound Healing

    PubMed Central

    Haider, Thomas; Gschwandtner, Maria; Werba, Gregor; Barresi, Caterina; Zimmermann, Matthias; Golabi, Bahar; Tschachler, Erwin; Ankersmit, Hendrik Jan

    2013-01-01

    Non-healing skin ulcers are often resistant to most common therapies. Treatment with growth factors has been demonstrated to improve closure of chronic wounds. Here we investigate whether lyophilized culture supernatant of freshly isolated peripheral blood mononuclear cells (PBMC) is able to enhance wound healing. PBMC from healthy human individuals were prepared and cultured for 24 hours. Supernatants were collected, dialyzed and lyophilized (SECPBMC). Six mm punch biopsy wounds were set on the backs of C57BL/6J-mice and SECPBMC containing emulsion or controls were applied daily for three days. Morphology and neo-angiogenesis were analyzed by H&E-staining and CD31 immuno-staining, respectively. In vitro effects on diverse skin cells were investigated by migration assays, cell cycle analysis, and tube formation assay. Signaling pathways were analyzed by Western blot analysis. Application of SECPBMC on 6 mm punch biopsy wounds significantly enhanced wound closure. H&E staining of the wounds after 6 days revealed that wound healing was more advanced after application of SECPBMC containing emulsion. Furthermore, there was a massive increase in CD31 positive cells, indicating enhanced neo-angiogenesis. In primary human fibroblasts (FB) and keratinocytes (KC) migration but not proliferation was induced. In endothelial cells (EC) SECPBMC induced proliferation and tube-formation in a matrigel-assay. In addition, SECPBMC treatment of skin cells led to the induction of multiple signaling pathways involved in cell migration, proliferation and survival. In summary, we could show that emulsions containing the secretome of PBMC derived from healthy individuals accelerates wound healing in a mouse model and induce wound healing associated mechanisms in human primary skin cells. The formulation and use of such emulsions might therefore represent a possible novel option for the treatment of non-healing skin ulcers. PMID:23533667

  12. Secretome of peripheral blood mononuclear cells enhances wound healing.

    PubMed

    Mildner, Michael; Hacker, Stefan; Haider, Thomas; Gschwandtner, Maria; Werba, Gregor; Barresi, Caterina; Zimmermann, Matthias; Golabi, Bahar; Tschachler, Erwin; Ankersmit, Hendrik Jan

    2013-01-01

    Non-healing skin ulcers are often resistant to most common therapies. Treatment with growth factors has been demonstrated to improve closure of chronic wounds. Here we investigate whether lyophilized culture supernatant of freshly isolated peripheral blood mononuclear cells (PBMC) is able to enhance wound healing. PBMC from healthy human individuals were prepared and cultured for 24 hours. Supernatants were collected, dialyzed and lyophilized (SEC(PBMC)). Six mm punch biopsy wounds were set on the backs of C57BL/6J-mice and SEC(PBMC) containing emulsion or controls were applied daily for three days. Morphology and neo-angiogenesis were analyzed by H&E-staining and CD31 immuno-staining, respectively. In vitro effects on diverse skin cells were investigated by migration assays, cell cycle analysis, and tube formation assay. Signaling pathways were analyzed by Western blot analysis. Application of SEC(PBMC) on 6 mm punch biopsy wounds significantly enhanced wound closure. H&E staining of the wounds after 6 days revealed that wound healing was more advanced after application of SEC(PBMC) containing emulsion. Furthermore, there was a massive increase in CD31 positive cells, indicating enhanced neo-angiogenesis. In primary human fibroblasts (FB) and keratinocytes (KC) migration but not proliferation was induced. In endothelial cells (EC) SEC(PBMC) induced proliferation and tube-formation in a matrigel-assay. In addition, SEC(PBMC) treatment of skin cells led to the induction of multiple signaling pathways involved in cell migration, proliferation and survival. In summary, we could show that emulsions containing the secretome of PBMC derived from healthy individuals accelerates wound healing in a mouse model and induce wound healing associated mechanisms in human primary skin cells. The formulation and use of such emulsions might therefore represent a possible novel option for the treatment of non-healing skin ulcers.

  13. Peripheral blood neutrophil cytokine hyper-reactivity in chronic periodontitis.

    PubMed

    Ling, Martin R; Chapple, Iain L C; Matthews, John B

    2015-10-01

    Pro-inflammatory cytokine release (IL-8, IL-6, TNF-α, IL-1β) by peripheral blood neutrophils, isolated from periodontitis patients (before/after therapy) and matched controls, was determined after 18 h culture in the presence/absence of Escherichia coli LPS, opsonised Staphylococcus aureus, heat-killed Fusobacterium nucleatum and Porphyromonas gingivalis. All cultures demonstrated differences in the amounts of each cytokine detected (P < 0.0001), with a clear release pattern (IL-8 > IL-6 > TNF-α = IL-1β). Median cytokine release from unstimulated patient neutrophils was consistently, but non-significantly, higher than from control cells. Stimulated cytokine release from untreated patient neutrophils was also consistently higher than from control cells. This hyper-reactivity was significant for all tested cytokines when data for all stimuli were combined (P < 0.016). In terms of individual stimuli, significant hyper-reactivity was detected with LPS (IL-8), F. nucleatum (IL-8, TNF-α), opsonised S. aureus (IL-8, TNF-α, IL-1β) and P. gingivalis (IL-8, IL-1β). Cytokine production by patient neutrophils did not reduce following successful non-surgical periodontal therapy and, except for responses to F. nucleatum, the cytokine hyper-reactivity detected pre-therapy was retained. These data demonstrate that chronic periodontitis is characterised by neutrophils that constitutively exhibit cytokine hyper-reactivity, the effects of which could modulate local and systemic inflammatory-immune responses and influence the risk and severity of periodontitis-associated systemic inflammatory diseases.

  14. Peripheral blood cytokine and chemokine profiles in juvenile localized scleroderma

    PubMed Central

    Torok, Kathryn S.; Kurzinski, Katherine; Kelsey, Christina; Yabes, Jonathan; Magee, Kelsey; Vallejo, Abbe N.; Medsger, Thomas; Feghali-Bostwick, Carol A.

    2015-01-01

    Objective To evaluate peripheral blood T-helper (TH) cell associated cytokine and chemokine profiles in localized scleroderma (LS), and correlate them with clinical disease features, including disease activity parameters. Methods A 29-plex Luminex platform was used to analyze the humoral profile of plasma samples from 69 pediatric LS patients and 71 healthy pediatric controls. Cytokine/chemokine levels were compared between these two groups and within LS patients, focusing on validated clinical outcome measures of disease activity and damage in LS. Results Plasma levels of IP-10, MCP-1, IL-17a, IL-12p70, GM-CSF, PDGF-bb, IFN-α2, and IFN-γ were significantly higher in LS compared to healthy controls. Analysis within the LS group demonstrated IP-10, TNF-α and GM-CSF correlated with clinical measures of disease activity. Several cytokines/chemokines correlated with anti-histone antibody, while only a few correlated with positive ANA and single-stranded DNA antibody. Conclusion This is the first time that multiple cytokines and chemokines have been examined simultaneously LS. In general, a TH-1 (IFN-γ) and TH-17 (IL-17a) predominance was demonstrated in LS compared to healthy controls. There is also an IFN–γ signature with elevated IP-10, MCP-1 and IFN-γ, which has been previously demonstrated in systemic sclerosis, suggesting a shared pathophysiology. Within the LS patients, those with active disease demonstrated IP-10, TNF-α and GM-CSF, which may potentially serve as biomarkers of disease activity in the clinical setting. PMID:26254121

  15. Peripheral blood pressure by Dinamap and central blood pressure by applanation tonometry in outpatient general practice.

    PubMed

    Santiago, Luiz Miguel; Simões, Ana Rita; Ricardo Miranda, Paula; Matias, Catarina; Rosendo, Inês; Constantino, Liliana; Santos, Tiago; Neto, Maria da Glória; Francisco, Maria dos Prazeres

    2013-06-01

    Central blood pressure (CBP) is the pressure exerted by the blood column at any given moment on the aortic and carotid artery walls, which is a close proxy for the blood pressure inside the brain and the heart, and is thus a better marker of cardiovascular morbidity and mortality than peripheral blood pressure (PBP). To assess how the augmentation index (AI), peripheral pulse pressure (pPP), central pulse pressure (cPP) and subendocardial viability ratio (SEVR) vary in hypertensive patients according to level of control of CBP and PBP. We performed an observational, cross-sectional study in a convenience sample from a general practice in Central Portugal over a period of four days in May 2010. Measurements were taken after a four-minute resting period. The following values were considered to reflect controlled pressures: PBP <140/90 mmHg, CBP <130/80 mmHg, pPP <55 mmHg and cPP <45 mmHg. The sample included 92 patients, 38 male (41.3%), mean age 62.3±11.1 years, with no significant difference in gender distribution. PBP was controlled in 55 (59.8%), and CBP in 53 (57.6%). Both PBP and CBP were controlled in 50 patients (54.3%) and neither was controlled in 34 (37.9%). pPP and cPP were significantly lower in those with controlled PBP (p<0.001) and CBP (p<0.001). AI was non-significantly lower in those with controlled PBP (78±9 vs. 80.7) and those with controlled CBP (78±9 vs.81±7) (p=0.02). SEVR was within the desirable range in 92 patients (92.2%). 78.4% of individuals were taking drugs acting on the renin angiotensin aldosterone system (RAAS). In a convenience sample of 92 patients, PBP and CBP were controlled in 59.8% and 57.6%, respectively. Those with controlled PBP had significantly better peripheral systolic and diastolic blood pressure, CBP, pPP and cPP; the same was true of those with controlled CBP, who also had a significantly better AI. The percentage of the cardiac cycle in diastole had a desirable value for 92,2% of the subjects. Copyright © 2011

  16. Phagocytosis of bovine blood and milk polymorphonuclear leukocytes after ozone gas administration in vitro.

    PubMed

    Ducusin, Rio John T; Nishimura, Masakazu; Sarashina, Takao; Uzuka, Yuji; Tanabe, Shigeyuki; Otani, Masayuki

    2003-04-01

    To determine the effects of ozone on the phagocytosis of bovine polymorphonuclear leukocytes (PMNs), ozone gas was administered in vitro on the blood and milk of healthy lactating cows, cows with acute mastitis, and cows with milk fever. In the blood of healthy dairy cattle, although there was no significant effect of ozone gas on the viability of the leukocytes, phagocytosis of PMNs significantly decreased. In contrast, ozone gas administration in vitro significantly increased phagocytosis of PMNs from the blood of cows with acute mastitis and milk fever, and from mastitic milk. These findings showed that ozone administration in vitro has positive and negative effects on bovine PMN phagocytosis, depending on the health status of the animal.

  17. [Allogeneic hematopoietic transplantation with stem cells extracted from peripheral blood].

    PubMed

    Kusminsky, G; Foncuberta, M C; Aversa, L; Drelichman, G; Freigeiro, D; Burgos, R; Irrazabal, C; Gonzalez, G; Dictar, M; Niborski, R; Kohan, A; Sanchez Avalos, J C

    2000-01-01

    Fifty three patients (pts) received an allogeneic hematopoietic transplant using peripheral blood progenitor cells (PBPC). Diagnosis were acute myeloid leukemia (AML) in 16 pts, acute lymphoblastic leukemia (ALL) in 15, chronic myeloid leukemia (CML) in first chronic phase in 12, aplastic anemia in 4, myelodysplasia in 3 and Hodgkin's disease, major thalasemia and Hunter's syndrome in one each. Mean age was 20 years-old (2-55), 28 males and 25 females. Conditioning regimens were total body irradiation with 1200 cGy and cyclophosphamide 120 mg/kg in 38 pts, busulfan 16 mg/kg and cyclophosphamide 120 mg/kg in 10 pts, total lymphoid irradiation and cyclophosphamide in 3, 2 pts received other chemotherapy based conditionings. PBPC were infused unmanipulated through a central catheter. Graft versus host disease (GVHD) prophylaxis was cyclosporin and short course methotrexate. Donors were 6/6 HLA compatible siblings in 52 cases and 5/6 match in one case. PBPC mobilization was done with G-CSF at a dose of 10 micrograms/kg/day subcutaneously for four days, pheresis started on day 5. Bone marrow harvest was also done in the first thirty cases. Mean cellularities for CD34, CD3, CD4, CD8, CD56, CD19 (cel x 10(6)/kg) were 4.12; 4.59; 2.57; 1.9; 0.55 and 0.68, respectively. Mean recovery of neutrophils > 500/microL was obtained on day +11 and platelets > 20,000/microL on day +13. Patients were hospitalized for a mean period of 26 days (range 18-39) and days with parenteral antibiotics were 12.2 (5-45). Two pts had venoocclusive disease of the liver. Transplant related mortality was 15%. Acute graft versus host disease (GVHD) was observed in 43.4% of pts, only 5 pts had acute GVHD III or IV. Mean time for aGVHD diagnosis was +23 (8-76). Forty three pts were evaluable for chronic GVHD with a mean follow-up of 18 months (4-39). Chronic GVHD was observed in 26.4% by day +240, only 2 pts developed severe cGVHD. The present experience demonstrates an acceptable incidence for c

  18. Identification of interleukin-2 in human peripheral blood eosinophils.

    PubMed Central

    Levi-Schaffer, F; Barkans, J; Newman, T M; Ying, S; Wakelin, M; Hohenstein, R; Barak, V; Lacy, P; Kay, A B; Moqbel, R

    1996-01-01

    Interleukin-2 (IL-2) is an essential growth factor for T cells. Previous studies have shown that human peripheral eosinophils respond to IL-2 in chemotaxis and express the IL-2 receptor (CD25). In addition, eosinophils have been shown to transcribe messenger RNA for IL-2. The aim of the present study was to determine whether eosinophils translate mRNA for IL-2 and to determine the site of intracellular localization. By immunocytochemistry, an average of 9% of cells showed cytoplasmic staining for IL-2 in freshly isolated unstimulated blood eosinophils obtained from asthmatic subjects who were not receiving oral corticosteroid treatment (n = 5). Freshly isolated, disrupted, highly purified eosinophils (> 99%, by CD16- immunomagnetic selection) contained an average of 6 pg/10(6) cells of IL-2 measured by a specific enzyme linked immunosorbent assay (ELISA) (n = 7). Purified eosinophil incubated with serum-coated Sephadex beads showed an increase in the amount of intracellularly-retained IL-2 (26.2 +/- 7.2 pg/10(6) cells) with some evidence for release of this cytokine but only in three out of six eosinophil preparations (range 1.3-5.8 pg/10(6) cells). The intracellular localization of IL-2 was determined by fractionation of the cells on a linear (0-45%) Nycodenz gradient in sucrose buffer followed by detection of IL-2 in the fractions using an IL-2-specific ELISA and dot blotting. The majority of the IL-2 detected co-eluted with known eosinophil granule markers (i.e. major basic protein (MBP), eosinophil cationic protein (ECP), eosinophil peroxidase (EPO) and beta-hexosaminidase) but small quantities were also detected in the cytosolic (lactate dehydrogenase-(LDH) associated) and membrane (CD9+) fractions. Immunogold labelling of intact eosinophils using an anti-IL-2 monoclonal antibody confirmed IL-2 immunoreactivity in association with the eosinophil crystalline granule cores. These data are consistent with the hypothesis that eosinophils synthesize, release and

  19. Assessment of critical parameters in blood processing for the bovine interferon-gamma ELISPOT assay to detect Mycobacterium bovis infected cattle in India.

    PubMed

    Veerasami, Maroudam; Moorthy, Loganathan; Das, Dipankar; Parthasarathy, Sugumar; Chandran, Dev; Lingala, Rajendra; Srinivasan, Villuppanoor Alwar

    2011-01-01

    In vitro production of bovine interferon gamma (BoIFN-γ) cytokine from bovine peripheral blood mononuclear cells (PBMCs) can be detected using the most sensitive enzyme-linked immunosorbent spot (ELISPOT) assay. ELISPOT assays are dependent on the quantity and quality of PBMC preparations and hence contribute significantly to the performance of this assay. In order to standardise the methods for isolation of PBMCs, we compared two methods for the processing of bovine blood which included aliquots of blood that were stored in a horizontal position without dilution or agitation and aliquots of blood that were immediately diluted 1:1 with complete Rosewell Park Memorial Institute (RPMI) 1640 medium and stored in a horizontal position with gentle agitation. PBMCs were isolated at 2, 4, 6, 8 and 24 h and at 4°C and at 22°C ± 2°C. They were stimulated using tuberculosis-specific antigens, after which the ELISPOT assay was performed. Quantities of spot-forming cells (SFC) created by the release of BoIFN-γ in ELISPOT assays were significantly greater in the samples stored at 22°C ± 2°C than those held +4°C and the intensity of the signals dropped following processing after 6 h. A further drop in SFC was observed in those samples that had been stored undiluted and without agitation. These findings demonstrated that optimisation of PBMC isolation procedures can lead to increased sensitivity in the detection of BoIFN-γ using the ELISPOT assay, thus contributing to an enhanced diagnosis of bovine tuberculosis.

  20. Activity of histidine in peripheral blood erythrocytes of pregnant women during exacerbation of cytomegalovirus infection.

    PubMed

    Lutsenko, M T; Andrievskaya, I A

    2014-10-01

    We studied the effect of active cytomegalovirus infection on histidine content in peripheral blood erythrocytes of pregnant women at gestation weeks 20-22 and its involvement into hemoglobin oxygenation. Using the histochemical technique developed by us, we studied the distribution of products of specific reaction for histidine in peripheral blood erythrocytes of pregnant women. The percentage of histidine-positive erythrocytes and their area were evaluated. The relationship between the distribution of the products of the reaction for histidine in peripheral blood erythrocytes of pregnant women and the titer of anti-cytomegalovirus IgG was revealed. The histidine content in peripheral blood erythrocytes of pregnant women with active cytomegalovirus infection was reduced, which impaired heme binding to globin and decreased the formation of oxyhemoglobin.

  1. Growth of erythroid burst-forming units (BFU-E) in cultures of canine bone marrow and peripheral blood cells: effect of serum from irradiated dogs

    SciTech Connect

    Kreja, L.; Baltschukat, K.; Nothdurft, W.

    1988-08-01

    Erythroid burst-forming units (BFU-E) from canine bone marrow and peripheral blood could be grown in methylcellulose in the presence of an appropriate batch of fetal calf serum (FCS), transferrin, and erythropoietin (Epo). However, improved colony formation (size and number of bursts) was obtained when serum from total body irradiated dogs was present in the culture. This serum, obtained from dogs at day 9 after total body irradiation with a dose of 3.9 Gy, reduced markedly the Epo requirement of BFU-E. Furthermore, it allowed the omission of FCS from the culture medium if cholesterol and bovine serum albumin (BSA) were used as FCS substitutes. BFU-E concentrations were found to be rather different in the peripheral blood and in bone marrow samples from different sites (i.e., iliac crest, sternum, and humerus) of normal beagles. The studies further show that canine bone marrow BFU-E can be cryopreserved in liquid nitrogen.

  2. Simple Radiowave-Based Method For Measuring Peripheral Blood Flow Project

    NASA Technical Reports Server (NTRS)

    Oliva-Buisson, Yvette J.

    2014-01-01

    Project objective is to design small radio frequency based flow probes for the measurement of blood flow velocity in peripheral arteries such as the femoral artery and middle cerebral artery. The result will be the technological capability to measure peripheral blood flow rates and flow changes during various environmental stressors such as microgravity without contact to the individual being monitored. This technology may also lead to an easier method of detecting venous gas emboli during extravehicular activities.

  3. [Effect of stevia on the picture of peripheral blood under exposure to vibration].

    PubMed

    Adamyan, Ts I; Gevorkyan, E S

    2014-01-01

    There were investigated changes in the peripheral blood of rabbits under prolonged exposure to vibration (5, 10, 20, 30 days). In a separate series of experiments, the nature of changes in the peripheral blood was investigated under the combined action of vibration and stevia leaves. Contained in stevia biologically active substances were found to accelerate metabolism in bone marrow stem cells, promote the compensatory ability of the organism, thereby providing the resistance of the body to the vibration factor.

  4. Plasma histamine concentration and histamine detection in peripheral blood eosinophils in cats.

    PubMed

    Kadoya, Michiyo; Momoi, Yasuyuki; Iwasaki, Toshiroh

    2006-10-01

    Plasma histamine levels were measured in 11 clinically healthy cats and 15 cats with allergic dermatitis. Histamine levels were markedly elevated in 5/15 allergic cats. A calcium ionophore, A23187, stimulates histamine release from feline peripheral blood cells. Immunostaining of blood smears from clinically healthy cats revealed that approximately 10% of eosinophils possessed histamine-containing granules. These results indicate that some peripheral eosinophils in cats contain histamine and can release histamine by appropriate stimulation.

  5. Determination of testosterone esters and estradiol esters in bovine and porcine blood serum.

    PubMed

    Rejtharová, Martina; Rejthar, Libor; Čačková, Katarína

    2017-04-01

    Monitoring of steroid esters in blood serum is desirable in order to detect the possible illegal use of natural hormones as growth promoters. A method for the determination of testosterone propionate, testosterone benzoate, testosterone isocaproate, testosterone decanoate and estradiol benzoate in bovine and porcine blood serum was developed. The procedure consists of protein precipitation and removal of phospholipids using a HybridSPE®-Phospholipid column followed by clean-up on a hydrophilic modified styrene polymer Supel(TM)-Select HLB column and LC-MS/MS measurement. The method was validated according to Commission Decision 2002/657/EC. Decision limits for all analytes were observed in the range 5-30 pg ml(-)(1). The method was shown to be robust for bovine and porcine serum analyses and can be applied for both screening and confirmatory determination in routine residue monitoring.

  6. Cytomegalovirus infection after allogeneic transplantation: comparison of cord blood with peripheral blood and marrow graft sources.

    PubMed

    Walker, Christopher M; van Burik, Jo-Anne H; De For, Todd E; Weisdorf, Daniel J

    2007-09-01

    Cytomegalovirus (CMV) infection is an important complication following allogeneic hematopoietic stem cell transplant (HSCT), but the natural history in the cord blood setting has not been well studied. We assessed CMV infection episodes in 753 consecutive allogeneic HSCT recipients at the University of Minnesota between January 1, 1998 and December 31, 2003. The 6-month cumulative incidence of viremia/antigenemia was 22% by day +182: 21% (95% confidence interval 16%-26%) in cord blood recipients (UCB), 24% (20%-28%) in marrow (BM), and 22% (16%-28%) using peripheral blood grafts (PBSC). CMV disease incidence was 6% (2%-10%) in UCB, 8% (5%-11%) in BM, and 9% (6%-12%) in PBSC. In multivariate analysis, CMV infection (viremia/antigenemia and disease) was significantly more likely in patients who were seropositive to CMV, in those with acute graft versus host disease, and in those receiving T cell-depleted grafts. Graft source did not independently contribute to the risk of CMV infection and did not impact survival after CMV infection. These data confirm that recipient CMV serostatus remains the dominant risk factor for CMV infection. Recipients of UCB have similar risks of CMV infection, responses to antiviral therapy, and survival following CMV infection as recipients of either marrow or PBSC.

  7. Blood Cultures at Central Line Insertion in the Intensive Care Unit: Comparison with Peripheral Venipuncture▿

    PubMed Central

    Stohl, Sheldon; Benenson, Shmuel; Sviri, Sigal; Avidan, Alexander; Block, Colin; Sprung, Charles L.; Levin, Phillip D.

    2011-01-01

    Blood cultures are a key diagnostic test for intensive care unit (ICU) patients; however, contaminants complicate interpretations and lead to unnecessary antibiotic administration and costs. Indications for blood cultures and central venous catheter (CVC) insertions often overlap for ICU patients. Obtaining blood cultures under the strict sterile precautions utilized for CVC insertion might be expected to decrease culture contamination. This retrospective study compared the results of blood cultures taken at CVC insertion, at arterial line insertion, and from peripheral venipuncture in order to validate the advantage of CVC insertion cultures. Cultures from indwelling lines were excluded. Results of 14,589 blood cultures, including 2,736 (19%) CVC, 1,513 (10%) arterial line, and 10,340 (71%) peripheral cultures taken over 5.5 years in two ICUs (general and medical) were analyzed. CVC cultures were contaminated more frequently than arterial line or peripheral cultures (225/2,736 [8%] CVC, 48/1,513 [3%] arterial line, and 378/10,340 (4%) peripheral cultures [P < 0.001 for CVC versus peripheral and CVC versus arterial line cultures]). True pathogens were found more frequently in CVC insertion cultures (334/2,736 [12%] CVC, 155/1,513 [10%] arterial line, and 795/10,340 [8%] peripheral cultures [P < 0.001 for CVC versus peripheral cultures; P = 0.055 for CVC versus arterial line cultures; P < 0.001 for peripheral versus arterial line cultures]). Contamination and true-positive rates were similar for culture sets from the two ICUs for each given culture source. Despite superior sterile precautions, cultures taken at the time of central line insertion had a higher contamination rate than did either peripheral or arterial line blood cultures. This may be related to the increased manipulations required for CVC insertion. PMID:21525219

  8. Gastrin in portal and peripheral venous blood after feeding in man

    PubMed Central

    Dencker, H.; Håkanson, R.; Liedberg, G.; Norryd, C.; Oscarson, J.; Rehfeld, J. F.; Stadil, F.

    1973-01-01

    The concentrations of immunoreactive gastrin in serum from portal and peripheral venous blood were determined in 10 patients with indwelling portal catheters before and after feeding. No significant differences were found between the gastrin concentrations in portal and peripheral serum. Gel filtration studies of serum did not reveal any differences between the gastrin components of portal and peripheral venous serum. Since neither the concentrations of immunoreactive gastrin nor the four gastrin components differed between portal and peripheral serum it is suggested that the liver is without effect on gastrin metabolism. PMID:4761604

  9. A color and shape based algorithm for segmentation of white blood cells in peripheral blood and bone marrow images.

    PubMed

    Arslan, Salim; Ozyurek, Emel; Gunduz-Demir, Cigdem

    2014-06-01

    Computer-based imaging systems are becoming important tools for quantitative assessment of peripheral blood and bone marrow samples to help experts diagnose blood disorders such as acute leukemia. These systems generally initiate a segmentation stage where white blood cells are separated from the background and other nonsalient objects. As the success of such imaging systems mainly depends on the accuracy of this stage, studies attach great importance for developing accurate segmentation algorithms. Although previous studies give promising results for segmentation of sparsely distributed normal white blood cells, only a few of them focus on segmenting touching and overlapping cell clusters, which is usually the case when leukemic cells are present. In this article, we present a new algorithm for segmentation of both normal and leukemic cells in peripheral blood and bone marrow images. In this algorithm, we propose to model color and shape characteristics of white blood cells by defining two transformations and introduce an efficient use of these transformations in a marker-controlled watershed algorithm. Particularly, these domain specific characteristics are used to identify markers and define the marking function of the watershed algorithm as well as to eliminate false white blood cells in a postprocessing step. Working on 650 white blood cells in peripheral blood and bone marrow images, our experiments reveal that the proposed algorithm improves the segmentation performance compared with its counterparts, leading to high accuracies for both sparsely distributed normal white blood cells and dense leukemic cell clusters. © 2014 International Society for Advancement of Cytometry.

  10. Cord blood T cells mediate enhanced antitumor effects compared with adult peripheral blood T cells.

    PubMed

    Hiwarkar, Prashant; Qasim, Waseem; Ricciardelli, Ida; Gilmour, Kimberly; Quezada, Sergio; Saudemont, Aurore; Amrolia, Persis; Veys, Paul

    2015-12-24

    Unrelated cord blood transplantation (CBT) without in vivo T-cell depletion is increasingly used to treat high-risk hematologic malignancies. Following T-replete CBT, naïve CB T cells undergo rapid peripheral expansion with memory-effector differentiation. Emerging data suggest that unrelated CBT, particularly in the context of HLA mismatch and a T-replete graft, may reduce leukemic relapse. To study the role of CB T cells in mediating graft-versus-tumor responses and dissect the underlying immune mechanisms for this, we compared the ability of HLA-mismatched CB and adult peripheral blood (PB) T cells to eliminate Epstein-Barr virus (EBV)-driven human B-cell lymphoma in a xenogeneic NOD/SCID/IL2rg(null) mouse model. CB T cells mediated enhanced tumor rejection compared with equal numbers of PB T cells, leading to improved survival in the CB group (P < .0003). Comparison of CB T cells that were autologous vs allogeneic to the lymphoma demonstrated that this antitumor effect was mediated by alloreactive rather than EBV-specific T cells. Analysis of tumor-infiltrating lymphocytes demonstrated that CB T cells mediated this enhanced antitumor effect by rapid infiltration of the tumor with CCR7(+)CD8(+) T cells and prompt induction of cytotoxic CD8(+) and CD4(+) T-helper (Th1) T cells in the tumor microenvironment. In contrast, in the PB group, this antilymphoma effect is impaired because of delayed tumoral infiltration of PB T cells and a relative bias toward suppressive Th2 and T-regulatory cells. Our data suggest that, despite being naturally programmed toward tolerance, reconstituting T cells after unrelated T-replete CBT may provide superior Tc1-Th1 antitumor effects against high-risk hematologic malignancies.

  11. Morphological changes of red blood cells in peripheral blood smear of patients with pregnancy-related hypertensive disorders.

    PubMed

    Hernández Hernández, Jorge Daniel; Villaseñor, Onésimo Rangel; Del Rio Alvarado, Javier; Lucach, Roberto Ortega; Zárate, Arturo; Saucedo, Renata; Hernández-Valencia, Marcelino

    2015-08-01

    Pregnancy-related hypertensive disorders are complications in which risk factors are identified such as nulliparity, age, malnutrition, obesity and social issues. Those statements are explained by theories of abnormal placentation, immunological inadequacy, genetics and oxidative stress, but all theories converge in endothelial damage, which is able to mechanically deform and hemolyze erythrocytes as they pass through the capillaries. Given the effects of endothelial damage, the aim of the study was to determine erythrocyte alterations in peripheral blood smear of patients with hypertensive disorders of pregnancy that could be used as prognostic condition. We performed a prospective, descriptive and observational study where all patients with hypertensive disorders admitted to the obstetrics and gynecology service of a specialty hospital were recruited. Patients who provided signed informed consent underwent peripheral blood smear. Results were tabulated in percentage graphics and analyzed with Cramer's V based on χ(2). The peripheral blood smear consisted of an extended drop of peripheral blood from the patient with subsequent hematological staining done with Romanowsky stain. A total of 119 samples were analyzed; 74% showed abnormal morphology of erythrocytes and the most frequent abnormality was the presence of schistocytes in up to 39% of samples. Descriptive analysis showed a degree of association to independent variables with Cramer's V = 0.41 value (p <0.05). A high percentage of patients with hypertensive disorders of pregnancy show some morphologic alterations of erythrocytes in peripheral blood smear. Copyright © 2015 IMSS. Published by Elsevier Inc. All rights reserved.

  12. Bovine whole-blood culture as a tool for the measurement of endotoxin activities in Gram-negative bacterial vaccines.

    PubMed

    Imamura, Saiki; Nakamizo, Mari; Kawanishi, Michiko; Nakajima, Nao; Yamamoto, Kinya; Uchiyama, Mariko; Hirano, Fumiya; Nagai, Hidetaka; Kijima, Mayumi; Ikebuchi, Ryoyo; Mekata, Hirohisa; Murata, Shiro; Konnai, Satoru; Ohashi, Kazuhiko

    2013-05-15

    In order to analyze bovine immune reactions against the Gram-negative bacterial vaccine, bovine whole-blood culture was used to investigate the pro-inflammatory cytokine responses stimulated with lipopolysaccharides (LPS) extracted from Escherichia coli, Salmonella enterica, Pseudomonas aeruginosa, and Klebsiella pneumoniae. We also examined the interaction between LPS and aluminum hydroxide gel for endotoxin activity and pro-inflammatory cytokine responses of whole bovine blood. Alteration in the mRNA concentrations of tumor necrosis factor (TNF)-α, interleukin (IL)-1β, and IL-10 in whole-blood culture at 4h after stimulation with different doses of LPS was observed and determined by quantitative reverse-transcription polymerase chain reaction (qRT-PCR). The mRNA concentrations of TNF-α and IL-1β changed in a dose-dependent manner and differed depending on the type of LPS. Limulus test revealed that endotoxin activity was remarkably reduced when aluminum hydroxide gel was added to LPS. In contrast, the mRNA concentration of TNF-α in whole bovine blood was enhanced by LPS mixed with aluminum hydroxide gel. These results suggest that bovine whole-blood culture can be utilized to detect endotoxin activity of Gram-negative bacterial vaccines. In addition, whole-blood culture offers several advantages, such as ease of performance, few preparation artifacts, and a physiological cell environment, for investigating bovine immune response compared with the Limulus test.

  13. Blood-informative transcripts define nine common axes of peripheral blood gene expression.

    PubMed

    Preininger, Marcela; Arafat, Dalia; Kim, Jinhee; Nath, Artika P; Idaghdour, Youssef; Brigham, Kenneth L; Gibson, Greg

    2013-01-01

    We describe a novel approach to capturing the covariance structure of peripheral blood gene expression that relies on the identification of highly conserved Axes of variation. Starting with a comparison of microarray transcriptome profiles for a new dataset of 189 healthy adult participants in the Emory-Georgia Tech Center for Health Discovery and Well-Being (CHDWB) cohort, with a previously published study of 208 adult Moroccans, we identify nine Axes each with between 99 and 1,028 strongly co-regulated transcripts in common. Each axis is enriched for gene ontology categories related to sub-classes of blood and immune function, including T-cell and B-cell physiology and innate, adaptive, and anti-viral responses. Conservation of the Axes is demonstrated in each of five additional population-based gene expression profiling studies, one of which is robustly associated with Body Mass Index in the CHDWB as well as Finnish and Australian cohorts. Furthermore, ten tightly co-regulated genes can be used to define each Axis as "Blood Informative Transcripts" (BITs), generating scores that define an individual with respect to the represented immune activity and blood physiology. We show that environmental factors, including lifestyle differences in Morocco and infection leading to active or latent tuberculosis, significantly impact specific axes, but that there is also significant heritability for the Axis scores. In the context of personalized medicine, reanalysis of the longitudinal profile of one individual during and after infection with two respiratory viruses demonstrates that specific axes also characterize clinical incidents. This mode of analysis suggests the view that, rather than unique subsets of genes marking each class of disease, differential expression reflects movement along the major normal Axes in response to environmental and genetic stimuli.

  14. Blood-Informative Transcripts Define Nine Common Axes of Peripheral Blood Gene Expression

    PubMed Central

    Preininger, Marcela; Arafat, Dalia; Kim, Jinhee; Nath, Artika P.; Idaghdour, Youssef; Brigham, Kenneth L.; Gibson, Greg

    2013-01-01

    We describe a novel approach to capturing the covariance structure of peripheral blood gene expression that relies on the identification of highly conserved Axes of variation. Starting with a comparison of microarray transcriptome profiles for a new dataset of 189 healthy adult participants in the Emory-Georgia Tech Center for Health Discovery and Well-Being (CHDWB) cohort, with a previously published study of 208 adult Moroccans, we identify nine Axes each with between 99 and 1,028 strongly co-regulated transcripts in common. Each axis is enriched for gene ontology categories related to sub-classes of blood and immune function, including T-cell and B-cell physiology and innate, adaptive, and anti-viral responses. Conservation of the Axes is demonstrated in each of five additional population-based gene expression profiling studies, one of which is robustly associated with Body Mass Index in the CHDWB as well as Finnish and Australian cohorts. Furthermore, ten tightly co-regulated genes can be used to define each Axis as “Blood Informative Transcripts” (BITs), generating scores that define an individual with respect to the represented immune activity and blood physiology. We show that environmental factors, including lifestyle differences in Morocco and infection leading to active or latent tuberculosis, significantly impact specific axes, but that there is also significant heritability for the Axis scores. In the context of personalized medicine, reanalysis of the longitudinal profile of one individual during and after infection with two respiratory viruses demonstrates that specific axes also characterize clinical incidents. This mode of analysis suggests the view that, rather than unique subsets of genes marking each class of disease, differential expression reflects movement along the major normal Axes in response to environmental and genetic stimuli. PMID:23516379

  15. Colloidal Properties of Nanoerythrosomes Derived from Bovine Red Blood Cells.

    PubMed

    Kuo, Yuan-Chia; Wu, Hsuan-Chen; Hoang, Dao; Bentley, William E; D'Souza, Warren D; Raghavan, Srinivasa R

    2016-01-12

    Liposomes are nanoscale containers that are typically synthesized from lipids using a high-shear process such as extrusion or sonication. While liposomes are extensively used in drug delivery, they do suffer from certain problems including limited colloidal stability and short circulation times in the body. As an alternative to liposomes, we explore a class of container structures derived from erythrocytes (red blood cells). The procedure involves emptying the inner contents of these cells (specifically hemoglobin) and resuspending the empty structures in buffer, followed by sonication. The resulting structures are termed nanoerythrosomes (NERs), i.e., they are membrane-covered nanoscale containers, much like liposomes. Cryo-transmission electron microscopy (cryo-TEM) and small-angle neutron scattering (SANS) are employed for the first time to study these NERs. The results reveal that the NERs are discrete spheres (∼110 nm diameter) with a unilamellar membrane of thickness ∼4.5 nm. Remarkably, the biconcave disc-like shape of erythrocytes is also exhibited by the NERs under hypertonic conditions. Moreover, unlike typical liposomes, NERs show excellent colloidal stability in both buffer as well as in serum at room temperature, and are also able to withstand freeze-thaw cycling. We have explored the potential for using NERs as colloidal vehicles for targeted delivery. Much like conventional liposomes, NER membranes can be decorated with fluorescent or other markers, solutes can be encapsulated in the cores of the NERs, and NERs can be targeted to specifically bind to mammalian cells. Our study shows that NERs are a promising and versatile class of nanostructures. NERs that are harvested from a patient's own blood and reconfigured for nanomedicine can potentially offer several benefits including biocompatibility, minimization of immune response, and extended circulation time in the body.

  16. Expression of STK39 in peripheral blood of hypertension patients and the relationship between its genetic polymorphism and blood pressure.

    PubMed

    Li, B; Yang, M; Liu, J W

    2015-12-09

    This study investigated the STK39 expression in peripheral blood of hypertension patients and the relation between its genetic polymorphism and blood pressure. The observation group comprised of 42 primary hypertension patients admitted to our hospital, and the control group comprised of 30 healthy individuals who underwent physical examination in our hospital during the same period. Fasting venous blood was collected from both groups in the morning to determine the STK39 mRNA and protein levels in peripheral blood using quantitative real-time PCR and western blot. STK39 gene SNP (rs6433027) was sequenced using PCR and its genetic variation was analyzed. The relationship between STK39 protein level, genetic variation, and diastolic and systolic blood pressure was also analyzed. The observation group showed increased STK39 mRNA and protein levels in peripheral blood compared to the control group, and the difference was statistically significant (P < 0.05), suggesting C/T mutation in STK39 gene SNP (rs6433027). Correlation analysis showed positive association between STK39 protein level and diastolic and systolic blood pressure (P < 0.05), indicating a positive association between C/T genetic mutation and diastolic and systolic blood pressure (P < 0.05). In conclusion, STK39 mRNA and protein express abnormally in primary hypertension patients with genetic variation, which is related to the blood pressure.

  17. Quantification of PrPC in bovine peripheral tissues: Analysis in wild-type and PrPC-deficient cattle.

    PubMed

    Kobayashi, Shin-Ichi; Ano, Yasuhisa; Sakudo, Akikazu; Yukawa, Masayoshi; Sigiura, Katsuaki; Manabe, Noboru; Nakayama, Hiroyuki; Onodera, Takashi

    2009-01-01

    Cellular PrP (PrPC) is necessary for bovine spongiform encephalopathy (BSE) infection. The purpose of the present experiment was the quantification of PrPC in peripheral tissues to assess the risk of BSE infection from these tissues. The tissue distribution of PrPC was examined by a sandwich enzyme-linked immunosorbent assay (sELISA) and histochemical analysis. PrPC-deficient cows were used as a negative control. The sELISA revealed that the brain contained the highest PrPC content (10.7 µg/g tissue), while other organs/tissues harbored lower amounts, in decreasing order as follows: longissimus capitis muscle, iliocostalis thoracis muscle, splenius muscle, biceps femoris muscle, triceps brachii muscle, longissimus thoracis muscle, ileum, jejunum, duodenum, colon, cecum, apex linguae, omotransversarius muscle, posterior part of the corpus linguae, anterior part of the corpus linguae and radix linguae (5.2- to 31-fold less PrPC than the brain). In the tissue/organs of PrP-deficient cows, PrPC levels were under the limit of detection. Histochemical analysis showed that PrPC was expressed in nerve cells in intestinal tissues. The presence of PrPC in the bovine tongue, skeletal muscles and intestines raises the possibility of PrPSc accumulation in these tissues, indicating that these organs/tissues may serve as potential sources of BSE infection.

  18. Evaluation of bovine luteal blood flow by using color Doppler ultrasonography.

    PubMed

    Lüttgenau, J; Bollwein, H

    2014-04-01

    Since luteal vascularization plays a decisive role for the function of the corpus luteum (CL), the investigation of luteal blood flow (LBF) might give valuable information about the physiology and patho-physiology of the CL. To quantify LBF, usually Power mode color Doppler ultrasonography is used. This method detects the number of red blood cells moving through the vessels and shows them as color pixels on the B-mode image of the CL. The area of color pixels is measured with computer-assisted image analysis software and is used as a semiquantitative parameter for the assessment of LBF. Although Power mode is superior for the evaluation of LBF compared to conventional color Doppler ultrasonography, which detects the velocity of blood cells, it is still not sufficiently sensitive to detect the blood flow in the small vessels in the center of the bovine CL. Therefore, blood flow can only be measured in the bigger luteal vessels in the outer edge of the CL. Color Doppler ultrasonographic studies of the bovine estrous cycle have shown that plasma progesterone (P4) concentration can be more reliably predicted by LBF than by luteal size (LS), especially during the CL regression. During the midluteal phase, cows with low P4 level showed smaller CL, but LBF, related to LS, did not differ between cows with low and high P4 levels. In contrast to non-pregnant cows, a significant rise in LBF was observed three weeks after insemination in pregnant cows. However, LBF was not useful for an early pregnancy diagnosis due to high LBF variation among cows. When the effects of an acute systemic inflammation and exogenous hormones on the CL are examined, the LBF determination is more sensitive than LS assessment. In conclusion, color Doppler ultrasonography of the bovine CL provides additional information on luteal function compared to measurements of LS and plasma P4, but its value as a parameter concerning assessment of fertility in cows has to be clarified.

  19. Detection and enrichment of disseminated renal carcinoma cells from peripheral blood by immunomagnetic cell separation.

    PubMed

    Bilkenroth, U; Taubert, H; Riemann, D; Rebmann, U; Heynemann, H; Meye, A

    2001-05-15

    We have established an immunomagnetic separation procedure for the detection of circulating tumor cells in the peripheral blood based on the magnetic cell sorting (MACS) technique. In previous in vitro experiments, renal-cell carcinoma (RCC) cells were mixed with peripheral blood. In dilutions of 1:200 to 1:107 tumor cells per mononuclear blood cells, an average recovery rate of 84% of tumor cells was determined. In our study, 104 peripheral blood samples from 59 renal carcinoma patients were analyzed. MACS resulted in significant depletion of leukocytes, permitting a search for tumor cells on just 1 slide. Analyzing 8 ml of peripheral blood per patient, 19/59 RCC patients carried disseminated tumor cells (32%) in the range of 1 to 38 cells (median 8). Interestingly, for the cytokeratin-positive (CK+) patient group, we found a correlation between tumor cell number and grading (G2 vs. G3) and an increased number of CK+ patients with advanced tumor stage. MACS appears to be an efficient technique to detect disseminated tumor cells in peripheral blood.

  20. Platelet and red blood cell utilization and transfusion independence in umbilical cord blood and allogeneic peripheral blood hematopoietic cell transplants.

    PubMed

    Solh, Melhem; Brunstein, Claudio; Morgan, Shanna; Weisdorf, Daniel

    2011-05-01

    Allogeneic hematopoietic cell transplantation (HCT) recipients have substantial transfusion requirements. Factors associated with increased transfusions and the extent of blood product use in umbilical cord blood (UCB) recipients are uncertain. We reviewed blood product use in 229 consecutive adult recipients of allogeneic HCT at the University of Minnesota: 147 with leukemia, 82 lymphoma or myeloma; 58% received unrelated UCB and 43% sibling donor peripheral blood stem cell (PBSC) grafts. Although neutrophil recovery was prompt (UCB median 17, range: 2-45 days, and PBSC 14, range: 3-34 days), only 135 of 229 (59% cumulative incidence) achieved red blood cell (RBC) independence and 157 (69%) achieved platelet independence by 6 months. Time to platelet independence was prolonged in UCB recipients (median UCB 41 versus PBSC 14 days) and in patients who had received a prior transplant (median 48 versus 32 days). Patients who received UCB grafts required more RBC through day 60 post-HCT (mean UCB 7.8 (95% confidence interval [CI] 6.7-8.9) versus PBSC 5.2 (3.7-6.7) transfusions, P = .04), and more platelet transfusions (mean 25.2 (95% CI 22.1-28.2) versus 12.9 (9.4-16.4), P < .01) compared to PBSC recipients. Patients receiving myeloablative (MA) conditioning required more RBC and platelet transfusions during the first 2 months post-HCT compared to reduced-intensity conditioning (RIC) (7.4 versus 6.2, P = .30 for RBC; 23.2 versus 17.5, P = .07 for platelets). Despite prompt neutrophil engraftment, UCB recipients had delayed platelet recovery as well as more prolonged and costly blood product requirements. Enhanced approaches to accelerate multilineage engraftment could limit the transfusion-associated morbidity and costs accompanying UCB allotransplantation.

  1. Peripheral-blood stem cells versus bone marrow from unrelated donors.

    PubMed

    Anasetti, Claudio; Logan, Brent R; Lee, Stephanie J; Waller, Edmund K; Weisdorf, Daniel J; Wingard, John R; Cutler, Corey S; Westervelt, Peter; Woolfrey, Ann; Couban, Stephen; Ehninger, Gerhard; Johnston, Laura; Maziarz, Richard T; Pulsipher, Michael A; Porter, David L; Mineishi, Shin; McCarty, John M; Khan, Shakila P; Anderlini, Paolo; Bensinger, William I; Leitman, Susan F; Rowley, Scott D; Bredeson, Christopher; Carter, Shelly L; Horowitz, Mary M; Confer, Dennis L

    2012-10-18

    Randomized trials have shown that the transplantation of filgrastim-mobilized peripheral-blood stem cells from HLA-identical siblings accelerates engraftment but increases the risks of acute and chronic graft-versus-host disease (GVHD), as compared with the transplantation of bone marrow. Some studies have also shown that peripheral-blood stem cells are associated with a decreased rate of relapse and improved survival among recipients with high-risk leukemia. We conducted a phase 3, multicenter, randomized trial of transplantation of peripheral-blood stem cells versus bone marrow from unrelated donors to compare 2-year survival probabilities with the use of an intention-to-treat analysis. Between March 2004 and September 2009, we enrolled 551 patients at 48 centers. Patients were randomly assigned in a 1:1 ratio to peripheral-blood stem-cell or bone marrow transplantation, stratified according to transplantation center and disease risk. The median follow-up of surviving patients was 36 months (interquartile range, 30 to 37). The overall survival rate at 2 years in the peripheral-blood group was 51% (95% confidence interval [CI], 45 to 57), as compared with 46% (95% CI, 40 to 52) in the bone marrow group (P=0.29), with an absolute difference of 5 percentage points (95% CI, -3 to 14). The overall incidence of graft failure in the peripheral-blood group was 3% (95% CI, 1 to 5), versus 9% (95% CI, 6 to 13) in the bone marrow group (P=0.002). The incidence of chronic GVHD at 2 years in the peripheral-blood group was 53% (95% CI, 45 to 61), as compared with 41% (95% CI, 34 to 48) in the bone marrow group (P=0.01). There were no significant between-group differences in the incidence of acute GVHD or relapse. We did not detect significant survival differences between peripheral-blood stem-cell and bone marrow transplantation from unrelated donors. Exploratory analyses of secondary end points indicated that peripheral-blood stem cells may reduce the risk of graft failure

  2. Discovery and implementation of transcriptional biomarkers of synthetic LXR agonists in peripheral blood cells.

    PubMed

    DiBlasio-Smith, Elizabeth A; Arai, Maya; Quinet, Elaine M; Evans, Mark J; Kornaga, Tad; Basso, Michael D; Chen, Liang; Feingold, Irene; Halpern, Anita R; Liu, Qiang-Yuan; Nambi, Ponnal; Savio, Dawn; Wang, Shuguang; Mounts, William M; Isler, Jennifer A; Slager, Anna M; Burczynski, Michael E; Dorner, Andrew J; LaVallie, Edward R

    2008-10-16

    LXRs (Liver X Receptor alpha and beta) are nuclear receptors that act as ligand-activated transcription factors. LXR activation causes upregulation of genes involved in reverse cholesterol transport (RCT), including ABCA1 and ABCG1 transporters, in macrophage and intestine. Anti-atherosclerotic effects of synthetic LXR agonists in murine models suggest clinical utility for such compounds. Blood markers of LXR agonist exposure/activity were sought to support clinical development of novel synthetic LXR modulators. Transcript levels of LXR target genes ABCA1 and ABCG1 were measured using quantitative reverse transcriptase/polymerase chain reaction assays (qRT-PCR) in peripheral blood from mice and rats (following a single oral dose) and monkeys (following 7 daily oral doses) of synthetic LXR agonists. LXRalpha, LXRbeta, ABCA1, and ABCG1 mRNA were measured by qRT-PCR in human peripheral blood mononuclear cells (PBMC), monocytes, T- and B-cells treated ex vivo with WAY-252623 (LXR-623), and protein levels in human PBMC were measured by Western blotting. ABCA1/G1 transcript levels in whole-blood RNA were measured using analytically validated assays in human subjects participating in a Phase 1 SAD (Single Ascending Dose) clinical study of LXR-623. A single oral dose of LXR agonists induced ABCA1 and ABCG1 transcription in rodent peripheral blood in a dose- and time-dependent manner. Induction of gene expression in rat peripheral blood correlated with spleen expression, suggesting LXR gene regulation in blood has the potential to function as a marker of tissue gene regulation. Transcriptional response to LXR agonist was confirmed in primates, where peripheral blood ABCA1 and ABCG1 levels increased in a dose-dependent manner following oral treatment with LXR-623. Human PBMC, monocytes, T- and B cells all expressed both LXRalpha and LXRbeta, and all cell types significantly increased ABCA1 and ABCG1 expression upon ex vivo LXR-623 treatment. Peripheral blood from a

  3. Cationic pyridinium porphyrins appending different peripheral substituents: Spectroscopic studies on their interactions with bovine serum albumin

    NASA Astrophysics Data System (ADS)

    Zhao, Ping; Huang, Jin-Wang; Ji, Liang-Nian

    2012-03-01

    The interaction of cationic pyridinium porphyrins appending methylpyridyl, hydroxyphenyl, propionoxyphenyl or carboxyphenyl group at meso-20-position of porphyrin core with bovine serum albumin (BSA), was studied by the combination of absorption spectroscopy, surface-enhanced Raman spectroscopy (SERS), circular dichroism (CD) spectroscopy, fluorescence spectroscopy and synchronous spectroscopy. The spectral monitoring results indicate that the studied compounds could bind with the BSA molecule and the calculated binding constants show that the tetracationic porphyrin has higher binding affinity than those tricationic ones. The interactions between porphyrins and BSA employ an electrostatic binding mechanism and there was only one binding site which located on the surface of the protein molecule.

  4. Raman spectroscopy coupled with advanced statistics for differentiating menstrual and peripheral blood.

    PubMed

    Sikirzhytskaya, Aliaksandra; Sikirzhytski, Vitali; Lednev, Igor K

    2014-01-01

    Body fluids are a common and important type of forensic evidence. In particular, the identification of menstrual blood stains is often a key step during the investigation of rape cases. Here, we report on the application of near-infrared Raman microspectroscopy for differentiating menstrual blood from peripheral blood. We observed that the menstrual and peripheral blood samples have similar but distinct Raman spectra. Advanced statistical analysis of the multiple Raman spectra that were automatically (Raman mapping) acquired from the 40 dried blood stains (20 donors for each group) allowed us to build classification model with maximum (100%) sensitivity and specificity. We also demonstrated that despite certain common constituents, menstrual blood can be readily distinguished from vaginal fluid. All of the classification models were verified using cross-validation methods. The proposed method overcomes the problems associated with currently used biochemical methods, which are destructive, time consuming and expensive.

  5. The utility of peripheral blood smear review for identifying specimens for flow cytometric immunophenotyping.

    PubMed

    Craig, F E

    2017-05-01

    Laboratory professionals are in an ideal situation to identify CBC and peripheral blood smear findings that raise the possibility of a hematolymphoid neoplasm, and based on this information make recommendations for additional studies, such as flow cytometric immunophenotyping. In some circumstances a definitive diagnosis can be established from the combined peripheral blood morphologic and immunophenotypic findings obviating the need for bone marrow evaluation, such as for chronic lymphocytic leukemia. Occasionally flow cytometric studies are superior to morphologic assessment, such as in screening for hairy cell leukemia or identifying lymphocytic-variant hypereosinophilia. However, there is increasing recognition of small immunophenotypically unusual or abnormal populations of peripheral blood cells, particularly in older patients, which are of uncertain clinical significance, such as monoclonal B lymphocytosis and T-cell clonopathy. Therefore, it is important to integrate peripheral blood smear review findings with the clinical and other information before recommending flow cytometry. In addition, it is important to recognize situations where the results of peripheral blood smear review and flow cytometric immunophenotyping do not explain the clinical findings. © 2017 John Wiley & Sons Ltd.

  6. Increased oxidative stress and apoptosis in peripheral blood mononuclear cells of fructose-fed rats.

    PubMed

    Porto, Marcella L; Lírio, Layla M; Dias, Ananda T; Batista, Alan T; Campagnaro, Bianca P; Mill, José G; Meyrelles, Silvana S; Baldo, Marcelo P

    2015-12-01

    Measuring of oxidative stress in peripheral blood mononuclear cells is a suitable model of dietary induced systemic oxidative stress. Thus, we aimed to evaluate whether a chronic high fructose intake could induce oxidative damage in peripheral blood and bone marrow mononuclear cells of rats. Animals were randomly assigned to the following groups: Control group (standard rat chow and tap water n=8), and Fructose group (standard rat chow and a 10% fructose solution in the drinking water n=8). Reactive oxygen species and cytokines were measure using flow cytometry in peripheral blood and bone-marrow mononuclear cells. Apoptotic cell death and the advanced oxidation protein products (AOPP) were also determined. We observed a significant increase in ROS production in peripheral blood mononuclear cells of fructose group as compared to control rats. Apoptosis and the AOPP were higher in those animals underwent high fructose intake. Serum levels of IL-6 and IL-12 were also increased after 12 weeks of high fructose intake. We concluded that fructose intake leads to systemic oxidative stress and pro-inflammatory condition which affect peripheral blood mononuclear cells and bone-marrow mononuclear cells viability. Copyright © 2015 Elsevier B.V. All rights reserved.

  7. Transcriptional profiling of peripheral blood in pancreatic adenocarcinoma patients identifies diagnostic biomarkers.

    PubMed

    Caba, Octavio; Prados, Jose; Ortiz, Raúl; Jiménez-Luna, Cristina; Melguizo, Consolación; Alvarez, Pablo J; Delgado, Juan R; Irigoyen, Antonio; Rojas, Ignacio; Pérez-Florido, Javier; Torres, Carolina; Perales, Sonia; Linares, Ana; Aránega, Antonia

    2014-11-01

    Pancreatic ductal adenocarcinoma (PDAC) is a lethal malignancy associated with poor survival rates. Fast detection of PDAC appears to be the most relevant strategy to improve the long-term survival of patients. Our objective was to identify new markers in peripheral blood that differentiates between PDAC patients and healthy controls. Peripheral blood samples from PDAC patients (n = 18) and controls (n = 18) were analyzed by whole genome cDNA microarray hybridization. The most relevant genes were validated by quantitative real-time PCR (RT-qPCR) in the same set of samples. Finally, our gene prediction set was tested in a blinded set of new peripheral blood samples (n = 30). Microarray studies identified 87 genes differentially expressed in peripheral blood samples from PDAC patients. Four of these genes were selected for analysis by RT-qPCR, which confirmed the previously observed changes. In our blinded validation study, the combination of CLEC4D and IRAK3 predicted the diagnosis of PDAC with 93 % accuracy, with a sensitivity of 86 % and specificity of 100 %. Peripheral blood gene expression profiling is an useful tool for the diagnosis of PDAC. We present a validated four-gene predictor set (ANKRD22, CLEC4D, VNN1, and IRAK3) that may be useful in PDAC diagnosis.

  8. Modelling the relationship between peripheral blood pressure and blood volume pulses using linear and neural network system identification techniques.

    PubMed

    Allen, J; Murray, A

    1999-08-01

    The relationships between peripheral blood pressure and blood volume pulse waveforms can provide valuable physiological data about the peripheral vascular system, and are the subject of this study. Blood pressure and volume pulse waveforms were collected from 12 normal male subjects using non-invasive optical techniques, finger arterial blood pressure (BP, Finapres: Datex-Ohmeda) and photoelectric plethysmography (PPG) respectively, and captured to computer for three equal (1 min) measurement phases: baseline, hand raising and hand elevated. This simple physiological challenge was designed to induce a significant drop in peripheral blood pressure. A simple first order lag transfer function was chosen to study the relationship between blood pressure (system input) and blood volume pulse waveforms (system output), with parameters describing the dynamics (time constant, tau) and input-output gain (K). Tau and K were estimated for each subject using two different system identification techniques: a recursive parameter estimation algorithm which calculated tau and K from a linear auto-regressive with exogenous variable (ARX) model, and an artificial neural network which was trained to learn the non-linear process input-output relationships and then derive a linearized ARX model of the system. The identification techniques allowed the relationship between the blood pressure and blood volume pulses to be described simply, with the neural network technique providing a better model fit overall (p < 0.05, Wilcoxon). The median falls in tau following the hand raise challenge were 26% and 31% for the linear and neural network based techniques respectively (both p < 0.05, Wilcoxon). This preliminary study has shown that the time constant and gain parameters obtained using these techniques can provide physiological data for the clinical assessment of the peripheral circulation.

  9. Platelet and Red Blood Cell Utilization and Transfusion Independence in Umbilical Cord Blood and Allogeneic Peripheral Blood Hematopoietic Cell Transplants

    PubMed Central

    Solh, Melhem; Brunstein, Claudio; Morgan, Shanna; Weisdorf, Daniel

    2010-01-01

    Allogeneic hematopoietic cell transplantation (HCT) recipients have substantial transfusion requirements. Factors associated with increased transfusions and the extent of blood product use in umbilical cord blood (UCB) recipients are uncertain. We reviewed blood product use in 229 consecutive adult recipients of allogeneic HCT at the University of Minnesota: 147 with leukemia, 82 lymphoma or myeloma; 58% received unrelated UCB and 43% sibling donor peripheral blood stem cell (PBSC) grafts. Although neutrophil recovery was prompt (UCB median 17, range 2–45 days, and PBSC 14, range 3–34 days), only 135 of 229 (59% cumulative incidence, CI) achieved RBC independence and 157 (69%) achieved platelet independence by 6 months. Time to platelet independence was prolonged in UCB recipients (median UCB 41 vs. PBSC 14 days) and in patients who had received a prior transplant (median 48 vs. 32 days). Patients who received UCB grafts required more RBC through day 60 post HCT (mean UCB 7.8 (95% CI 6.7–8.9) vs. PBSC 5.2 (3.7–6.7) transfusions, p=0.04), and more platelet transfusions (mean 25.2 (95% CI 22.1–28.2) vs. 12.9 (9.4–16.4), p<0.01) compared to PBSC recipients. Patient receiving myeloablative (MA) conditioning required more RBC and platelet transfusions during the first 2 months post HCT compared to reduced intensity conditioning (RIC) (7.4 vs. 6.2, p=0.3 for RBC; 23.2 vs 17.5, p=0.07 for platelets). Despite prompt neutrophil engraftment, UCB recipients had delayed platelet recovery as well as more prolonged and costly blood product requirements. Enhanced approaches to accelerate multilineage engraftment could limit the transfusion-associated morbidity and costs accompanying UCB allotransplantation. PMID:20813199

  10. Candidemia Diagnosed from Peripheral Blood Smear: Case Report and Review of Literature 1954-2013.

    PubMed

    Hirai, Yuji; Asahata, Sayaka; Ainoda, Yusuke; Fujita, Takahiro; Miura, Hitomi; Hizuka, Naomi; Kikuchi, Ken

    2015-08-01

    Yeast with pseudohyphae or those that have been phagocytized by white blood cells are coincidentally found in peripheral blood smears. The clinical diagnostic value and outcome of candidaemia diagnosed from peripheral blood smears (CPBSs) are unclear. A 45-year-old man with diabetes and panhypopituitarism for 20 years received 10 mg of hydrocortisone and 100 μg of levothyroxine sodium hydrate daily. He has been admitted seven times because of adrenal failure triggered by infections and was admitted for pneumonia. On day 56, some budding yeast was found microscopically in a peripheral blood smear with May-Giemsa staining. Some of them were phagocytized by white blood cells. The two blood cultures yielded Candida parapsilosis. Despite antifungal treatment and removal of an intravenous catheter, on day 98 (42 days after the candidaemia diagnosis), the patient died. We analysed 36 cases including the present case. Almost all CPBS patients (96.5 %, n = 29) were using an intravenous catheter. The most frequently isolated species was C. parapsilosis (35.1 %), followed by C. albicans (29.7 %). The overall mortality rate was 53.6 % (n = 28). The time from the discovery of yeast-like pathogens using peripheral blood smears to death ranged from a few hours to 93 days (median 19 days). The present results suggest that intravenous catheter use and the underlying conditions of patients are responsible for CPBSs. The detection of yeast in peripheral blood smears suggests advanced infections with uncontrollable complications, which means a poor prognosis. Rapid detection methods besides blood culture are needed.

  11. Peripheral Blood Mitochondrial DNA Damage as a Potential Noninvasive Biomarker of Diabetic Retinopathy

    PubMed Central

    Mishra, Manish; Lillvis, John; Seyoum, Berhane; Kowluru, Renu A.

    2016-01-01

    Purpose In the development of diabetic retinopathy, retinal mitochondria become dysfunctional, and mitochondrial DNA (mtDNA) is damaged. Because retinopathy is a progressive disease, and circulating glucose levels are high in diabetes, our aim was to investigate if peripheral blood mtDNA damage can serve as a potential biomarker of diabetic retinopathy. Methods Peripheral blood mtDNA damage was investigated by extended-length PCR in rats and mice, diabetic for 10 to 12 months (streptozotocin-induced, type 1 model), and in 12- and 40-week-old Zucker diabetic fatty rats (ZDF, type 2). Mitochondrial copy number (in gDNA) and transcription (in cDNA) were quantified by qPCR. Similar parameters were measured in blood from diabetic patients with/without retinopathy. Results Peripheral blood from diabetic rodents had significantly increased mtDNA damage and decreased copy numbers and transcription. Lipoic acid administration in diabetic rats, or Sod2 overexpression or MMP-9 knockdown in mice, the therapies that prevent diabetic retinopathy, also ameliorated blood mtDNA damage and restored copy numbers and transcription. Although blood from 40-week-old ZDF rats had significant mtDNA damage, 12-week-old rats had normal mtDNA. Diabetic patients with retinopathy had increased blood mtDNA damage, and decreased transcription and copy numbers compared with diabetic patients without retinopathy and nondiabetic individuals. Conclusions Type 1 diabetic rodents with oxidative stress modulated by pharmacologic/genetic means, and type 2 animal model and patients with/without diabetic retinopathy, demonstrate a strong relation between peripheral blood mtDNA damage and diabetic retinopathy, and suggest the possibility of use of peripheral blood mtDNA as a noninvasive biomarker of diabetic retinopathy. PMID:27494345

  12. Enhanced external counterpulsation improves peripheral resistance artery blood flow in patients with coronary artery disease.

    PubMed

    Avery, Joseph C; Beck, Darren T; Casey, Darren P; Sardina, Paloma D; Braith, Randy W

    2014-03-01

    Enhanced external counterpulsation (EECP) increases coronary artery perfusion and improves endothelium-dependent vasodilation in peripheral muscular conduit arteries. It is unknown whether vasodilatory capacity is improved in the peripheral resistance vasculature. Here we provide novel evidence from the first randomized, sham-controlled study that EECP increases peak limb blood flow and improves endothelium-dependent vasodilation in both calf and forearm resistance arteries in patients with coronary artery disease.

  13. Pediatric blood sample collection from a pre-existing peripheral intravenous (PIV) catheter.

    PubMed

    Braniff, Heather; DeCarlo, Ann; Haskamp, Amy Corey; Broome, Marion E

    2014-01-01

    Aiming to minimize pain in a hospitalized child, the purpose of this observational study was to describe characteristics of blood samples collected from pre-existing peripheral intravenous (PIV) catheters in pediatric patients. One hundred and fifty blood samples were reviewed for number of unusable samples requiring a specimen to be re-drawn. Success of the blood draw and prevalence of the loss of the PIV following blood collection was also measured. Findings included one clotted specimen, success rate of 91.3%, and 1.3% of PIVs becoming non-functional after collection. Obtaining blood specimens from a pre-existing PIV should be considered in a pediatric patient.

  14. Isolation and two-step classification of normal white blood cells in peripheral blood smears.

    PubMed

    Ramesh, Nisha; Dangott, Bryan; Salama, Mohammed E; Tasdizen, Tolga

    2012-01-01

    An automated system for differential white blood cell (WBC) counting based on morphology can make manual differential leukocyte counts faster and less tedious for pathologists and laboratory professionals. We present an automated system for isolation and classification of WBCs in manually prepared, Wright stained, peripheral blood smears from whole slide images (WSI). A simple, classification scheme using color information and morphology is proposed. The performance of the algorithm was evaluated by comparing our proposed method with a hematopathologist's visual classification. The isolation algorithm was applied to 1938 subimages of WBCs, 1804 of them were accurately isolated. Then, as the first step of a two-step classification process, WBCs were broadly classified into cells with segmented nuclei and cells with nonsegmented nuclei. The nucleus shape is one of the key factors in deciding how to classify WBCs. Ambiguities associated with connected nuclear lobes are resolved by detecting maximum curvature points and partitioning them using geometric rules. The second step is to define a set of features using the information from the cytoplasm and nuclear regions to classify WBCs using linear discriminant analysis. This two-step classification approach stratifies normal WBC types accurately from a whole slide image. System evaluation is performed using a 10-fold cross-validation technique. Confusion matrix of the classifier is presented to evaluate the accuracy for each type of WBC detection. Experiments show that the two-step classification implemented achieves a 93.9% overall accuracy in the five subtype classification. Our methodology achieves a semiautomated system for the detection and classification of normal WBCs from scanned WSI. Further studies will be focused on detecting and segmenting abnormal WBCs, comparison of 20× and 40× data, and expanding the applications for bone marrow aspirates.

  15. Magnetohydrodynamic Voltage Recorder for Comparing Peripheral Blood Flow.

    PubMed

    Wu, Kevin J; Gregory, T Stan; Lastinger, Michael C; Murrow, Jonathan R; Tse, Zion Tsz Ho

    2017-06-22

    Blood flow is a clinical metric for monitoring of cardiovascular diseases but current measurements methods are costly or uncomfortable for patients. It was shown that the interaction of the magnetic field (B 0) during MRI and blood flow in the body, through the magnetohydrodynamic (MHD) effect, produce voltages (V MHD) observable through intra-MRI electrocardiography (ECG), which are correlated with regional blood flow. This study shows the reproducibility of V MHD outside the MRI and its application in a portable flow monitoring device. To recreate this interaction outside the MRI, a static neodymium magnet (0.4T) was placed in between two electrodes to induce the V MHD in a single lead ECG measurement. V MHD was extracted, and integrated over to obtain a stroke volume metric. A smartphone-enabled device utilizing this interaction was developed in order to create a more accessible method of obtaining blood flow measurements. The portable device displayed a <6% error compared to a commercial recorder, and was able to successfully record V MHD using the 0.4T magnet. Exercise stress testing showed a V MHD increase of 23% in healthy subjects, with an 81% increase in the athlete. The study demonstrates a new device utilizing MHD interactions with body circulation to obtain blood flow metrics.

  16. Axillary versus peripheral blood levels of sialic acid, ferritin, and CEA in patients with breast cancer.

    PubMed

    Monti, M; Catania, S; Locatelli, E; Gandini, R; Reggiani, A; Cunietti, E

    1990-12-01

    Serum levels of total sialic acid, carcinoembryonic antigen (CEA), ferritin, lactate dehydrogenase, and creatine phosphokinase were measured both in tumor drainage blood (axillary vein) and in peripheral blood obtained from 121 breast cancer patients during surgery. No significant differences between mean values in peripheral and tumor draining blood, between cancer patients and healthy controls, or between patients with or without axillary lymph node metastases were found for any of the markers. Both ferritin and CEA levels were higher in axillary and peripheral blood from patients with central breast cancer versus other sites but the difference was significant only for CEA (p less than 0.05). CEA levels were significantly higher (p less than 0.01) in patients with greater than 2 cm diameter carcinomas versus T1 stage patients in axillary but not in peripheral blood. When the cephalic vein was clamped before the axillary sample was taken, ferritin showed a significant increase (p less than 0.05). We conclude that measurement of sialic acid, CEA, and ferritin in axillary venous blood in breast cancer patients is not of clinical benefit, although further data are needed to clarify whether other advantages can be derived.

  17. Detection of microparticles of leukocytic origin in the peripheral blood in normal pregnancy and preeclampsia.

    PubMed

    Mikhailova, V A; Ovchinnikova, O M; Zainulina, M S; Sokolov, D I; Sel'kov, S A

    2014-10-01

    Microparticles are microvesicles forming during cell activation and as a result of apoptotic cell death. Normal pregnancy is associated with apoptosis induction in active immune system cells, present in the decidual tissue. Preeclampsia is associated with activation of the peripheral blood leukocytes and more intense apoptosis of the trophoblast cells. As a result, the number of microparticles in the peripheral blood is changing in normal gestation and in preeclampsia. The content of the leukocytic microparticles in the peripheral blood is evaluated in normal pregnancy and preeclampsia. The content of neutrophilic and monocytic microparticles is higher than normally in preeclampsia, this indicating activation of these cells. The number of microparticles formed by NK cells is low in preeclampsia, which can reflect the incompetence of immunological tolerance mechanisms under these conditions.

  18. Evaluation of genome-wide genotyping concordance between tumor tissues and peripheral blood.

    PubMed

    Shao, Wei; Ge, Yuqiu; Ma, Gaoxiang; Du, Mulong; Chu, Haiyan; Qiang, Fulin; Zhang, Zhengdong; Wang, Meilin

    2017-03-01

    Tumor tissues were potential resources in cancer susceptibility studies. To assess the genotyping concordance between tumor tissues and peripheral blood, we conducted this study in a large sample size and genome-wide scale. Genome-wide genotypes of human colon adenocarcinoma (COAD) retrieved from The Cancer Genome Atlas (TCGA) was analyzed. A total of 387 pairs of matched fresh frozen tumor tissues and peripheral blood samples passed the quality control processes. High concordant rate (94.85% with no-calls and 97.89% without no-calls) was found between tumor tissues and peripheral blood. The discordant rate raised with the increase of heterozygote rate, and the tendency was statistically significant. The total missing rate was 3.10%. We also verified 14 susceptibility SNPs and the average genotyping concordant rate was 97.42%. These findings suggest that majority of SNPs could be accurately genotyped using DNA isolated from tumor tissues. Copyright © 2017 Elsevier Inc. All rights reserved.

  19. Histochemical and morphometric studies of peripheral muscle in bovine progressive degenerative myeloencephalopathy of brown Swiss cattle.

    PubMed

    Oyster, R; Leipold, H W; Troyer, D; Cash, W; Johnson, D

    1992-06-01

    Histochemical and morphometric analysis of selected skeletal muscles was performed on 14 pure bred, Brown Swiss cattle. Nine cattle were clinically affected with bovine progressive degenerative myeloencephalopathy (BPDME) while five served as controls. Statistically significant trend differences were not observed for the parameters of mean cross sectional area, and mean fiber type percentages for types, I, IIA, and IIB fibers between affected and control test groups. In general, patterns of hypertrophy or atrophy, fiber type grouping, fiber type predominance, or fiber cross sectional profile alteration were not observed in the muscles examined from affected cattle. The findings suggest that BPDME, or weaver syndrome, is not a muscular dystrophy and that muscle pathology is not a primary part of the syndrome nor would muscle pathology be expected to contribute significantly to the clinical signs of the disease.

  20. Cationic pyridinium porphyrins appending different peripheral substituents: spectroscopic studies on their interactions with bovine serum albumin.

    PubMed

    Zhao, Ping; Huang, Jin-Wang; Ji, Liang-Nian

    2012-03-01

    The interaction of cationic pyridinium porphyrins appending methylpyridyl, hydroxyphenyl, propionoxyphenyl or carboxyphenyl group at meso-20-position of porphyrin core with bovine serum albumin (BSA), was studied by the combination of absorption spectroscopy, surface-enhanced Raman spectroscopy (SERS), circular dichroism (CD) spectroscopy, fluorescence spectroscopy and synchronous spectroscopy. The spectral monitoring results indicate that the studied compounds could bind with the BSA molecule and the calculated binding constants show that the tetracationic porphyrin has higher binding affinity than those tricationic ones. The interactions between porphyrins and BSA employ an electrostatic binding mechanism and there was only one binding site which located on the surface of the protein molecule. Copyright © 2011 Elsevier B.V. All rights reserved.

  1. A New Dietary Supplement Based on Bovine Blood: Recovering Brazilian Children from Malnutrition.

    PubMed

    Teodósio, N R; Perez, E P; Alves, A J; Dimenstein, W; Guedes, R C

    2000-01-01

    Two compounds ("Prothemol" and "Plasmel"), based on bovine blood as source of high quality-protein, were tested as supplement for malnourished children. Prothemol is a powder containing desiccated bovine red cells, with 23.32 g% protein and 18.8 mg% iron, without any limiting amino acid. Plasmel (a syrup) contains 44.7% bovine plasma, 54.3% saccharose and 1500 IU% retinol. Children, 32-60 month old, from a day-nursery service in Recife, Brazil, received Prothemol + Plasmel for 90 (n = 14) or 180 days (n = 8). When compared to age-matched control children (n = 12 and n = 6, respectively), they presented significantly higher increments in weight and height, and in some haematological parameters. Clinical signs associated to malnutrition (faces suggesting suffering or sadness; brightnessless eyes; apathy; reduced mobility; reduced communication with their classmates and adults) were found in 12 treated children (85.7%) and in 9 controls (75%). Recovery from these signs begun after 51 ± 20.6 and 103.5 ± 14.6 days, for the treated and control groups, respectively (P < 0.05) and occurred in 100% of the treated children and in 67% (6 of 9 children) in the controls. We suggest that Prothemol + Plasmel is an effective dietary supplement to help treating malnutrition in children, recovering them from clinical signs indicative of improving neural functions.

  2. [Phenotypic analysis of Th cells in colon and peripheral blood in patients with irritable bowel syndrome].

    PubMed

    Fu, Yu; Tong, Jing-Jing; Pan, Qi; Wang, Wen-Feng; Zou, Kai-Fang; Qian, Wei; Hou, Xiao-Hua

    2009-08-11

    OBJECTIVE; To analyze the change of Th1/Th2/Th17 in colonic mucosa and peripheral blood in diarrhea-predominant IBS (D-IBS) to uncover the underlying mechanism for the activation of mucosal immune system. Colonic biopsy specimens and peripheral blood were obtained from patients with D-IBS (n = 27) and controls (n = 16). Two different groups were classified on the basis of histological assessment of biopsy specimens from D-IBS patients. One group (14 of 27) had normal conventional histology (IBS), while another group (13 of 27) had nonspecific microscopic inflammation (IBS-A). Flow cytometric detection of intracellular IFN-gamma/IL-4/IL-17 cytokine production was employed to investigate Th1, Th2 and Th17 cells in colonic lamina propria and peripheral blood. Western blot was used to determine the expressions of IL-12, IL-4 and IL-17 in colonic mucosa. The levels of IL-12, IL-4 and IL-17 in peripheral blood were detected by ELISA. In colonic mucosa, the proportion of Th17 increased in IBS-A group as compared with controls [3.60 (4.05) vs 1.25 (3.70), P = 0.045], but not in IBS group. No difference could be observed in the frequencies of Th1 and Th2 in colonic mucosa and peripheral blood. The levels of IL-12, IL4 and IL-17 in IBS and IBS-A showed no difference in either colonic mucosa or peripheral blood. Subgroup of D-IBS showed abnormal conventional histology, implicating the activation of mucosal immune system in pathogenesis. The shift of Th1/Th2/Th17 balance in colonic mucosa showed the enhanced Th17 activity.

  3. The in vitro culture characteristics and requirements for expression of receptors for IgM on human peripheral blood lymphocytes.

    PubMed Central

    Golightly, M G; Golub, S H

    1980-01-01

    The in vitro culture characteristics and requirements for expression of receptors for the Fc portion of IgM (Fcmu) on peripheral blood lymphocytes was investigated. Significant numbers of Fcmu receptors cells are not detected until after 11 hr of in vitro culture. This was demonstrated not to be the result of shedding in vivo bound IgM thus unblocking the Fcmu receptors after culturing but rather an increased number of Fcmu receptors on cultures cells, presumable because of de novo synthesis. It was also shown that the Fcmu and Fc gamma receptors are not mutually exclusive in that a small population of PBL exhibit both Fcmu and Fc gamma receptors and that Fcmu negative cells may become Fcmu positive cells. Furthermore, it was demonstrated that fetal bovine serum and human blood group AB serum which are known to affect differentially various in vitro cell mediated functions also differentially affect Fcmu receptor expression and are capable of modulating the Fcmu receptor levels. These observations demonstrate the diverse and dynamic nature of the Fcmu receptor. PMID:7389194

  4. T-cell subsets in peripheral blood and tumors of patients treated with oncolytic adenoviruses.

    PubMed

    Kristian, Taipale; Ilkka, Liikanen; Juuso, Juhila; Aila, Karioja-Kallio; Minna, Oksanen; Riku, Turkki; Nina, Linder; Johan, Lundin; Ari, Ristimäki; Anna, Kanerva; Anniina, Koski; Timo, Joensuu; Markus, Vähä-Koskela; Akseli, Hemminki

    2015-05-01

    The quality of the antitumor immune response is decisive when developing new immunotherapies for cancer. Oncolytic adenoviruses cause a potent immunogenic stimulus and arming them with costimulatory molecules reshapes the immune response further. We evaluated peripheral blood T-cell subsets of 50 patients with refractory solid tumors undergoing treatment with oncolytic adenovirus. These data were compared to changes in antiviral and antitumor T cells, treatment efficacy, overall survival, and T-cell subsets in pre- and post-treatment tumor biopsies. Treatment caused a significant (P < 0.0001) shift in T-cell subsets in blood, characterized by a proportional increase of CD8(+) cells, and decrease of CD4(+) cells. Concomitant treatment with cyclophosphamide and temozolomide resulted in less CD4(+) decrease (P = 0.041) than cyclophosphamide only. Interestingly, we saw a correlation between T-cell changes in peripheral blood and the tumor site. This correlation was positive for CD8(+) and inverse for CD4(+) cells. These findings give insight to the interconnections between peripheral blood and tumor-infiltrating lymphocyte (TIL) populations regarding oncolytic virotherapy. In particular, our data suggest that induction of T-cell response is not sufficient for clinical response in the context of immunosuppressive tumors, and that peripheral blood T cells have a complicated and potentially misleading relationship with TILs.

  5. Cell survival kinetics in peripheral blood and bone marrow during total body irradiation for marrow transplantation

    SciTech Connect

    Shank, B.; Andreeff, M.; Li, D.

    1983-11-01

    Cell survival kinetics in both peripheral blood and in bone marrow have been studied over the time course of hyperfractionated total body irradiation (TBI) for bone marrow transplantation. Our unique TBI regimen allows the study of the in vivo radiation effect uncomplicated by prior cyclophosphamide, since this agent is given after TBI in our cytoreduction scheme. Peripheral blood cell concentrations were monitored with conventional laboratory cell counts and differentials. Absolute bone marrow cell concentrations were monitored by measuring cell concentrations in an aspirate sample and correcting for dilution with blood by a cell cycle kinetic method using cytofluorometry. For lymphocytes in peripheral blood in patients in remission, the effective D/sub 0/ ranged from 373 rad in 10 children less than or equal to 10 y old, to 536 rad in the four patients between 11 to 17 y old, while n = 1.0 in all groups. There was no trend observed according to age. Granulocytes had a much higher effective D/sub 0/, approximately 1000 rad in vivo. Absolute nucleated cell concentration in marrow dropped slowly initially, due to an increased lymphocyte concentration in marrow during a concurrent drop in lymphocyte concentration in peripheral blood, but eventually fell on the last day of TBI ranging from 7 to 44% of the initial marrow nucleated cell concentration. Marrow myeloid elements, however, dropped continuously throughout the course of TBI.

  6. Evaluating the presence of Toxoplasma gondii in peripheral blood of patients with diverse forms of uveitis.

    PubMed

    Belfort, Rubens N; Isenberg, Jordan; Fernandes, Bruno F; Di Cesare, Sebastian; Belfort, Rubens; Burnier, Miguel N

    2017-02-01

    The purpose of this study was to evaluate the presence of Toxoplasmosis gondii in samples of peripheral blood from patients with varying etiologies of uveitis. Whole blood from patients with different forms of uveitis was tested for the presence of T. gondii using real-time PCR targeting the well-characterized 529 bp fragment. Extracted DNA was both frozen. Thirty-one patients were included in the current study and grouped as follows: acute toxoplasmosis (n = 10); toxoplasmic retinal scars (n = 9); non-infectious etiologies of uveitis (n = 6); and IgG negative for toxoplasmosis (n = 6). In total, only two patients were shown to have circulating T. gondii in peripheral blood; both of these patients were IgG positive for toxoplasmosis, were receiving immunosuppressive therapy for autoimmune uveitis, and had no clinical features of toxoplasmosis. T. gondii was identified in peripheral blood of some immunosuppressed patients. No other patients, including those with acute toxoplasmosis, had circulating parasites in peripheral blood.

  7. A comparative study of bovine blood and milk neutrophil functions with luminol-dependent chemiluminescence.

    PubMed

    Mehrzad, J; Dosogne, H; Vangroenweghe, F; Burvenich, C

    2001-01-01

    In this study, a technique was developed for the chemiluminescence (CL) measurement of bovine milk polymorphonuclear leukocytes (PMN). In the first study, the effects of cell number and the concentration of phorbol-12-myristate-13-acetate (PMA), luminol, latex bead particles, dimethyl sulphoxide (DMSO) and gelatin on the luminol-dependent cellular CL (LDCL) response were assessed with healthy cows in different stages of lactation. In the second study, the LDCL and in vitro bactericidal activity of blood and milk PMN towards Staphylococcus aureus was investigated. In general, the CL activity of blood PMN was consistently higher than that of milk PMN. We found that (a) the optimal cell density in blood and milk cells for maximal LDCL response ranged from 1.5 x 10(6) to 5 x 10(6) cells/mL; (b) the optimal concentrations of PMA, latex beads and luminol for maximal LDCL response were 100-200 ng/ml, 500 particles/PMN and 0.1 mmol/L, respectively. Concentrations of DMSO of 0.5-1% (v/v) did not significantly affect the maximal CL response of PMN. Gelatin concentrations of 0.1 -0.5 mg/ml had no effect on the LDCL of PMN. In addition, the LDCL of PMN was significantly correlated with bactericidal activity towards S. aureus (r = 0.78, p < 0.001 for blood PMN and r = 0.66, p < 0.01 for milk PMN). Under the optimal experimental conditions for measurement of CL produced by bovine blood and milk PMN defined in this study, LDCL assay is an accurate and reproducible technique for the rapid quantification of PMN bactericidal activity in physiological and pathological conditions of high-yielding dairy cows. Copyright 2001 John Wiley & Sons, Ltd.

  8. Health status of birds fed diets containing three differently processed discarded vegetable-bovine blood-rumen content mixtures.

    PubMed

    Ekunseitan, D A; Balogun, O O; Sogunle, O M; Yusuf, A O; Ayoola, A A; Egbeyale, L T; Adeyemi, O A; Allison, I B; Iyanda, A I

    2013-04-01

    This study was conducted to determine the effect of feeding three differently processed mixtures on health status of broilers. A total of 1080 day-old Marshal broilers were fed; discarded vegetable-fresh bovine blood-fresh rumen digesta (P1), discarded vegetable-ensiled bovine blood-fresh rumen digesta (P2) and discarded vegetable-fresh bovine blood-ensiled rumen digesta (P3) at three levels of inclusion (0, 3 and 6%). Data on blood parameters was taken and were subjected to 3 x 3 factorial arrangements in a completely randomized design. Birds fed P1 had least values (p < 0.05) of serum glucose, total protein, globulin, uric acid and creatinine at starter phase. Birds fed diets containing 3 and 6% level of inclusion recorded the highest (p < 0.05) Packed cell volume, Haemoglobin, White blood cell and Red blood cell values. However, those fed at 0% level of inclusion recorded the highest albumin value. At finisher phase, birds fed P2 and P3 had the highest glucose, uric acid and creatinine values. 6% level of inclusion significantly (p < 0.05) increased the total protein and albumin values. Therefore, for enhanced performance and without comprising the health condition of birds; broiler chickens could be fed diets containing discarded vegetable-fresh bovine blood-ensiled rumen digesta (P3) up to 6% level of inclusion.

  9. DNA Methylation in Peripheral Blood: A Potential Biomarker for Cancer Molecular Epidemiology

    PubMed Central

    Li, Lian; Choi, Ji-Yeob; Lee, Kyoung-Mu; Sung, Hyuna; Park, Sue K.; Oze, Isao; Pan, Kai-Feng; You, Wei-Cheng; Chen, Ying-Xuan; Fang, Jing-Yuan; Matsuo, Keitaro; Kim, Woo Ho; Yuasa, Yasuhito; Kang, Daehee

    2012-01-01

    Aberrant DNA methylation is associated with cancer development and progression. There are several types of specimens from which DNA methylation pattern can be measured and evaluated as an indicator of disease status (from normal biological process to pathologic condition) and even of pharmacologic response to therapy. Blood-based specimens such as cell-free circulating nucleic acid and DNA extracted from leukocytes in peripheral blood may be a potential source of noninvasive cancer biomarkers. In this article, we describe the characteristics of blood-based DNA methylation from different biological sources, detection methods, and the factors affecting DNA methylation. We provide a comprehensive literature review of blood-based DNA methylation as a cancer biomarker and focus on the study of DNA methylation using peripheral blood leukocytes. Although DNA methylation patterns measured in peripheral blood have great potential to be useful and informative biomarkers of cancer risk and prognosis, large systematic and unbiased prospective studies that consider biological plausibility and data analysis issues will be needed in order to develop a clinically feasible blood-based assay. PMID:22863985

  10. Presence of Gumprecht shadows (smudge cells) in bovine leukemia virus-positive cattle.

    PubMed

    Panei, Carlos Javier; Larsen, Alejandra; González, Ester Teresa; Echeverría, María Gabriela

    2013-11-01

    Enzootic Bovine Leukosis is a chronic disease caused by the bovine leukemia virus (BLV). Smudge cells, also known as Gumprecht shadows, are not simple artifacts of slide preparation, but ragged lymphoid cells found mainly in peripheral blood smears from human patients with chronic lymphocytic leukemia. In this study, we report the presence of Gumprecht shadows in peripheral blood from BLV-positive cattle. Copyright © 2013 Elsevier B.V. All rights reserved.

  11. The expression, purification, crystallization and preliminary X-ray analysis of a subcomplex of the peripheral stalk of ATP synthase from bovine mitochondria

    SciTech Connect

    Silvester, Jocelyn A.; Kane Dickson, Veronica; Runswick, Michael J.; Leslie, Andrew G. W.; Walker, John E.

    2006-06-01

    A recombinant subcomplex of the peripheral stalk or stator domain of the ATP synthase from bovine mitochondria has been crystallized and a native data set has been collected to 2.8 Å resolution. A subcomplex of the peripheral stalk or stator domain of the ATP synthase from bovine mitochondria has been expressed to high levels in a soluble form in Escherichia coli. The subcomplex consists of residues 79–184 of subunit b, residues 1–124 of subunit d and the entire F{sub 6} subunit (76 residues). It has been purified and crystallized by vapour diffusion. The morphology and diffraction properties of the crystals of the subcomplex were improved by the presence of thioxane or 4-methylpyridine in the crystallization liquor. With a synchrotron-radiation source, these crystals diffracted to 2.8 Å resolution. They belong to the monoclinic space group P2{sub 1}.

  12. Role of peripheral blood mononuclear cell transportation from mother to baby in HBV intrauterine infection.

    PubMed

    Shao, Qingliang; Zhao, Xiaxia; Yao Li, M D

    2013-12-01

    We aimed to investigate the role of peripheral blood mononuclear cell transportation from mother to baby in hepatitis B virus (HBV) intrauterine infection. Thirty HBsAg-positive pregnant women in the second trimester and their aborted fetuses were included in this study. Enzyme-linked-immunosorbent-assay was utilized to detect HBsAg in the peripheral blood of pregnant women and the femoral vein blood of their aborted fetuses. HBV-DNA in serum and peripheral blood mononuclear cells (PBMC) and GSTM1 alleles of pregnant women and their aborted fetuses were detected by nested polymerase chain reaction (PCR) and seminested PCR, respectively. We also examined the location of placenta HBsAg and HBcAb using immunohistochemical staining. The expression of placenta HBV-DNA was detected by in situ hybridization. For the 30 aborted fetuses, the HBV intrauterine infection rate was 43.33%. The HBV-positive rates of HBsAg in peripheral blood, serum, and PBMC were 10% (3/30), 23.33% (7/30), and 33.33% (10/30), respectively. Maternal-fetal PBMC transport was significantly positively correlated with fetal PBMC HBV-DNA (P = 0.004). Meanwhile, the rates of HBV infection gradually decreased from the maternal side to the fetus side of placenta (decidual cells > trophoblastic cells > villous mesenchymal cells > villous capillary endothelial cells). However, no significant correlation between placenta HBV infection and HBV intrauterine infection was observed (P = 0.410). HBV intrauterine infection was primarily due to peripheral blood mononuclear cell maternal-fetal transportation in the second trimester in pregnant women.

  13. Device to determine the level of peripheral blood circulation and saturation

    NASA Astrophysics Data System (ADS)

    Kozlovska, Tetyana I.; Sander, Sergii V.; Zlepko, Sergii M.; Vasilenko, Valentina B.; Pavlov, Volodymyr S.; Dumenko, Victoria P.; Klapouschak, Andrii Yu.; Maciejewski, Marcin; DzierŻak, RóŻa; Surtel, Wojciech

    2016-09-01

    The paper evaluated the diagnostic value of laser photoplethysmography when examining patients with chronic lower limb ischemia. A statistical analysis of the research results was made, and diagrams of relationship between the degrees of ischemia and blood flow are presented. Development of the device to determine the level of peripheral blood circulation and saturation was presented. Also additional accessories in the form of optical fibers for different applications were suggested.

  14. The transcriptional landscape of age in human peripheral blood.

    PubMed

    Peters, Marjolein J; Joehanes, Roby; Pilling, Luke C; Schurmann, Claudia; Conneely, Karen N; Powell, Joseph; Reinmaa, Eva; Sutphin, George L; Zhernakova, Alexandra; Schramm, Katharina; Wilson, Yana A; Kobes, Sayuko; Tukiainen, Taru; Ramos, Yolande F; Göring, Harald H H; Fornage, Myriam; Liu, Yongmei; Gharib, Sina A; Stranger, Barbara E; De Jager, Philip L; Aviv, Abraham; Levy, Daniel; Murabito, Joanne M; Munson, Peter J; Huan, Tianxiao; Hofman, Albert; Uitterlinden, André G; Rivadeneira, Fernando; van Rooij, Jeroen; Stolk, Lisette; Broer, Linda; Verbiest, Michael M P J; Jhamai, Mila; Arp, Pascal; Metspalu, Andres; Tserel, Liina; Milani, Lili; Samani, Nilesh J; Peterson, Pärt; Kasela, Silva; Codd, Veryan; Peters, Annette; Ward-Caviness, Cavin K; Herder, Christian; Waldenberger, Melanie; Roden, Michael; Singmann, Paula; Zeilinger, Sonja; Illig, Thomas; Homuth, Georg; Grabe, Hans-Jörgen; Völzke, Henry; Steil, Leif; Kocher, Thomas; Murray, Anna; Melzer, David; Yaghootkar, Hanieh; Bandinelli, Stefania; Moses, Eric K; Kent, Jack W; Curran, Joanne E; Johnson, Matthew P; Williams-Blangero, Sarah; Westra, Harm-Jan; McRae, Allan F; Smith, Jennifer A; Kardia, Sharon L R; Hovatta, Iiris; Perola, Markus; Ripatti, Samuli; Salomaa, Veikko; Henders, Anjali K; Martin, Nicholas G; Smith, Alicia K; Mehta, Divya; Binder, Elisabeth B; Nylocks, K Maria; Kennedy, Elizabeth M; Klengel, Torsten; Ding, Jingzhong; Suchy-Dicey, Astrid M; Enquobahrie, Daniel A; Brody, Jennifer; Rotter, Jerome I; Chen, Yii-Der I; Houwing-Duistermaat, Jeanine; Kloppenburg, Margreet; Slagboom, P Eline; Helmer, Quinta; den Hollander, Wouter; Bean, Shannon; Raj, Towfique; Bakhshi, Noman; Wang, Qiao Ping; Oyston, Lisa J; Psaty, Bruce M; Tracy, Russell P; Montgomery, Grant W; Turner, Stephen T; Blangero, John; Meulenbelt, Ingrid; Ressler, Kerry J; Yang, Jian; Franke, Lude; Kettunen, Johannes; Visscher, Peter M; Neely, G Gregory; Korstanje, Ron; Hanson, Robert L; Prokisch, Holger; Ferrucci, Luigi; Esko, Tonu; Teumer, Alexander; van Meurs, Joyce B J; Johnson, Andrew D

    2015-10-22

    Disease incidences increase with age, but the molecular characteristics of ageing that lead to increased disease susceptibility remain inadequately understood. Here we perform a whole-blood gene expression meta-analysis in 14,983 individuals of European ancestry (including replication) and identify 1,497 genes that are differentially expressed with chronological age. The age-associated genes do not harbor more age-associated CpG-methylation sites than other genes, but are instead enriched for the presence of potentially functional CpG-methylation sites in enhancer and insulator regions that associate with both chronological age and gene expression levels. We further used the gene expression profiles to calculate the 'transcriptomic age' of an individual, and show that differences between transcriptomic age and chronological age are associated with biological features linked to ageing, such as blood pressure, cholesterol levels, fasting glucose, and body mass index. The transcriptomic prediction model adds biological relevance and complements existing epigenetic prediction models, and can be used by others to calculate transcriptomic age in external cohorts.

  15. Peripheral and Central Effects of Melatonin on Blood Pressure Regulation

    PubMed Central

    Pechanova, Olga; Paulis, Ludovit; Simko, Fedor

    2014-01-01

    The pineal hormone, melatonin (N-acetyl-5-methoxytryptamine), shows potent receptor-dependent and -independent actions, which participate in blood pressure regulation. The antihypertensive effect of melatonin was demonstrated in experimental and clinical hypertension. Receptor-dependent effects are mediated predominantly through MT1 and MT2 G-protein coupled receptors. The pleiotropic receptor-independent effects of melatonin with a possible impact on blood pressure involve the reactive oxygen species (ROS) scavenging nature, activation and over-expression of several antioxidant enzymes or their protection from oxidative damage and the ability to increase the efficiency of the mitochondrial electron transport chain. Besides the interaction with the vascular system, this indolamine may exert part of its antihypertensive action through its interaction with the central nervous system (CNS). The imbalance between the sympathetic and parasympathetic vegetative system is an important pathophysiological disorder and therapeutic target in hypertension. Melatonin is protective in CNS on several different levels: It reduces free radical burden, improves endothelial dysfunction, reduces inflammation and shifts the balance between the sympathetic and parasympathetic system in favor of the parasympathetic system. The increased level of serum melatonin observed in some types of hypertension may be a counter-regulatory adaptive mechanism against the sympathetic overstimulation. Since melatonin acts favorably on different levels of hypertension, including organ protection and with minimal side effects, it could become regularly involved in the struggle against this widespread cardiovascular pathology. PMID:25299692

  16. The transcriptional landscape of age in human peripheral blood

    PubMed Central

    Peters, Marjolein J.; Joehanes, Roby; Pilling, Luke C.; Schurmann, Claudia; Conneely, Karen N.; Powell, Joseph; Reinmaa, Eva; Sutphin, George L.; Zhernakova, Alexandra; Schramm, Katharina; Wilson, Yana A.; Kobes, Sayuko; Tukiainen, Taru; Nalls, Michael A.; Hernandez, Dena G.; Cookson, Mark R.; Gibbs, Raphael J.; Hardy, John; Ramasamy, Adaikalavan; Zonderman, Alan B.; Dillman, Allissa; Traynor, Bryan; Smith, Colin; Longo, Dan L.; Trabzuni, Daniah; Troncoso, Juan; van der Brug, Marcel; Weale, Michael E.; O'Brien, Richard; Johnson, Robert; Walker, Robert; Zielke, Ronald H.; Arepalli, Sampath; Ryten, Mina; Singleton, Andrew B.; Ramos, Yolande F.; Göring, Harald H. H.; Fornage, Myriam; Liu, Yongmei; Gharib, Sina A.; Stranger, Barbara E.; De Jager, Philip L.; Aviv, Abraham; Levy, Daniel; Murabito, Joanne M.; Munson, Peter J.; Huan, Tianxiao; Hofman, Albert; Uitterlinden, André G.; Rivadeneira, Fernando; van Rooij, Jeroen; Stolk, Lisette; Broer, Linda; Verbiest, Michael M. P. J.; Jhamai, Mila; Arp, Pascal; Metspalu, Andres; Tserel, Liina; Milani, Lili; Samani, Nilesh J.; Peterson, Pärt; Kasela, Silva; Codd, Veryan; Peters, Annette; Ward-Caviness, Cavin K.; Herder, Christian; Waldenberger, Melanie; Roden, Michael; Singmann, Paula; Zeilinger, Sonja; Illig, Thomas; Homuth, Georg; Grabe, Hans-Jörgen; Völzke, Henry; Steil, Leif; Kocher, Thomas; Murray, Anna; Melzer, David; Yaghootkar, Hanieh; Bandinelli, Stefania; Moses, Eric K.; Kent, Jack W.; Curran, Joanne E.; Johnson, Matthew P.; Williams-Blangero, Sarah; Westra, Harm-Jan; McRae, Allan F.; Smith, Jennifer A.; Kardia, Sharon L. R.; Hovatta, Iiris; Perola, Markus; Ripatti, Samuli; Salomaa, Veikko; Henders, Anjali K.; Martin, Nicholas G.; Smith, Alicia K.; Mehta, Divya; Binder, Elisabeth B.; Nylocks, K Maria; Kennedy, Elizabeth M.; Klengel, Torsten; Ding, Jingzhong; Suchy-Dicey, Astrid M.; Enquobahrie, Daniel A.; Brody, Jennifer; Rotter, Jerome I.; Chen, Yii-Der I.; Houwing-Duistermaat, Jeanine; Kloppenburg, Margreet; Slagboom, P. Eline; Helmer, Quinta; den Hollander, Wouter; Bean, Shannon; Raj, Towfique; Bakhshi, Noman; Wang, Qiao Ping; Oyston, Lisa J.; Psaty, Bruce M.; Tracy, Russell P.; Montgomery, Grant W.; Turner, Stephen T.; Blangero, John; Meulenbelt, Ingrid; Ressler, Kerry J.; Yang, Jian; Franke, Lude; Kettunen, Johannes; Visscher, Peter M.; Neely, G. Gregory; Korstanje, Ron; Hanson, Robert L.; Prokisch, Holger; Ferrucci, Luigi; Esko, Tonu; Teumer, Alexander; van Meurs, Joyce B. J.; Johnson, Andrew D.

    2015-01-01

    Disease incidences increase with age, but the molecular characteristics of ageing that lead to increased disease susceptibility remain inadequately understood. Here we perform a whole-blood gene expression meta-analysis in 14,983 individuals of European ancestry (including replication) and identify 1,497 genes that are differentially expressed with chronological age. The age-associated genes do not harbor more age-associated CpG-methylation sites than other genes, but are instead enriched for the presence of potentially functional CpG-methylation sites in enhancer and insulator regions that associate with both chronological age and gene expression levels. We further used the gene expression profiles to calculate the ‘transcriptomic age' of an individual, and show that differences between transcriptomic age and chronological age are associated with biological features linked to ageing, such as blood pressure, cholesterol levels, fasting glucose, and body mass index. The transcriptomic prediction model adds biological relevance and complements existing epigenetic prediction models, and can be used by others to calculate transcriptomic age in external cohorts. PMID:26490707

  17. [The association between cellular morphological changes in peripheral blood smear and complications in pediatric burn cases].

    PubMed

    Bozkurt, Mehmet; Kuvat, Samet Vasfi; Kapı, Emin; Karakol, Perçin; Ozel, Abdulkadir; Baykan, Halit

    2011-03-01

    Mortality and morbidity in burn cases can be reduced with early diagnosis. Many markers are used for early diagnosis of burn complications like sepsis. In this current study, the relationship between numerical/morphologic granulocyte abnormalities and complications was investigated in pediatric burns. It was aimed to introduce histopathologic marker(s) for burn-related complications. Thirty-two pediatric burn cases hospitalized between December 2006 and December 2009 were included in the study. A total of 192 complete blood count and peripheral blood smear results were analyzed comparatively. Findings were used to identify any correlation among white blood cell count and peripheral blood smear changes (the appearance of immature granular cells, toxic granulation, purple granules and Döhle bodies) and complications such as bacteriemia, sepsis, wound infections, severe anemia, and graft failure. White blood cell count changes and the appearance of immature granular cells were not suitable for use as a diagnostic marker for complications. Nevertheless, there was a statistically significant correlation between the appearance of toxic granulation, purple granules and Döhle bodies and subsequent complications (p: <0.0001, 0.041, 0.001, respectively). Toxic granulation, purple granules and Döhle bodies appear to be helpful in predicting burn-related complications. Therefore, peripheral blood smear is a suitable test for predicting future complications.

  18. Longitudinal peripheral blood transcriptional analysis of a patient with severe Ebola virus disease.

    PubMed

    Kash, John C; Walters, Kathie-Anne; Kindrachuk, Jason; Baxter, David; Scherler, Kelsey; Janosko, Krisztina B; Adams, Rick D; Herbert, Andrew S; James, Rebekah M; Stonier, Spencer W; Memoli, Matthew J; Dye, John M; Davey, Richard T; Chertow, Daniel S; Taubenberger, Jeffery K

    2017-04-12

    The 2013-2015 outbreak of Ebola virus disease in Guinea, Liberia, and Sierra Leone was unprecedented in the number of documented cases, but there have been few published reports on immune responses in clinical cases and their relationships with the course of illness and severity of Ebola virus disease. Symptoms of Ebola virus disease can include severe headache, myalgia, asthenia, fever, fatigue, diarrhea, vomiting, abdominal pain, and hemorrhage. Although experimental treatments are in development, there are no current U.S. Food and Drug Administration-approved vaccines or therapies. We report a detailed study of host gene expression as measured by microarray in daily peripheral blood samples collected from a patient with severe Ebola virus disease. This individual was provided with supportive care without experimental therapies at the National Institutes of Health Clinical Center from before onset of critical illness to recovery. Pearson analysis of daily gene expression signatures revealed marked gene expression changes in peripheral blood leukocytes that correlated with changes in serum and peripheral blood leukocytes, viral load, antibody responses, coagulopathy, multiple organ dysfunction, and then recovery. This study revealed marked shifts in immune and antiviral responses that preceded changes in medical condition, indicating that clearance of replicating Ebola virus from peripheral blood leukocytes is likely important for systemic viral clearance. Copyright © 2017, American Association for the Advancement of Science.

  19. Effects of oral eicosapentaenoic acid versus docosahexaenoic acid on human peripheral blood mononuclear cell gene expression

    USDA-ARS?s Scientific Manuscript database

    Objective: Eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) have beneficial effects on inflammation and cardiovascular disease (CVD). Our aim was to assess the effect of a six-week supplementation with either olive oil, EPA, or DHA on gene expression in peripheral blood mononuclear cells (...

  20. Induction of tumor necrosis factor in human peripheral-blood mononuclear cells by proteolytic enzymes.

    PubMed

    Desser, L; Rehberger, A

    1990-01-01

    We could demonstrate that polyenzyme preparations as well as bromelain and papain stimulate the production of tumor necrosis factor-alpha in human peripheral-blood mononuclear cell cultures in a time-dependent manner. We give evidence that immunomodulation and especially the release of cytokines may contribute to the therapeutic effect of these preparations.

  1. Global DNA hypomethylation in peripheral blood mononuclear cells as a biomarker of cancer risk

    USDA-ARS?s Scientific Manuscript database

    Global DNA hypomethylation is an early molecular event in carcinogenesis. Whether methylation measured in peripheral blood mononuclear cells (PBMCs) DNA is a clinically reliable biomarker for early detection or cancer risk assessment is to be established. From an original sample-set of 753 male and...

  2. An improved technique for peripheral blood stem cell collection in small patients.

    PubMed

    Landolfo, A; Angioni, A; Deb, G; Jenkner, A; Orlando, P; Donfrancesco, A; Balloni, P

    1991-03-01

    Peripheral Blood Stem Cell collection in adults is a consolidated method for autografting. The same procedure is less easily performed in pediatric patients due to low weight, vascular access problems and anticoagulant side effects. Great efforts have been made to overcome technical difficulties and make apheresis in children a reliable and safe procedure.

  3. M2b monocytes predominated in peripheral blood of severely burned patients.

    PubMed

    Kobayashi, Makiko; Jeschke, Marc G; Shigematsu, Kenji; Asai, Akira; Yoshida, Shohei; Herndon, David N; Suzuki, Fujio

    2010-12-15

    Severely burned patients were shown to be carriers of M2 monocytes, and all of the monocytes isolated from peripheral blood of severely burned patients (19 of 19 patients) were demonstrated as M2b monocytes (IL-12(-)IL-10(+)CCL1(+) monocytes). Low levels of M2a (IL-12(-)IL-10(+)CCL17(+) monocytes) and M2c monocytes (IL-12(-)IL-10(+)CXCL13(+) monocytes) were demonstrated in peripheral blood of severely burned patients (M2a, 2 of 19 patients; M2c, 5 of 19 patients). M2b, M2a, and M2c monocytes were not detected in peripheral blood of healthy donors. However, M2b monocytes appeared when healthy donor monocytes were cultured in media supplemented with burn patient serum (15%). CCL2 was detected in sera of all burn patients, and M2b monocytes were not generated from healthy donor monocytes cultured with media containing 15% burn patient sera that were previously treated with anti-CCL2 mAb. In addition, M2b monocytes were generated from healthy donor monocytes in cultures supplemented with rCCL2. These results indicate that M2b monocytes are predominant in peripheral blood of severely burned patients who are carriers of CCL2 that functions to stimulate monocyte conversion from resident monocytes to M2b monocytes.

  4. Noninvasive prediction of prostatic DNA damage by oxidative stress challenge of peripheral blood lymphocytes

    USDA-ARS?s Scientific Manuscript database

    To move closer to the goal of individualized risk prediction for prostate cancer, we used an in vivo canine model to evaluate whether genetic instability, expressed as the susceptibility of peripheral blood lymphocytes (PBLs) to oxidative stress-induced DNA damage, could identify those individuals w...

  5. Differential Infectivity and Division of Toxoplasma gondii in Human Peripheral Blood Leukocytes

    PubMed Central

    Channon, Jacqueline Y.; Seguin, Rosanne M.; Kasper, Lloyd H.

    2000-01-01

    When tachyzoites were incubated with human peripheral blood leukocytes in vitro, more monocytes and dendritic cells than neutrophils or lymphocytes were infected. Although tachyzoites were able to divide in each of these cell types, monocytes and dendritic cells were more permissive to rapid tachyzoite division than neutrophils or lymphocytes. PMID:10899898

  6. Luminescent analysis of lymphocytes of peripheral blood in definition of organism's sensitivity to specific antigene

    NASA Astrophysics Data System (ADS)

    Yusupova, Lira B.

    1994-01-01

    Possibility for definition of organism's sensitivity to specific allergen by means of luminescent analysis of peripheral blood lymphocytes was shown. The positive correlation dependence between luminescence intensity increase at 640 nm of acridine orange colored lymphocytes after simulation by specific allergene in vitro and the serum antiallergene antibodies level was detected.

  7. Cytogenetic biomonitoring of peripheral blood and oral mucosa cells from car painters.

    PubMed

    Pereira da Silva, Victor Hugo; Gomes de Moura, Carolina Foot; Spadari-Bratfisch, Regina Célia; Araki Ribeiro, Daniel

    2012-09-01

    The aim of the present study was to comparatively evaluate genomic damage and cellular death in exfoliated oral mucosa cells and peripheral blood from car painters. A total of 24 car painters and 19 healthy controls (non-exposed individuals) were included in this setting. Individuals had epithelial cells from cheek mucosa (left and right side) mechanically exfoliated, placed in fixative and dropped in clean slides which were checked for the specific nuclear phenotypes. A total of 5 μL from peripheral blood was collected for the single cell gel (comet) assay. The results pointed out statistically significant differences (p < 0.05) of micronucleated oral mucosa cells from car painters. In addition, DNA damage was detected in peripheral blood cells by single cell gel (comet) assay. Nevertheless, exposure to car paints did not cause increases other nuclear alterations closely related to cytotoxicity such as karrhyorexis, pyknosis and karyolysis in buccal mucosa cells. In summary, the results of the present study suggest that car painters comprise a high risk group since paints can induce genotoxic and mutagenic effects in peripheral blood and oral mucosa cells, respectively.

  8. Glutamate decreases the secretion of IL-10 by peripheral blood lymphocytes in persons with autoimmune thyroiditis.

    PubMed

    Kvaratskhelia, E; Dabrundashvili, N; Gagua, M; Maisuradze, E; Mikeladze, D

    2008-11-01

    Human T lymphocytes expose ionotropic and metabotropic glutamate receptors, which control immune responses, cell activation, maturation, and death. Several cytokines release during inflammation which identification may have important physiological and clinical implications. Main biological function of IL-10 is limitation and termination of inflammatory responses and the regulation of differentiation and proliferation of several immune cells. Various inflammatory molecules regulated the secretion of IL-8 and IL-10, but the action of glutamate on the biosynthesis of cytokines is unknown. We have found that in peripheral blood lymphocytes glutamate at the concentrations within normal plasma levels (1 x 10(-5) M), as well as at lower concentration (0.3 x 10(-6) M) changes the secretion of immunosuppressive cytokine IL-10, whereas synthesis of proinflammatory chemokine, IL-8 did not changed significantly. Moreover, our results have shown that peripheral blood lymphocytes from patients with autoimmune thyroiditis release less IL-10 at both concentration of glutamate than peripheral blood lymphocytes from healthy persons. These data suggest that glutamate decrease the secretion of IL-10 by peripheral blood lymphocytes, especially in patients with autoimmune thyroiditis that may be responsible for prolongation of inflammation.

  9. [Production of interleukin-2 by peripheral blood lymphocytes from patients with soft tissue sarcomas].

    PubMed

    Berezhnaia, N M; Goretskiĭ, B A; Konovalenko, V F; Palivets, A Iu; Tolstopiatov, B A

    1987-01-01

    Interleukin-2 (IL-2) production of phytohemagglutinin-stimulated peripheral blood lymphocytes (PBL) was studied in 9 healthy subjects and 19 patients with soft tissue sarcomas. Mean IL-2 production by PBL in 19 patients was significantly diminished as compared with the control. Surgery leads to an increase of IL-2 production, however, the levels observed in the control do not restore completely.

  10. Mobilization of hematopoietic stem cells into the peripheral blood.

    PubMed

    Damon, Lloyd E; Damon, Lauren E

    2009-12-01

    Hematopoietic stem cells can be mobilized out of the bone marrow into the blood for the reconstitution of hematopoiesis following high-dose therapy. Methods to improve mobilization efficiency and yields are rapidly emerging. Traditional methods include chemotherapy with or without myeloid growth factors. Plerixafor, a novel agent that disrupts the CXCR4-CXCL12 bond, the primary hematopoietic stem cell anchor in the bone marrow, has recently been US FDA-approved for mobilizing hematopoietic stem cells in patients with non-Hodgkin lymphoma and multiple myeloma. Plerixafor and myeloid growth factors as single agents appear safe to use in family or volunteer hematopoietic stem cells donors. Plerixafor mobilizes leukemic stem cells and is not approved for use in patients with acute leukemia. Patients failing to mobilize adequate hematopoietic stem cells with myeloid growth factors can often be successfully mobilized with chemotherapy plus myeloid growth factors or with plerixafor and granulocyte colony-stimulating factor.

  11. Evaluation of Four Veterinary Hematology Analyzers for Bovine and Ovine Blood Counts for In Vitro Testing of Medical Devices.

    PubMed

    Pieper, Ina Laura; Friedmann, Yasmin; Jones, Alyssa; Thornton, Catherine

    2016-04-18

    Small affordable automated hematology analyzers that produce rapid and accurate complete blood cell counts are a valuable tool to researchers developing blood-handling medical devices, such as ventricular assist devices, for in vitro safety assessments. In such studies, it is common to use the blood of large animals such as cattle and sheep. However, the commercially available instruments have not been evaluated for their ability to measure the blood counts of these animals. In this study, we compare, for the first time, four veterinary analyzers for blood counts on bovine and ovine blood samples. We look at ease of use, repeatability and agreement with a view to inform researchers of the benefits of these instruments in routine measurement of ovine and bovine bloods during in vitro testing. Complete blood cell counts and a three-part differential (granulocytes, monocytes, and lymphocytes) were measured by each of the instruments, and the results compared to those obtained from two additional analyzers used in a reference laboratory. Repeatability and agreement were evaluated using the Bland-Altman method; bias and 95% limits of agreement between the instruments, and between the instruments and two reference instruments, were used to evaluate instrument performance. In summary, there are advantages and disadvantages with all instruments. Of the four instruments tested, the repeatability and agreement was fairly similar for all instruments except one instrument which cannot be recommended for bovine or ovine blood counts.

  12. Evaluation of Peripheral Blood Circulation Disorder in Scleroderma Patients Using an Optical Sensor with a Pressurization Mechanism

    PubMed Central

    Yamakoshi, Yoshiki

    2016-01-01

    Blood circulation function of peripheral blood vessels in skin dermis was evaluated employing an optical sensor with a pressurization mechanism using the blood outflow and reflow characteristics. The device contains a light source and an optical sensor. When applied to the skin surface, it first exerts the primary pressure (higher than the systolic blood pressure), causing an outflow of blood from the dermal peripheral blood vessels. After two heartbeats, the pressure is lowered (secondary pressure) and blood reflows into the peripheral blood vessels. Hemoglobin concentration, which changes during blood outflow and reflow, is derived from the received light intensity using the Beer–Lambert law. This method was evaluated in 26 healthy female volunteers and 26 female scleroderma patients. In order to evaluate the blood circulation function of the peripheral blood vessels of scleroderma patients, pressurization sequence which consists of primary pressure followed by secondary pressure was adopted. Blood reflow during the first heartbeat period after applying the secondary pressure of 40mmHg was (mean±SD) 0.059±0.05%mm for scleroderma patients and 0.173±0.104%mm for healthy volunteers. Blood reflow was significantly lower in scleroderma patients than in healthy volunteers (p<0.05). This result indicates that the information necessary for assessing blood circulation disorder of peripheral blood vessels in scleroderma patients is objectively obtained by the proposed method. PMID:27479094

  13. Evaluation of Peripheral Blood Circulation Disorder in Scleroderma Patients Using an Optical Sensor with a Pressurization Mechanism.

    PubMed

    Yamakoshi, Yoshiki; Motegi, Sei-Ichiro; Ishikawa, Osamu

    2016-01-01

    Blood circulation function of peripheral blood vessels in skin dermis was evaluated employing an optical sensor with a pressurization mechanism using the blood outflow and reflow characteristics. The device contains a light source and an optical sensor. When applied to the skin surface, it first exerts the primary pressure (higher than the systolic blood pressure), causing an outflow of blood from the dermal peripheral blood vessels. After two heartbeats, the pressure is lowered (secondary pressure) and blood reflows into the peripheral blood vessels. Hemoglobin concentration, which changes during blood outflow and reflow, is derived from the received light intensity using the Beer-Lambert law. This method was evaluated in 26 healthy female volunteers and 26 female scleroderma patients. In order to evaluate the blood circulation function of the peripheral blood vessels of scleroderma patients, pressurization sequence which consists of primary pressure followed by secondary pressure was adopted. Blood reflow during the first heartbeat period after applying the secondary pressure of 40mmHg was (mean±SD) 0.059±0.05%mm for scleroderma patients and 0.173±0.104%mm for healthy volunteers. Blood reflow was significantly lower in scleroderma patients than in healthy volunteers (p<0.05). This result indicates that the information necessary for assessing blood circulation disorder of peripheral blood vessels in scleroderma patients is objectively obtained by the proposed method.

  14. Whole blood tissue factor procoagulant activity remains detectable during severe aplasia following bone marrow and peripheral blood stem cell transplantation.

    PubMed

    Ozcan, M; Morton, C T; Solovey, A; Dandelet, L; Bach, R R; Hebbel, R P; Slungaard, A; Key, N S

    2001-02-01

    Using a novel whole blood assay, we recently demonstrated that tissue factor procoagulant activity (TF PCA) is present in normal individuals. Preliminary experiments suggested that this activity is localized in the mononuclear cell fraction. Postulating that whole blood TF PCA would therefore be undetectable when monocytes and neutrophils are absent from peripheral blood, we assayed TF PCA during the peri-transplant period in 15 consecutive patients undergoing allogeneic (n = 12) or autologous (n = 3) bone marrow transplantation (BMT) or peripheral blood stem cell transplantation (PBSCT). Baseline (pre-transplant) mean TF PCA was higher in patients compared to normal controls (P <0.005). Unexpectedly, although TF PCA during the period of profound aplasia was significantly reduced compared to baseline (p <0.05), fully 55% of the initial activity remained detectable. During the engraftment phase, TF PCA returned to pre-transplant levels, with a linear correlation between monocyte counts and TF PCA (r = 0.63). In contrast to normal whole blood, incubation of aplastic samples with E. Coli lipopolysaccharide ex vivo failed to induce TF PCA. Throughout the period of study--but especially during the aplastic phase--the absolute number of circulating endothelial cells (CECs) that were TF antigen-positive was increased compared to normals (P <0.001). However, removal of these cells from whole blood samples failed to significantly diminish total TF PCA indicating that CECs alone could not account for the detectable TF PCA during aplasia. We conclude that neither circulating mature myelo-monocytic cells nor endothelial cells can account for all the functionally intact TF in peripheral blood. Further studies are needed to identify the other source(s) of TF PCA.

  15. Peripheral blood gene expression profiles linked to monoamine metabolite levels in cerebrospinal fluid

    PubMed Central

    Luykx, J J; Olde Loohuis, L M; Neeleman, M; Strengman, E; Bakker, S C; Lentjes, E; Borgdorff, P; van Dongen, E P A; Bruins, P; Kahn, R S; Horvath, S; de Jong, S; Ophoff, R A

    2016-01-01

    The blood–brain barrier separates circulating blood from the central nervous system (CNS). The scope of this barrier is not fully understood which limits our ability to relate biological measurements from peripheral to central phenotypes. For example, it is unknown to what extent gene expression levels in peripheral blood are reflective of CNS metabolism. In this study, we examine links between central monoamine metabolite levels and whole-blood gene expression to better understand the connection between peripheral systems and the CNS. To that end, we correlated the prime monoamine metabolites in cerebrospinal fluid (CSF) with whole-genome gene expression microarray data from blood (N=240 human subjects). We additionally applied gene-enrichment analysis and weighted gene co-expression network analyses (WGCNA) to identify modules of co-expressed genes in blood that may be involved with monoamine metabolite levels in CSF. Transcript levels of two genes were significantly associated with CSF serotonin metabolite levels after Bonferroni correction for multiple testing: THAP7 (P=2.8 × 10−8, β=0.08) and DDX6 (P=2.9 × 10−7, β=0.07). Differentially expressed genes were significantly enriched for genes expressed in the brain tissue (P=6.0 × 10−52). WGCNA revealed significant correlations between serotonin metabolism and hub genes with known functions in serotonin metabolism, for example, HTR2A and COMT. We conclude that gene expression levels in whole blood are associated with monoamine metabolite levels in the human CSF. Our results, including the strong enrichment of brain-expressed genes, illustrate that gene expression profiles in peripheral blood can be relevant for quantitative metabolic phenotypes in the CNS. PMID:27959337

  16. Adult peripheral blood mononuclear cells transdifferentiate in vitro and integrate into the retina in vivo.

    PubMed

    Liu, Qian; Guan, Liping; Huang, Bing; Li, Weihua; Su, Qiao; Yu, Minbin; Xu, Xiaoping; Luo, Ting; Lin, Shaochun; Sun, Xuerong; Chen, Mengfei; Chen, Xigu

    2011-06-01

    Adult peripheral blood-derived cells are able to differentiate into a variety of cell types, including nerve cells, liver-like cells and epithelial cells. However, their differentiation into retina-like cells is controversial. In the present study, transdifferentiation potential of human adult peripheral blood mononuclear cells into retina-like cells and integration into the retina of mice were investigated. Freshly isolated adult peripheral blood mononuclear cells were divided into two groups: cells in group I were cultured in neural stem cell medium, and cells in group II were exposed to conditioned medium from rat retinal tissue culture. After 5 days, several distinct cell morphologies were observed, including standard mononuclear, neurons with one or two axons and elongated glial-like cells. Immunohistochemical analysis of neural stem cell, neuron and retina cell markers demonstrated that cells in both groups were nestin-, MAP2 (microtubule-associated protein)- and GFAP (glial fibrillary acidic protein)-positive. Flow cytometry results suggested a significant increase in nestin-, MAP2- and CD16-positive cells in group I and nestin-, GFAP-, MAP2-, vimentin- and rhodopsin-positive cells in group II. To determine survival, migration and integration in vivo, cell suspensions (containing group I or group II cells) were injected into the vitreous or the peritoneum. Tissue specimens were obtained and immunostained 4 weeks after transplantation. We found that cells delivered by intravitreal injection integrated into the retina. Labelled cells were not detected in the retina of mice receiving differentiated cells by intraperitoneal injection, but cells (groups I and II) were detected in the liver and spleen. Our findings revealed that human adult peripheral blood mononuclear cells could be induced to transdifferentiate into neural precursor cells and retinal progenitor cells in vitro, and the differentiated peripheral blood mononuclear cells can migrate and integrate

  17. Effect of transportation on lower respiratory tract contamination and peripheral blood neutrophil function.

    PubMed

    Raidal, S L; Bailey, G D; Love, D N

    1997-06-01

    To evaluate the effect of transportation on lower respiratory tract contamination and peripheral blood neutrophil function in horses and to compare results from transported horses with those obtained in earlier experiments from horses confined with heads elevated. A prospective study. Six horses were transported by road for 12 h. Clinical and haematological examination, transtracheal aspiration and cell function studies were conducted before and after transportation. Results obtained after transportation were compared to pre-transportation values. After transportation, peripheral blood leucocyte and neutrophil numbers were increased and rectal temperatures were evaluated. Transtracheal aspirates showed an accumulation of purulent respiratory tract secretions with increased numbers of bacteria, particularly beta-haemolytic Streptococcus spp and members of the Pasteurellaceae family. Three horses also had increased numbers of bacteria from the Enterobacteriaceae family relative to corresponding samples from earlier studies. Phagocytosis by peripheral blood neutrophils was significantly reduced, while the oxidative burst activity of peripheral blood leucocytes was either unchanged or enhanced. Bacterial contamination of the lower respiratory tract occurs as a routine consequence of transportation of horses and is likely to be an important determinant in the development of transport-associated respiratory disease. Inflammatory airway secretions and increased numbers of bacteria were rapidly cleared, without clinical evidence of significant pulmonary disease and without additional treatment, in normal horses that were allowed to lower their heads after transportation. Peripheral blood neutrophilia and a reduction in neutrophil phagocytic function were evident for at least 36 h after transportation, suggesting that horses may require a number of days to recover from the stress of transportation. As the potential role of bacteria from the Enterobacteriaceae family in the

  18. Peripheral blood Th9 cells and eosinophil apoptosis in asthma patients.

    PubMed

    Hoppenot, Deimantė; Malakauskas, Kęstutis; Lavinskienė, Simona; Bajoriūnienė, Ieva; Kalinauskaitė, Virginija; Sakalauskas, Raimundas

    2015-01-01

    Th9 cells producing interleukin (IL) 9 are novel subset of CD4+ T helper cells, which might contribute to airway inflammation in asthma. Moreover, the effect of IL-9 on eosinophils is still not fully understood. Study aim was to evaluate peripheral blood Th9 cells and eosinophil apoptosis in allergic asthma patients. Eighteen patients with allergic asthma and fourteen patients with allergic rhinitis were examined. Control group included sixteen healthy subjects. Allergic asthma and rhinitis patients did not use corticosteroids and antihistamines at least for 1 week. Peripheral blood eosinophils and CD4(+) cells were isolated by high density gradient centrifugation and magnetic separation. Th9 cells and apoptotic eosinophils were estimated by flow cytometer. Serum IL-9 and IL-5 concentration were determined by ELISA. Peripheral blood Th9 cells percentage was increased in allergic asthma group compared with allergic rhinitis and control group (0.74%±0.32% vs. 0.19%±0.10% and 0.15%±0.08%, respectively, P<0.05). The same tendency was observed for IL-9 (P<0.01). Percentage of peripheral blood apoptotic eosinophils was decreased in allergic asthma and allergic rhinitis groups compared with control group (P<0.05). IL-9 concentration correlated with percentage of Th9 cells (r=0.64, P<0.05) and negatively with percentage of apoptotic eosinophils in allergic asthma group (r=-0.58, P<0.05). Negative correlation was found between apoptotic eosinophils count and IL-5 concentration in allergic asthma group (r=-0.76, P<0.05). Patients with allergic asthma demonstrate increased peripheral blood Th9 cells count and serum IL-9, while eosinophil apoptosis is inversely related to IL-9 concentration. Copyright © 2015 Lithuanian University of Health Sciences. Production and hosting by Elsevier Urban & Partner Sp. z o.o. All rights reserved.

  19. Comparison of gene expression profiles of T cells in porcine colostrum and peripheral blood.

    PubMed

    Ogawa, Shohei; Okutani, Mie; Tsukahara, Takamitsu; Nakanishi, Nobuo; Kato, Yoshihiro; Fukuta, Kikuto; Romero-Pérez, Gustavo A; Ushida, Kazunari; Inoue, Ryo

    2016-09-01

    OBJECTIVE To compare gene expression patterns of T cells in porcine colostrum and peripheral blood. ANIMALS 10 multiparous sows. PROCEDURES Cytotoxic and CD4-CD8 double-positive T cells were separated from porcine colostrum and peripheral blood. Total RNA was extracted. The cDNA prepared from RNA was amplified, labeled, fragmented, and competitively hybridized to DNA microarray slides. The DNA microarray data were validated by use of a real-time reverse-transcription PCR assay, and expression of the genes FOS, NFKBI, IFNG, CXCR6, CCR5, ITGB2, CCR7, and SELL was assessed. Finally, DNA microarray data were validated at the protein level by use of flow cytometry via expression of c-Fos and integrin β-2. RESULTS Evaluation of gene expression profiles indicated that in contrast to results for peripheral blood, numerous cell-signaling pathways might be activated in colostrum. Profile analysis also revealed that FOS and NFKBI (genes of transcription factors) were involved in most cell-signaling pathways and that expression of these genes was significantly higher in colostral T cells than in peripheral blood T cells. Furthermore, CCR7 and SELL (genes of T-cell differentiation markers) in colostral T cells had expression patterns extremely similar to those found in effector or effector memory T cells. CONCLUSIONS AND CLINICAL RELEVANCE All or most of the T cells in colostrum had an effector-like phenotype and thus were more activated than those in peripheral blood. This gene expression profile would enable T cells to migrate to mammary glands, be secreted in colostrum, and likely contribute to passive immunity provided by sows to newborn pigs.

  20. Nuclear receptors expression chart in peripheral blood mononuclear cells identifies patients with Metabolic Syndrome.

    PubMed

    D'Amore, Simona; Vacca, Michele; Graziano, Giusi; D'Orazio, Andria; Cariello, Marica; Martelli, Nicola; Di Tullio, Giuseppe; Salvia, Roberto; Grandaliano, Giuseppe; Belfiore, Anna; Pellegrini, Fabio; Palasciano, Giuseppe; Moschetta, Antonio

    2013-12-01

    Nuclear receptors are a class of 48 ligand-activated transcription factors identified as key players of metabolic and developmental processes. Most of these receptors are potential targets for pharmacological strategies in the Metabolic Syndrome. In the present study, we analyzed changes in the mRNA expression of nuclear receptors in the peripheral blood mononuclear cells of patients with Metabolic Syndrome, in order to identify novel biomarkers of disease and candidate targets for putative therapeutical approaches. We enrolled thirty healthy controls (14 M:16 F) and thirty naïve patients (16 M: 14 F; >3 criteria for Metabolic Syndrome upon Adult Treatment Panel III) without organ damage. Using quantitative real-time PCR, we assessed the expression patterns of nuclear receptors in peripheral blood mononuclear cells. 33/48 nuclear receptors were expressed in peripheral blood mononuclear cells. In patients with Metabolic Syndrome, we found a significant down-regulation of the entire PPAR, NR4A and RAR families, together with a repression of RXRα, VDR, and Rev-Erbα. Furthermore, we performed a novel statistical analysis with classification trees, which allowed us to depict a predictive core of nuclear receptor expression patterns characterizing subjects with Metabolic Syndrome. Random Forest Analysis identified NOR1 and PPARδ, which were both reduced in peripheral blood mononuclear cells and specifically in CD14(+) cells (mostly monocytes), as classifiers of Metabolic Syndrome, with high specificity and sensitivity. Our results point to the use of PPAR and NR4A mRNA levels in the overall peripheral blood mononuclear cells as biomarkers of Metabolic Syndrome and bona fide putative targets of pharmacological therapy. © 2013.

  1. Prognostic significance of immune cells in the tumor microenvironment and peripheral blood of gallbladder carcinoma patients.

    PubMed

    Zhang, Y; Ma, C; Wang, M; Hou, H; Cui, L; Jiang, C; Sun, J; Qu, X

    2017-04-01

    The role of the interaction between tumor cells and inflammatory cells in gallbladder carcinoma (GBC) is unclear. Inflammatory cells exist in both the tumor immune microenvironment and the host peripheral blood circulatory system. In the current study, we examined the prognostic value of inflammatory cells in the tumor microenvironment and peripheral blood in patients with GBC. 98 patients with GBC were recruited in this retrospective study. Using immunohistochemistry, we examined tumor-infiltrating CD3+ generic T-cells, CD8+ cytotoxic T-cells, CD45RO+ memory T-cells, and CD15+ neutrophils. Peripheral venous blood samples were also collected, and absolute neutrophil count (ANC), absolute lymphocyte count (ALC) and neutrophil/lymphocyte ratio (NLR) were measured. The relationships between these variables and patient outcome were evaluated. Survival analysis revealed that the density of CD3+ cell infiltrates in the tumor microenvironment was positively correlated with overall survival (OS) and the density of CD15+ cell infiltrates was negatively correlated with the OS. The combined analysis showed that a high density of CD3+ cell infiltrates combined with a low density of CD15+ cell infiltrates was an independent prognostic factor for GBC. In peripheral blood, survival analysis suggested that ANC and NLR were negatively correlated, while ALC was positively correlated with OS. Multivariate survival analysis showed that NLR was an independent prognostic factor for gallbladder cancer prognosis. The results indicate that the combination of high density of CD3+ cell infiltrates combined with a low density of CD15+ cell infiltrates in tumor samples and pretreatment peripheral blood NLR were independent prognostic factors in patients with GBC.

  2. Genotoxicity of waterpipe smoke in buccal cells and peripheral blood leukocytes as determined by comet assay.

    PubMed

    Al-Amrah, Hadba Jar-Allah; Aboznada, Osama Abdullah; Alam, Mohammad Zubair; ElAssouli, M-Zaki Mustafa; Mujallid, Mohammad Ibrahim; ElAssouli, Sufian Mohamad

    2014-12-01

    Waterpipe smoke causes DNA damage in peripheral blood leukocytes and in buccal cells of smokers. To determine the exposure effect of waterpipe smoke on buccal cells and peripheral blood leukocytes in regard to DNA damage using comet assay. The waterpipe smoke condensates were analyzed by gas chromatography-mass spectrometry (GC-MS). The study was performed on 20 waterpipe smokers. To perform comet assay on bucaal cells of smokers, 10 µl of cell suspension was mixed with 85 µl of pre-warmed 1% low melting agarose, applied to comet slide and electrophoresed. To analyze the effect of smoke condensate in vitro, 1 ml of peripheral blood was mixed with 10 µl of smoke condensate and subjected for comet assay. The GC-MS analysis revealed the presence of 2,3-dihydro-3,5-dihydroxy-6-methyl-4H-pyran-4on, nicotine, hydroxymethyl furancarboxaldehyde and 3-ethoxy-4-hydroxybenzaldehyde in the smoke condensates. Waterpipe smoking caused DNA damage in vivo in buccal cells of smokers. The tail moment and tail length in buccal cells of smokers were 186 ± 26 and 456 ± 71, respectively, which are higher than control. The jurak and moassel smoke condensates were found to cause DNA damage in peripheral blood leukocytes. The moassel smoke condensate was more damaging. There is wide misconception that waterpipe smoking is not as harmful as cigarette smoking. This study demonstrated that waterpipe smoke induced DNA damage in exposed cells. Waterpipe smokes cause DNA damage in buccal cells. The smoke condensate of both jurak and moassel caused comet formation suggesting DNA damage in peripheral blood leukocytes.

  3. Methylene blue modulates transendothelial migration of peripheral blood cells.

    PubMed

    Werner, Isabella; Guo, Fengwei; Bogert, Nicolai V; Stock, Ulrich A; Meybohm, Patrick; Moritz, Anton; Beiras-Fernandez, Andres

    2013-01-01

    Vasoplegia is a severe complication after cardiac surgery. Within the last years the administration of nitric oxide synthase inhibitor methylene blue (MB) became a new therapeutic strategy. Our aim was to investigate the role of MB on transendothelial migration of circulating blood cells, the potential role of cyclic cGMP, eNOS and iNOS in this process, and the influence of MB on endothelial cell apoptosis. Human vascular endothelial cells (HuMEC-1) were treated for 30 minutes or 2 hours with different concentrations of MB. Inflammation was mimicked by LPS stimulation prior and after MB. Transmigration of PBMCs and T-Lymphocytes through the treated endothelial cells was investigated. The influence of MB upon the different subsets of PBMCs (Granulocytes, T- and B-Lymphocytes, and Monocytes) was assessed after transmigration by means of flow-cytometry. The effect of MB on cell apoptosis was evaluated using Annexin-V and Propidium Iodide stainings. Analyses of the expression of cyclic cGMP, eNOS and iNOS were performed by means of RT-PCR and Western Blot. Results were analyzed using unpaired Students T-test. Analysis of endothelial cell apoptosis by MB indicated a dose-dependent increase of apoptotic cells. We observed time- and dose-dependent effects of MB on transendothelial migration of PBMCs. The prophylactic administration of MB led to an increase of transendothelial migration of PBMCs but not Jurkat cells. Furthermore, HuMEC-1 secretion of cGMP correlated with iNOS expression after MB administration but not with eNOS expression. Expression of these molecules was reduced after MB administration at protein level. This study clearly reveals that endothelial response to MB is dose- and especially time-dependent. MB shows different effects on circulating blood cell-subtypes, and modifies the release patterns of eNOS, iNOS, and cGMP. The transendothelial migration is modulated after treatment with MB. Furthermore, MB provokes apoptosis of endothelial cells in a dose

  4. Methylene Blue Modulates Transendothelial Migration of Peripheral Blood Cells

    PubMed Central

    Werner, Isabella; Guo, Fengwei; Bogert, Nicolai V.; Stock, Ulrich A.; Meybohm, Patrick; Moritz, Anton; Beiras-Fernandez, Andres

    2013-01-01

    Vasoplegia is a severe complication after cardiac surgery. Within the last years the administration of nitric oxide synthase inhibitor methylene blue (MB) became a new therapeutic strategy. Our aim was to investigate the role of MB on transendothelial migration of circulating blood cells, the potential role of cyclic cGMP, eNOS and iNOS in this process, and the influence of MB on endothelial cell apoptosis. Human vascular endothelial cells (HuMEC-1) were treated for 30 minutes or 2 hours with different concentrations of MB. Inflammation was mimicked by LPS stimulation prior and after MB. Transmigration of PBMCs and T-Lymphocytes through the treated endothelial cells was investigated. The influence of MB upon the different subsets of PBMCs (Granulocytes, T- and B-Lymphocytes, and Monocytes) was assessed after transmigration by means of flow-cytometry. The effect of MB on cell apoptosis was evaluated using Annexin-V and Propidium Iodide stainings. Analyses of the expression of cyclic cGMP, eNOS and iNOS were performed by means of RT-PCR and Western Blot. Results were analyzed using unpaired Students T-test. Analysis of endothelial cell apoptosis by MB indicated a dose-dependent increase of apoptotic cells. We observed time- and dose-dependent effects of MB on transendothelial migration of PBMCs. The prophylactic administration of MB led to an increase of transendothelial migration of PBMCs but not Jurkat cells. Furthermore, HuMEC-1 secretion of cGMP correlated with iNOS expression after MB administration but not with eNOS expression. Expression of these molecules was reduced after MB administration at protein level. This study clearly reveals that endothelial response to MB is dose- and especially time-dependent. MB shows different effects on circulating blood cell-subtypes, and modifies the release patterns of eNOS, iNOS, and cGMP. The transendothelial migration is modulated after treatment with MB. Furthermore, MB provokes apoptosis of endothelial cells in a dose

  5. Circadian changes of T lymphocyte subsets in human peripheral blood.

    PubMed Central

    Miyawaki, T; Taga, K; Nagaoki, T; Seki, H; Suzuki, Y; Taniguchi, N

    1984-01-01

    The circadian variations in circulating T cell subsets defined by monoclonal antibodies in eight healthy male volunteers were evaluated in whole blood using a flow cytometry. In all subjects, the number of lymphocytes showed a clear rhythmicity with high values at night and low values during the day. This circadian variation in circulating lymphocytes appeared to reflect largely a change in the number of T cells rather than B cells. The percentage of OKT3+ and OKT11+ cells showed a similar fluctuation with a peak at night and a depression during the day. It was found that the percentage of OKT4+ cells varied in parallel with that of T cells, particularly of OKT3+ cells, but the OKT8+ subset was not appreciably altered over a 24 h period. Thus, a circadian variation of T cells could be largely accounted for by a circadian change of OKT4+ cells. Plasma cortisol levels showed an expected circadian variation. It was also shown that there might be an intimate relationship between these circadian changes of T cell subsets and plasma cortisol levels. PMID:6608426

  6. Post-prandial rise of microvesicles in peripheral blood of healthy human donors.

    PubMed

    Suštar, Vid; Bedina-Zavec, Apolonija; Stukelj, Roman; Frank, Mojca; Ogorevc, Eva; Janša, Rado; Mam, Keriya; Veranič, Peter; Kralj-Iglič, Veronika

    2011-03-21

    Microvesicles isolated from body fluids are membrane - enclosed fragments of cell interior which carry information on the status of the organism. It is yet unclear how metabolism affects the number and composition of microvesicles in isolates from the peripheral blood. To study the post - prandial effect on microvesicles in isolates from the peripheral blood of 21 healthy donors, in relation to blood cholesterol and blood glucose concentrations. The average number of microvesicles in the isolates increased 5 hours post - prandially by 52%; the increase was statistically significant (p = 0.01) with the power P = 0.68, while the average total blood cholesterol concentration, average low density lipoprotein cholesterol concentration (LDL-C) and average high density lipoprotein cholesterol concentration (HDL-C) all remained within 2% of their fasting values. We found an 11% increase in triglycerides (p = 0.12) and a 6% decrease in blood glucose (p < 0.01, P = 0.74). The post - prandial number of microvesicles negatively correlated with the post - fasting total cholesterol concentration (r = - 0.46, p = 0.035) while the difference in the number of microvesicles in the isolates between post - prandial and post - fasting states negatively correlated with the respective difference in blood glucose concentration (r = - 0.39, p = 0.05). In a population of healthy human subjects the number of microvesicles in isolates from peripheral blood increased in the post - prandial state. The increase in the number of microvesicles was affected by the fasting concentration of cholesterol and correlated with the decrease in blood glucose.

  7. Peripheral blood stem cell collection in allogeneic donors: impact of venous access.

    PubMed

    Hölig, Kristina; Blechschmidt, Matthias; Kramer, Michael; Zimmer, Kristin; Kroschinsky, Frank; Poppe-Thiede, Kirsten; Bornhäuser, Martin; Ehninger, Gerhard

    2012-12-01

    Peripheral blood stem cell (PBSC) collection is accepted as a routine procedure in related and unrelated healthy donors worldwide. Venous access can be accomplished by peripheral veins or a central venous catheter (CVC). STUDY DESING AND METHODS: We compared efficacy and tolerability of 40 PBSC collections via CVC with 6267 PBSC collections via peripheral veins in healthy allogeneic donors. Results of the leukapheresis procedures and side effects in the donors were evaluated. The median CD34+ cell counts on Day 5 and the results of the stem cell collection were not significantly different between the two groups of allogeneic donors. Pain or problems at the site of puncture or catheter insertion occurred in 58.6% of the donors with a CVC versus 37.8% of the donors with peripheral venous access (p = 0.03). The incidence and severity of paresthesia during the leukapheresis was not significantly different in both groups of donors (p = 0.09). During follow-up no major adverse events related to CVC were reported. Central femoral lines proved to be safe and tolerable in healthy allogeneic donors but peripheral venous access should be preferred, whenever possible. © 2012 American Association of Blood Banks.

  8. Identification of internal control genes for quantitative expression analysis by real-time PCR in bovine peripheral lymphocytes.

    PubMed

    Spalenza, Veronica; Girolami, Flavia; Bevilacqua, Claudia; Riondato, Fulvio; Rasero, Roberto; Nebbia, Carlo; Sacchi, Paola; Martin, Patrice

    2011-09-01

    Gene expression studies in blood cells, particularly lymphocytes, are useful for monitoring potential exposure to toxicants or environmental pollutants in humans and livestock species. Quantitative PCR is the method of choice for obtaining accurate quantification of mRNA transcripts although variations in the amount of starting material, enzymatic efficiency, and the presence of inhibitors can lead to evaluation errors. As a result, normalization of data is of crucial importance. The most common approach is the use of endogenous reference genes as an internal control, whose expression should ideally not vary among individuals and under different experimental conditions. The accurate selection of reference genes is therefore an important step in interpreting quantitative PCR studies. Since no systematic investigation in bovine lymphocytes has been performed, the aim of the present study was to assess the expression stability of seven candidate reference genes in circulating lymphocytes collected from 15 dairy cows. Following the characterization by flow cytometric analysis of the cell populations obtained from blood through a density gradient procedure, three popular softwares were used to evaluate the gene expression data. The results showed that two genes are sufficient for normalization of quantitative PCR studies in cattle lymphocytes and that YWAHZ, S24 and PPIA are the most stable genes. Copyright © 2010 Elsevier Ltd. All rights reserved.

  9. Ultrastructural and Cytochemical Properties of Peripheral Blood Cells of Piebald Naked Carp (Gymnocypris eckloni).

    PubMed

    Zheng, Z X; Tang, Y; Fang, J; Peng, X; Fan, J D; Cui, H M; Yang, L Z

    2017-02-01

    The ultrastructural and cytochemical properties of peripheral blood cells of Gymnocypris eckloni were investigated by transmission electron microscopy and a range of cytochemical techniques to provide clear insight into the structure and function of blood cells from this fish. Ultrastructurally, erythrocytes, leucocytes (neutrophils, eosinophils, lymphocytes, monocytes), thrombocytes and plasma cells were identified in the peripheral blood of G. eckloni. The most special ultrastructural characteristics of blood cells in this fish were that neutrophils exhibited only one type of cytoplasmic granules containing an eccentric, spherical or oval electron-dense core, and eosinophils presented two types of granules with non-uniform electronic density and without crystalloids in their cytoplasm. Neutrophils, eosinophils, lymphocytes, monocytes and thrombocytes were positive for periodic acid-Schiff and α-naphthyl acetate esterase staining. Intense peroxidase positive staining was observed in neutrophils and monocytes, but not in eosinophils, lymphocytes and thrombocytes. Neutrophils, eosinophils and monocytes were stained positively for acid phosphatase, whereas lymphocytes and thrombocytes did not stain. Leucocytes and thrombocytes were negative for alkaline phosphatase and Sudan black B staining. Erythrocytes were negative for all cytochemical staining. The cytochemical and ultrastructural features of peripheral blood cells of G. eckloni were similar to those of other fish species. However, some important differences were identified in G. eckloni. © 2016 Blackwell Verlag GmbH.

  10. Measurement of peripheral B cell subpopulations in common variable immunodeficiency (CVID) using a whole blood method.

    PubMed

    Ferry, B L; Jones, J; Bateman, E A; Woodham, N; Warnatz, K; Schlesier, M; Misbah, S A; Peter, H H; Chapel, H M

    2005-06-01

    Recent reports have described reduced populations of CD27+ memory B cells and increased percentages of undifferentiated B cells in peripheral blood of patients with common variable immunodeficiency (CVID). This work has prompted two attempts to classify CVID based on rapid flow cytometric quantification of peripheral blood memory B cells and immature B cells. Evidence to support the hypothesis that such in vitro B cell classification systems correlate with clinical subtypes of CVID is being sought. For the classification to be useful in routine diagnosis, it is important that the flow cytometric method can be used without prior separation of peripheral blood mononuclear cells (PBMC). We have examined 23 CVID patients and 24 controls, using both PBMC and whole blood, and find an excellent correlation between these methods. The reproducibility of the method was excellent. We classified the CVID patients by all three of the existing classifications, including secretion of immunoglobulin by B cells in vitro as described by Bryant, as well as the more recent flow cytometric classification methods. Only one patient changed classification as a result of using whole blood.

  11. Measurement of peripheral B cell subpopulations in common variable immunodeficiency (CVID) using a whole blood method

    PubMed Central

    Ferry, B L; Jones, J; Bateman, E A; Woodham, N; Warnatz, K; Schlesier, M; Misbah, S A; Peter, H H; Chapel, H M

    2005-01-01

    Recent reports have described reduced populations of CD27+ memory B cells and increased percentages of undifferentiated B cells in peripheral blood of patients with common variable immunodeficiency (CVID). This work has prompted two attempts to classify CVID based on rapid flow cytometric quantification of peripheral blood memory B cells and immature B cells. Evidence to support the hypothesis that such in vitro B cell classification systems correlate with clinical subtypes of CVID is being sought. For the classification to be useful in routine diagnosis, it is important that the flow cytometric method can be used without prior separation of peripheral blood mononuclear cells (PBMC). We have examined 23 CVID patients and 24 controls, using both PBMC and whole blood, and find an excellent correlation between these methods. The reproducibility of the method was excellent. We classified the CVID patients by all three of the existing classifications, including secretion of immunoglobulin by B cells in vitro as described by Bryant, as well as the more recent flow cytometric classification methods. Only one patient changed classification as a result of using whole blood. PMID:15932516

  12. [Making peripheral blood vessels adequate for access in emergency ward services].

    PubMed

    Fraile Fraile, Eloísa; Sánchez Rico, Mercedes; Abejón Arroyo, Dolores; Martínez Fuertes, Carmen

    2008-02-01

    Accessing peripheral blood vessels is one of the most frequently practiced invasive techniques carried out in emergency wards for diagnostic and/or therapeutic purposes. The authors of this article study the peripheral blood vessels and evaluate making their access adequate for treatment by means of a transversal descriptive study. The results of the authors' study show that 33% of the blood vessels catheterized do not comply with the criteria for their adequacy described in the methodology to consider them adequate for catheterization. This relatively high percentage, besides increasing the economic costs in terms of human and material resources, has a negative effect on the functioning of an emergency ward, leading to increased waiting times for other patients pending evaluation-treatment, with the risk that this implies. The elaboration and implementation of a guide to prescribe peripheral blood vessels catheterization may prove useful in order to reduce the number of inadequate blood vessels catheterized and to improve the quality of the care provide by emergency ward personnel.

  13. Inhibition of the activation of Hageman factor (factor XII) by peripheral blood cells.

    PubMed Central

    Ratnoff, O D; Emanuelson, M M; Ziats, N P

    1987-01-01

    Suspensions of peripheral blood mononuclear cells (PBMC), monocytes, T or B lymphocytes, platelets or granulocytes, and cell-depleted supernatant fluids of these suspensions inhibited activation of Hageman factor (HF, Factor XII) by ellagic acid, a property not shared by erythrocytes. PBMC also inhibited HF activation by glass or sulfatides. Contaminating platelets may have contributed to inhibition by PBMC. Elaboration of agents inhibiting HF activation required metabolically active cells. The inhibitor(s) in PBMC supernates were not identified with known agents, but had properties of a nonenzymatic protein. PBMC supernates did not contain fibrinogen, nor alter the thrombin, prothrombin, or partial thromboplastin times of normal plasma, amidolysis by activated plasma thromboplastin antecedent (Factor XIa) or activated Stuart factor (Factor Xa) or esterolysis by C1 (C1 esterase); they inhibited plasmin minimally. These experiments suggest that peripheral blood cells may impede intravascular coagulation. Whether this property helps maintain the fluidity of blood is unclear. PMID:3498741

  14. Expression analysis of psychological stress-associated genes in peripheral blood leukocytes.

    PubMed

    Morita, Kyoko; Saito, Toshiro; Ohta, Masayuki; Ohmori, Tetsuro; Kawai, Kaori; Teshima-Kondo, Shigetada; Rokutan, Kazuhito

    In this study, we have developed a microarray including 1467 cDNAs that were selected to specifically measure stress response in peripheral blood leukocytes. Venous blood was collected from 10 graduate students 2 h before and 2 or 24 h after an open presentation for their Ph.D. The mRNA levels in leukocytes were compared with those prepared 4 weeks before the presentation. Hierarchical cluster showed that distinct groups of genes uniformly changed their expression values in response to the stress. Bayesian t test identified significantly up-regulated 49 genes and down-regulated 21 genes. Most of them are categorized into cytokines, cytokine receptors, growth- or apoptosis-related molecules, and heat shock proteins, suggesting that stressful life events trigger acute responses in leukocytes. Our results suggest that gene expression profile in peripheral blood leukocytes may be a potentially useful method for the assessment of complex stress responses.

  15. Transplacental clearance and blood flows of bovine gravid uterus at several stages of gestation

    SciTech Connect

    Reynolds, L.P.; Ferrell, C.L.

    1987-11-01

    Rates of uterine and umbilical blood flows and transplacental clearance of deuterium oxide (D/sub 2/O) were determined for cows on 137 +/- 1.0 (SE, n = 9), 180 +/- 0.5 (n = 8), 226 +/- 0.4 (n = 9), and 250 +/- 1.8 (n = 5) days of gestation. From days 137 to 250, rates increased 4.5-fold for uterine blood flow, 21-fold for umbilical blood flow, and 14-fold for clearance of D/sub 2/O. Changes in rates of umbilical blood flow and D/sub 2/O clearance paralleled increased rates of fetal growth and metabolism, which have previously been reported to occur during the last half of gestation. The regressions of D/sub 2/O clearance on uterine and umbilical blood flows were significant (P less than 0.01) and explained 94-99% of the variation in placental clearance of D/sub 2/O. Because the rate of D/sub 2/O clearance was always less than that of uterine and umbilical blood flows, and because a relatively simple statistical model explained most of the variation in clearance, it was suggested that a concurrent or countercurrent arrangement of maternal and fetal placental microvasculatures is not adequate to explain clearance of highly diffusable substances across the bovine placenta. In addition, a placental exchange diagram of the data showed the existence of severe uneven distribution of maternal and fetal placental blood flows and/or significant shunting of maternal and fetal placental flows away from areas of exchange. Taken together, these data indicate that the placenta of the cow, like those of the sheep and goat, represents a relatively inefficient system of transplacental exchange.

  16. Effect of hemolysis on nonesterified fatty acid and beta-hydroxybutyrate concentrations in bovine blood.

    PubMed

    Stokol, Tracy; Nydam, Daryl V

    2006-09-01

    Nonesterified fatty acid (NEFA) and beta-hydroxybutyrate (BHB) assays are used for evaluating dairy herds for negative energy balance and subclinical ketosis, respectively. Hemolysis is a common artifact in samples submitted to diagnostic laboratories. The effect of hemolysis on NEFA and BHB in bovine serum was determined. Hemolysis was introduced into 26 serum samples by adding serial dilutions of a red cell hemolysate, prepared by repeated freeze-thawing of EDTA-anticoagulated bovine blood. NEFA, BHB, and degree of hemolysis (hemolytic index) were measured by an automated chemistry analyzer. Two endpoint assays that differed by inclusion of a sample blank were used for NEFA measurement. A kinetic enzymatic assay with 2 reagent sources was used for BHB measurement. The assessed methods yielded similar NEFA or BHB results in baseline, nonhemolyzed samples (median NEFA: 0.25 mEq/L, median BHB: 3 mg/dL, median hemolytic index: 8 units). NEFA results were adversely affected by hemolysis, with values increasing significantly with higher degrees of hemolysis. Median values increased above a critical medical decision limit (0.40 mEq/L) at a hemolytic index of 506 units (marked hemolysis). This increase was prevented by inclusion of a sample blank. Result interpretation was affected in individual animals when samples were moderately hemolyzed (median hemolytic index: 258 units). In contrast, BHB results were unaffected by hemolysis with either reagent source. Thus, assays for measuring NEFAs should include a sample blank and NEFA results should not be interpreted in moderately to markedly hemolyzed bovine samples, because result accuracy cannot be assured.

  17. Immune recognition of exposed xenoantigens on the surface of PEGylated bovine red blood cells.

    PubMed

    Gundersen, Sharon I; Kennedy, Melanie S; Palmer, Andre F

    2008-10-01

    Due to potential problems that can occur during blood transfusion and increasing blood shortages, our group engineered methoxypolyethylene glycol conjugated bovine red blood cells (mPEG-bRBCs) as a potential universal oxygen therapeutic. This current work investigates the immunological properties of mPEG-bRBCs incubated with human plasma (hP) and correlates these properties to exposed Galalpha(1,3)Gal xenoantigens. After mPEG-bRBCs were incubated with hP, the amount of bound IgG and IgM was assessed via flow cytometry. Flow cytometry also assessed the amount of GS-IB4 bound to exposed Galalpha(1,3)Gal xenoantigens. The results of this study demonstrate that most hP samples strongly promote agglutination of mPEG-bRBCs regardless of the extent of mPEG surface coverage or donor blood type. IgG and IgM from hP bound strongly to mPEG-bRBCs. In general, the Galalpha(1,3)Gal xenoantigen remains exposed at all levels of PEG surface coverage. PEGylation did block some of the xenoantigens as the amount of exposed Galalpha (1,3)Gal decreased with increased mPEG surface coverage. However, this was not sufficient to prevent a strong agglutination reaction. Taken together, the results of this study indicate that the current strategy for PEGylating bRBCs is unsatisfactory for the development of immunologically silent oxygen therapeutics.

  18. Probing of Actinobacillus pleuropneumoniae ApxIIIA toxin-dependent cytotoxicity towards mammalian peripheral blood mononucleated cells

    PubMed Central

    Vanden Bergh, Philippe GAC; Zecchinon, Laurent LM; Fett, Thomas; Desmecht, Daniel

    2008-01-01

    Background Actinobacillus pleuropneumoniae, the causative bacterial agent of porcine pleuropneumonia, produces Apx toxins which belong to RTX toxin family and are recognized as the major virulence factors. So far, their target receptor(s) has not been identified and the disease cytopathogenesis remains poorly understood. Production of an active Apx toxin and characterization of its toxic activity constitute the premises necessary to the description of its interaction with a potential receptor. From this point of view, we produced an active recombinant ApxIIIA toxin in order to characterize its toxicity on peripheral blood mononucleated cells (PBMCs) isolated from several species. Findings Toxin preparation exercises a strong cytotoxic action on porcine PBMCs which is directly related to recombinant ApxIIIA since preincubation with polymyxin B does not modify the cytotoxicity rate while preincubation with a monospecific polyclonal antiserum directed against ApxIIIA does. The cell death process triggered by ApxIIIA is extremely fast, the maximum rate of toxicity being already reached after 20 minutes of incubation. Moreover, ApxIIIA cytotoxicity is species-specific because llama, human, dog, rat and mouse PBMCs are resistant. Interestingly, bovine and caprine PBMCs are slightly sensitive to ApxIIIA toxin too. Finally, ApxIIIA cytotoxicity is cell type-specific as porcine epithelial cells are resistant. Conclusion We have produced an active recombinant ApxIIIA toxin and characterized its specific cytotoxicity on porcine PBMCs which will allow us to get new insights on porcine pleuropneumonia pathogenesis in the future. PMID:19046441

  19. Tissue engineering with peripheral blood-derived mesenchymal stem cells promotes the regeneration of injured peripheral nerves.

    PubMed

    Pan, Mengjie; Wang, Xianghai; Chen, Yijing; Cao, Shangtao; Wen, Jinkun; Wu, Guofeng; Li, Yuanyuan; Li, Lixia; Qian, Changhui; Qin, Zhenqi; Li, Zhenlin; Tan, Dandan; Fan, Zhihao; Wu, Wutian; Guo, Jiasong

    2017-06-01

    Peripheral nerve injury repair can be enhanced by Schwann cell (SC) transplantation, but clinical applications are limited by the lack of a cell source. Thus, alternative systems for generating SCs are desired. Herein, we found the peripheral blood-derived mesenchymal stem cells (PBMSCs) could be induced into SC like cells with expressing SC-specific markers (S100, P75NTR and CNPase) and functional factors (NGF, NT-3, c-Fos, and Krox20). When the induced PBMSCs (iPBMSCs) were transplanted into crushed rat sciatic nerves, they functioned as SCs by wrapping the injured axons and expressing myelin specific marker of MBP. Furthermore, iPBMSCs seeded in an artificial nerve conduit to bridge a 10-mm defect in a sciatic nerve achieved significant nerve regeneration outcomes, including axonal regeneration and remyelination, nerve conduction recovery, and restoration of motor function, and attenuated myoatrophy and neuromuscular junction degeneration in the target muscle. Overall, the data from this study indicated that PBMSCs can transdifferentiate towards SC-like cells and have potential as grafting cells for nerve tissue engineering. Copyright © 2017. Published by Elsevier Inc.

  20. Comparative study of peripheral blood smear, quantitative buffy coat and modified centrifuged blood smear in malaria diagnosis.

    PubMed

    Bhandari, P L; Raghuveer, C V; Rajeev, A; Bhandari, P D

    2008-01-01

    The present study was aimed at modifying the centrifuged blood smear (modified centrifuged blood smear or MCBS), to make it a feasible and standardized procedure. The results obtained were compared with the current diagnostic methods - peripheral blood smear (PBS) and quantitative buffy coat (QBC). Blood samples collected from 100 suspected malaria patients were subjected to all three tests. It was found that PBS had 86.79% sensitivity and was absolutely specific. QBC was 96.22% sensitive and 93.61% specific. The majority of variations occurred in PBS negative cases; cases with parasite count Plasmodium falciparum. It was seen that by the addition of centrifugation to the conventional smear technique (MCBS) improved its sensitivity from 86.79% to near 100%. QBC and MCBS were found superior to PBS. Since MCBS combines principles of both QBC and PBS, it is as sensitive as QBC, as specific as PBS, and above all, easily performed and affordable.

  1. Infusion of hemolyzed red blood cells within peripheral blood stem cell grafts in patients with and without sickle cell disease.

    PubMed

    Fitzhugh, Courtney D; Unno, Hayato; Hathaway, Vincent; Coles, Wynona A; Link, Mary E; Weitzel, R Patrick; Zhao, Xiongce; Wright, Elizabeth C; Stroncek, David F; Kato, Gregory J; Hsieh, Matthew M; Tisdale, John F

    2012-06-14

    Peripheral blood stem cell (PBSC) infusions are associated with complications such as elevated blood pressure and decreased creatinine clearance. Patients with sickle cell disease experience similar manifestations, and some have postulated release of plasma-free hemoglobin with subsequent nitric oxide consumption as causative. We sought to evaluate whether the infusion of PBSC grafts containing lysed red blood cells (RBCs) leads to the toxicity observed in transplant subjects. We report a prospective cohort study of 60 subjects divided into 4 groups based on whether their infusions contained dimethyl sulfoxide (DMSO) and lysed RBCs, no DMSO and fresh RBCs, DMSO and no RBCs, or saline. Our primary end point, change in maximum blood pressure compared with baseline, was not significantly different among groups. Tricuspid regurgitant velocity and creatinine levels also did not differ significantly among groups. Our data do not support free hemoglobin as a significant contributor to toxicity associated with PBSC infusions. This study was registered at clinicaltrials.gov (NCT00631787).

  2. Stability of endogenous GHB in vitreous humor vs peripheral blood in dead bodies.

    PubMed

    Busardò, Francesco Paolo; Mannocchi, Giulio; Giorgetti, Raffaele; Pellegrini, Manuela; Baglio, Giovanni; Zaami, Simona; Marinelli, Enrico; Pichini, Simona

    2017-05-01

    For the first time, the stability of GHB was tested in post-mortem peripheral blood and vitreous humor samples, collected from 22 dead bodies at two different times: at the external body examination at the place of death and then during autopsy. An ad hoc method for the detection and quantification of GHB in vitreous humor by gas chromatography coupled to mass spectrometry (GC-MS) was developed and validated, with a good linearity between 0.1 and 50μg/mL (r(2)=0.991) and a precision and accuracy always better than 10% and an analytical recovery higher than 90%. The geometric mean of GHB concentration in the 22 peripheral blood samples at t0 was: 3.6μg/mL (95% CI: 2.3-5.9μg/mL) and at t1 it was 7.4μg/mL (95% CI: 5.0-10.9μg/mL); that of GHB in the 22 vitreous humor at t0 was: 2.5μg/mL (95% CI: 1.5-4.1μg/mL) and at t1 it was 3.0μg/mL (95% CI: 1.9-4.8μg/mL). There was no significant difference between the GHB concentrations in vitreous humor and peripheral blood at t0 in all the samples (p>0.10). Conversely at t1, the increase of GHB in the peripheral blood was significantly increased by a 102% (range: 86-120%) (p<0.001 vs t0), while in the vitreous humor only a slight increase by 19% was observed (range: 16-21%) (p>0.05 vs t0). Finally at t1, GHB values in the two matrices were statistically different, being that of peripheral blood higher (p<0.01). This study demonstrated the usefulness of vitreous humor as a more stable alternative matrix in comparison to peripheral blood for the post-mortem determination of endogenous GHB. Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.

  3. Donor cell-derived acute myeloblastic leukemia after allogeneic peripheral blood hematopoietic stem cell transplantation for juvenile myelomonocytic leukemia.

    PubMed

    Cetin, Zafer; Tezcan, Gulsun; Karauzum, Sibel Berker; Kupesiz, Alphan; Manguoglu, Ayse Esra; Yesilipek, Akif; Luleci, Guven; Hazar, Volkan

    2006-11-01

    Despite its rarity, donor cell leukemia (DCL) is a most intriguing entity. We report here the case of a 5 year-old girl with juvenile myelomonocytic leukemia and normal female karyotype who developed acute myeloblastic leukemia with a karyotype of 46, X, t(X; 7) (p21; p11.2), der(7) t(3; 7) (q13.3; q22) 5 months after peripheral blood hematopoietic stem cell transplantation from her HLA-matched sister. We performed the analysis of short tandem repeat sequence markers to DNA obtained from donor peripheral blood, patient's peripheral blood including leukemic blasts and patient's hair root. This analysis showed that the leukemic blood DNA matched the donor blood DNA and not the patient's DNA, thus confirming DCL. To our knowledge, this is the first case of DCL after peripheral blood SCT for juvenile myelomonocytic leukemia.

  4. Profiling of exercise-induced transcripts in the peripheral blood cells of Thoroughbred horses

    PubMed Central

    TOZAKI, Teruaki; KIKUCHI, Mio; KAKOI, Hironaga; HIROTA, Kei-ichi; MUKAI, Kazutaka; AIDA, Hiroko; NAKAMURA, Seiji; NAGATA, Shun-ichi

    2016-01-01

    ABSTRACT Transcriptome analyses based on DNA microarray technology have been used to investigate gene expression profiles in horses. In this study, we aimed to identify exercise-induced changes in the expression profiles of genes in the peripheral blood of Thoroughbred horses using DNA microarray technology (15,429 genes on 43,603 probes). Blood samples from the jugular vein were collected from six horses before and 1 min, 4 hr, and 24 hr after all-out running on a treadmill. After the normalization of microarray data, a total of 26,830 probes were clustered into four groups and 11 subgroups showing similar expression changes based on k-mean clustering. The expression level of inflammation-related genes, including interleukin-1 receptor type II (IL-1R2), matrix metallopeptidase 8 (MMP8), protein S100-A8 (S100-A8), and serum amyloid A (SAA), increased at 4 hr after exercise, whereas that of c-Fos (FOS) increased at 1 min after exercise. These results indicated that the inflammatory response increased in the peripheral blood cells after exercise. Our study also revealed the presence of genes that may not be affected by all-out exercise. In conclusion, transcriptome analysis of peripheral blood cells could be used to monitor physiological changes induced by various external stress factors, including exercise, in Thoroughbred racehorses. PMID:27974875

  5. Prognostic Value of RT-PCR Tyrosinase Detection in Peripheral Blood of Melanoma Patients

    PubMed Central

    Carrillo, Esmeralda; Prados, José; Marchal, Juan Antonio; Boulaiz, Houria; Martínez, Antonio; Rodríguez-Serrano, Fernando; Caba, Octavio; Serrano, Salvio; Aránega, Antonia

    2006-01-01

    Malignant melanoma (MM) prognosis has been related to tumour thickness and clinical stage and metastasis risk has been associated with presence of tumour cells in peripheral blood. The aim of this study was to determine the relationship between presence of tyrosinase in peripheral blood of MM patients and their clinical prognosis. Blood samples from 58 MM patients (stage I–IV) were analysed, using RT-PCR assay to detect tyrosinase mRNA. The results showed that positive RT-PCR assay for tyrosinase were significantly associated with clinical status and tumour thickness. After a median follow-up of 24 months, RT-PCR results were found to be significant correlated with recurrence (p < 0.05) and clinical stage III (p < 0.05). Separate analysis of stage III tumours to determine the prognostic value of tyrosinase presence in peripheral blood showed an overall 24-month survival rate of 70% in the RT-PCR negative group versus 10% in the positive group (p < 0.02). These results suggest that detection of circulating melanoma cells may be especially relevant in stage III patients, in whom RT-PCR positivity defines a subpopulation at high risk of recurrence. PMID:16788251

  6. Arginase activity in peripheral blood of patients with intestinal schistosomiasis, Wonji, Central Ethiopia.

    PubMed

    Getaneh, A; Tamrat, A; Tadesse, K

    2015-07-01

    Morbidity and mortality caused by schistosomiasis usually results from immunopathology. But the underlying mechanisms are not yet clearly understood. Th2-type immune response is thought to be dominant during chronic schistosomiasis, and upregulation of arginase-I is one component of this milieu. A cohort study was conducted to assess arginase activity in peripheral blood of humans with intestinal schistosomiasis in Wonji-Shoa Sugar Estate, Central Ethiopia. Laboratory-confirmed 30 Schistosoma mansoni-infected patients and 18 apparently healthy controls were recruited. Faecal egg count was carried out by Kato-Katz technique. Plasma and peripheral blood mononuclear cells (PBMCs) were isolated from whole blood. Activity of arginase in plasma and PBMC lysates was measured, and results were compared with that of controls. Twenty-one of 30 patients had light infection, whereas moderate and heavy intensity infections were observed in eight and only one patient(s), respectively. A significant increase in both PBMC (patients: 59.96 + 82.99, controls: 25.44 + 24.6 mU/mg protein, P < 0.0001) and plasma (patients: 1.61 + 2.19, controls: 0.31 + 0.73 mU/mL plasma, P < 0.0001) arginase activity was observed during human S. mansoni infection. Arginase activity increases in peripheral blood of patients with intestinal schistosomiasis.

  7. Matrix metalloproteinase-3 mRNA: a promising peripheral blood marker for diagnosis of endometriosis.

    PubMed

    De Sanctis, Paola; Elmakky, Amira; Farina, Antonio; Caramelli, Elisabetta; Seracchioli, Renato; Mabrouk, Mohamed; Mignemi, Giuseppe; Venturoli, Stefano; Villa, Gioia; Guerrini, Manuela; Manuzzi, Linda; Montanari, Giulia; Valvassori, Luisa; Zucchini, Cinzia

    2011-01-01

    Endometriosis is an invasive disease. Its diagnosis depends on laparoscopy, which is traumatic and associated with potential complications. The aim of this study was to develop a rapid, reliable, and less invasive diagnostic test for endometriosis. We hypothesized that genes related to cell invasion would be transcriptionally upregulated in endometriosis, and tested whether blood levels of their transcripts might be used as biomarkers of endometriosis. We used quantitative real-time reverse transcription polymerase chain reaction (qRT-PCR) to quantify the mRNA levels of vascular endothelial growth factor A (VEGFA), matrix metalloproteinase-3 (MMP-3), and MMP-9 in peripheral blood from 20 patients with mild/intermediate endometriosis, 20 patients with severe endometriosis and 20 endometriosis-free subjects. Our results indicate that circulating mRNA for MMP-3 is significantly higher in patients with endometriosis than in control patients, regardless of the degree of severity. Conversely, the level of circulating mRNA for VEGFA and MMP-9 did not distinguish patients from controls. MMP-3 mRNA is a promising peripheral blood marker that discriminates between patients with endometriosis and healthy subjects. Our results support the possibility of finding genes suitable for diagnostic qRT-PCR for endometriosis in peripheral blood and should be explored further. Copyright © 2010 S. Karger AG, Basel.

  8. Anxiety and cerebral blood flow during behavioral challenge. Dissociation of central from peripheral and subjective measures

    SciTech Connect

    Zohar, J.; Insel, T.R.; Berman, K.F.; Foa, E.B.; Hill, J.L.; Weinberger, D.R.

    1989-06-01

    To investigate the relationship between anxiety and regional cerebral blood flow, we administered behavioral challenges to 10 patients with obsessive-compulsive disorder while measuring regional cerebral blood flow with the xenon 133 inhalation technique. Each patient was studied under three conditions: relaxation, imaginal flooding, and in vivo (actual) exposure to the phobic stimulus. Subjective anxiety, obsessive-compulsive ratings, and autonomic measures (heart rate, blood pressure) increased significantly, but respiratory rate and PCO/sub 2/ did not change across the three conditions. Regional cerebral blood flow increased slightly (in the temporal region) during imaginal flooding, but decreased markedly in several cortical regions during in vivo exposure, when anxiety was highest by subjective and peripheral autonomic measures. These results demonstrate that intense anxiety can be associated with decreased rather than increased cortical perfusion and that ostensibly related states of anxiety (eg, anticipatory and obsessional anxiety) may be associated with opposite effects on regional cerebral blood flow.

  9. Relationship between zinc malnutrition and alterations in murine peripheral blood leukocytes

    SciTech Connect

    King, L.E.; Morford, L.A.; Fraker, P.J. )

    1991-03-15

    Studies using a murine model have shown that the immune system responds rapidly and adversely to zinc deficiency. The extent of alteration of peripheral blood leukocytes (PBL) and immunoglobulin levels were investigated in four zinc dietary groups: zinc adequate (ZA); restricted fed zinc adequate (RZA); marginal zinc deficient (MZD, 72-76% of ZA mouse weight); and severely zinc deficient. The peripheral white blood cell count was 3.66 {plus minus} 1.08 {times} 10{sup 6} cells/ml for ZA mice decreasing by 21%, 28% and 54% for RZA, MZD and SZD mice respectively. An equally dramatic change in the flow cytometric light scatter profile was found. ZA mice had 66% lymphocytes and 21% polymorphonuclear granulocytes (PMN) in their peripheral blood while MZD and SZD mice contained 43% and 30% lymphocytes and 40% and 60% PMNs respectively. Analysis of the phenotypic distribution of specific classes of lymphocytes revealed ZA blood contained 25% B-cells and 40% T-cells (CD5{sup +}). B-cells decreased 40-50% for RZA and MZD mice and 60-70% for SZD mice. The decline in CD5{sup +} T-cells was more modest at 30% and 45% for MZD and SZD mice. A nearly 40% decline in both T{sub h} and T{sub c/s} cells was noted for both MZD and SZD mice. Radioimmunoassay of serum for changes in IgM and IgG content revealed no change among dietary groups while serum zinc decreased 10% for RZA mice and 50% for both MZD and SZD mice. The authors conclude that peripheral blood differential counts in concert with total B and T-cell phenotype may serve as indicators of zinc status while serum zinc and Ig will not.

  10. Smooth muscle progenitor cells from peripheral blood promote the neovascularization of endothelial colony-forming cells

    SciTech Connect

    Joo, Hyung Joon; Seo, Ha-Rim; Jeong, Hyo Eun; Choi, Seung-Cheol; Park, Jae Hyung; Yu, Cheol Woong; Hong, Soon Jun; Chung, Seok; Lim, Do-Sun

    2014-07-11

    Highlights: • Two distinct vascular progenitor cells are induced from adult peripheral blood. • ECFCs induce vascular structures in vitro and in vivo. • SMPCs augment the in vitro and in vivo angiogenic potential of ECFCs. • Both cell types have synergistic therapeutic potential in ischemic hindlimb model. - Abstract: Proangiogenic cell therapy using autologous progenitors is a promising strategy for treating ischemic disease. Considering that neovascularization is a harmonized cellular process that involves both endothelial cells and vascular smooth muscle cells, peripheral blood-originating endothelial colony-forming cells (ECFCs) and smooth muscle progenitor cells (SMPCs), which are similar to mature endothelial cells and vascular smooth muscle cells, could be attractive cellular candidates to achieve therapeutic neovascularization. We successfully induced populations of two different vascular progenitor cells (ECFCs and SMPCs) from adult peripheral blood. Both progenitor cell types expressed endothelial-specific or smooth muscle-specific genes and markers, respectively. In a protein array focused on angiogenic cytokines, SMPCs demonstrated significantly higher expression of bFGF, EGF, TIMP2, ENA78, and TIMP1 compared to ECFCs. Conditioned medium from SMPCs and co-culture with SMPCs revealed that SMPCs promoted cell proliferation, migration, and the in vitro angiogenesis of ECFCs. Finally, co-transplantation of ECFCs and SMPCs induced robust in vivo neovascularization, as well as improved blood perfusion and tissue repair, in a mouse ischemic hindlimb model. Taken together, we have provided the first evidence of a cell therapy strategy for therapeutic neovascularization using two different types of autologous progenitors (ECFCs and SMPCs) derived from adult peripheral blood.

  11. A Semi-automated Approach to Preparing Antibody Cocktails for Immunophenotypic Analysis of Human Peripheral Blood

    PubMed Central

    Koguchi, Yoshinobu; Gonzalez, Iliana L.; Meeuwsen, Tanisha L.; Miller, William L.; Haley, Daniel P.; Tanibata-Branham, Alice N.; Bahjat, Keith S.

    2016-01-01

    Immunophenotyping of peripheral blood by flow cytometry determines changes in the frequency and activation status of peripheral leukocytes during disease and treatment. It has the potential to predict therapeutic efficacy and identify novel therapeutic targets. Whole blood staining utilizes unmanipulated blood, which minimizes artifacts that can occur during sample preparation. However, whole blood staining must also be done on freshly collected blood to ensure the integrity of the sample. Additionally, it is best to prepare antibody cocktails on the same day to avoid potential instability of tandem-dyes and prevent reagent interaction between brilliant violet dyes. Therefore, whole blood staining requires careful standardization to control for intra and inter-experimental variability. Here, we report deployment of an automated liquid handler equipped with a two-dimensional (2D) barcode reader into a standard process of making antibody cocktails for flow cytometry. Antibodies were transferred into 2D barcoded tubes arranged in a 96 well format and their contents compiled in a database. The liquid handler could then locate the source antibody vials by referencing antibody names within the database. Our method eliminated tedious coordination for positioning of source antibody tubes. It provided versatility allowing the user to easily change any number of details in the antibody dispensing process such as specific antibody to use, volume, and destination by modifying the database without rewriting the scripting in the software method for each assay. A proof of concept experiment achieved outstanding inter and intra- assay precision, demonstrated by replicate preparation of an 11-color, 17-antibody flow cytometry assay. These methodologies increased overall throughput for flow cytometry assays and facilitated daily preparation of the complex antibody cocktails required for the detailed phenotypic characterization of freshly collected anticoagulated peripheral blood

  12. The detection of melanoma cells in peripheral blood by reverse transcription-polymerase chain reaction.

    PubMed Central

    Foss, A. J.; Guille, M. J.; Occleston, N. L.; Hykin, P. G.; Hungerford, J. L.; Lightman, S.

    1995-01-01

    Both cutaneous and uveal melanoma undergo haematogenous dissemination. Detection of tyrosinase mRNA by reverse transcription-polymerase chain reaction (RT-PCR) has been described as an extremely sensitive way of detecting circulating viable melanoma cells in the peripheral venous blood, and this technique may be of value in the early detection of dissemination. Also, it has been suggested that surgical manipulation of the eye, such as occurs during enucleation, can provoke uveal melanoma dissemination. The purpose of this study was to evaluate whether tyrosinase mRNA is detectable in the peripheral blood of patients with uveal and cutaneous melanoma and in patients with uveal melanoma undergoing surgical procedures on the eye harbouring the tumour. Venous blood samples from 36 patients diagnosed as having active uveal melanoma and from six patients with advanced metastatic cutaneous melanoma were analysed. In addition, blood samples were spiked with known numbers of cells from three cell lines and four primary uveal melanoma cultures. The reported sensitivity of the technique was confirmed, with an ability to detect down to one cell per ml of blood. All 51 blood samples from the 36 patients with uveal melanoma were negative, and this included 20 perioperative blood samples. The test was also negative for the six patients with advanced cutaneous melanoma. There were two positives among 31 control samples analysed. This study demonstrates that there are far fewer circulating viable melanocytes than has been previously supposed in patients with melanoma and that the RT-PCR is of no clinical value in detecting metastatic melanoma disease. There was no evidence for surgery causing a bolus of melanoma cells to enter the peripheral circulation. Images Figure 1 Figure 2 PMID:7599046

  13. Alteration of immunologic responses on peripheral blood in the acute phase of ischemic stroke: blood genomic profiling study.

    PubMed

    Oh, Seung-Hun; Kim, Ok-Joon; Shin, Dong-Ah; Song, Jihwan; Yoo, Hanna; Kim, Yu-Kyung; Kim, Jin-Kyeoung

    2012-08-15

    Peripheral blood cells and inflammatory mediators have a detrimental effect on brain during cerebral ischemia. We investigated the immunologic changes on peripheral blood in the acute phase of ischemic stroke using RNA microarray. mRNA microarray and real time-polymerase chain reaction (RT-PCR) for genes of interest in microarray data were analyzed in 12 stroke patients and 12 controls. Plasma matrix metalloproteinase-9 (MMP-9) concentrations were measured in 120 stroke patients and 82 controls. In microarray analysis, a total of 11 genes of interest showed different expression in patients with ischemic stroke. The three most highly expressed genes were C19orf59 (chromosome 19 open reading frame 59), MMP9 and IL18RAP (interleukin-18 receptor accessory protein), whereas gene with the lowest expression was GNLY (granulysin). The expression patterns of three selected genes (MMP9, IL18RAP and GNLY) were validated by RT-PCR. The plasma concentration of MMP-9 was significantly elevated in the stroke patients, and showed a weakly positive correlation with infarct volume. Gene set enrichment analysis (GSEA) showed that gene sets related to immunity and defense, signal transduction, transport and cell adhesion were significant in acute ischemic stroke. In the peripheral blood, numerous genes of inflammatory mediators, including MMP9, IL18RAP and GNLY, are altered in the acute phase of ischemic stroke. This stroke-specific gene expression profiling provides valuable information about the role of peripheral inflammation to the pathophysiological mechanism of ischemic stroke. Copyright © 2012 Elsevier B.V. All rights reserved.

  14. [Structure of red blood cell and peripheral blood lymphocytes membranes in children--residents of contaminated areas in the remote period of Chernobyl].

    PubMed

    Stepanova, E I; Vdovenko, V Iu; Litvinets, O M

    2013-06-01

    We applied scanning electron microscope to study of surface architectonics of erythrocytes and lymphocytes peripheral blood in children born after the Chernobyl accident and living in conditions of chronic incorporation 137Cs. We found significant changes in surface structure membranes of red blood cells and peripheral blood lymphocytes in the basic childrens group compared with control one. The most striking changes were in children with levels incorporated 137Cs from 6845 to 16522 Bq.

  15. Inter-Individual Differences in RNA Levels in Human Peripheral Blood

    PubMed Central

    Chomczynski, Piotr; Wilfinger, William W.; Eghbalnia, Hamid R.; Kennedy, Amy; Rymaszewski, Michal; Mackey, Karol

    2016-01-01

    Relatively little is known about the range of RNA levels in human blood. This report provides assessment of peripheral blood RNA level and its inter-individual differences in a group of 35 healthy humans consisting of 25 females and 10 males ranging in age from 50 to 89 years. In this group, the average total RNA level was 14.59 μg/ml of blood, with no statistically significant difference between females and males. The individual RNA level ranged from 6.7 to 22.7 μg/ml of blood. In healthy subjects, the repeated sampling of an individual’s blood showed that RNA level, whether high or low, was stable. The inter-individual differences in RNA level in blood can be attributed to both, differences in cell number and the amount of RNA per cell. The 3.4-fold range of inter-individual differences in total RNA levels, documented herein, should be taken into account when evaluating the results of quantitative RT-PCR and/or RNA sequencing studies of human blood. Based on the presented results, a comprehensive assessment of gene expression in blood should involve determination of both the amount of mRNA per unit of total RNA (U / ng RNA) and the amount of mRNA per unit of blood (U / ml blood) to assure a thorough interpretation of physiological or pathological relevance of study results. PMID:26863434

  16. Calcitonin gene-related peptide in peripheral blood as a biomarker for migraine.

    PubMed

    Ramón, César; Cernuda-Morollón, Eva; Pascual, Julio

    2017-06-01

    There is no available biomarker for any of the primary headaches, including migraine. As demonstrated in jugular blood, during a migraine attack, trigeminal activation releases several neuropeptides, very especially calcitonin gene-related peptide (CGRP), which gives rise to the typical throbbing migraine pain. Here, we review the current evidence for measurement of peripheral CGRP levels as a potential biomarker for trigemino-vascular activation in migraine. Several studies, including a limited number of migraine patients, have shown increased peripheral CGRP levels during migraine attacks. The maximum increase in plasma CGRP levels was reached within 2 h of the onset of the attacks and can be reverted by triptans. In addition, CGRP levels measured in peripheral blood outside migraine attacks and in the absence of symptomatic medication have been shown to be increased in chronic migraine patients. Increased CGRP levels were able to predict the response to onabotulinumtoxinA treatment and were reduced 1 month after onabotulinumtypeA therapy. Although CGRP data must be confirmed and expanded in future studies and specificity of CGRP levels should be studied in entities able to resemble migraine, it seems that peripheral CGRP levels are a good biomarker of acute migraine and somewhat specific and sensitive interictally in chronic migraine.

  17. Peripheral blood T- and B-cell immunophenotypic abnormalities in selected women with unexplained recurrent miscarriage.

    PubMed

    Carbone, Javier; Sarmiento, Elizabeth; Gallego, Antonio; Lanio, Nallibe; Navarro, Joaquin; García, Sandra; Fernandez-Cruz, Eduardo

    2016-02-01

    We aimed to investigate if women with recurrent miscarriage disclosed abnormalities in the maturation and activation status of peripheral blood lymphocyte subsets. In a case control study, 24 women with recurrent miscarriage, 37 women with children but no history of miscarriage and 39 women without previous pregnancies were evaluated. Lymphocyte subsets were evaluated using three-colour flow-cytometry. Selected women with recurrent miscarriage had significantly higher absolute counts of central memory CD4+ T-cells, CD8+DR+ T-cells and memory non-switched B-cells than the control groups. Recurrent miscarriage may be associated with abnormalities of the maturation and activation status of peripheral blood T and B lymphocytes.

  18. Peripheral arterial blood pressure monitoring adequately tracks central arterial blood pressure in critically ill patients: an observational study.

    PubMed

    Mignini, Mariano Alejandro; Piacentini, Enrique; Dubin, Arnaldo

    2006-01-01

    Invasive arterial blood pressure monitoring is a common practice in intensive care units (ICUs). Accuracy of invasive blood pressure monitoring is crucial in evaluating the cardiocirculatory system and adjusting drug therapy for hemodynamic support. However, the best site for catheter insertion is controversial. Lack of definitive information in critically ill patients makes it difficult to establish guidelines for daily practice in intensive care. We hypothesize that peripheral and central mean arterial blood pressures are interchangeable in critically ill patients. This is a prospective, observational study carried out in a surgical-medical ICU in a teaching hospital. Fifty-five critically ill patients with clinical indication of invasive arterial pressure monitoring were included in the study. No interventions were made. Simultaneous measurements were registered in central (femoral) and peripheral (radial) arteries. Bias and precision between both measurements were calculated with Bland-Altman analysis for the whole group. Bias and precision were compared between patients receiving high doses of vasoactive drugs (norepinephrine or epinephrine >0.1 microg/kg/minute or dopamine >10 microg/kg/minute) and those receiving low doses (norepinephrine or epinephrine <0.1 microg/kg/minute or dopamine <10 microg/kg/minute). Central mean arterial pressure was 3 +/- 4 mmHg higher than peripheral mean arterial pressure for the whole population and there were no differences between groups (3 +/- 4 mmHg for both groups). Measurement of mean arterial blood pressure in radial or femoral arteries is clinically interchangeable. It is not mandatory to cannulate the femoral artery, even in critically ill patients receiving high doses of vasoactive drugs.

  19. A strategy to identify genes associated with circulating solid tumor cell survival in peripheral blood.

    PubMed Central

    Fournier, M. V.; Carvalho, M. G.; Pardee, A. B.

    1999-01-01

    Efforts in metastasis research have centered on the phenotypic and genetic differences between primary site and metastatic site tumors. However, genes that may be used as molecular markers of metastasis in circulating tumor cells remain unidentified. Genes regulating the dissemination and survival of solid tumor cells in the blood, as well as their adaptation to new environments, could be candidates for unique metastatic tumor markers. Differential display (DD) was conducted to compare the blood of tumor-free individuals with the blood of patients with lung, breast, and colon cancers. Twenty-one up-expressed genes in the tumor patient blood samples but none in the tumor-free donor blood samples were identified. Nine of these samples were isolated, amplified, and directly sequenced. A gene AB-1 homologous to a Bcl-2 family member, which might function as an apoptosis inhibitor, was identified. The overexpression of an apoptosis inhibitor in blood from patients with metastatic tumors might be correlated with the capability of solid tumor cells to survive in peripheral blood. This is the first demonstration of the usefulness of comparing control and patient blood samples by DD to find novel potential genetic markers identifying metastasis in the blood. http://link.springer-ny. com/link/service/journals/00020/bibs/5n5p313.html Images Fig. 1 Fig. 2 Fig. 3 PMID:10390547

  20. Correlates of Peripheral Blood Mitochondrial DNA Content in a General Population

    PubMed Central

    Knez, Judita; Winckelmans, Ellen; Plusquin, Michelle; Thijs, Lutgarde; Cauwenberghs, Nicholas; Gu, Yumei; Staessen, Jan A.; Nawrot, Tim S.; Kuznetsova, Tatiana

    2016-01-01

    Accumulation of mitochondrial DNA (mtDNA) mutations leads to alterations of mitochondrial biogenesis and function that might produce a decrease in mtDNA content within cells. This implies that mtDNA content might be a potential biomarker associated with oxidative stress and inflammation. However, data on correlates of mtDNA content in a general population are sparse. Our goal in the present study was to describe in a randomly recruited population sample the distribution and determinants of peripheral blood mtDNA content. From 2009 to 2013, we examined 689 persons (50.4% women; mean age = 54.4 years) randomly selected from a Flemish population (Flemish Study on Environment, Genes, and Health Outcomes). Relative mtDNA copy number as compared with nuclear DNA was measured by quantitative real-time polymerase chain reaction in peripheral blood. There was a curvilinear relationship between relative mtDNA copy number and age. mtDNA content slightly increased until the fifth decade of life and declined in older subjects (Page2 = 0.0002). mtDNA content was significantly higher in women (P = 0.007) and increased with platelet count (P < 0.0001), whereas it was inversely associated with white blood cell count (P < 0.0001). We also observed lower mtDNA content in women using estroprogestogens (P = 0.044). This study demonstrated in a general population that peripheral blood mtDNA content is significantly associated with sex and age. Blood mtDNA content is also influenced by platelet and white blood cell counts and estroprogestogen intake. Further studies are required to clarify the impact of chronic inflammation and hormone therapy on mitochondrial function. PMID:26702630

  1. Monitoring of Epstein-Barr Virus DNA Load in Peripheral Blood by Quantitative Competitive PCR

    PubMed Central

    Stevens, Servi J. C.; Vervoort, Marcel B. H. J.; van den Brule, Adriaan J. C.; Meenhorst, Pieter L.; Meijer, Chris J. L. M.; Middeldorp, Jaap M.

    1999-01-01

    A competitive quantitative PCR (Q-PCR) assay combined with simple silica-based DNA extraction was developed for monitoring of Epstein-Barr virus (EBV) DNA load in unfractionated peripheral blood. The Q-PCR is based on competitive coamplification of a highly conserved 213-bp region of the EBNA-1 open reading frame with an internal standard (IS), added in a known concentration. The IS has the same amplicon length and base composition as the wild-type (WT) EBNA-1 amplicon but differs in 23 internally randomized bases. Competitive coamplification yields two PCR products that are quantified by enzyme immunoassay or by electrochemiluminescence detection, with probes specific for the 23 differing internal nucleotides. The Q-PCR has a sensitivity of 10 copies of either WT or IS plasmid DNA. The Q-PCR was validated by quantification of known amounts of plasmid containing the WT EBNA-1 target. Furthermore, we determined EBV genome copy numbers in different cell lines. For EBV quantification in clinical samples, DNA was isolated from lysed whole blood by silica-affinity purification. Forty-six percent of healthy donor peripheral blood samples were positive by Q-PCR. In most of these samples, viral load was less than 2,000 EBV copies/ml of blood. In peripheral blood samples from two AIDS-related non-Hodgkin’s lymphoma patients, elevated EBV loads (up to 120,000 copies/ml) were observed, which decreased upon therapy. In Burkitt’s lymphoma patients, up to 4,592,000 EBV genome copies/ml of blood were detected. In conclusion, the EBNA-1-based Q-PCR assay provides a reproducible, accurate, and easy method for studying the relationship between EBV load and clinical parameters. PMID:10449464

  2. Transcriptomics analysis of lungs and peripheral blood of crystalline silica-exposed rats.

    PubMed

    Sellamuthu, Rajendran; Umbright, Christina; Roberts, Jenny R; Chapman, Rebecca; Young, Shih-Houng; Richardson, Diana; Cumpston, Jared; McKinney, Walter; Chen, Bean T; Frazer, David; Li, Shengqiao; Kashon, Michael; Joseph, Pius

    2012-08-01

    Minimally invasive approaches to detect/predict target organ toxicity have significant practical applications in occupational toxicology. The potential application of peripheral blood transcriptomics as a practical approach to study the mechanisms of silica-induced pulmonary toxicity was investigated. Rats were exposed by inhalation to crystalline silica (15 mg/m(3), 6 h/day, 5 days) and pulmonary toxicity and global gene expression profiles of lungs and peripheral blood were determined at 32 weeks following termination of exposure. A significant elevation in bronchoalveolar lavage fluid lactate dehydrogenase activity and moderate histological changes in the lungs, including type II pneumocyte hyperplasia and fibrosis, indicated pulmonary toxicity in the rats. Similarly, significant infiltration of neutrophils and elevated monocyte chemotactic protein-1 levels in the lungs showed pulmonary inflammation in the rats. Microarray analysis of global gene expression profiles identified significant differential expression [>1.5-fold change and false discovery rate (FDR) p < 0.01] of 520 and 537 genes, respectively, in the lungs and blood of the exposed rats. Bioinformatics analysis of the differentially expressed genes demonstrated significant similarity in the biological processes, molecular networks, and canonical pathways enriched by silica exposure in the lungs and blood of the rats. Several genes involved in functions relevant to silica-induced pulmonary toxicity such as inflammation, respiratory diseases, cancer, cellular movement, fibrosis, etc, were found significantly differentially expressed in the lungs and blood of the silica-exposed rats. The results of this study suggested the potential application of peripheral blood gene expression profiling as a toxicologically relevant and minimally invasive surrogate approach to study the mechanisms underlying silica-induced pulmonary toxicity.

  3. Advances towards reliable identification and concentration determination of rare cells in peripheral blood

    NASA Astrophysics Data System (ADS)

    Alemany Server, R.; Martens, D.; Jans, K.; Bienstman, P.; Hill, D.

    2016-03-01

    Through further development, integration and validation of micro-nano-bio and biophotonics systems FP7 CanDo is developing an instrument that will permit highly reproducible and reliable identification and concentration determination of rare cells in peripheral blood for two key societal challenges, early and low cost anti-cancer drug efficacy determination and cancer diagnosis/monitoring. A cellular link between the primary malignant tumour and the peripheral metastases, responsible for 90% of cancerrelated deaths, has been established in the form of circulating tumour cells (CTCs) in peripheral blood. Furthermore, the relatively short survival time of CTCs in peripheral blood means that their detection is indicative of tumour progression thereby providing in addition to a prognostic value an evaluation of therapeutic efficacy and early recognition of tumour progression in theranostics. In cancer patients however blood concentrations are very low (=1 CTC/1E9 cells) and current detection strategies are too insensitive, limiting use to prognosis of only those with advanced metastatic cancer. Similarly, problems occur in therapeutics with anti-cancer drug development leading to lengthy and costly trials often preventing access to market. The novel cell separation/Raman analysis technologies plus nucleic acid based molecular characterization of the CanDo platform will provide an accurate CTC count with high throughput and high yield meeting both key societal challenges. Being beyond the state of art it will lead to substantial share gains not just in the high end markets of drug discovery and cancer diagnostics but due to modular technologies also in others. Here we present preliminary DNA hybridization sensing results.

  4. [Surface architectonic of the peripheral red blood cells in cancer patients].

    PubMed

    Novitskiĭ, V V; Chasovskikh, N Iu; Stepovaia, E A; Gol'dberg, V E; Kitsmaniuk, Z D; Kolosova, M V; Mikhaĭlenko, A N; Koreshkova, K G; Bazhenova, N G; Petrova, I V; Sokolova, I B

    1997-08-01

    Scanning electron microscopy of the relief of the surface of peripheral red blood cells in patients with tumors of the head and neck (cancer of the tongue, larynx, and fundal buccal mucosa) of stages II-III and with stages III-IV lung cancer revealed similar changes, characterized by a manifest drop in the count of discocytes and increased share of transitional, prehemolytic, and degenerative forms of cells.

  5. Transcriptional Profiling of Francisella tularensis Infected Peripheral Blood Monomuclear Cells: A Predictive Tool for Tularemia

    DTIC Science & Technology

    2008-01-01

    responses (Munn et al., 1999). Genes associated with a role in lipid metabolism and PPAR pathways were also found to be mostly induced at a later time...infection, notably those involved in calcium, zinc ion binding, PPAR signaling, and lipid metabolism, which further refines the current knowledge of F...rate, may induce septicemia and fever without the primary involvement of lymph nodes or lungs (Ellis et al., 2002). Peripheral blood mononuclear cells

  6. Effect of amino acids on luminol-dependent chemiluminescence generated by peripheral blood polymorphonuclear leukocytes.

    PubMed Central

    Minuk, G Y; Rascanin, N; Woods, D E

    1986-01-01

    Individual amino acids and amino acid mixtures caused a dose-dependent increase in chemiluminescence generated by peripheral blood polymorphonuclear leukocytes. A visual assay for opsonophagocytosis, however, failed to identify any quantitative differences between leukocytes incubated with amino acids and those incubated in amino-acid-free solutions. The results of this study suggest that the presence of amino acids may interfere with the proper interpretation of luminol-dependent chemiluminescence curves. PMID:3745427

  7. Interleukin 1 production by peripheral blood mononuclear cells from leprosy patients.

    PubMed Central

    Watson, S; Bullock, W; Nelson, K; Schauf, V; Gelber, R; Jacobson, R

    1984-01-01

    Quantitation of interleukin 1 production by adherent mononuclear cells from peripheral blood was performed in patients with tuberculoid and lepromatous forms of leprosy. Cells from patients with tuberculoid leprosy either secreted interleukin 1 spontaneously or produced amounts within the normal range in response to lipopolysaccharide stimulation. Conversely, stimulated cells from lepromatous patients failed to produce interleukin 1 in 5 of 13 (38.5%) cases. PMID:6332077

  8. [Mobilization of peripheral blood stem cells with plerixafor in poor mobilizer patients].

    PubMed

    Sancho, Juan-Manuel; Duarte, Rafael; Medina, Laura; Querol, Sergi; Marín, Pedro; Sureda, Anna

    2016-09-02

    Poor mobilization of peripheral blood stem cells (CD34(+) cells) from bone marrow is a frequent reason for not reaching the autologous stem cell trasplantation (SCT) procedure in patients diagnosed with lymphoma or myeloma. Plerixafor, a reversible inhibitor of the binding of stromal cell-derived factor 1 to its cognate receptor CXCR4, has demonstrated a higher capacity for the mobilization of peripheral blood stem cells in combination with granulocyte colony stimulating factor (G-CSF) compared with G-CSF alone. For this reason, plerixafor is now indicated for poor mobilizer myeloma or lymphoma patients. Some studies have recently indicated that a pre-emptive strategy of plerixafor use during first mobilization, according to the number of CD34(+) mobilized cells in peripheral blood or to the harvested CD34(+) cells after first apheresis, could avoid mobilization failures and re-mobilizations, as well as the delay of autologous SCT. The aim of this consensus was to perform a review of published studies on pre-emptive strategy and to establish common recommendations for hospitals in Catalonia and Balearics on the use of pre-emptive plerixafor. For the Consensus, physicians from participant hospitals met to review previous studies as well as previous own data about plerixafor use. The GRADE system was used to qualify the available evidence and to establish recommendations on the use of pre-emptive plerixafor. After a review of the literature, the expert consensus recommended the administration of pre-emptive plerixafor for multiple myeloma or lymphoma patients with a CD34+ cell count lower than 10 cells/μL in peripheral blood (measured in the morning of day 4 of mobilization with G-CSF or after haematopietic recovery in the case of mobilization with chemotherapy plus G-CSF). Copyright © 2016 Elsevier España, S.L.U. All rights reserved.

  9. Small RNAs in the peripheral blood discriminate metastasized from non-metastasized seminoma

    PubMed Central

    2014-01-01

    Background We aimed to better discriminate metastasized (lymphogen/occult/both combined) from non-metastasized seminoma based on post-transcriptional changes examined in the peripheral blood. Methods Total RNAs including small RNAs were isolated from the peripheral blood of patients suffering from metastasized testicular tumours (lymphogen, n = 5, clinical stage IIb/c; occult, n = 5, clinical stage I) and non-metastasized patients (n = 5, clinical stage I). Small RNA next generation sequencing (SOLID, Life Technologies) was employed to examine post-transcriptional changes. We searched for small RNAs showing at least 50 reads and a significant ≥ 2-fold difference using peripheral blood small RNAs of non-metastasized tumours as the reference group. Candidate small RNAs were examined in univariate logistic regression analysis and combinations of two small RNAs were further examined using support vector machines. Results On average 1.3x107, 1.2x107 and 1.2x107 small RNA reads were detectable in non-metastasized, lymphogen and occult metastasized seminoma, respectively of which 73-76% remained after trimming. From these between 80-82% represented annotated reads and 7.2-7.8% (1.6-1.7x104) were annotated small RNA tags. Of them 137 small RNAs showed > 50 reads and a ≥ two-fold difference to the reference. In univariate analysis we detected 33-35 different small RNAs which significantly discriminated lymphogen/occult/combined metastasized from non-metastasized seminoma and among these different comparisons it were the same small RNAs in 44-79%. Many combinations of two of these small RNAs completely discriminated metastasized from non-metastasized seminoma irrespective of the metastasis subtype. Conclusions Metastasized (either lymphogen or occult) seminoma can be completely discriminated from non-metastasized seminoma with a combination of two small RNAs measured in the peripheral blood. PMID:24597607

  10. Small RNAs in the peripheral blood discriminate metastasized from non-metastasized seminoma.

    PubMed

    Ruf, Christian G; Dinger, Daniela; Port, Matthias; Schmelz, Hans-Ulrich; Wagner, Walter; Matthies, Cord; Müller-Myhsok, Bertram; Meineke, Viktor; Abend, Michael

    2014-03-06

    We aimed to better discriminate metastasized (lymphogen/occult/both combined) from non-metastasized seminoma based on post-transcriptional changes examined in the peripheral blood. Total RNAs including small RNAs were isolated from the peripheral blood of patients suffering from metastasized testicular tumours (lymphogen, n = 5, clinical stage IIb/c; occult, n = 5, clinical stage I) and non-metastasized patients (n = 5, clinical stage I). Small RNA next generation sequencing (SOLID, Life Technologies) was employed to examine post-transcriptional changes. We searched for small RNAs showing at least 50 reads and a significant  ≥ 2-fold difference using peripheral blood small RNAs of non-metastasized tumours as the reference group. Candidate small RNAs were examined in univariate logistic regression analysis and combinations of two small RNAs were further examined using support vector machines. On average 1.3 x 10(7), 1.2 x 10(7) and 1.2 x 10(7) small RNA reads were detectable in non-metastasized, lymphogen and occult metastasized seminoma, respectively of which 73-76% remained after trimming. From these between 80-82% represented annotated reads and 7.2-7.8% (1.6-1.7 x 10(4)) were annotated small RNA tags. Of them 137 small RNAs showed > 50 reads and a ≥ two-fold difference to the reference. In univariate analysis we detected 33-35 different small RNAs which significantly discriminated lymphogen/occult/combined metastasized from non-metastasized seminoma and among these different comparisons it were the same small RNAs in 44-79%. Many combinations of two of these small RNAs completely discriminated metastasized from non-metastasized seminoma irrespective of the metastasis subtype. Metastasized (either lymphogen or occult) seminoma can be completely discriminated from non-metastasized seminoma with a combination of two small RNAs measured in the peripheral blood.

  11. Prospective identification of neoantigen-specific lymphocytes in the peripheral blood of melanoma patients.

    PubMed

    Gros, Alena; Parkhurst, Maria R; Tran, Eric; Pasetto, Anna; Robbins, Paul F; Ilyas, Sadia; Prickett, Todd D; Gartner, Jared J; Crystal, Jessica S; Roberts, Ilana M; Trebska-McGowan, Kasia; Wunderlich, John R; Yang, James C; Rosenberg, Steven A

    2016-04-01

    Detection of lymphocytes that target tumor-specific mutant neoantigens--derived from products encoded by mutated genes in the tumor--is mostly limited to tumor-resident lymphocytes, but whether these lymphocytes often occur in the circulation is unclear. We recently reported that intratumoral expression of the programmed cell death 1 (PD-1) receptor can guide the identification of the patient-specific repertoire of tumor-reactive CD8(+) lymphocytes that reside in the tumor. In view of these findings, we investigated whether PD-1 expression on peripheral blood lymphocytes could be used as a biomarker to detect T cells that target neoantigens. By using a high-throughput personalized screening approach, we identified neoantigen-specific lymphocytes in the peripheral blood of three of four melanoma patients. Despite their low frequency in the circulation, we found that CD8(+)PD-1(+), but not CD8(+)PD-1(-), cell populations had lymphocytes that targeted 3, 3 and 1 unique, patient-specific neoantigens, respectively. We show that neoantigen-specific T cells and gene-engineered lymphocytes expressing neoantigen-specific T cell receptors (TCRs) isolated from peripheral blood recognized autologous tumors. Notably, the tumor-antigen specificities and TCR repertoires of the circulating and tumor-infiltrating CD8(+)PD-1(+) cells appeared similar, implying that the circulating CD8(+)PD-1(+) lymphocytes could provide a window into the tumor-resident antitumor lymphocytes. Thus, expression of PD-1 identifies a diverse and patient-specific antitumor T cell response in peripheral blood, providing a novel noninvasive strategy to develop personalized therapies using neoantigen-reactive lymphocytes or TCRs to treat cancer.

  12. Peripheral blood eosinophils: a surrogate marker for airway eosinophilia in stable COPD

    PubMed Central

    Negewo, Netsanet A; McDonald, Vanessa M; Baines, Katherine J; Wark, Peter AB; Simpson, Jodie L; Jones, Paul W; Gibson, Peter G

    2016-01-01

    Introduction Sputum eosinophilia occurs in approximately one-third of stable chronic obstructive pulmonary disease (COPD) patients and can predict exacerbation risk and response to corticosteroid treatments. Sputum induction, however, requires expertise, may not always be successful, and does not provide point-of-care results. Easily applicable diagnostic markers that can predict sputum eosinophilia in stable COPD patients have the potential to progress COPD management. This study investigated the correlation and predictive relationship between peripheral blood and sputum eosinophils. It also examined the repeatability of blood eosinophil counts. Methods Stable COPD patients (n=141) were classified as eosinophilic or noneosinophilic based on their sputum cell counts (≥3%), and a cross-sectional analysis was conducted comparing their demographics, clinical characteristics, and blood cell counts. Receiver operating characteristic curve analysis was used to assess the predictive ability of blood eosinophils for sputum eosinophilia. Intraclass correlation coefficient was used to examine the repeatability of blood eosinophil counts. Results Blood eosinophil counts were significantly higher in patients with sputum eosinophilia (n=45) compared to those without (0.3×109/L vs 0.15×109/L; P<0.0001). Blood eosinophils correlated with both the percentage (ρ=0.535; P<0.0001) and number of sputum eosinophils (ρ=0.473; P<0.0001). Absolute blood eosinophil count was predictive of sputum eosinophilia (area under the curve =0.76, 95% confidence interval [CI] =0.67–0.84; P<0.0001). At a threshold of ≥0.3×109/L (specificity =76%, sensitivity =60%, and positive likelihood ratio =2.5), peripheral blood eosinophil counts enabled identification of the presence or absence of sputum eosinophilia in 71% of the cases. A threshold of ≥0.4×109/L had similar classifying ability but better specificity (91.7%) and higher positive likelihood ratio (3.7). In contrast, ≥0.2×109/L

  13. Interaction of bovine peripheral blood polymorphonuclear cells and Leptospira species; innate responses in the natural bovine reservoir host.

    USDA-ARS?s Scientific Manuscript database

    Cattle are the reservoir hosts of Leptospira borgpetersenii serovar Hardjo, and also be reservoir hosts of other Leptospira species such as L. kirschneri, and L. interrogans. As a reservoir host, cattle shed Leptospira, infecting other animals, including humans. Previous studies with human and murin...

  14. Host-Based Peripheral Blood Gene Expression Analysis for Diagnosis of Infectious Diseases.

    PubMed

    Holcomb, Zachary E; Tsalik, Ephraim L; Woods, Christopher W; McClain, Micah T

    2017-02-01

    Emerging pandemic infectious threats, inappropriate antibacterial use contributing to multidrug resistance, and increased morbidity and mortality from diagnostic delays all contribute to a need for improved diagnostics in the field of infectious diseases. Historically, diagnosis of infectious diseases has relied on pathogen detection; however, a novel concept to improve diagnostics in infectious diseases relies instead on the detection of changes in patterns of gene expression in circulating white blood cells in response to infection. Alterations in peripheral blood gene expression in the infected state are robust and reproducible, yielding diagnostic and prognostic information to help facilitate patient treatment decisions.

  15. [Isolation of monocytes from peripheral blood by gradient centrifugation on Lymphoprep in a hypertonic medium].

    PubMed

    Májský, A

    1989-12-08

    The author describes a new method of isolation of monocytes from peripheral blood, involving hypertonization of the blood and isolation of monocytes by centrifuging above a gradient of hypertonic Lymphoprep. The method makes it possible to obtain a monocyte suspension with at least 85% cells; there are no B lymphocytes in the suspension and the monocytes are not damaged. The quality of the thus obtained monocytes was tested by examination with monocytic sera and with HLA-DR antibodies, whereby the test was concurrently made with monocytes obtained by adherence and with B lymphocytes.

  16. A Novel Needle-Free Blood Draw Device for Sample Collection From Short Peripheral Catheters

    PubMed Central

    Nachamkin, Irving

    2017-01-01

    A new US Food and Drug Administration-cleared needleless blood collection device (PIVO; Velano Vascular, San Francisco, CA) for short peripheral catheters was compared with conventional venipuncture for collecting blood samples for routine laboratory analysis from adult healthy volunteers. The PIVO device was comparable with venipuncture in terms of providing high-integrity samples (no hemolysis or clotting), equivalent laboratory values, and better patient experience as assessed by pain scores. Further studies to assess the overall utility of the PIVO device are warranted. PMID:28419012

  17. Adhesion of peripheral blood mononuclear cells to vascular endothelium in patients with systemic sclerosis (scleroderma).

    PubMed

    Rudnicka, L; Majewski, S; Blaszczyk, M; Skiendzielewska, A; Makiela, B; Skopinska, M; Jablonska, S

    1992-07-01

    Perivascular infiltrates in skin, subcutaneous tissue, and internal organs are a characteristic feature of early systemic sclerosis (SSc). We studied the first step of migration of peripheral blood mononuclear cells (PBMC) through the vessel wall to the extravascular space, i.e., adhesion of PBMC to endothelial cells (EC), in patients with various forms of SSc (limited scleroderma, diffuse scleroderma, and the transitional form). Radioisotope-labeled patient PBMC were coincubated with umbilical cord EC in vitro, and the percentage adhesion was measured. Adhesion of PBMC to EC was markedly decreased, while adhesion of isolated active rosette-forming cells (ARFC) was significantly increased, in SSc patients compared with healthy controls. Decreased adhesion of PBMC to EC was found to correlate with a diminished percentage of ARFC in the peripheral blood. Preincubation of PBMC from healthy donors with interleukin-2 (IL-2) enhanced their adhesion to EC, while preincubation of PBMC from SSc patients with this cytokine resulted in a decrease in adhesion in 10 of 14 individuals. IL-1, interferon-gamma, and transforming growth factor beta had no significant effect on adhesion of SSc patient PBMC to EC. Differences in adhesion of PBMC to EC among the SSc subgroups were not significant. Our findings suggest that in SSc, activation of subpopulations of PBMC leads to their enhanced adhesion to vascular endothelium in vivo and to migration of these cells to the extravascular space, resulting in the elimination from the peripheral blood of those PBMC with high ability to adhere to EC.

  18. Altered Immune Phenotype in Peripheral Blood Cells of Patients with Scleroderma-Associated Pulmonary Hypertension

    PubMed Central

    Risbano, Michael G; Meadows, Christina A; Coldren, Christopher D; Jenkins, Tiffany J.; Edwards, Michael G; Collier, David; Huber, Wendy; Mack, Douglas G; Fontenot, Andrew P; Geraci, Mark W; Bull, Todd M

    2010-01-01

    Pulmonary arterial hypertension is a common and fatal complication of scleroderma that may involve inflammatory and autoimmune mechanisms. Alterations in the gene expression of peripheral blood mononuclear cells have been previously described in patients with pulmonary arterial hypertension. Our goal is to identify differentially expressed genes in peripheral blood mononuclear cells in scleroderma patients with and without pulmonary hypertension as biomarkers of disease. Gene expression analysis was performed on a Microarray Cohort of scleroderma patients with (n=10) and without (n=10) pulmonary hypertension. Differentially expressed genes were confirmed in the Microarray Cohort and validated in a Validation Cohort of scleroderma patients with (n=15) and without (n=19) pulmonary hypertension by RT-qPCR. We identified inflammatory and immune-related genes including interleukin-7 receptor (IL-7R) and chemokine receptor 7 as differentially expressed in patients with scleroderma-associated pulmonary hypertension. Flow cytometry confirmed decreased expression of IL-7R on circulating CD4+ T-cells from scleroderma patients with pulmonary hypertension. Differences exist in the expression of inflammatory and immune-related genes in peripheral blood cells from patients with scleroderma-related pulmonary hypertension compared to those with normal pulmonary artery pressures. These findings may have implications as biomarkers to screen at-risk populations for early diagnosis and provide insight into mechanisms of scleroderma-related pulmonary hypertension. PMID:20973920

  19. Mutagenicity of the Musa paradisiaca (Musaceae) fruit peel extract in mouse peripheral blood cells in vivo.

    PubMed

    Andrade, C U B; Perazzo, F F; Maistro, E L

    2008-01-01

    Plants are a source of many biologically active products and nowadays they are of great interest to the pharmaceutical industry. In the present study, the mutagenic potential of the Musa paradisiaca fruit peel extract was assessed by the single-cell gel electrophoresis (SCGE) and micronucleus assays. Animals were treated orally with three different concentrations of the extract (1000, 1500, and 2000 mg/kg body weight). Peripheral blood cells of Swiss mice were collected 24 h after treatment for the SCGE assay and 48 and 72 h for the micronucleus test. The results showed that the two higher doses of the extract of M. paradisiaca induced statistically significant increases in the average numbers of DNA damage in peripheral blood leukocytes for the two higher doses and a significant increase in the mean of micronucleated polychromatic erythrocytes in the three doses tested. The polychromatic/normochromatic erythrocyte ratio scored in the treated groups was not statistically different from the negative control. The data obtained indicate that fruit peel extract from M. paradisiaca showed mutagenic effect in the peripheral blood cells of Swiss albino mice.

  20. Isolation and characterization of peripheral blood-derived endothelial progenitor cells from broiler chickens.

    PubMed

    Bi, Shicheng; Tan, Xun; Ali, Shah Qurban; Wei, Liangjun

    2014-11-01

    Peripheral blood-derived endothelial progenitor cells (EPCs) have been extensively studied in mammals but the isolation and characterization of EPCs in avian species have not been reported. In this study, chicken peripheral blood mononuclear cells (PBMNCs) were cultured under conditions favoring endothelial-specific differentiation for 2 weeks. One heterogeneous cell population dominated by spindle-shaped cells (early EPCs) and one homogeneous cell population exhibiting cobblestone-like morphology (endothelial outgrowth cells, EOCs) appeared sequentially. Quantitative polymerase chain reaction (PCR) showed the expression of several progenitor and endothelial cell markers such as CD133, VEGFR-2 and CD31 in both cell populations. However, CD34, another progenitor marker, was undetectable in either freshly isolated PBMNCs or cultured cells. The endothelial phenotype of the EOCs was further identified by acetylated low-density lipoprotein/lectin double staining, and in vitro tube formation. Collectively, these data demonstrate that chicken EPCs can be isolated and cultured from PBMNCs and suggest that EPCs obtained from peripheral blood may originate mainly from the CD34- subpopulation. Copyright © 2014 Elsevier Ltd. All rights reserved.

  1. Supernatant of Bone Marrow Mesenchymal Stromal Cells Induces Peripheral Blood Mononuclear Cells Possessing Mesenchymal Features

    PubMed Central

    Hu, Gang; Xu, Jun-jun; Deng, Zhi-hong; Feng, Jie; Jin, Yan

    2011-01-01

    Increasing evidence shows that some cells from peripheral blood fibroblast-like mononuclear cells have the capacity to differentiate into mesenchymal lineages. However, the insufficiency of these cells in the circulation challenges the cell isolation and subsequently limits the clinical application of these cells. In the present study, the peripheral blood mononuclear cells (pbMNCs) were isolated from wound animals and treated with the supernatant of bone marrow mesenchymal stromal cells (bmMSCs). Results showed these pbMNCs were fibroblast-like, had stromal morphology, were negative for CD34 and CD45, but positive for Vimentin and Collagen I, and had the multipotency to differentiate into adipocytes and osteoblasts. We named these induced peripheral blood-derived mesenchymal stromal cells (ipbMSCs). Skin grafts in combination with ipbMSCs and collagen I were applied for wound healing, and results revealed ipbMSC exhibited similar potency and effectiveness in the promotion of wound healing to the bmMSCs. Hereafter, we speculate that the mixture of growth factors and chemokines secreted by bmMSCs may play an important roles in the induction of the proliferation and mesenchymal differentiation of mononuclear cells. Our results are clinically relevant because it provide a new method for the acquisition of MSCs which can be used as a candidate for the wound repair. PMID:21494428

  2. IL6 Signaling in Peripheral Blood T Cells Predicts Clinical Outcome in Breast Cancer.

    PubMed

    Wang, Lei; Miyahira, Andrea K; Simons, Diana L; Lu, Xuyang; Chang, Andrew Y; Wang, Carrie; Suni, Maria A; Maino, Vernon C; Dirbas, Frederick M; Yim, John; Waisman, James; Lee, Peter P

    2017-03-01

    IL6 is a pleiotropic cytokine with both pro- and anti-inflammatory properties, which acts directly on cancer cells to promote their survival and proliferation. Elevated serum IL6 levels negatively correlate with survival of cancer patients, which is generally attributed to the direct effects of IL6 on cancer cells. How IL6 modulates the host immune response in cancer patients is unclear. Here, we show the IL6 signaling response in peripheral blood T cells is impaired in breast cancer patients and is associated with blunted Th17 differentiation. The mechanism identified involved downregulation of gp130 and IL6Rα in breast cancer patients and was independent of plasma IL6 levels. Importantly, defective IL6 signaling in peripheral blood T cells at diagnosis correlated with worse relapse-free survival. These results indicate that intact IL6 signaling in T cells is important for controlling cancer progression. Furthermore, they highlight a potential for IL6 signaling response in peripheral blood T cells at diagnosis as a predictive biomarker for clinical outcome of breast cancer patients. Cancer Res; 77(5); 1119-26. ©2016 AACR. ©2016 American Association for Cancer Research.

  3. Reduced Numbers and Impaired Function of Regulatory T Cells in Peripheral Blood of Ischemic Stroke Patients

    PubMed Central

    Ruhnau, Johanna; Schulze, Juliane; von Sarnowski, Bettina; Heinrich, Marie; Langner, Sönke; Wilden, Anika; Kessler, Christof; Bröker, Barbara M.

    2016-01-01

    Background and Purpose. Regulatory T cells (Tregs) have been suggested to modulate stroke-induced immune responses. However, analyses of Tregs in patients and in experimental stroke have yielded contradictory findings. We performed the current study to assess the regulation and function of Tregs in peripheral blood of stroke patients. Age dependent expression of CD39 on Tregs was quantified in mice and men. Methods. Total FoxP3+ Tregs and CD39+FoxP3+ Tregs were quantified by flow cytometry in controls and stroke patients on admission and on days 1, 3, 5, and 7 thereafter. Treg function was assessed by quantifying the inhibition of activation-induced expression of CD69 and CD154 on T effector cells (Teffs). Results. Total Tregs accounted for 5.0% of CD4+ T cells in controls and <2.8% in stroke patients on admission. They remained below control values until day 7. CD39+ Tregs were most strongly reduced in stroke patients. On day 3 the Treg-mediated inhibition of CD154 upregulation on CD4+ Teff was impaired in stroke patients. CD39 expression on Treg increased with age in peripheral blood of mice and men. Conclusion. We demonstrate a loss of active FoxP3+CD39+ Tregs from stroke patient's peripheral blood. The suppressive Treg function of remaining Tregs is impaired after stroke. PMID:27073295

  4. Mitochondrial Alterations in Peripheral Mononuclear Blood Cells from Alzheimer's Disease and Mild Cognitive Impairment Patients.

    PubMed

    Delbarba, A; Abate, G; Prandelli, C; Marziano, M; Buizza, L; Arce Varas, N; Novelli, A; Cuetos, F; Martinez, C; Lanni, C; Memo, M; Uberti, D

    2016-01-01

    It is well recognized that mitochondrial dysfunction contributes to neurodegeneration occurring in Alzheimer's disease (AD). However, evidences of mitochondrial defects in AD peripheral cells are still inconclusive. Here, some mitochondrial-encoded and nuclear-encoded proteins, involved in maintaining the correct mitochondria machine, were investigated in terms of protein expression and enzymatic activity in peripheral blood mononuclear cells (PBMCs) isolated from AD and Mild Cognitive Impairment (MCI) patients and healthy subjects. In addition mitochondrial DNA copy number was measured by real time PCR. We found some differences and some similarities between AD and MCI patients when compared with healthy subjects. For example, cytochrome C and cytochrome B were decreased in AD, while MCI showed only a statistical reduction of cytochrome C. On the other hand, both AD and MCI blood cells exhibited highly nitrated MnSOD, index of a prooxidant environment inside the mitochondria. TFAM, a regulator of mitochondrial genome replication and transcription, was decreased in both AD and MCI patients' blood cells. Moreover also the mitochondrial DNA amount was reduced in PBMCs from both patient groups. In conclusion these data confirmed peripheral mitochondria impairment in AD and demonstrated that TFAM and mtDNA amount reduction could be two features of early events occurring in AD pathogenesis.

  5. Comparative Transcriptome Analysis Reveals Early Pregnancy-Specific Genes Expressed in Peripheral Blood of Pregnant Sows

    PubMed Central

    Zhu, Shien; Shi, Wenqing; Hu, Maishun; Fu, Xiangwei; Wang, Chuduan; Wang, Yachun; Zhang, Qin; Yu, Ying

    2014-01-01

    Early and accurate diagnosis of pregnancy is important for effective management of an economical pig farm. Besides the currently available methods used in early diagnosis of sows, circulating nucleic acids in peripheral blood may contain some early pregnancy-specific molecular markers. For the first time, microarray analysis of peripheral blood from pregnant sows versus non-pregnant sows identified 127 up-regulated and 56 down-regulated genes at day 14 post-insemination. Gene Ontology annotation grouped the total differently expressed genes into 3 significantly enriched terms, cell surface receptor linked signal transduction, G-protein coupled receptor protein signaling pathway and regulation of vesicle-mediated transport. Signaling pathway analysis revealed the only one significantly changed pathway was arachidonic acid metabolism. Of the differently expressed genes, nine (including LPAR3, RXFP4, GALP, CBR1, CBR2, GPX6, USP18, LHB and NR5A1) were found to exert function related to early pregnancy processes. This study provides a clue that differentially abundant RNAs in maternal peripheral blood can help to identify the molecular markers of early pregnancy in pigs. PMID:25479131

  6. Fever after peripheral blood stem cell infusion in haploidentical transplantation with post-transplant cyclophosphamide.

    PubMed

    Arango, Marcos; Combariza, Juan F

    2017-06-01

    Noninfection-related fever can occur after peripheral blood stem cell infusion in haploidentical hematopoietic stem cell transplantation with post-transplant cyclophosphamide. The objective of this study was to analyze the incidence of fever and characterize some clinical features of affected patients. A retrospective case-series study with 40 patients who received haploidentical hematopoietic stem cell transplantation was carried out. Thirty-three patients (82.5%) developed fever; no baseline characteristic was associated with its development. Median time to fever onset was 25.5h (range, 9.5-100h) and median peak temperature was 39.0°C (range, 38.1-40.5°C). Not a single patient developed hemodynamic or respiratory compromise that required admission to the intensive care unit. Fever was not explained by infection in any case. Ninety-one percent of the febrile episodes resolved within 96h of cyclophosphamide administration. No significant difference in overall survival, event-free survival, or graft versus host disease-free/relapse-free survival was found in the group of febrile individuals after peripheral blood stem cell infusion. Fever after peripheral blood stem cell infusion in this clinical setting was common; it usually subsides with cyclophosphamide administration. The development of fever was not associated with an adverse prognosis. Copyright © 2017 King Faisal Specialist Hospital & Research Centre. Published by Elsevier B.V. All rights reserved.

  7. Bone marrow and peripheral blood globin chain synthesis in sickle cell beta zero thalassaemia.

    PubMed Central

    Costa, F F; Zago, M A

    1986-01-01

    A similar imbalance of globin chain synthesis, with low non-alpha/alpha ratios, was shown in peripheral blood and in bone marrow of compound heterozygotes for both the Hb S and beta zero thalassaemia genes (S/beta zero thalassaemia). Previous purification of whole cell globin obtained from the bone marrow did not change the non-alpha/alpha ratio. The mean non-alpha/alpha ratios were 0.57 +/- 0.13 (means +/- SD) for the peripheral blood of 12 patients, 0.52 +/- 0.10 for five patients using bone marrow globin purified on Sephadex G100, and 0.55 +/- 0.16 for the unfiltered bone marrow globin of five patients. The data show that patients with S/beta zero thalassaemia have a similar beta chain deficiency in reticulocytes and in bone marrow cells, provided whole cell globin is used which avoids the removal of the free alpha chains. The non-alpha/alpha ratios in the peripheral blood of an S/beta zero thalassaemia patient and a beta thalassaemia heterozygote from the same family were compared in seven families and no significant difference was found. PMID:3723554

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

    SciTech Connect

    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.

  9. Seasonal variation of peripheral blood leukocyte telomere length in Costa Rica: a population based observational study

    PubMed Central

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

    2014-01-01

    Objectives Peripheral blood leukocyte telomere length is increasingly being used as a biomarker of aging, but its natural variation in human populations is not well understood. Several other biomarkers show seasonal variation, as do several determinants of leukocyte telomere length. We examined whether there was monthly variation in leukocyte telomere length in Costa Rica, a country with strong seasonal differences in precipitation and infection. Methods We examined a longitudinal population based cohort of 581 Costa Rican adults age 60 and above, from which blood samples were drawn between October 2006 and July 2008. Leukocyte telomere length was assayed from these samples using the quantitative PCR method. Multivariate regression models were used to examine correlations between month of blood draw and leukocyte telomere length. Results Telomere length from peripheral blood leukocytes varied by as much as 200 base pairs depending on month of blood draw, and this difference is not likely to be due to random variation. A moderate proportion of this association is statistically accounted for by month and region specific average rainfall. We found shorter telomere length associated with greater rainfall. Conclusions There are two possible explanations of our findings. First, there could be relatively rapid month-to-month changes in leukocyte telomere length. This conclusion would have implications for understanding the natural population dynamics of telomere length. Second, there could be seasonal differences in constituent cell populations. This conclusion would suggest that future studies of leukocyte telomere length use methods to account for the potential impact of constituent cell type. PMID:24615938

  10. Identification of characteristic molecular signature for volatile organic compounds in peripheral blood of rat

    SciTech Connect

    Kim, Jeong Kyu; Jung, Kwang Hwa; Noh, Ji Heon; Eun, Jung Woo; Bae, Hyun Jin; Xie, Hong Jian; Jang, Ja-June; Ryu, Jae Chun; Park, Won Sang; Lee, Jung Young; Nam, Suk Woo

    2011-01-15

    In a previous report we demonstrated that the transcriptomic response of liver tissue was specific to toxicants, and a characteristic molecular signature could be used as an early prognostic biomarker in rats. It is necessary to determine the transcriptomic response to toxicants in peripheral blood for application to the human system. Volatile organic compounds (VOCs) comprise a major group of pollutants which significantly affect the chemistry of the atmosphere and human health. In this study we identified and validated the specific molecular signatures of toxicants in rat whole blood as early predictors of environmental toxicants. VOCs (dichloromethane, ethylbenzene, and trichloroethylene) were administered to 11-week-old SD male rats after 48 h of exposure, peripheral whole blood was subjected to expression profiling analysis. Unsupervised gene expression analysis resulted in a characteristic molecular signature for each toxicant, and supervised analysis identified 1,217 outlier genes as distinct molecular signatures discerning VOC exposure from healthy controls. Further analysis of multi-classification suggested 337 genes as early detective molecular markers for three VOCs with 100% accuracy. A large-scale gene expression analysis of a different VOC exposure animal model suggested that characteristic expression profiles exist in blood cells and multi-classification of this VOC-specific molecular signature can discriminate each toxicant at an early exposure time. This blood expression signature can thus be used as discernable surrogate marker for detection of biological responses to VOC exposure in an environment.

  11. Peripheral arterial blood pressure versus central crterial blood pressure monitoring in critically ill patients after Cardio-pulmonary Bypass.

    PubMed

    Ahmad, Rana Altaf; Ahmad, Suhail; Naveed, Anjum; Baig, Mirza Ahmad Raza

    2017-01-01

    To determine the accuracy of peripheral (radial) arterial access as compared to central (femoral) arterial access for measurement of invasive blood pressure (IBP) in critically ill patients after cardiopulmonary bypass. Sixty patients (60) who required high inotropic/vasopressor support on weaning from cardio-pulmonary bypass and weaned off in 2(nd) attempt were included in this study. The duration of this study was from June 2015 to August 2016. Radial and femoral arterial access was achieved in all patients for simultaneous measurement of blood pressure. Arterial pressures were noted after 5, 15 and 30 minutes of weaning from cardiopulmonary bypass for both radial and femoral artery simultaneously. Mean age of study patients was 56.48±11.17 years. 85% patients were male. There was significant difference in systolic blood pressure, diastolic blood pressure and mean arterial pressures between the radial artery and femoral artery cannulation. Mean arterial pressures were significantly high in femoral artery as compared to the radial artery. The mean arterial pressures after five minutes of weaning using central access were 76.28±10.21 mmHg versus 64.15±6.76 mmHg in peripheral arterial access (p-value <0.001). Similarly we also found significant difference in mean arterial pressures after 15 minutes of weaning from cardiopulmonary bypass 78.70±10.12 mmHg in central access versus 72.03±6.76 mmHg using peripheral arterial access (p-value <0.001). The difference in arterial pressures were less marked as compared to the previous differences after 30 minutes of weaning from cardiopulmonary bypass as compared to the earlier readings (p-value 0.001). Peripheral arterial pressures are unreliable in critically ill patients after cardiopulmonary bypass receiving high dose of inotropic drugs. Central arterial access should be used in these patients to get accurate estimates of patients' blood pressure in early periods after cardiopulmonary bypass.

  12. Comparative study of peripheral blood smear and quantitative buffy coat in malaria diagnosis.

    PubMed

    Salmani, Manjunath P; Preeti, B Mindolli; Peerapur, B V

    2011-03-01

    A rapid test for diagnosis of malaria based on acridine orange staining of centrifuged blood samples in a microhaematocrit tube (QBC) was compared with Leishman stained thin peripheral blood smear in 287 samples. Malaria was diagnosed in 44 patients by Leishman staining technique and in 65 patients by QBC method. The QBC method allowed detection of an additional 21 cases. Thus the prevalence rate of malaria during the study was 22.65%. In 222 Patients who were negative by the QBC technique, the Leishman stained smears were also negative for malarial parasite. Although QBC method was superior to the smear for malarial parasite detection, species identification was difficult by this technique. The QBC method provides a reliable, quick, easily mastered, accurate method for diagnosis of malaria. The QBC system can also be used in the diagnosis of other parasitic diseases from blood (Filariasis). However, Leishman stained thin blood film still appear superior for species identification.

  13. Association of bovine Toll-like receptor 4 with tick infestation rates and blood histamine concentration.

    PubMed

    Zhao, G; Yu, M; Cui, Q-W; Zhou, X; Zhang, J-C; Li, H-X; Qu, K-X; Wang, G-L; Huang, B-Z

    2013-02-28

    We investigated a possible association between bovine Toll-like receptor 4 (TLR4) and resistance to tick infestation in 103 cattle, including BMY cattle (1/2 Brahman, 1/4 Murray Grey, and 1/4 Yunnan Yellow cattle), Brahman, and Red Angus grazing on improved pasture. The tick infestation weight and number of Rhipicephalus microplus and the blood histamine concentration were measured and compared with those of 32 Chinese Holsteins and 30 Simmentals. A 228-bp fragment was amplified and sequenced to analyze the polymorphisms of the TLR4 gene. After SSCP and sequencing analysis, 4 SNPs, i.e., 535(A>C), 546(T>C), 605(T>A), and 618(G>C), were identified, corresponding to GenBank accession Nos. AY297041 and NW_003104150; the latter two SNPs caused Leu→Gln and Gln→His substitutions, respectively. Genotype AA was completely predominant in the Chinese Holstein and Simmental; genotypes AA and AB were detected in Red Angus, while genotypes AA, AB, BB, and BC were detected in Brahman and in BMY cattle. A negative correlation was identified between blood histamine concentration and number of tick infestation; in BMY cattle this negative association was significant. The tick infestation in cattle with genotype BB was significantly lower than in those with genotype AA. Blood histamine concentration in cattle with genotype BB was significantly higher than in those with genotype AA. The TLR4 gene mutation could affect the blood histamine level and activate the immune reaction after tick infestation. Allele B has potential as a molecular marker for tick-resistance originated from Zebu cattle for use in cattle breeding programs.

  14. Gene expression analysis of whole blood, peripheral blood mononuclear cells, and lymphoblastoid cell lines from the Framingham Heart Study

    PubMed Central

    Joehanes, Roby; Johnson, Andrew D.; Barb, Jennifer J.; Raghavachari, Nalini; Liu, Poching; Woodhouse, Kimberly A.; O'Donnell, Christopher J.; Munson, Peter J.

    2012-01-01

    Despite a growing number of reports of gene expression analysis from blood-derived RNA sources, there have been few systematic comparisons of various RNA sources in transcriptomic analysis or for biomarker discovery in the context of cardiovascular disease (CVD). As a pilot study of the Systems Approach to Biomarker Research (SABRe) in CVD Initiative, this investigation used Affymetrix Exon arrays to characterize gene expression of three blood-derived RNA sources: lymphoblastoid cell lines (LCL), whole blood using PAXgene tubes (PAX), and peripheral blood mononuclear cells (PBMC). Their performance was compared in relation to identifying transcript associations with sex and CVD risk factors, such as age, high-density lipoprotein, and smoking status, and the differential blood cell count. We also identified a set of exons that vary substantially between participants, but consistently in each RNA source. Such exons are thus stable phenotypes of the participant and may potentially become useful fingerprinting biomarkers. In agreement with previous studies, we found that each of the RNA sources is distinct. Unlike PAX and PBMC, LCL gene expression showed little association with the differential blood count. LCL, however, was able to detect two genes related to smoking status. PAX and PBMC identified Y-chromosome probe sets similarly and slightly better than LCL. PMID:22045913

  15. A functional comparison of IIIindium-labelled elicited peripheral blood neutrophils and peritoneal neutrophils in the rat.

    PubMed Central

    Savige, J A; Saverymuttu, S H; Pinching, A J

    1984-01-01

    A functional comparison between elicited peripheral blood neutrophils has been made in vivo and in vitro. Preliminary experiments showed that separation of peripheral blood cells on a metrizamide gradient yielded too few neutrophils for efficient radiolabelling with indium (In): hence a mixed cell preparation comprising 80% neutrophils was elicited in the peripheral blood of adult male rats by the administration of endotoxin (0.25 mg i.a.) and cobra venom factor (200 microliter i.p.) 20 h before. Peritoneal neutrophils were collected 4 h after the i.p. injection of 6 ml thioglycollate. Both populations differed markedly from normal peripheral neutrophils on the in vitro testing of random locomotion, chemotaxis and phagocytosis of Candida. After labelling with IIIIn-tropolonate, a greater proportion (mean = 8%) of peripheral blood cells localized to an E. coli/Freund's complete adjuvant-induced abscess compared with peritoneal neutrophils (mean = 3%). The abscess could be visualized externally by scanning with both cell preparations, but the distribution of activity differed markedly. The greater hepatic sequestration of peritoneal neutrophils suggested cell damage or activation. To overcome the difficulty of harvesting normal peripheral blood neutrophils in the rat, either of these populations can be used to follow the kinetics of inflammation. However, elicited peripheral blood cells yield a higher proportion of responding cells. Images p740-a Fig. 1 PMID:6509802

  16. Peripheral Blood Stem Cell Transplant Related Plasmodium falciparum Infection in a Patient with Sickle Cell Disease

    PubMed Central

    Mejia, Rojelio; Booth, Garrett S.; Fedorko, Daniel P.; Hsieh, Matthew M.; Khuu, Hanh M.; Klein, Harvey G.; Mu, Jianbing; Fahle, Gary; Nutman, Thomas B.; Su, Xin-Zhuan; Williams, Esther C.; Flegel, Willy A.; Klion, Amy

    2012-01-01

    Background Although transmission of Plasmodium falciparum (Pf) infection during red blood cell transfusion from an infected donor has been well documented, malaria parasites are not known to infect hematopoietic stem cells. We report a case of Pf infection in a patient 11 days after peripheral blood stem cell transplant for sickle cell disease. Study Design and Methods Malaria parasites were detected in thick blood smears by Giemsa staining. Pf HRP2 antigen was measured by ELISA on whole blood and plasma. Pf DNA was detected in whole blood and stem cell retention samples by real-time PCR using Pf species–specific primers and probes. Genotyping of 8 Pf microsatellites was performed on genomic DNA extracted from whole blood. Results Pf was not detected by molecular, serologic or parasitologic means in samples from the recipient until day 11 post-transplant, coincident with the onset of symptoms. In contrast, Pf antigen was retrospectively detected in stored plasma collected 3 months prior to transplant from the asymptomatic donor. Pf DNA was detected in whole blood from both the donor and recipient post-transplant, and genotyping confirmed shared markers between donor and recipient Pf strains. Look back analysis of red blood cell donors was negative for Pf infection. Conclusions These findings are consistent with transmission by the stem cell product and have profound implications with respect to the screening of potential stem cell donors and recipients from malaria-endemic regions. PMID:22536941

  17. Does pulmonary rehabilitation reduce peripheral blood pressure in patients with chronic obstructive pulmonary disease?

    PubMed

    Canavan, Jane L; Kaliaraju, Djeya; Nolan, Claire M; Clark, Amy L; Jones, Sarah E; Kon, Samantha S C; Polkey, Michael I; Man, William D-C

    2015-08-01

    Pulmonary rehabilitation (PR) can improve aerobic exercise capacity, health-related quality of life and dyspnoea in patients with chronic obstructive pulmonary disease (COPD). Recent studies have suggested that exercise training may improve blood pressure and arterial stiffness, albeit in small highly selected cohorts. The aim of the study was to establish whether supervised outpatient or unsupervised home PR can reduce peripheral blood pressure. Resting blood pressure was measured in 418 patients with COPD before and after outpatient PR, supervised by a hospital-based team (HOSP). Seventy-four patients with COPD undergoing an unsupervised home-based programme acted as a comparator group (HOME). Despite significant improvements in mean (95% confidence interval) exercise capacity in the HOSP group (56 (50-60) m, p < 0.001) and HOME group (30 (17-42) m, p < 0.001) systolic blood pressure (SBP), diastolic blood pressure (DBP) and mean arterial blood pressure (MAP) did not change in either the HOSP (SBP: p = 0.47; DBP: p = 0.06; MAP: p = 0.38) or HOME group (SBP: p = 0.67; DBP: p = 0.38; MAP: p = 0.76). Planned subgroup analysis of HOSP patients with known hypertension and/or cardiovascular disease showed no impact of PR upon blood pressure. PR is unlikely to reduce blood pressure, and by implication, makes a mechanism of action in which arterial stiffness is reduced, less likely.

  18. Bovine central memory T cells are highly proliferative in response to bovine tuberculosis infection

    USDA-ARS?s Scientific Manuscript database

    Long-term (i.e., 14 days) cultured IFN-gamma responses of peripheral blood mononuclear cells are used as a correlate of T cell central memory (Tcm) responses in both humans and cattle. With bovine tuberculosis, vaccine-elicited long-term IFN-gamma ELISPOT assays are a correlate of protection. Recent...

  19. The relationship between complete blood count parameters and Fontaine's Stages in patients with peripheral arterial disease.

    PubMed

    Demirtas, Sinan; Karahan, Oguz; Yazici, Suleyman; Guclu, Orkut; Caliskan, Ahmet; Yavuz, Celal; Kucuker, Aslihan; Mavitas, Binali

    2014-12-01

    The aim of the present study is to evaluate whether blood count parameters differ according to the stages of Fontaine's classification and to investigate the relationship between hemogram parameters and the severity of the disease. Eighty-two peripheral arterial disease patients were examined prospectively. Patients were classified according to the Fontaine classification system. Fifty newly diagnosed patients were included in the study. The neutrophil-to-lymphocyte ratio, mean platelet volume, and red blood cell distribution width values were recorded. Mean neutrophil-to-lymphocyte ratio values were found to be 3.31 ± 1.1% in Stage I, 3.11 ± 1.3% in Stage II, and 3.48 ± 1.1% in Stage III (p > 0.05). Mean platelet volume values were found to be 7.8 ± 0.6 fl (Stage I), 8.2 ± 1.0 fl (Stage II), and 9.0 ± 0.9 fl (Stage III) (p < 0.05). Red blood cell distribution width values were found to be 13.6 ± 1.0% in Stage I, 14.8 ± 1.7% in Stage II, and 15.4 ± 2.3% in Stage III, being significantly different among all three stages (p < 0.05). Both red blood cell distribution width and mean platelet volume are found to be associated with the severity of atherosclerotic disease in patients with peripheral arterial disease. This finding hypothesizes that complete blood counting parameters may serve as a beneficial and cost-effective method for monitoring atherosclerotic peripheral disease. © The Author(s) 2014 Reprints and permissions: sagepub.co.uk/journalsPermissions.nav.

  20. Peripheral blood metabolic and inflammatory factors as biomarkers to ocular findings in diabetic macular edema

    PubMed Central

    Sala-Puigdollers, Anna; Matas, Jessica; Vinagre, Irene; Ríos, José; Adán, Alfredo

    2017-01-01

    Aims To study the association between peripheral blood metabolic and inflammatory factors and presence of diabetic macular edema (DME) and its related anatomic features in type 2 diabetic mellitus (T2DM) patients. Material and methods Observational cross-sectional study on a proof of concept basis. Seventy-six T2DM included patients were divided based on the presence (n = 58) or absence of DME (n = 18) according to optical coherence tomography (OCT). Ultra-widefield fluorescein angiography (UWFA) was performed in DME patients. Fasting peripheral blood sample testing included glycemia, glycated hemoglobin, creatinin and lipid levels among others. Serum levels of a broad panel of cytokines and inflammatory mediators were also analysed. OCT findings included central subfoveal thickness, diffuse retinal thickness (DRT), cystoid macular edema (CME), serous retinal detachment and epirretinal membrane. UWFA items included pattern of DME, presence of peripheral retinal ischemia and enlarged foveal avascular zone (FAZ). Results Metabolic and inflammatory factors did not statistically differ between groups. However, several inflammatory mediators did associate to certain ocular items of DME cases: IL-6 was significantly higher in patients with DRT (p = 0.044), IL-10 was decreased in patients with CME (p = 0.012), and higher IL-8 (p = 0.031) and VEGF levels (p = 0.031) were observed in patients with enlarged FAZ. Conclusion Inflammatory and metabolic peripheral blood factors in T2DM may not be differentially associated to DME when compared to non-DME cases. However, some OCT and UWFA features of DME such as DRT, CME and enlarged FAZ may be associated to certain systemic inflammatory mediators. PMID:28328965

  1. Peripheral blood metabolic and inflammatory factors as biomarkers to ocular findings in diabetic macular edema.

    PubMed

    Figueras-Roca, Marc; Molins, Blanca; Sala-Puigdollers, Anna; Matas, Jessica; Vinagre, Irene; Ríos, José; Adán, Alfredo

    2017-01-01

    To study the association between peripheral blood metabolic and inflammatory factors and presence of diabetic macular edema (DME) and its related anatomic features in type 2 diabetic mellitus (T2DM) patients. Observational cross-sectional study on a proof of concept basis. Seventy-six T2DM included patients were divided based on the presence (n = 58) or absence of DME (n = 18) according to optical coherence tomography (OCT). Ultra-widefield fluorescein angiography (UWFA) was performed in DME patients. Fasting peripheral blood sample testing included glycemia, glycated hemoglobin, creatinin and lipid levels among others. Serum levels of a broad panel of cytokines and inflammatory mediators were also analysed. OCT findings included central subfoveal thickness, diffuse retinal thickness (DRT), cystoid macular edema (CME), serous retinal detachment and epirretinal membrane. UWFA items included pattern of DME, presence of peripheral retinal ischemia and enlarged foveal avascular zone (FAZ). Metabolic and inflammatory factors did not statistically differ between groups. However, several inflammatory mediators did associate to certain ocular items of DME cases: IL-6 was significantly higher in patients with DRT (p = 0.044), IL-10 was decreased in patients with CME (p = 0.012), and higher IL-8 (p = 0.031) and VEGF levels (p = 0.031) were observed in patients with enlarged FAZ. Inflammatory and metabolic peripheral blood factors in T2DM may not be differentially associated to DME when compared to non-DME cases. However, some OCT and UWFA features of DME such as DRT, CME and enlarged FAZ may be associated to certain systemic inflammatory mediators.

  2. Evaluation of new automated hematopoietic progenitor cell analysis in the clinical management of peripheral blood stem cell collections

    PubMed Central

    Peerschke, Ellinor I.; Moung, Christine; Pessin, Melissa S.; Maslak, Peter

    2016-01-01

    BACKGROUND Successful peripheral blood stem cell transplantation (PBSCT) depends on the collection and infusion of adequate numbers of peripheral blood progenitor cells (PBPCs). Several predictors of PBPC yield are used currently, including white blood cell (WBC) count and CD34 analysis. This study evaluated the utility of the new automated hematopoietic progenitor cell count available on Sysmex XN hematology analyzers (XN-HPCs) in PBSCT. STUDY DESIGN AND METHODS The performance characteristics of XN-HPC, CD34+, and WBC analysis were compared using 107 matched peripheral blood and apheresis samples. RESULTS Good correlation was observed between XN-HPC and CD34+ cell counts in peripheral blood (r = 0.88; slope, 0.81) and apheresis collections (r = 0.91; slope, 0.89). Moreover, peripheral blood XN-HPC and CD34 analysis showed comparable ability to predict successful PBPC harvests (≥ 2 × 106 CD34+ cells/kg). At a cutoff of 20 × 106 progenitor cells/L, peripheral blood XN- HPC and CD34 analysis both showed negative predictive values (NPVs) of 100% and positive predictive values (PPVs) of 55.4 and 63%, respectively. Using an optimized cutoff of 38 × 106 progenitor cells/L, derived from receiver operating characteristic analysis, the PPV for XN-HPC and CD34 analysis increased to 71.4 and 78.9%, respectively, with relatively unchanged NPVs (XN-HPC 97.7%, CD34+ 98.0%). In contrast, the correlation between peripheral blood WBC and CD34 analysis was poor (r = 0.48; slope, 669.85), and the peripheral blood WBC count (cutoff, 10 × 109/L) was a poor predictor of PBPC harvest (NPV 60%, PPV 43.1%). CONCLUSION XN-HPC compares favorably with CD34 analysis and may be a surrogate for CD34 analysis to predict optimal timing of PBPC collections. PMID:25808236

  3. [Expression of elafin in peripheral blood in inflammatory bowel disease patients and its clinical significance].

    PubMed

    Zhang, W; Teng, G G; Tian, Y; Wang, H H

    2016-04-12

    To quantify the expression of elafin mRNA in peripheral blood in patients with inflammatory bowel disease (IBD) and to explore its value in assessment of the activity and severity of IBD. From July 1 2015 to August 15 2015, 23 patients with IBD admitted to Peking University First Hospital were selected, including 15 cases with ulcerative colitis (UC) and 8 cases with Crohn's disease (CD). Among those, 5 cases were in remission (UC 3, CD 2), 6 cases were mild active (UC 3, CD 3), 3 cases were moderate active (UC 1, CD 2), and 9 cases were severe active (UC 8, CD 1). A total of 21 healthy individuals were selected as the control group. Peripheral blood samples of IBD patients and healthy controls were collected. The expression of elafin mRNA in peripheral blood leukocytes was detected by fluorescence quantitative real-time PCR. Mann-Whitney test was performed for comparison between the two groups. The correlation between the expression of elafin mRNA in peripheral blood and IBD activity score was analyzed by Pearson correlation analysis after transformation of variables. The median expression of elafin mRNA in peripheral blood leukocytes in IBD group and control group was 0.005 8 (0.000 2, 0.043 5) and 0.015 3 (0.002 1, 0.175 8), respectively, with no significant difference (P>0.05). However, in the active IBD patients it was lower than that in the controls (0.004 6 (0.000 2, 0.034 8) vs 0.015 3 (0.002 1, 0.175 8), P<0.05) and also lower than that in the remission patients(0.004 6 (0.000 2, 0.034 8) vs 0.023 1 (0.012 6, 0.043 5), P<0.05); in the active UC patients it was lower than that in the controls(0.003 7 (0.000 2, 0.027 0) vs 0.015 3 (0.002 1, 0.175 8), P<0.05). The expression of elafin mRNA in peripheral blood was negatively correlated with modified Mayo score in UC patients (r=-0.513, P<0.05) and with the Crohn's Disease Activity Index (CDAI) of Best score in CD patients (r=-0.889, P<0.05). The expression of elafin mRNA in peripheral blood in active IBD

  4. Comparison between tsetse fly synthetic diet and bovine blood on reproduction and survivorship in Tabanus nigrovittatus (Diptera: Tabanidae).

    PubMed

    Stoffolano, J G; Majer, J

    1997-11-01

    A synthetic diet, developed for tsetse flies and fed to Tabanus nigrovittatus Macquart before the 2nd gonotrophic cycle, supported complete egg maturation. T. nigrovittatus is autogenous only during the 1st cycle. Overall, 52% of females fed bovine blood and 46% fed the synthetic diet produced mature, stage 10 follicles. Of these, 76% of the blood-fed females and 65% of those fed the synthetic diet laid egg masses, all of which hatched. The median adult survivorship was 9 d for blood-fed and 8 d for those fed the synthetic diet.

  5. Evaluation of Peripheral Blood Smear for Myelodysplasia in Breast Cancer Patients who Received Adjuvant Antracycline.

    PubMed

    Mutlu, Hasan; Akca, Zeki; Teke, Havva Uskudar; Ugur, Hediye

    2011-12-01

    Therapy-related myeloid neoplasms (t-MN) account for approximately 10% to 20% of all cases of AML (acute myeloid leukemia), MDS (myelodysplastic syndrome) and MDS/MPN (myelodysplastic syndrome/myeloproliferative neoplasms), MDS, and MDS/MPN. In our study, we evaluated peripheral blood smear samples and hemogram values in breast cancer patients who were receiving adjuvant anthracycline regimens and were in remission. A total of 78 patients receiving anthracycline-based adjuvant chemotherapy treatment from Kayseri Research and Training Hospital and Mersin State Hospital were enrolled in the study. Their adjuvant treatments had been completed at least 18 months prior to the study. Two patients complained of anemia (2.2%) (Hb<11 mg/dl), leukopenia was observed in seven patients (7.7%) (leukocytes<4000/ mm(3)), and thrombocytopenia was observed in four patients (4.4%) (PLT<150.000/mm(3)). In the blood smear samples, the following were observed: ovalomacrocytes (14%), macrocytes (37%), acanthocytes (1%), stomatocytes (12%), teardrops (12%), nucleated erythrocytes (1%), basophilic stippling (14%), and Howell-Jolly bodies (1%). Additionally, hypo-granulation (38%), Pelger-Huet abnormalities (26%), hypersegmentation (20%), immature granulocytes (8%), and blasts (6%) were observed. We also confirmed the presence of giant platelets (50%) and platelet hypogranulation (19%). According to the peripheral blood smear assessments in our study, we suggest that breast cancer patients should be evaluated for MDS in the early stages, starting from month 18, even if the automated blood counts are normal.

  6. Changes in peripheral blood leukocyte populations in pigs with naturally occurring exudative epidermitis.

    PubMed

    Nofrarías, M; Pujols, J; Segalés, J; Gibert, X; Majó, N

    2006-10-01

    The objective of the study was to analyze changes in peripheral blood leukocyte subsets in cases of naturally occurring exudative epidermitis (EE) in pigs. Five of ten piglets developed the chronic clinical form of EE 2-5 days after weaning (PW). Blood samples were obtained at 7, 14 and 21 days from both normal and clinically affected piglets for routine haematology and for the determination of CD45, CD21, CD4, CD8 and gammadeltaTCR cell markers by flow cytometry. When compared with clinically normal piglets EE affected pigs showed significantly decreased values of monocytes at 14 and 21 days PW, and increased numbers of neutrophils and leukocytes at 21 days PW. The EE affected pigs also had an early significant CD4(+) and CD8(high+) T lymphocyte proliferative response at 7 days PW. However affected pigs had a significantly reduced number of B (CD21(+)) and gammadeltaTCR(+) T lymphocytes in blood at 21 days PW. Although all values remained within the normal range, the significant differences in some peripheral blood leukocyte subsets between the two groups of piglets suggest that the generalised cutaneous infection with Staphylococcus hyicus is severe enough to induce a systemic inflammatory and immune responses.

  7. The oxidative status and inflammatory level of the peripheral blood of rabbits infested with Psoroptes cuniculi

    PubMed Central

    2014-01-01

    Background Psoroptes cuniculi can parasitise the ear canal of the rabbit, and cause the afflicted animals to cease feeding and become severely debilitated, sometimes resulting in death. In this study, we examined the oxidative status and inflammatory level of the peripheral blood of rabbits infested with Psoroptes cuniculi and investigated the pathogenesis of this disease. Methods A total of 24 rabbits were divided into a healthy rabbit group and two infested rabbit groups. After weighing the rabbits, approximately 5 ml of blood was obtained from each animal. Then, the blood serum was extracted and used to assess the levels of antioxidant enzymes and inflammatory factors. Results Compared to the healthy rabbits, the activities of catalase and glutathione-S-transferase and the level of malonyldialdehyde were increased, but the activity of superoxide dismutase was reduced in the infested rabbits. At the same time, a variety of inflammatory cells were activated, and the levels of inflammatory factors such as prostaglandin E2, interleukin-6, interleukin-8 and transforming growth factor-β1 were increased in peripheral blood. Conclusion Animal acariasis was associated with immunosuppressive disorders and inflammatory reaction. These results advance our understanding of the pathogenesis of Psoroptes cuniculi infestation in rabbits and can help guide the effectual treatment of this disease in clinics. PMID:24667000

  8. Novel, high-yield red blood cell production methods from CD34-positive cells derived from human embryonic stem, yolk sac, fetal liver, cord blood, and peripheral blood.

    PubMed

    Olivier, Emmanuel; Qiu, Caihong; Bouhassira, Eric E

    2012-08-01

    The current supply of red blood cells expressing rare blood groups is not sufficient to cover all the existing transfusion needs for chronically transfused patients, such as sickle cell disease homozygous carriers, because of alloimmunization. In vitro production of cultured red blood cells is slowly emerging as a possible complement to the existing collection-based red blood cell procurement system. The yield of cultured red blood cells can theoretically be maximized by amplifying the stem, progenitor, or precursor compartment. Here, we combined methods designed to expand these three compartments to optimize the yield of cultured red blood cells and found that exposing CD34(+) cells to a short pulse of cytokines favorable for erythroid differentiation prior to stem cell expansion followed by progenitor expansion produced the highest yield of erythroid cells. This novel serum-free red blood cell production protocol was efficient on CD34(+) cells derived from human embryonic stem cells, 6-8-week yolk sacs, 16-18-week fetal livers, cord blood, and peripheral blood. The yields of cells obtained with these new protocols were larger by an order of magnitude than the yields observed previously. Globin expression analysis by high-performance liquid chromatography revealed that these expansion protocols generally yielded red blood cells that expressed a globin profile similar to that expected for the developmental age of the CD34(+) cells.

  9. Novel, High-Yield Red Blood Cell Production Methods from CD34-Positive Cells Derived from Human Embryonic Stem, Yolk Sac, Fetal Liver, Cord Blood, and Peripheral Blood

    PubMed Central

    Olivier, Emmanuel; Qiu, Caihong

    2012-01-01

    The current supply of red blood cells expressing rare blood groups is not sufficient to cover all the existing transfusion needs for chronically transfused patients, such as sickle cell disease homozygous carriers, because of alloimmunization. In vitro production of cultured red blood cells is slowly emerging as a possible complement to the existing collection-based red blood cell procurement system. The yield of cultured red blood cells can theoretically be maximized by amplifying the stem, progenitor, or precursor compartment. Here, we combined methods designed to expand these three compartments to optimize the yield of cultured red blood cells and found that exposing CD34+ cells to a short pulse of cytokines favorable for erythroid differentiation prior to stem cell expansion followed by progenitor expansion produced the highest yield of erythroid cells. This novel serum-free red blood cell production protocol was efficient on CD34+ cells derived from human embryonic stem cells, 6–8-week yolk sacs, 16–18-week fetal livers, cord blood, and peripheral blood. The yields of cells obtained with these new protocols were larger by an order of magnitude than the yields observed previously. Globin expression analysis by high-performance liquid chromatography revealed that these expansion protocols generally yielded red blood cells that expressed a globin profile similar to that expected for the developmental age of the CD34+ cells. PMID:23197866

  10. Detecting glycogen in peripheral blood mononuclear cells with periodic acid schiff staining.

    PubMed

    Tabatabaei Shafiei, Mahdieh; Carvajal Gonczi, Catalina M; Rahman, Mohammed Samiur; East, Ashley; François, Jonathan; Darlington, Peter J

    2014-12-23

    Periodic acid Schiff (PAS) staining is an immunohistochemical technique used on muscle biopsies and as a diagnostic tool for blood samples. Polysaccharides such as glycogen, glycoproteins, and glycolipids stain bright magenta making it easy to enumerate positive and negative cells within the tissue. In muscle cells PAS staining is used to determine the glycogen content in different types of muscle cells, while in blood cell samples PAS staining has been explored as a diagnostic tool for a variety of conditions. Blood contains a proportion of white blood cells that belong to the immune system. The notion that cells of the immune system possess glycogen and use it as an energy source has not been widely explored. Here, we describe an adapted version of the PAS staining protocol that can be applied on peripheral blood mononuclear immune cells from human venous blood. Small cells with PAS-positive granules and larger cells with diffuse PAS staining were observed. Treatment of samples with amylase abrogates these patterns confirming the specificity of the stain. An alternate technique based on enzymatic digestion confirmed the presence and amount of glycogen in the samples. This protocol is useful for hematologists or immunologists studying polysaccharide content in blood-derived lymphocytes.

  11. Enumeration of major peripheral blood leukocyte populations for multicenter clinical trials using a whole blood phenotyping assay.

    PubMed

    Hensley, Tiffany R; Easter, Austin B; Gerdts, Sarah E; De Rosa, Stephen C; Heit, Antje; McElrath, M Juliana; Andersen-Nissen, Erica

    2012-09-16

    Cryopreservation of peripheral blood leukocytes is widely used to preserve cells for immune response evaluations in clinical trials and offers many advantages for ease and standardization of immunological assessments, but detrimental effects of this process have been observed on some cell subsets, such as granulocytes, B cells, and dendritic cells. Assaying fresh leukocytes gives a more accurate picture of the in vivo state of the cells, but is often difficult to perform in the context of large clinical trials. Fresh cell assays are dependent upon volunteer commitments and timeframes and, if time-consuming, their application can be impractical due to the working hours required of laboratory personnel. In addition, when trials are conducted at multiple centers, laboratories with the resources and training necessary to perform the assays may not be located in sufficient proximity to clinical sites. To address these issues, we have developed an 11-color antibody staining panel that can be used with Trucount tubes (Becton Dickinson; San Jose, CA) to phenotype and enumerate the major leukocyte populations within the peripheral blood, yielding more robust cell-type specific information than assays such as a complete blood count (CBC) or assays with commercially-available panels designed for Trucount tubes that stain for only a few cell types. The staining procedure is simple, requires only 100 μl of fresh whole blood, and takes approximately 45 minutes, making it feasible for standard blood-processing labs to perform. It is adapted from the BD Trucount tube technical data sheet (version 8/2010). The staining antibody cocktail can be prepared in advance in bulk at a central assay laboratory and shipped to the site processing labs. Stained tubes can be fixed and frozen for shipment to the central assay laboratory for multicolor flow cytometry analysis. The data generated from this staining panel can be used to track changes in leukocyte concentrations over time in

  12. Enumeration of Major Peripheral Blood Leukocyte Populations for Multicenter Clinical Trials Using a Whole Blood Phenotyping Assay

    PubMed Central

    Hensley, Tiffany R.; Easter, Austin B.; Gerdts, Sarah E.; De Rosa, Stephen C.; Heit, Antje; McElrath, M. Juliana; Andersen-Nissen, Erica

    2012-01-01

    Cryopreservation of peripheral blood leukocytes is widely used to preserve cells for immune response evaluations in clinical trials and offers many advantages for ease and standardization of immunological assessments, but detrimental effects of this process have been observed on some cell subsets, such as granulocytes, B cells, and dendritic cells 1-3. Assaying fresh leukocytes gives a more accurate picture of the in vivo state of the cells, but is often difficult to perform in the context of large clinical trials. Fresh cell assays are dependent upon volunteer commitments and timeframes and, if time-consuming, their application can be impractical due to the working hours required of laboratory personnel. In addition, when trials are conducted at multiple centers, laboratories with the resources and training necessary to perform the assays may not be located in sufficient proximity to clinical sites. To address these issues, we have developed an 11-color antibody staining panel that can be used with Trucount tubes (Becton Dickinson; San Jose, CA) to phenotype and enumerate the major leukocyte populations within the peripheral blood, yielding more robust cell-type specific information than assays such as a complete blood count (CBC) or assays with commercially-available panels designed for Trucount tubes that stain for only a few cell types. The staining procedure is simple, requires only 100 μl of fresh whole blood, and takes approximately 45 minutes, making it feasible for standard blood-processing labs to perform. It is adapted from the BD Trucount tube technical data sheet (version 8/2010). The staining antibody cocktail can be prepared in advance in bulk at a central assay laboratory and shipped to the site processing labs. Stained tubes can be fixed and frozen for shipment to the central assay laboratory for multicolor flow cytometry analysis. The data generated from this staining panel can be used to track changes in leukocyte concentrations over time in

  13. Effects of fluoxetine on the oxidative status of peripheral blood leucocytes of restraint-stressed mice.

    PubMed

    Novío, Silvia; Núñez, María Jesús; Amigo, Gonzalo; Freire-Garabal, Manuel

    2011-11-01

    Emotional stress can be viewed as a cause of adverse circumstances that induces a wide range of biochemical and behavioural changes. Oxidative stress is a critical route of damage in various psychological stress-induced disorders such as depression. Antidepressants are widely prescribed to treat these conditions; however, no animal study has investigated the effect of selective serotonin reuptake inhibitors (SSRIs) on the levels of intracellular reactive oxygen species in peripheral blood leucocytes of stressed mice. In this study, mice were immobilized for a period of 6 hr. Fluoxetine (5 mg/kg of body-weight) was administered 30 min. before subjecting the animals to acute stress. The level of intracellular reactive oxygen species in leucocytes of the peripheral blood of stressed mice was investigated using a 2',7'-dichlorofluorescein diacetate probe, and the antioxidant response of fluoxetine was evaluated by superoxide dismutase, diaphorase, catalase and reduced glutathione. Our results show that restraint stress significantly increases the generation of reactive oxygen species in the peripheral defence cells. Treatment with fluoxetine partially reverses the adverse effects of stress. The improvement in cellular oxidative status may be an important mechanism underlying the protective pharmacological effects of fluoxetine, which are clinically observed in the treatment of depressive disorders.

  14. Clinical significance of peripheral blood erythroblastosis after hematopoietic stem cell transplantation.

    PubMed

    Goyama, Susumu; Kanda, Yoshinobu; Nannya, Yasuhito; Ogawa, Seishi; Asano-Moki, Yuki; Ogawa, Natsu; Nakagawa, Masahiro; Sakata-Yanagimoto, Mamiko; Kawazu, Masahito; Komeno, Yukiko; Imai, Yoichi; Hangaishi, Akira; Kurokawa, Mineo; Tsujino, Shiho; Aoki, Katsunori; Chiba, Shigeru; Motokura, Toru; Hirai, Hisamaru

    2004-12-01

    Erythroblasts (EBL) are normally not observed in peripheral blood, but may be found in patients suffering from a variety of severe diseases. The detection of EBL in peripheral blood has been shown to be associated with a poor prognosis. However, the clinical significance of peripheral erythroblastosis after hematopoietic stem cell transplantation (HSCT) has not been evaluated. We retrospectively analyzed the records of 161 patients who underwent HSCT at our hospital from June 1995 to October 2001. EBL at any level were detected in 94% of the patients. Forty-four and 11 patients experienced erythroblastosis exceeding 200 and 1,000/ul, respectively. The erythroblast count was higher in patients who died than in the survivors (geometric mean value 184 vs. 100/ul, P=0.01). High-level erythroblastosis ( >1,000/ul) within 180 days after HSCT was associated with an extremely poor prognosis (median survival 22.5 days). Among the possible confounding factors, the use of total body irradiation (RR 2.35, 95% CI 1.22 - 4.54, P=0.011) and the disease status before transplantation (RR 2.51, 95% CI 1.15 - 5.49, P=0.021) were independent significant factors for erythroblastosis after HSCT. As for post-transplant events, a high EBL concentration was frequently preceded by graft-vs.-host disease, thrombotic microangiopathy, hypoxia, and hematological relapse.

  15. Screening and confirmation of microRNA markers for distinguishing between menstrual and peripheral blood.

    PubMed

    Li, Zhilong; Bai, Peng; Peng, Duo; Wang, Hui; Guo, Yadong; Jiang, Youjing; He, Wang; Tian, Huan; Yang, Yu; Huang, Yuan; Long, Bing; Liang, Weibo; Zhang, Lin

    2017-09-01

    The identification of menstrual blood (MB) and peripheral blood (PB) left at a crime scene is crucial for crime reconstruction, especially in sexual assaults. MicroRNAs (miRNAs), a class of non-protein coding molecules, have been demonstrated to be a viable tool for body fluid identification in forensic casework. Several groups have searched for miRNAs that are specific to different body fluids. Blood has been studied the most extensively. However, menstrual blood was only involved in five studies, and the results confirming the presence of specific miRNAs could not be reproduced in other studies. In this study, we attempted to screen new markers that can differentiate between menstrual blood and peripheral blood by using Exiqon's miRCURY™ LNA Array. Five miRNAs were selected based on the microarray results, namely, miR-141-3p, miR-373-3p, miR-497-5p, miR-143-5p, and miR-136-5p, whose expression levels exhibited 27.95-, 17.96-, 16.74-, 10.14-, and 9.21-fold changes, respectively, compared to the level in peripheral blood. Two classic quantitative methods, TaqMan hydrolysis probes (TaqMan for short) and SYBR Green fluorochrome (SYBR Green for short), were applied in the confirmation step to study the impact of different quantitative methods on the results. Three miRNAs (miR-141-3p, miR-497-5p, and miR-143-5p) were confirmed by TaqMan and one (miR-141-3p) by SYBR Green. Furthermore, bioinformatic methods were applied to interpret the candidate miRNAs. Our results established a multi-step procedure for body fluid identification and showed that the choice of quantitative method is important when miRNAs are used to identify the origin of blood samples. Copyright © 2017 Elsevier B.V. All rights reserved.

  16. DNA methylation array analyses identified breast cancer-associated HYAL2 methylation in peripheral blood.

    PubMed

    Yang, Rongxi; Pfütze, Katrin; Zucknick, Manuela; Sutter, Christian; Wappenschmidt, Barbara; Marme, Frederik; Qu, Bin; Cuk, Katarina; Engel, Christoph; Schott, Sarah; Schneeweiss, Andreas; Brenner, Hermann; Claus, Rainer; Plass, Christoph; Bugert, Peter; Hoth, Markus; Sohn, Christof; Schmutzler, Rita; Bartram, Claus R; Burwinkel, Barbara

    2015-04-15

    Breast cancer (BC) is the leading cause of cancer-related mortality in women worldwide. Changes in DNA methylation in peripheral blood could be associated with malignancy at early stage. However, the BC-associated DNA methylation signatures in peripheral blood were largely unknown. Here, we performed a genome-wide methylation screening and identified a BC-associated differentially methylated CpG site cg27091787 in the hyaluronoglucosaminidase 2 gene (HYAL2) (discovery round with 72 BC case and 24 controls: p = 2.61 × 10(-9) adjusted for cell-type proportions). The substantially decreased methylation of cg27091787 in BC cases was confirmed in two validation rounds (first validation round with 338 BC case and 507 controls: p < 0.0001; second validation round with 189 BC case and 189 controls: p < 0.0001). In addition to cg27091787, the decreased methylation of a 650-bp CpG island shore of HYAL2 was also associated with increased risk of BC. Moreover, the expression and methylation of HYAL2 were inversely correlated with a p-value of 0.006. To note, the BC-associated decreased HYAL2 methylation was replicated in the T-cell fraction (p = 0.034). The cg27091787 methylation level enabled a powerful discrimination of early-stage BC cases (stages 0 and I) from healthy controls [area under curve (AUC) = 0.89], and was robust for the detection of BC in younger women as well (age < 50, AUC = 0.87). Our study reveals a strong association between decreased HYAL2 methylation in peripheral blood and BC, and provides a promising blood-based marker for the detection of early BC. © 2014 UICC.

  17. Postnatal episodic ozone results in persistent attenuation of pulmonary and peripheral blood responses to LPS challenge

    PubMed Central

    Maniar-Hew, Kinjal; Postlethwait, Edward M.; Fanucchi, Michelle V.; Ballinger, Carol A.; Evans, Michael J.; Harkema, Jack R.; Carey, Stephan A.; McDonald, Ruth J.; Bartolucci, Alfred A.

    2011-01-01

    Early life is a dynamic period of growth for the lung and immune system. We hypothesized that ambient ozone exposure during postnatal development can affect the innate immune response to other environmental challenges in a persistent fashion. To test this hypothesis, we exposed infant rhesus macaque monkeys to a regimen of 11 ozone cycles between 30 days and 6 mo of age; each cycle consisted of ozone for 5 days (0.5 parts per million at 8 h/day) followed by 9 days of filtered air. Animals were subsequently housed in filtered air conditions and challenged with a single dose of inhaled LPS at 1 yr of age. After completion of the ozone exposure regimen at 6 mo of age, total peripheral blood leukocyte and polymorphonuclear leukocyte (PMN) numbers were reduced, whereas eosinophil counts increased. In lavage, total cell numbers at 6 mo were not affected by ozone, however, there was a significant reduction in lymphocytes and increased eosinophils. Following an additional 6 mo of filtered air housing, only monocytes were increased in blood and lavage in previously exposed animals. In response to LPS challenge, animals with a prior history of ozone showed an attenuated peripheral blood and lavage PMN response compared with controls. In vitro stimulation of peripheral blood mononuclear cells with LPS resulted in reduced secretion of IL-6 and IL-8 protein in association with prior ozone exposure. Collectively, our findings suggest that ozone exposure during infancy can result in a persistent effect on both pulmonary and systemic innate immune responses later in life. PMID:21131396

  18. MMP-8, MMP-9 and Neutrophil Elastase in Peripheral Blood and Exhaled Breath Condensate in COPD.

    PubMed

    Sng, JieHao Joshua; Prazakova, Silvie; Thomas, Paul S; Herbert, Cristan

    2017-04-01

    Chronic obstructive pulmonary disease (COPD) is characterised by progressive and irreversible airflow limitation associated with chronic inflammation involving cytokines and metalloproteinases (MMPs). MMP-8, MMP-9 and neutrophil elastase (NE) are known to be implicated in COPD but the factors influencing activation and suppression remain unclear. This study aimed to compare MMP-8, MMP-9 and NE in the peripheral blood of COPD patients and controls and to likewise assess exhaled breath condensate (EBC) for these MMPs. Peripheral blood micro(mi)RNA139-5p levels, which may regulate MMPs in COPD, were also measured. Blood and EBC were collected from COPD patients (stable and during exacerbations) and healthy controls. Expression of mRNA for MMP-8, MMP-9, NE and miRNA-139-5p expression in peripheral blood mononuclear cells (PBMCs) was measured using qRT-PCR. MMP-8, MMP-9 and NE protein in plasma as well as MMP-8 and MMP-9 protein in EBC were analysed by enzyme-linked immunoassays. PBMCs from COPD patients showed greater expression of mRNA for MMP-8 (p = 0.0004), MMP-9 (p = 0.0023) and NE (p = 0.0019). PBMC expression of mRNA for NE was significantly higher in COPD exacerbations compared to stable cases (p < 0.05). Expression of mRNA for MMP-9 and NE correlated negatively with spirometry in patients (p < 0.05). Plasma from COPD patients showed greater levels of protein for MMP-8 (p = 0.003), MMP-9 (p = 0.046) and NE (p = 0.018). MMP-8 protein levels were lower in the EBC of COPD patients (p < 0.0001). In PBMCs, enhanced expression of mRNA for MMP-9 and NE is associated with COPD and may correlate with disease severity and exacerbations.

  19. Disseminated histiocytic sarcoma with peripheral blood involvement in a Bernese Mountain dog.

    PubMed

    Rossi, Silvia; Gelain, Maria Elena; Comazzi, Stefano

    2009-03-01

    A 6-year-old Bernese Mountain dog was presented with a history of lethargy and weight loss of 2 weeks duration. On physical examination the dog had pale mucous membranes and tachypnea. Ultrasound examination revealed hepatomegaly, splenomegaly, and mesenteric lymphadenomegaly. Results of a CBC included marked normocytic normochromic nonregenerative anemia, marked thrombocytopenia, and moderate leukocytosis with mild neutrophilia and a large population of unclassified round cells (6.2 x 10(3)/microL). The unclassified cells occasionally were bi- or multinucleated and had variably abundant pale basophilic cytoplasm that contained multiple irregular clear vacuoles and occasionally erythrocytes. Fine needle aspirate specimens of the mesenteric lymph nodes and spleen were composed of a population of round pleomorphic cells with the same features as the circulating cells. On flow cytometric analysis of peripheral blood, the unclassified cells expressed CD18, CD45, CD11c, CD1c, and CD14; immunocytochemical analysis of blood smears also indicated the cells were positive for CD1c, CD1a, and CD11c. The dog died a few hours after referral. The histologic interpretation of samples collected from spleen, liver, and lymph nodes was malignant neoplasia of histiocytic origin. Immunohistochemical staining yielded negative results for CD11d, a marker of red-pulp macrophages, ruling out hemophagocytic histiocytic sarcoma. Based on clinical and pathologic findings, the final diagnosis was disseminated histiocytic sarcoma (DHS) with peripheral blood involvement. To our knowledge, DHS in a dog with evidence and immunophenotyping of neoplastic cells in peripheral blood has been reported only rarely.

  20. Effect of Contaminating Red Blood Cells on OKT3-Mediated Polyclonal Activation of Peripheral Blood Mononuclear Cells

    PubMed Central

    Song, Samuel; Goodwin, Joseph; Zhang, Jenny; Babbitt, Bruce; Lathey, Janet L.

    2002-01-01

    Erythrocytes are typically present as impurities in the majority of peripheral blood mononuclear cell (PBMC) preparations. This study was undertaken to investigate the effects of contaminating red blood cells (RBC) on the ability of OKT3 to activate CD4+ and CD8+ T cells. Surprisingly, the levels of gamma interferon, tumor necrosis factor alpha, and interleukin-1β (IL-1β) produced by PBMC upon stimulation by OKT3 were increased (P < 0.05) in a dose-dependent manner when increasing amounts of autologous RBC (RBC-to-PBMC ratios of 2:1, 10:1, and 50:1) were spiked into PBMC preparations. The OKT3-driven induction of the IL-2 receptor (CD25) and the proliferation of T lymphocytes in response to phorbol myristate acetate were not affected by the addition of RBC. PMID:11986282

  1. Acute modulation of cytokine gene expression in bovine peripheral blood mononuclear cells (PBMCs) by endogenous cortisol

    USDA-ARS?s Scientific Manuscript database

    Cortisol suppresses many aspects of immune function. However, recent publications suggest acute cortisol exposure may actually enhance immune function (Dhabhar. 2009. Neuroimmunomod. 16:300). The objective of this study was to determine the influence of acute increases in endogenous cortisol on expr...

  2. Effects of Methylprednisolone Sodium Succinate on Clearance of Live E. coli from the Peripheral Blood of Dogs.

    DTIC Science & Technology

    1978-08-28

    live E . coli organisms from peripheral blood of dogs. The experimental group was pretreated with 30 mg/kg of MP while controls received equal volumes...of saline. Both control and MP pretreated dogs significantly reduced the number of E . coli in peripheral blood by almost two orders of magnitude...however, there was no significant difference in clearance of E . coli organisms between the two groups. An initial leukopenia occured in both groups after

  3. [Research advances on DNA extraction methods from peripheral blood mononuclear cells].

    PubMed

    Wang, Xiao-Ying; Yu, Chen-Xi

    2014-10-01

    DNA extraction is a basic technology of molecular biology. The purity and the integrality of DNA structure are necessary for different experiments of gene engineering. As commonly used materials in the clinical detection, the fast, efficient isolation and extraction of genomic DNA from peripheral blood mononuclear cells is very important for the inspection and analysis of clinical blood. At present, there are many methods for extracting DNA, such as phenol-chloroform method, salting out method, centrifugal adsorption column chromatography method (artificial methods), magnetic beads (semi-automatic method) and DNA extraction kit. In this article, a brief review of the principle for existing DNA blood extraction method, the specific steps and the assessment of the specific methods briefly are summarized.

  4. Flow cytometric analysis of dengue virus-infected cells in peripheral blood.

    PubMed

    Baclig, Michael O; Gervacio, Leonora T S; Suarez, Lady-Anne C; Buerano, Corazon C; Matias, Ronald R; Kumatori, Atsushi; Inoue, Shingo; Morita, Kouichi; Natividad, Filipinas F; Hasebe, Futoshi

    2010-11-01

    With the development of permeabilization techniques in flow cytometry and the availability of various monoclonal antibodies (MAbs) that specifically bind with cell surface and intracellular antigens, it is now possible to use flow cytometric assay to identify dengue virus (DEN) infected cells in peripheral blood. Blood samples were analyzed using phycoerythrin (PE) labeled anti-CD3, anti-CD14, anti-CD16, and anti-CD19 antibodies and Alexa Fluor 488 labeled anti-flavivirus monoclonal antibody (MAb) 6B6C-1. The predominant DEN-infected cells were CD19+ in this study. There was dim partial to moderately bright partial expression of CD19 positive cells in the blood samples tested. Virus isolation and serotype-specific RT-PCR revealed the cells were infected with dengue serotype 3 (DEN3). Our results suggest B cells may play an important role in DEN1 and DEN3 replication, and dissemination in vivo.

  5. Detection of Leishmania amastigotes in peripheral blood from four dogs--Short communication.

    PubMed

    Giudice, Elisabetta; Passantino, Annamaria

    2011-06-01

    The authors carried out microscopic examination of blood smears of 1438 dogs infected with Leishmania infantum. Unusual findings of leishmaniosis associated with circulating parasitised cells are described in four dogs. Most of the dogs presented severe illness, with lethargy, dysorexia, emaciation and alterations of the haematological pattern (anaemia, thrombocytopenia, neutrophilia and monocytosis). In three cases, leishmaniosis was associated with ehrlichiosis. On examination of peripheral blood smears, Leishmania sp. amastigotes were observed both in various circulating leukocytes (neutrophil, monocyte, macrophage) and free. In conclusion, parasites can rarely be detected in blood smears (in 0.28% of the animals examined); thus, the time-consuming microscopic search for amastigotes can make only a weak contribution to the conventional diagnosis of canine leishmaniosis.

  6. Dihydroergotoxine decreases blood pressure in spontaneously hypertensive rats by interacting with peripheral dopamine receptors.

    PubMed

    Memo, M; Sagheddu, G; Carruba, M O; Spano, P

    1985-04-22

    Dihydroergotoxine (10 micrograms/kg s.c.) decreased mean carotid blood pressure in urethane-anaesthetized spontaneously hypertensive rats but failed to modify the same parameter in normotensive rats. The effect was statistically significant 20 min after the injection and relatively long lasting (up to 90 min). Pharmacological characterization of the phenomenon indicated that it is mediated by stimulation of dopamine receptors, since pretreatment with haloperidol, cis-flupentixol but not with trans-flupentixol, completely prevent the reduction in blood pressure induced by dihydroergotoxine. Moreover, a challenge dose of dihydroergotoxine did not reduce mean blood pressure values in spontaneously hypertensive rats pretreated with domperidone or (-)sulpiride, but not with (+)sulpiride. These results suggest that the ergot derivative modifies the cardiovascular system by interaction with peripheral dopamine receptors of the DA2 type.

  7. A new surface marker on T lymphocytes of human peripheral blood.

    PubMed

    Hammarström, S; Hellström, U; Perlmann, P; Dillner, M L

    1973-11-01

    Neuraminidase treatment of human peripheral blood lymphocytes uncovers cell surface receptors that bind purified A hemagglutinin from the snail Helix pomatia. No hemagglutinin was bound to untreated lymphocytes. Binding studies with (125)I-labeled hemagglutinin suggested that the number of receptors on neuraminidase-treated lymphocytes was approximately 1.10(6)/cell. The apparent association constant for hemagglutinin binding to lymphocytes, as calculated from Scatchard's plots, was 5-7.10(8) liters/mol. Immunofluorescent staining with FITC-conjugated hemagglutinin gave positive reactions with approximately 60% of the lymphocytes from normal donors. Positive staining was inversely related to the number of lymphocytes with Fc or complement receptors or with surface immunoglobulin, thus suggesting that See PDF for Structure the lymphocytes with receptors for Helix pomatia A hemagglutinin are T cells. Cell fractionation on columns charged with hemagglutinin indicate that these receptors may be used for separating subpopulations of human peripheral lymphocytes.

  8. Integrated Bioinformatics Approach Reveals Crosstalk Between Tumor Stroma and Peripheral Blood Mononuclear Cells in Breast Cancer.

    PubMed

    He, Lang; Wang, Dan; Wei, Na; Guo, Zheng

    2016-01-01

    Breast cancer is now the leading cause of cancer death in women worldwide. Cancer progression is driven not only by cancer cell intrinsic alterations and interactions with tumor microenvironment, but also by systemic effects. Integration of multiple profiling data may provide insights into the underlying molecular mechanisms of complex systemic processes. We performed a bioinformatic analysis of two public available microarray datasets for breast tumor stroma and peripheral blood mononuclear cells, featuring integrated transcriptomics data, protein-protein interactions (PPIs) and protein subcellular localization, to identify genes and biological pathways that contribute to dialogue between tumor stroma and the peripheral circulation. Genes of the integrin family as well as CXCR4 proved to be hub nodes of the crosstalk network and may play an important role in response to stroma-derived chemoattractants. This study pointed to potential for development of therapeutic strategies that target systemic signals travelling through the circulation and interdict tumor cell recruitment.

  9. T cell receptor repertoire in the peripheral blood and intestinal mucosa of coeliac patients.

    PubMed Central

    Lahat, N; Ben-Nun, A; Cohen, L; Kinarty, A; Lerner, A

    1995-01-01

    The alpha beta and gamma delta T cell receptor (TCR) repertoire in the peripheral blood and intestinal mucosa of six coeliac and six age-matched controls was analysed by reverse transcription and polymerase chain reaction (PCR). No TCR alpha and gamma delta restriction was observed in coeliacs and controls. However, V gamma 3 was expressed only in coeliac peripheral and intestinal T cells. V delta 2 was strongly expressed in coeliacs and scarcely transcribed in control cells. The unique expression of these gamma delta TCR in coeliac patients suggests that V gamma 3 and perhaps V delta 2 TCR-bearing lymphocytes may play a role in the pathogenesis of coeliac disease. Images Fig. 1 Fig. 2 Fig. 3 Fig. 4 Fig. 5 Fig. 6 PMID:7664488

  10. [The role of peripheral blood eosinophil percentage in classification of chronic rhinosinusitis with nasal polyps].

    PubMed

    Wang, Ming-jie; Zhou, Bing; Li, Yun-chuan; Huang, Qian

    2013-08-01

    Trying to find a useful marker to subclassify chronic rhinosinusitis (CRS), ratio of eosinophil in peripheral blood was investigated. Histologic characteristics of surgical samples were analyzed in 119 CRS with nasal polyps (CRSwNP) patients, who were classified into eosinophil CRSwNP (ECRSwNP) group and non-ECRSwNP group. Peripheral blood eosinophil percentage, olfactory function, skin prick test, serum total IgE and sinus CT scan in two groups were all examined and analyzed. To evaluate the discriminatory power of eosinophil to diagnose ECRSwNP, the Receiver Operating Characteristic(ROC)curve was analysed. There were significant differences in Ratio of EOS, serum total IgE, and olfactory function scores, between ECRSwNP group and non-ECRSwNP group(mean value were 7.31%: 3.90%, 60.9 IU/L: 28.9 IU/L, 5.8: 0.4 respectively, U value were 620.01, 1020.53 and 1092.52, respectively, all P < 0.05). However, there was no difference in skin prick test between two groups. In CT scan exam, there were no differences in Lund-Mackay scores in frontal sinus, anterior ethmoid sinus, posterior ethmoid sinus, sphenoid sinus and ostiomeatal complex area, but maxillary sinus, between ECRSwNP group and non-ECRSwNP group (U value were 27.5, 23.5, 22.5, 31.5, 28.5, respectively, all P > 0.05, and U value of maxillary sinus was 12.01, P < 0.05 ). Peripheral blood eosinophil percentage and serum total IgE were related with pathology of nasal polyps (r value were 0.55, 0.24, and P value were 0.001, 0.01, respectively), especially blood eosinophilia can be a predictor of ECRSwNP. The area under curve was 0.818 and cutoff value was 5.65%. ECRSwNP is different from non-ECRSwNP in many clinical features. Peripheral blood eosinophil percentage is consistent with histologic features of ECRS, which is a useful marker as 5.65% in classification of CRS.

  11. Wall morphology, blood flow and wall shear stress: MR findings in patients with peripheral artery disease.

    PubMed

    Galizia, Mauricio S; Barker, Alex; Liao, Yihua; Collins, Jeremy; Carr, James; McDermott, Mary M; Markl, Michael

    2014-04-01

    To investigate the influence of atherosclerotic plaques on femoral haemodynamics assessed by two-dimensional (2D) phase-contrast (PC) magnetic resonance imaging (MRI) with three-directional velocity encoding. During 1 year, patients with peripheral artery disease and an ankle brachial index <1.00 were enrolled. After institutional review board approval and written informed consent, 44 patients (age, 70 ± 12 years) underwent common femoral artery MRI. Patients with contra-indications for MRI were excluded. Sequences included 2D time-of-flight, proton-density, T1-weighted and T2-weighted MRI. Electrocardiogram (ECG)-gated 2D PC-MRI with 3D velocity encoding was acquired. A radiologist classified images in five categories. Blood flow, velocity and wall shear stress (WSS) along the vessel circumference were quantified from the PC-MRI data. The acquired images were of good quality for interpretation. There were no image quality problems related to poor ECG-gating or slice positioning. Velocities, oscillatory shear stress and total flow were similar between patients with normal arteries and wall thickening/plaque. Patients with plaques demonstrated regionally increased peak systolic WSS and enhanced WSS eccentricity. Combined multi-contrast morphological imaging of the peripheral arterial wall with PC-MRI with three-directional velocity encoding is a feasible technique. Further study is needed to determine whether flow is an appropriate marker for altered endothelial cell function, vascular remodelling and plaque progression. • Femoral plaques are associated with altered dynamics of peripheral blood flow. • Multi-contrast MRI can investigate the presence and type of atherosclerotic plaques. • Three-dimensional velocity-encoding phase-contrast MRI can investigate flow and wall shear stress. • Atherosclerotic peripheral arteries demonstrate increased systolic velocities and wall shear stress.

  12. Dopamine D5 receptor expression is unchanged in peripheral blood lymphocytes in essential hypertension.

    PubMed

    Ricci, A; Chiandussi, L; Schena, M; Schiavone, D; Veglio, F; Amenta, F

    1995-11-01

    The present study was designed to investigate possible changes in the expression of lymphocyte dopamine receptor in essential hypertension. The expression of dopamine D5 receptor was evaluated by radioligand binding techniques using [3H]-SCH 23390 as ligand. Plasma catecholamines, aldosterone levels and plasma renin activity were also measured. Eleven borderline hypertensive patients, 15 patient with the mild essential hypertension, 7 patients with moderate essential hypertension and 5 patients with severe essential hypertension were examined. Plasma catecholamine levels were assayed by high pressure liquid chromatography with electrochemical detection. Dopamine D5 receptor was measured by radioligand binding techniques. Plasma aldosterone levels and renin activity were determined by radio immunoassay. [3H]-SCH 23390 was specifically bound to human peripheral blood lymphocytes. The binding was time-, temperature- and concentration-dependent with a dissociation constant (Kd) value of 0.59 nM and a maximum density of binding sites (Bmax) of 223 pmol/10(6) cells. Dopamine competed with [3H]-SCH 23390 binding in the submicromolar range suggesting the labelling of a dopamine D5 receptor. No changes in the density of [3H]-SCH 23390 binding sites were observed in human peripheral blood lymphocytes between essential hypertensive patients and normotensive subjects. Also catecholamines, plasma renin activity and aldosterone levels were unchanged. In spite of the availability of a sensitive technique for measuring dopamine receptors in human peripheral lymphocytes, no change in their expression was noticeable in essential hypertension. This suggests that dopamine receptor analysis in essential hypertension is not a useful marker for investigating hypertension-dependent changes of the peripheral dopaminergic system.

  13. Controlled ovarian hyperstimulation induced changes in the expression of circulatory miRNA in bovine follicular fluid and blood plasma.

    PubMed

    Noferesti, Sina Seifi; Sohel, Md Mahmodul Hasan; Hoelker, Michael; Salilew-Wondim, Dessie; Tholen, Ernst; Looft, Christian; Rings, Franca; Neuhoff, Christiane; Schellander, Karl; Tesfaye, Dawit

    2015-12-09

    Despite its role in increasing the number of offspring during the lifetime of an individual animal, controlled ovarian hyperstimulation (COH) may have detrimental effects on oocyte development, embryo quality and endometrial receptivity. Circulating miRNAs in bio-fluids have been shown to be associated with various pathological conditions including cancers. Here we aimed to investigate the effect of COH on the level of extracellular miRNAs in bovine follicular fluid and blood plasma and elucidate their mode of circulation and potential molecular mechanisms to be affected in the reproductive tract. Twelve simmental heifers were estrous synchronized and six of them were hyperstimulated using FSH. Follicular fluid samples from experimental animals were collected using ovum pick up technique at day 0 of the estrous cycle and blood samples were collected at day 0, 3 and 7 of post ovulation. The expression profile of circulatory miRNAs in follicular fluid and blood plasma were performed using the human miRCURY LNA™ Universal RT miRNA PCR array system. A comparative threshold cycle method was used to determine the relative abundance of the miRNAs. A total of 504 and 402 miRNAs were detected in both bovine follicular fluid and blood plasma, respectively. Of these 57 and 21 miRNAs were found to be differentially expressed in follicular fluid and blood plasma, respectively derived from hyperstimulated versus unstimulated heifers. Bioinformatics analysis of those circulating miRNAs indicated that their potential target genes are involved in several pathways including TGF-beta signaling pathway, MAPK signaling pathway, pathways in cancer and Oocyte meiosis. Moreover, detail analysis of the mode of circulation of some candidates showed that most of the miRNA were found to be detected in both exosomal and Ago2 protein complex fraction of both follicular fluid and blood plasma. Our data provide the consequence of hyperstimulation induced changes of extracellular miRNAs in

  14. Blood Contamination of the Small Bore Peripheral Intravenous Catheter in Neonates.

    PubMed

    Krishnamurthy, Vani; Doreswamy, Srinivasa Murthy; Krishnagowda, Sushma

    2016-11-01

    Peripheral Intravenous Catheters (PIV) are extensively used in sick neonates for administration of medicines and nutrition. When these PIVs are used on intermittent basis, they are flushed with saline in order to keep the hub of the catheter free from blood. Presence of blood in the hub of the catheter can be potentially dangerous as it could facilitate infection. The aim of this study was to find the magnitude of blood contamination of PIV catheter hub after routine flushing. We measured the volume of 24 g PIV by filling it with saline and thereby measuring its volume. The PIVs which were in situ for at least 6 hours and removed were used for this study. These catheters were flushed with 0.2 ml of saline and the RBC count was calculated. A total of 94 PIVs were studied, out of which 84% showed blood tinged residual flush and 15% of them had visible blood clot. All (100%) of the catheter studied showed RBCs on microscopic examination. The median RBC count was 36960/cu mm and the interquartile range was 10000 - 113920/cu mm. The highest RBC count was 2080000/cu mm. Blood contamination of the small bore PIVs after flushing is universal in neonates.

  15. Blood Contamination of the Small Bore Peripheral Intravenous Catheter in Neonates

    PubMed Central

    Doreswamy, Srinivasa Murthy; Krishnagowda, Sushma

    2016-01-01

    Introduction Peripheral Intravenous Catheters (PIV) are extensively used in sick neonates for administration of medicines and nutrition. When these PIVs are used on intermittent basis, they are flushed with saline in order to keep the hub of the catheter free from blood. Presence of blood in the hub of the catheter can be potentially dangerous as it could facilitate infection. Aim The aim of this study was to find the magnitude of blood contamination of PIV catheter hub after routine flushing. Materials and Methods We measured the volume of 24 g PIV by filling it with saline and thereby measuring its volume. The PIVs which were in situ for at least 6 hours and removed were used for this study. These catheters were flushed with 0.2 ml of saline and the RBC count was calculated. Results A total of 94 PIVs were studied, out of which 84% showed blood tinged residual flush and 15% of them had visible blood clot. All (100%) of the catheter studied showed RBCs on microscopic examination. The median RBC count was 36960/cu mm and the interquartile range was 10000 – 113920/cu mm. The highest RBC count was 2080000/cu mm. Conclusion Blood contamination of the small bore PIVs after flushing is universal in neonates. PMID:28050372

  16. High Fibrinogen in Peripheral Blood Correlates with Poorer Hearing Recovery in Idiopathic Sudden Sensorineural Hearing Loss

    PubMed Central

    Kanzaki, Sho; Sakagami, Masafumi; Hosoi, Hiroshi; Murakami, Shingo; Ogawa, Kaoru

    2014-01-01

    Objectives We used hearing tests and peripheral blood sample analyses to characterize the pathology of idiopathic sudden sensorineural hearing loss (ISSNHL) and to identify possible prognostic factors for predicting recovery of hearing loss. Study Design A retrospective, multicenter trial was conducted. Methods Two hundred three patients examined within 7 days after the onset of ISSNHL received prednisone with lipo-prostaglandin E1. Pure-tone auditory tests were performed before and after treatment with these drugs. Blood tests were performed on blood samples collected during the patients’ initial visit to our clinic. Results In all patients, elevated white blood cell (WBC) counts, fasting blood sugar levels, HgbA1c, and erythrocyte sedimentation rate (ESR) significantly correlated with high hearing threshold measurements obtained on the initial visit. High fibrinogen levels, WBC counts, ESR, and low concentrations of fibrinogen degradation products (FDP) were associated with lower hearing recovery rates. Additionally, different audiogram shapes correlated with different blood test factors, indicating that different pathologies were involved. Conclusions High fibrinogen levels measured within seven days after ISSNHL onset correlated with poorer hearing recovery. This may be a consequence of ischemia or infections in the inner ear. The high WBC counts also observed may therefore reflect an immune response to inner ear damage induced by ischemic changes or infections. Our data indicate that therapeutic strategies should be selected based on the timing of initial treatment relative to ISSNHL onset. PMID:25166620

  17. Peripheral blood mammaglobin gene expression for diagnosis and prediction of metastasis in breast cancer patients.

    PubMed

    Radwan, Wafaa M; Moussa, Heba S; Essa, Enas S; Kandil, Samia H; Kamel, Azza M

    2013-03-01

    To evaluate the value of peripheral blood mammaglobin (MG) gene expression for diagnosis and prediction of metastasis in breast cancer patients. MG expression was detected by nested reverse-transcription polymerase chain reaction in the peripheral blood of 46 females (32 breast cancer, 12 benign breast lesions, 2 no breast abnormalities). In total 28 breast cancer patients were followed up through a period of 34 months for the development of metastasis. MG expression was detected in 16/32 (50%) breast cancer patients but not in patients with benign lesions or healthy participants. Five patients had metastasis at diagnosis. During the 34 months of follow up, five more MG-positive patients showed metastatic lesions and none of the MG negative patients who were followed up developed metastasis. The study suggests blood MG expression is a specific molecular marker for detection of occult mammary carcinoma cells of patients with operable breast cancer. It might be of value as a predictor of subsequent metastasis. Large-scale studies and longer follow-up periods are needed. © 2012 Wiley Publishing Asia Pty Ltd.

  18. Peripheral white blood cells profile of biodegradable metal implant in mice animal model

    NASA Astrophysics Data System (ADS)

    Paramitha, Devi; Noviana, Deni; Estuningsih, Sri; Ulum, Mokhamad Fakhrul; Nasution, Ahmad Kafrawi; Hermawan, Hendra

    2015-09-01

    Biocompatibility or safety of the medical device is considered important. It can be determined by blood profile examination. The aim of this study was to assess the biocompatibility of biodegradable metal implant through peripheral white blood cells (WBCs) profile approach. Forty eight male ddy mice were divided into four groups according to the materials implanted: iron wire (Fe), magnesium rod (Mg), stainless steel surgical wire (SS316L) and control with sham (K). Implants were inserted and attached onto the right femoral bone on latero-medial region. In this study, peripheral white blood cells and leukocyte differentiation were the parameters examined. The result showed that the WBCs value of all groups were decreased at the first day after implantation, increased at the 10th day and continued increasing at the 30th day of observation, except Mg group which has decreased. Neutrophil, as an inflammatory cells, was increased at the early weeks and decreased at the day-30 after surgery in all groups. Despite, these values during the observation were still within the normal range. As a conclus ion, biodegradable metal implants lead to an inflammatory reaction, with no adverse effect on WBC value found.

  19. Inhibition of peripheral blood neutrophil oxidative burst in periodontitis patients with a homeopathic medication Traumeel S.

    PubMed

    Žilinskas, Juozas; Žekonis, Jonas; Žekonis, Gediminas; Šadzevičienė, Renata; Sapragonienė, Marija; Navickaitė, Justina; Barzdžiukaitė, Ingrida

    2011-05-01

    The anti-inflammatory effects of a homeopathic remedy, Traumeel S, have been observed in experimental and clinical studies; however, its antioxidant properties have not been elucidated. The aim of the present study was to evaluate the antioxidant effects of Traumeel S on peripheral blood neutrophils in patients with periodontitis. The study was performed using venous blood of 22 individuals with chronic periodontitis and 21 healthy subjects. The antioxidant effects of Traumeel S on the production of reactive oxygen species by unstimulated and stimulated with unopsonized E. coli neutrophils were investigated using luminol- and lucigenin-dependent chemiluminescence (CL). Polymorphonuclear leukocytes of periodontitis patients produced higher levels (p<0.01) of light output of lucigenin-dependent chemiluminescence and significantly reduced (p<0.01) light output of luminol-dependent chemiluminescence than analogous cells of healthy subjects. Highly diluted (10⁻⁴ of the stem solution) Traumeel S significantly (by approximately 50%) reduced superoxide-induced oxidation of lucigenin by unstimulated and stimulated with unopsonized E. coli polymorphonuclear leukocytes of periodontitis patients and had a tendency to intensify luminol-dependent chemiluminescence. Preincubation of the unstimulated and stimulated with unopsonized E. coli polymorphonuclear leukocytes of healthy subjects with Traumeel S exerts no inhibitory action on the luminol- and lucigenin-dependent chemiluminescence of the above-mentioned cells. This study indicates that Traumeel S may significantly reduce production of superoxide anion by unstimulated and stimulated peripheral blood polymorphonuclear neutrophils of periodontitis patients.

  20. Femoral catheters: safety and efficacy in peripheral blood stem cell collection.

    PubMed

    Adorno, G; Zinno, F; Bruno, A; Lanti, A; Ballatore, G; Masi, M; Cudillo, L; Del Poeta, G; Riccitelli, A; Del Principe, M I; Pepe, R; Marchitelli, E; Morosetti, M; Meloni, C; Isacchi, G; Amadori, S

    1999-10-01

    Central venous access is necessary in patients candidate for peripheral blood stem cell (PBSC) collection. We report our experience with a dual lumen femoral catheter (Gamcath, 11 french), initially designed for hemodialysis. We studied 147 patients and performed 488 collections after mobilization with either G-CSF alone or chemotherapy + G-CSF, when the white blood cell count exceeded 1 x 10(9)/L, or when a measurable population of CD34+ cells (20/microL) was detected in peripheral blood. All patients received systemic anticoagulation with a low weight heparin and ultrasound examination was performed after the removal of the catheter. Seven patients developed thrombosis (4.7%), ten experienced hematomas at the site of catheter placement (6.8%) despite prophylactic platelet transfusions, while only one patient (0.6%) had a catheter-related infection. In conclusion, the short-term use of large bore femoral catheters in setting up PBSC collection seems to be associated with minimal risk of infection and low thrombotic incidence.

  1. Development of implantable autologous small-calibre vascular grafts from peripheral blood samples.

    PubMed

    Aper, T; Teebken, O E; Krüger, A; Heisterkamp, A; Hilfiker, A; Haverich, A

    2013-04-01

    At present the generation of a small-calibre (≤5 mm) vascular replacement for artificial bypasses remains a challenge for tissue engineering. The biocompatibility of bioartificial vessel replacements is of decisive significance for function and depends on the materials used. A completely autologous vessel substitute must exhibit high biocompatibility and functionality. For this purpose we developed and optimised a technique for the engineering of an autologous bypass material from a fibrin scaffold and vascular cells isolated from the same sample of peripheral blood in a porcine model. Fibrinogen, late outgrowth endothelial and smooth muscle cells were isolated from peripheral blood samples (n=14, 100 mL each). Fibroblasts were isolated from porcine aortic adventitial tissue (n=4). Tubular seeded fibrin segments were obtained using an injection moulding technique with the simultaneous incorporation of the in vitro expanded cells into the fibrin matrix. The segments were cultivated under dynamic conditions with pulsatile perfusion in a bioreactor. Morphological and functional characterization was done. Artificial vascular segments with a length of 150 mm were reproducibly obtained with a hierarchical arrangement of incorporated cells similar to the structure of the vascular wall. By additional seeding of fibroblasts, suturable segments with biomechanical properties suitable for implantation into the arterial system were obtained. Implantable bioartificial vascular grafts can be generated from blood. After cultivation under dynamic conditions the vascular segments possess a structure similar to that of the vascular wall and exhibit biomechanical properties sufficient for implantation as arterial substitutes. Georg Thieme Verlag KG Stuttgart · New York.

  2. Basis of monitoring central blood pressure and hemodynamic parameters by peripheral arterial pulse waveform analyses.

    PubMed

    Miyashita, Hiroshi; Katsuda, Shin-ichiro

    2013-01-01

    In hypertension clinics, central blood pressure (CBP) should be estimated, instead of directly measured, by the "signal processing" of a noninvasive peripheral pressure waveform. This paper deals with the data obtained in our three separate studies focusing on a major estimation method, i.e., radial artery late systolic shoulder pressure (rSBP2)-based CBP estimation. Study 1: Using a wave separation analysis of precise animal data of pressure wave transmission along the upper-limb arteries, we first demonstrate that pulse pressure amplification is largely attributable to local wave reflection alone. Study 2: A frequency component analysis of simultaneously recorded human central and radial artery pressure waveforms showed a predominance of lower (1st+2nd) harmonic components in determining the central augmentation peak amplitude. The features of a central pressure waveform, including its phase property, may contribute to the less-altered transmission of augmentation peak pressure to rSBP2. Study 3: Comparisons of noninvasive rSBP2 with direct or estimated central systolic blood pressure (cSBP) revealed broad agreement but also augmentation-dependent biases. Based on the features of the biases as well as the counterbalanced relationship between pulse pressure amplification and the transmission-induced alterations of augmentation peak amplitude observed in Study 2, we propose an improved cSBP estimate, SBPm, the simple arithmetic mean of rSBP2 and peripheral systolic blood pressure.

  3. Characterization of porcine peripheral blood leukocytes by light-scattering flow cytometry.

    PubMed Central

    Wang, F I; Williams, T J; el-Awar, F Y; Pang, V F; Hahn, E C

    1987-01-01

    As a basis for other experiments using flow cytometry of porcine peripheral blood leukocytes, cell fractions were isolated by various methods and analyzed by forward angle light scatter and 90 degree light scatter. Cytospin smears of cell samples were also studied by leukocyte differential counts and nonspecific esterase staining. Three main populations of peripheral blood leukocytes [lymphocytes, monocytes, and granulocytes (primarily neutrophils)], were defined in the log 90 degree light scatter by forward angle light scatter histogram. Partial overlap was observed between lymphocyte and monocyte, and between monocyte and granulocyte domains. Correlation between leukocyte differential counts and flow cytometric quantification based on bitmap statistics of appropriate domains was between r = 0.872-0.892 for lymphocyte and granulocyte. Percoll density gradients were used for subfractionation of leukocyte populations, especially for the enrichment of granulocytes. The specific densities were calculated for lymphocytes (1.0585-1.0819 g/cc), monocytes (1.0585-1.0702 g/cc), granulocyte (1.0819-1.0936 g/cc), and erythrocytes (greater than 1.0952 g/cc). We suggest that light scatter characterization is a basis for future studies of porcine blood by flow cytometry. PMID:3453262

  4. Determination of hematopoietic stem cells in peripheral blood by an automated hematology analyzer (SE-9000).

    PubMed

    Takekawa, K; Yamane, T; Hino, M; Tatsumi, N

    1998-12-01

    We evaluated the usefulness of an automated hematology analyzer (SE-9000) for the identification and counting of peripheral blood stem cells (PBSCs). The samples tested were from 14 patients with hematological malignancies. Peripheral blood samples were collected from the subjects before and after a course of chemotherapy. From the leukapheresis sample, CD34+ cells, assumed to be hematopoietic stem cells, were obtained with an immunomagnetic cell separator. The CD34+ cells obtained accumulated in the gate corresponding to low recurrent frequencies of the automated hematology analyzer. This gate shows results of the 'immature information' (IMI) channel. Software for detection of only the cells that accumulated in this gate was therefore developed. With this trial program, the regression coefficient between the percentage of leukocytes from the blood samples that were CD34+ and the percentage of such leukocytes that appeared on the IMI channel was 0.79. With this analyzer, the number of PBSC could be counted in about 80 s. The identification and counting of cells picked up by the IMI channel should be clinically useful for the monitoring of changes in PBSC after chemotherapy for mobilization.

  5. [Peripheral blood cells luminol-dependent chemiluminescence at the different stages of atopic dermatitis].

    PubMed

    Elistratova, I V; Morozov, S G; Zakharova, I A; Tarasova, M V

    2015-01-01

    Aim of this work was to record the luminol-dependent spontaneous and induced chemiluminescence at the different stages of atopic dermatitis. Peripheral blood cells were obtained from adult patient with atopic dermatitis followed by the registration of luminol-dependent chemiluminescence on luminograph. Opsonized zymosan as well as yeasts Candida tropicalis have been used to induce the chemiluminescence. Spontaneous and induced chemiluminescence were slightly elevated at the mild atopic dermatitis but were decreased at the severe stage of disease. Statistically significant difference has been found between group with mild and severe atopic dermatitis, Skin contamination by yeasts Candida tropicalis causes the increased level of blood cells chemiluminescence at the first week of atopic relapse when the disease was mild. Severe stage of atopic dermatitis was coupled with statistically significant inhibition of both, spontaneous and induced chemiluminescence. Luminol-dependent chemiluminescence of peripheral blood cells from adult atopic dermatitis patients may be stimulated at the mild stage and suppressed at severe stage of atopic dermatitis.

  6. Inhibition of peripheral blood neutrophil oxidative burst in periodontitis patients with a homeopathic medication Traumeel S

    PubMed Central

    žilinskas, Juozas; žekonis, Jonas; žekonis, Gediminas; Šadzevičienė, Renata; Sapragonienė, Marija; Navickaitė, Justina; Barzdžiukaitė, Ingrida

    2011-01-01

    Summary Background The anti-inflammatory effects of a homeopathic remedy, Traumeel S, have been observed in experimental and clinical studies; however, its antioxidant properties have not been elucidated. The aim of the present study was to evaluate the antioxidant effects of Traumeel S on peripheral blood neutrophils in patients with periodontitis. Material/Methods The study was performed using venous blood of 22 individuals with chronic periodontitis and 21 healthy subjects. The antioxidant effects of Traumeel S on the production of reactive oxygen species by unstimulated and stimulated with unopsonized E. coli neutrophils were investigated using luminol- and lucigenin-dependent chemiluminescence (CL). Results Polymorphonuclear leukocytes of periodontitis patients produced higher levels (p<0.01) of light output of lucigenin-dependent chemiluminescence and significantly reduced (p<0.01) light output of luminol-dependent chemiluminescence than analogous cells of healthy subjects. Highly diluted (10−4 of the stem solution) Traumeel S significantly (by approximately 50%) reduced superoxide-induced oxidation of lucigenin by unstimulated and stimulated with unopsonized E. coli polymorphonuclear leukocytes of periodontitis patients and had a tendency to intensify luminol-dependent chemiluminescence. Preincubation of the unstimulated and stimulated with unopsonized E. coli polymorphonuclear leukocytes of healthy subjects with Traumeel S exerts no inhibitory action on the luminol- and lucigenin-dependent chemiluminescence of the above-mentioned cells. Conclusions This study indicates that Traumeel S may significantly reduce production of superoxide anion by unstimulated and stimulated peripheral blood polymorphonuclear neutrophils of periodontitis patients. PMID:21525811

  7. The effect of melatonin on peripheral blood cells during total body irradiation in rats.

    PubMed

    Koc, Mehmet; Buyukokuroglu, Mehmet Emin; Taysi, Seyithan

    2002-05-01

    Melatonin, has been reported to participate in the regulation of a number of important physiological and pathological process. It has also the ability to protect the genetic material of hematopoietic cells of mice from damaging effects of acute total body irradiation. The objective of this study was to the potential radioprotective effects of pharmacological doses of melatonin in total body irradiated rat's peripheral blood cells. Forty adult rats were divided into 4 equal groups. Group 1 received no melatonin or irradiation (control group), while group 2 received only melatonin (5 mg/kg, i.p.). Group 3 received only total body irradiation (RT) by 5 Gy of gamma irradiation only and group 4 received RT plus melatonin (5 mg/kg, i.p., 30 min before RT). An hour and a half following RT, blood samples were taken. Leukocytes and thrombocytes number and hemoglobin levels were measured in all groups. Five mg/kg dose of melatonin significantly protected leukocytes and as well as thrombocytes number against y irradiation. There were no significant differences between Hb levels. Our results suggest that melatonin administration prior to irradiation prevented radiation damage on peripheral blood cells. Melatonin radioprotection is achieved by its ability as a scavenger for free radicals generated by ionizing radiation and acts probably as a growth factor, especially for granulocytes in bone marrow.

  8. Peripheral blood eosinophilia in association with generalized pustular and erythrodermic psoriasis.

    PubMed

    Mansur, A T; Göktay, F; Yaşar, S P

    2008-04-01

    In recent studies, it has been documented that the eosinophil cells play active role in many kinds of inflammatory disorders. Measurements of the mediators released by eosinophils and cell counts in serum and skin have provided some evidence indicating the role of eosinophils in psoriasis. To evaluate the blood eosinophil cell count in patients with erythrodermic psoriasis and generalized pustular psoriasis. In this study, 48 patients with histopathologically proven psoriasis (33 with erythrodermic, 15 with generalized pustular); 43 patients with maculopapular, erythematous, or bullous drug eruption with widespread involvement; and 51 patients with basal cell carcinoma were included. These three groups were compared with each other in terms of their absolute eosinophil cell counts and percentage of eosinophils. Forty-one point seven per cent of patients with psoriasis had peripheral blood eosinophilia compared with 58.1% of the patients with drug eruption and 11.8% of patients with basal cell carcinoma. The percentage of patients with eosinophilia both in patients suffering from psoriasis and drug eruption were significantly increased compared with the patients with basal cell carcinoma (P < 0.001). The absolute median eosinophil cell counts both in patients with psoriasis and drug eruption were also significantly differed from patients with basal cell carcinoma (259.2, 439.1 and 183.1, respectively; P < 0.001) Peripheral blood eosinophilia seems to be associated with severe forms of psoriasis. This finding may suggest that the eosinophils have significant roles in the pathogenesis of these types of psoriasis.

  9. Serotonin Uptake Is Largely Mediated by Platelets versus Lymphocytes in Peripheral Blood Cells

    PubMed Central

    2012-01-01

    The serotonin transporter (SERT), a primary target for many antidepressants, is expressed in the brain and also in peripheral blood cells. Although platelet SERT function is well accepted, lymphocyte SERT function has not been definitively characterized. Due to their small size, platelets often are found in peripheral blood mononuclear cell preparations aimed at isolating lymphocytes, monocytes, and macrophages. The presence of different cells makes it difficult to assign SERT expression and function to specific cell types. Here, we use flow cytometry and IDT307, a monoamine transporter substrate that fluoresces after uptake into cells, to investigate SERT function in lymphocyte and platelet populations independently, as well as simultaneously without prior isolation. We find that murine lymphocytes exhibit temperature-dependent IDT307 transport but uptake is independent of SERT. Lack of measurable SERT function in lymphocytes was corroborated by chronoamperometry using serotonin as a substrate. When we examined rhesus and human mixed blood cell populations, we found that platelets, and not lymphocytes, were primary contributors to SERT function. Overall, these findings indicate that lymphocyte SERT function is minimal. Moreover, flow cytometry, in conjunction with the fluorescent transporter substrate IDT307, can be widely applied to investigate SERT in platelets from populations of clinical significance. PMID:23336055

  10. Comparative cell morphology in the peripheral blood film from exotic and native animals.

    PubMed

    Canfield, P J

    1998-12-01

    Haematological investigation is an important part of disease diagnosis. This is particularly so when investigating individual animal disease. It may also be important when investigating diseases in groups of animals, but the opportunity to perform necropsies to directly detect diseases processes often diminishes its usefulness. Haematological investigation is essentially similar for all species. The presence of nucleated erythrocytes and thrombocytes in nonmammals requires alteration of haemoglobin measurement and cell counting. In addition, it may cause some confusion in identification of cells in peripheral blood films. Examination of blood films is an important component of haematological investigation and provides useful information on erythroid, leukocytic and platelet/thrombocytic alterations. Interpretation of alterations is essentially similar for all species. However, cell identification can at times be difficult. There are five basic leukocytes in all species: neutrophil (mammals) or heterophil (nonmammals), eosinophil, basophil, lymphocyte and monocyte. In nonmammals it may be difficult at times to distinguish between heterophils and eosinophils. In addition, lymphocytes may be confused with thrombocytes. However, a common-sense approach to the examination of the peripheral blood film will minimise this confusion.

  11. Differential gene expression associated with inflammation in peripheral blood cells of patients with pneumoconiosis

    PubMed Central

    Liu, Shu-jun; Wang, Ping; Jiao, Jie; Han, Lin; Lu, Yu-min

    2016-01-01

    Objectives: To study expression changes in inflammation-related genes in peripheral blood of patients with pneumoconiosis and to explore the possibility of these genes as pneumoconiosis biomarkers. Methods: Peripheral blood samples of patients with pneumoconiosis patients and controls were collected, and total RNA of the blood cells were extracted and reverse transcribed to cDNA. Screenings of deferentially expressed genes associated with inflammation between patients with pneumoconiosis and controls were performed using real-time quantitative PCR array and the expressions of the three most upregulated genes were confirmed by real-time PCR. Results: The expression of 11 genes was significantly altered in patients with pneumoconiosis compared with those of the control. Among these 11 genes, 8 genes were upregulated and 3 were downregulated. Preliminary results indicated that interleukin 6 (IL-6) mRNA expression in patients with pneumoconiosis was higher than that in controls (P=0.019). The level of IL6 mRNA expression in the patients was higher than that in non-smoking controls, but it was neither affected by type and stage of pneumoconiosis nor by time of contact with dust. Conclusions: IL6 was possibly involved in the development of pneumoconiosis. PMID:27265534

  12. Peripheral white blood cells profile of biodegradable metal implant in mice animal model

    SciTech Connect

    Paramitha, Devi; Noviana, Deni Estuningsih, Sri; Ulum, Mokhamad Fakhrul; Nasution, Ahmad Kafrawi; Hermawan, Hendra

    2015-09-30

    Biocompatibility or safety of the medical device is considered important. It can be determined by blood profile examination. The aim of this study was to assess the biocompatibility of biodegradable metal implant through peripheral white blood cells (WBCs) profile approach. Forty eight male ddy mice were divided into four groups according to the materials implanted: iron wire (Fe), magnesium rod (Mg), stainless steel surgical wire (SS316L) and control with sham (K). Implants were inserted and attached onto the right femoral bone on latero-medial region. In this study, peripheral white blood cells and leukocyte differentiation were the parameters examined. The result showed that the WBCs value of all groups were decreased at the first day after implantation, increased at the 10th day and continued increasing at the 30th day of observation, except Mg group which has decreased. Neutrophil, as an inflammatory cells, was increased at the early weeks and decreased at the day-30 after surgery in all groups. Despite, these values during the observation were still within the normal range. As a conclus ion, biodegradable metal implants lead to an inflammatory reaction, with no adverse effect on WBC value found.

  13. Clinical application and evaluation of Sysmex XE-5000 analyser for detecting nucleated red blood cells in peripheral blood.

    PubMed

    Zheng, Shan-Luan; Zheng, Tian; Cheng, Xiang; Hao, Xiao-Ke

    2014-01-01

    To evaluate the performance of Sysmex XE-5000 analyser for detecting nucleated red blood cells (NRBCs) in peripheral blood and investigate the clinical application of this analyser. The absolute NRBC counts (NRBC#) and percentage (NRBC%) of 137 blood specimens (NRBC-positive according to the DIFF channel of the analyser) were determined in the NRBC channel of the analyser. The intra-assay imprecision, carryover rate, and linear range of the analyser were evaluated. The NRBC% of the blood sample was detected with a microscope, and the difference between two methods was analysed. The intra-assay imprecision of the analyser for detecting NRBC# in specimens with high, moderate, and low Q-flag values were 2.10%, 3.26%, and 11.62%, respectively, and the imprecision for detecting NRBC% were 3.79%, 5.80%, and 13.33%, respectively. The carryover rates of the analyser for detecting NRBC# and NRBC% were 0.51% and 0.26%, respectively. Sysmex XE-5000 analyser had good linearity in NRBC# (i.e., 0/L to 18 x 10(9)/L). The NRBC%s of the two methods did not significantly differ (p = 0.716). The analyser can completely replace the traditional microscope for clinically classifying and counting NRBCs.

  14. Successful in vitro antigen-dependent activation of 24-hour-old peripheral blood lymphocytes.

    PubMed

    Owen, J A; Muirhead, K; Jensen, C; Jonak, Z L

    1996-03-28

    We describe a simple, rapid and reproducible in vitro culture system in which human peripheral blood lymphocytes (PBLs), donated 24 h prior to initiation of culture can be stimulated to produce antigen-specific antibodies. Peripheral blood lymphocytes purified by Ficoll-Hypaque centrifugation were passed over a G10 Sephadex column and then activated in vitro in the presence of 0.003% staphylococcus Cowan A, 2.8 x 10(-6) M indomethacin and appropriate concentrations of tetanus toxoid antigen. After the first 24 h in culture, a five-fold concentrated supernatant from an allogeneic mixed lymphocyte culture was added. The cell surface phenotypes of the PBLs were analyzed by flow cytometry at the initiation and termination of culture, in order to provide a comprehensive characterization of the cellular composition of a successful in vitro stimulation system. Our results clearly show that the majority of peripheral blood B cells can be induced to an activated stage (blast transformation) and interleukin 2 (IL-2) receptor expression, following very simple manipulations of the lymphoid population. Tetanus toxoid-specific antibody production can be readily generated in this cell population. In contrast, T cells were not activated to express IL-2 receptors and reach blast transformation, and did not show appreciable proliferation. Our system provides a population of B cells producing antibodies of desired specificity which could be utilized for the generation of human hybridomas or could serve as a donor population for antibody engineering via the combinatorial library approach. Careful light scattering and cell surface phenotypic analyses of the cells entering, proliferating and differentiating in these cultures enabled several novel observations to be made.

  15. Gene Expression Differences in Peripheral Blood of Parkinson’s Disease Patients with Distinct Progression Profiles

    PubMed Central

    Soreq, Lilach; Lobo, Patrícia P.; Mestre, Tiago; Coelho, Miguel; Rosa, Mário M.; Gonçalves, Nilza; Wales, Pauline; Mendes, Tiago; Gerhardt, Ellen; Fahlbusch, Christiane; Bonifati, Vincenzo; Bonin, Michael; Miltenberger-Miltényi, Gabriel; Borovecki, Fran; Soreq, Hermona; Ferreira, Joaquim J.; F. Outeiro, Tiago

    2016-01-01

    The prognosis of neurodegenerative disorders is clinically challenging due to the inexistence of established biomarkers for predicting disease progression. Here, we performed an exploratory cross-sectional, case-control study aimed at determining whether gene expression differences in peripheral blood may be used as a signature of Parkinson’s disease (PD) progression, thereby shedding light into potential molecular mechanisms underlying disease development. We compared transcriptional profiles in the blood from 34 PD patients who developed postural instability within ten years with those of 33 patients who did not develop postural instability within this time frame. Our study identified >200 differentially expressed genes between the two groups. The expression of several of the genes identified was previously found deregulated in animal models of PD and in PD patients. Relevant genes were selected for validation by real-time PCR in a subset of patients. The genes validated were linked to nucleic acid metabolism, mitochondria, immune response and intracellular-transport. Interestingly, we also found deregulation of these genes in a dopaminergic cell model of PD, a simple paradigm that can now be used to further dissect the role of these molecular players on dopaminergic cell loss. Altogether, our study provides preliminary evidence that expression changes in specific groups of genes and pathways, detected in peripheral blood samples, may be correlated with differential PD progression. Our exploratory study suggests that peripheral gene expression profiling may prove valuable for assisting in prediction of PD prognosis, and identifies novel culprits possibly involved in dopaminergic cell death. Given the exploratory nature of our study, further investigations using independent, well-characterized cohorts will be essential in order to validate our candidates as predictors of PD prognosis and to definitively confirm the value of gene expression analysis in aiding

  16. Seasonal influence on mitogen and cyclosporin responses of peripheral blood lymphocytes.

    PubMed

    Michelis, Fotios V; Delitheos, Andreas K; Tiligada, Ekaterini

    2013-06-01

    The immune response and lymphocyte activation in particular are affected by environmental factors. In vivo and in vitro experiments demonstrate variability in lymphocyte activation according to seasonal changes. This study focused on the effects of season on the ex vivo mitogen-induced activation of lymphocytes from peripheral blood of healthy humans living in a temperate climate, as well as the ex vivo lymphocyte activation of rabbits living under constant laboratory conditions. The possible impact of season on the action of the immunosuppressant drug cyclosporin A (CsA) on lymphocyte activation was investigated in both species. Cultured peripheral blood lymphocytes from human donors (n=13, 22-63years of age) and from animals housed under 12:12hour light:dark cycle were stimulated with phytohemagglutinin (PHA) in the absence or presence of 10 and 25μg/mL CsA. Lymphocyte activation was assessed by morphometric analysis under a light microscope. Percentages of unactivated lymphocytes, activated lymphoblasts and aberrant cells reflecting cytotoxicity were determined. Human lymphocytes demonstrated a significant decrease in response to PHA during the winter months, in comparison to the rest of the year. In contrast, the peripheral blood lymphocytes of rabbits housed under constant conditions did not demonstrate similar variations in response to PHA stimulation. The immunosuppressive action of cyclosporin A on this experimental model was unaffected by the observed seasonal variation in mitogen response in humans. These findings may guide research towards the identification of factors associated with the seasonality of the immune response and its potential influence on therapeutic interventions.

  17. The peripheral blood of Aβ binding RBC as a biomarker for diagnosis of Alzheimer's disease.

    PubMed

    Lan, Jie; Liu, Jinping; Zhao, Zhuo; Xue, Rong; Zhang, Nan; Zhang, Peilan; Zhao, Peng; Zheng, Fang; Sun, Xuguo

    2015-05-01

    in vitro, it has been reported that amyloid β (Aβ) is bound to red blood cells (RBCs) and this process damages the red cell. Also, a possible relationship between RBCs and Alzheimer's disease (AD) is supported by the findings of RBC impairment in AD. Therefore, Aβ fibrils bounding RBC are of great interest as potential biomarkers. in this study, we focused on Aβ amyloid fibrils and/or aggregation on the peripheral RBC from 50 subjects with AD and 50 healthy controls (HCs) through thioflavin T (ThT) staining followed by immunofluorescence assay to confirm the presence of Aβ amyloid fibrils and/or aggregation on the RBC. Then we optimised fluorescence staining and imaging conditions and analysed the images obtained by image processing software. we have analysed RBC morphology in blood from 50 subjects with AD and 50 HCs found that 16.8% of the RBCs are elongated as compared with 6.7% in normal controls (P < 0.01), and there is a negative correlation between the two parameters (P < 0.05). Our study showed that 98% of AD peripheral RBCs were amyloid binding-positive (ranging from 2 to 30%), while only 38% that of RBCs (ranging from 2 to 3.4%) were in HCs. We also found four modified morphologies of RBCs triggered by Aβ binding, which may serve as an ancillary investigation and indicate the progression of AD. we first directly prove the existence of Aβ binding RBCs in peripheral blood. In addition, we observed new modified morphologies of RBC triggered by Aβ binding, all of those can serve as a biomarker for the diagnosis and progression of AD. © The Author 2015. Published by Oxford University Press on behalf of the British Geriatrics Society. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

  18. Brain amyloidosis ascertainment from cognitive, imaging, and peripheral blood protein measures

    PubMed Central

    Hwang, Kristy S.; Avila, David; Elashoff, David; Kohannim, Omid; Teng, Edmond; Sokolow, Sophie; Jack, Clifford R.; Jagust, William J.; Shaw, Leslie; Trojanowski, John Q.; Weiner, Michael W.; Thompson, Paul M.

    2015-01-01

    Background: The goal of this study was to identify a clinical biomarker signature of brain amyloidosis in the Alzheimer's Disease Neuroimaging Initiative 1 (ADNI1) mild cognitive impairment (MCI) cohort. Methods: We developed a multimodal biomarker classifier for predicting brain amyloidosis using cognitive, imaging, and peripheral blood protein ADNI1 MCI data. We used CSF β-amyloid 1–42 (Aβ42) ≤192 pg/mL as proxy measure for Pittsburgh compound B (PiB)-PET standard uptake value ratio ≥1.5. We trained our classifier in the subcohort with CSF Aβ42 but no PiB-PET data and tested its performance in the subcohort with PiB-PET but no CSF Aβ42 data. We also examined the utility of our biomarker signature for predicting disease progression from MCI to Alzheimer dementia. Results: The CSF training classifier selected Mini-Mental State Examination, Trails B, Auditory Verbal Learning Test delayed recall, education, APOE genotype, interleukin 6 receptor, clusterin, and ApoE protein, and achieved leave-one-out accuracy of 85% (area under the curve [AUC] = 0.8). The PiB testing classifier achieved an AUC of 0.72, and when classifier self-tuning was allowed, AUC = 0.74. The 36-month disease-progression classifier achieved AUC = 0.75 and accuracy = 71%. Conclusions: Automated classifiers based on cognitive and peripheral blood protein variables can identify the presence of brain amyloidosis with a modest level of accuracy. Such methods could have implications for clinical trial design and enrollment in the near future. Classification of evidence: This study provides Class II evidence that a classification algorithm based on cognitive, imaging, and peripheral blood protein measures identifies patients with brain amyloid on PiB-PET with moderate accuracy (sensitivity 68%, specificity 78%). PMID:25609767

  19. [Enrichment and Biological Characteristics of Peripheral Blood-derived Mesenchymal Stem Cells in Rats].

    PubMed

    Jia, Lin-Ying; Zhang, Qian; Fang, Ning; Chen, Long; Yu, Li-Mei; Liu, Jin-Wei; Zhang, Tao

    2015-04-01

    To explore the effective method for enrichment of rat peripheral blood-derived mesenchymal stem cells(PBMSC) and study the cell biological characteristics. Peripheral mononuclear cells were isolated by density gradient centrifugation from blood of 4 week old rats after G-CSF mobilization. Thereafter, the fibroblast-like cells were acquired by plastic-adherent culture, and the proliferation curve was assayed. For analyzing surface markers of the second generation cultured isolated PBMSC, both flow cytometry(CD90, CD44, CD29, CD45, CD11b and CD79a) and immunocytochemical staining(CD73, CD105, CD34 and HLA-DR) methods were used. Furthermore, the differentiation capacities of PBMSC into osteocytes, chondrocytes and adipocytes were identified. (1) The adherent cells displayed typical colony-forming unit fibroblast(CFU-F) growth pattern after 6-7 day of primary culture and reached 80% confluence after 21 days of culture. The passaged PBMSC possessed high proliferative capacity and spindle growth pattern and was able to grown into exponential phase next day with a doubling time of 39.2 h. (2) PBMSC expressed mesenchymal markers such as CD90, CD44, CD29, CD73 and CD105, but failed to expressed markers of CD45, CD11b, CD79a, CD34 and HLA-DR. (3) After 21 days of culture in osteogenic, chondrogenic and adipogenic differentiation media, calcifying nodules, intracellular glycosaminoglycans and lipid droplets could be found by alizarin red, alcian blue and oil red-O staining, respectively. PBMSC can be enriched from rat peripheral blood with high purity and abundance by our methods. The growth and phenotypic characteristics of the isolated PBMSC are consistent with that of well-known MSC, and these cells possess the capability to multi-lineage mesoderm differentiation.

  20. Antibody forming cells and plasmablasts in peripheral blood in CVID patients after vaccination.

    PubMed

    Chovancova, Zita; Vlkova, Marcela; Litzman, Jiri; Lokaj, Jindrich; Thon, Vojtech

    2011-05-31

    Common variable immunodeficiency (CVID), the most frequent primary antibody disorder, is characterized by hypogammaglobulinaemia and impaired antibody production. Poor vaccination response is essential for the diagnosis of CVID. Their under laying defects remain to be elucidated. Routine determination of antibody production in serum from CVID patients after vaccination and investigation of B cell function in vivo is complicated due to substitution therapy. Therefore we investigated antibody production on the B-cell level by ELISPOT and characterized changes in B-cell subpopulations in CVID patients, including plasmablasts, in peripheral blood by flow cytometry after vaccination for specification of the diagnosis. Thirty-seven CVID patients and eighty healthy volunteers were immunized with tetanus toxoid and pneumococcal polysaccharide vaccines. Specific antibody levels and B cell subpopulations were measured before vaccination and on day 7 after vaccination by ELISPOT assay and flow cytometry respectively. Of the thirty-seven well defined CVID patients studied, thirty lacked detectable spot forming cells producing specific IgG, IgA or IgM antibodies against employed vaccines and seven had only weak responses compared to controls. In the control group, an increase in circulating plasmablasts on day 7 post immunization corresponded with the appearance of antibody forming cells. In contrast, CVID patients failed to increase plasmablasts significantly in peripheral blood after antigen challenge. Our findings indicate that CVID patients have a block in terminal B-cell differentiation and that flow based assessment of plasmablasts in peripheral blood after vaccination serves as a surrogate diagnostic marker for assessing in vivo antibody responses in patients suspected to have CVID. Copyright © 2011 Elsevier Ltd. All rights reserved.

  1. Tc1/Tc2 imbalance in the peripheral blood of patients with recurrent genital herpes.

    PubMed

    Deng, Yunhua; Yang, Dongliang; Chen, Xingping; Chen, Yinling

    2006-01-01

    In order to investigate the IFN-gamma and IL-4 expression of CD8+ T lymphocytes in the peripheral blood from patients with recurrent genital herpes (RGH) at different clinical periods and their relationship with the pathogenesis of RGH, flow cytometry was used to detect the intracellular cytokines (IFN-gamma and IL-4) of CD8+ T lymphocytes in the peripheral blood of 30 patients with RGH at acute period, 20 patients with RGH at recovery period and 15 healthy volunteers. The results showed that RGH patients at acute period had a lower percentage of Tc1 subsets in peripheral blood than that of healthy controls (P < 0.001), especially a remarkable decreased percentage of Tc1 subsets (P < 0.001) among those RGH patients with recurrent number more than 3 in the recent half a year. Tc1/Tc2 ratio in the RGH patients at acute period was significantly decreased as compared with normal control group (P < 0.05). The recurrent number of acute patients in the recent half a year was significantly correlated with the percentage of Tc1 subsets and the ratio of Tc1/Tc2 (P < 0.05). A decreased percentage of Tc1 subsets was found among the RGH patients with recurrent number more than 3 in the recent half a year at recovery period in comparison with healthy volunteers (P < 0.05), and it was significantly correlated with the recurrent number in the recent half a year (P < 0.05). It is concluded that there are Tc1/Tc2 imbalance and a low level of Tc1 subsets in RGH patients who are relapsing repeatedly in the near period. The low level of Tc1 subsets may be an important factor for the recurrence of RGH and the reactivation of latent herpesvirus infection.

  2. Brain amyloidosis ascertainment from cognitive, imaging, and peripheral blood protein measures.

    PubMed

    Apostolova, Liana G; Hwang, Kristy S; Avila, David; Elashoff, David; Kohannim, Omid; Teng, Edmond; Sokolow, Sophie; Jack, Clifford R; Jagust, William J; Shaw, Leslie; Trojanowski, John Q; Weiner, Michael W; Thompson, Paul M

    2015-02-17

    The goal of this study was to identify a clinical biomarker signature of brain amyloidosis in the Alzheimer's Disease Neuroimaging Initiative 1 (ADNI1) mild cognitive impairment (MCI) cohort. We developed a multimodal biomarker classifier for predicting brain amyloidosis using cognitive, imaging, and peripheral blood protein ADNI1 MCI data. We used CSF β-amyloid 1-42 (Aβ42) ≤ 192 pg/mL as proxy measure for Pittsburgh compound B (PiB)-PET standard uptake value ratio ≥ 1.5. We trained our classifier in the subcohort with CSF Aβ42 but no PiB-PET data and tested its performance in the subcohort with PiB-PET but no CSF Aβ42 data. We also examined the utility of our biomarker signature for predicting disease progression from MCI to Alzheimer dementia. The CSF training classifier selected Mini-Mental State Examination, Trails B, Auditory Verbal Learning Test delayed recall, education, APOE genotype, interleukin 6 receptor, clusterin, and ApoE protein, and achieved leave-one-out accuracy of 85% (area under the curve [AUC] = 0.8). The PiB testing classifier achieved an AUC of 0.72, and when classifier self-tuning was allowed, AUC = 0.74. The 36-month disease-progression classifier achieved AUC = 0.75 and accuracy = 71%. Automated classifiers based on cognitive and peripheral blood protein variables can identify the presence of brain amyloidosis with a modest level of accuracy. Such methods could have implications for clinical trial design and enrollment in the near future. This study provides Class II evidence that a classification algorithm based on cognitive, imaging, and peripheral blood protein measures identifies patients with brain amyloid on PiB-PET with moderate accuracy (sensitivity 68%, specificity 78%). © 2015 American Academy of Neurology.

  3. Effect of graphene oxide on bacteria and peripheral blood mononuclear cells.

    PubMed

    Campos-Delgado, Jessica; Castro, Kelly L S; Munguia-Lopez, Jose G; González, Ana K; Mendoza, Martin E; Fragneaud, Benjamin; Verdan, Raphael; Araujo, Joyce R; González, Francisco J; Navarro-Contreras, Hugo; Pérez-Maldonado, Ivan N; León-Rodríguez, Antonio de; Achete, Carlos A

    2016-11-02

    Driven by the potential biological applications of graphene, many groups have studied the response of cells exposed to graphene oxide (GO). In particular, investigations of bacteria indicate that there are 2 crucial parameters, which so far have only been investigated separately: GO size and exposure methodology. Our study took into account both parameters. We carefully characterized the samples to catalog sizes and structural properties, and tested different exposure methodologies: exposure in saline solution and in the presence of growth media. Furthermore, we performed experiments with peripheral blood mononuclear cells exposed to our GO materials. Atomic force microscopy, scanning electron microscopy, Raman spectroscopy, X-ray photoelectron spectroscopy and transmission electron microscopy were used to characterize the morphology and composition of different samples of GO: GO-H2O, GO-PBS and GO-MG. Our samples had 2D sizes of ~100 nm (GO-H2O and GO-PBS) and >2 µm (GO-MG). We tested antibacterial activity and cytotoxicity toward peripheral blood mononuclear cells of 3 different GO samples. A size-dependent growth inhibition of Escherichia coli (DH5 α) in suspension was found, which proved that this effect depends strongly on the protocol followed for exposure. Hemocompatibility was confirmed by exposing peripheral blood mononuclear cells to materials for 24 hours; viability and apoptosis tests were also carried out. Our experiments provide vital information for future applications of GO in suspension. If its antibacterial properties are to be potentiated, care should be taken to select 2D sizes in the micrometer range, and exposure should not be carried out in the presence of grow media.

  4. [Exacerbations in perimenstrual asthma. Clinical significance of peripheral blood eosinophilia and BMI].

    PubMed

    Skoczyński, Szymon; Semik-Orzech, Aleksandra; Pierzchała, Ewa; Rzepka-Wrona, Patrycja; Kołodziejczyk, Krzysztof; Pierzchała, Władysław

    2016-01-01

    Asthma is a heterogeneous disease characterized by lower airways' obstruction, caused by various factors. There are many asthma phenotypes. Lately, perimenstrual asthma (PMA) with a pattern of exacerbations before and during menstruation as well as obesity associated asthma have been a subject of particular scientific and clinical interest. 30 women were qualified for this three-arm case-control study(women with a pattern of asthma exacerbations in the perimenstrual period, women with asthma but no perimenstrual exacerbations, healthy control group). All patients performed spirometry and assessed disease control using specific questionnaires. Peripheral blood counts with smear were also performed. PMA patients differ in a statistically significant way in respect of anthropometric measurements such as BMI: in PMA group 25.8±1.8; in non-PMA asthmatics 23.9 ±2.2; healthy control 23.1±1.5; p=0.018) and spirometry results (FEV1 [%]: 85.1 (36.3-113.0); in PMA asthmatics, 93.1 (81,6-109,7), in nonPMA group, p<0.05; 105.4 (108,3-119,0) in healthy control and Tiffeneau index [%]: 70.1 (41.2-98.1); in PMA vs 83.5 (59.6-94.4); in non-PMA asthmatics 93.1(81,8-97,5) in healthy control p<0.05; ). PMA asthmatics also complain of poorer disease control than non-PMA asthmatics. There were no differences in peripheral blood eosinophilia or CRP between studied groups, p>0.05). Asthma exacerbations are not associated with the effect of peripheral blood eosinophilia. Women with greater BMI are more predisposed to perimenstrual asthma.

  5. Increased levels of LPS-binding protein in bovine blood and milk following bacterial lipopolysaccharide challenge.

    PubMed

    Bannerman, Douglas D; Paape, Max J; Hare, William R; Sohn, Eun Jung

    2003-10-01

    Several species of gram-negative bacteria, including Escherichia coli, Klebsiella pneumoniae, and various species of Enterobacter, are common mastitis pathogens. All of these bacteria are characterized by the presence of endotoxin or lipopolysaccharide (LPS) in their outer membrane. The bovine mammary gland is highly sensitive to LPS, and LPS has been implicated, in part, in the pathogenesis of gram-negative mastitis. Recognition of LPS is a key event in the innate immune response to gram-negative infection and is mediated by the accessory molecules CD14 and LPS-binding protein (LBP). The objective of the current study was to determine whether LBP levels increased in the blood and mammary gland following LPS challenge. The left and right quarters of five midlactating Holstein cows were challenged with either saline or LPS (100 microg), respectively, and milk and blood samples collected. Basal levels of plasma and milk LBP were 38 and 6 microg/ml, respectively. Plasma LBP levels increased as early as 8 h post-LPS challenge and reached maximal levels of 138 microg/ ml by 24 h. Analysis of whey samples derived from LPS-treated quarters revealed an increase in milk LBP by 12 h. Similar to plasma, maximal levels of milk LBP (34 microg/ml) were detected 24 h following the initial LPS challenge. Increments in milk LBP levels paralleled a rise in soluble CD14 (sCD14) levels and initial rises in the levels of these proteins were temporally coincident with maximal neutrophil recruitment to the inflamed gland. Because LBP and sCD14 are known to enhance LPS-induced host cell activation and to facilitate detoxification of LPS, these data are consistent with a role for these molecules in mediating mammary gland responses to LPS.

  6. Copper and zinc concentrations in the uterine fluid and blood serum during the bovine estrous cycle.

    PubMed

    Alavi-Shoushtari, Sayed Mortaza; Asri Rezaie, Siamak; Pak, Mozhgan; Alizadeh, Sajad; Abedizadeh, Roya; Khaki, Amir

    2012-01-01

    To investigate uterine and serum copper (Cu) and zinc (Zn) concentrations variation during the bovine estrus cycle , 232 blood and genital tract samples were collected from the abattoir in Urmia. The phase of the estrous cycle was determined by the examination of the ovaries and the uterine tonicity. Of the 46 samples selected for use in the study, 13 were pro-estrus, 10 estrus, 8 metestrus, and 15 diestrus. The uterus was incised and uterine fluid was collected by gentle scraping of the uterine mucosa with a curette. The total mean (± SEM) Cu concentrations in serum and uterine fluid samples, determined by spectrophotometry, were 66.1± 6.5 and 171.3 ± 33.2µg dL(-1) respectively, which were significantly different, while total mean serum and uterine fluid Zn concentrations were 91.9 ± 5.4 and 291.6 ± 23.4 µg dL(-1), which also showed a significant difference. The mean serum Cu values in different phases of the estrous cycle were not significantly different, while uterine fluid Cu content in pro-estrus and diestrus were significantly higher than those in estrus and metestrus, and were also significantly higher than those of the serum samples. The mean Zn value of serum samples at different stages of the cycle was not significantly different. The mean Zn value of the uterine fluid samples was also not significantly different in different stages, but in pro-estrus, metestrus, and in diestrus they were highly significantly different from those of the serum. These results showed that Cu concentrations in the uterine fluid vary at different stages of the cycle and are higher than those in the blood serum, but, the uterine Zn content does not vary during the estrous cycle and is much higher than those in the serum, that seems to be due to the secretory action of the uterine mucosa.

  7. Peripheral blood DNA methylation profiles are indicative of head and neck squamous cell carcinoma

    PubMed Central

    Langevin, Scott M.; Koestler, Devin C; Christensen, Brock C; Butler, Rondi A; Wiencke, John K; Nelson, Heather H; Houseman, E Andres; Marsit, Carmen J

    2012-01-01

    Head and neck cancer accounts for an estimated 47,560 new cases and 11,480 deaths annually in the United States, the majority of which are squamous cell carcinomas (HNSCC). The overall 5 year survival is approximately 60% and declines with increasing stage at diagnosis, indicating a need for non-invasive tests that facilitate the detection of early disease. DNA methylation is a stable epigenetic modification that is amenable to measurement and readily available in peripheral blood. We used a semi-supervised recursively partitioned mixture model (SS-RPMM) approach to identify novel blood DNA methylation markers of HNSCC using genome-wide methylation array data for peripheral blood samples from 92 HNSCC cases and 92 cancer-free control subjects. To assess the performance of the resultant markers, we constructed receiver operating characteristic (RJC) curves and calculated the corresponding area under the curve (AUC). Cases and controls were best differentiated by a methylation profile of six CpG loci (associated with FGD4, SERPINF1, WDR39, IL27, HYAL2 and PLEKHA6), with an AUC of 0.73 (95% CI: 0.62–0.82). After adjustment for subject age, gender, smoking, alcohol consumption and HPV16 serostatus, the AUC increased to 0.85 (95% CI: 0.76–0.92). We have identified a novel blood-based methylation profile that is indicative of HNSCC with a high degree of accuracy. This profile demonstrates the potential of DNA methylation measured in blood for development of non-invasive applications for detection of head and neck cancer. PMID:22430805

  8. Infants' Peripheral Blood Lymphocyte Composition Reflects Both Maternal and Post-Natal Infection with Plasmodium falciparum

    PubMed Central

    Ibitokou, Samad; Vianou, Bertin; Houngbegnon, Parfait; Ezinmegnon, Sem; Borgella, Sophie; Akplogan, Carine; Cottrell, Gilles; Varani, Stefania; Massougbodji, Achille; Moutairou, Kabirou; Troye-Blomberg, Marita; Deloron, Philippe; Luty, Adrian J. F.; Fievet, Nadine

    2015-01-01

    Maternal parasitoses modulate fetal immune development, manifesting as altered cellular immunological activity in cord blood that may be linked to enhanced susceptibility to infections in early life. Plasmodium falciparum typifies such infections, with distinct placental infection-related changes in cord blood exemplified by expanded populations of parasite antigen-specific regulatory T cells. Here we addressed whether such early-onset cellular immunological alterations persist through infancy. Specifically, in order to assess the potential impacts of P. falciparum infections either during pregnancy or during infancy, we quantified lymphocyte subsets in cord blood and in infants' peripheral blood during the first year of life. The principal age-related changes observed, independent of infection status, concerned decreases in the frequencies of CD4+, NKdim and NKT cells, whilst CD8+, Treg and Teff cells' frequencies increased from birth to 12 months of age. P. falciparum infections present at delivery, but not those earlier in gestation, were associated with increased frequencies of Treg and CD8+ T cells but fewer CD4+ and NKT cells during infancy, thus accentuating the observed age-related patterns. Overall, P. falciparum infections arising during infancy were associated with a reversal of the trends associated with maternal infection i.e. with more CD4+ cells, with fewer Treg and CD8+ cells. We conclude that maternal P. falciparum infection at delivery has significant and, in some cases, year-long effects on the composition of infants' peripheral blood lymphocyte populations. Those effects are superimposed on separate and independent age- as well as infant infection-related alterations that, respectively, either match or run counter to them. PMID:26580401

  9. Direct polymerase chain reaction from blood and tissue samples for rapid diagnosis of bovine leukemia virus infection.

    PubMed

    Nishimori, Asami; Konnai, Satoru; Ikebuchi, Ryoyo; Okagawa, Tomohiro; Nakahara, Ayako; Murata, Shiro; Ohashi, Kazuhiko

    2016-06-01

    Bovine leukemia virus (BLV) infection induces bovine leukemia in cattle and causes significant financial harm to farmers and farm management. There is no effective therapy or vaccine; thus, the diagnosis and elimination of BLV-infected cattle are the most effective method to eradicate the infection. Clinical veterinarians need a simpler and more rapid method of diagnosing infection, because both nested polymerase chain reaction (PCR) and real-time PCR are labor intensive, time-consuming, and require specialized molecular biology techniques and expensive equipment. In this study, we describe a novel PCR method for amplifying the BLV provirus from whole blood, thus eliminating the need for DNA extraction. Although the sensitivity of PCR directly from whole blood (PCR-DB) samples as measured in bovine blood containing BLV-infected cell lines was lower than that of nested PCR, the PCR-DB technique showed high specificity and reproducibility. Among 225 clinical samples, 49 samples were positive by nested PCR, and 37 samples were positive by PCR-DB. There were no false positive samples; thus, PCR-DB sensitivity and specificity were 75.51% and 100%, respectively. However, the provirus loads of the samples detected by nested PCR and not PCR-DB were quite low. Moreover, PCR-DB also stably amplified the BLV provirus from tumor tissue samples. PCR-DB method exhibited good reproducibility and excellent specificity and is suitable for screening of thousands of cattle, thus serving as a viable alternative to nested PCR and real-time PCR.

  10. Expression of extracellular calcium (Ca2+o)-sensing receptor in human peripheral blood monocytes

    NASA Technical Reports Server (NTRS)

    Yamaguchi, T.; Olozak, I.; Chattopadhyay, N.; Butters, R. R.; Kifor, O.; Scadden, D. T.; Brown, E. M.; O'Malley, B. W. (Principal Investigator)

    1998-01-01

    The calcium-sensing receptor (CaR) is a G protein-coupled receptor playing key roles in extracellular calcium ion (Ca2+o) homeostasis in parathyroid gland and kidney. Macrophage-like mononuclear cells appear at sites of osteoclastic bone resorption during bone turnover and may play a role in the "reversal" phase of skeletal remodeling that follows osteoclastic resorption and precedes osteoblastic bone formation. Bone resorption produces substantial local increases in Ca2+o that could provide a signal for such mononuclear cells present locally within the bone marrow microenvironment. Indeed, previous studies by other investigators have shown that raising Ca2+o either in vivo or in vitro stimulated the release of interleukin-6 (IL-6) from human peripheral blood monocytes, suggesting that these cells express a Ca2+o-sensing mechanism. In these earlier studies, however, the use of reverse transcription-polymerase chain reaction (RT-PCR) failed to detect transcripts for the CaR previously cloned from parathyroid and kidney in peripheral blood monocytes. Since we recently found that non-specific esterase-positive, putative monocytes isolated from murine bone marrow express the CaR, we reevaluated the expression of this receptor in human peripheral blood monocytes. Immunocytochemistry, flow cytometry, and Western blot analysis, performed using a polyclonal antiserum specific for the CaR, detected CaR protein in human monocytes. In addition, the use of RT-PCR with CaR-specific primers, followed by nucleotide sequencing of the amplified products, identified CaR transcripts in the cells. Therefore, taken together, our data show that human peripheral blood monocytes possess both CaR protein and mRNA very similar if not identical to those expressed in parathyroid and kidney that could mediate the previously described, direct effects of Ca2+o on these cells. Furthermore, since mononuclear cells isolated from bone marrow also express the CaR, the latter might play some role in

  11. The effects of lipid A on gamma-irradiated human peripheral blood lymphocytes in vitro

    NASA Astrophysics Data System (ADS)

    Dubničková, M.; Kuzmina, E. A.; Chausov, V. N.; Ravnachka, I.; Boreyko, A. V.; Krasavin, E. A.

    2016-03-01

    The modulatory effects of lipid A (diphosphoryl lipid A (DLA) and monophosphoryl lipid A (MLA)) on apoptosis induction and DNA structure damage (single and double-strand breaks (SSBs and DSBs, respectively)) in peripheral human blood lymphocytes are studied for 60Co gamma-irradiation. It is shown that in the presence of these agents the amount of apoptotic cells increases compared with the irradiated control samples. The effect is most strongly pronounced for DLA. In its presence, a significant increase is observed in the number of radiation-induced DNA SSBs and DSBs. Possible mechanisms are discussed of the modifying influence of the used agents on radiation-induced cell reactions

  12. Unexpected postmortem diagnosis of Acanthamoeba meningoencephalitis following allogeneic peripheral blood stem cell transplantation.

    PubMed

    Pemán, J; Jarque, I; Frasquet, J; Alberola, C; Salavert, M; Sanz, J; Gomila, B; Esteban, G

    2008-07-01

    Meningoencephalitis caused by pathogenic free-living amebas is usually fatal. Only a few cases of Acanthamoeba meningoencephalitis, diagnosed at autopsy, have been reported following hematopoietic stem cell transplantation. We here report a case of Acanthamoeba meningoencephalitis following allogeneic peripheral blood stem cell transplantation with rapidly evolving neurologic symptoms that remained unexplained. Magnetic resonance imaging failed to show brain lesions and cerebrospinal fluid was negative for microbiological cultures. Definite diagnosis was an unexpected autopsy finding. As overall and teaching hospital autopsy rates are declining worldwide, we must emphasize the need of autopsy exams if we want to improve our knowledge as the best way to care for our patients.

  13. Interferon-inducible gene expression signature in peripheral blood cells of patients with severe lupus

    PubMed Central

    Baechler, Emily C.; Batliwalla, Franak M.; Karypis, George; Gaffney, Patrick M.; Ortmann, Ward A.; Espe, Karl J.; Shark, Katherine B.; Grande, William J.; Hughes, Karis M.; Kapur, Vivek; Gregersen, Peter K.; Behrens, Timothy W.

    2003-01-01

    Systemic lupus erythematosus (SLE) is a complex, inflammatory autoimmune disease that affects multiple organ systems. We used global gene expression profiling of peripheral blood mononuclear cells to identify distinct patterns of gene expression that distinguish most SLE patients from healthy controls. Strikingly, about half of the patients studied showed dysregulated expression of genes in the IFN pathway. Furthermore, this IFN gene expression “signature” served as a marker for more severe disease involving the kidneys, hematopoetic cells, and/or the central nervous system. These results provide insights into the genetic pathways underlying SLE, and identify a subgroup of patients who may benefit from therapies targeting the IFN pathway. PMID:12604793

  14. Expression of extracellular calcium (Ca2+o)-sensing receptor in human peripheral blood monocytes

    NASA Technical Reports Server (NTRS)

    Yamaguchi, T.; Olozak, I.; Chattopadhyay, N.; Butters, R. R.; Kifor, O.; Scadden, D. T.; Brown, E. M.; O'Malley, B. W. (Principal Investigator)

    1998-01-01

    The calcium-sensing receptor (CaR) is a G protein-coupled receptor playing key roles in extracellular calcium ion (Ca2+o) homeostasis in parathyroid gland and kidney. Macrophage-like mononuclear cells appear at sites of osteoclastic bone resorption during bone turnover and may play a role in the "reversal" phase of skeletal remodeling that follows osteoclastic resorption and precedes osteoblastic bone formation. Bone resorption produces substantial local increases in Ca2+o that could provide a signal for such mononuclear cells present locally within the bone marrow microenvironment. Indeed, previous studies by other investigators have shown that raising Ca2+o either in vivo or in vitro stimulated the release of interleukin-6 (IL-6) from human peripheral blood monocytes, suggesting that these cells express a Ca2+o-sensing mechanism. In these earlier studies, however, the use of reverse transcription-polymerase chain reaction (RT-PCR) failed to detect transcripts for the CaR previously cloned from parathyroid and kidney in peripheral blood monocytes. Since we recently found that non-specific esterase-positive, putative monocytes isolated from murine bone marrow express the CaR, we reevaluated the expression of this receptor in human peripheral blood monocytes. Immunocytochemistry, flow cytometry, and Western blot analysis, performed using a polyclonal antiserum specific for the CaR, detected CaR protein in human monocytes. In addition, the use of RT-PCR with CaR-specific primers, followed by nucleotide sequencing of the amplified products, identified CaR transcripts in the cells. Therefore, taken together, our data show that human peripheral blood monocytes possess both CaR protein and mRNA very similar if not identical to those expressed in parathyroid and kidney that could mediate the previously described, direct effects of Ca2+o on these cells. Furthermore, since mononuclear cells isolated from bone marrow also express the CaR, the latter might play some role in

  15. Interferon-inducible gene expression signature in peripheral blood cells of patients with severe lupus.

    PubMed

    Baechler, Emily C; Batliwalla, Franak M; Karypis, George; Gaffney, Patrick M; Ortmann, Ward A; Espe, Karl J; Shark, Katherine B; Grande, William J; Hughes, Karis M; Kapur, Vivek; Gregersen, Peter K; Behrens, Timothy W

    2003-03-04

    Systemic lupus erythematosus (SLE) is a complex, inflammatory autoimmune disease that affects multiple organ systems. We used global gene expression profiling of peripheral blood mononuclear cells to identify distinct patterns of gene expression that distinguish most SLE patients from healthy controls. Strikingly, about half of the patients studied showed dysregulated expression of genes in the IFN pathway. Furthermore, this IFN gene expression "signature" served as a marker for more severe disease involving the kidneys, hematopoetic cells, and/or the central nervous system. These results provide insights into the genetic pathways underlying SLE, and identify a subgroup of patients who may benefit from therapies targeting the IFN pathway.

  16. Almond Skin Inhibits HSV-2 Replication in Peripheral Blood Mononuclear Cells by Modulating the Cytokine Network.

    PubMed

    Arena, Adriana; Bisignano, Carlo; Stassi, Giovanna; Filocamo, Angela; Mandalari, Giuseppina

    2015-05-15

    We have investigated the effect of almond skin extracts on the production of pro-inflammatory and anti-inflammatory cytokines in human peripheral blood mononuclear cells (PBMCs). PBMCs were either infected or not by herpes simplex virus type 2 (HSV-2), with and without prior treatment with almond skin extracts. Production of IL-17 induced by HSV-2 was inhibited by natural skins (NS) treatment. NS triggered PBMC in releasing IFN-α, IFN-γ and IL-4 in cellular supernatants. These results may explain the antiviral potential of almond skins.

  17. [Structure of peripheral blood platelets surface in patients with amyotrophic lateral sclerosis and multiple sclerosis].

    PubMed

    Kiktenko, A I; Zlobina, G P; Brusov, O S; Zakharova, M N

    2005-01-01

    Using scanning electronic microscopy of peripheral blood platelets in 32 patients with amyotrophic lateral sclerosis (ALS) and 6 patients with multiple sclerosis (MS), platelets activation, emerging in pseudopodiums formation, aggregation and lysis, was found. Platelets activation was more pronounced in ALS than in MS. It was most markedly seen for quantitative evaluation of the above traits. The features of platelet activation found in the study support the evidence for considering ALS as a systemic disorder being characterized by a broad spectrum of alterations of biological processes.

  18. Similar blood-borne DNA methylation alterations in cancer and inflammatory diseases determined by subpopulation shifts in peripheral leukocytes.

    PubMed

    Li, H; Zheng, T; Chen, B; Hong, G; Zhang, W; Shi, T; Li, S; Ao, L; Wang, C; Guo, Z

    2014-07-29

    Although many DNA methylation (DNAm) alterations observed in peripheral whole blood/leukocytes and serum have been considered as potential diagnostic markers for cancer, their origin and their specificity for cancer (e.g., vs inflammatory diseases) remain unclear. From publicly available datasets, we identified changes in the methylation of blood-borne DNA for multiple cancers and inflammatory diseases. We compared the identified changes with DNAm difference between myeloid and lymphoid cells extracted from two datasets. At least 94.7% of the differentially methylated DNA loci (DM loci) observed in peripheral whole blood/leukocytes and serum of cancer patients overlapped with DM loci that distinguish between myeloid and lymphoid cells and >99.9% of the overlapped DM loci had consistent alteration states (hyper- or hypomethylation) in cancer samples compared to normal controls with those in myeloid cells compared to lymphoid cells (binomial test, P-value <2.2 × 10(-16)). Similar results were observed for DM loci in peripheral whole blood/leukocytes in patients with rheumatoid arthritis or inflammatory bowel diseases. The direct comparison between DM loci observed in the peripheral whole blood/leukocytes of patients with inflammatory diseases and DM loci observed in the peripheral whole blood of patients with cancer showed that DM loci detected from cancer and inflammatory diseases also had significantly consistent alteration states (binomial test, P-value <2.2 × 10(-16)). DNAm changes observed in the peripheral whole blood/leukocytes and serum of cancer patients and in the peripheral whole blood/leukocytes of inflammatory disease patients are predominantly determined by the increase of myeloid cells and the decrease of lymphoid cells under the disease conditions, in the sense that their alteration states in disease samples compared to normal controls mainly reflect the DNAm difference between myeloid and lymphoid cells. These analyses highlight the importance of

  19. Variety of RNAs in Peripheral Blood Cells, Plasma, and Plasma Fractions

    PubMed Central

    Kuligina, Elena V.; Bariakin, Dmitry N.; Kozlov, Vadim V.; Richter, Vladimir A.; Semenov, Dmitry V.

    2017-01-01

    Human peripheral blood contains RNA in cells and in extracellular membrane vesicles, microvesicles and exosomes, as well as in cell-free ribonucleoproteins. Circulating mRNAs and noncoding RNAs, being internalized, possess the ability to modulate vital processes in recipient cells. In this study, with SOLiD sequencing technology, we performed identification, classification, and quantification of RNAs from blood fractions: cells, plasma, plasma vesicles pelleted at 16,000g and 160,000g, and vesicle-depleted plasma supernatant of healthy donors and non-small cell lung cancer (NSCLC) patients. It was determined that 16,000g blood plasma vesicles were enriched with cell-free mitochondria and with a set of mitochondrial RNAs. The variable RNA set of blood plasma 160,000g pellets reflected the prominent contribution of U1, U5, and U6 small nuclear RNAs' fragments and at the same time was characterized by a remarkable depletion of small nucleolar RNAs. Besides microRNAs, the variety of fragments of mRNAs and snoRNAs dominated in the set of circulating RNAs differentially expressed in blood fractions of NSCLC patients. Taken together, our data emphasize that not only extracellular microRNAs but also circulating fragments of messenger and small nuclear/nucleolar RNAs represent prominent classes of circulating regulatory ncRNAs as well as promising circulating biomarkers for the development of disease diagnostic approaches. PMID:28127559

  20. Study of plasma-induced peripheral blood mononuclear cells survival using Fourier transform infrared microspectroscopy.

    PubMed

    Sahu, Ranjit K; Salman, Ahmad; Mordechai, Shaul; Manor, Esther

    2013-11-01

    Components present in the acellular fraction of blood influence the blood cell survival and function and the response to biotic and abiotic factors. Human plasma and sera have been used as therapeutic agents and are known to increase cell survival. White blood cells in normal blood are exposed to plasma components in vivo, but the effect of such plasma components in vitro on adherent peripheral blood mononuclear cells (PBMCs) that includes monocytes has not been fully investigated. We cultured human PBMCs with autologous plasma and observed structural variation due to plasma addition in PBMCs along with increased cell survival. Light microscopy of the cells showed increased granularity in plasma-treated cells. Fourier transform infrared (FTIR) spectroscopy was used to elucidate the possible mechanism by studying the changes in the biochemical composition of the cells that explained the observations. FTIR spectroscopy of plasma-treated cells show altered spectral pattern in the mid-IR region, indicating increased phospholipid levels. Heat-stable components in the plasma possibly increase the differentiation of PBMCs, as evident by increased phospholipid metabolism. The data suggest that plasma-stimulated membrane biogenesis may contribute to PBMC survival by inducing them to differentiate into antigen presenting cells (APCs) like macrophages and dendritic cells.

  1. Variety of RNAs in Peripheral Blood Cells, Plasma, and Plasma Fractions.

    PubMed

    Savelyeva, Anna V; Kuligina, Elena V; Bariakin, Dmitry N; Kozlov, Vadim V; Ryabchikova, Elena I; Richter, Vladimir A; Semenov, Dmitry V

    2017-01-01

    Human peripheral blood contains RNA in cells and in extracellular membrane vesicles, microvesicles and exosomes, as well as in cell-free ribonucleoproteins. Circulating mRNAs and noncoding RNAs, being internalized, possess the ability to modulate vital processes in recipient cells. In this study, with SOLiD sequencing technology, we performed identification, classification, and quantification of RNAs from blood fractions: cells, plasma, plasma vesicles pelleted at 16,000g and 160,000g, and vesicle-depleted plasma supernatant of healthy donors and non-small cell lung cancer (NSCLC) patients. It was determined that 16,000g blood plasma vesicles were enriched with cell-free mitochondria and with a set of mitochondrial RNAs. The variable RNA set of blood plasma 160,000g pellets reflected the prominent contribution of U1, U5, and U6 small nuclear RNAs' fragments and at the same time was characterized by a remarkable depletion of small nucleolar RNAs. Besides microRNAs, the variety of fragments of mRNAs and snoRNAs dominated in the set of circulating RNAs differentially expressed in blood fractions of NSCLC patients. Taken together, our data emphasize that not only extracellular microRNAs but also circulating fragments of messenger and small nuclear/nucleolar RNAs represent prominent classes of circulating regulatory ncRNAs as well as promising circulating biomarkers for the development of disease diagnostic approaches.

  2. Detection of circulating tumor cells from cryopreserved human sarcoma peripheral blood mononuclear cells.

    PubMed

    Li, Heming; Meng, Qing H; Noh, Hyangsoon; Batth, Izhar Singh; Somaiah, Neeta; Torres, Keila E; Xia, Xueqing; Wang, Ruoyu; Li, Shulin

    2017-09-10

    Circulating tumor cells (CTCs) enter the vasculature or lymphatic system after shedding from the primary tumor. CTCs may serve as "seed" cells for tumor metastasis. The utility of CTCs in clinical applications for sarcoma is not fully investigated, partly owing to the necessity for fresh blood samples and the lack of a CTC-specific antibody. To overcome these drawbacks, we developed a technique for sarcoma CTCs capture and detection using cryopreserved peripheral blood mononuclear cells (PBMCs) and our proprietary cell-surface vimentin (CSV) antibody 84-1, which is specific to tumor cells. This technique was validated by sarcoma cell spiking assay, matched CTCs comparison between fresh and cryopreserved PBMCs, and independent tumor markers in multiple types of sarcoma patient blood samples. The reproducibility was maximized when cryopreserved PBMCs were prepared from fresh blood samples within 2 h of the blood draw. In summary, as far as we are aware, ours is the first report to capture and detect CTCs from cryopreserved PBMCs. Further validation in other types of tumor may help boost the feasibility and utility of CTC-based diagnosis in a centralized laboratory. Copyright © 2017 Elsevier B.V. All rights reserved.

  3. Quantification of peripheral and central blood pressure variability using a time-frequency method.

    PubMed

    Kouchaki, Z; Butlin, M; Qasem, A; Avolio, A P; Kouchaki, Z; Butlin, M; Qasem, A; Avolio, A P; Kouchaki, Z; Avolio, A P; Butlin, M; Qasem, A

    2016-08-01

    Systolic blood pressure variability (BPV) is associated with cardiovascular events. As the beat-to-beat variation of blood pressure is due to interaction of several cardiovascular control systems operating with different response times, assessment of BPV by spectral analysis using the continuous measurement of arterial pressure in the finger is used to differentiate the contribution of these systems in regulating blood pressure. However, as baroreceptors are centrally located, this study considered applying a continuous aortic pressure signal estimated noninvasively from finger pressure for assessment of systolic BPV by a time-frequency method using Short Time Fourier Transform (STFT). The average ratio of low frequency and high frequency power band (LFPB/HFPB) was computed by time-frequency decomposition of peripheral systolic pressure (pSBP) and derived central aortic systolic blood pressure (cSBP) in 30 healthy subjects (25-62 years) as a marker of balance between cardiovascular control systems contributing in low and high frequency blood pressure variability. The results showed that the BPV assessed from finger pressure (pBPV) overestimated the BPV values compared to that assessed from central aortic pressure (cBPV) for identical cardiac cycles (P<;0.001), with the overestimation being greater at higher power.

  4. [Artificial control of blood-nerve barrier: a novel therapeutic approach to peripheral neuropathies].

    PubMed

    Kanda, Takashi

    2011-11-01

    Blood-nerve barrier (BNB) is a "Janus-faced" structure for the peripheral nerve parenchyma. Healthy BNB may contribute to stabilize the internal milleu of peripheral nervous system (PNS) and to stop the entrance of toxic substances and harmful leukocytes into nerve parenchyma. On the other hand, healthy BNB may sometimes be a drawback because the peripheral nerve parenchyma cannot receive enough amount of nutrients and growth factors and cannot excrete toxic substances into systemic circulation because of its presence. Here we present a future therapeutic strategy to control BNB function, based on the basic knowledge acquired from recently developed human immortalized cell lines of BNB origin. If we can artificially regulate the BNB permeability and the expression of adhesion molecules on the surface of BNB-forming endothelial cells, and stop the entrance of toxic substances as well as pathogenic leukocytes into PNS parenchyma, the treatment of inflammatory neuropathies may make great progresses. For hereditary, metabolic and ischemic neuropathies, the promotion of the entrance of growth factors into PNS parenchyma and of the excretion of toxic substances should powerfully encourage the regeneration of axons.

  5. Menstrual blood closely resembles the uterine immune micro-environment and is clearly distinct from peripheral blood.

    PubMed

    van der Molen, R G; Schutten, J H F; van Cranenbroek, B; ter Meer, M; Donckers, J; Scholten, R R; van der Heijden, O W H; Spaanderman, M E A; Joosten, I

    2014-02-01

    Is menstrual blood a suitable source of endometrial derived lymphocytes? Mononuclear cells isolated from menstrual samples (menstrual blood mononuclear cells (MMC)) are clearly distinct from peripheral blood mononuclear cells (PBMC) and show a strong resemblance with biopsy-derived endometrial mononuclear cells. A critical event in the onset of pregnancy is the implantation of the embryo in the uterine wall. The immune cell composition in the endometrium at the time of implantation is considered pivotal for success. Despite advancing knowledge on the composition of the immune cell population in the uterus, the role of endometrial immune cells in reproductive disorders is still not fully resolved, mainly due to the fact that this type of research requires invasive techniques. Here, we collected menstrual fluid and validated this unique non-invasive technique to obtain and study the endometrium-derived immune cells which would be present around the time of implantation. Five healthy non-pregnant females with regular menstruation cycles and not using oral contraceptives collected their menstrual blood using a menstrual cup in five consecutive cycles. Sampling took place over the first 3 days of menses, with 12 h intervals. Peripheral blood samples, taken before and after each menstruation, were obtained for comparative analysis. MMC and PBMC samples were characterized for the different lymphocyte subsets by flow cytometry, with emphasis on NK cells and T cells. Next, the functional capacity of the MMC-derived NK cells was determined by measuring intracellular production of IFN-γ, granzyme B and perforin after culture in the presence of IL-2 and IL-15. In support of their endometrial origin, MMC samples contained the typical composition of mononuclear cells expected of endometrial tissue, were phenotypically similar to the reported phenotype for biopsy-derived endometrial cells, and were distinct from PBMC. Increased percentages of NK cells and decreased percentages

  6. Crystalglobulinemia with fulminant course with cylinder-like bodies on peripheral blood smear.

    PubMed

    Kawaguchi, Takeshi; Kariya, Yumi; Matsuda, Motohiro; Kubo, Kazuyoshi; Miyauchi, Syunichi; Kusumoto, Norio; Ueno, Shiro; Takajo, Ichiro; Nagatomo, Yasuhiro; Tahira, Yuki; Yamamoto, Syoko; Yorita, Kenji; Kataoka, Hiroaki; Okayama, Akihiko

    2014-01-01

    A 63-year-old woman presented to our hospital with fever, purpura and pain in both legs and died 4 days after admission. Her blood smear and skin biopsy showed cylinder-like bodies (20×120 μm). She was diagnosed to have monoclonal gammopathy (IgG, lambda type). An autopsy revealed cylinder-like bodies in the vasculature of various organs. We noted a proliferation of atypical plasma cells in her bone marrow, suggesting pre-existing myeloma. Crystalglobulinemia is a rare manifestation of hypergammaglobulinemia that can cause multiple embolisms of the small vessels, and this resulted in the patient's fulminant course. The identification of cylinder-like bodies in the peripheral blood may help in reaching a diagnosis in such cases.

  7. Isolation of Mesenchymal Stromal Cells From Peripheral Blood of ST Elevation Myocardial Infarction Patients.

    PubMed

    Pieper, Ina Laura; Smith, Rachel; Bishop, Joanna C; Aldalati, Omar; Chase, Alex J; Morgan, Gareth; Thornton, Catherine A

    2017-02-28

    Bone marrow mesenchymal stromal cells (MSCs) have shown therapeutic potential in the treatment of myocardial infarction patients. However, bone marrow requires invasive harvesting techniques. Therefore, the aim was to carry out a feasibility study of using autologous peripheral blood (PB) as a source for MSCs and platelet lysate (PL), a potential novel therapeutic intervention in acute ST elevation myocardial infarction (STEMI) patients. Autologous PL and MSCs were prepared from STEMI patient and healthy control blood. MSCs were analyzed by trilineage differentiation and flow cytometry. PB MSCs were isolated from 83% of patients (n = 6) but not from controls. The use of PL was feasible in the first passage but not in subsequent ones due to volume. To conclude, PB is a promising alternative to bone marrow. It negates the need for invasive harvesting techniques, and reduces hemorrhagic risk in this patient population routinely managed with anticoagulant and antiplatelet agents.

  8. Junin virus replication in peripheral blood mononuclear cells of patients with Argentine haemorrhagic fever.

    PubMed

    Ambrosio, M; Vallejos, A; Saavedra, C; Maiztegui, J I

    1990-02-01

    To study the relationship of Junin virus (JV) to populations of peripheral blood mononuclear cells (PBMC) from patients with Argentine Haemorrhagic Fever (AHF), blood samples were obtained during the acute period of disease and cultured as total, adherent, and non-adherent cell populations. JV was sequentially sought in these cell populations by using an Infectious Centre (IC) assay, whereas free JV in the supernatants was evaluated by plaque formation. IC were obtained in cultures of total PBMC from 8 out of 19 patients. Maximum numbers of IC showed high variation among patients, ranging from 3 to 410 IC per 10(6) viable PBMC. In contrast, IC were sporadically demonstrated in the non-adherent cell population. The release of JV into culture supernatants was detected only in total PBMC cultures, thus in the presence of macrophages. These results demonstrate that circulating monocytes (macrophages) are targets for JV replication contributing to the viral spread in the acute phase of AHF.

  9. [Peripheral blood of children exposed to radiation as a consequence of the Chernobyl AES accident].

    PubMed

    Torubarova, D A; Kovalev, G I

    1991-01-01

    As many as 103 children exposed to minor doses of radiation after the Chernobyl accident were examined for peripheral blood morphology. Statistical and individual analysis did not reveal any pathological alterations on the part of the hemograms of the radiated children. At the same time they manifested certain deviations in the form of leukopenia, lymphopenia and neutropenia, suggesting the action produced by radiation factor. On the whole, these alterations were characterized as adaptation ones of multifactorial genesis. The data obtained support an assumption that the changes in the quantitative composition of blood exposed to radiation in the doses not exceeding the maximal permissible limits were not remarkable, occurring within the physiological boundaries and could be detected only during observations made over time. Emphasis is laid on the necessity of further monitoring of the hemopoietic system of the radiated children according to the current principles of dispensary observation.

  10. [Reference intervals for peripheral blood lymphocyte subsets in healthy adults in Lima, Peru].

    PubMed

    Cóndor, José M; Álvarez, Marco; Cano, Luis; Matos, Edgar; Leiva, Christian; Paredes, José A

    2013-04-01

    In order to establish the reference intervals (RIs) of peripheral blood lymphocyte subsets (PBL) in healthy adults in Lima (Peru), a cross-sectional study was conducted among blood donors taken in between 2011 and 2012. Based on the criteria obtained from the guidelines of the Clinical and Laboratory Standards Institute (CLSI C28-A3), 318 samples were processed, 61.9% (197/318) coming from male donors. For PBL count, a flow cytometer with a simple platform was used. The RIs are established for each PBL in adults based on sex with their respective reference limits and 90% confidence intervals. Differences were found in CD3+ percentage counts (p=0.001) and in CD3-CD56+ absolute (p=0.003) and percentage counts (p?0.001). The RIs found are different to those described in studies conducted in other countries due to the characteristics of the population and the study model.

  11. [The effect of space flight factors on the peripheral blood in the newt Pleurodeles waltlii].

    PubMed

    Domaratskaia, E I; Mirchurina, T V; Nikonova, T M; Khrushchov, N G

    1994-01-01

    The effects of space flight factors (SFF) on the peripheral blood in Pleurodeles waltlii were assessed after 12-day flight on board of the biosatellite "Kosmos-2229". These animals were also used to study regeneration of the limb, tail and lens. The corresponding control groups of animals allowed to distinguish between the effects of the operation, non-specific and specific SFFs: (1) basal control-operated animals; (2) synchronous control-operated animals kept on the Earth under the same conditions as the flight group, and (3) intact animals. It has been shown that the relative content of neutrophils (mostly, young forms) increased and the proportion of lymphocytes and eosinophils decreased under the influence of SFFs, while the capacity of blood cells for DNA synthesis was not affected. A conclusion has been drawn that the Spanish newts can be used for adequate studies of the SFF effects on the hemopoietic tissue.

  12. Detection of Intracellular Factor VIII Protein in Peripheral Blood Mononuclear Cells by Flow Cytometry

    PubMed Central

    Pandey, Gouri Shankar; Tseng, Sandra C.; Howard, Tom E.; Sauna, Zuben E.

    2013-01-01

    Flow cytometry is widely used in cancer research for diagnosis, detection of minimal residual disease, as well as immune monitoring and profiling following immunotherapy. Detection of specific host proteins for diagnosis predominantly uses quantitative PCR and western blotting assays. In this study, we optimized a flow cytometry-based detection assay for Factor VIII protein in peripheral blood mononuclear cells (PBMCs). An indirect intracellular staining (ICS) method was standardized using monoclonal antibodies to different domains of human Factor VIII protein. The FVIII protein expression level was estimated by calculating the mean and median fluorescence intensities (MFI) values for each monoclonal antibody. ICS staining of transiently transfected cell lines supported the method's specificity. Intracellular FVIII protein expression was also detected by the monoclonal antibodies used in the study in PBMCs of five blood donors. In summary, our data suggest that intracellular FVIII detection in PBMCs of hemophilia A patients can be a rapid and reliable method to detect intracellular FVIII levels. PMID:23555096

  13. DNA damage in peripheral blood mononuclear cells and neutrophils of dairy cows during the transition period.

    PubMed

    Tharwat, M; Endoh, D; Oikawa, S

    2012-01-01

    This study was designed to investigate the apoptotic process in peripheral blood mononuclear cells (PBMC) and polymorphonuclear neutrophil leukocytes (PMN) in dairy cattle during the transition period. Blood samples were collected from 4 dairy cattle at 3 weeks before the expected parturition (wk -3), parturition (wk 0) and 3 weeks after parturition (wk +3). The DNA damage of PBMC and PMN was evaluated based on the comet assay using visual scoring (arbitrary units). Undamaged DNA remained within the core (score 0) and the broken DNA migrated from the core towards the anode forming the tail of a comet (scores 1-4). Significantly higher scores in PBMC at wk 0 and wk +3 were observed compared with those in PMN although there were no significant changes of scores in either cell type during the experimental period. It is suggested that the apoptotic rate of PBMC is accelerated compared with that of PMC during the transition period.

  14. Flow cytometric enumeration of absolute lymphocyte number in peripheral blood using two parameters of light scatter.

    PubMed

    Smart, Y C; Cox, J; Murphy, B; Enno, A; Burton, R C

    1985-03-01

    A method was developed to measure the absolute lymphocyte count (ALC) of whole blood using the Spectrum III automated flow cytometer. Ninety-nine samples of human peripheral blood were analysed on the Spectrum and the Coulter Counter S Plus II, to allow for comparison of the two machines. Regression analysis was used to test the extent of agreement between the sets of measurements on the two machines. The results demonstrated that the slope of the regression line was not significantly different from one, indicating a high level of correlation between Spectrum and Coulter ALC's. However, the mean difference between Coulter and Spectrum ALC's was not equal to zero, with the Spectrum giving counts approximately 10% lower than those of the Coulter machine. This is attributed to the different ways by which the two machines define a lymphocyte, the Spectrum III by two parameters of light scatter and the Coulter S Plus II by the single parameter of cell volume.

  15. Massive delayed hemolysis following peripheral blood stem cell transplantation with minor ABO incompatibility.

    PubMed

    Laurencet, F M; Samii, K; Bressoud, A; Tajeddin, M; Easton, J; Stelling, M J; Chapuis, B

    1997-06-01

    After hematopoietic stem cell transplantation, delayed immune hemolysis may occur when donor-derived B lymphocytes carried with the graft produce immune antibodies against the recipient's incompatible red cells. We report the occurrence of this syndrome in the context of minor blood group incompatibility between donor and recipient after peripheral blood stem cell (PBSC) transplantation. On day 12 post-transplant there was abrupt onset of hemolysis necessitating supportive treatment with hydration and transfusions. Because, as compared to bone marrow, PBSC grafts are enriched with lymphocytes, more frequent and intense delayed immune hemolysis may be anticipated when using PBSC. This complication is described most often when cyclosporine alone is used for immunosuppression following the graft. The addition of methotrexate, which with CyA forms the classic regimen for the prevention of graft-vs-host disease, may diminish the frequence and severity of this adverse reaction.

  16. Time-resolved fluorescence monitoring of cholesterol in peripheral blood mononuclear cells

    NASA Astrophysics Data System (ADS)

    Martinakova, Z.; Horilova, J.; Lajdova, I.; Marcek Chorvatova, A.

    2014-12-01

    Precise evaluation of intracellular cholesterol distribution is crucial for improving diagnostics of diseased states associated with cholesterol alteration. Time-resolved fluorescence techniques are tested for non-invasive investigation of cholesterol in living cells. Fluorescent probe NBD attached to cholesterol was employed to evaluate cholesterol distribution in peripheral blood mononuclear cells (PBMC) isolated from the human blood. Fluorescence Lifetime Imaging Microscopy (FLIM) was successfully applied to simultaneously monitor the spatial distribution and the timeresolved characteristics of the NBD-cholesterol fluorescence in PBMC. Gathered data are the first step in the development of a new perspective non-invasive diagnostic method for evaluation of cholesterol modifications in diseases associated with disorders of lipid metabolism.

  17. Blood borne hormones in a cross-talk between peripheral and brain mechanisms regulating blood pressure, the role of circumventricular organs.

    PubMed

    Ufnal, Marcin; Skrzypecki, Janusz

    2014-04-01

    Accumulating evidence suggests that blood borne hormones modulate brain mechanisms regulating blood pressure. This appears to be mediated by the circumventricular organs which are located in the walls of the brain ventricular system and lack the blood-brain barrier. Recent evidence shows that neurons of the circumventricular organs express receptors for the majority of cardiovascular hormones. Intracerebroventricular infusions of hormones and their antagonists is one approach to evaluate the influence of blood borne hormones on the neural mechanisms regulating arterial blood pressure. Interestingly, there is no clear correlation between peripheral and central effects of cardiovascular hormones. For example, angiotensin II increases blood pressure acting peripherally and centrally, whereas peripherally acting pressor catecholamines decrease blood pressure when infused intracerebroventricularly. The physiological role of such dual hemodynamic responses has not yet been clarified. In the paper we review studies on hemodynamic effects of catecholamines, neuropeptide Y, angiotensin II, aldosterone, natriuretic peptides, endothelins, histamine and bradykinin in the context of their role in a cross-talk between peripheral and brain mechanisms involved in the regulation of arterial blood pressure. Copyright © 2014 Elsevier Ltd. All rights reserved.

  18. Development of subsequent bloodstream infection in patients with positive Hickman catheter blood cultures and negative peripheral blood cultures.

    PubMed

    Park, Ki-Ho; Cho, Oh-Hyun; Lee, Sang-Oh; Choi, Sang-Ho; Kim, Yang Soo; Woo, Jun Hee; Kim, Mi-Na; Kim, Dae-Young; Lee, Jung-Hee; Lee, Je-Hwan; Lee, Kyoo-Hyung; Lee, Dae Ho; Suh, Cheolwon; Kim, Sung-Han

    2011-05-01

    There are limited data on the incidence of subsequent bloodstream infection (BSI) and the effect of systemic antibiotics in patients who had positive catheter-drawn blood cultures (CBC) and negative peripheral blood cultures (PBC). We retrospectively reviewed all paired blood cultures from patients with Hickman catheter in the hematology-oncology ward between January 1997 and December 2008. There were 112 episodes with positive CBC and negative PBC. Nine episodes (8.0%; 95% CI, 3.0-13.1%) led to subsequent BSI within 28 days. Subsequent BSI developed in 6 of 31 episodes (19%) where empiric antibiotics were inappropriate but in 3 of 81 episodes (4%) where empiric antibiotics were appropriate (P = 0.01). Subsequent candidemia (50%, 2 of 4) was more common than subsequent bacteremia (6%, 7 of 108) (P = 0.03). In conclusion, for patients with positive CBC and negative PBC, the overall incidence of subsequent BSI was 8.0%, and inappropriate empiric antibiotics was associated with subsequent BSI.

  19. Prediction of systolic blood pressure using peripheral pulse palpation in cats.

    PubMed

    Reineke, Erica L; Rees, Colleen; Drobatz, Kenneth J

    2016-01-01

    To evaluate the ability of peripheral pulse palpation to predict systolic blood pressure (SBP) in cats presenting as emergencies. Prospective observational study performed over an 8-month period. University veterinary teaching hospital. One hundred two cats presenting to the emergency service. Eligibility for inclusion in the study included a physical examination and a SBP via Doppler technique performed prior to treatment. None. Femoral and metatarsal pulses were digitally palpated and the quality of the pulses was assessed as either strong, moderate, poor, or absent. A concurrent SBP was also recorded. The median SBP for all cats was 92.5 mm Hg (range, 30-240 mm Hg). Femoral pulse quality was found to strongly correlate with the admission SBP (P < 0.001, rho = 0.6755). The median SBP for each femoral pulse quality category (strong, moderate, poor, or absent) was significantly different (P < 0.05). For metatarsal pulses, the median SBP for cats with either absent or strong pulses was significantly different (P < 0.001). Cats with absent metatarsal and femoral pulses had a median SBP of 30 mm Hg (range, 30-105 mm Hg), whereas cats with strong metatarsal pulses had a median SBP of 135 mm Hg (range, 58-210 mm Hg). Absent metatarsal pulses correctly identified cats with a blood pressure of 75 mm Hg or less 84% the time (area under the curve: 0.89, confidence interval 0.81, 0.97). In cats, peripheral pulse quality assessment by emergency room veterinarians correlates with SBP. With progressive decreases in blood pressure, metatarsal pulses will disappear and it is only with severe hypotension that femoral pulses are absent. An assessment of both dorsal metatarsal pulse and femoral pulse quality during triage may be useful in identifying abnormalities in blood pressure. © Veterinary Emergency and Critical Care Society 2015.

  20. Apoptotic effects of tamoxifen on leukocytes from horse peripheral blood and bronchoalveolar lavage fluid.

    PubMed

    Sarmiento, J; Perez, B; Morales, N; Henriquez, C; Vidal, L; Folch, H; Galecio, J S; Morán, G

    2013-12-01

    A reduction in inflammatory cell apoptosis is an important concept in the maintenance of inflammation and a potential target for the resolution of inflammation in many inflammatory diseases. Dysregulation of apoptosis has been implicated in a range of diseases, including tumors, neurodegenerative disorders and autoimmunity, and may also be implicated in allergic asthma. In horses, recurrent airway obstruction (RAO) is an asthma-like condition that is characterized increased survival neutrophil bronchial. Tamoxifen is a synthetic, non-steroidal, anti-estrogen agent that is widely used for treating all stages of breast cancer and has been approved for the prevention of breast cancer in high-risk women. The observed efficacy of tamoxifen has been attributed to both growth arrest and the induction of apoptosis. Therefore, the aim of our study was to evaluate the ability of tamoxifen to induce apoptosis in vitro in granulocytic cells from peripheral blood and in mononuclear cells from bronchoalveolar lavage fluid (BALF) in horses. Flow cytometry using commercial AnnexinV-FITC and propidium iodide was used to quantify early and late apoptotic leukocytes, respectively. The results showed a significant increase in early apoptosis in peripheral blood and bronchial granulocytic cells treated with tamoxifen. The rate of early apoptosis of mononuclear cells from blood and BALF when incubated with tamoxifen was significantly lower compared with granulocytic cells. We did not observe a direct effect of tamoxifen on late apoptosis in any of the in vitro assays in the cell types used here. These results indicate that the apoptotic mechanisms under these experimental conditions would affect only blood and BALF granulocytic cells, particularly in early apoptosis. Finally, further in vitro and in vivo studies are needed to better understand apoptotic mechanisms because tamoxifen could be used to treat chronic, inflammatory pathologies associated with granulocytes and allergic

  1. Characterization of endothelial colony-forming cells from peripheral blood samples of adult horses.

    PubMed

    Salter, Margaret M; Seeto, Wen J; DeWitt, Blake B; Hashimi, Sarah A; Schwartz, Dean D; Lipke, Elizabeth A; Wooldridge, Anne A

    2015-02-01

    To isolate and characterize endothelial colony-forming cells (ECFCs; a subtype of endothelial progenitor cells) from peripheral blood samples of horses. Jugular venous blood samples from 24 adult horses. Blood samples were cultured in endothelial cell growth medium. Isolated ECFCs were characterized by use of functional assays of fluorescence-labeled acetylated low-density lipoprotein (DiI-Ac-LDL) uptake and vascular tubule formation in vitro. Expression of endothelial (CD34, CD105, vascular endothelial growth factor receptor 2, and von Willebrand factor) and hematopoietic (CD14) cell markers was assessed through indirect immunofluorescence assay and flow cytometry. The number of passages before senescence was determined through serial evaluation of DiI-Ac-LDL uptake, vascular tubule formation, and cell doubling rates. Samples from 3 horses produced colonies at 12 ± 2.5 days with characteristic endothelial single layer cobblestone morphology and substantial outgrowth on expansion. Equine ECFCs formed vascular tubules in vitro and had uptake of DiI-Ac-LDL (74.9 ± 14.7% positive cells). Tubule formation and DiI-Ac-LDL uptake diminished by passage 5. Equine ECFCs tested positive for von Willebrand factor, vascular endothelial growth factor receptor 2, CD34, and CD105 with an immunofluorescence assay and for CD14 and CD105 via flow cytometry. ECFCs can be isolated from peripheral blood of horses and have characteristics similar to those described for other species. These cells may have potential therapeutic use in equine diseases associated with ischemia or delayed vascularization.

  2. Peripheral blood and bone marrow changes in patients with acquired immunodeficiency syndrome.

    PubMed

    Adediran, I A; Durosinmi, M A

    2006-12-01

    There is dearth of information on the haematologic complications of HIV/AIDS in this country. The aim of this work was to evaluate the importance of peripheral blood and bone marrow changes in a population of adult Nigerians managed for symptomatic HIV infection at the OAUTHC, Ile-Ife, between 1995 and 2001. The study was prospective. Peripheral blood cells counts (haemetocrit value, total and differential leucocyte counts, and platelet counts) and bone marrow cytology of serologically confirmed HIV/AIDS patients seen within the study period were studied. The associated opportunistic disorder (s) was noted for each patient. Individuals with conditions that are ordinarily associated with haematologic disorders (e.g. cancer and inherited haemoglobinopathies) and patients diagnosed in pregnancy were not included. Significant levels of differences in mean values of blood cells within groups were determined by student's t-test. Seventy-two patients were recorded, out of which 49 (68%) were evaluable. There were 32 (65%) males and 17 (35%) females, all aged between 21 and 51 (median = 36) years. None of the patients had conventional antiretroviral therapy. Lymphopenia (lymphocytes < 2 x 10(9)/l) was seen in 64.4% of the patients, 50% and over 40% of the patients had moderate-severe anaemia and neutropenia, respectively. Blood cells values were not significantly different between patients with mild disease and those with moderate-severe diseases. The most characteristic marrow abnormality was the abundance of naked nuclei of megakarycytes in 20 (60.1%) of the patients. Dysplatic changes were evident in 15 (45.5%) of the bone marrow specimens studied. Such changes are characterised by dysgranulopoiesis, Pelger-Huet anomaly in some of the mature granulocytes, vacuolation of some erythroid and myeloid cells, unilobular micromegakaryocytes and megaloblastic erythroid precursors (15.1% of the marrow).

  3. Radionuclide assessment of peripheral hemodynamics: a new technique for measurement of forearm blood volume and flow

    SciTech Connect

    Todo, Y.; Tanimoto, M.; Yamamoto, T.; Iwasaki, T.

    1986-02-01

    A new peripheral hemodynamic measurement system using /sup 99m/Tc-labeled red blood cells has been developed. This method was carried out on 22 normal subjects, 29 with coronary artery disease, and two with dilated cardiomyopathy. Peripheral hemodynamic indices obtained from this method included forearm blood volume (FBV), venous capacity (FVC), venous capacity index (VCI), blood flow (FBF), and vascular resistance (FVR), and were compared with the central hemodynamic parameters of left ventricular filling pressure (LVFP), cardiac output (CO), and total systemic vascular resistance (TSVR) obtained with an invasive technique. The normal values were FBV 8.54 +/- 2.04 ml/100 ml; FVC 4.54 +/- 1.23 ml/100 ml; VCI 65.5 +/- 3.8%; FBF 4.26 +/- 0.56 ml/100 ml/min; and FVR 20.9 +/- 4.4 mmHg/ml/100 ml/min. These values were in good agreement with the values reported using conventional plethysmography. The 16 patients with congestive heart failure (NYHA Class II or III) showed significantly lower FBV, FVC, and FBF values and significantly higher VCI and FVR values than the healthy subjects. Capacitance vessel parameters (FBV, FVC, and VCI) and LVFP, FBF and CO, and FVR and TSVR each showed significant correlation; reproducibility was also good. The advantages of this method are (a) the detector does not come in contact with the region being measured; (b) it is possible to ascertain the absolute quantity of blood in the tissue; (c) extravasation of the plasma component can be ignored; and (d) data processing is simple.

  4. Peripheral Blood MCEMP1 Gene Expression as a Biomarker for Stroke Prognosis.

    PubMed

    Raman, Kripa; O'Donnell, Martin J; Czlonkowska, Anna; Duarte, Yan Carlos; Lopez-Jaramillo, Patricio; Peñaherrera, Ernesto; Sharma, Mike; Shoamanesh, Ashkan; Skowronska, Marta; Yusuf, Salim; Paré, Guillaume

    2016-03-01

    A limitation when making early decisions on stroke management is the lack of rapid diagnostic and prognostic testing. Our study sought to identify peripheral blood RNA biomarkers associated with stroke. The secondary aims were to assess the discriminative capacity of RNA biomarkers for primary stroke type and stroke prognosis at 1-month. Whole-blood gene expression profiling was conducted on the discovery cohort: 129 first-time stroke cases that had blood sampling within 5 days of symptom onset and 170 control participants with no history of stroke. Through multiple regression analysis, we determined that expression of the gene MCEMP1 had the strongest association with stroke of 11 181 genes tested. MCEMP1 increased by 2.4-fold in stroke when compared with controls (95% confidence interval, 2.0-2.8; P=8.2×10(-22)). In addition, expression was elevated in intracerebral hemorrhage when compared with ischemic stroke cases (P=3.9×10(-4)). MCEMP1 was also highest soon after symptom onset and had no association with stroke risk factors. Furthermore, MCEMP1 expression independently improved discrimination of 1-month outcome. Indeed, discrimination models for disability and mortality that included MCEMP1 expression, baseline modified Rankin Scale score, and primary stroke type improved discrimination when compared with a model without MCEMP1 (disability Net Reclassification Index, 0.76; P=3.0×10(-6) and mortality Net Reclassification Index, 1.3; P=1.1×10(-9)). Significant associations with MCEMP1 were confirmed in an independent validation cohort of 28 stroke cases and 34 controls. This study demonstrates that peripheral blood expression of MCEMP1 may have utility for stroke diagnosis and as a prognostic biomarker of stroke outcome at 1-month. © 2016 American Heart Association, Inc.

  5. [Peripheral blood circulation in the skin and the regulatory mechanisms in the course of primary transmural myocardial infarction].

    PubMed

    Khalepo, O V; Molotkov, O V; Eshkina, S L

    2009-01-01

    Laser Doppler flowmetry was used to study the indicators characterizing the peripheral blood circulation in the skin, regulatory mechanisms, and the compensatory capacities of the microcirculatory bed in 32 patients aged 45-60 years in the course of primary transmural myocardial infarction during exercise tests. Significant disturbances of the mechanism responsible for regulating the peripheral blood circulation system and chiefly its active components were detected in the presence of adequate blood filling of microvessels. There was a drastic decrease in the reserves of skin microvascular endothelial activity during ionophoresis of sodium nitroprusside and acetylcholine, the maximum degree of disturbances being observed on day 10 of myocardial infarction development.

  6. Influence of blood flow occlusion on the development of peripheral and central fatigue during small muscle mass handgrip exercise.

    PubMed

    Broxterman, R M; Craig, J C; Smith, J R; Wilcox, S L; Jia, C; Warren, S; Barstow, T J

    2015-09-01

    Critical power represents an important threshold for neuromuscular fatigue development and may, therefore, dictate intensities for which exercise tolerance is determined by the magnitude of fatigue accrued. Peripheral fatigue appears to be constant across O2 delivery conditions for large muscle mass exercise, but this consistency is equivocal for smaller muscle mass exercise. We sought to determine the influence of blood flow occlusion during handgrip exercise on neuromuscular fatigue development and to examine the relationship between neuromuscular fatigue development and W '. Blood flow occlusion influenced the development of both peripheral and central fatigue, thus providing further evidence that the magnitude of peripheral fatigue is not constant across O2 delivery conditions for small muscle mass exercise. W ' appears to be related to the magnitude of fatigue accrued during exercise, which may explain the reported consistency of intramuscular metabolic perturbations and work performed for severe-intensity exercise. The influence of the muscle metabolic milieu on peripheral and central fatigue is currently unclear. Moreover, the relationships between peripheral and central fatigue and the curvature constant (W ') have not been investigated. Six men (age: 25 ± 4 years, body mass: 82 ± 10 kg, height: 179 ± 4 cm) completed four constant power handgrip tests to exhaustion under conditions of control exercise (Con), blood flow occlusion exercise (Occ), Con with 5 min post-exercise blood flow occlusion (Con + Occ), and Occ with 5 min post-exercise blood flow occlusion (Occ + Occ). Neuromuscular fatigue measurements and W ' were obtained for each subject. Each trial resulted in significant peripheral and central fatigue. Significantly greater peripheral (79.7 ± 5.1% vs. 22.7 ± 6.0%) and central (42.6 ± 3.9% vs. 4.9 ± 2.0%) fatigue occurred for Occ than for Con. In addition, significantly greater peripheral (83.0 ± 4.2% vs. 69.0 ± 6.2%) and central

  7. Blood sampling through peripheral venous catheters is reliable for selected basic analytes in children.

    PubMed

    Berger-Achituv, Sivan; Budde-Schwartzman, Britta; Ellis, Martin H; Shenkman, Ze'ev; Erez, Ilan

    2010-07-01

    The goal was to determine the interchangeability of peripheral venous catheter (PVC) and venipuncture blood sampling (BS). Paired blood samples from hospitalized children were obtained through venipuncture and from existing PVCs, following discard of 2 mL of blood. Comparisons of 9 complete blood count indices (white and red blood cell counts, hemoglobin and hematocrit levels, mean corpuscular volume, mean corpuscular hemoglobin level, red blood cell distribution width, platelet count, and mean platelet volume) and 5 basic chemical analysis indices (sodium, potassium, glucose, chloride, and urea levels) were performed, and hemolysis was documented. Irrespective of gauge, blood samples were obtained successfully from 40 (85.1%) of 47 PVCs, with no abnormal hemolysis. BS through venipuncture took longer than BS from PVCs (175.8 +/- 229.6 vs 104.5 +/- 53.4 seconds; P = .053) and was associated with significantly more distress/crying (73.1% vs 0%; P < .001). There were no significant differences between venipuncture and PVC samples (paired t test). Twenty-one (6%) of 348 pairs analyzed with the Clinical Laboratory Improvement Amendment standards fell outside the range of acceptable variance (8 of 21 aberrations were attributed to glucose measurements). Bland-Altman analysis indicated that, with the exclusion of glucose measurements, BS from PVCs is reliable, with 29 (6.5%) of 448 pairs exceeding the limits of agreement. Of those, 9 cases were clinically significant, but none would have altered clinical management. PVC sampling was shown to be a pain-reducing method that can be used for children for selected basic analytes. The findings for glucose were unreliable.

  8. Molecular Classifiers for Acute Kidney Transplant Rejection in Peripheral Blood by Whole Genome Gene Expression Profiling

    PubMed Central

    Kurian, S. M.; Williams, A. N.; Gelbart, T.; Campbell, D.; Mondala, T. S.; Head, S. R.; Horvath, S.; Gaber, L.; Thompson, R.; Whisenant, T.; Lin, W.; Langfelder, P.; Robison, E. H.; Schaffer, R. L.; Fisher, J. S.; Friedewald, J.; Flechner, S. M.; Chan, L. K.; Wiseman, A. C.; Shidban, H.; Mendez, R.; Heilman, R.; Abecassis, M. M.; Marsh, C. L.; Salomon, D. R.

    2015-01-01

    There are no minimally invasive diagnostic metrics for acute kidney transplant rejection (AR), especially in the setting of the common confounding diagnosis, acute dysfunction with no rejection (ADNR). Thus, though kidney transplant biopsies remain the gold standard, they are invasive, have substantial risks, sampling error issues and significant costs and are not suitable for serial monitoring. Global gene expression profiles of 148 peripheral blood samples from transplant patients with excellent function and normal histology (TX; n = 46), AR (n = 63) and ADNR (n = 39), from two independent cohorts were analyzed with DNA microarrays. We applied a new normalization tool, frozen robust multi-array analysis, particularly suitable for clinical diagnostics, multiple prediction tools to discover, refine and validate robust molecular classifiers and we tested a novel one-by-one analysis strategy to model the real clinical application of this test. Multiple three-way classifier tools identified 200 highest value probesets with sensitivity, specificity, positive predictive value, negative predictive value and area under the curve for the validation cohort ranging from 82% to 100%, 76% to 95%, 76% to 95%, 79% to 100%, 84% to 100% and 0.817 to 0.968, respectively. We conclude that peripheral blood gene expression profiling can be used as a minimally invasive tool to accurately reveal TX, AR and ADNR in the setting of acute kidney transplant dysfunction. PMID:24725967

  9. Erythrocyte dysplasia in peripheral blood smears from 5 thrombocytopenic dogs treated with vincristine sulfate.

    PubMed

    Collicutt, Nancy B; Garner, Bridget

    2013-12-01

    Secondary dyserythropoiesis has been associated with vincristine administration in dogs. Evaluation of bone marrow aspirates for the presence of morphologic abnormalities in the erythroid lineage aids in the diagnosis. However, morphologic features of circulating erythroid precursors in these cases have not been described previously. The purpose of this report was to describe the cytologic features of dyserythropoiesis in peripheral blood and also bone marrow smears in a case series of dogs with immune-mediated thrombocytopenia (IMT) treated with vincristine sulfate. Nineteen dogs receiving vincristine for treatment of IMT were identified by retrospectively searching a computerized medical record system. There were 5 dogs that had dysplastic erythroid precursors in peripheral blood smears within 7 days of vincristine treatment. Two of those 5 dogs also had evidence for erythrodysplasia in modified Wright's-stained bone marrow smears obtained postvincristine administration. Morphologic changes included bizarre or inappropriate mitotic figures, abnormal nuclear configurations (fragmentation, elongation, indentation, and binucleation), atypical nuclear remnants (Howell-Jolly bodies), or nuclear and cytoplasmic asynchrony within the erythroid precursors. A brief review of the literature with discussion of the etiologies for dyserythropoiesis is provided. The dyserythropoiesis was clinically insignificant in all 5 cases and resolved. However, pathologists and clinicians should be aware of these potential findings to prevent misdiagnosis of other conditions.

  10. Huntington's disease biomarker progression profile identified by transcriptome sequencing in peripheral blood.

    PubMed

    Mastrokolias, Anastasios; Ariyurek, Yavuz; Goeman, Jelle J; van Duijn, Erik; Roos, Raymund A C; van der Mast, Roos C; van Ommen, GertJan B; den Dunnen, Johan T; 't Hoen, Peter A C; van Roon-Mom, Willeke M C

    2015-10-01

    With several therapeutic approaches in development for Huntington's disease, there is a need for easily accessible biomarkers to monitor disease progression and therapy response. We performed next-generation sequencing-based transcriptome analysis of total RNA from peripheral blood of 91 mutation carriers (27 presymptomatic and, 64 symptomatic) and 33 controls. Transcriptome analysis by DeepSAGE identified 167 genes significantly associated with clinical total motor score in Huntington's disease patients. Relative to previous studies, this yielded novel genes and confirmed previously identified genes, such as H2AFY, an overlap in results that has proven difficult in the past. Pathway analysis showed enrichment of genes of the immune system and target genes of miRNAs, which are downregulated in Huntington's disease models. Using a highly parallelized microfluidics array chip (Fluidigm), we validated 12 of the top 20 significant genes in our discovery cohort and 7 in a second independent cohort. The five genes (PROK2, ZNF238, AQP9, CYSTM1 and ANXA3) that were validated independently in both cohorts present a candidate biomarker panel for stage determination and therapeutic readout in Huntington's disease. Finally we suggest a first empiric formula predicting total motor score from the expression levels of our biomarker panel. Our data support the view that peripheral blood is a useful source to identify biomarkers for Huntington's disease and monitor disease progression in future clinical trials.

  11. A gene expression signature for recent onset rheumatoid arthritis in peripheral blood mononuclear cells

    PubMed Central

    Olsen, N; Sokka, T; Seehorn, C; Kraft, B; Maas, K; Moore, J; Aune, T

    2004-01-01

    Background: In previous studies the presence of a distinct gene expression pattern has been shown in peripheral blood cells from patients with autoimmune disease. Objective: To determine whether other specific signatures might be used to identify subsets of these autoimmune diseases and whether gene expression patterns in early disease might identify pathogenetic factors. Methods: Peripheral blood mononuclear cells were acquired from patients with rheumatoid arthritis (RA) and analysed by microarrays containing over 4300 named human genes. Patients with RA for <2 years were compared with subjects with longstanding RA (average duration 10 years) and with patients with other immune or autoimmune diagnoses. Results: Cluster analyses permitted separation of the patients with early RA (ERA) from those with longstanding disease. Comparison with other patient groups suggested that the ERA signature showed some overlap with that seen in the normal immune response to viral antigen as well as with a subset of patients with systemic lupus erythematosus. Conclusions: The ERA signature may reflect, in part, a response to an unknown infectious agent. Furthermore, shared features with some lupus patients suggest that common aetiological factors and pathogenetic pathways may be involved in these two autoimmune disorders. PMID:15479887

  12. Regulation of Exacerbated Immune Responses in Human Peripheral Blood Cells by Hydrolysed Egg White Proteins

    PubMed Central

    Lozano-Ojalvo, Daniel; Molina, Elena; López-Fandiño, Rosina

    2016-01-01

    The anti-allergic potential of egg white protein hydrolysates (from ovalbumin, lysozyme and ovomucoid) was evaluated as their ability to hinder cytokine and IgE production by Th2-skewed human peripheral blood mononuclear cells (PBMCs), as well as the release of pro-inflammatory factors and generation of reactive oxygen species from Th1-stimulated peripheral blood leukocytes (PBLs). The binding to IgE of egg allergic patients was determined and the peptides present in the hydrolysates were identified. The hydrolysates with alcalase down-regulated the production of Th2-biased cytokines and the secretion of IgE to the culture media of Th2-skewed PBMCs, and they significantly neutralized oxidative stress in PBLs. The hydrolysates of ovalbumin and ovomucoid with pepsin helped to re-establish the Th1/Th2 balance in Th2-biased PBMCs, while they also inhibited the release of pro-inflammatory mediators and reduced oxidative stress in PBLs treated with inflammatory stimuli. The hydrolysates with alcalase, in addition to equilibrating Th2 differentiation, exhibited a low IgE-binding. Therefore, they would elicit mild allergic reactions while retaining T cell-stimulating abilities, which might correlate with an anti-allergic benefit. PMID:27007699

  13. Pubertal development in healthy children is mirrored by DNA methylation patterns in peripheral blood

    PubMed Central

    Almstrup, Kristian; Lindhardt Johansen, Marie; Busch, Alexander S.; Hagen, Casper P.; Nielsen, John E.; Petersen, Jørgen Holm; Juul, Anders

    2016-01-01

    Puberty marks numerous physiological processes which are initiated by central activation of the hypothalamic–pituitary–gonadal axis, followed by development of secondary sexual characteristics. To a large extent, pubertal timing is heritable, but current knowledge of genetic polymorphisms only explains few months in the large inter-individual variation in the timing of puberty. We have analysed longitudinal genome-wide changes in DNA methylation in peripheral blood samples (n = 102) obtained from 51 healthy children before and after pubertal onset. We show that changes in single methylation sites are tightly associated with physiological pubertal transition and altered reproductive hormone levels. These methylation sites cluster in and around genes enriched for biological functions related to pubertal development. Importantly, we identified that methylation of the genomic region containing the promoter of TRIP6 was co-ordinately regulated as a function of pubertal development. In accordance, immunohistochemistry identified TRIP6 in adult, but not pre-pubertal, testicular Leydig cells and circulating TRIP6 levels doubled during puberty. Using elastic net prediction models, methylation patterns predicted pubertal development more accurately than chronological age. We demonstrate for the first time that pubertal attainment of secondary sexual characteristics is mirrored by changes in DNA methylation patterns in peripheral blood. Thus, modulations of the epigenome seem involved in regulation of the individual pubertal timing. PMID:27349168

  14. Peripheral Blood Mononuclear Cells as a Laboratory to Study Dementia in the Elderly

    PubMed Central

    Arosio, Beatrice; D'Addario, Claudio; Gussago, Cristina; Casati, Martina; Tedone, Enzo; Ferri, Evelyn; Nicolini, Paola; Rossi, Paolo D.; Maccarrone, Mauro; Mari, Daniela

    2014-01-01

    The steady and dramatic increase in the incidence of Alzheimer's disease (AD) and the lack of effective treatments have stimulated the search for strategies to prevent or delay its onset and/or progression. Since the diagnosis of dementia requires a number of established features that are present when the disease is fully developed, but not always in the early stages, the need for a biological marker has proven to be urgent, in terms of both diagnosis and monitoring of AD. AD has been shown to affect peripheral blood mononuclear cells (PBMCs) that are a critical component of the immune system which provide defence against infection. Although studies are continuously supplying additional data that emphasize the central role of inflammation in AD, PBMCs have not been sufficiently investigated in this context. Delineating biochemical alterations in AD blood constituents may prove valuable in identifying accessible footprints that reflect degenerative processes within the Central Nervous System (CNS). In this review, we address the role of biomarkers in AD with a focus on the notion that PBMCs may serve as a peripheral laboratory to find molecular signatures that could aid in differential diagnosis with other forms of dementia and in monitoring of disease progression. PMID:24877062

  15. Negative immunomagnetic selection of T cells from peripheral blood of presentation AML specimens.

    PubMed

    Le Dieu, Rifca; Taussig, David; Lister, T Andrew; Gribben, John G

    2009-08-31

    To date, studies on T cells in acute myeloid leukemia (AML) have been limited to flow cytometric analysis of whole peripheral blood mononuclear cell (PBMC) specimens or functional work looking at the impact of AML myeloblasts on normal or remission T cells. This lack of information on T cells at the time of presentation with disease is due in part to the difficulty in isolating sufficiently pure T cells from these specimens for further study. Negative immunomagnetic selection has been the method of choice for isolating immune cells for functional studies due to concerns that binding antibodies to the cell surface may induce cellular activation, block ligand-receptor interactions or result in immune clearance. In order specifically to study T cells in presentation AML specimens, we set out to develop a method of isolating highly pure CD4 and CD8 T cells by negative selection from the peripheral blood (PB) of newly diagnosed AML patients. This technique, unlike T cell selection from PB from normal individuals or from patients with chronic lymphocytic leukaemia, was extremely problematic due to properties of the leukaemic myeloblasts. A successful method was eventually optimized requiring the use of a custom antibody cocktail consisting of CD33, CD34, CD123, CD11c and CD36, to deplete myeloblasts.

  16. Does the DASH diet lower blood pressure by altering peripheral vascular function?

    PubMed

    Hodson, L; Harnden, K E; Roberts, R; Dennis, A L; Frayn, K N

    2010-05-01

    We tested whether lowering of blood pressure (BP) on the dietary approaches to stop hypertension (DASH) diet was associated with changes in peripheral vascular function: endothelial function, assessed by flow-mediated vasodilatation (FMD) of the brachial artery, and subcutaneous adipose tissue blood flow (ATBF). We also assessed effects on heart rate variability (HRV) as a measure of autonomic control of the heart. We allocated 27 men and women to DASH diet and control groups. We measured FMD, ATBF and HRV on fasting and after ingestion of 75 g glucose, before and after 30 days on dietary intervention, aiming for weight maintenance. The control group did not change their diet. The DASH-diet group complied with the diet as shown by significant reductions in systolic (P<0.001) and diastolic (P=0.005) BP, and in plasma C-reactive protein (P<0.01), LDL-cholesterol (P<0.01) and apolipoprotein B (P=0.001), a novel finding. Body weight changed by <1 kg. There were no changes in the control group. We found no changes in FMD, or in ATBF, in the DASH-diet group, although heart rate fell (P<0.05). Glucose and insulin concentrations did not change. In this small-scale study, the DASH diet lowered BP independently of peripheral mechanisms.

  17. Coproporphyrinogen oxidase activity and porphyrin concentrations in peripheral red blood cells in hereditary sideroblastic anaemia.

    PubMed

    Pasanen, A V; Eklöf, M; Tenhunen, R

    1985-03-01

    The activity of coproporphyrinogen oxidase and the concentrations of coproporphyrin and protoporphyrin (measured by HPLC) in peripheral red blood cells were established in 2 families with different types of hereditary sideroblastic anaemia. 2 males and 4 females were members of a family with an X-chromosome-linked and pyridoxine-responsive HSA, and 3 females were members of another family where the mode of inheritance is not clear and where pyridoxine did not produce a haematological response. Coproporphyrinogen oxidase activity was normal in 8 of 9 patients and slightly decreased only in 1 patient. All patients had normal red cell coproporphyrin concentrations, but red cell protoporphyrin concentration was decreased in 4 patients. These findings indicate that in vivo haem synthesis was not impaired at the step of coproporphyrinogen oxidase, hence enzymatic defects in earlier steps of haem synthesis are more evident. Earlier suggestions of impaired haem synthesis at this level, based on observed increased concentrations of coproporphyrin in peripheral red blood cells might be explained by the use of unspecific methods.

  18. Persistent Mosaicism for 12p Duplication/Triplication Chromosome Structural Abnormality in Peripheral Blood

    PubMed Central

    Shackelford, Amy L.; Conlin, Laura K.; Spinner, Nancy B.; Wenger, Sharon L.

    2013-01-01

    We present a rare case of mosaicism for a structural abnormality of chromosome 12 in a patient with phenotypic features of Pallister-Killian syndrome. A six-month-old child with dysmorphic features, exotropia, hypotonia, and developmental delay was mosaic for both a normal karyotype and a cell line with 12p duplication/triplication in 25 percent of metaphase cells. Utilization of fluorescence in situ hybridization (FISH) identified three copies of probes from the end of the short arm of chromosome 12 (TEL(12p13) locus and the subtelomere (12p terminal)) on the structurally abnormal chromosome 12. Genome-wide SNP array analysis revealed that the regions of duplication and triplication were of maternal origin. The abnormal cell line in our patient was present at 25 percent at six months and 19 months of age in both metaphase and interphase cells from peripheral blood, where typically the isochromosome 12p is absent in the newborn. This may suggest that the gene(s) resulting in a growth disadvantage of abnormal cells in peripheral blood of patients with tetrasomy 12p may not have the same influence when present in only three copies. PMID:24151566

  19. Characterization of peripheral blood human immunodeficiency virus isolates from Hispanic women with cognitive impairment.

    PubMed

    Nieves, Dianedis M Toro; Plaud, Marinés; Wojna, Valerie; Skolasky, Richard; Meléndez, Loyda M

    2007-08-01

    Human immunodeficiency virus type 1 (HIV-1) tropism plays an important role in HIV-associated dementia. In this study, aimed at determining if the tropism and coreceptor usage of circulating viruses correlates with cognitive function, the authors isolated and characterized HIV from the peripheral blood of 21 Hispanic women using antiretroviral therapy. Macrophage tropism was determined by inoculation of HIV isolates onto monocyte-derived macrophages and lymphocyte cultures. To define coreceptor usage, the HIV isolates were inoculated onto the U87.CD4 glioma cell lines with specific CCR5 and CXCR4 coreceptors. HIV isolates from cognitively impaired patients showed higher levels of replication in mitogen-stimulated peripheral blood mononuclear cells than did isolates from patients with normal cognition (P < .05). The viral growth of HIV primary isolates in macrophages and lymphocytes did not differ between patients with and those without cognitive impairment. However, isolates from the cognitively impaired women preferentially used the X4 coreceptor (P < .05). These phenotypic studies suggest that cognitively impaired HIV-infected women receiving treatment may have a more highly replicating and more pathogenic X4 virus in the circulation that could contribute to their neuropathogenesis.

  20. Modifications of the endosomal compartment in peripheral blood mononuclear cells and fibroblasts from Alzheimer's disease patients

    PubMed Central

    Corlier, F; Rivals, I; Lagarde, J; Hamelin, L; Corne, H; Dauphinot, L; Ando, K; Cossec, J-C; Fontaine, G; Dorothée, G; Malaplate-Armand, C; Olivier, J-L; Dubois, B; Bottlaender, M; Duyckaerts, C; Sarazin, M; Potier, M-C; Alnajjar-Carpentier, Dr Amer; Logak, Dr Michel; Leder, Dr Sara; Marchal, Dr Dominique; Pitti-Ferandi, Dr Hélène; Brugeilles, Dr Hélene; Roualdes, Dr Brigitte; Michon, Dr Agnes

    2015-01-01

    Identification of blood-based biomarkers of Alzheimer's disease (AD) remains a challenge. Neuropathological studies have identified enlarged endosomes in post-mortem brains as the earliest cellular change associated to AD. Here the presence of enlarged endosomes was investigated in peripheral blood mononuclear cells from 48 biologically defined AD patients (25 with mild cognitive impairment and 23 with dementia (AD-D)), and 23 age-matched healthy controls using immunocytochemistry and confocal microscopy. The volume and number of endosomes were not significantly different between AD and controls. However, the percentage of cells containing enlarged endosomes was significantly higher in the AD-D group as compared with controls. Furthermore, endosomal volumes significantly correlated to [C11]PiB cortical index measured by positron emission tomography in the AD group, independently of the APOE genotype, but not to the levels of amyloid-beta, tau and phosphorylated tau measured in the cerebrospinal fluid. Importantly, we confirmed the presence of enlarged endosomes in fibroblasts from six unrelated AD-D patients as compared with five cognitively normal controls. This study is the first, to our knowledge, to report morphological alterations of the endosomal compartment in peripheral cells from AD patients correlated to amyloid load that will now be evaluated as a possible biomarker. PMID:26151923

  1. Collection of peripheral progenitor cells: a comparison between Amicus and Cobe-Spectra blood cell separators.

    PubMed

    Adorno, Gaspare; Del Proposto, Gianpaolo; Palombi, Francesca; Bruno, Antonio; Ballatore, Giovanna; Postorino, Massimiliano; Tendas, Andrea; Del Poeta, Giovanni; Isacchi, Giancarlo; Amadori, Sergio

    2004-04-01

    The authors compared the efficiency of two different blood cell separators (Amicus and Cobe-Spectra) in collecting peripheral blood progenitor cells for autologous or homologous transplantation. A total number of 129 procedures were performed, 36 with Spectra, 93 with Amicus. There was no difference between Spectra and Amicus efficiencies for CD34+ cell collection (46.685% vs 46.235%; p=n.s) but the platelet efficiencies were 17.31% and 12.54% respectively (p=0.04) and, if autologous and allogeneic collections were considered separately, a marked difference resulted in allogeneic platelet efficiency between 6 Spectra and 23 Amicus procedures (26.83% vs 8.68%, p=0.0004). The authors were able to demonstrate that in 70 Amicus autologous collections there was a different platelet efficiency, if peripheral count was considered: 12 procedures performed with a platelet count > 100 x 10(9)/l had a very low efficiency (6.86%), but this value increased if platelet count lowered (13.02% if between 100 and 50 x 10(9)/l, 22.63% if between 50 and 0 x 10(9)/l, 23 and 35 procedures respectively). The study is preliminary and the number of collections is little, but the overall data suggest that Spectra (AutoPBSC, V 6.0) and Amicus separators have the same efficiency for collecting CD34+ cells while Amicus procedures have a very low platelet contamination, especially with donors.

  2. Effect of alcohol consumption on peripheral blood Alu methylation in Korean men.

    PubMed

    Kim, Dong-Sun; Kim, Young Hun; Lee, Won Kee; Na, Yeon Kyung; Hong, Hae Sook

    2016-01-01

    Alcohol use disorders (AUD) are defined as alcohol abuse and alcohol dependence, which create a substantial public health problem worldwide. To date, no therapeutic can effectively solve these problems. They are complex diseases characterized by both genetic and environmental factors. DNA methylation can act as a downstream effector of environmental signals and account for multi-factorial nature of the disease. Global DNA methylation of peripheral blood cells has recently been proposed as a potential biomarker for disease risk. Alu elements host one-quarter of CpG dinucelotides in the genome to function as proxies for global DNA methylation. In this study, we evaluated the Alu methylation in the peripheral blood DNA of healthy volunteers and AUD patients using the pyrosequencing technology. The Alu methylation level is significantly higher in AUD compared to healthy controls (23.4 ± 1.6 versus 22.1 ± 1.0, t = 7.83, p < 0.0001). Moreover, significant correlation was found between Alu methylation and alcohol use disorders identification test score (r = 0.250, p < 0.0001), alcohol problem (r = 0.294, p < 0.0001), and life position (r = -0.205, p = 0.0005). Overall, these novel findings indicate that alcohol-related increase in Alu methylation might play a complex role in the etiology and pathogenesis of AUD. Further studies are required to elucidate the mechanisms underlying this relationship.

  3. Peripheral blood lymphocytes: a model for monitoring physiological adaptation to high altitude.

    PubMed

    Mariggiò, Maria A; Falone, Stefano; Morabito, Caterina; Guarnieri, Simone; Mirabilio, Alessandro; Pilla, Raffaele; Bucciarelli, Tonino; Verratti, Vittore; Amicarelli, Fernanda

    2010-01-01

    Depending on the absolute altitude and the duration of exposure, a high altitude environment induces various cellular effects that are strictly related to changes in oxidative balance. In this study, we used in vitro isolated peripheral blood lymphocytes as biosensors to test the effect of hypobaric hypoxia on seven climbers by measuring the functional activity of these cells. Our data revealed that a 21-day exposure to high altitude (5000 m) (1) increased intracellular Ca(2+) concentration, (2) caused a significant decrease in mitochondrial membrane potential, and (3) despite possible transient increases in intracellular levels of reactive oxygen species, did not significantly change the antioxidant and/or oxidative damage-related status in lymphocytes and serum, assessed by measuring Trolox-equivalent antioxidant capacity, glutathione peroxidase activity, vitamin levels, and oxidatively modified proteins and lipids. Overall, these results suggest that high altitude might cause an impairment in adaptive antioxidant responses. This, in turn, could increase the risk of oxidative-stress-induced cellular damage. In addition, this study corroborates the use of peripheral blood lymphocytes as an easily handled model for monitoring adaptive response to environmental challenge.

  4. Defining "poor mobilizer" in pediatric patients who need an autologous peripheral blood progenitor cell transplantation.

    PubMed

    Sevilla, Julián; Guillén, María; Castillo, Ana; Prudencio, Marta; González-Vicent, Marta; Lassaletta, Álvaro; Cormenzana, María; Ramírez, Manuel; Pérez-Martínez, Antonio; Madero, Luis; Díaz-Pérez, Miguel Ángel

    2013-01-01

    The definition of poor mobilizers is not clear in pediatric patients undergoing autologous peripheral blood hematopoietic progenitor cell (HPC) mobilization. Most studies conducted in children define those variables related to the collection of HPC after leukapheresis, but the information regarding exclusively the mobilization process is scarce. In our experience, most children (92.2%) reach the target CD34(+) cell dose for autologous peripheral blood progenitor cell transplantation if CD34(+) cell count was higher than 10/μL. No differences were observed between those with >20 CD34(+) cells/μL and 11-20 CD34(+) cells/μL. In this study, we analyzed the variables that influence CD34(+) cell count; we found that prior use of radiotherapy was the main variable related to poor mobilization. Patients diagnosed with Ewing sarcoma, treated with radiotherapy and mobilized with standard doses of granulocyte colony-stimulating factor (G-CSF) were also at a high risk of mobilization failure. In these patients, we should consider mobilization with high dose G-CSF and be prepared with new mobilization agents to avoid delay on their course of chemotherapy.

  5. Comprehensive assessment of peripheral blood TCRβ repertoire in infectious mononucleosis and chronic active EBV infection patients.

    PubMed

    Liu, Shenglin; Zhang, Qian; Huang, Dongli; Zhang, Wenli; Zhong, Fengluan; Feng, Jia; Chen, Xueru; Meng, Qingxiang; Chen, Xiaofan; Zhang, Wei; Zhang, Hongyu

    2017-04-01

    Epstein-Barr virus (EBV) primary infection is usually asymptomatic, but it sometimes progresses to infectious mononucleosis (IM). Occasionally, some people develop chronic active EBV infection (CAEBV) with underlying immunodeficiency, which belongs to a continuous spectrum of EBV-associated lymphoproliferative disorders (EBV(+) LPD) with heterogeneous clinical presentations and high mortality. It has been well established that T cell-mediated immune response plays a critical role in the disease evolution of EBV infection. Recently, high-throughput sequencing of the hypervariable complementarity-determining region 3 (CDR3) segments of the T cell receptor (T cell receptor β (TCRβ)) has emerged as a sensitive approach to assess the T cell repertoire. In this study, we fully characterized the diversity of peripheral blood TCRβ repertoire in IM (n = 6) and CAEBV patients (n = 5) and EBV-seropositive controls (n = 5). Compared with the healthy EBV-seropositive controls, both IM and CAEBV patients demonstrate a significant decrease in peripheral blood TCRβ repertoire diversity, basically, including narrowed repertoire breadth, highly expanded clones, and skewed CDR3 length distribution. However, there is no significant difference between IM and CAEBV patients. Furthermore, we observed some disease-related preferences in TRBV/TRBJ usage and combinations, as well as lots of T cell clones shared by different groups (unique or overlapped) involved in public T cell responses, which provide more detailed insights into the divergent disease evolution.

  6. Tolerogenic versus Inflammatory Activity of Peripheral Blood Monocytes and Dendritic Cells Subpopulations in Systemic Lupus Erythematosus

    PubMed Central

    Carvalheiro, Tiago; Rodrigues, Ana; Lopes, Ana; Inês, Luís; Velada, Isabel; Ribeiro, Andreia; Martinho, António; Silva, José A. P.; Pais, Maria L.; Paiva, Artur

    2012-01-01

    Abnormalities in monocytes and in peripheral blood dendritic cells (DC) subsets have been reported in systemic lupus erythematosus (SLE). We aim to clarify the tolerogenic or inflammatory role of these cells based on ICOSL or IFN-α and chemokine mRNA expression, respectively, after cell purification. The study included 18 SLE patients with active disease (ASLE), 25 with inactive disease (ISLE), and 30 healthy controls (HG). In purified plasmacytoid DC (pDC) was observed a lower ICOSL mRNA expression in ASLE and an increase in ISLE; similarly, a lower ICOSL mRNA expression in monocytes of ALSE patients was found. However, a higher ICOSL mRNA expression was observed in ASLE compared to HG in myeloid DCs. Interestingly, clinical parameters seem to be related with ICOSL mRNA expression. Regarding the inflammatory activity it was observed in purified monocytes and CD14−/low CD16+ DCs an increase of CCL2, CXCL9, and CXCL10 mRNA expression in ASLE compared to HG. In myeloid DC no differences were observed regarding chemokines, and IFN-α mRNA expression. In pDC, a higher IFN-α mRNA expression was observed in ASLE. Deviations in ICOSL, chemokine, and IFN-α mRNA expression in peripheral blood monocytes and dendritic cells subpopulations in SLE appear to be related to disease activity. PMID:22969819

  7. The Immunomodulatory Effects of Nidus Vespae on Human Peripheral Blood Immune Cells In Vitro

    PubMed Central

    Zhu, Ming; Ling, Yang; Qi, Qiufeng; Zhang, Yaping; Bao, Yanqing; Liu, Yongping

    2015-01-01

    Nidus Vespae has been used in traditional Chinese medicine (TCM) to treat various cancers, but the underlying mechanisms were not yet clarified. This study was to investigate the effect of Nidus Vespae decoction (NVD) on tumor cell viability and immunoregulating functions of human peripheral blood immune cells. The effects on tumor cell viability, peripheral blood mononuclear cell (PBMC) proliferation activity, and the tumor cell phagocytosis of monocytes were evaluated by cell counting kit-8. Tumor-killing activity of cytotoxic T lymphocyte (CTL) was analyzed by 51Cr releasing assay. IgG production of B cells and cytokine (TNF-α and IL-6) secretion of monocytes were determined by ELISA method. Data showed that NVD has no significant inhibiting effects on gastric cancer cells growth. Nevertheless, it could obviously promote PBMC proliferation in a time- and concentration-dependent manner. After treatment with NVD, the CTL cytotoxicity against SGC-7901 was significantly greater than control. The TNF-α and IL-6 secretion of monocytes and the IgG production of B cells also increased remarkably. Furthermore, NVD could significantly promote the phagocytosis of monocytes on tumor cells. These results suggest that NVD appears to have an immunoenhancing effect on immune cells, indicating that Nidus Vespae is worth exploring for immunomodulatory effects in tumor treatment. PMID:26339270

  8. High-dose chemotherapy with peripheral blood stem cell rescue in blastoid natural killer cell lymphoma.

    PubMed

    Mukai, H Y; Kojima, H; Suzukawa, K; Hori, M; Komeno, T; Hasegawa, Y; Ninomiya, H; Mori, N; Nagasawa, T

    1999-02-01

    A 25-year-old man was referred because of skin rash, lymphadenopathy and anemia. Laboratory examinations revealed severe anemia (Hb, 4.8 g/dl) and elevated levels of GOT, GPT, LDH and soluble interleukin-2 receptor. Work-up studies disclosed the involvement of lymphoma cells in lymph nodes, skin, bilateral kidneys and bone marrow. Lymph node biopsy revealed diffuse proliferation of medium- to large-sized lymphoblastic cells. Bone marrow aspiration showed massive infiltration of large blastic cells with no cytoplasmic granules. The lymphoma cells in bone marrow and lymph node showed surface CD3-, cytoplasmic CD3epsilon+, CD4+, CD8-, CD56+, CD57-, CD16- and CD43 (MT-1)+ phenotype. Analyses of T cell receptor beta and gamma genes showed germ line configurations. EBER-1 was not detectable in the lymphoma cells. He was diagnosed as having blastoid natural killer (NK) cell lymphoma. In spite of several courses of combination chemotherapy, the lymphoma was progressive. He was then treated with high-dose chemotherapy and peripheral blood stem cell rescue, achieving remission which has now lasted for more than 12 months. We consider that blastoid NK cell lymphoma is an extremely aggressive subtype of CD56-positive lymphomas, and high-dose chemotherapy with peripheral blood stem cell rescue should be included for the choice of the treatment.

  9. MPL expression on AML blasts predicts peripheral blood neutropenia and thrombocytopenia.

    PubMed

    Rauch, Philipp J; Ellegast, Jana M; Widmer, Corinne C; Fritsch, Kristin; Goede, Jeroen S; Valk, Peter J M; Löwenberg, Bob; Takizawa, Hitoshi; Manz, Markus G

    2016-11-03

    Although the molecular pathways that cause acute myeloid leukemia (AML) are increasingly well understood, the pathogenesis of peripheral blood cytopenia, a major cause of AML mortality, remains obscure. A prevailing assumption states that AML spatially displaces nonleukemic hematopoiesis from the bone marrow. However, examining an initial cohort of 223 AML patients, we found no correlation between bone marrow blast content and cytopenia, questioning the displacement theory. Measuring serum concentration of thrombopoietin (TPO), a key regulator of hematopoietic stem cells and megakaryocytes, revealed loss of physiologic negative correlation with platelet count in AML cases with blasts expressing MPL, the thrombopoietin (scavenging) receptor. Mechanistic studies demonstrated that MPL(hi) blasts could indeed clear TPO, likely therefore leading to insufficient cytokine levels for nonleukemic hematopoiesis. Microarray analysis in an independent multicenter study cohort of 437 AML cases validated MPL expression as a central predictor of thrombocytopenia and neutropenia in AML. Moreover, t(8;21) AML cases demonstrated the highest average MPL expression and lowest average platelet and absolute neutrophil counts among subgroups. Our work thus explains the pathophysiology of peripheral blood cytopenia in a relevant number of AML cases.

  10. Preliminary investigation of gene expression profiles in peripheral blood lymphocytes in schizophrenia.

    PubMed

    Bowden, Nikola A; Weidenhofer, Judith; Scott, Rodney J; Schall, Ulrich; Todd, Juanita; Michie, Patricia T; Tooney, Paul A

    2006-02-28

    Schizophrenia is a heterogenous disorder that is phenomenologically characterised by a combination of negative, positive, and cognitive symptoms with variable expression in the course of illness. Here, we investigated differential gene expression in relation to age to address the heterogeneity of this disorder We used 6000 gene cDNA microarrays to generate gene expression profiles from peripheral blood lymphocytes from 14 individuals with schizophrenia and 14 non-psychiatric controls. Genes showing altered expression were identified and 18 genes with brain-related functions were altered, 4 of which, endothelial differentiation gene 2 (Edg-2), ezrin-radixin-moesin phosphoprotein 50 (EBP50), Myc-associated zinc finger protein (MAZ) and Tumor Necrosis Factor Receptor 2 (TNFR2), were confirmed by relative real-time PCR. Dendrograms were constructed using genes that showed significantly different expression (p<0.05) between groups based on median split of age dividing the matched pairs into distinct subclasses. Our findings suggest that distinct gene expression profiles in peripheral blood lymphocytes associated with schizophrenia phenotypes may provide a first step towards the biological classification of schizophrenia subtypes. The validity of this approach may lead to better methods of defining this enigmatic disease.

  11. Putative Epimutagens in Maternal Peripheral and Cord Blood Samples Identified Using Human Induced Pluripotent Stem Cells

    PubMed Central

    Arai, Yoshikazu; Hayakawa, Koji; Arai, Daisuke; Ito, Rie; Iwasaki, Yusuke; Saito, Koichi; Akutsu, Kazuhiko; Takatori, Satoshi; Ishii, Rie; Hayashi, Rumiko; Izumi, Shun-Ichiro; Sugino, Norihiro; Kondo, Fumio; Horie, Masakazu; Nakazawa, Hiroyuki; Makino, Tsunehisa; Hirosawa, Mitsuko; Shiota, Kunio; Ohgane, Jun

    2015-01-01

    The regulation of transcription and genome stability by epigenetic systems are crucial for the proper development of mammalian embryos. Chemicals that disturb epigenetic systems are termed epimutagens. We previously performed chemical screening that focused on heterochromatin formation and DNA methylation status in mouse embryonic stem cells and identified five epimutagens: diethyl phosphate (DEP), mercury (Hg), cotinine, selenium (Se), and octachlorodipropyl ether (S-421). Here, we used human induced pluripotent stem cells (hiPSCs) to confirm the effects of 20 chemicals, including the five epimutagens, detected at low concentrations in maternal peripheral and cord blood samples. Of note, these individual chemicals did not exhibit epimutagenic activity in hiPSCs. However, because the fetal environment contains various chemicals, we evaluated the effects of combined exposure to chemicals (DEP, Hg, cotinine, Se, and S-421) on hiPSCs. The combined exposure caused a decrease in the number of heterochromatin signals and aberrant DNA methylation status at multiple gene loci in hiPSCs. The combined exposure also affected embryoid body formation and neural differentiation from hiPSCs. Therefore, DEP, Hg, cotinine, Se, and S-421 were defined as an “epimutagen combination” that is effective at low concentrations as detected in maternal peripheral and cord blood. PMID:26339649

  12. Effects of cadmium on metallothionein levels in human peripheral blood leukocytes: a comparison with zinc.

    PubMed

    Yurkow, E J; DeCoste, C J

    1999-11-12

    Metallothioneins (MT) are low-molecular-weight, cysteine-rich proteins that are induced in response to a variety of chemical stresses and therefore can be used to assess human exposure to environmental agents. In the current study, flow cytometry was used to characterize the basal and cadmium-induced expression of MT in the three major leukocyte populations of human peripheral blood. In the analysis, monocytes were the most sensitive leukocytes to this toxic metal, with significant increases in cellular MT levels being detected at concentrations of cadmium as low as 0.1 microM (24 h). The lymphocyte population also exhibited pronounced treatment-associated elevations in cellular MT, while the granulocyte population was found to be nonresponsive. Although both CdCl2 (3 microM) and ZnCl2 (50 microM) induced MT expression in monocytes to a similar degree and did not affect the expression of this protein in granulocytes, cadmium but not zinc treatment induced dramatic increases in MT levels of lymphocytes. Our results indicate that cellular MT protein levels, as determined by this flow cytometric method, may be used to characterize the differential responsiveness of the major human leukocyte subpopulations to transitional metals. It is evident from the current work that the responsiveness of all peripheral blood leukocyte populations should be analyzed in exposure assessment studies.

  13. Comparison of the peripheral blood leukocyte population between Japanese Black and Holstein calves.

    PubMed

    Ohtsuka, Hiromichi; Ono, Maiko; Saruyama, Yumi; Mukai, Machiko; Kohiruimaki, Masayuki; Kawamura, Seiichi

    2011-02-01

    Japanese black (JB) calves have greater susceptibility to infectious diseases compared to Holstein (Hol) calves. In order to clarify the differences in cellular immune status between JB and Hol calves, the leukocyte population and lymphocyte proliferative ability were analyzed. In total 200 healthy calves, 1 day to 14 weeks of age, were examined: 105 JB and 95 Hol calves. Lower numbers in peripheral blood and percentage in peripheral blood mononuclear cells of CD3(+)TcR1-N12(+) T cells and major histocompatibility complex class-II(+)CD14(-) B cells were observed in the JB compared to the Hol. The percentage of TcR1-N12(+)CD25(+) T cell in the JB was significantly lower than that of the Hol at 4-6, and 8-10 weeks. Interleukin (IL)-2 sensitivity in the JB was lower than that in the Hol, and significant differences were observed in age groups of 6-8 weeks and 10-14 weeks. These findings indicated that the lower numbers of γδ T cells and B cells in the JB compared to the Hol might be associated with the specificity of the immune systems in JB calves. © 2011 The Authors. Journal compilation © 2011 Japanese Society of Animal Science.

  14. Immunophenotypic and ultrastructural study in peripheral blood neutrophil granulocytes following bone marrow transplantation.

    PubMed

    Masat, T; Feliu, E; Villamor, N; Castellsagué, J; Ordi, J; Fabregues, M; Rozman, C

    1997-08-01

    Neutrophil studies after bone marrow transplantation (BMT) describe chemotactic and phagocytotic alterations and dyshaemopoiesis. Neutrophil granulocytes (NG) in peripheral blood after BMT were analysed in 28 patients. 14 patients (six receiving GM-CSF) underwent autologous BMT and 14 underwent allogeneic BMT. Immunophenotypic and electron microscopic studies were performed during post-BMT granulopoietic regeneration. Results were compared with NG from 15 healthy bone marrow donors (control group A) and from six patients receiving intensive chemotherapy before autologous BMT (control group B). A significant increase in CD15 and a decrease in 8C7 antigen expression was observed in peripheral blood NG from BMT patients compared with controls A. MPO-7 in NG after BMT did not differ from control group A. Autologous BMT patients showed a lower percentage of NG expressing 13F6, 31D8 and CD16 (Leu 11a) than allogeneic BMT patients, and a significant decrease in 8C7 antigen expression compared with patients receiving intensive chemotherapy. Ultrastructurally, a marked decrease of azurophilic granules was observed in NG from BMT patients compared with control groups A and B. These data indicate that repopulation after BMT was made by phenotypically less mature NG with dysgranulopoietic features. Differences between autologous and allogeneic BMT patients may be partly related to GM-CSF usage. In conclusion, NG present immunophenotypic and ultrastructural changes after BMT which may be involved in abnormal NG response against bacterial infections, although further investigation is needed.

  15. Monoclonal antibody (Y1/82A) with specificity towards peripheral blood monocytes and tissue macrophages.

    PubMed Central

    Davey, F R; Cordell, J L; Erber, W N; Pulford, K A; Gatter, K C; Mason, D Y

    1988-01-01

    A new monoclonal antibody, Y1/82A, was raised against phytohaemagglutinin activated peripheral blood mononuclear cells. Using an immunohistochemical technique it was shown that Y1/82A reacts against peripheral blood and bone marrow monocytes and resident macrophages from essentially all human tissues. Y1/82A bound to determinants present in leukaemic cells from patients with acute myelomonocytic leukaemia and acute monocytic leukaemia, but not to neoplastic cells from patients with malignant lymphoproliferative disorders or malignant epithelial tumours. Y1/82A failed to react with other cell types, with the exception of osteoclasts and megakaryocytes. Analysis by Western blotting showed that the antigen detected by antibody Y1/82A was associated with intracellular granules in macrophages. Monoclonal antibody Y1/82A may be useful in the diagnosis of monocytic leukaemias and histiocytic neoplasms and in the identification of macrophages in tissues from various inflammatory and neoplastic conditions. Images Fig 1-6 PMID:2457604

  16. Persistent mosaicism for 12p duplication/triplication chromosome structural abnormality in peripheral blood.

    PubMed

    Shackelford, Amy L; Conlin, Laura K; Hummel, Marybeth; Spinner, Nancy B; Wenger, Sharon L

    2013-01-01

    We present a rare case of mosaicism for a structural abnormality of chromosome 12 in a patient with phenotypic features of Pallister-Killian syndrome. A six-month-old child with dysmorphic features, exotropia, hypotonia, and developmental delay was mosaic for both a normal karyotype and a cell line with 12p duplication/triplication in 25 percent of metaphase cells. Utilization of fluorescence in situ hybridization (FISH) identified three copies of probes from the end of the short arm of chromosome 12 (TEL(12p13) locus and the subtelomere (12p terminal)) on the structurally abnormal chromosome 12. Genome-wide SNP array analysis revealed that the regions of duplication and triplication were of maternal origin. The abnormal cell line in our patient was present at 25 percent at six months and 19 months of age in both metaphase and interphase cells from peripheral blood, where typically the isochromosome 12p is absent in the newborn. This may suggest that the gene(s) resulting in a growth disadvantage of abnormal cells in peripheral blood of patients with tetrasomy 12p may not have the same influence when present in only three copies.

  17. Peripheral blood smears, bone marrow aspiration, trephine and clot biopsies: methods and protocols.

    PubMed

    Afkhami, Michelle; Vergara-Lluri, Maria; Brynes, Russell K; Siddiqi, Imran N

    2014-01-01

    Maximum diagnostic information is obtained when peripheral blood smears, bone marrow aspiration smears, trephine biopsy imprints, trephine and clot biopsy sections are simultaneously examined. Peripheral blood smears reflect end organ function and provide clues to underlying hematolymphoid pathology that may prompt additional studies including bone marrow examination. Bone marrow aspiration alone has diagnostic utility in the evaluation of a limited number of primary hematological conditions including: megaloblastic anemias, hyporegenerative anemias, certain hemolytic anemias, normochromic normocytic anemias, neutropenias, thrombocytopenias, immunoglobulin disorders, storage diseases, and leukemias (Bain, J Clin Pathol 54:657-663, 2001). Bone marrow trephine biopsy is indicated in those situations where marrow aspiration is unsuccessful; in evaluation of cytopenias, myelofibrosis, suspicion of lymphoma, metastatic tumor, granulomatous disease, evaluation of myeloproliferative neoplasms, and for the examination of trabecular bone in metabolic diseases (Bain, J Clin Pathol 54:737-742, 2001). Many of the indications for marrow aspiration overlap with those for trephine biopsy. Because it is not possible to predict which patients will have diagnostic aspiration biopsies and which will have diagnostic trephine biopsies, both procedures are routinely performed together (Brynes et al., Am J Clin Pathol 70:753-759, 1978; Cotelingam, Adv Anat Pathol 10:8-26, 2003; Lee et al., Int J Lab Hematol 30:349-364, 2008; Peterson et al., Arch Pathol Lab Med 126:1050-1056, 2002).

  18. The Effects of Impact Vibration on Peripheral Blood Vessels and Nerves

    PubMed Central

    KRAJNAK, Kristine M.; WAUGH, Stacey; JOHNSON, Claud; MILLER, G. Roger; XU, Xueyan; WARREN, Christopher; DONG, Ren G.

    2013-01-01

    Research regarding the risk of developing hand-arm vibration syndrome after exposure to impact vibration has produced conflicting results. This study used an established animal model of vibration-induced dysfunction to determine how exposure to impact vibration affects peripheral blood vessels and nerves. The tails of male rats were exposed to a single bout of impact vibration (15 min exposure, at a dominant frequency of 30 Hz and an unweighted acceleration of approximately 345 m/s2) generated by a riveting hammer. Responsiveness of the ventral tail artery to adrenoreceptor-mediated vasoconstriction and acetylcholine-mediated re-dilation was measured ex vivo. Ventral tail nerves and nerve endings in the skin were assessed using morphological and immunohistochemical techniques. Impact vibration did not alter vascular responsiveness to any factors or affect trunk nerves. However, 4 days following exposure there was an increase in protein-gene product (PGP) 9.5 staining around hair follicles. A single exposure to impact vibration, with the exposure characteristics described above, affects peripheral nerves but not blood vessels. PMID:24077447

  19. Enhanced chromosomal radiosensitivity in peripheral blood lymphocytes of larynx cancer patients

    SciTech Connect

    Lisowska, Halina; Lankoff, Anna; Wieczorek, Andrzej; Florek, Agnieszka; Kuszewski, Tomasz; Gozdz, Stanislaw; Wojcik, Andrzej . E-mail: awojcik@pu.kielce.pl

    2006-11-15

    Purpose: The chromosomal radiosensitivity in peripheral blood lymphocytes of cancer patients was reported to be higher than that of healthy donors. This effect is especially prominent when aberrations induced in the G{sub 2} phase of the cell cycle are analyzed. The aim of our study was to investigate if the G{sub 2} aberration frequencies in lymphocytes of patients with larynx cancer are higher than in the case of control individuals. Also, we tested if the frequencies of G{sub 2} aberrations correlate with side effects of radiotherapy. Methods and Materials: Peripheral blood of 38 patients was collected before the onset of radiotherapy, cultured for 72 h, and irradiated with 2 Gy after 67 h. Lymphocytes of 40 healthy donors were treated in the same way. Results: The spontaneous and radiation-induced aberration frequencies in lymphocytes of patients were on average higher than in those of healthy donors. No statistically significant correlation was observed between aberration frequencies in lymphocytes and the degree of both early and late normal tissue reactions. Conclusions: The chromosomal radiosensitivity of lymphocytes of patients with larynx cancer may be a marker of cancer predisposition; however, it does not appear to have a predictive value for the risk of developing side effects to radiotherapy.

  20. Expression profiling feline peripheral blood monocytes identifies a transcriptional signature associated with type two diabetes mellitus.

    PubMed

    O'Leary, Caroline A; Sedhom, Mamdouh; Reeve-Johnson, Mia; Mallyon, John; Irvine, Katharine M

    2017-04-01

    Diabetes mellitus is a common disease of cats and is similar to type 2 diabetes (T2D) in humans, especially with respect to the role of obesity-induced insulin resistance, glucose toxicity, decreased number of pancreatic β-cells and pancreatic amyloid deposition. Cats have thus been proposed as a valuable translational model of T2D. In humans, inflammation associated with adipose tissue is believed to be central to T2D development, and peripheral blood monocytes (PBM) are important in the inflammatory cascade which leads to insulin resistance and β-cell failure. PBM may thus provide a useful window to study the pathogenesis of diabetes mellitus in cats, however feline monocytes are poorly characterised. In this study, we used the Affymetrix Feline 1.0ST array to profile peripheral blood monocytes from 3 domestic cats with T2D and 3 cats with normal glucose tolerance. Feline monocytes were enriched for genes expressed in human monocytes, and, despite heterogeneous gene expression, we identified a T2D-associated expression signature associated with cell cycle perturbations, DNA repair and the unfolded protein response, oxidative phosphorylation and inflammatory responses. Our data provide novel insights into the feline monocyte transcriptome, and support the hypothesis that inflammatory monocytes contribute to T2D pathogenesis in cats as well as in humans. Copyright © 2017 Elsevier B.V. All rights reserved.

  1. The Role of ABO Incompatibility in Allogeneic Peripheral Blood Stem Cell Transplant.

    PubMed

    Arslan, Önder; Coşkun, Hasan Şanol; Arat, Mutlu; Soydan, Ender; Özcan, Muhit; Gürman, Günhan; Çelebi, Harika; Demirer, Taner; Akan, Hamdi; İlhan, Osnman; Konuk, Nahide; Uysal, Akın; Berksaç, Meral; Koç, Haluk

    2002-09-05

    ABO incompatibility is not a contraindication for allogeneic bone marrow transplantation, but this procedure requires an extra effort for erythrocyte or plasma depletion in certain well established conditions. Some acute or delayed immunohematological complications such as acute or chronic hemolysis and pure red cell aplasia may be encountered. In this study the outcome and transplant related complications of ABO incompatible and identical cases, who have received allogeneic peripheral blood stem cells from their HLA identical siblings were compared with each other. Ninety-one patients (CML 36, AML 37, other 18) were analyzed retrospectively including 51 (60.4%) ABO identical patients and 36 (39.6%) ABO mismatched (MM) patients, who have a bi-directional MM (n= 5), major MM (n= 16), minor MM (n= 9) and Rh MM (n= 6). Median follow up was 13 (0.5-43.0) months. We did not observed any significant differences between two groups (identical vs non-identical) in terms of acute hemolysis preceding stem cell infusion, peritransplant transfusion demand, acute- and chronic graft versus host disease. There was no change in estimated disease free survival and overall survival durations. We did not observed any influence of ABO/Rh incompatibility on short term outcome in allogeneic peripheral blood stem cell transplantation in our series and did not recommend further manipulation of the infused stem cells.

  2. A gene expression signature of confinement in peripheral blood of red wolves (Canis rufus).

    PubMed

    Kennerly, Erin; Ballmann, Anne; Martin, Stanton; Wolfinger, Russ; Gregory, Simon; Stoskopf, Michael; Gibson, Greg

    2008-06-01

    The stresses that animals experience as a result of modification of their ecological circumstances induce physiological changes that leave a signature in profiles of gene expression. We illustrate this concept in a comparison of free range and confined North American red wolves (Canis rufus). Transcription profiling of peripheral blood samples from 13 red wolf individuals in the Alligator River region of North Carolina revealed a strong signal of differentiation. Four hundred eighty-two out of 2980 transcripts detected on Illumina HumanRef8 oligonucleotide bead arrays were found to differentiate free range and confined wolves at a false discovery rate of 12.8% and P < 0.05. Over-representation of genes in focal adhesion, insulin signalling, proteasomal, and tryptophan metabolism pathways suggests the activation of pro-inflammatory and stress responses in confined animals. Consequently, characterization of differential transcript abundance in an accessible tissue such as peripheral blood identifies biomarkers that could be useful in animal management practices and for evaluating the impact of habitat changes on population health, particularly as attention turns to the impact of climate change on physiology and in turn species distributions.

  3. Increased numbers of peripheral blood CD34+ cells in dogs with canine atopic dermatitis.

    PubMed

    Bruet, Vincent; Lieubeau, Blandine; Herve, Julie; Roussel, Anne; Imparato, Laëtitia; Desfontis, Jean-Claude; Bourdeau, Patrick

    2015-06-01

    The bone marrow may be involved in human atopic diseases, as shown by the release of CD34+ cells into the peripheral blood. The aim was to determine the numbers of CD34+ cells in atopic dogs. The following three groups of dogs were studied: 27 dogs with nonfood-induced atopic dermatitis (NFICAD); 16 dogs with nonallergic inflammatory diseases; and 13 healthy control dogs. Dogs with NFICAD were selected after fulfilment of Favrot's criteria and exclusion of other pruritic dermatoses, including flea infestation and adverse reaction to foods. The Canine Atopic Dermatitis Extent and Severity Index (CADESI)-03 and a Visual Analog Scale (VAS) score for pruritus were used to quantify clinical signs. A phycoerythrin-conjugated anticanine CD34 antibody was used to stain peripheral blood CD34+ cells, and these were enumerated using a flow cytometer. The CD34+ cell counts were compared between groups and tested (in the NFICAD group) for correlation with the severity of clinical signs. The numbers of peripheral CD34+ cells in dogs with NFICAD (median 1.7) were statistically higher than in dogs with other nonallergic inflammatory diseases (median 1.0; P = 0.01) and healthy control dogs (median 0.9; P = 0.009). In dogs with NFICAD, there was no correlation between CD34+ cell numbers and CADESI-03 scores or owner-assessed pruritus (VAS score). The results of this study suggest the possible involvement of CD34+ cells in dogs with NFICAD. The role of CD34+ cells in the aetiopathogenesis of canine atopic dermatitis remains to be determined. © 2014 ESVD and ACVD.

  4. Signature of subclinical femoral artery atherosclerosis in peripheral blood mononuclear cells.

    PubMed

    Llorente-Cortés, Vicenta; de Gonzalo-Calvo, David; Orbe, Josune; Páramo, Jose Antonio; Badimon, Lina

    2014-06-01

    Peripheral arterial disease is a relevant public health problem associated with increased risk of morbimortality. Most of the patients with this condition are asymptomatic. Therefore, the development of accessible biochemical markers seems to be necessary to anticipate diagnosis. Our hypothesis is that asymptomatic subjects with objectively confirmed femoral artery atherosclerosis could be distinguished from control subjects by gene expression analysis in peripheral blood mononuclear cells (PBMC). A total of 37 asymptomatic males over 50 years old were recruited at the University Clinic of Navarra (Spain). Nineteen participants were free from atherosclerotic vascular disease and 18 participants presented subclinical femoral artery atherosclerosis defined by means of Doppler ultrasound. PBMC were isolated from blood and the RNA extracted. A panel of atherosclerotic-related genes were evaluated by Taqman low-density array. In univariate logistic regression models, we found a direct relationship between IL4, ITGAM and TLR2 expression levels in PBMC and femoral atherosclerosis, even when the models were adjusted for age and hypertension prevalence. Multivariate logistic regression models showed that elevated IL4 expression levels were intimately associated with subclinical femoral atherosclerosis after adjusting for the same potential confounders. Current data suggest that gene expression in PBMC, in particular IL4 expression, could be a useful tool in the diagnosis of femoral artery atherosclerosis in asymptomatic patients. Furthermore, in patients with no differences in cardiovascular risk factors except for hypertension, the results point to the immune and inflammatory deregulation as a feature of subclinical peripheral atherosclerosis. © 2014 Stichting European Society for Clinical Investigation Journal Foundation.

  5. Peripheral venous distension elicits a blood pressure raising reflex in young and middle-aged adults.

    PubMed

    Matthews, Evan L; Brian, Michael S; Coyle, Dana E; Edwards, David G; Stocker, Sean D; Wenner, Megan M; Farquhar, William B

    2016-06-01

    Distension of peripheral veins in humans elicits a pressor and sympathoexcitatory response that is mediated through group III/IV skeletal muscle afferents. There is some evidence that autonomic reflexes mediated by these sensory fibers are blunted with increasing age, yet to date the venous distension reflex has only been studied in young adults. Therefore, we tested the hypothesis that the venous distension reflex would be attenuated in middle-aged compared with young adults. Nineteen young (14 men/5 women, 25 ± 1 yr) and 13 middle-aged (9 men/4 women, 50 ± 2 yr) healthy normotensive participants underwent venous distension via saline infusion through a retrograde intravenous catheter in an antecubital vein during limb occlusion. Beat-by-beat blood pressure, muscle sympathetic nerve activity (MSNA), and model flow-derived cardiac output (Q), and total peripheral resistance (TPR) were recorded throughout the trial. Mean arterial pressure (MAP) increased during the venous distension in both young (baseline 83 ± 2, peak 94 ± 3 mmHg; P < 0.05) and middle-aged adults (baseline 88 ± 2, peak 103 ± 3 mmHg; P < 0.05). MSNA also increased in both groups [young: baseline 886 ± 143, peak 1,961 ± 242 arbitrary units (AU)/min; middle-aged: baseline 1,164 ± 225, peak 2,515 ± 404 AU/min; both P < 0.05]. TPR (P < 0.001), but not Q (P = 0.76), increased during the trial. However