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Sample records for lymphocyte culture mlc

  1. [Computer-assisted histocompatibility assessment in mixed lymphocyte culture (MLC)].

    Science.gov (United States)

    Schwartz, D; Hajek-Rosenmayr, A

    1987-02-20

    Analysis of the results of mixed lymphocyte culture (MLC) for compatibility testing preceding transplantation of bone marrow and other organs has so far required a vast input, both in terms of laboratory staff and work hours. We have developed a computer programme which performs this work rapidly. Graphics of the reaction patterns can be obtained, moreover, and these can prove a helpful tool in interpretation of the results.

  2. Proteolytically modified human beta 2-microglobulin augments the specific cytotoxic activity in murine mixed lymphocyte culture

    DEFF Research Database (Denmark)

    Nissen, Mogens Holst; Claësson, M H

    1987-01-01

    (M-beta 2-m) bind to murine lymphocytes expressing H-2 class I antigens; M-beta 2-m, when added at day 0 and 1 of culture in nanomolar concentrations to a one-way murine allogeneic mixed lymphocyte culture (MLC) augments the generation of specific cytotoxic T lymphocytes; M-beta 2-m increases...... the endogenous production of interleukin 2 in the MLC culture; monoclonal antibody which reacts with both the native beta 2-m and M-beta 2-m molecule blocks the augmentation of cytotoxic T lymphocyte production induced by M-beta 2-m; murine as well as human MLC responder cells can proteolytically modify native...

  3. Limiting dilution analysis of alloantigen-reactive cells which respond to allogeneic lymphocytes in human MLC and PLT.

    Science.gov (United States)

    Singal, D P; Naipaul, N; Joseph, S

    1980-10-01

    We have determined the frequency of the alloantigen-reactive cells (ARC) in human MLC and PLT by the limiting dilution analysis. In PLT, the frequency of the ARC to the original sensitizing donor ranged between 1:32 to 1:62, an increase of six to nine-fold after priming in MLC. The MLC primed populations were also enriched (three to five fold) for the ARC responding to the PL-positive allogeneic donors. The incidence of the ARC was 1:62 to 1:118 with donors positive for the sensitizing HLA-DRw antigen and 1:77 to 1:144 with donors negative for the specific HLA-DRw determinant. The results from experiments utilizing pooled stimulating cells from the original and allogeneic donors suggest that same subpopulation of cells responds to the sensitizing HLA-DRw determinant, whether it is presented by the specific stimulator or by a third-party allogeneic donor. On the other hand, different subpopulations of alloreactive cells respond to different alloantigens. In MLC experiments between HLA-identical siblings, the incidence of the ARC ranged between 1:995 to 1:1673. The responses of the ARC to non-HLA antigens were observed under conditions where responder cells were limiting. Also, the responses of the limiting numbers of responding cells were inhibited by mitomycin-treated autologous lymphocytes. Nonresponse in MLC combinations with higher responder cell numbers was not due to small numbers of stimulating cells.

  4. Typing for MLC determinants using lymphocytes from cousin-marriage offspring as typing cells.

    Science.gov (United States)

    Singal, D P; Villanueva, R; Naipaul, N

    1975-07-01

    A first cousin marriage family with three children was studied. One sibling (AW) was homozygous for W32.W10. When used as stimulator in one way MLC, AW elicited a very low response in family members possessing this haplotype. Serologically identical sibling and father combination showed bidirectional non-stimulation in one way MLC. Therefore, AW is assumed to be homozygous not only for SD antigens, but also for LD determinants which we call LD W10a. Among the 43 unrelated individuals tested with AW, an association between LD W10a and W10 antigen of the second HL-A locus was observed. Of 12 W10 positive individuals tested, five carry this allele as compared to one out of 31 W10 negative individuals. The data suggest a strong linkage disequilibrium between the FOUR locus determinant W10 and LD determinant LD W10a.

  5. Interactions of allogeneic human mononuclear cells in the two-way mixed leucocyte culture (MLC): influence of cell numbers, subpopulations and cyclosporin

    Science.gov (United States)

    Sato, T; Deiwick, A; Raddatz, G; Koyama, K; Schlitt, H J

    1999-01-01

    With organ allografts considerable numbers of donor-type mononuclear cells are transferred to the recipient, leading to bilateral immunological interactions between donor and recipient lymphocytes. To study such bilateral immune reactions in detail, human two-way MLC were performed. In this model proliferation kinetics, patterns of activation, and survival of the two populations were analysed, and the relevance of initial cell subset composition, relative cell numbers, and the effect of immunosuppression on this co-culture were evaluated. It could be demonstrated that with an initial 50:50 ratio of two populations of allogeneic cells one population dominated after 21 days of co-culture in 78 out of 80 combinations (97%) tested; the other population decreased markedly after an initially stable phase of 6–7 days. With unequal starting conditions the larger population dominated when resting cells were used, but small populations of preactivated cells or separated CD8+ cells could also dominate. Depletion of CD16+ natural killer (NK) cells and of CD2− cells (B cell and monocytes) had no effect on domination. Addition of cyclosporin delayed or blocked the domination process while addition of IL-2 accelerated it. Disappearance of one population was associated with detection of apoptotic cells. The findings indicate that co-cultures of allogeneic mononuclear cells are generally not stable for more than 1 week, but lead to active elimination of one population. CD8+ cells and particularly preactivated cells seem to play the most important role in that process, while NK cells are of less importance. Cyclosporin can prolong survival of allogeneic cells in co-culture. These observations suggest that under the conditions of clinical organ transplantation even small amounts of immunocompetent donor cells transferred by the graft may persist for some time and may, thereby, have the chance to exert immunomodulatory functions. PMID:9933457

  6. Selective effects of alpha interferon on human T-lymphocyte subsets during mixed lymphocyte cultures

    DEFF Research Database (Denmark)

    Hokland, M; Hokland, P; Heron, I

    1983-01-01

    Mixed lymphocyte reaction (MLR) cultures of human lymphocyte subsets with or without the addition of physiological doses of human alpha interferon (IFN-alpha) were compared with respect to surface marker phenotypes and proliferative capacities of the responder cells. A selective depression on the T...

  7. Transfer of cholesterol from macrophages to lymphocytes in culture.

    Science.gov (United States)

    de Bittencourt Júnior, P I; Curi, R

    1998-02-01

    A major feature of macrophage metabolism is its capacity to produce and export cholesterol. Several reports have shown that the manipulation of lymphocyte cholesterol content elicits important changes in lymphocyte proliferation. These findings lead to an inquiry as to whether macrophage-derived cholesterol released into the lymphocyte surroundings may be transferred to the latter thus affecting lymphocyte function. In this study, cholesterol transfer from macrophages to lymphocytes was examined in vitro using rat cells in culture. The findings indicate that there may be a significant transfer of cholesterol from [4-14C]cholesterol labeled resident peritoneal macrophages to mesenteric lymph node resting lymphocytes (up to 173.9 +/- 2.7 pmol/10(7) lymphocytes/10(7) macrophages when co-cultivated for 48 h), in a lipoprotein-dependent manner. This represents the mass transfer of ca. 17 nmoles of cholesterol molecules per 10(7) lymphocytes from 10(7) macrophages (calculated on the basis of specific radioactivity incorporated into macrophages after the pre-labelling period), which suggests that macrophages are capable of replacing the whole lymphocyte cholesterol pool every 21 h. Moreover, an 111%-increase in the total cholesterol content of lymphocytes was found after co-cultivation with macrophages for 48 h. When compared to peritoneal cells, monocytes/macrophages obtained from circulating blood leukocytes presented a much higher cholesterol transfer capacity to lymphocytes (3.06 +/- 0.10 nmol/10(7) lymphocytes/10(7) macrophages co-cultivated for 24 h). Interestingly, inflammatory macrophages dramatically reduced their cholesterol transfer ability (by up to 91%, as compared to resident macrophages). Cholesterol transfer may involve a humoral influence, since it is not only observed when cells are co-cultivated in a single-well chamber system (cells in direct contact), but also in a two-compartment system (where cells can communicate but not by direct contact). Co

  8. Genotoxicity of the herbicide butachlor in cultured human lymphocytes.

    Science.gov (United States)

    Sinha, S; Panneerselvam, N; Shanmugam, G

    1995-08-01

    Butachlor, a pre-emergence herbicide was investigated for its ability to induce sister chromatid exchanges (SCE) and chromosome aberrations (CA) in cultured human peripheral blood lymphocytes. Mitogen-stimulated lymphocytes were treated with three different concentrations (5, 10 and 20 micrograms/ml) of butachlor for 24, 48 and 72 h. Our results indicate a dose-dependent increase in the frequency of chromosomal aberrations at 24, 48 and 72 h of treatment with butachlor. No SCE was promoted by butachlor.

  9. Growing B Lymphocytes in a Three-Dimensional Culture System

    Science.gov (United States)

    Wu, J. H. David; Bottaro, Andrea

    2010-01-01

    A three-dimensional (3D) culture system for growing long-lived B lymphocytes has been invented. The capabilities afforded by the system can be expected to expand the range of options for immunological research and related activities, including testing of immunogenicity of vaccine candidates in vitro, generation of human monoclonal antibodies, and immunotherapy. Mature lymphocytes, which are the effectors of adaptive immune responses in vertebrates, are extremely susceptible to apoptotic death, and depend on continuous reception of survival-inducing stimulation (in the forms of cytokines, cell-to-cell contacts, and antigen receptor signaling) from the microenvironment. For this reason, efforts to develop systems for long-term culture of functional, non-transformed and non-activated mature lymphocytes have been unsuccessful until now. The bone-marrow microenvironment supports the growth and differentiation of many hematopoietic lineages, in addition to B-lymphocytes. Primary bone-marrow cell cultures designed to promote the development of specific cell types in vitro are highly desirable experimental systems, amenable to manipulation under controlled conditions. However, the dynamic and complex network of stromal cells and insoluble matrix proteins is disrupted in prior plate- and flask-based culture systems, wherein the microenvironments have a predominantly two-dimensional (2D) character. In 2D bone-marrow cultures, normal B-lymphoid cells become progressively skewed toward precursor B-cell populations that do not retain a normal immunophenotype, and such mature B-lymphocytes as those harvested from the spleen or lymph nodes do not survive beyond several days ex vivo in the absence of mitogenic stimulation. The present 3D culture system is a bioreactor that contains highly porous artificial scaffolding that supports the long-term culture of bone marrow, spleen, and lymph-node samples. In this system, unlike in 2D culture systems, B-cell subpopulations developing

  10. C1-esterase inhibitor blocks T lymphocyte proliferation and cytotoxic T lymphocyte generation in vitro

    DEFF Research Database (Denmark)

    Nissen, Mogens Holst; Bregenholt, S; Nording, J A

    1998-01-01

    We have previously shown that activated C1s complement and activated T cells cleave beta2-microglobulin (beta2m) in vitro leading to the formation of desLys58 beta2m. This process can specifically be inhibited by C1-esterase inhibitor (C1-inh). Furthermore we showed that exogenously added desLys58...... beta2m in nanomolar amounts to a one-way allogenic mixed lymphocyte culture (MLC) increased the endogenous production of IL-2 and the generation of allo-specific cytotoxic T lymphocytes. C1-inh was purified from fresh human plasma and added to human or murine MLC and mitogen-stimulated lymphocyte...... of allospecific cytotoxic activity, and changed the endogenous production of IL-2, IL-4, IL-10, IL-12 and IFN-gamma. These data clearly demonstrate a regulatory function of C1-inh on T cell-mediated immune functions....

  11. Light microscope observation of circulating human lymphocytes cultured in vitro

    Directory of Open Access Journals (Sweden)

    Naila Francis Paulo de Oliveira

    2010-10-01

    Full Text Available The purpose of this work was to study the isolation and a light microscopy technique for cultured lymphocytes. Blood samples were obtained by venipuncture with an anticoagulant added and centrifuged in a Percoll density gradient to separate the leukocytes. Lymphocytes were placed in 25 cm ³ tissue culture flasks at 37ºC. After culturing, they were fixed and stained with the methods used for blood smears. Results showed that not all fixing solutions and stains were an equally good choice for cultured lymphocytes.Os linfócitos são células importantes do sistema imune e têm sido largamente utilizados em estudos morfológicos. Entretanto, a literatura sobre técnicas de preparação dessas células é escassa e antiga, especialmente para linfócitos cultivados in vitro. Portanto, o objetivo desse estudo foi relatar com detalhes as técnicas de isolamento e microscopia de luz de linfócitos mantidos em cultura. Amostras de sangue foram obtidas por punção venosa e centrifugadas em gradiente de densidade de Percoll, para separar os leucócitos. Os linfócitos foram mantidos em frascos de cultura de 25 cm³ a 37ºC. Após a cultura, as células foram fixadas e coradas de acordo com a metodologia utilizada para esfregaços sanguíneos. Nossos resultados mostraram que nem todos os fixadores e corantes utilizados para esfregaços sanguíneos são uma boa escolha para linfócitos cultivados in vitro.

  12. Analysis of Delphinidin and Luteolin Genotoxicity in Human Lymphocyte Culture

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    Jasmin Ezić

    2015-08-01

    Full Text Available Introduction: Bioflavonoids delphinidin (2-(3,4,5-Trihydroxyphenylchromenylium-3,5,7-triol and luteolin (2-(3,4-Dihydroxyphenyl-5,7-dihydroxy-4-chromenone have been recognized as promising antioxidants and anticancer substances. Due to their extensive use, the goal of the research was to determine whether they have any genotoxic potential in vitro.Methods: Analysis of genotoxic potential was performed applying chromosome aberrations test in human lymphocyte culture, as this kind of research was not conducted abundantly for these two bioflavonoids. Delphinidin and luteolin were dissolved in DMSO and added to cultures in final concentrations of 25, 50 and 100 μM.Results: In human lymphocytes cultures Delphinidin induced PCDs in all treatments, potentially affecting the cell cycle and topoisomerase II activity. In concentration of 50 μM luteolin showed strong genotoxic effects and caused significant reduction of cell proliferation.Conclusion: Luteolin exhibited certain genotoxic and cytostatic potential. Delphinidin was not considered genotoxic, however its impact on mitosis, especially topoisomerase II activity, was revealed.

  13. Influence of a constant magnetic field on human lymphocyte cultures.

    Science.gov (United States)

    Ardito, G; Lamberti, L; Bigatti, P; Prono, G

    1984-07-31

    The growing exposure to magnetic fields of a certain intensity could represent a serious hazard for our health. In the present note we analyze the effect of a 740 Gauss magnetic field on human lymphocyte cell cultures. From the analysis of our data it is possible to point out that this field produces an inhibition of the cell growth, while does not affect at all the sister chromatid exchange frequency of the chromosomes. Conversely we found a significant increase of chromosome aberrations in the exposed cells. The chromosome aberrations found were mostly gaps and breaks.

  14. Cytogenetic effects of the herbicide chloridazon in cultured sheep lymphocytes.

    Science.gov (United States)

    Siviková, K; Dianovský, J

    1997-01-01

    The effect of in vitro exposure to the herbicide chloridazon on the induction of chromosome aberrations (CA) and sister chromatid exchanges (SCE) was studied in cultured sheep peripheral lymphocytes. A positive clastogenic effect was observed in chloridazon-treated cultures at a dose of 7 x 10(-4) M both in the presence and absence of the S9 fraction, but no significant increase of chromosome breaks was seen at lower doses (7 x 10(-6) M and 7 x 10(-5) M, respectively). A clear dose-dependence and significant differences were found in chloridazon potency to induce SCEs. Induction of cell cycle delays as compared to the controls was not observed.

  15. Suppressive effects of antigens on the activity of specific activated lymphocytes: A test to define the specificity of activated lymphocytes

    Institute of Scientific and Technical Information of China (English)

    HU Jun; PAN Sheng-jun; CAI Zhen-jie; GUAN De-lin; LIU Xiao-cheng

    2006-01-01

    Objective:With the regular mixed lymphocytes culture (MLC) to detect the allograft rejection, the reactivity of the activated lymphocytes (primed lymphocytes) of a recipient shows sometimes increase and sometimes decrease against the antigens from the donor, which is inconsistent with the clinical results. In order to establish a convenient method for testing the specificity of the activated lymphocytes in vitro, so as to know the rejection occurred or not by testing the existence of the specific activated lymphocytes against donor's HLA antigens in the recipient's peripheral blood. Methods: Anti-IL-2 neutralizing monoclonal antibody (anti-IL-2 N-mAb) and immunosuppressors were introduced in this test system in the presence of specific stimulators and activated lymphocytes. Results: When the activated lymphocytes were chosen from the one-way MLC 4 d to undergo re-stimulation by specific stimulators, the activity of activated lymphocytes in the treatment group was suppressed significantly compared with that in the control group. The result of this test method is consistent with the biopsy in the clinical diagnosis of rejection.Conclusion :It suggests that the activated lymphocytes can be inactivated by specific antigens in certain conditions. This can be a useful tool to define the specificity of the activated lymphocytes.

  16. T-cell activation. VI. Inhibitory and stimulatory effects of anti-major histocompatibility complex class I antibodies in allogeneic mixed lymphocyte culture

    DEFF Research Database (Denmark)

    Röpke, M; Röpke, C; Claesson, Mogens Helweg

    1993-01-01

    Murine T splenocytes stimulated in primary allogeneic mixed lymphocyte culture (MLC) were incubated with soluble anti-major histocompatibility complex (MHC) class I monoclonal antibodies. These antibodies induced inhibition in the cytotoxicity of the responding population and this inhibition...... was not dependent on the domain on class I molecules recognized by the antibodies. Cross-reactivity of the antibodies between the responder and stimulating cell population caused a marked reduction in the inhibitory effect compared to systems where no such cross-reactivity was present. Saturating levels...... of the antibodies caused a reduction in generation of T-cell cytotoxicity, whereas low concentrations stimulated the same response. These results demonstrate that the MHC class I molecules of T cells are of significant importance in antigen-induced signal transduction....

  17. Virtual micro MLC commissioning.

    Science.gov (United States)

    Drever, Laura; Dickof, Peter

    2005-01-01

    The resolution of multileaf collimators (MLCs) is limited by their finite leaf width. A commercial package (HD-270) uses 3D couch translation and leaf adjustments to emulate smaller leaf widths. In this paper, we report on the commissioning of this feature using software testing, dial gauge measurements, and film dosimetry. We also identify a variety of limitations: software bugs and truncation artifacts, MLC leaf positioning uncertainties (random variations, systematic gantry dependence and backlash), and uncertainties in couch positioning. These reduce the capabilities of this implementation below that achievable theoretically.

  18. Basic Principles of MLC

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    Maria Rambla-Alegre

    2012-01-01

    Full Text Available Micellar liquid chromatography (MLC is an efficient alternative to conventional reversed-phase liquid chromatography with hydro-organic mobile phases. Almost three decades of experience have resulted in an increasing production of analytical applications. Current concern about the environment also reveals MLC as an interesting technique for “green” chemistry because it uses mobile phases containing 90% or more water. These micellar mobile phases have a low toxicity and are not producing hazardous wastes. The stationary phase is modified with an approximately constant amount of surfactant monomers, and the solubilising capability of the mobile phase is altered by the presence of micelles, giving rise to a great variety of interactions (hydrophobic, ionic, and steric with major implications in retention and selectivity. From its beginnings in 1980, the technique has evolved up to becoming in a real alternative in some instances (and a complement in others to classical RPLC with aqueous-organic mixtures, owing to its peculiar features and unique advantages. The addition of an organic solvent to the mobile phase was, however, soon suggested in order to enhance the low efficiencies and weak elution strength associated with the mobile phases that contained only micelles.

  19. New chicken Rfp-Y haplotypes on the basis of MHC class II RFLP and MLC analyses

    DEFF Research Database (Denmark)

    Juul-Madsen, H R; Zoorob, R; Auffray, C;

    1997-01-01

    New chicken Rfp-Y haplotypes were determined by the use of restriction fragment length polymorphism (RFLP) and mixed lymphocyte culture (MLC) in four different chicken haplotypes, B15, B19, B21, B201. The RFLP polymorphism was mapped to the Rfp-Y system by the use of a subclone (18.1) which maps...... near a polymorphic lectin gene located in the Rfp-Y system and DNA from families with known segregation of the implicated RFLP polymorphism. For the first time it is shown that major histocompatibility complex class II genes in the Rfp-Y system have functional implications. Sequence information...

  20. Expression of an accessory cell phenotype by hairy cells during lymphocyte colony formation in agar culture.

    Science.gov (United States)

    Farcet, J P; Gourdin, M F; Testa, U; Andre, C; Jouault, H; Reyes, F

    1983-01-01

    Human T lymphocytes require the cooperation of accessory cells to generate lymphocyte colonies in agar culture under PHA stimulation. Various hairy cell enriched fractions, as well as normal monocytes, have been found to be able to initiate colony formation by normal lymphocytes. Leukemic monocytes from CMML patients were also effective, but not the leukemic lymphocytes from CLL patients. The phenotype expressed by HC in agar colonies was further studied using cell surface and enzymatic markers. We have concluded that HC in agar culture in the presence of both normal T lymphocytes and PHA lose the B phenotype that they express in vivo and function like an accessory cell in contrast to normal or leukemic B lymphocytes.

  1. Cultured lymphocytes from alcoholic subjects have altered cAMP signal transduction.

    OpenAIRE

    Nagy, L E; Diamond, I; Gordon, A.

    1988-01-01

    Previous work has shown that freshly isolated lymphocytes from alcoholic subjects show significantly reduced basal and adenosine receptor-stimulated cAMP levels. This decrease could be due to ethanol-induced cellular adaptation or to a genetic difference in the regulation of cAMP signal transduction. Therefore, we cultured human lymphocytes in defined medium without ethanol for 7-8 days and then examined differences in receptor-dependent cAMP accumulation between lymphocytes from alcoholic an...

  2. RhoA-mediated MLC2 regulates actin dynamics for cytokinesis in meiosis.

    Science.gov (United States)

    Duan, Xing; Liu, Jun; Zhu, Cheng-Cheng; Wang, Qiao-Chu; Cui, Xiang-Shun; Kim, Nam-Hyung; Xiong, Bo; Sun, Shao-Chen

    2016-01-01

    During oocyte meiosis, the bipolar spindle forms in the central cytoplasm and then migrates to the cortex. Subsequently, the oocyte extrudes the polar body through two successive asymmetric divisions, which are regulated primarily by actin filaments. Myosin light chain2 (MLC2) phosphorylation plays pivotal roles in smooth muscle contraction, stress fiber formation, cell motility and cytokinesis. However, whether MLC2 phosphorylation participates in the oocyte polarization and asymmetric division has not been clarified. The present study investigated the expression and functions of MLC2 during mouse oocyte meiosis. Our result showed that p-MLC2 was localized in the oocyte cortex, with a thickened cap above the chromosomes. Meanwhile, p-MLC2 was also localized in the poles of spindle. Disruption of MLC2 activity by MLC2 knock down (KD) caused the failure of polar body extrusion. Immunofluorescent staining showed that a large proportion of oocytes arrested in telophase stage and failed to undergo cytokinesis after culturing for 12 hours. In the meantime, actin filament staining at oocyte membrane and cytoplasm were reduced in MLC2 KD oocytes. Finally, we found that the phosphorylation of MLC2 protein levels was decreased after disruption of RhoA activity. Above all, our data indicated that the RhoA-mediated MLC2 regulates the actin organization for cytokinesis during mouse oocyte maturation.

  3. Comparison of the effects of polysaccharides from wood-cultured and bag-cultured Ganoderma lucidum on murine spleen lymphocyte proliferation in vitro%段木栽培及袋栽灵芝多糖对体外培养小鼠脾淋巴细胞增殖活性的比较

    Institute of Scientific and Technical Information of China (English)

    曹立珍; 林志彬

    2003-01-01

    目的比较段木栽培灵芝多糖(wood-cultured Ganoderma lucidum polysaccharides, Gl-PS-WC) 及袋栽灵芝多糖(bag-cultured Ganoderma lucidum polysaccharides, Gl-PS-BC)对体外培养小鼠脾淋巴细胞增殖活性的影响,探讨袋栽灵芝多糖替代段木栽培灵芝多糖的可能性. 方法检测两种灵芝多糖对混合淋巴细胞培养(MLC)反应的影响;观察对刀豆蛋白A (Con A)、细菌脂多糖(LPS)诱导淋巴细胞增殖的影响以及对环孢素A (CsA)、丝裂霉素C(Mit C)、足叶乙苷(VP-16) 等抑制MLC反应的影响.结果当质量浓度为0.2~12.8 mg*L-1时,两种灵芝多糖均可促进MLC反应,增强Con A或LPS诱导的淋巴细胞增殖,并拮抗CsA, Mit C或VP-16对MLC反应的抑制作用.未发现两种多糖之间有显著性差异.结论 Gl-PS-WC及Gl-PS-BC对体外培养脾淋巴细胞的增殖活性有类似作用.%Aim To compare the influences of wood-cultured Ganoderma lucidum polysaccharides (Gl-PS-WC) and bag-cultured Ganoderma lucidum polysaccharides (Gl-PS-BC) on the proliferation activities of murine spleen lymphocytes in vitro, and investigate whether Gl-PS-BC can be substituted for Gl-PS-WC. Methods Mixed lymphocyte culture (MLC) reaction, lymphocyte proliferation induced by concanavalin A (Con A, 1 mg*L-1) or lipopolysaccharide (LPS, 5 mg*L-1), MLC reactions inhibited by immunosuppressive drugs, cyclosporine A (CsA, 0.1 mg*L-1), mitomycin (Mit C, 0.1 mg*L-1), or antitumor drug, etoposide (VP-16, 0.1 mg*L-1), were detected in the presence or absence of Gl-PS-WC and Gl-PS-BC in the concentration range of 0.2-12.8 mg*L-1. Results Two kinds of polysaccharides were shown to promote MLC in the range of 0.2-12.8 mg*L-1, increase lymphocyte proliferation induced by Con A or LPS and antagonize the inhibitory effects of CsA, Mit C or VP-16 on MLC. No significant difference was observed between these two kinds of polysaccharides in selected concentrations. Conclusion Gl-PS-WC and Gl-PS-BC showed similar effects

  4. Analysis of IL-2-like factor in lymphocyte culture supernatant of olive flounder, Paralichthys oliveaceus

    Institute of Scientific and Technical Information of China (English)

    WU Riqin; ZHANG Peijun; LI Jun; XU Yongli

    2005-01-01

    To study immune mechanism of fish lymphocyte we performed a proliferation assay and ELISA using monoclonal antibody against human IL-2. The result showed that an interleukin-2 (IL-2)-like factor was detected in the supernatant of plant haemoglutinin (PHA)-stimulated lymphocyte culture from peripheral blood,spleen and head kidney of olive flounder, Paralichthys olivaceus. The quantities of IL-2-1ike factor in the supematant from different lymphoid tissues were quite different. The IL-2 like factor in the supernatant from cultured head kidney lymphocytes was much higher than those of peripheral blood lymphocytes and spleen lymphocytes (P<0.01). The IL-2 activity was found in either mouse thymocyte proliferation assay or flounder head kidney lymphocyte proliferation assay and shown to have obvious enhancing effect on proliferation of the above two types of cell. The recombinant human IL-2 (rhIL-2) was able to stimulate flounder thymocyte proliferation and used to detect the IL-2 receptor (IL-2R) on the surface of flounder lymphocyte. The cross-reaction between the lymphocytes of flounder peripheral blood and CD25(IL-2R) was detected with flow cytometry and shown that the percentage of CD25-positive cell in peripheral blood was 7.74± 0.67%.

  5. Addition of exogenous cytokines in mixed lymphocyte culture for selecting related donors for bone marrow transplantation

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    Jeane Eliete Laguila Visentainer

    Full Text Available CONTEXT: Mixed lymphocyte culturing has led to conflicting opinions regarding the selection of donors for bone marrow transplantation. The association between a positive mixed lymphocyte culture and the development of graft-versus-host disease (GVHD is unclear. The use of exogenous cytokines in mixed lymphocyte cultures could be an alternative for increasing the sensitivity of culture tests. OBJECTIVE: To increase the sensitivity of mixed lymphocyte cultures between donor and recipient human leukocyte antigen (HLA identical siblings, using exogenous cytokines, in order to predict post-transplantation GVHD and/or rejection. TYPE OF STUDY: Prospective study. SETTING: Bone Marrow Transplantation Unit, Universidade Estadual de Campinas. PARTICIPANTS: Seventeen patients with hematological malignancies and their respective donors selected for bone marrow transplantation procedures. PROCEDURES: Standard and modified mixed lymphocyte culturing by cytokine supplementation was carried out using donor and recipient cells typed for HLA. MAIN MEASUREMENTS: Autologous and allogenic responses in mixed lymphocyte cultures after the addition of IL-4 or IL-2. RESULTS: In comparison with the standard method, average responses in the modified mixed lymphocyte cultures increased by a factor of 2.0 using IL-4 (p < 0.001 and 6.4 using IL-2 (p < 0.001, for autologous donor culture responses. For donor-versus-recipient culture responses, the increase was by a factor of 1.9 using IL-4 (p < 0.001 and 4.1 using IL-2 (p < 0.001. For donor-versus-unrelated culture responses, no significant increase was observed using IL-4, and a mean response inhibition of 20% was observed using IL-2 (p < 0.001. Neither of the cytokines produced a significant difference in the unrelated control versus recipient cell responses. CONCLUSION: IL-4 supplementation was the best for increasing the mixed lymphocyte culture sensitivity. However, IL-4 also increased autologous responses, albeit less

  6. Optimisation of the CT h4S bioassay for detection of human interleukin-4 secreted by mononuclear cells stimulated by phytohaemaglutinin or by human leukocyte antigen mismatched mixed lymphocyte culture

    DEFF Research Database (Denmark)

    Petersen, Søren Lykke; Russell, Charlotte Astrid; Bendtzen, Klaus

    2002-01-01

    Limiting dilution analysis has been used in the context of allogeneic bone marrow transplantation to determine anti-recipient interleukin-2 (IL-2) producing helper T lymphocyte precursor (HTLp) frequencies, which in several studies have been predictive of graft-versus-host disease (GVHD). Recently...... of IL-4 detection was not due to high amounts of soluble IL-4 receptor. With the use of 1x10(6) responder cells/well in HLA-mismatched MLC, we found limited IL-4 accumulation still increasing at day 12. We conclude that the CT.h4S bioassay is a reliable and specific method for quantification of IL-4...... accumulation in cultures of human MNC. The difference in optimal timing for IL-2 (day 3) and IL-4 (>/=day 12) detection and evidence of very low IL-4 producing HTLp frequencies makes the relevance of a combined IL-2/IL-4 HTLp assay questionable....

  7. Loci controlling lymphocyte production of interferon c after alloantigen stimulation in vitro and their co-localization with genes controlling lymphocyte infiltration of tumors and tumor susceptibility.

    Science.gov (United States)

    Lipoldová, Marie; Havelková, Helena; Badalova, Jana; Vojtísková, Jarmila; Quan, Lei; Krulova, Magdaléna; Sohrabi, Yahya; Stassen, Alphons P; Demant, Peter

    2010-02-01

    Low infiltration of lymphocytes into cancers is associated with poor prognosis, but the reasons why some patients exhibit a low and others a high infiltration of tumors are unknown. Previously we mapped four loci (Lynf1–Lynf4) controlling lymphocyte infiltration of mouse lung tumors. These loci do not encode any of the molecules that are involved in traffic of lymphocytes. Here we report a genetic relationship between these loci and the control of production of IFNγ in allogeneic mixed lymphocyte cultures (MLC). We found that IFNγ production by lymphocytes of O20/A mice is lower than by lymphocytes of OcB-9/Dem mice (both H2pz) stimulated in MLC by irradiated splenocytes of C57BL/10SnPh (H2b) or BALB/ cHeA (H2d) mice, or by ConA. IFNγ production in MLCs of individual (O20 9 OcB-9)F2mice stimulated by irradiated C57BL/10 splenocytes and genotyped for microsatellite markers revealed four IFNγ-controlling loci (Cypr4-Cypr7), each of which is closely linked with one of the four Lynf loci and with a cluster of susceptibility genes for different tumors. This suggests that inherited differences in certain lymphocyte responses may modify their propensity to infiltrate tumors and their capacity to affect tumor growth.

  8. Use of lymphocyte cultures for BrdU replication banding patterns in anuran species (Amphibia

    Directory of Open Access Journals (Sweden)

    Kasahara Sanae

    1998-01-01

    Full Text Available We describe the standardization of lymphocyte culture procedures in order to improve cytological preparations of anuran species. This methodology permits the use of 5-bromodeoxyuridine (BrdU treatment to obtain replication banding patterns in the chromosomes of these species.

  9. Flow microfluorometric DNA content measurements of tissue culture cells and peripheral lymphocytes

    Energy Technology Data Exchange (ETDEWEB)

    Cram, L.S. (Los Alamos Scientific Lab., NM); Lehman, J.M.

    1977-01-01

    The difference in DNA content of peripheral lymphocytes from normal males, normal females, and an individual with a 48 (xxxy) chromosome constitution was determined by rapid flow microfluorometric techniques. A similar comparison was performed using tissue culture fibroblasts derived from an individual with a 49 (xxxxy) chromosome constitution and WI-38 cells as a normal control. Less than 60 min were required to isolate the lymphocytes, to stain the cells fluorescently, and to measure the increased DNA content. The measured increase in DNA content is consistent with chromosome DNA analyses and chromosome length measurements.

  10. Induction of DNA repair synthesis by ultraviolet radiation and methylmethanesulphonate in cultured mouse lymphocytes

    Energy Technology Data Exchange (ETDEWEB)

    Bianchi, V.; Zantedeschi, A.; Levis, A.G. (Padua Univ. (Italy). Ist. di Zoologia e Anatomia Comparata); Ronchese, F. (Inst. of Pathological Anatomy, Padua (Italy))

    1983-02-01

    The induction of DNA repair synthesis by UV-radiation and methylmethanesulphonate (MMS) was studied in mouse lymphocytes and leukemic cells by means of autoradiography and scintillation counting, after labelling in vitro with tritiated thymidine ((/sup 3/H)dThd). Repair stimulation was detected by both procedures in LSTRA AND YC8 leukemic cell lines as well as in primary fibroblasts of BALB/c and BALB/Mo mice. No stimulation was observed in primary cultures of lymphocytes from the spleen, thymus and lymph-nodes of the same mice. In primary lymphocytes neither stimulation with concanavalin A (Con A) nor pre-incubation with 5-bromodeoxyuridine (BUdR) were effective in making evident DNA repair. The data put into question the reliability of the repair test for the prediction of carcinogenic potential of chemicals.

  11. Phosphorylated Myosin Light Chain 2 (p-MLC2) as a Molecular Marker of Antemortem Coronary Artery Spasm

    Science.gov (United States)

    Li, Liliang; Li, Yuhua; Lin, Junyi; Jiang, Jieqing; He, Meng; Sun, Daming; Zhao, Ziqin; Shen, Yiwen; Xue, Aimin

    2016-01-01

    Background It is not uncommon that only mild coronary artery stenosis is grossly revealed after a system autopsy. While coronary artery spasm (CAS) is the suspected mechanism of these deaths, no specific biomarker has been identified to suggest antemortem CAS. Material/Methods To evaluate the potential of using phosphorylated myosin light chain 2 (p-MLC2) as a diagnostic marker of antemortem CAS, human vascular smooth muscle cells (VSMCs) were cultured and treated with common vasoconstrictors, including prostaglandins F2α (PGF2α), acetylcholine (ACh), and 5-hydroxy tryptamine (5-HT). The p-MLC2 level was examined in the cultured cells using Western blot analysis and in a rat model of spasm provocation tests using immunohistochemistry (IHC). Effects of increased p-MLC2 level on VSMCs contractile activities were assessed in vitro using confocal immunofluorescence assay. Four fatal cases with known antemortem CAS were collected and subject to p-MLC2 detection. Results The p-MLC2 was significantly increased in VSMCs after treatments with vasoconstrictors and in the spasm provocation tests. Myofilament was well-organized and densely stained in VSMCs with high p-MLC2 level, but disarrayed in VSMCs with low p-MLC2 level. Three of the 4 autopsied cases showed strongly positive staining of p-MLC2 at the stenosed coronary segment and the adjacent interstitial small arteries. The fourth case was autopsied at the 6th day after death and showed negative-to-mild positive staining of p-MLC2. Conclusions p-MLC2 might be a useful marker for diagnosis of antemortem CAS. Autopsy should be performed as soon as possible to collect coronary arteries for detection of p-MLC2. PMID:27643564

  12. Isolation and Purification of an Early Pregnancy Factor–Like Molecule from Culture Supernatants Obtained from Lymphocytes of Pregnant Women

    OpenAIRE

    1998-01-01

    Purpose:Our purpose was to determine whether lymphocytes synthesize proteins during pregnancy, to observe whether one of the proteins synthesized has early pregnancy factor (EPF)–like activity and to isolate and purify this molecule from culture supernatants obtained from stimulated lymphocytes of pregnant women.

  13. Anti-HIV-1 activity of propolis in CD4(+) lymphocyte and microglial cell cultures.

    Science.gov (United States)

    Gekker, Genya; Hu, Shuxian; Spivak, Marla; Lokensgard, James R; Peterson, Phillip K

    2005-11-14

    An urgent need for additional agents to treat human immunodeficiency virus type 1 (HIV-1) infection led us to assess the anti-HIV-1 activity of the natural product propolis in CD4(+) lymphocytes and microglial cell cultures. Propolis inhibited viral expression in a concentration-dependent manner (maximal suppression of 85 and 98% was observed at 66.6 microg/ml propolis in CD4(+) and microglial cell cultures, respectively). Similar anti-HIV-1 activity was observed with propolis samples from several geographic regions. The mechanism of propolis antiviral property in CD4(+) lymphocytes appeared to involve, in part, inhibition of viral entry. While propolis had an additive antiviral effect on the reverse transcriptase inhibitor zidovudine, it had no noticeable effect on the protease inhibitor indinavir. The results of this in vitro study support the need for clinical trials of propolis or one or more of its components in the treatment of HIV-1 infection.

  14. [Genetic regulation of T-lymphocyte responsiveness to PHA is independent of culture conditions (author's transl)].

    Science.gov (United States)

    Stiffel, C; Liacopoulos-Briot, M; Decreusefond, C; Lambert, F

    1979-01-01

    A maximal interline separation has been obtained after 10 consecutive generations of selective breeding for the character "quantitative in vitro response of lymph node lymphocytes to the mitogenic effect of phytohaemagglutinin". At the selection limit the difference between high and low responder lines was about 20-fold. A similar interline separation has been demonstrated for the T-mitogen effect of concanavalin A. The identical response to PPD (purified protein derivative of tuberculin), a B mitogen, proved that the genetic selection has only modified the potentialities of T lymphocytes. During the selective breeding, responsiveness to PHA stimulation has been always measured under identical culture conditions. To demonstrate that the interline difference in responsiveness was due essentially to genetic factors independent of environmental effects, a systematic study of various culture conditions has been undertaken. The optimal stimulation was found after two days of culture for high line cells and after three days for low line cells. The difference between maximal responses was only slightly lower than that obtained after a two-day culture as used for the selection test. Increase in cell concentrations produced higher thymidine incorporation. In the two lines, a linear correlation was established between the cell concentration and the response produced. The maximal response given by the highest number of low line lymphocytes was equivalent to that given by a number, 11-fold smaller, of high line cells. Within certain limits, changes in the amount of tritiated thymidine added to the culture did not affect the interline separation. With a thymidine of high specific activity, a sub-evaluation of uptake by high line cells decreased the interline difference. Results in mixed culture of lymph node cells from high and low lines indicated that the low response was not due to the release of inhibiting factors or to the presence of suppressive cells in low responder mice

  15. Differential Micronuclei Induction in Human Lymphocyte Cultures by Imidacloprid in the Presence of Potassium Nitrate

    Directory of Open Access Journals (Sweden)

    Polychronis Stivaktakis

    2010-01-01

    Full Text Available Humans are exposed to pesticides as a consequence of their application in farming or their persistence in a variety of media, including food, water, air, soil, plants, animals, and smoke. The interaction of pesticides with environmental factors may result in the alteration of their physicochemical properties. Square wave cathodic stripping voltammetry (SW-CSV, a technique that simulates electrodynamically the cellular membrane, is used to investigate whether the presence of potassium nitrate (KNO3 in the culture medium interferes with the genotoxic behavior of imidacloprid. The cytokinesis block micronuclei (CBMN method is used to evaluate imidacloprid's genotoxicity in the absence or presence of KNO3 in the culture medium and, as a consequence, its adsorption by lymphocytes. Comparing micronuclei (MN frequencies in control and imidacloprid-treated blood cell cultures, statistically significant differences were not detected. KNO3 did not induce MN frequencies compared to control. Statistically significant differences in MN frequencies were observed when blood cell cultures were treated with imidacloprid in the presence of increasing concentrations of KNO3. SW-CSV revealed that by increasing KNO3 molarity, imidacloprid's concentration in the culture medium decreased in parallel. This finding indicates that imidacloprid is adsorbed by cellular membranes. The present study suggests a novel role of a harmless environmental factor, such as KNO3, on the genotoxic behavior of a pesticide, such as imidacloprid. KNO3 rendered imidacloprid permeable to lymphocytes, resulting in elevated MN frequencies.

  16. The cytogenetic effects of black tea and green tea on cultured human lymphocytes

    Directory of Open Access Journals (Sweden)

    Halil Erhan Eroğlu

    2011-12-01

    Full Text Available In this study, the cytogenetic effects of black tea and green tea were determined in cultured peripheral blood lymphocytes. Results showed that black tea and green tea induced the mitotic and replication indexes and decreased micronuclei. But these data were not statistically significant for green tea. The effects of black tea on the micronucleus formation and mitotic index were statistically significant. The decrease in micronucleus counts indicated that black tea and green tea had considerable anticlastogenic and antigenotoxic effects as observed in vitro in human lymphocytes. Thus, it could be concluded that tea polyphenols protected the normal cells from genotoxic or carcinogenic agents, which indicated the therapeutic and antioxidative role of catechins, flavonoids or other tea compounds.

  17. Activation of secretion and surface alteration of cytolytic T-lymphocytes interacting with target cells.

    Science.gov (United States)

    Bykovskaya, S N; Shevelev, A A; Kupriyanova, T A

    1988-01-01

    Cells obtained in mixed lymphocyte culture (MLC) and memory cells adsorbed on the surface of target cells (TC) were examined using scanning and transmission electron microscopy depending on the time of interaction with TC. Three types of lymphocytes were revealed: type I - cells of spherical shape with a smooth surface or an insignificant amount of microvilli; predominantly small and medium-sized lymphocytes contacting TC with non significant involvement of their surface or by several microvilli; type II - oval or round-shaped lymphocytes evenly covered with microvilli with considerably enlarged region of contact; type III cells - predominantly large lymphocytes and lymphoblasts flattened (spread) on TC, with multiple microvilli, ridge-like projections, and ruffles on their surface. TEM revealed activation of the secretory apparatus in the cytoplasm of such lymphocytes. With increased time of interaction, type III cells increase in number (from 8.6% after 10 min to 90.2% after 60 min of incubation). Memory cells show no morphologic signs of secretion in correlation with the absence of lysis of TC on which they are adsorbed. The surface of the lymphocytes adsorbed on the substrate with poly-L-lysin is not noticeably altered. It is suggested that 3 morphological types of lymphocytes correspond to 3 stages of secretion activation. Lymphocyte contact with TC surface is evidently a specific stimulus for activating secretory apparatus of CTL. SEM can be used for quantitation of activated lymphocytes.

  18. Genotoxic and antigenotoxic effects of Fucus vesiculosus extract on cultured human lymphocytes using the chromosome aberration and Comet assays

    Directory of Open Access Journals (Sweden)

    Cleide Leite-Silva

    2007-01-01

    Full Text Available The brown seaweed Fucus vesiculosus (Fucales, Fucaceae was screened for its protective activity using doxorubicin-induced DNA damage in human lymphocytes. In this study, we assessed the genotoxic and antigenotoxic potential of three different concentrations (0.25, 0.5 and 1.0 mg mL-1 of F. vesiculosus aqueous extract using the chromosome aberration and Comet assays. Treatment of human lymphocyte cultures with 0.25, 0.5 and 1.0 mg mL-1 F. vesiculosus aqueous extract had no effect on the chromosome aberration frequency or on the extent of DNA damage detected by the Comet assay. The antigenotoxic effects of the extract were tested in human lymphocyte cultures treated with 15 µg mL-1 of doxorubicin, either alone or combined with the different concentrations of the extract, which was added to the cultures before, simultaneously with or after the doxorubicin. Only when lymphocytes were pre-treated with extract there was a reduction in doxorubicin-induced chromosome aberrations and DNA damage as detected by the Comet assay. These results demonstrate that F. vesiculosus aqueous extract is not genotoxic in cultured human lymphocytes and indicate that when added to lymphocyte cultures before doxorubicin it has antigenotoxic activity against doxorubicin-induced DNA damage.

  19. Protective effects of hesperidin against genotoxicity induced by {sup 99m}Tc-MIBI in human cultured lymphocyte cells

    Energy Technology Data Exchange (ETDEWEB)

    Hosseinimehr, Seyed Jalal [Department of Radiopharmacy, Faculty of Pharmacy and Traditional and Complementary Medicine Research Center, Mazandaran University of Medical Sciences, Sari (Iran, Islamic Republic of)], E-mail: sjhosseinim@yahoo.com; Ahmadi, Amirhossein [Department of Radiopharmacy, Faculty of Pharmacy and Traditional and Complementary Medicine Research Center, Mazandaran University of Medical Sciences, Sari (Iran, Islamic Republic of); Beiki, Davood [Research Institute for Nuclear Medicine, Tehran University of Medical Sciences, Tehran (Iran, Islamic Republic of); Habibi, Emran [Department of Radiopharmacy, Faculty of Pharmacy and Traditional and Complementary Medicine Research Center, Mazandaran University of Medical Sciences, Sari (Iran, Islamic Republic of); Mahmoudzadeh, Aziz [Laboratory of Cytogenetics, Novin Radiation Institute, Tehran (Iran, Islamic Republic of)

    2009-10-15

    Introduction: Radiopharmaceuticals have been widely used as nuclear tracers for myocardial perfusion imaging. The purpose of this study was to investigate the radioprotective effects of hesperidin as a flavonoid which protects against the genotoxic effects of {sup 99m}Tc-MIBI in human cultured lymphocytes. Methods: Whole blood samples from human volunteers were incubated with hesperidin at doses of 10, 50 and 100 {mu}mol. After 1 h of incubation, the lymphocytes were incubated with {sup 99m}Tc-MIBI (200 {mu}Ci/2 ml) for 3 h. The lymphocyte cultures were then mitogenically stimulated to allow for evaluation of the number of micronuclei in cytokinesis-blocked binucleated cells. Results: Incubation of lymphocytes with {sup 99m}Tc-MIBI at this high dose induces additional genotoxicity and shown by increases in micronuclei frequency in human lymphocytes. Hesperidin at these doses significantly reduced the micronuclei frequency in cultured lymphocytes. The maximum protective effect and greatest decrease in micronuclei frequency occurred when cultures were incubated with a 100-{mu}mol dose of 65% hesperidin. Conclusion: This study has important implications for patients undergoing nuclear medicine procedures. The results indicate a protective role for hesperidin against the genetic damage and side effects induced by radiopharmaceutical administration.

  20. Assessment of genotoxicity of Lannate-90® and its plant and animal metabolites in human lymphocyte cultures.

    Science.gov (United States)

    Valencia-Quintana, Rafael; Gómez-Arroyo, Sandra; Sánchez-Alarcón, Juana; Milić, Mirta; Olivares, José Luis Gómez; Waliszewski, Stefan M; Cortés-Eslava, Josefina; Villalobos-Pietrini, Rafael; Calderón-Segura, María Elena

    2016-06-01

    This study evaluated direct and metabolic genotoxic effects caused by Lannate-90®, a methomyl-based formulation (90 % active ingredient), in human lymphocyte cultures using sister chromatid exchange assay (SCE). Two processes were used for the plant promutagens evaluation: in vivo activation, applying the insecticide systemically in plants for 4 h and subsequently adding plant metabolites containing extracts to lymphocyte cultures; and in vitro activation, where the insecticide was incubated with Vicia faba S10 mix plus human lymphocyte culture. Direct treatment with the insecticide significantly increased SCE frequency in human lymphocytes (250-750 mgL-1), with cellular death observed at 1000 mgL-1 concentration. Using the extracts of Vicia faba treated with Lannate-90® to treat human lymphocytes, a dose-response relationship was observed. In lymphocyte cultures treated directly with the insecticide for 2 h, a negative response was obtained. When S10 mix was added, SCE frequency did not change significantly. Meanwhile, a mixture of S9 mammalian metabolic mix and Lannate-90® increased the SCE frequency, with an observed concentration-dependent response. Although Lannate-90® induced cellular death at the highest concentrations, it did not cause a delay in cell proliferation in any of the treatments, confirming its genotoxic action. This study is one of the first to evaluate and compare the direct effect of Lannate-90® in two bioassays, animal and vegetal, and the effect of plant and animal metabolism on its genotoxic potential.

  1. T Lymphocyte Activation Threshold and Membrane Reorganization Perturbations in Unique Culture Model

    Science.gov (United States)

    Adams, C. L.; Sams, C. F.

    2000-01-01

    Quantitative activation thresholds and cellular membrane reorganization are mechanisms by which resting T cells modulate their response to activating stimuli. Here we demonstrate perturbations of these cellular processes in a unique culture system that non-invasively inhibits T lymphocyte activation. During clinorotation, the T cell activation threshold is increased 5-fold. This increased threshold involves a mechanism independent of TCR triggering. Recruitment of lipid rafts to the activation site is impaired during clinorotation but does occur with increased stimulation. This study describes a situation in which an individual cell senses a change in its physical environment and alters its cell biological behavior.

  2. Nuclear anomalies, chromosomal aberrations and proliferation rates in cultured lymphocytes of head and neck cancer patients.

    Science.gov (United States)

    George, Alex; Dey, Rupraj; Bhuria, Vikas; Banerjee, Shouvik; Ethirajan, Sivakumar; Siluvaimuthu, Ashok; Saraswathy, Radha

    2014-01-01

    Head and neck cancers (HNC) are extremely complex disease types and it is likely that chromosomal instability is involved in the genetic mechanisms of its genesis. However, there is little information regarding the background levels of chromosome instability in these patients. In this pilot study, we examined spontaneous chromosome instability in short-term lymphocyte cultures (72 hours) from 72 study subjects - 36 newly diagnosed HNC squamous cell carcinoma patients and 36 healthy ethnic controls. We estimated chromosome instability (CIN) using chromosomal aberration (CA) analysis and nuclear level anomalies using the Cytokinesis Block Micronucleus Cytome Assay (CBMN Cyt Assay). The proliferation rates in cultures of peripheral blood lymphocytes (PBL) were assessed by calculating the Cytokinesis Block Proliferation Index (CBPI). Our results showed a significantly higher mean level of spontaneous chromosome type aberrations (CSAs), chromatid type aberration (CTAs) dicentric chromosomes (DIC) and chromosome aneuploidy (CANEUP) in patients (CSAs, 0.0294±0.0038; CTAs, 0.0925±0.0060; DICs, 0.0213±0.0028; and CANEUPs, 0.0308±0.0035) compared to controls (CSAs, 0.0005±0.0003; CTAs, 0.0058±0.0015; DICs, 0.0005±0.0003; and CANEUPs, 0.0052±0.0013) where pnuclear anomalies showed significantly higher mean level of micronuclei (MNi), nucleoplasmic bridges (NPBs) and nuclear buds (NBUDs) among cases (MNi, 0.01867±0.00108; NPBs, 0.01561±0.00234; NBUDs, 0.00658±0.00068) compared with controls (MNi, 0.00027±0.00009; NPBs, 0.00002±0.00002; NBUDs, 0.00011±0.00007).The evaluation of CBPI supported genomic instability in the peripheral blood lymphocytes showing a significantly lower proliferation rate in HNC patients (1.525±0.005552) compared to healthy subjects (1.686±0.009520 ) (pproliferation in the cultured peripheral lymphocytes of solid tumors could be biomarkers to predict malignancy in early stages.

  3. 糖酵解抑制剂3-溴丙酮酸对异基因小鼠混合培养脾细胞增殖的影响及其机制初步研究%Effect of glycolytic inhibitor 3-BrPA on the proliferation and apoptosis of mouse splenic lymphocytes in mixed lymphocytes culture

    Institute of Scientific and Technical Information of China (English)

    周睿卿; 王静; 邱大发; 牛晓敏; 郭子文; 何慧清; 许晓军

    2014-01-01

    Objective To study the effect of glycolytic inhibitor 3-Bromopyruvate (3-BrPA) on the proliferation and apoptosis of mouse spleen lymphocytes and explore its mechanism.Methods An one-way mixed lymphocyte culture (MLC) system was established,including BALB/c mouse spleen cells (H-2d) as stimulator and C57BL/6 mouse spleen cells (H-2b) as responder.With treatment of 3-BrPA at different concentrations (0-200 μmol/L),lymphocyte proliferation capacity was detected by the CCK-8 method,the expression of CD3,CD4,and CD8 by flow cytometry,and the concentrations of cytokine interleukin (IL)-4 and interferon (IFN)-γ in the supernatant by ELISA.Results At a middle or high dose (over 20 μmol/L),3-BrPA displayed a dose-dependent inhibitory effect on lymphocyte proliferation in the MLC system.The 50% inhibitory concentration (IC50) were 48.6,41.2,and 41.9 μmol/L after 24,36,and 48 h culture,respectively.With treatment of 50 μmol/L 3-BrPA,the IFN-γ level [(164.25±20.14) ng/L] was significantly lower,compared with control [(277.61 ± 18.46) ng/L].The IL-4 level [(31.06 ± 6.06)ng/L] was significantly higher,compared with control [(28.64 ± 3.97) ng/L].Consequently,the IFN-γ/IL-4 ratio decreased significantly.Conclusions These results indicate that 3-BrPA had a significant inhibitory effect on the proliferation of mouse spleen lymphocytes cultured in MLC system,accompanied with the Th2-biased secretion of cytokines.%目的 研究糖酵解抑制剂3-溴丙酮酸(3-BrPA)对单向混合培养的异基因小鼠脾细胞增殖的影响及其机制.方法 以BALB/c(H-2d)小鼠脾细胞为刺激细胞,以C57BL/6(H-2b)小鼠脾细胞为反应细胞,建立单向混合淋巴细胞培养(MLC)体系,给予不同浓度3-BrPA(0~200 μmol/L)处理后,采用CCK-8法检测淋巴细胞增殖活性,流式细胞术检测MLC体系中反应细胞CD3、CD4、CD8等表面标志表达,ELISA法检测MLC体系上清中IL-4、IFN-γ等细胞因子水平.结果 CCK-8检测结果显示,高浓度(20

  4. Cytogenetic study of Ascaris trypsin inhibitor in cultured human lymphocytes with metabolic activation

    Indian Academy of Sciences (India)

    Joanna Blaszkowska; Wanda Bratkowska; Dobroslawa Lopaczynska; Tomasz Ferenc

    2009-04-01

    The trypsin inhibitor (ATI) isolated from gastrointestinal nematode Ascaris suum was tested in vitro for induction of chromosome aberrations and sister chromatid exchanges (SCE). Genotoxicity assessment of purified ATI was carried out on metaphase plates received from peripheral blood lymphocyte macroculture (48 h test of structural chromosome aberrations and 72 h test of SCE) with exogenous metabolic activation. ATI was tested in dose of 25, 50 and 100 μg per ml of culture. Kinetics of cell divisions were determined by the replication index (RI). The mitotic index (MI) was expressed as a number of metaphases per 1000 nuclei analysed. Analysis of chromosome aberrations showed that higher doses of ATI (50 and 100 /ml) significantly increased the frequency of chromosome aberrations (mainly of chromatid gaps and breaks) compared to the negative control. All concentrations of ATI caused a statistically significant reduction in the MI and RI. In comparison with the negative control, a significant increase in the SCE frequency was observed in all applied doses of ATI. Thus, in the presence of S9 activation, the Ascaris trypsin inhibitor showed potential clastogenic activity and inhibition of the dynamics of lymphocyte divisions.

  5. Use of an adaptable cell culture kit for performing lymphocyte and monocyte cell cultures in microgravity

    Science.gov (United States)

    Hatton, J. P.; Lewis, M. L.; Roquefeuil, S. B.; Chaput, D.; Cazenave, J. P.; Schmitt, D. A.

    1998-01-01

    The results of experiments performed in recent years on board facilities such as the Space Shuttle/Spacelab have demonstrated that many cell systems, ranging from simple bacteria to mammalian cells, are sensitive to the microgravity environment, suggesting gravity affects fundamental cellular processes. However, performing well-controlled experiments aboard spacecraft offers unique challenges to the cell biologist. Although systems such as the European 'Biorack' provide generic experiment facilities including an incubator, on-board 1-g reference centrifuge, and contained area for manipulations, the experimenter must still establish a system for performing cell culture experiments that is compatible with the constraints of spaceflight. Two different cell culture kits developed by the French Space Agency, CNES, were recently used to perform a series of experiments during four flights of the 'Biorack' facility aboard the Space Shuttle. The first unit, Generic Cell Activation Kit 1 (GCAK-1), contains six separate culture units per cassette, each consisting of a culture chamber, activator chamber, filtration system (permitting separation of cells from supernatant in-flight), injection port, and supernatant collection chamber. The second unit (GCAK-2) also contains six separate culture units, including a culture, activator, and fixation chambers. Both hardware units permit relatively complex cell culture manipulations without extensive use of spacecraft resources (crew time, volume, mass, power), or the need for excessive safety measures. Possible operations include stimulation of cultures with activators, separation of cells from supernatant, fixation/lysis, manipulation of radiolabelled reagents, and medium exchange. Investigations performed aboard the Space Shuttle in six different experiments used Jurkat, purified T-cells or U937 cells, the results of which are reported separately. We report here the behaviour of Jurkat and U937 cells in the GCAK hardware in ground

  6. Post-thaw non-cultured and post-thaw cultured equine cord blood mesenchymal stromal cells equally suppress lymphocyte proliferation in vitro.

    Directory of Open Access Journals (Sweden)

    Lynn B Williams

    Full Text Available Multipotent mesenchymal stromal cells (MSC are receiving increased attention for their non-progenitor immunomodulatory potential. Cryopreservation is commonly used for long-term storage of MSC. Post-thaw MSC proliferation is associated with a lag-phase in vitro. How this lag-phase affect MSC immunomodulatory properties is unknown. We hypothesized that in vitro there is no difference in lymphocyte suppression potential between quick-thawed cryopreserved equine cord blood (CB MSC immediately included in mixed lymphocyte reaction (MLR and same MSC allowed post-thaw culture time prior to inclusion in MLR. Cryopreserved CB-MSC from five unrelated foals were compared using two-way MLR. For each of the five unrelated MSC cultures, paired MLR assays of MSC allowed five days of post-thaw culture and MSC included in MLR assay immediately post-thawing were evaluated. We report no difference in the suppression of lymphocyte proliferation by CB-MSC that had undergone post-thaw culture and MSC not cultured post-thaw (p<0.0001. Also, there was no inter-donor variability between the lymphocyte suppressive properties of MSC harvested from the five different donors (p = 0.13. These findings suggest that cryopreserved CB-MSC may have clinical utility immediately upon thawing. One implication hereof is the possibility of using cryopreserved CB-MSC at third party locations without the need for cell culture equipment or competencies.

  7. An eigenvalue study of the MLC circuit

    DEFF Research Database (Denmark)

    Lindberg, Erik; Murali, K.

    1998-01-01

    The MLC (Murali-Lakshmanan-Chua) circuit is the simplest non-autonomous chaotic circuit. Insight in the behaviour of the circuit is obtained by means of a study of the eigenvalues of the linearized Jacobian of the nonlinear differential equations. The trajectories of the eigenvalues as functions...

  8. Intra HLA-D/DR region recombinant detected by primed lymphocyte typing (PLT)

    DEFF Research Database (Denmark)

    Jakobsen, B K; Kristensen, T; Lamm, L U

    1983-01-01

    The chromosome 6 markers, HLA-ABC, D, DR, MT, properdin factor Bf, and complement factors 2 (C2) and 5 (C4), were studied in three families, each of which included two HLA identical siblings, one or both of whom were known to be HLA-B: GLO recombinants. The families were also typed with primed...... lymphocyte typing (PLT) for HLA-D/DR region associated DP antigens. None of these studies gave evidence that the recombinations had occurred within the HLA region. Mixed leucocyte culture (MLC) tests within the families showed no detectable stimulation between the HLA identical siblings in two...

  9. Single-Pass, Closed-System Rapid Expansion of Lymphocyte Cultures for Adoptive Cell Therapy

    Science.gov (United States)

    Klapper, Jacob A.; Thomasian, Armen A.; Smith, Douglas M.; Gorgas, Gayle C.; Wunderlich, John R.; Smith, Franz O.; Hampson, Brian S.; Rosenberg, Steven A.; Dudley, Mark E.

    2009-01-01

    Adoptive cell therapy (ACT) for metastatic melanoma involves the ex vivo expansion and re-infusion of tumor infiltrating lymphocytes (TIL) obtained from resected specimens. With an overall objective response rate of fifty-six percent, this T-cell immunotherapy provides an appealing alternative to other therapies, including conventional therapies with lower response rates. However, there are significant regulatory and logistical concerns associated with the ex vivo activation and large scale expansion of these cells. The best current practice uses a rapid expansion protocol (REP) consisting of an ex vivo process that occurs in tissue culture flasks (T-flasks) and gas-permeable bags, utilizes OKT3 (anti-CD3 monoclonal antibody), recombinant human interleukin-2, and irradiated peripheral blood mononuclear cells to initiate rapid lymphocyte growth. A major limitation to the widespread delivery of therapy to large numbers of melanoma patients is the open system in which a REP is initiated. To address this problem, we have investigated the initiation, expansion and harvest at clinical scale of TIL in a closed-system continuous perfusion bioreactor. Each cell product met all safety criteria for patient treatment and by head-to-head comparison had a similar potency and phenotype as cells grown in control T-flasks and gas-permeable bags. However, the currently available bioreactor cassettes were limited in the total cell numbers that could be generated. This bioreactor may simplify the process of the rapid expansion of TIL under stringent regulatory conditions thereby enabling other institutions to pursue this form of ACT. PMID:19389403

  10. Improved renal allograft survival using the mixed lymphocyte culture for selection of nonidentical living related donors.

    Science.gov (United States)

    Riggio, R R; Saal, S D; Katz, E B; Tapia, L; White, R; Chami, J; Sheigh, J S; Sullivan, J F; Stenzel, K H; Stubenbord, W T; Whitsell, J C; Rubin, A L

    1975-01-01

    Our results concur with earlier published work, by other groups, showing that LRD-recipient pairs with low MLC stimulation usually have better and more prolonged graft success than do those with higher stimulation. Specific HL-A compatibilities or incompatibilities did not seem to affect these results, nor did the presence of an increased number of common loci, short of increasing the apparent chromosome compatibility. The presence of pre-transplant cytotoxic antibodies, in patients with a high MI, however, may unfavorably affect the LRD transplant. The overall results of our LRD transplant experience is shown in Figure 1, and superimposed upon Figure 2, is the current extrapolation of data showing MLC stimulation and haplotype success. Thus, it appears that graft survival may be improved and more closely approach the levels seen in a full-house, diplotype match, by using the MLC results in considering patients for transplantation. Not all patients with a high MLC, however, (see table) reject their grafts and it is impossible to predict pre-transplant who will develop specific allograft enhancement. Before the MI becomes a specific criteria for transplant selection, additional studies of patient stimulation in MLC should be done. Suppression of stimulation by donor cells in autologous serum, as compared to the response to unrelated controls, might provide pre-transplant clues to the presence of enhancing factors. Such studies could provide an index that would be more meaningful than the MI in AB sera alone. Since overall results from both our series and from the Transplant Registry continue to indicate better long term graft survival for LRD than for cadaver transplants, and since the evidence suggests that a successful transplant offers a patient a better quality of life, as well as decreased morbidity and mortality compared to concomitant time spent on hemodialysis, continued LRD transplants with high MI is warranted in some circumstances with the patient

  11. DNA repair capacity of cultured human lymphocytes exposed to mutagens measured by the comet assay and array expression analysis.

    Science.gov (United States)

    Bausinger, Julia; Speit, Günter

    2015-11-01

    Repair of mutagen-induced DNA lesions during transportation, storage and cultivation of lymphocytes may have a significant impact on results obtained in human biomonitoring after occupational and environmental exposure of human populations to genotoxic chemicals. Using the comet assay in combination with the repair inhibitor aphidicolin and array gene expression analysis of 92 DNA repair genes, we investigated the repair of DNA lesions induced by methyl methanesulfonate (MMS) and benzo[a]pyrenediolepoxide (BPDE) in phytohaemagglutinin (PHA)-stimulated cultured human lymphocytes in the time segment before replication. The comet assay indicated fast repair of MMS-induced damage during the first hours of cultivation. In contrast, removal of BPDE-induced lesions was slower and significant amounts of damage seem to persist until S-phase. Gene expression analysis revealed that PHA stimulation had a clear effect on gene regulation in lymphocytes already during the first 18h of cultivation. Under the conditions of this study, genotoxic concentrations of MMS did not induce significant changes in gene expression. In contrast, exposure to BPDE led to altered expression of several genes in a time- and concentration-related manner. Of the significantly up-regulated genes, only two genes (XPA and XPC) were directly related to nucleotide excision repair. Our results suggest that PHA stimulation of human lymphocytes influences the expression of DNA repair genes in human lymphocytes. The effect of induced DNA damage on gene expression is comparatively low and depends on the mutagens used. PHA-stimulated lymphocytes repair induced DNA damage before they start to replicate but the repair activity during the first 18h of cultivation is not affected by changes in the expression of DNA repair genes during this period of time.

  12. Dioxidine-induced changes in genome-wide DNA methylation in a culture of peripheral blood lymphocytes.

    Science.gov (United States)

    Smirnikhina, S A; Voronina, E S; Lavrov, A V; Bochkov, N P

    2013-06-01

    We studied the effect of dioxidine on genome-wide methylation in short-term cultures of peripheral blood lymphocytes derived from healthy donors. Methylation was evaluated in lymphocytes before culturing, after 25 h in culture, and 1 h after addition of dioxidine in two concentrations (0.1 and 0.01 mg/ml). The total time in culture was 25 h. The level of methylation was assessed using methyl-sensitive single-cell gel electrophoresis ("comet assay") with additional restriction with HpaII amd MspI. Significant individual differences were found in the levels of methylation in both native cells and in cells treated with dioxidine in both concentrations. Mean group indicators of methylation did not differ before culturing and after 25 h in culture (45.28 and 44.80%, respectively). The mean group rate of methylation increased to 46.14% (p<0.001) after dioxidine treatment in a concentration of 0.01 mg/ml. Dioxidine in 0.1 mg/ml reduced the level of methylation (mean group rate 42.31%; p<0.001).

  13. Tumor infiltrating lymphocytes in melanoma comprise high numbers of T-cell clonotypes that are lost during in vitro culture

    DEFF Research Database (Denmark)

    thor Straten, P; Kirkin, A F; Siim, E

    2000-01-01

    Melanoma is generally accepted as being an antigenic tumor capable of eliciting T-cell responses that, however, in most cases are inadequate to control tumor growth. Tumor-infiltrating lymphocytes (TIL) in melanoma lesions comprise clonotypic T cells, indicating the in situ recognition of melanoma......-associated peptide epitopes. Cultured TIL have been studied in order to unveil characteristics of TIL and the interactions of TIL and melanoma cells. Whether in vitro cultured TIL mirrors the in situ situation has, however, been questioned. In the present study we have taken advantage of T-cell receptor clonotype...

  14. Abnormal humoral immune responses in peripheral blood lymphocyte cultures of bone marrow transplant recipients.

    Science.gov (United States)

    Pahwa, S G; Pahwa, R N; Friedrich, W; O'Reilly, R J; Good, R A

    1982-01-01

    The present study was aimed at investigating recovery of humoral immunity in vitro after bone marrow transplantation in patients with acute leukemia and severe aplastic anemia. Hemolytic plaque assays were utilized to quantitate pokeweed mitogen-stimulated polyclonal immunoglobulin production and sheep erythrocyte antigen-specific antibody responses in cultures of peripheral blood mononuclear cells of 39 patients beginning at 1 month, for variable periods up to a maximum of 4 years after marrow transplantation. Three phases were identified: an early period of primary B cell dysfunction with concomitant immunoregulatory T cell abnormalities--i.e., decreased helper and increased suppressor activities; an intermediate phase in which B cell dysfunction could be attributed in large measure to immunoregulatory T cell abnormalities; and a late phase of normal B and T lymphocyte functions. Patients with graft-versus-host disease differed from those without it in that they often did not manifest increased T cell suppressor activity in the early period, and they were noted to have prolonged and profound B and T cell abnormalities in the chronic phase of their disease. In selected patients, simultaneous assessment of ratios of Leu-2 to Leu-3 antigens on T cells by monoclonal antibodies and of immunoregulatory T cell functions revealed a correlation between the two only late in the post-transplant period. These studies provide an insight into the ontogeny of B cell function in the post-transplant period and indicate that in certain situations phenotypic alterations in T cell subsets cannot reliably be used to predict abnormalities in their function in recipients of marrow transplantation. Images PMID:6211673

  15. DEPTOR-mTOR Signaling Is Critical for Lipid Metabolism and Inflammation Homeostasis of Lymphocytes in Human PBMC Culture

    Directory of Open Access Journals (Sweden)

    Qi-bing Xie

    2017-01-01

    Full Text Available Abnormal immune response of the body against substances and tissues causes autoimmune diseases, such as polymyositis, dermatomyositis, and rheumatoid arthritis. Irregular lipid metabolism and inflammation may be a significant cause of autoimmune diseases. Although much progress has been made, mechanisms of initiation and proceeding of metabolic and inflammatory regulation in autoimmune disease have not been well-defined. And novel markers for the detection and therapy of autoimmune disease are urgent. mTOR signaling is a central regulator of extracellular metabolic and inflammatory processes, while DEP domain-containing mTOR-interacting protein (DEPTOR is a natural inhibitor of mTOR. Here, we report that overexpression of DEPTOR reduces mTORC1 activity in lymphocytes of human peripheral blood mononuclear cells (PBMCs. Combination of DEPTOR overexpression and mTORC2/AKT inhibitors effectively inhibits lipogenesis and inflammation in lymphocytes of PBMC culture. Moreover, DEPTOR knockdown activates mTORC1 and increases lipogenesis and inflammations. Our findings provide a deep insight into the relationship between lipid metabolism and inflammations via DEPTOR-mTOR pathway and imply that DEPTOR-mTOR in lymphocytes of PBMC culture has the potential to be as biomarkers for the detection and therapies of autoimmune diseases.

  16. DEPTOR-mTOR Signaling Is Critical for Lipid Metabolism and Inflammation Homeostasis of Lymphocytes in Human PBMC Culture

    Science.gov (United States)

    Liang, Yan; Yang, Yuan

    2017-01-01

    Abnormal immune response of the body against substances and tissues causes autoimmune diseases, such as polymyositis, dermatomyositis, and rheumatoid arthritis. Irregular lipid metabolism and inflammation may be a significant cause of autoimmune diseases. Although much progress has been made, mechanisms of initiation and proceeding of metabolic and inflammatory regulation in autoimmune disease have not been well-defined. And novel markers for the detection and therapy of autoimmune disease are urgent. mTOR signaling is a central regulator of extracellular metabolic and inflammatory processes, while DEP domain-containing mTOR-interacting protein (DEPTOR) is a natural inhibitor of mTOR. Here, we report that overexpression of DEPTOR reduces mTORC1 activity in lymphocytes of human peripheral blood mononuclear cells (PBMCs). Combination of DEPTOR overexpression and mTORC2/AKT inhibitors effectively inhibits lipogenesis and inflammation in lymphocytes of PBMC culture. Moreover, DEPTOR knockdown activates mTORC1 and increases lipogenesis and inflammations. Our findings provide a deep insight into the relationship between lipid metabolism and inflammations via DEPTOR-mTOR pathway and imply that DEPTOR-mTOR in lymphocytes of PBMC culture has the potential to be as biomarkers for the detection and therapies of autoimmune diseases. PMID:28349073

  17. Radiation-induced chromosomal hot spots at G 1 and G 2 stages of human lymphocytes in culture

    Directory of Open Access Journals (Sweden)

    Murugesan R

    2003-01-01

    Full Text Available Radiation-induced chromosomal break points in cultured lymphocytes of normal healthy individuals as well as of those with certain genetic disorders are reported to be localized at certain specific loci (hot spots. These reports are based on studies carried out in lymphocytes irradiated at G 1 stage. The present study examines whether the location of hot spots and the frequency seen in cells irradiated at G 1 are similar to those irradiated at G 2 stage of the cell cycle and also tests whether cells of patients exhibit hot spots on irradiation.The results showed that the radiation induced chromosomal break points to be similar in those irradiated are G 1 and G 2 stages of the cell cycle and also that cells of patients exhibited chromosomal hot spots.

  18. Genotoxicity and cytotoxicity of copper oxychloride in cultured human lymphocytes using cytogenetic and molecular tests.

    Science.gov (United States)

    Bayram, Suleyman; Genc, Ahmet; Buyukleyla, Mehmet; Rencuzogullari, Eyyup

    2016-10-01

    The genotoxicity of copper oxychloride was investigated in human lymphocytes using chromosome aberration (CA) and micronucleus (MN) tests and the randomly amplified polymorphic DNA-polymerase chain reaction technique. The lymphocytes were treated with 3, 6, and 12 µg/mL of copper oxychloride for 24 and 48 h. Copper oxychloride increased CA and abnormal cells in a dose-dependent manner. The frequency of MN and micronucleated binuclear cells also increased at all concentrations and treatment periods. However, copper oxychloride cytotoxicity, observed through lower mitotic and nuclear division index, was significantly lower only at the higher concentrations (6 and 12 µg/mL). Copper oxychloride increased the polymorphic bands and decreased genomic template stability. In conclusion, in this study it was confirmed that copper oxychloride has genotoxic potential for human lymphocytes in vitro. Additionally, caution is advised for its use as a fungicide, because it may increase the risk of exposure through the food chain.

  19. UV-induced chromosome aberrations, sister-chromatid exchanges and cell survival in cultured lymphocytes from malnourished children

    Energy Technology Data Exchange (ETDEWEB)

    Murthy, P.B.; Rahiman, M.A.; Tulpule, P.G.

    1982-08-01

    Cultured lymphocytes from children with kwashiorkor and from normal children were examined for their susceptibility to ultraviolet (UV)-induced chromosome aberrations, sister-chromatid exchanges and cell survival. Cells from kwashiorkor exhibited increased chromosome aberrations, but not sister-chromatid exchanges, when exposed to higher doses of UV. Furthermore, when cells from these patients were exposed to higher doses of UV, there was a significant reduction in viability. These results indicate that, as compared to normals, cells from kwashiorkor were more sensitive to the lethal effects of UV.

  20. In vitro culture of skin-homing T lymphocytes from inflammatory skin diseases

    DEFF Research Database (Denmark)

    Bang, Karen; C Mogensen, Søren; Thestrup-Pedersen, Kristian

    2005-01-01

    -chain studies by using reverse transcription-polymerase chain reaction showed that all T lymphocytes had access to the skin compartment. Single-stranded conformational analysis showed clonally expanded T cells numbering between 40 and 60 clones. After approximately 2 months of growth, the mean CD4+ : CD8+ ratio...

  1. Dynamic MLC-QA Based On Portal Dosimetry

    Directory of Open Access Journals (Sweden)

    Sajeev Surendran

    2014-05-01

    Full Text Available Purpose: Intensity modulated radiotherapy using dynamic delivery method requires accurate verification of MLC, its position and speed of motion. These parameter have major impact on dose delivery on patients. For quality assurance (QA procedure requires more time consumed in a radiotherapy department. The main purpose of this study was to investigate the potential use of amorphous silicon based EPID portal dosimetry for dMLC QA Methods and Materials: A varian Clinac_iX with On Board Imager (OBI and Rapid Arc facility ( VMAT equipped with 120 leaf Millennium MLC and with Amorphous Silicon Based EPID (aSi-1000, varian mounted on a Exact Robotic Arm is used. The dMLC QA consists of different dynamic MLC pattern provided by varian for checking positional accuracy, MLC gap, Leaf speed and complex dynamic field. Results and Discussion: Various dMLC tests were done using portal dosimetry. All results are within the tolerance limit. Picket fence test shows that leaf position errors of upto 0.2mm can be detected which are within the tolerance limit. Complex dynamic field were exposed to EPID, which shows the leaf speed and are within the tolerance limit.

  2. Anti-oxidative and anti-genotoxic effects of methanolic extract of Mentha pulegium on human lymphocyte culture.

    Science.gov (United States)

    Alpsoy, Lokman; Sahin, Hilal; Karaman, Seyda

    2011-08-01

    In the present work, methanolic extract of Mentha pulegium from Erzurum, Turkey, was used in order to report the results of anti-oxidant capacity, anti-oxidant activity and anti-genotoxic effects. The total antioxidant capacity and total phenolic content were measured by using CUPRAC, ABTS and Folin-Ciocalteu colorimetric methods. The total phenolic content was higher than the total antioxidant capacity (for the results of both the CUPRAC and ABTS methods) of methanolic extract of M pulegium (ME). Also, we evaluated the anti-oxidant enzyme activity such as superoxide dismutase (SOD) and glutation peroxidase, total glutation (GSH) and malondialdehyde (MDA) in human lymphocyte culture. In CCl₄-treated group, the activity of SOD, glutathione peroxidase (GPx) and GSH decreased significantly and the level of MDA increased significantly. A significant increase in the activity of SOD, GPx and the level of GSH were seen when supplemented with ME to CCl₄-treated group. Furthermore, a significant decrease in the level of MDA was observed when compared with CCl₄ alone treated group. In addition, anti-genotoxic effect of ME was studied by using sister chromatid exchange (SCE) method. As a result, ME has shown anti-genotoxic effect depend on anti-oxidative effect on human lymphocyte culture.

  3. Antimutagenic effect of aqueous extract from Agaricus brasiliensis on culture of human lymphocytes.

    Science.gov (United States)

    Gameiro, Paula H; Nascimento, José S; Rocha, Beatriz H G; Piana, Clause F B; Santos, Raquel A; Takahashi, Catarina S

    2013-02-01

    The mushroom Agaricus brasiliensis (sun mushroom), native from the southeast of Brazil, is well known by its medicinal properties that include effects on diabetes, cholesterol levels, and osteoporosis. The antimutagenic effects of A. brasiliensis has been investigated recently and revealed some controversial results depending on the temperature by which the A. brasiliensis tea is obtained. In the present study, we evaluated the effect of the A. brasiliensis extract prepared in two different temperatures, 4°C and 25°C, on the doxorubicin-induced DNA strand breaks and chromosomal aberrations (CAs) in human lymphocytes. The results demonstrated that A. brasiliensis was able to reduce the DXR-induced DNA damage in both temperatures; however, the CA test was more sensitive to demonstrate a better reduction when the cells were treated with an extract obtained at 25°C. A. brasiliensis extract obtained in different temperatures exhibited antigenotoxic and anticlastogenic effects in human lymphocytes.

  4. Astaxanthin, a Carotenoid, Stimulates Immune Responses by Enhancing IFN-γ and IL-2 Secretion in Primary Cultured Lymphocytes in Vitro and ex Vivo.

    Science.gov (United States)

    Lin, Kuan-Hung; Lin, Kao-Chang; Lu, Wan-Jung; Thomas, Philip-Aloysius; Jayakumar, Thanasekaran; Sheu, Joen-Rong

    2015-12-29

    Astaxanthin, a potent antioxidant carotenoid, plays a major role in modulating the immune response. In this study, we examined the immunomodulatory effects of astaxanthin on cytokine production in primary cultured lymphocytes both in vitro and ex vivo. Direct administration of astaxanthin (70-300 nM) did not produce cytotoxicity in lipopolysaccharide (LPS, 100 µg/ mL)- or concanavalin A (Con A, 10 µg/ mL)-activated lymphocytes, whereas astaxanthin alone at 300 nM induced proliferation of splenic lymphocytes (p astaxanthin, alone or with Con A, had no apparent effect on interferon (INF-γ) and interleukin (IL-2) production in primary cultured lymphocytes, it enhanced LPS-induced INF-γ production. In an ex vivo experiment, oral administration of astaxanthin (0.28, 1.4 and 7 mg/kg/day) for 14 days did not cause alterations in the body or spleen weights of mice and also was not toxic to lymphocyte cells derived from the mice. Moreover, treatment with astaxanthin significantly increased LPS-induced lymphocyte proliferation ex vivo but not Con A-stimulated lymphocyte proliferation ex vivo. Enzyme linked immunosorbent assay (ELISA) analysis revealed that administration of astaxanthin significantly enhanced INF-γ production in response to both LPS and Con A stimulation, whereas IL-2 production increased only in response to Con A stimulation. Also, astaxanthin treatment alone significantly increased IL-2 production in lymphocytes derived from mice, but did not significantly change production of INF-γ. These findings suggest that astaxanthin modulates lymphocytic immune responses in vitro, and that it partly exerts its ex vivo immunomodulatory effects by increasing INF-γ and IL-2 production without inducing cytotoxicity.

  5. An alternative approach to studying the effects of ZnO nanoparticles in cultured human lymphocytes: combining electrochemistry and genotoxicity tests.

    Science.gov (United States)

    Branica, Gina; Mladinić, Marin; Omanović, Dario; Želježić, Davor

    2016-12-01

    Nanoparticle use has increased radically raising concern about possible adverse effects in humans. Zinc oxide nanoparticles (ZnO NPs) are among the most common nanomaterials in consumer and medical products. Several studies indicate problems with their safe use. The aim of our study was to see at which levels ZnO NPs start to produce adverse cytogenetic effects in human lymphocytes as an early attempt toward establishing safety limits for ZnO NP exposure in humans. We assessed the genotoxic effects of low ZnO NP concentrations (1.0, 2.5, 5, and 7.5 μg mL-1) in lymphocyte cultures over 14 days of exposure. We also tested whether low and high-density lymphocytes differed in their ability to accumulate ZnO NPs in these experimental conditions. Primary DNA damage (measured with the alkaline comet assay) increased with nanoparticle concentration in unseparated and high density lymphocytes. The same happened with the fragmentation of TP53 (measured with the comet-FISH). Nanoparticle accumulation was significant only with the two highest concentrations, regardless of lymphocyte density. High-density lymphocytes had significantly more intracellular Zn2+ than light-density ones. Our results suggest that exposure to ZnO NPs in concentrations above 5 μg mL-1 increases cytogenetic damage and intracellular Zn2+ levels in lymphocytes.

  6. Isolation and evaluation of cytogenetic effect of Brahmi saponins on cultured human lymphocytes exposed in vitro.

    Science.gov (United States)

    Kalachaveedu, Mangathayaru; Papacchan, Sunu; Sanyal, Sudip; Koshy, Teena; Telapolu, Srivani

    2015-01-01

    Major saponins of Brahmi (Bacopa monniera, Fam: Scrophulariaceae) - bacosides A and B - were isolated from the total methanol extract and characterised based on melting point, TLC, IR, (1)H NMR and (13)C NMR. They were evaluated for their in vitro cytogenetic effects on human peripheral blood lymphocytes by chromosomal aberration (CA) assay and sister chromatid exchange (SCE) assay. The frequency of chromatid type aberrations and reciprocal interchanges between sister chromatids in the treated cells was scored in comparison to the untreated control. At 30 μg/mL dose, bacoside A showed a statistically significant increase in the frequency of both CA and SCE and bacoside B showed an increase only in SCE. Our report of the genotoxicity of the saponins is significant in view of the reports of anticancer activity of Brahmi extracts.

  7. Monte Carlo study of MLC fields for cobalt therapy machine

    Directory of Open Access Journals (Sweden)

    Komanduri M Ayyangar

    2014-01-01

    Full Text Available An automated Multi-Leaf Collimator (MLC system has been developed as add-on for the cobalt-60 teletherapy machines available in India. The goal of the present computational study is to validate the MLC design using Monte Carlo (MC modeling. The study was based on the Kirloskar-supplied Phoenix model machines that closely match the Atomic Energy of Canada Limited (AECL theratron-80 machine. The MLC is a retrofit attachment to the collimator assembly, with 14 non-divergent leaf pairs of 40 mm thick, 7 mm wide, and 150 mm long tungsten alloy plates with rounded edges and 20 mm tongue and 2 mm groove in each leaf. In the present work, the source and collimator geometry has been investigated in detail to arrive at a model that best represents the measured dosimetric data. The authors have studied in detail the proto-I MLC built for cobalt-60. The MLC field sizes were MC simulated for 2 × 2 cm 2 to 14 × 14 cm 2 square fields as well as irregular fields, and the percent depth dose (PDD and profile data were compared with ROPS† treatment planning system (TPS. In addition, measured profiles using the IMATRIXX system‡ were also compared with the MC simulations. The proto-I MLC can define radiation fields up to 14 × 14 cm΂ within 3 mm accuracy. The maximum measured leakage through the leaf ends in closed condition was 3.4% and interleaf leakage observed was 7.3%. Good agreement between MC results, ROPS and IMATRIXX results has been observed. The investigation also supports the hypothesis that optical and radiation field coincidence exists for the square fields studied with the MLC. Plots of the percent depth dose (PDD data and profile data for clinically significant irregular fields have also been presented. The MC model was also investigated to speed up the calculations to allow calculations of clinically relevant conformal beams.

  8. Chronic Brucellosis Patients Retain Low Frequency of CD4+ T-Lymphocytes Expressing CD25 and CD28 after Escherichia coli LPS Stimulation of PHA-Cultured PBMCs

    Directory of Open Access Journals (Sweden)

    Panagiotis Skendros

    2008-01-01

    Full Text Available Chronic brucellosis patients display a defective Th1 response to PHA. We have previously shown that heat-killed B. abortus (HKBA can downregulate the PHA-induced increase of CD4+/CD25+ and CD14+/CD80+ cells of brucellosis patients. In the present study, we investigate the effect of E. coli LPS, as a potent stimulant of monocytes and autologous T-lymphocytes, on the PHA-cultured PBMCs of the same groups of patients. Thirteen acute brucellosis (AB patients, 22 chronic brucellosis (CB patients, 11 “cured” subjects, and 15 healthy volunteers were studied. The percentage of CD4+/CD25+ and CD4+/CD28+ T-lymphocytes as well as CD14+/CD80+ monocytes were analyzed by flow cytometry after PBMCs culture with PHA plus E. coli LPS. A significant decrease in the percentage of CD4+/CD25+ and CD4+/CD28+ T-lymphocytes was observed in CB compared to AB. In HKBA cultures, compared to E. coli LPS-cultures, there was a significant reduction of CD4+/CD25+ T-lymphocytes in all groups and CD14+/CD80+ in patients groups. We suggest that Brucella can modulate host immune response, leading to T-cell anergy and chronic infection.

  9. Inhibitory effect of mycoplasma-released arginase. Activity in mixed-lymphocyte and tumour cell cultures

    DEFF Research Database (Denmark)

    Claesson, M H; Tscherning, T; Nissen, Mogens Holst

    1990-01-01

    Non-fermenting mycoplasma species deplete culture media for arginine through arginase activity linked to their arginine deiminase pathway, resulting in proliferation arrest and cell death in mycoplasma-contaminated cell cultures. The presence of only 2-3 Mycoplasma (M.) arginini-contaminated T...... inhibition can be reversed by addition of excess arginine to the culture medium. Antisera raised against non-fermenting, but not against fermenting, mycoplasma species block the inhibitory effect of MAE. SDS-PAGE separation of MAE disclosed a broad band at 60 kDa which contained arginase activity when...

  10. Viability of lymphocyte culture, at different times after blood collection, for karyotype analysis

    Directory of Open Access Journals (Sweden)

    Ligia Maria Crubelati Bulla

    2014-04-01

    Full Text Available Introduction:Cytogenetics is the area of genetics that studies chromosomes, including numerical changes, and their relationship to structural imbalances. Among the classical cytogenetics tests, the GTG banding karyotype is the most widely used. The period of culture establishment is a critical step, which can affect the pre-analytical phase of the test.Objective:To evaluate, at different establishment times, culture viability and banding resolution.Material and methods:Collection of 10 ml blood from 10 subjects was carried out for culture analysis. For viability analysis, mitotic index (MI and banding resolution were assessed. Results: The comparative analysis of MI showed significant difference between times. In the assessment of banding resolution, the mean value of the bands was higher at times zero and 24 hour.Discussion:The MI reflects inhibition of cell cycle progression and/or loss of ability to proliferate. When the pair analysis was performed, a difference between zero and 48 hours was observed. The average number of bands analyzed at times zero and 24 hours did not indicate difference in the quantity and quality of the bands when cultures were grown immediately after blood collection or within 24 hours. At the 48th hour after blood collection significant reduction of band resolution was observed.Conclusion:These data highlight the importance of the biological material quality, as viability is lower when the culture is grown after 24 hours, as well as the banding resolution.

  11. MLC1 protein: a likely link between leukodystrophies and brain channelopathies

    Directory of Open Access Journals (Sweden)

    Maria Stefania eBrignone

    2015-04-01

    Full Text Available Megalencephalic leukoencephalopathy with subcortical cysts (MLC disease is a rare inherited, autosomal recessive form of childhood-onset spongiform leukodystrophy characterized by macrocephaly, deterioration of motor functions, epileptic seizures and mental decline. Brain edema, subcortical fluid cysts, myelin and astrocyte vacuolation are the histopathological hallmarks of MLC. Mutations in either the MLC1 gene (>75% of patients or the GlialCAM gene (<20% of patients are responsible for the disease. Recently, the GlialCAM adhesion protein was found essential for the membrane expression and function of the chloride channel ClC-2 indicating MLC disease caused by mutation in GlialCAM as the first channelopathy among leukodystrophies.. These results may explain the phenotypic convergence of ClC-2 KO mice showing brain edema and myelin vacuolation and human MLC pathology, despite mutations in the ClC-2 gene were not found in patients affected by this leukodystrophy. On the contrary, the function of MLC1 protein, which binds GlialCAM, its functional relationship with ClC-2 and the molecular mechanisms underlying MLC1 mutation-induced functional defects are not fully understood yet. The human MLC1 gene encodes a 377-amino acid membrane protein with eight predicted transmembrane domains which shows very low homology with voltage-dependent potassium K+ channel subunits. The high expression of MLC1 in brain astrocytes contacting blood vessels and meninges and brain alterations observed in MLC patients have led to hypothesize a role for MLC1 in the regulation of ion and water homeostasis. Recent studies have shown that MLC1 establishes structural and/or functional interactions with several ion/water channels and transporters and ion channel accessory proteins, and that these interactions are affected by MLC1 mutations causing MLC. Here, we review data on MLC1 functional properties obtained in in vitro and in vivo models and discuss evidence linking the

  12. Real-Time Dynamic MLC Tracking for Intensity Modulated Arc Therapy

    DEFF Research Database (Denmark)

    Falk, Marianne

    -fraction motion management that uses the MLC of the treatment machine to reshape the beam to follow the tumour movements. The 3D MLC tracking algorithm recalculates the planned MLC positions using information about the tumour location from an independent monitoring system and the leaves are adjusted accordingly...

  13. Real-Time Dynamic MLC Tracking for Intensity Modulated Arc Therapy

    DEFF Research Database (Denmark)

    Falk, Marianne

    -fraction motion management that uses the MLC of the treatment machine to reshape the beam to follow the tumour movements. The 3D MLC tracking algorithm recalculates the planned MLC positions using information about the tumour location from an independent monitoring system and the leaves are adjusted accordingly...

  14. Fast regional readout CMOS Image Sensor for dynamic MLC tracking

    Science.gov (United States)

    Zin, H.; Harris, E.; Osmond, J.; Evans, P.

    2014-03-01

    Advanced radiotherapy techniques such as volumetric modulated arc therapy (VMAT) require verification of the complex beam delivery including tracking of multileaf collimators (MLC) and monitoring the dose rate. This work explores the feasibility of a prototype Complementary metal-oxide semiconductor Image Sensor (CIS) for tracking these complex treatments by utilising fast, region of interest (ROI) read out functionality. An automatic edge tracking algorithm was used to locate the MLC leaves edges moving at various speeds (from a moving triangle field shape) and imaged with various sensor frame rates. The CIS demonstrates successful edge detection of the dynamic MLC motion within accuracy of 1.0 mm. This demonstrates the feasibility of the sensor to verify treatment delivery involving dynamic MLC up to ~400 frames per second (equivalent to the linac pulse rate), which is superior to any current techniques such as using electronic portal imaging devices (EPID). CIS provides the basis to an essential real-time verification tool, useful in accessing accurate delivery of complex high energy radiation to the tumour and ultimately to achieve better cure rates for cancer patients.

  15. Monte Carlo study of MLC fields for cobalt therapy machine

    Science.gov (United States)

    Ayyangar, Komanduri M.; Rani, Roopa A.; Kumar, Anil; Reddy, A. R.

    2014-01-01

    An automated Multi-Leaf Collimator (MLC) system has been developed as add-on for the cobalt-60 teletherapy machines available in India. The goal of the present computational study is to validate the MLC design using Monte Carlo (MC) modeling. The study was based on the Kirloskar-supplied Phoenix model machines that closely match the Atomic Energy of Canada Limited (AECL) theratron-80 machine. The MLC is a retrofit attachment to the collimator assembly, with 14 non-divergent leaf pairs of 40 mm thick, 7 mm wide, and 150 mm long tungsten alloy plates with rounded edges and 20 mm tongue and 2 mm groove in each leaf. In the present work, the source and collimator geometry has been investigated in detail to arrive at a model that best represents the measured dosimetric data. The authors have studied in detail the proto-I MLC built for cobalt-60. The MLC field sizes were MC simulated for 2 × 2 cm2 to 14 × 14 cm2 square fields as well as irregular fields, and the percent depth dose (PDD) and profile data were compared with ROPS† treatment planning system (TPS). In addition, measured profiles using the IMATRIXX system‡ were also compared with the MC simulations. The proto-I MLC can define radiation fields up to 14 × 14 cm2 within 3 mm accuracy. The maximum measured leakage through the leaf ends in closed condition was 3.4% and interleaf leakage observed was 7.3%. Good agreement between MC results, ROPS and IMATRIXX results has been observed. The investigation also supports the hypothesis that optical and radiation field coincidence exists for the square fields studied with the MLC. Plots of the percent depth dose (PDD) data and profile data for clinically significant irregular fields have also been presented. The MC model was also investigated to speed up the calculations to allow calculations of clinically relevant conformal beams. †Radiation Oncology Planning System (ROPS) is supplied by Tirumala Jyothi Computer Systems described at https

  16. Evaluating efficiency of coaxial MLC VMAT plan for spine SBRT

    Energy Technology Data Exchange (ETDEWEB)

    Son, Sang Jun; Mun, Jun Ki; Kim, Dae Ho; Yoo, Suk Hyun [Dept. of Radiation Oncology, Seoul National University Hospital, Seoul (Korea, Republic of)

    2014-12-15

    The purpose of the study is to evaluate the efficiency of Coaxial MLC VMAT plan (Using 273° and 350° collimator angle) That the leaf motion direction aligned with axis of OAR (Organ at risk, It means spinal cord or cauda equine in this study.) compare to Universal MLC VMAT plan (using 30° and 330 ° collimator angle) for spine SBRT. The 10 cases of spine SBRT that treated with VMAT planned by Coaxial MLC and Varian TBX were enrolled. Those cases were planned by Eclipse (Ver. 10.0.42, Varian, USA), PRO3 (Progressive Resolution Optimizer 10.0.28) and AAA (Anisotropic Analytic Algorithm Ver. 10.0.28) with coplanar 260 ° arcs and 10MV FFF (Flattening filter free). Each arc has 273° and 350 ° collimator angle, respectively. The Universal MLC VMAT plans are based on existing treatment plans. Those plans have the same parameters of existing treatment plans but collimator angle. To minimize the dose difference that shows up randomly on optimizing, all plans were optimized and calculated twice respectively. The calculation grid is 0.2 cm and all plans were normalized to the target V100%=90%. The indexes of evaluation are V10Gy, D0.03cc, Dmean of OAR (Organ at risk, It means spinal cord or cauda equine in this study.), H.I (Homogeneity index) of the target and total MU. All Coaxial VMAT plans were verified by gamma test with Mapcheck2 (Sun Nuclear Co., USA), Mapphan (Sun Nuclear Co., USA) and SNC patient (Sun Nuclear Co., USA Ver 6.1.2.18513). The difference between the coaxial and the universal VMAT plans are follow. The coaxial VMAT plan is better in the V10Gy of OAR, Up to 4.1%, at least 0.4%, the average difference was 1.9% and In the D0.03cc of OAR, Up to 83.6 cGy, at least 2.2 cGy, the average difference was 33.3 cGy. In Dmean, Up to 34.8 cGy, at least -13.0 cGy, the average difference was 9.6 cGy that say the coaxial VMAT plans are better except few cases. H.I difference Up to 0.04, at least 0.01, the average difference was 0.02 and the difference of average

  17. Genotoxic effects of the carbamate insecticide Pirimor-50® in Vicia faba root tip meristems and human lymphocyte culture after direct application and treatment with its metabolic extracts.

    Science.gov (United States)

    Valencia-Quintana, Rafael; Gómez-Arroyo, Sandra; Sánchez-Alarcón, Juana; Milić, Mirta; Olivares, José Luis Gómez; Waliszewski, Stefan M; Cortés-Eslava, Josefina; Villalobos-Pietrini, Rafael; Calderón-Segura, María Elena

    2016-12-01

    The aim of the study was to evaluate genotoxic effects of Pirimor-50®, a pirimicarb-based formulation (50 % active ingredient), in human lymphocyte cultures and Vicia faba root meristems. Furthermore, the objective was to examine a combined influence of insecticide treatment with mammalian microsomal S9 and vegetal S10 metabolic fractions or S10 mix metabolic transformation extracts (after Vicia faba primary roots treatment with Pirimor-50®). We used sister chromatid exchange assay-SCE and measured cell cycle progression and proliferation (proportion of M1-M3 metaphases and replication index ratio-RI). Two processes were used for plant promutagen activation: in vivo activation-Pirimor-50® was applied for 4 h to the plant and then S10 mix was added to lymphocytes; and, in vitro activation-lymphocytes were treated with Pirimor-50® and S10 or S9 for 2 h. Direct treatment induced significantly higher SCE frequencies in meristems at 0.01 mg mL-1. In lymphocytes, significantly higher SCE was at 1 mg mL-1 with decrease in RI and M1-M3 metaphase proportions at 0.5 mg mL-1 and cell division stop at 2.5 mg mL1. S10 mix lymphocyte treatment showed significantly elevated SCE values at 2-2.5 mg mL-1, with cell death at 3 mg mL-1. Lymphocyte treatment with Pirimor-50® together with S9 or S10 showed slightly elevated SCE frequency but had a significant influence on RI decrease, with lowest values in S9 treatment. Since no data are available on the genotoxicity of Pirimor-50®, this study is one of the first to evaluate and compare its direct effect in two bioassays, animal and vegetal, and also the effect of plant and animal metabolism on its genotoxic potential.

  18. Are simple IMRT beams more robust against MLC error? Exploring the impact of MLC errors on planar quality assurance and plan quality for different complexity beams.

    Science.gov (United States)

    Wang, Jiazhou; Jin, Xiance; Peng, Jiayuan; Xie, Jiang; Chen, Junchao; Hu, Weigang

    2016-01-01

    This study investigated the impact of beam complexities on planar quality assur-ance and plan quality robustness by introducing MLC errors in intensity-modulate radiation therapy. Forty patients' planar quality assurance (QA) plans were enrolled in this study, including 20 dynamic MLC (DMLC) IMRT plans and 20 static MLC (SMLC) IMRT plans. The total beam numbers were 150 and 160 for DMLC and SMLC, respectively. Six different magnitudes of MLC errors were introduced to these beams. Gamma pass rates were calculated by comparing error-free fluence and error-induced fluence. The plan quality variation was acquired by comparing PTV coverage. Eight complexity scores were calculated based on the beam flu-ence and the MLC sequence. The complexity scores include fractal dimension, monitor unit, modulation index, fluence map complexity, weighted average of field area, weighted average of field perimeter, and small aperture ratio (plan quality varia-tion. For planar QA, the most significant complexity index was fractal dimension for DMLC (p = -0.40) and weighted segment area for SMLC (p = 0.27) at low magnitude MLC error. For plan quality, the most significant complexity index was weighted segment perimeter for DMLC (p = 0.56) and weighted segment area for SMLC (p= 0.497) at low magnitude MLC error. The sensitivity of planar QA was weakly associated with the field complexity with low magnitude MLC error, but the plan quality robustness was associated with beam complexity. Plans with simple beams were more robust to MLC error.

  19. Dynamic tumor tracking using the Elekta Agility MLC

    Energy Technology Data Exchange (ETDEWEB)

    Fast, Martin F., E-mail: martin.fast@icr.ac.uk; Nill, Simeon, E-mail: simeon.nill@icr.ac.uk; Bedford, James L.; Oelfke, Uwe [Joint Department of Physics, The Institute of Cancer Research and The Royal Marsden NHS Foundation Trust, London SM2 5NG (United Kingdom)

    2014-11-01

    Purpose: To evaluate the performance of the Elekta Agility multileaf collimator (MLC) for dynamic real-time tumor tracking. Methods: The authors have developed a new control software which interfaces to the Agility MLC to dynamically program the movement of individual leaves, the dynamic leaf guides (DLGs), and the Y collimators (“jaws”) based on the actual target trajectory. A motion platform was used to perform dynamic tracking experiments with sinusoidal trajectories. The actual target positions reported by the motion platform at 20, 30, or 40 Hz were used as shift vectors for the MLC in beams-eye-view. The system latency of the MLC (i.e., the average latency comprising target device reporting latencies and MLC adjustment latency) and the geometric tracking accuracy were extracted from a sequence of MV portal images acquired during irradiation for the following treatment scenarios: leaf-only motion, jaw + leaf motion, and DLG + leaf motion. Results: The portal imager measurements indicated a clear dependence of the system latency on the target position reporting frequency. Deducting the effect of the target frequency, the leaf adjustment latency was measured to be 38 ± 3 ms for a maximum target speed v of 13 mm/s. The jaw + leaf adjustment latency was 53 ± 3 at a similar speed. The system latency at a target position frequency of 30 Hz was in the range of 56–61 ms for the leaves (v ≤ 31 mm/s), 71–78 ms for the jaw + leaf motion (v ≤ 25 mm/s), and 58–72 ms for the DLG + leaf motion (v ≤ 59 mm/s). The tracking accuracy showed a similar dependency on the target position frequency and the maximum target speed. For the leaves, the root-mean-squared error (RMSE) was between 0.6–1.5 mm depending on the maximum target speed. For the jaw + leaf (DLG + leaf) motion, the RMSE was between 0.7–1.5 mm (1.9–3.4 mm). Conclusions: The authors have measured the latency and geometric accuracy of the Agility MLC, facilitating its future use for clinical

  20. Complete transformation of ZnO and CuO nanoparticles in culture medium and lymphocyte cells during toxicity testing.

    Science.gov (United States)

    Ivask, Angela; Scheckel, Kirk G; Kapruwan, Pankaj; Stone, Vicki; Yin, Hong; Voelcker, Nicolas H; Lombi, Enzo

    2017-03-01

    Here, we present evidence on complete transformation of ZnO and CuO nanoparticles, which are among the most heavily studied metal oxide particles, during 24 h in vitro toxicological testing with human T-lymphocytes. Synchrotron radiation-based X-ray absorption near edge structure (XANES) spectroscopy results revealed that Zn speciation profiles of 30 nm and 80 nm ZnO nanoparticles, and ZnSO4- exposed cells were almost identical with the prevailing species being Zn-cysteine. This suggests that ZnO nanoparticles are rapidly transformed during a standard in vitro toxicological assay, and are sequestered intracellularly, analogously to soluble Zn. Complete transformation of ZnO in the test conditions was further supported by almost identical Zn spectra in medium to which ZnO nanoparticles or ZnSO4 was added. Likewise, Cu XANES spectra for CuO and CuSO4-exposed cells and cell culture media were similar. These results together with our observation on similar toxicological profiles of ZnO and soluble Zn, and CuO and soluble Cu, underline the importance of dissolution and subsequent transformation of ZnO and CuO nanoparticles during toxicological testing and provide evidence that the nano-specific effect of ZnO and CuO nanoparticles is negligible in this system. We strongly suggest to account for this aspect when interpreting the toxicological results of ZnO and CuO nanoparticles.

  1. In vivo synergistic cytogenetic effects of aminophylline on lymphocyte cultures from patients with lung cancer undergoing chemotherapy

    Energy Technology Data Exchange (ETDEWEB)

    Mylonaki, Effie; Manika, Katerina [Pulmonary Department, “G.Papanikolaou” General Hospital, Aristotle University of Thessaloniki (Greece); Zarogoulidis, Paul, E-mail: pzarog@hotmail.com [Pulmonary Department, “G.Papanikolaou” General Hospital, Aristotle University of Thessaloniki (Greece); Domvri, Kalliopi; Voutsas, Vasilis; Zarogoulidis, Kostas [Pulmonary Department, “G.Papanikolaou” General Hospital, Aristotle University of Thessaloniki (Greece); Mourelatos, Dionysios [Biology and Genetics, Medical School, Aristotle University of Thessaloniki (Greece)

    2012-12-15

    Highlights: ► SCEs in vivo, a possible predictor of tumor chemoresponse. ► In vivo exposure to combined treatment, applying the SCE assay. ► Aminophylline enhances DNA instability induced by chemotherapy in vivo. ► In vivo synergistic effect of Aminophylline with the chemotherapeutic agents. - Abstract: Background: The anti-cancer and cytogenetic effects of aminophylline (AM) have been demonstrated in several clinical trials. The aim of the present study was to investigate the in vivo cytogenetic effects of AM in newly diagnosed patients with small cell (SCLC) and non-small cell lung cancer (NSCLC), receiving chemotherapy for the first time. Methods: Sister chromatid exchanges (SCEs) and proliferation rate index (PRI) were evaluated in peripheral blood lymphocyte cultures from six patients with SCLC and six patients with NSCLC after the in vitro addition of AM and after the in vivo administration of AM in patients receiving chemotherapy. Results: The in vitro addition of AM significantly increased SCEs only in SCLC patients (p < 0.001). The in vivo administration of AM after chemotherapy increased SCEs in both cancer types (SCLC: p < 0.001, NSCLC: p = 0.003) and this increase was synergistic, the rates of SCEs in the presence of AM were higher than the expected SCE values if the increases above background for chemotherapy and AM were independent and additive (SCLC: p < 0.001, NSCLC: p = 0.008). Although in both groups of patients cell division delays were observed after the combined chemotherapy plus in vivo AM treatment, the correlation between the magnitude of the SCE response and the PRI depression was not statistically significant (p > 0.05). Conclusions: These observations suggest that AM enhances the results of concurrently administered chemotherapy by synergistically increasing its cytogenetic effects in patients with lung cancer.

  2. Proliferative kinetics and chromosome damage in trisomy 21 lymphocyte cultures exposed to gamma-rays and bleomycin

    Energy Technology Data Exchange (ETDEWEB)

    Morimoto, K.; Kaneko, T.; Iijima, K.; Koizumi, A.

    1984-04-01

    Lymphocytes from patients with Down's syndrome (trisomy 21) have been investigated for cell cycle kinetics, cell proliferation delays, and chromosomal aberrations after exposure to gamma-rays or bleomycin. Analysis by sister chromatid differential staining revealed that trisomy 21 lymphocytes started cell cycling about 5 hr earlier than did normal diploid lymphocytes after phytohemagglutinin stimulation as a whole, but that cycling trisomic and normal cells had the same mean cell cycle times. When exposed to gamma-rays or bleomycin in G0, trisomy 21 lymphocytes showed a 30% or, on average, 50% longer duration of cell turnover times, respectively, than normal cells; only bleomycin-treated trisomic cells had a biphasic dose-response. Frequencies of dicentrics and rings in first-division cells after gamma-ray or bleomycin exposure were twice as high in trisomic cells as in normal cells. The frequency of aberrations decreased by 50% (gamma-ray-exposed) or 65 to 85% (bleomycin-treated) through successive divisions; trisomic cells showed a more marked decline in aberration yields compared to normal cells after bleomycin treatment. These data support the idea that circulating lymphocytes in trisomy 21 patients have a shorter average life span or a younger average age.

  3. Implementation and acceptance of dynamic MLC for IMRT and VMAT; Implementacion y aceptacion de MLC dinamicos para IMRT y VMAT

    Energy Technology Data Exchange (ETDEWEB)

    Garcia, B.; Marquina, J.; Ramirez, J.; Gonzales, A., E-mail: bertha.garcia@aliada.com.pe [ALIADA, Oncologia Integral, Av. Jose Galvez Barrenechea 1044, San Isidro, Lima 27 (Peru)

    2014-08-15

    The use of Multi-leaf Collimator (MLC) in Intensity-Modulated Radiation Therapy (IMRT) for dynamic treatment techniques as Volumetric Modulated Arc Therapy (VMAT) makes that the movement controls as the speed of the MLC are quantified by means of an acceptance test. The objective determination of the operation regulations of the radiotherapy equipment requires ideally the establishment of the quantitative relationship among the performance deviations and clinical results or some acceptable substitute. Different protocols exist detailed with parameters and acceptance ranges according to the MLC thickness. In our case the acceptance test was carried out for 120-MLC of Trilogy equipment brand Varian. For all the test were used 300-200 Um for each formed beam lets; source-surface distance (SSD) of 100 cm. 9 acceptance tests were used each one with different purposes like to quantify, synchronization, stability, complexity, precision, positioning, conformity, dynamic movements for the case of dynamic wedges, consecutive moves, among others, for the measurements were used film badges dosimetry (Gafchromic Ebt-3 scanner Epson expression 10000 XL); additionally the results were compared with a diodes arrangement Map-Check 2 brand Sun Nuclear; that consists of 1527 diodes prepared in a field of 32 x 26 cm located at a distance of 1 cm parallel, 0.5 cm diagonally. All the developed tests were inside the acceptance tolerance parameters when comparing the obtained result regarding the badges and the Map-Check was found a discrepancy of 0.01%, what gives a treatment certainty to the moment to impart volumetric dose in dynamic fields to the patients. (Author)

  4. An association, in adult Japanese, between the occurrence of rogue cells among cultured lymphocytes (JC virus activity) and the frequency of simple chromosomal damage among the lymphocytes of persons exhibiting these rogue cells

    Energy Technology Data Exchange (ETDEWEB)

    Neel, J.V. [Univ. of Michigan, Ann Arbor, MI (United States). Dept. of Human Genetics

    1998-08-01

    Data from a previous study of the cytogenetic effects, in cultured lymphocytes, of exposure to the atomic bomb in Hiroshima have been reanalyzed to determine the relationship between the occurrence of rogue cells in an individual and the frequency of simple chromosomal damage in the nonrogue cells of the same individual. Rogue cells are cells with complex chromosomal damage, currently believed to be a manifestation of the activity of a human polyoma virus termed JC. Among a total of 1,835 persons examined, there were 45 exhibiting rogue cells. A total of 179,599 cells were scored for simple chromosomal damage. In both the exposed and the control populations, there was an absolute increase of {approximately}1.5% in the frequency of simple chromosomal damage in the nonrogue cells of those exhibiting rogue cells, when compared with the frequencies observed in those not exhibiting rogue cells, which is a statistically significant difference. It is argued that this phenomenon, occurring not only in lymphocytes but possibly also in other cells/tissues, may play a contributory role in the origin of malignancies characterized by clonal chromosome abnormalities. Unexpectedly, among those exhibiting rogue cells, there was a disproportionately greater representation of persons who had received relatively high radiation exposures from the bomb. The reason for this is unclear, but it is tempting to relate the finding to some lingering effect of the exposure (or the circumstances surrounding the exposure) on immunocompetence.

  5. A Novel MLC Scheme with M2-QAM Constellations over AWGN and Rayleigh Fading Channels

    Institute of Scientific and Technical Information of China (English)

    YUANDongfeng; ZHANGPeng; WayneE.Stark

    2003-01-01

    Optimal multilevel codes (MLC) with M2-QAM constellations are considered with focus on both AWGN and Rayleigh fading channels. With the proposition of three new set partitioning rules for M2-QAM constellations, a novel simplified MLC scheme with a very simple MSD (Multistage decoding) structure comes into being, which is operated on the capacity rule. Compared with traditional MLC/MSD scheme the new MLC/MSD structure greatly reduces the calculation of the individual capacities in MLC system because the new set partitioning rules ensure the independency of two dimensions symbols. Furthermore the new MLC/MSD system cannot enhance any performance loss and the time delay is just 1/2 of that with traditional partitioning rules. As an example 64-QAM constellation with three new set partitioning rules are provided. Their performance is simulated over AWGN and Rayleigh fading channels by choosing BCH codes as the component codes.

  6. An experimental evaluation of the Agility MLC for motion-compensated VMAT delivery

    Science.gov (United States)

    Davies, G. A.; Clowes, P.; Bedford, J. L.; Evans, P. M.; Webb, S.; Poludniowski, G.

    2013-07-01

    An algorithm for dynamic multileaf-collimator (dMLC) tracking of a target performing a known a priori, rigid-body motion during volumetric modulated arc therapy (VMAT), has been experimentally validated and applied to investigate the potential of the Agility (Elekta AB, Stockholm, Sweden) multileaf-collimator (MLC) for use in motion-compensated VMAT delivery. For five VMAT patients, dosimetric measurements were performed using the Delta4 radiation detector (ScandiDos, Uppsala, Sweden) and the accuracy of dMLC tracking was evaluated using a gamma-analysis, with threshold levels of 3% for dose and 3 mm for distance-to-agreement. For a motion trajectory with components in two orthogonal directions, the mean gamma-analysis pass rate without tracking was found to be 58.0%, 59.0% and 60.9% and was increased to 89.1%, 88.3% and 93.1% with MLC tracking, for time periods of motion of 4 s, 6 s and 10 s respectively. Simulations were performed to compare the efficiency of the Agility MLC with the MLCi MLC when used for motion-compensated VMAT delivery for the same treatment plans and motion trajectories. Delivery time increases from a static-tumour to dMLC-tracking VMAT delivery were observed in the range 0%-20% for the Agility, and 0%-57% with the MLCi, indicating that the increased leaf speed of the Agility MLC is beneficial for MLC tracking during lung radiotherapy.

  7. Complete transformation of ZnO and CuO nanoparticles in culture medium and lymphocyte cells during toxicity testing

    Science.gov (United States)

    Here, we present evidence on complete transformation of ZnO and CuO nanoparticles, which are among the most heavily studied metal oxide particles, during 24 h in vitro toxicological testing with human T-lymphocytes. Synchrotron radiation-based X-ray absorption near edge st...

  8. Effects of age on segregation of the X and Y chromosomes in cultured lymphocytes from Chinese men.

    Science.gov (United States)

    Song, Yaxian; Chen, Qian; Zhang, Zhen; Hou, Heli; Zhang, Ding; Shi, Qinghua

    2009-08-01

    Chromosome malsegregation in binucleated lymphocytes is a useful endpoint to evaluate age effect on genetic stability. However, the investigations on chromosome malsegregation in binucleated lymphocytes from Chinese are scarce. In this study, peripheral blood lymphocytes were collected from 14 old (60-70 years) and 10 young (22-26 years) healthy Chinese men. To detect malsegregation of the sex chromosomes, multi-color fluorescence in situ hybridization (FISH) was performed on binucleated lymphocytes, cytokinesis-blocked by cytochalasin B at the first mitosis after phytohaemagglutinin stimulation. Compared with that in young men, a significant increase in frequencies of loss of chromosome X (9.2 +/- 3.2 per thousand vs. 1.1 +/- 0.9 per thousand, P men. Similarly, nondisjunction of chromosome X (16.5 +/- 3.4 per thousand vs. 3.5 +/- 1.1 per thousand, P men than in young men. Regardless of donor's age, nondisjunction is more prevalent than loss for both chromosome X and Y. The frequencies of observed simultaneous malsegregation were relatively higher than the expected, suggesting an association between malsegregation. These results indicated that in Chinese men, malsegregation of the sex chromosomes increases with age in an associated fashion, and nondisjunction accounts for the majority of spontaneous chromosome malsegregation.

  9. Pennogenin tetraglycoside induces rat myometrial contraction and MLC20 phosphorylation via PLC-IP(3 and RhoA/Rho kinase signaling pathways.

    Directory of Open Access Journals (Sweden)

    Limei Wang

    Full Text Available BACKGROUND: Total steroidal saponins extracted from the rhizome of Paris polyphylla Sm. var. yunnanensis (TSSPs have been widely used in China for the treatment of abnormal uterine bleeding. We previously studied the main active constituents of TSSPs and their structure-activity relationships with respect to rat myometrial contractions. Tg (pennogenin tetraglycoside was identified as one of the active ingredients in TSSPs able to induce rat myometrial contractions. However, the mechanisms underlying the pharmacological actions on uterine activity have not been described clearly. METHODS: Here Tg was screened for effects on contractile activity in isolated uterine strips from estrogen-primed rats and on MLC20 phosphorylation and related signaling pathways in cultured rat myometrial cells as determined by Western blot. Intracellular calcium ([Ca(2+](i was monitored under a confocal microscope using Fluo-4 AM-loaded myometrial cells. RESULTS: Tg dose-dependently stimulated rat myometrial contractions as well as MLC20 phosphorylation in vitro, which could be completely suppressed by an inhibitor of myosin light chain kinase (MLCK. Use of Ca(2+ channel blockers and kinase inhibitors demonstrated that Tg-induced myometrial contractions are mediated by activation of the phospholipase C (PLC-inositol triphosphate (IP3 signaling pathway, resulting in increased MLC20 phosphorylation. Furthermore, Y27632, a specific inhibitor of Rho kinase (ROK, notably suppressed Tg-stimulated myometrial contractions and decreased MLC20 phosphorylation. CONCLUSIONS: These data provide evidence that rat myometrial contractility induced by Tg results from enhanced MLC20 phosphorylation, while both PLC-IP3 and RhoA/ROK signaling pathways mediate the process. These mechanisms may be responsible for the therapeutic effects of TSSPs on abnormal uterine bleeding.

  10. MHC class I phenotype and function of human beta 2-microglobulin transgenic murine lymphocytes

    DEFF Research Database (Denmark)

    Bjerager, L; Pedersen, L O; Bregenholt, S;

    1996-01-01

    Lymphoid cells from beta 2-microglobulin (beta 2m) knockout mice transgenic for human (h) beta 2m (C57BL/10 m beta 2m-/h beta 2m+) were compared with normal mice for their binding to exogenously added h beta 2m, binding to a H-2Db peptide and for functional activity in a one-way allogenic MLC....... Based on data from cellular binding studies, Scatchard analyses and flow cytometry, it is concluded that exogenous h beta 2m does not bind to hybrid MHC class I (MHC-I) molecules composed of mouse heavy chain/h beta 2m molecules expressed on lymphocytes of transgenic mice. Immunoprecipitation and SDS...... binds radiolabelled peptide in the absence of exogenous added h beta 2m suggesting that a stable fraction of hybrid H-2Db molecules is empty or contain peptides with very low affinity. In a one-way allogenic mixed lymphocyte culture, transgenic splenocytes were found to be far less stimulatory than...

  11. SIEMENS MLC-82日常维护及故障检修%Routine maintenance and trouble shooting of SIEMENS MLC-82 multi-leaf collimator

    Institute of Scientific and Technical Information of China (English)

    陶长龙

    2011-01-01

    The MLC-82 electric multi-leaf collimator (MLC-Multileaf Collimator) system of Siemens is composed of two parts of hardware and software. In this paper, the author introduced the effect of the collimator and methods for maintenance of its hardware to share with counterparts.%西门子MLC-82电动多叶准直器(MLC—MultileafCollimator)系统由硬件和软件两部分组成。简要阐述了多叶准直器的作用,系统硬件的维修经验,介绍SIEMENS MLC-82硬件系统的日常维护及常见故障的检修。

  12. Impact of MLC properties and IMRT technique in meningioma and head-and-neck treatments

    DEFF Research Database (Denmark)

    Kantz, Steffi; Söhn, Matthias; Troeller, Almut

    2015-01-01

    -planned for step-and-shoot IMRT (ssIMRT), sliding window IMRT (dMLC) and VMAT using the MLCi2 without (-) and with (+) interdigitation and the Agility-MLC attached to an Elekta 6MV linac. This results in nine plans per patient. Consistent patient individual optimization parameters are used. Plans are generated...

  13. Combining MLC and SVM Classifiers for Learning Based Decision Making: Analysis and Evaluations.

    Science.gov (United States)

    Zhang, Yi; Ren, Jinchang; Jiang, Jianmin

    2015-01-01

    Maximum likelihood classifier (MLC) and support vector machines (SVM) are two commonly used approaches in machine learning. MLC is based on Bayesian theory in estimating parameters of a probabilistic model, whilst SVM is an optimization based nonparametric method in this context. Recently, it is found that SVM in some cases is equivalent to MLC in probabilistically modeling the learning process. In this paper, MLC and SVM are combined in learning and classification, which helps to yield probabilistic output for SVM and facilitate soft decision making. In total four groups of data are used for evaluations, covering sonar, vehicle, breast cancer, and DNA sequences. The data samples are characterized in terms of Gaussian/non-Gaussian distributed and balanced/unbalanced samples which are then further used for performance assessment in comparing the SVM and the combined SVM-MLC classifier. Interesting results are reported to indicate how the combined classifier may work under various conditions.

  14. Combining MLC and SVM Classifiers for Learning Based Decision Making: Analysis and Evaluations

    Directory of Open Access Journals (Sweden)

    Yi Zhang

    2015-01-01

    Full Text Available Maximum likelihood classifier (MLC and support vector machines (SVM are two commonly used approaches in machine learning. MLC is based on Bayesian theory in estimating parameters of a probabilistic model, whilst SVM is an optimization based nonparametric method in this context. Recently, it is found that SVM in some cases is equivalent to MLC in probabilistically modeling the learning process. In this paper, MLC and SVM are combined in learning and classification, which helps to yield probabilistic output for SVM and facilitate soft decision making. In total four groups of data are used for evaluations, covering sonar, vehicle, breast cancer, and DNA sequences. The data samples are characterized in terms of Gaussian/non-Gaussian distributed and balanced/unbalanced samples which are then further used for performance assessment in comparing the SVM and the combined SVM-MLC classifier. Interesting results are reported to indicate how the combined classifier may work under various conditions.

  15. Optimisation of the CT h4S bioassay for detection of human interleukin-4 secreted by mononuclear cells stimulated by phytohaemaglutinin or by human leukocyte antigen mismatched mixed lymphocyte culture

    DEFF Research Database (Denmark)

    Petersen, Søren Lykke; Russell, Charlotte Astrid; Bendtzen, Klaus;

    2002-01-01

    bioassay with regards to specificity, sensitivity, detection limit, and reproducibility. We have found the optimal assay conditions to be 1 x 10 (4) CT.h4S cells/well deprived of IL-4 for 24 h and preincubated for 7 h followed by 18 h of incubation with tritiated methyl-thymidine. In this setting the CT.h4...... of IL-4 detection was not due to high amounts of soluble IL-4 receptor. With the use of 1x10(6) responder cells/well in HLA-mismatched MLC, we found limited IL-4 accumulation still increasing at day 12. We conclude that the CT.h4S bioassay is a reliable and specific method for quantification of IL-4......Limiting dilution analysis has been used in the context of allogeneic bone marrow transplantation to determine anti-recipient interleukin-2 (IL-2) producing helper T lymphocyte precursor (HTLp) frequencies, which in several studies have been predictive of graft-versus-host disease (GVHD). Recently...

  16. Chromosome aberrations and sister chromatid exchanges in cultured human lymphocytes treated with sodium metabisulfite, a food preservative.

    Science.gov (United States)

    Rencüzogullari, E; Ila, H B; Kayraldiz, A; Topaktaş, M

    2001-02-20

    The aim of this study was to investigate the ability of sodium metabisulfite (SMB) which is used as an antimicrobial substance in food, to induce chromosome aberrations (CA) and sister chromatid exchanges (SCE) in human lymphocytes. SMB-induced CAs and SCEs at all concentrations (75, 150 and 300 microg/ml) and treatment periods (24 and 48h) dose-dependently. However, SMB decreased the replication index (RI) and the mitotic index (MI) at the concentrations of 150 and 300 microg/ml for 24 and 48h treatment periods. This decrease was dose-dependent as well.

  17. Evaluation of dosimetric effect caused by slowing with multi-leaf collimator (MLC leaves for volumetric modulated arc therapy (VMAT

    Directory of Open Access Journals (Sweden)

    Xu Zhengzheng

    2016-03-01

    Full Text Available This study is to report 1 the sensitivity of intensity modulated radiation therapy (IMRT QA method for clinical volumetric modulated arc therapy (VMAT plans with multi-leaf collimator (MLC leaf errors that will not trigger MLC interlock during beam delivery; 2 the effect of non-beam-hold MLC leaf errors on the quality of VMAT plan dose delivery.

  18. Inhibitory effects of telmisartan on culture and proliferation of and Kv1.3 potassium channel expression in peripheral blood CD4+ T lymphocytes from Xinjiang Kazakh patients with hypertension

    Directory of Open Access Journals (Sweden)

    Sha-Sha Huang

    2016-10-01

    Full Text Available Introduction: Activation of T lymphocytes, for which potassium channels are essential, is involved in the development of hypertension. In this study, we explored the inhibitory effects of telmisartan on the culture and proliferation of and Kv1.3 potassium channel expression in peripheral blood CD4+ T lymphocytes derived from Xinjiang Kazakh patients with hypertension. Methods: CD4+ T-cell samples from hypertensive Kazakh patients and healthy Kazakh people were divided into healthy control, case control, telmisartan, and 4-aminopytidine groups. Changes in the expression levels of interleukin (IL-6 and IL-17 in the blood of the healthy control and case control subjects were detected by enzyme-linked immunosorbent assay. Peripheral blood CD4+ T lymphocytes were first activated and proliferated in vitro and then incubated for 0, 24, and 48 h under various treatment conditions. Thereafter, changes in CD4+ T-lymphocytic proliferation were determined using Cell Counting Kit-8 and microscope photography. Changes in messenger RNA (mRNA and protein expression of the Kv1.3 potassium channel in CD4+ T lymphocytes were detected using real-time quantitative polymerase chain reaction and Western blots, respectively. Results: The IL-6 and IL-17 expression levels were significantly higher in the blood of the hypertensive Kazakh patients than in the healthy Kazakh people. Telmisartan inhibited T-lymphocytic proliferation, as well as the mRNA and protein expression of the Kv1.3 potassium channel in CD4+ T lymphocytes, and the inhibitory effects were time-dependent, with the strongest inhibition observed after 48 h and significantly weaker inhibition observed after 24 h of treatment. Conclusions: Telmisartan may potentially regulate hypertensive inflammatory responses by inhibiting T-lymphocytic proliferation and Kv1.3 potassium channel expression in CD4+ T lymphocytes.

  19. Inhibitory effects of telmisartan on culture and proliferation of and Kv1.3 potassium channel expression in peripheral blood CD4+ T lymphocytes from Xinjiang Kazakh patients with hypertension.

    Science.gov (United States)

    Huang, Sha-Sha; Zhang, Qiu-Bing; Yuan, Qing-Yan; He, Si-Li; Zhang, Yuan-Ming

    2016-10-01

    Activation of T lymphocytes, for which potassium channels are essential, is involved in the development of hypertension. In this study, we explored the inhibitory effects of telmisartan on the culture and proliferation of and Kv1.3 potassium channel expression in peripheral blood CD4(+) T lymphocytes derived from Xinjiang Kazakh patients with hypertension. CD4(+) T-cell samples from hypertensive Kazakh patients and healthy Kazakh people were divided into healthy control, case control, telmisartan, and 4-aminopytidine groups. Changes in the expression levels of interleukin (IL)-6 and IL-17 in the blood of the healthy control and case control subjects were detected by enzyme-linked immunosorbent assay. Peripheral blood CD4(+) T lymphocytes were first activated and proliferated in vitro and then incubated for 0, 24, and 48 h under various treatment conditions. Thereafter, changes in CD4(+) T-lymphocytic proliferation were determined using Cell Counting Kit-8 and microscope photography. Changes in messenger RNA (mRNA) and protein expression of the Kv1.3 potassium channel in CD4(+) T lymphocytes were detected using real-time quantitative polymerase chain reaction and Western blots, respectively. The IL-6 and IL-17 expression levels were significantly higher in the blood of the hypertensive Kazakh patients than in the healthy Kazakh people. Telmisartan inhibited T-lymphocytic proliferation, as well as the mRNA and protein expression of the Kv1.3 potassium channel in CD4(+) T lymphocytes, and the inhibitory effects were time-dependent, with the strongest inhibition observed after 48 h and significantly weaker inhibition observed after 24 h of treatment. Telmisartan may potentially regulate hypertensive inflammatory responses by inhibiting T-lymphocytic proliferation and Kv1.3 potassium channel expression in CD4(+) T lymphocytes. © The Author(s) 2016.

  20. Comparison of differences between dicentric assay and translocation analysis for biodosimetry in cultured peripheral blood lymphocytes of Korean individuals

    Energy Technology Data Exchange (ETDEWEB)

    Park, Hyun Jin; Park, Mi Young; Seo, Min Ji; Kwon, Hee Kyung; Lee, Su Jae; Lee, Yun Sil; Ji, Young Hoon; Choi, Soo Yong; Cho, Chul Koo; Kim, Tae Hwan; Kang, Chang Mo [Korea Institute of Radiological and Medical Sciences, Seoul (Korea, Republic of)

    2004-07-01

    Chromosome aberrations are considered to be important indicators of induced DNA damage and genomic instability. For this reason, they constitute the main parameter used to monitor individuals exposed to radiation. Biological dosimetry using the analysis of dicentrics in human lymphocytes is well established, especially in case of acute exposure, when the blood samples are taken within a few weeks. However, dicentric analysis is not an adequate parameter in case of chronic exposure, because these aberrations are unstable with time, and have a limited use for dose assessment of past exposures. In contrast to dicentrics, however, translocations are considered stable in cell division and so the yield should not fall with time. In the present study, using FISH-chromosome painting analysis with the dose-response curve for chromosome aberrations, we monitored the stable and unstable chromosome aberrations of 2 Korean's periperal blood lymphocytes irradiated in vitro with {gamma}-rays from {sup 137}Cs (doses between 0.0 and 2.0 Gy). By using the dose-response curve for chromosome aberration, our aim was to estimate the absorbed doses, and then establish comparison with the results obtained by conventional dicentric analysis, thus taking the opportunity to test the validity of chromosome aberration analysis by FISH painting method for retrospective biodosimetry in Korean individual.

  1. Establishment of a heterotypic 3D culture system to evaluate the interaction of TREG lymphocytes and NK cells with breast cancer.

    Science.gov (United States)

    Augustine, Tanya N; Dix-Peek, Thérèse; Duarte, Raquel; Candy, Geoffrey P

    2015-11-01

    Three-dimensional (3D) culture approaches to investigate breast tumour progression are yielding information more reminiscent of the in vivo microenvironment. We have established a 3D Matrigel system to determine the interactions of luminal phenotype MCF-7 cells and basal phenotype MDA-MB-231 cells with regulatory T lymphocytes and Natural Killer cells. Immune cells were isolated from peripheral blood using magnetic cell sorting and their phenotype validated using flow cytometry both before and after activation with IL-2 and phytohaemagglutinin. Following the establishment of the heterotypic culture system, tumour cells displayed morphologies and cell-cell associations distinct to that observed in 2D monolayer cultures, and associated with tissue remodelling and invasion processes. We found that the level of CCL4 secretion was influenced by breast cancer phenotype and immune stimulation. We further established that for RNA extraction, the use of proteinase K in conjunction with the Qiagen RNeasy Mini Kit and only off-column DNA digestion gave the best RNA yield, purity and integrity. We also investigated the efficacy of the culture system for immunolocalisation of the biomarkers oestrogen receptor-α and the glycoprotein mucin 1 in luminal phenotype breast cancer cells; and epidermal growth factor receptor in basal phenotype breast cancer cells, in formalin-fixed, paraffin-wax embedded cultures. The expression of these markers was shown to vary under immune mediation. We thus demonstrate the feasibility of using this co-culture system for downstream applications including cytokine analysis, immunolocalisation of tumour biomarkers on serial sections and RNA extraction in accordance with MIQE guidelines.

  2. Cultured lymphocytes from autistic children and non-autistic siblings up-regulate heat shock protein RNA in response to thimerosal challenge.

    Science.gov (United States)

    Walker, Stephen J; Segal, Jeffrey; Aschner, Michael

    2006-09-01

    There are reports suggesting that some autistic children are unable to mount an adequate response following exposure to environmental toxins. This potential deficit, coupled with the similarity in clinical presentations of autism and some heavy metal toxicities, has led to the suggestion that heavy metal poisoning might play a role in the etiology of autism in uniquely susceptible individuals. Thimerosal, an anti-microbial preservative previously added routinely to childhood multi-dose vaccines, is composed of 49.6% ethyl mercury. Based on the levels of this toxin that children receive through routine immunization schedules in the first years of life, it has been postulated that thimerosal may be a potential triggering mechanism contributing to autism in susceptible individuals. One potential risk factor in these individuals may be an inability to adequately up-regulate metallothionein (MT) biosynthesis in response to presentation of a heavy metal challenge. To investigate this hypothesis, cultured lymphocytes (obtained from the Autism Genetic Resource Exchange, AGRE) from autistic children and non-autistic siblings were challenged with either 10 microM ethyl mercury, 150 microM zinc, or fresh media (control). Following the challenge, total RNA was extracted and used to query "whole genome" DNA microarrays. Cultured lymphocytes challenged with zinc responded with an impressive up-regulation of MT transcripts (at least nine different MTs were over-expressed) while cells challenged with thimerosal responded by up-regulating numerous heat shock protein transcripts, but not MTs. Although there were no apparent differences between autistic and non-autistic sibling responses in this very small sampling group, the differences in expression profiles between those cells treated with zinc versus thimerosal were dramatic. Determining cellular response, at the level of gene expression, has important implications for the understanding and treatment of conditions that result

  3. DVH Analysis of Cobalt-60 treatment plans incorporating a recently developed MLC

    Directory of Open Access Journals (Sweden)

    Gangarapu Sri Krishna

    2016-07-01

    Full Text Available Purpose: The aim of this investigation was to measure the gain in DVH indices when the recently developed MLC was used for Cobalt-60 treatments.Methods: A prototype multileaf collimator (MLC that was retrofitted to telecobalt-60 therapy machine was reported and is currently proposed for clinical trials in our institution. Ten patients’ plans that were previously planned through an ECLIPSE® treatment planning system and were treated with open beams from Cobalt-60 machine were imported into Radiation Oncology Planning System [ROPS] and the dose calculations and dose volume histogram (DVH analysis were performed. The plans were re-planned using the Cobalt-MLC, a feature available in the ROPS planning system. The DVH analysis consisted of conformity index (CI, homogeneity index (HI and conformation number (CN. The results of this study are presented in this paper. The analysis specifically aimed at measuring the gain in these indices when the MLC was compared with open beams.Results: DVH Comparison of ten sites using open and Cobalt MLC fields showed that the use of MLC results in reduced normal tissue dose, while maintaining the GTV dose. Lower value of CI for normal structures was observed demonstrating the sparing of critical organs when MLC was used. The index HI was studied to show the significance of hot spots outside the PTV. Hot spots were observed even with MLC beams for some cases due to less number of fields.Conclusion: It has been demonstrated through DVH analysis that the use of the recently developed MLC for Cobalt Teletherapy machine results in benefit for the treatment of patients.

  4. VARIAN 6EX MLC 叶片的维护及故障维修一例

    Institute of Scientific and Technical Information of China (English)

    王良

    2015-01-01

    目的:探讨 VARIAN 6EX MLC 系统叶片基本保养流程的方式和方法。方法:阐述 MLC 系统叶片维护保养的方法以及注意事项,简单介绍 diagLeafBacklashAll 和 diagLeafPWMTestMlc 两个 MLC 叶片测试代码命令的应用。结果:通过掌握 MLC 系统叶片的保养基本流程,可以避免日常使用中频繁的 MLC 故障报错。结论:保证 MLC 系统叶片的及时保养,能够大大减少维修资金和人力成本。

  5. The serological recognition of the human MLC determinants using a modified cytotoxicity technique.

    Science.gov (United States)

    van Rood, J J; van Leeuwen, A; Keuning, J J; van Oud Alblas, A B

    1975-04-01

    A new approach which presumably recognizes MLC determinants (or at least structures closely linked to MLC and different from the FOUR, AJ and LA series determinants) by serology is described. As the test can be performed within 10 h it could be used to match cadaveric kidney donors for these MLC determinants. Perhaps even more important is the fact that the antibodies detected by this technique could be used to study the immunochemistry of a class of determinants differing from the well known SD determinants.

  6. Tumor infiltrating lymphocytes in melanoma comprise high numbers of T-cell clonotypes that are lost during in vitro culture

    DEFF Research Database (Denmark)

    thor Straten, P; Kirkin, A F; Siim, E;

    2000-01-01

    -associated peptide epitopes. Cultured TIL have been studied in order to unveil characteristics of TIL and the interactions of TIL and melanoma cells. Whether in vitro cultured TIL mirrors the in situ situation has, however, been questioned. In the present study we have taken advantage of T-cell receptor clonotype...... mapping methodology to conduct a full and detailed analysis of the T-cell clonotypes in melanoma lesions and in corresponding lines of TIL established in vitro. All melanoma lesions and the corresponding TIL cultures comprised high numbers of T-cell clonotypes, typically in the range of 40 to more than 60...... that in situ T-cell clonotypes in melanoma are not readily expanded in vitro and that the majority of T-cell clonotypes present in cultured TIL are not present in situ....

  7. Role of the MAPK pathway in the observed bystander effect in lymphocytes co-cultured with macrophages irradiated with γ-rays or carbon ions.

    Science.gov (United States)

    Dong, Chen; He, Mingyuan; Ren, Ruiping; Xie, Yuexia; Yuan, Dexiao; Dang, Bingrong; Li, Wenjian; Shao, Chunlin

    2015-04-15

    The radiation-induced bystander effect (RIBE) has potential implications in cancer risks from space particle radiation; however, the mechanisms underlying RIBE are unclear. The role of the MAPK pathway in the RIBEs of different linear energy transfer (LET) was investigated. Human macrophage U937 cells were irradiated with γ-rays or carbon ions and then co-cultured with nonirradiated HMy2.CIR (HMy) lymphocytes for different periods. The activation of MAPK proteins and the generation of intracellular nitric oxide (NO) and reactive oxygen species (ROS) in the irradiated U937 cells were measured. Micronuclei (MN) formation in the HMy cells was applied to evaluate the bystander damage. Some U937 cells were pretreated with different MAPK inhibitors before irradiation. Additional MN formation was induced in the HMy cells after co-culturing with irradiated U937 cells, and the yield of this bystander MN formation was dependent on the co-culture period with γ-ray irradiation but remained high after 1h of co-culture with carbon irradiation. Further investigations disclosed that the time response of the RIBEs had a relationship with LET, where ERK played a different role from JNK and p38 in regulating RIBEs by regulating the generation of the bystander signaling factors NO and ROS. The finding that the RIBE of high-LET radiation could persist for a much longer period than that of γ-rays implies that particle radiation during space flight could have a high risk of long-term harmful effects. An appropriate intervention targeting the MAPK pathway may have significant implications in reducing this risk. Copyright © 2015 Elsevier Inc. All rights reserved.

  8. Avocado fruit (Persea americana Mill) exhibits chemo-protective potentiality against cyclophosphamide induced genotoxicity in human lymphocyte culture.

    Science.gov (United States)

    Paul, Rajkumar; Kulkarni, Paresh; Ganesh, Narayan

    2011-01-01

    Diets rich in fruits and vegetables have been associated with reduced risks for many types of cancers. Avocado (Persea americana Mill.) is a widely consumed fruit containing many cancer preventing nutrients, vitamins and phytochemicals. Studies have shown that phytochemicals extracted from the avocado fruit selectively induce cell cycle arrest, inhibit growth, and induce apoptosis in precancerous and cancer cell lines. Our recent studies indicate that phytochemicals extracted with 50% Methanol from avocado fruits help in proliferation of human lymphocyte cells and decrease chromosomal aberrations induced by cyclophosphamide. Among three concentrations (100 mg, 150 mg and 200 mg per Kg Body Weight), the most effective conc. of extract was 200 mg/Kg Body Wt. It decreased significant level of numerical and structural aberrations (breaks, premature centromeric division etc. up to 88%, p < 0.0001)), and accrocentric associtation within D & G group (up to 78%, p = 0.0008). These studies suggest that phytochemicals from the avocado fruit can be utilized for making active chemoprotective ingredient for lowering the side effect of chemotherapy like cyclophosphamide in cancer therapy.

  9. Assessment of the genotoxicity of imidacloprid and metalaxyl in cultured human lymphocytes and rat bone-marrow.

    Science.gov (United States)

    Demsia, Georgia; Vlastos, Dimitris; Goumenou, Marina; Matthopoulos, Demetrios P

    2007-12-01

    Imidacloprid and metalaxyl are two pesticides that are widely used in agriculture, either separately, or in combination. These agents were studied for their possible genotoxic effects with respect to the following cytogenetic end-points: (1) in vitro micronucleus (MN) formation and sister-chromatid exchange (SCE) induction in human lymphocytes and (2) in vivo micronucleus induction in polychromatic erythrocytes (PCEs) of the rat bone-marrow. The results of the MN analysis indicate that MN frequencies after treatment with both pesticides, separately or as a mixture, do not significantly differ from those in the controls except after treatment with metalaxyl alone at 50 microg/ml (pmetalaxyl at 5, 10 and 100 microg/ml and with the combination of imidacloprid and metalaxyl at 100 and 200 microg/ml. Finally, the in vivo micronucleus assay with rat bone-marrow polychromatic erythrocytes showed a statistically significant effect upon separate treatments with imidacloprid and metalaxyl at doses of 300 mg/kg body weight (b.w.) (p<0.01) or upon combined treatment with 200 mg/Kg b.w. (p<0.001) and 400 mg/kg b.w. (p<0.05).

  10. Implementation of a Quality Control using portal imaging dynamic MLC; Implementacion de un programa de control de calidad de MLC dinamico mediante imagen portal

    Energy Technology Data Exchange (ETDEWEB)

    Sanz Freire, C. J.; Vazquez Galinanes, A.; Collado Chamorro, P. M.; Diaz Pascual, V.; Gomez Amez, J.; Sanchez Martinez, S.; Ossola Lentati, G. A

    2011-07-01

    The precision in the correct beam irradiation in the treatment of highly modulated Intensity Modulated Radiation Therapy (IMRT) depends largely on the characteristics and behavior of the multi leaf collimator (MLC). Quality control (QC) of this element is essential to ensure proper delivery of the beams calculated. It is important to know the absolute position of each sheet, the motion characteristics of each behavior and stability. Among the numerous methods for carrying out the QC MLC, the use of portal imaging is a practical and high resolution. This paper describes the development of a quality control program based dynamic MLC portal image, self-developed software that enables analysis and the results of two years experience following the implementation of IMRT treatments at our center. (Author)

  11. Inhibition of the mixed leucocyte culture reaction by plasma from renal transplatn recipients.

    Science.gov (United States)

    Singal, D P; Younglai, E V; Naipaul, N

    1975-02-01

    Plasma samples from 28 renal allograft recipients were studied for the presence of alloantibodies blocking MLC response. MLC blocking antibodies were found in plasma samples obtained from 19 recipients, six with rejected allograft and 13 with long-term surviving transplant. In many instances, these antibodies were found in the absence of detectable lymphocytotoxic antibodies. The MLC blocking activity resides in IgG fractions of plasma samples and was decreased only after absorption with lymphocytes, no effect on blocking was observed by absorption with platelets and fibroblasts.

  12. NeuroAid (MLC601) versus piracetam in the recovery of post-infarct homonymous hemianopsia

    Institute of Scientific and Technical Information of China (English)

    Kavian Ghandehari; Zahra Izadi Mood; Saeed Ebrahimzadeh; David Picard; Yue Zhang

    2011-01-01

    In the clinic, the natural recovery rate of homonymous hemianopsia caused by occipital lobe infarction is low. At present, ideal therapeutic effects of piracetam for improving visual field defects following homonymous hemianopsia do not exist. The present randomized, controlled study compared the effects of NeuroAid (MLC601) versus piracetam for improving visual field defects in post-infarct homonymous hemianopsia patients matched for age and sex. After 3 months of treatment with NeuroAid (MLC601) or piracetam, visual field defects were significantly improved, compared with prior to treatment (P < 0.001). After treatment with MLC601, relative reduction of right and left visual field defects was 45% and 45.7%, respectively, while relative reduction after treatment with piracetam was 32.7% and 30.3%, respectively. These findings suggested that MLC601 was superior to piracetam for reducing visual field defects in homonymous hemianopsia patients.

  13. Real-time dynamic MLC tracking for inversely optimized arc radiotherapy

    DEFF Research Database (Denmark)

    Falk, Marianne; af Rosenschöld, Per Munck; Keall, Paul;

    2010-01-01

    Motion compensation with MLC tracking was tested for inversely optimized arc radiotherapy with special attention to the impact of the size of the target displacements and the angle of the leaf trajectory....

  14. Monte Carlo implementation, validation, and characterization of a 120 leaf MLC

    Energy Technology Data Exchange (ETDEWEB)

    Fix, Michael K.; Volken, Werner; Frei, Daniel; Frauchiger, Daniel; Born, Ernst J.; Manser, Peter [Division of Medical Radiation Physics, Inselspital and University of Bern, CH-3010 Bern (Switzerland)

    2011-10-15

    Purpose: Recently, the new high definition multileaf collimator (HD120 MLC) was commercialized by Varian Medical Systems providing high resolution in the center section of the treatment field. The aim of this work is to investigate the characteristics of the HD120 MLC using Monte Carlo (MC) methods. Methods: Based on the information of the manufacturer, the HD120 MLC was implemented into the already existing Swiss MC Plan (SMCP). The implementation has been configured by adjusting the physical density and the air gap between adjacent leaves in order to match transmission profile measurements for 6 and 15 MV beams of a Novalis TX. These measurements have been performed in water using gafchromic films and an ionization chamber at an SSD of 95 cm and a depth of 5 cm. The implementation was validated by comparing diamond measured and calculated penumbra values (80%-20%) for different field sizes and water depths. Additionally, measured and calculated dose distributions for a head and neck IMRT case using the DELTA{sup 4} phantom have been compared. The validated HD120 MLC implementation has been used for its physical characterization. For this purpose, phase space (PS) files have been generated below the fully closed multileaf collimator (MLC) of a 40 x 22 cm{sup 2} field size for 6 and 15 MV. The PS files have been analyzed in terms of energy spectra, mean energy, fluence, and energy fluence in the direction perpendicular to the MLC leaves and have been compared with the corresponding data using the well established Varian 80 leaf (MLC80) and Millennium M120 (M120 MLC) MLCs. Additionally, the impact of the tongue and groove design of the MLCs on dose has been characterized. Results: Calculated transmission values for the HD120 MLC are 1.25% and 1.34% in the central part of the field for the 6 and 15 MV beam, respectively. The corresponding ionization chamber measurements result in a transmission of 1.20% and 1.35%. Good agreement has been found for the comparison

  15. Fast motion-including dose error reconstruction for VMAT with and without MLC tracking

    DEFF Research Database (Denmark)

    Ravkilde, Thomas; Keall, Paul J.; Grau, Cai

    2014-01-01

    validate a simple model for fast motion-including dose error reconstruction applicable to intrafractional QA of MLC tracking treatments of moving targets. MLC tracking experiments were performed on a standard linear accelerator with prototype MLC tracking software guided by an electromagnetic transponder...... system. A three-axis motion stage reproduced eight representative tumour trajectories; four lung and four prostate. Low and high modulation 6 MV single-arc volumetric modulated arc therapy treatment plans were delivered for each trajectory with and without MLC tracking, as well as without motion...... for reference. Temporally resolved doses were measured during all treatments using a biplanar dosimeter. Offline, the dose delivered to each of 1069 diodes in the dosimeter was reconstructed with 500 ms temporal resolution by a motion-including pencil beam convolution algorithm developed in-house. The accuracy...

  16. The first clinical implementation of electromagnetic transponder-guided MLC tracking

    Energy Technology Data Exchange (ETDEWEB)

    Keall, Paul J., E-mail: paul.keall@sydney.edu.au; O’Brien, Ricky; Ng, Jin Aun [Radiation Physics Laboratory, Sydney Medical School, University of Sydney, NSW 2006 (Australia); Colvill, Emma [Radiation Physics Laboratory, Sydney Medical School, University of Sydney, NSW 2006, Australia and Northern Sydney Cancer Centre, Royal North Shore Hospital, Sydney, NSW 2065 (Australia); Poulsen, Per Rugaard [Department of Oncology, Aarhus University Hospital, Aarhus 8000, Denmark and Institute of Clinical Medicine, Aarhus University, Aarhus 8000 (Denmark); Eade, Thomas; Kneebone, Andrew; Booth, Jeremy T. [Northern Sydney Cancer Centre, Royal North Shore Hospital, Sydney, NSW 2065 (Australia)

    2014-02-15

    Purpose: We report on the clinical process, quality assurance, and geometric and dosimetric results of the first clinical implementation of electromagnetic transponder-guided MLC tracking which occurred on 28 November 2013 at the Northern Sydney Cancer Centre. Methods: An electromagnetic transponder-based positioning system (Calypso) was modified to send the target position output to in-house-developed MLC tracking code, which adjusts the leaf positions to optimally align the treatment beam with the real-time target position. Clinical process and quality assurance procedures were developed and performed. The first clinical implementation of electromagnetic transponder-guided MLC tracking was for a prostate cancer patient being treated with dual-arc VMAT (RapidArc). For the first fraction of the first patient treatment of electromagnetic transponder-guided MLC tracking we recorded the in-room time and transponder positions, and performed dose reconstruction to estimate the delivered dose and also the dose received had MLC tracking not been used. Results: The total in-room time was 21 min with 2 min of beam delivery. No additional time was needed for MLC tracking and there were no beam holds. The average prostate position from the initial setup was 1.2 mm, mostly an anterior shift. Dose reconstruction analysis of the delivered dose with MLC tracking showed similar isodose and target dose volume histograms to the planned treatment and a 4.6% increase in the fractional rectal V{sub 60}. Dose reconstruction without motion compensation showed a 30% increase in the fractional rectal V{sub 60} from that planned, even for the small motion. Conclusions: The real-time beam-target correction method, electromagnetic transponder-guided MLC tracking, has been translated to the clinic. This achievement represents a milestone in improving geometric and dosimetric accuracy, and by inference treatment outcomes, in cancer radiotherapy.

  17. Real-time dynamic MLC tracking for inversely optimized arc radiotherapy

    DEFF Research Database (Denmark)

    Falk, Marianne; af Rosenschöld, Per Munck; Keall, Paul

    2010-01-01

    Motion compensation with MLC tracking was tested for inversely optimized arc radiotherapy with special attention to the impact of the size of the target displacements and the angle of the leaf trajectory.......Motion compensation with MLC tracking was tested for inversely optimized arc radiotherapy with special attention to the impact of the size of the target displacements and the angle of the leaf trajectory....

  18. Partial transmission block production for real efficient method of block and MLC

    Energy Technology Data Exchange (ETDEWEB)

    Choi, Ji Min; Park, Ju Young; Ju, Sang Kyu; Park, Jong Ho [Dept. of Radiation Oncology, Samsung Medical Center, Sunkyunkwan University, Seoul (Korea, Republic of)

    2004-09-15

    The Vaginal, the urethra, the vulva and anal cancer avoid the many dose to femur head and the additional treatment is necessary in inguinal LN. The partial transmission block to use inguinal LN addition there is to a method which it treats and produce partial transmission block a method and the MLC which to it analyzes. The Inguinal the LN treatment patient partial transmission it used block and the MLC in the object and with solid water phantom with the patient it reappeared the same depth. In order to analyze the error of the junction the EDR2 (Extended dose range, the Kodak and the U.S) it used the Film and it got film scanner it got the beam profile. The partial transmission block and the MLC bias characteristic, accuracy and stability of production for, it shared at hour and comparison it analyzed. The partial the transmission block compares in the MLC and the block production is difficult and production hour also above 1 hours. The custom the block the place where it revises the error of the junction is a difficult problem. If use of the MLC the fabrication will be break and only the periodical calibration of the MLC it will do and it will be able to use easily. The Inguinal there is to LN treatment and partial transmission block and the MLC there is efficiency of each one but there is a place where the junction of block for partial transmission block the production hour is caught long and it fixes and a point where the control of the block is difficult. Like this problem it transfers with the MLC and if it treats, it means the effective treatment will be possible.

  19. Inhibition of nucleic acid synthesis in P388 lymphocytic leukemia cells in culture by cis-platinum derivatives.

    Science.gov (United States)

    Oswald, C B; Chaney, S G; Hall, I H

    1990-01-01

    Cis-diaminedichloroplatinum(II) [cDDP] and three related derivatives Pt(mal)(NH3)2, PtCl2(dach) and Pt(mal) (dach) have been observed to possess cytotoxicity against the growth of P388 lymphocytic leukemia cells. DNA synthesis in P388 cells was inhibited by the agents in a manner which was consistent with their ED50 values for cytotoxicity. When P388 cells were treated with these platinum complexes in vitro at doses which caused more than 80% inhibition of DNA synthesis, no significant inhibition was observed for thymidine, kinase, thymidine monophosphate kinase, carbamoyl phosphate synthetase, or aspartate transcarbamoylase activities. Thus, there was no evidence that these agents inhibited de novo purine, pyrmidine, or deoxynucleotide synthesis. All of the agents did inhibit the nuclear DNA polymerase activity, but the extent of inhibition was 20% or less at doses which caused greater than 70% inhibition of DNA synthesis. Thus, the inhibition of DNA synthesis appeared to be due to cisplatinum(II) drug binding to the DNA bases. This was estimated to be 1 atom of platinum per 1500-3000 DNA base pairs which is consistent with other studies. The platinum complexes with chloro leaving ligands caused considerable DNA strand scission by 24 h at 10 times the ED50 dose, most likely a measure of impending cell death. In contrast, the platinum complexes with malonato leaving ligands did not cause significant strand scission by 24 h at similar doses. They also exhibited a significant delay in the inhibition of DNA synthesis. These data were interpreted as resulting from slower monoadduct to diadduct conversion, but it is not possible to eliminate the possibility of a different mode of interaction with DNA or a different mechanism of cytotoxicity for the malonato compounds.

  20. Yeast myosin light chain, Mlc1p, interacts with both IQGAP and class II myosin to effect cytokinesis.

    Science.gov (United States)

    Boyne, J R; Yosuf, H M; Bieganowski, P; Brenner, C; Price, C

    2000-12-01

    MLC1 (myosin light chain) acts as a dosage suppressor of a temperature sensitive mutation in the gene encoding the S. cerevisiae IQGAP protein. Both proteins localize to the bud neck in mitosis although Mlc1p localisation precedes Iqg1p. Mlc1p is also found at the incipient bud site in G(1) and the growing bud tip during S and G(2) phases of the cell cycle. A dominant negative GST-Mlc1p fusion protein specifically blocks cytokinesis and prevents Iqg1p localisation to the bud neck, as does depletion of Mlc1p. These data support a direct interaction between the two proteins and immunoprecipitation experiments confirm this prediction. Mlc1p is also shown to interact with the class II conventional myosin (Myo1p). All three proteins form a complex, however, the interaction between Mlc1p and Iqg1p can be separated from the Mlc1p/Myo1p interaction. Mlc1p localisation and maintenance at the bud neck is independent of actin, Myo1p and Iqg1p. It is proposed that Mlc1p therefore functions to recruit Iqg1p and in turn actin to the actomyosin ring and that it is also required for Myo1p function during ring contraction.

  1. Detecting MLC errors in stereotactic radiotherapy plans with a liquid filled ionization chamber array.

    Science.gov (United States)

    O'Connor, Patrick; Seshadri, Venkatakrisnan; Charles, Paul

    2016-03-01

    Quality assurance of stereotactic radiotherapy demands the use of equipment with the highest resolution and sensitivity available. This study examines the sensitivity of a commercially available liquid-filled ionization chamber array--the Octavius 1000 SRS (PTW, Frieburg, Germany) for detecting small (sub-millimetre) multi-leaf collimator (MLC) alignment errors in static square fields (side length 16-40 mm). Furthermore, the effectiveness of detecting small MLC errors in clinical stereotactic radiotherapy patient plans using the device was also evaluated. The commonly used gamma pass rate metric (of the measurements compared with treatment planning system generated results) was used. The gamma pass rates were then evaluated as a function of MLC position error (MLC error size 0.1-2.5 mm). The detector array exhibited a drop in pass rate between plans without error and those which had MLC errors induced. For example a drop in pass rate of 4.5% (gamma criteria 3%, 1 mm) was observed when a 0.8 mm error was introduced into a 16 mm square field. Furthermore the drop in pass rate increased as the MLC position error increased. This study showed that the Octavius 1000 SRS array could be a useful tool for applications requiring the detection of small geometric delivery uncertainties.

  2. Detailed analysis of latencies in image-based dynamic MLC tracking

    Energy Technology Data Exchange (ETDEWEB)

    Poulsen, Per Rugaard; Cho, Byungchul; Sawant, Amit; Ruan, Dan; Keall, Paul J. [Department of Radiation Oncology, Stanford University, Stanford, California 94305 and Department of Oncology and Department of Medical Physics, Aarhus University Hospital, 8000 Aarhus (Denmark); Department of Radiation Oncology, Stanford University, Stanford, California 94305 and Department of Radiation Oncology, Asan Medical Center, Seoul 138-736 (Korea, Republic of); Department of Radiation Oncology, Stanford University, Stanford, California 94305 (United States)

    2010-09-15

    Purpose: Previous measurements of the accuracy of image-based real-time dynamic multileaf collimator (DMLC) tracking show that the major contributor to errors is latency, i.e., the delay between target motion and MLC response. Therefore the purpose of this work was to develop a method for detailed analysis of latency contributions during image-based DMLC tracking. Methods: A prototype DMLC tracking system integrated with a linear accelerator was used for tracking a phantom with an embedded fiducial marker during treatment delivery. The phantom performed a sinusoidal motion. Real-time target localization was based on x-ray images acquired either with a portal imager or a kV imager mounted orthogonal to the treatment beam. Each image was stored in a file on the imaging workstation. A marker segmentation program opened the image file, determined the marker position in the image, and transferred it to the DMLC tracking program. This program estimated the three-dimensional target position by a single-imager method and adjusted the MLC aperture to the target position. Imaging intervals {Delta}T{sub image} from 150 to 1000 ms were investigated for both kV and MV imaging. After the experiments, the recorded images were synchronized with MLC log files generated by the MLC controller and tracking log files generated by the tracking program. This synchronization allowed temporal analysis of the information flow for each individual image from acquisition to completed MLC adjustment. The synchronization also allowed investigation of the MLC adjustment dynamics on a considerably finer time scale than the 50 ms time resolution of the MLC log files. Results: For {Delta}T{sub image}=150 ms, the total time from image acquisition to completed MLC adjustment was 380{+-}9 ms for MV and 420{+-}12 ms for kV images. The main part of this time was from image acquisition to completed image file writing (272 ms for MV and 309 ms for kV). Image file opening (38 ms), marker segmentation (4 ms

  3. Studies of the response in mixed leukocyte culture of cells from patients with aplastic anemia to cells from HLA-identical siblings.

    Science.gov (United States)

    Mickelson, E M; Clift, R A; Fefer, A; Storb, R; Thomas, E D; Warren, R P; Hansen, J A

    1981-08-01

    We have studied the mixed leukocyte culture (MLC) reactions of 64 patients with severe aplastic anemia. Their peripheral blood mononuclear cells showed an increased relative response (RR) to cells from HLA-identical siblings as compared to cells from normal HLA-identical siblings, confirming the results reported in an earlier series of 34 patients. Elevated RRs were associated with patient antidonor lymphocyte antibodies as detected by the antibody-dependent cell-mediated cytotoxicity assay, but were not associated with antidonor complement-dependent cytotoxic antibodies or with antidonor cytotoxic T lymphocytes. Among 98 patients receiving marrow grafts from HLA-identical sibling donors, those with elevated RRs before transplantation were more apt to reject the transplant than those without (P less than 0.0001). There was no elevation of the RR in 10 untransfused patients, although positive RRs were noted in 2 patients within 12 to 24 hr of their first transfusions. Five patients with identical twins were also tested, and elevated RRs were noted in three. Although blood transfusion appears to be responsible for the increased RRs observed in some aplastic patients, genetic differences between donor and recipient were not always necessary for eliciting an increased MLC response, suggesting that mechanisms other than alloimmunization are involved.

  4. Megalencephalic leukoencephalopathy with subcortical cysts protein-1 modulates endosomal pH and protein trafficking in astrocytes: relevance to MLC disease pathogenesis.

    Science.gov (United States)

    Brignone, Maria S; Lanciotti, Angela; Visentin, Sergio; De Nuccio, Chiara; Molinari, Paola; Camerini, Serena; Diociaiuti, Marco; Petrini, Stefania; Minnone, Gaetana; Crescenzi, Marco; Laudiero, Luisa Bracci; Bertini, Enrico; Petrucci, Tamara C; Ambrosini, Elena

    2014-06-01

    Megalencephalic leukoencephalopathy with subcortical cysts (MLC) is a rare leukodystrophy caused by mutations in the gene encoding MLC1, a membrane protein mainly expressed in astrocytes in the central nervous system. Although MLC1 function is unknown, evidence is emerging that it may regulate ion fluxes. Using biochemical and proteomic approaches to identify MLC1 interactors and elucidate MLC1 function we found that MLC1 interacts with the vacuolar ATPase (V-ATPase), the proton pump that regulates endosomal acidity. Because we previously showed that in intracellular organelles MLC1 directly binds Na, K-ATPase, which controls endosomal pH, we studied MLC1 endosomal localization and trafficking and MLC1 effects on endosomal acidity and function using human astrocytoma cells overexpressing wild-type (WT) MLC1 or MLC1 carrying pathological mutations. We found that WT MLC1 is abundantly expressed in early (EEA1(+), Rab5(+)) and recycling (Rab11(+)) endosomes and uses the latter compartment to traffic to the plasma membrane during hyposmotic stress. We also showed that WT MLC1 limits early endosomal acidification and influences protein trafficking in astrocytoma cells by stimulating protein recycling, as revealed by FITC-dextran measurement of endosomal pH and transferrin protein recycling assay, respectively. WT MLC1 also favors recycling to the plasma-membrane of the TRPV4 cation channel which cooperates with MLC1 to activate calcium influx in astrocytes during hyposmotic stress. Although MLC disease-causing mutations differentially affect MLC1 localization and trafficking, all the mutated proteins fail to influence endosomal pH and protein recycling. This study demonstrates that MLC1 modulates endosomal pH and protein trafficking suggesting that alteration of these processes contributes to MLC pathogenesis. Copyright © 2014. Published by Elsevier Inc.

  5. The use of FISH-comet to detect c-Myc and TP 53 damage in extended-term lymphocyte cultures treated with terbuthylazine and carbofuran.

    Science.gov (United States)

    Mladinic, Marin; Zeljezic, Davor; Shaposhnikov, Sergey A; Collins, Andrew R

    2012-05-20

    Terbuthylazine and carbofuran are suspected to cause non-Hodgkin's lymphoma and lung cancer. We evaluated the effects of prolonged exposure to low concentrations on primary DNA damage by comet assay, and on the structural integrity of c-Myc and TP 53 genes by FISH-comet. Another novelty in studying these pesticides' genotoxicity is the use of 14-day extended-term human lymphocyte cultures. Concentrations corresponded to values of ADI and OEL: for terbuthylazine 0.58 ng/ml and 8 ng/ml; for carbofuran 8 ng/ml and 21.6 ng/ml, respectively. A possible effect of metabolic activation (S9) was also considered. Carbofuran treatment induced a significant migration of DNA into the tail in a concentration-dependent manner, while for terbuthylazine the effect was significant only at the higher concentration. Terbuthylazine caused migration of both c-Myc signals into the comet tail. A significant occurrence of TP 53 signals in the tail was observed at 8 ng/ml. Prolonged carbofuran treatment significantly elevated the migration of a single c-Myc signal into the tail in a concentration-dependent manner. With S9, distribution of signals shifted toward increased presence of both signals in tail. Our results showed impaired structural integrity of c-Myc and TP 53 due to prolonged exposure to terbuthylazine and carbofuran.

  6. Could Intrathymic Injection of Myelin Basic Protein Suppress Inflammatory Response After Co-culture of T Lymphocytes and BV-2 Microglia Cells?

    Institute of Scientific and Technical Information of China (English)

    Zhan-Qun Cui; Bao-Long Liu; Qiao-Li Wu; Ying Cai; Wei-Jia Fan; Ming-Chao Zhang; Wei-Liang Ding

    2016-01-01

    Background:The interaction between activated microglia and T lymphocytes can yield abundant pro-inflammatory cytokines.Our previous study proved that thymus immune tolerance could alleviate the inflammatory response.This study aimed to investigate whether intrathymic injection of myelin basic protein (MBP) in mice could suppress the inflammatory response after co-culture ofT lymphocytes and BV-2 microglia cells.Methods:Totally,72 male C57BL/6 mice were randomly assigned to three groups (n =24 in each):Group A:intrathymic injection of 100 μl MBP (1 mg/ml);Group B:intrathymic injection of 100 μl phosphate-buffered saline (PBS);and Group C:sham operation group.Every eight mice in each group were sacrificed to obtain the spleen at postoperative days 3,7,and 14,respectively.T lymphocytes those were extracted and purified from the spleens were then co-cultured with activated BV-2 microglia cells at a proportion of 1:2 in the medium containing MBP for 3 days.After identified the T lymphocytes by CD3,surface antigens of T lymphocytes (CD4,CD8,CD152,and CD154) and BV-2 microglia cells (CD45 and CD54) were detected by flow cytometry.The expressions of pro-inflammatory factors ofBV-2 microglia cells (interleukin [IL]-1β,tumor necrosis factor-α [TNF-α],and inducible nitric oxide synthase [iNOS]) were detected by quantitative real-time polymerase chain reaction (PCR).One-way analysis of variance (ANOVA) and the least significant difference test were used for data analysis.Results:The levels of CD152 in Group A showed an upward trend from the 3rd to 7th day,with a downward trend from the 7th to 14th day (20.12 ± 0.71%,30.71 ± 1.14%,13.50 ± 0.71% at postoperative days 3,7,and 14,respectively,P < 0.05).The levels of CD154 in Group A showed a downward trend from the 3rd to 7th day,with an upward trend from the 7th to 14th day (10.00 ± 0.23%,5.28 ± 0.69%,14.67 ± 2.71% at postoperative days 3,7,and 14,respectively,P < 0.05).The ratio ofCD4+/CD8 + T in Group

  7. Automatic detection of MLC relative position errors for VMAT using the EPID-based picket fence test

    Science.gov (United States)

    Christophides, Damianos; Davies, Alex; Fleckney, Mark

    2016-12-01

    Multi-leaf collimators (MLCs) ensure the accurate delivery of treatments requiring complex beam fluences like intensity modulated radiotherapy and volumetric modulated arc therapy. The purpose of this work is to automate the detection of MLC relative position errors  ⩾0.5 mm using electronic portal imaging device-based picket fence tests and compare the results to the qualitative assessment currently in use. Picket fence tests with and without intentional MLC errors were measured weekly on three Varian linacs. The picket fence images analysed covered a time period ranging between 14-20 months depending on the linac. An algorithm was developed that calculated the MLC error for each leaf-pair present in the picket fence images. The baseline error distributions of each linac were characterised for an initial period of 6 months and compared with the intentional MLC errors using statistical metrics. The distributions of median and one-sample Kolmogorov-Smirnov test p-value exhibited no overlap between baseline and intentional errors and were used retrospectively to automatically detect MLC errors in routine clinical practice. Agreement was found between the MLC errors detected by the automatic method and the fault reports during clinical use, as well as interventions for MLC repair and calibration. In conclusion the method presented provides for full automation of MLC quality assurance, based on individual linac performance characteristics. The use of the automatic method has been shown to provide early warning for MLC errors that resulted in clinical downtime.

  8. Optimisation of the CT h4S bioassay for detection of human interleukin-4 secreted by mononuclear cells stimulated by phytohaemaglutinin or by human leukocyte antigen mismatched mixed lymphocyte culture

    DEFF Research Database (Denmark)

    Petersen, Søren Lykke; Russell, Charlotte Astrid; Bendtzen, Klaus;

    2002-01-01

    bioassay with regards to specificity, sensitivity, detection limit, and reproducibility. We have found the optimal assay conditions to be 1 x 10 (4) CT.h4S cells/well deprived of IL-4 for 24 h and preincubated for 7 h followed by 18 h of incubation with tritiated methyl-thymidine. In this setting the CT.h4......S bioassay detects 5 pg/ml of human recombinant IL-4 with no detection of IL-2 in concentrations below 500 pg/ml. We have found 72 h of culture optimal for detection of IL-2 and IL-4 produced by human mononuclear cells (MNC) in response to stimulation with phytohaemaglutinin and for detection of IL...... of IL-4 detection was not due to high amounts of soluble IL-4 receptor. With the use of 1x10(6) responder cells/well in HLA-mismatched MLC, we found limited IL-4 accumulation still increasing at day 12. We conclude that the CT.h4S bioassay is a reliable and specific method for quantification of IL-4...

  9. Dose domain regularization of MLC leaf patterns for highly complex IMRT plans

    Energy Technology Data Exchange (ETDEWEB)

    Nguyen, Dan; Yu, Victoria Y.; Ruan, Dan; Cao, Minsong; Low, Daniel A.; Sheng, Ke, E-mail: ksheng@mednet.ucla.edu [Department of Radiation Oncology, University of California Los Angeles, Los Angeles, California 90095 (United States); O’Connor, Daniel [Department of Mathematics, University of California Los Angeles, Los Angeles, California 90095 (United States)

    2015-04-15

    Purpose: The advent of automated beam orientation and fluence optimization enables more complex intensity modulated radiation therapy (IMRT) planning using an increasing number of fields to exploit the expanded solution space. This has created a challenge in converting complex fluences to robust multileaf collimator (MLC) segments for delivery. A novel method to regularize the fluence map and simplify MLC segments is introduced to maximize delivery efficiency, accuracy, and plan quality. Methods: In this work, we implemented a novel approach to regularize optimized fluences in the dose domain. The treatment planning problem was formulated in an optimization framework to minimize the segmentation-induced dose distribution degradation subject to a total variation regularization to encourage piecewise smoothness in fluence maps. The optimization problem was solved using a first-order primal-dual algorithm known as the Chambolle-Pock algorithm. Plans for 2 GBM, 2 head and neck, and 2 lung patients were created using 20 automatically selected and optimized noncoplanar beams. The fluence was first regularized using Chambolle-Pock and then stratified into equal steps, and the MLC segments were calculated using a previously described level reducing method. Isolated apertures with sizes smaller than preset thresholds of 1–3 bixels, which are square units of an IMRT fluence map from MLC discretization, were removed from the MLC segments. Performance of the dose domain regularized (DDR) fluences was compared to direct stratification and direct MLC segmentation (DMS) of the fluences using level reduction without dose domain fluence regularization. Results: For all six cases, the DDR method increased the average planning target volume dose homogeneity (D95/D5) from 0.814 to 0.878 while maintaining equivalent dose to organs at risk (OARs). Regularized fluences were more robust to MLC sequencing, particularly to the stratification and small aperture removal. The maximum and

  10. Evaluation of dosimetric effect caused by slowing with multi-leaf collimator (MLC) leaves for volumetric modulated arc therapy (VMAT).

    Science.gov (United States)

    Xu, Zhengzheng; Wang, Iris Z; Kumaraswamy, Lalith K; Podgorsak, Matthew B

    2016-03-01

    This study is to report 1) the sensitivity of intensity modulated radiation therapy (IMRT) QA method for clinical volumetric modulated arc therapy (VMAT) plans with multi-leaf collimator (MLC) leaf errors that will not trigger MLC interlock during beam delivery; 2) the effect of non-beam-hold MLC leaf errors on the quality of VMAT plan dose delivery. Eleven VMAT plans were selected and modified using an in-house developed software. For each control point of a VMAT arc, MLC leaves with the highest speed (1.87-1.95 cm/s) were set to move at the maximal allowable speed (2.3 cm/s), which resulted in a leaf position difference of less than 2 mm. The modified plans were considered as 'standard' plans, and the original plans were treated as the 'slowing MLC' plans for simulating 'standard' plans with leaves moving at relatively lower speed. The measurement of each 'slowing MLC' plan using MapCHECK®2 was compared with calculated planar dose of the 'standard' plan with respect to absolute dose Van Dyk distance-to-agreement (DTA) comparisons using 3%/3 mm and 2%/2 mm criteria. All 'slowing MLC' plans passed the 90% pass rate threshold using 3%/3 mm criteria while one brain and three anal VMAT cases were below 90% with 2%/2 mm criteria. For ten out of eleven cases, DVH comparisons between 'standard' and 'slowing MLC' plans demonstrated minimal dosimetric changes in targets and organs-at-risk. For highly modulated VMAT plans, pass rate threshold (90%) using 3%/3mm criteria is not sensitive in detecting MLC leaf errors that will not trigger the MLC leaf interlock. However, the consequential effects of non-beam hold MLC errors on target and OAR doses are negligible, which supports the reliability of current patient-specific IMRT quality assurance (QA) method for VMAT plans.

  11. In vitro and in vivo analyses of a genetically—restricted antigen specific factor from mixed cell cultures of macrophage,T and B lymphocytes

    Institute of Scientific and Technical Information of China (English)

    CHAURMW; LAUASK

    1990-01-01

    An immunostimulatory factor was identified to be secreted by antigen-pulsed macrophages.This factor was able to induce the generation of antigen specific T helper lymphocytes in vitro as well as in vivo.Further in vitro experiments testing for the genetic restriction of this factor indicated that it is a geneticallyrestricted antigen specific factor (ASF).The Cunningham plaque assay was used to quantify the generation of T helper lymphocytes by measuring the number of plaque forming cells after sequential incubations of antigen-qulsed macrophages with T lymphocytes,and then spleen cells,and finally the TNP-coated sheep red blood cells.

  12. Fast motion-including dose error reconstruction for VMAT with and without MLC tracking

    DEFF Research Database (Denmark)

    Ravkilde, Thomas; Keall, Paul J.; Grau, Cai

    2014-01-01

    of the algorithm for reconstruction of dose and motion-induced dose errors throughout the tracking and non-tracking beam deliveries was quantified. Doses were reconstructed with a mean dose difference relative to the measurements of -0.5% (5.5% standard deviation) for cumulative dose. More importantly, the root......-mean-square deviation between reconstructed and measured motion-induced 3%/3 mm γ failure rates (dose error) was 2.6%. The mean computation time for each calculation of dose and dose error was 295 ms. The motion-including dose reconstruction allows accurate temporal and spatial pinpointing of errors in absorbed dose...... validate a simple model for fast motion-including dose error reconstruction applicable to intrafractional QA of MLC tracking treatments of moving targets. MLC tracking experiments were performed on a standard linear accelerator with prototype MLC tracking software guided by an electromagnetic transponder...

  13. Multiple linear combination (MLC) regression tests for common variants adapted to linkage disequilibrium structure

    Science.gov (United States)

    Yoo, Yun Joo; Sun, Lei; Poirier, Julia G.; Paterson, Andrew D.

    2016-01-01

    ABSTRACT By jointly analyzing multiple variants within a gene, instead of one at a time, gene‐based multiple regression can improve power, robustness, and interpretation in genetic association analysis. We investigate multiple linear combination (MLC) test statistics for analysis of common variants under realistic trait models with linkage disequilibrium (LD) based on HapMap Asian haplotypes. MLC is a directional test that exploits LD structure in a gene to construct clusters of closely correlated variants recoded such that the majority of pairwise correlations are positive. It combines variant effects within the same cluster linearly, and aggregates cluster‐specific effects in a quadratic sum of squares and cross‐products, producing a test statistic with reduced degrees of freedom (df) equal to the number of clusters. By simulation studies of 1000 genes from across the genome, we demonstrate that MLC is a well‐powered and robust choice among existing methods across a broad range of gene structures. Compared to minimum P‐value, variance‐component, and principal‐component methods, the mean power of MLC is never much lower than that of other methods, and can be higher, particularly with multiple causal variants. Moreover, the variation in gene‐specific MLC test size and power across 1000 genes is less than that of other methods, suggesting it is a complementary approach for discovery in genome‐wide analysis. The cluster construction of the MLC test statistics helps reveal within‐gene LD structure, allowing interpretation of clustered variants as haplotypic effects, while multiple regression helps to distinguish direct and indirect associations. PMID:27885705

  14. 同种异体淋巴细胞与黑素细胞混合培养的实验研究%Allogeneic Mixed Melanocyte-Lymphocyte Culture: An Experimental Study

    Institute of Scientific and Technical Information of China (English)

    项蕾红; 郑志忠; 祝绿川; 廖康煌; 傅雯雯

    2001-01-01

    Objective To investigate the melanocyte-lymphocyte reacion when the allogeneic lymphocytes are cultured with normal melanocytes. Methods 3H - thymidine was incorporated into the mixed culture and the transformation and proliferation rates of lymphocytes were detected by liquid scintillation counting and expressed as cpm. Electron microscopy was used to observe the ultrastructure of melanocytes after mixed culture.Results The results of lymphocyte proliferation were expressed by the stimulation indexes. The stimulation indexes in active vitiligo group was significantly different from that in stable vitiligo group and normal controls. The stimulation indexes of the melanocyte-stimulated group was significantly different from that of ConA-stimulated group. In the mixed melanocyte-lymphocyte reaction, the allogeneic lymphocytes had little effect on the melanocytes. The ultrastructure of the melanocytes in the mixed culture showed normal morphology and normal function of synthesis of melanin. Conclusion As a specific antigen in mixed melanocyte-lymphocyte reaction, melanocyte has a weak effect on the lymphocytes. The melanocytes from stable stage vitiligo patients seem more suitble to be allografted.%目的黑素细胞与同种异体淋巴细胞混合培养,从体外实验来研究排斥反应的可能性。方法采用3H-dTR掺入同位素液态闪烁计数法测定淋巴细胞的转化增殖率,并用透射电镜观察混合培养后黑素细胞的超微结构。结果黑素细胞对同种异体淋巴细胞的转化增殖率与ConA刺激淋巴细胞转化增殖的阳性对照比较,结果显示黑素细胞的促淋巴细胞转化增殖的特异性抗原作用较弱。进一步比较黑素细胞对不同病期白癜风患者淋巴细胞的影响,发现黑素细胞对活动期白癜风患者淋巴细胞的刺激作用相对较强,而稳定期患者和正常人对照组结果差异无显著性。同时电镜观察结果显示同种异体的淋巴细胞对黑

  15. SU-C-BRB-02: Symmetric and Asymmetric MLC Based Lung Shielding and Dose Optimization During Translating Bed TBI

    Energy Technology Data Exchange (ETDEWEB)

    Ahmed, S; Kakakhel, MB [Pakistan Institute of Engineering & Applied Sciences (PIEAS), Islamabad (Pakistan); Ahmed, SBS; Hussain, A [Aga Khan University Hospital (AKUH), Karachi (Pakistan)

    2015-06-15

    Purpose: The primary aim was to introduce a dose optimization method for translating bed total body irradiation technique that ensures lung shielding dynamically. Symmetric and asymmetric dynamic MLC apertures were employed for this purpose. Methods: The MLC aperture sizes were defined based on the radiological depth values along the divergent ray lines passing through the individual CT slices. Based on these RD values, asymmetrically shaped MLC apertures were defined every 9 mm of the phantom in superior-inferior direction. Individual MLC files were created with MATLAB™ and were imported into Eclipse™ treatment planning system for dose calculations. Lungs can be shielded to an optimum level by reducing the MLC aperture width over the lungs. The process was repeated with symmetrically shaped apertures. Results: Dose-volume histogram (DVH) analysis shows that the asymmetric MLC based technique provides better dose coverage to the body and optimum shielding of the lungs compared to symmetrically shaped beam apertures. Midline dose homogeneity is within ±3% with asymmetric MLC apertures whereas it remains within ±4.5% with symmetric ones (except head region where it drops down to −7%). The substantial over and under dosage of ±5% at tissue interfaces has been reduced to ±2% with asymmetric MLC technique. Lungs dose can be reduced to any desired limit. In this experiment lungs dose was reduced to 80% of the prescribed dose, as was desired. Conclusion: The novel asymmetric MLC based technique assures optimum shielding of OARs (e.g. lungs) and better 3-D dose homogeneity and body-dose coverage in comparison with the symmetric MLC aperture optimization. The authors acknowledge the financial and infrastructural support provided by Pakistan Institute of Engineering & Applied Sciences (PIEAS), Islamabad and Aga Khan University Hospital (AKUH), Karachi during the course of this research project. Authors have no conflict of interest with any national / international

  16. Ivabradine Reduces Chemokine-Induced CD4-Positive Lymphocyte Migration

    Directory of Open Access Journals (Sweden)

    Thomas Walcher

    2010-01-01

    Full Text Available Aims. Migration of CD4-positive lymphocytes into the vessel wall is a critical step in atherogenesis. Recent data suggest that ivabradine, a selective I(f-channel blocker, reduces atherosclerotic plaque formation in apolipoprotein E-deficient mice, hitherto nothing is known about the mechanism by which ivabradine modulates plaque formation. Therefore, the present study investigated whether ivabradine regulates chemokine-induced migration of lymphocytes. Methods and results. Stimulation of CD4-positive lymphocytes with SDF-1 leads to a 2.0±0.1 fold increase in cell migration (P<.01; n=7. Pretreatment of cells with ivabradine reduces this effect to a maximal 1.2±0.1 fold induction at 0.1 µmol/L ivabradine (P<.01 compared to SDF-1-treated cells, n=7. The effect of ivabradine on CD4-positive lymphocyte migration was mediated through an early inhibition of chemokine-induced PI-3 kinase activity as determined by PI-3 kinase activity assays. Downstream, ivabradine inhibits activation of the small GTPase Rac and phosphorylation of the Myosin Light Chain (MLC. Moreover, ivabradine treatment reduces f-actin formation as well as ICAM3 translocation to the uropod of the cell, thus interfering with two important steps in T cell migration. Conclusion. Ivabradine inhibits chemokine-induced migration of CD4-positive lymphocytes. Given the crucial importance of chemokine-induced T-cell migration in early atherogenesis, ivabradine may be a promising tool to modulate this effect.

  17. Independent dose calculation of the Tps Iplan in radiotherapy conformed with MLC; Calculo independiente de dosis del TPS Iplan en radioterapia conformada con MLC

    Energy Technology Data Exchange (ETDEWEB)

    Adrada, A.; Tello, Z.; Medina, L.; Garrigo, E.; Venencia, D., E-mail: jorge.alberto.adrada@gmail.com [Instituto Privado de Radioterapia, Obispo Oro 423, X5000BFI Cordoba (Argentina)

    2014-08-15

    The systems utilization of independent dose calculation in three dimensional-Conformal Radiation Therapy (3D-Crt) treatments allows a direct verification of the treatments times. The utilization of these systems allows diminishing the probability of errors occurrence generated by the treatment planning system (Tps), allowing a detailed analysis of the dose to delivering and review of the normalization point (Np) or prescription. The independent dose calculation is realized across the knowledge of dosimetric parameters of the treatment machine and particular characteristics of every individual field. The aim of this work is develops a calculation system of punctual doses for isocentric fields conformed with multi-leaf collimation systems (MLC), where the dose calculation is in conformity with the suggested ones by ICRU Report No. 42, 1987. Calculation software was realized in C ++ under a free platform of programming (Code::Blocks). The system uses files in format Rtp, exported from the Tps to systems of record and verification (Lantis). This file contains detailed information of the dose, Um, position of the MLC sheets and collimators for every field of treatment. The size of equivalent field is obtained from the positions of every sheet; the effective depth of calculation can be introduced from the dosimetric report of the Tps or automatically from the DFS of the field. The 3D coordinates of the isocenter and the Np for the treatment plan must be introduced manually. From this information the system looks the dosimetric parameters and calculates the Um. The calculations were realized in two accelerators a NOVALIS Tx (Varian) with 120 sheets of high definition (hd-MLC) and a PRIMUS Optifocus (Siemens) with 82 sheets. 705 patients were analyzed for a total of 1082, in plans made for both equipment s, the average uncertainty with regard to the calculation of the Tps is-0.43% ± 2.42% in a range between [-7.90 %, 7.50 %]. The major uncertainty was in Np near of the

  18. [原著]Prethymic Nylon Wool-Passed Bone Marrow Cells Can Make Distinction between Self and Non-Self X-Chromosome-Linked Gene Products (Xir Antigens) on the Stimulator Cells, Resulting in Regulation of the Generation of Cytotoxic T Lymphocytes in Mixed Lymphocyte Cultures

    OpenAIRE

    Higa, Moritake; Tanabe, MasaoJ; Department of Bacteriology, Faculty of, University of the Ryukyus, Okinawa, Japan; Research Institute of Comprehensive Medicine, School of Medicine, University of the Ryukyus, Okinawa, Japan

    1994-01-01

    We have previously reported that nylon wool-passed bone marrow cells treated with anti-Thy.1 antibody and complement (Thy.1 NW-BM cells) had helper-like activity which could augment the generation of cytotoxic T lymphocytes (CTL). In this study, we determined the antigens to which these NW-BM cells responded and recognized. When a few responder lymph node (LN) cells and an excess of NW-BM responder cells from BIOBR (H-2^k B10 background) mice were cultured with stimulator spleen cells from ei...

  19. 6 MV dosimetric characterization of the 160 MLC, the new Siemens multileaf collimator.

    Science.gov (United States)

    Tacke, Martin B; Nill, Simeon; Häring, Peter; Oelfke, Uwe

    2008-05-01

    New technical developments constantly aim at improving the outcome of radiation therapy. With the use of a computer-controlled multileaf collimator (MLC), the quality of the treatment and the efficiency in patient throughput is significantly increased. New MLC designs aim to further enhance the advantages. In this article, we present the first detailed experimental investigation of the new 160 MLC, Siemens Medical Solutions. The assessment included the experimental investigation of typical MLC characteristics such as leakage, tongue-and-groove effect, penumbra, leaf speed, and leaf positioning accuracy with a 6 MV treatment beam. The leakage is remarkably low with an average of 0.37% due to a new design principle of slightly tilted leaves instead of the common tongue-and-groove design. But due to the tilt, the triangular tongue-and-groove effect occurs. Its magnitude of approximately 19% is similar to the dose defect measured for MLCs with the common tongue-and-groove design. The average longitudinal penumbra measured at depth d(max) = 15 mm with standard 100 x 100 mm2 fields is 4.1 +/- 0.5 mm for the central range and increases to 4.9 +/- 1.3 mm for the entire field range of 400 x 400 mm2. The increase is partly due to the single-focusing design and the large distance between the MLC and the isocenter enabling a large patient clearance. Regarding the leaf speed, different velocity tests were performed. The positions of the moving leaves were continuously recorded with the kilovoltage-imaging panel. The maximum leaf velocities measured were 42.9 +/- 0.6 mm/s. In addition, several typical intensity-modulated radiation therapy treatments were performed and the delivery times compared to the Siemens OPTIFOCUS MLC. An average decrease of 11% in delivery time was observed. The experimental results presented in this article indicate that the dosimetric characteristics of the 160 MLC are capable of improving the quality of dose delivery with respect to precision and dose

  20. Unusual 5'-regulatory structure and regulation of the murine Mlc1 gene: Lack of promoter-specific functional elements

    Directory of Open Access Journals (Sweden)

    Darja Henseler

    2011-10-01

    Full Text Available Normal 0 21 false false false MicrosoftInternetExplorer4 The MLC1 gene is involved in an autosomal recessive neurological disorder, megalencephalic leucoencephalopathy with subcortical cysts (MLC, which is characterized by macrocephaly during the first year of life and swollen white matter (leucoencephaly. Variants of MLC1 have also been associated with psychiatric disorders such as schizophrenia, major depression and bipolar disorder. Currently, little is known about the encoded protein (MLC1. Judging from its similarity to other known proteins, it may serve as a trans-membrane transporter. However, the function of the encoded protein and its gene regulation has not been investigated successfully so far. We investigated the 5’ region of the murine Mlc1 with respect to regulatory elements for gene expression. A promoter search and an in silico analysis were conducted. Luciferase reporter gene constructs with potential promoter regions were created to study promoter activity in vitro. We found two alternative first exons for the murine Mlc1 but were not able to detect any promoter activity for the investigated reporter gene constructs in different cell lines, thus pointing to the presence of essential cis-acting elements far outside of the region. In silico analysis indicated an uncommon promoter structure for Mlc1, with CCAAT-boxes representing the only noticeable elements.

  1. 缺氧复氧后内皮细胞中PBEF与VEGF、p-MLC的相关性研究%Study on the Relationship between p-MLC and VEGF and PBEF in Endothelial Cells after Hypoxia

    Institute of Scientific and Technical Information of China (English)

    杨威; 董啸

    2015-01-01

    目的探讨前β细胞克隆增强因子(PBEF)、血管内皮生长因子(VEGF)和肌球蛋白轻链磷酸化(pMLC)在人脐静脉内皮细胞(HUVEC)缺氧复氧中的表达及相互间的相关性。方法体外常规培养HUVEC细胞,实验分为对照组、缺氧组和缺氧复氧组三组,每组10个培养板孔细胞;Western blot ing检测三组PBEF、VEGF和pMLC蛋白表达;并分析PBEF、VEGF和pMLC三组在HUVEC缺氧复氧中的相关性。结果 HUVEC细胞缺氧后,PBEF,VEGF和pMLC蛋白表达较对照组均显著升高(<0.01);缺氧组HUVEC细胞复氧12h后,PBEF,VEGF和pMLC蛋白表达较单纯缺氧组进一步升高,比较差异有统计学意义(<0.01)。 PBEF、VEGF和pMLC蛋白在HUVEC缺氧复氧中的表达均呈正相关;PBEF和VEGF间的等级相关系数为0.493,PBEF和pMLC间的等级相关系数为0.534,VEGF和pMLC间的等级相关系数为0.469。结论 PBEF、VEGF和pMLC参与HUVEC缺氧复氧过程,且三者在HUVEC缺氧复氧中的表达之间均呈正相关。%Objective To discuss the expression and relationship of pre-βcel colony-enhancing factor(PBEF),vessel endothelial growth factor(VEGF)and phosphorylation of myosin light chains(pMLC)in anoxia-reoxygenation of human umbilical vein endothelial cel (HUVEC).Methods HUVEC cel s were cultivated by the conventional method in vitro and divided into control,anoxia and anoxia-reoxygenation groups with 10 hole cel culture plate each.Expression of PBEF,VEGF and pMLC protein in every group were detected by western blot ing and analyzed the cor elation in anoxia-reoxygenation of HUVEC.Results Expression of PBEF,VEGF and pMLC protein were obviously higher than control group after HUVEC cel s of oxygen ( <0.01).Expression of PBEF,VEGF and pMLC protein increased more in HUVEC cel s of anoxia group after reoxygenation 12 h than anoxia group with statistical y significances ( <0.01).There are positive cor elations among expression of PBEF, VEGF and pMLC protein in HUVEC anoxia

  2. Evaluation of dosimetric effect caused by slowing with multi-leaf collimator (MLC) leaves for volumetric modulated arc therapy (VMAT)

    OpenAIRE

    2016-01-01

    Background This study is to report 1) the sensitivity of intensity modulated radiation therapy (IMRT) QA method for clinical volumetric modulated arc therapy (VMAT) plans with multi-leaf collimator (MLC) leaf errors that will not trigger MLC interlock during beam delivery; 2) the effect of non-beam-hold MLC leaf errors on the quality of VMAT plan dose delivery. Materials and methods. Eleven VMAT plans were selected and modified using an in-house developed software. For each control point of a...

  3. Heterologous expression of MlcE in Saccharomyces cerevisiae provides resistance to natural and semi-synthetic statins

    DEFF Research Database (Denmark)

    Ley, Ana; Coumou, Hilde Cornelijne; Frandsen, Rasmus John Normand

    2015-01-01

    -encoding gene mlcE from the mevastatin-producing Penicillium citrinum into the S. cerevisiae genome. The resulting strain showed increased resistance to both natural statins (mevastatin and lovastatin) and semi-synthetic statin (simvastatin) when compared to the wild type strain. Expression of RFP-tagged mlc......E showed that MlcE is localized to the yeast plasma and vacuolar membranes. We provide a possible engineering strategy for improvement of future yeast based production of natural and semi-synthetic statins....

  4. SU-E-T-01: 2-D Characterization of DLG Among All MLC Leaf Pairs

    Energy Technology Data Exchange (ETDEWEB)

    Kumaraswamy, L; Xu, Z; Podgorsak, M [Roswell Park Cancer Institute, Buffalo, NY (United States); Schmitt, J [RadAmerica, LLC--MedStar Health, Baltimore, MD (United States); Bailey, D [Northside Hospital, Atlanta, GA (United States)

    2014-06-01

    Purpose: The aim of this study is to evaluate the variation of dosimetric leaf-gap (DLG) along the travel path of each MLC leaf pair. This study evaluates whether the spatial variations in DLG could cause dose differences between TPS-calculated and measured dose. Methods: The 6MV DLG values were measured for all leaf pairs in the direction of leaf motion using a 2-D diode array and 0.6cc ion chamber. These measurements were performed on two Varian Linacs, employing the Millennium 120-leaf MLC and a 2-D-DLG variation map was created via in-house software. Several test plans were created with sweeping MLC fields using constant gaps from 2mm to 10mm and corrected for 2-D variation utilizing in-house software. Measurements were performed utilizing the MapCHECK at 5.0cm depth for plans with and without the 2-D DLG correction and compared to the TPS calculated dose via gamma analysis (3%/3mm). Results: The measured DLGs for the middle 40 MLC leaf pairs (0.5cm width) were very similar along the central superior-inferior axis, with maximum variation of 0.2mm. The outer 20 MLC leaf pairs (1.0cm width) have DLG values from 0.32mm (mean) to 0.65mm (maximum) lower than the central leaf-pair, depending on off-axis distance. Gamma pass rates for the 2mm, 4mm, and 6mm sweep plans increased by 23.2%, 28.7%, and 26.0% respectively using the 2-D-DLG correction. The most improved dose points occur in areas modulated by the 1.0cm leaf-pairs. The gamma pass rate for the 10mm sweep plan increased by only 7.7%, indicating that the 2D variation becomes less significant for dynamic plans with larger MLC gaps. Conclusion: Fluences residing significantly off-axis with narrow sweeping gaps may exhibit significant variations from planned dose due to large differences between the true DLG exhibited by the 1.0cm leaf-pairs versus the constant DLG value utilized by the TPS for dose calculation.

  5. The role of AGFA high-energy CR in the calibration and quality control of multileaf collimators (MLC)

    Institute of Scientific and Technical Information of China (English)

    LI Zhaobin; XIONG Fei; HUANG Guofeng; CAO Zheng; JIANG Ruiyao; FU Shen

    2007-01-01

    Linear accelerators equipped with multileaf collimators (MLC) are becoming more common and are widely used in the intensity-modulated radiotherapy (IMRT). There is an imperative need to ensure the commissioning specification of the linear accelerators for the sake of quality assurance (QA) and quality control (QC). This paper is aimed to investigate the role of AGFA high-energy CR (Computed Radiography) in calibrating dynamic multileaf collimators and evaluating the accuracy of the leaf position. The result shows that AGFA high-energy CR can easily and conveniently be used to calibrate MLC and verify its position. Hence, the application of AGFA high-energy CR is proved to be an accurate and time-saving method for routine MLC QC, especially when MLC calibration adjustments are required.

  6. The dosimetric impact of inversely optimized arc radiotherapy plan modulation for real-time dynamic MLC tracking delivery

    DEFF Research Database (Denmark)

    Falk, Marianne; Larsson, Tobias; Keall, Paul

    2012-01-01

    -to-peak displacement of 2 cm and a cycle time of 6 s. The delivery was adjusted to the target motion using MLC tracking, guided in real-time by an infrared optical system. The dosimetric results were evaluated using gamma index evaluation with static target measurements as reference. Results: The plan quality......Purpose: Real-time dynamic multileaf collimator (MLC) tracking for management of intrafraction tumor motion can be challenging for highly modulated beams, as the leaves need to travel far to adjust for target motion perpendicular to the leaf travel direction. The plan modulation can be reduced...... on the dosimetric accuracy of MLC tracking delivery. Specifically, the possibility of predicting the accuracy of MLC tracking delivery based on the plan modulation was investigated. Methods: Inversely optimized arc radiotherapy plans were created on CT-data of three lung cancer patients. For each case, five plans...

  7. Motion management during IMAT treatment of mobile lung tumors-A comparison of MLC tracking and gated delivery

    DEFF Research Database (Denmark)

    Falk, Marianne; Pommer, Tobias; Keall, Paul

    2014-01-01

    Purpose:To compare real-time dynamic multileaf collimator (MLC) tracking, respiratory amplitude and phase gating, and no compensation for intrafraction motion management during intensity modulated arc therapy (IMAT). Methods: Motion management with MLC tracking and gating was evaluated for four l...... longer delivery time. In a clinical setting, the optical monitoring of the patients breathing would have to be correlated to the internal movements of the tumor...

  8. Real-time auto-adaptive margin generation for MLC-tracked radiotherapy

    Science.gov (United States)

    Glitzner, M.; Fast, M. F.; de Senneville, B. Denis; Nill, S.; Oelfke, U.; Lagendijk, J. J. W.; Raaymakers, B. W.; Crijns, S. P. M.

    2017-01-01

    In radiotherapy, abdominal and thoracic sites are candidates for performing motion tracking. With real-time control it is possible to adjust the multileaf collimator (MLC) position to the target position. However, positions are not perfectly matched and position errors arise from system delays and complicated response of the electromechanic MLC system. Although, it is possible to compensate parts of these errors by using predictors, residual errors remain and need to be compensated to retain target coverage. This work presents a method to statistically describe tracking errors and to automatically derive a patient-specific, per-segment margin to compensate the arising underdosage on-line, i.e. during plan delivery. The statistics of the geometric error between intended and actual machine position are derived using kernel density estimators. Subsequently a margin is calculated on-line according to a selected coverage parameter, which determines the amount of accepted underdosage. The margin is then applied onto the actual segment to accommodate the positioning errors in the enlarged segment. The proof-of-concept was tested in an on-line tracking experiment and showed the ability to recover underdosages for two test cases, increasing {{V}90 %} in the underdosed area about 47 % and 41 % , respectively. The used dose model was able to predict the loss of dose due to tracking errors and could be used to infer the necessary margins. The implementation had a running time of 23 ms which is compatible with real-time requirements of MLC tracking systems. The auto-adaptivity to machine and patient characteristics makes the technique a generic yet intuitive candidate to avoid underdosages due to MLC tracking errors.

  9. Inhibition of MLC phosphorylation restricts replication of influenza virus--a mechanism of action for anti-influenza agents.

    Directory of Open Access Journals (Sweden)

    Mehran Haidari

    Full Text Available Influenza A viruses are a severe threat worldwide, causing large epidemics that kill thousands every year. Prevention of influenza infection is complicated by continuous viral antigenic changes. Newer anti-influenza agents include MEK/ERK and protein kinase C inhibitors; however, the downstream effectors of these pathways have not been determined. In this study, we identified a common mechanism for the inhibitory effects of a significant group of anti-influenza agents. Our studies showed that influenza infection activates a series of signaling pathways that converge to induce myosin light chain (MLC phosphorylation and remodeling of the actin cytoskeleton. Inhibiting MLC phosphorylation by blocking RhoA/Rho kinase, phospholipase C/protein kinase C, and HRas/Raf/MEK/ERK pathways with the use of genetic or chemical manipulation leads to the inhibition of influenza proliferation. In contrast, the induction of MLC phosphorylation enhances influenza proliferation, as does activation of the HRas/Raf/MEK/ERK signaling pathway. This effect is attenuated by inhibiting MLC phosphorylation. Additionally, in intracellular trafficking studies, we found that the nuclear export of influenza ribonucleoprotein depends on MLC phosphorylation. Our studies provide evidence that modulation of MLC phosphorylation is an underlying mechanism for the inhibitory effects of many anti-influenza compounds.

  10. The first patient treatment of electromagnetic-guided real time adaptive radiotherapy using MLC tracking for lung SABR.

    Science.gov (United States)

    Booth, Jeremy T; Caillet, Vincent; Hardcastle, Nicholas; O'Brien, Ricky; Szymura, Kathryn; Crasta, Charlene; Harris, Benjamin; Haddad, Carol; Eade, Thomas; Keall, Paul J

    2016-10-01

    Real time adaptive radiotherapy that enables smaller irradiated volumes may reduce pulmonary toxicity. We report on the first patient treatment of electromagnetic-guided real time adaptive radiotherapy delivered with MLC tracking for lung stereotactic ablative body radiotherapy. A clinical trial was developed to investigate the safety and feasibility of MLC tracking in lung. The first patient was an 80-year old man with a single left lower lobe lung metastasis to be treated with SABR to 48Gy in 4 fractions. In-house software was integrated with a standard linear accelerator to adapt the treatment beam shape and position based on electromagnetic transponders implanted in the lung. MLC tracking plans were compared against standard ITV-based treatment planning. MLC tracking plan delivery was reconstructed in the patient to confirm safe delivery. Real time adaptive radiotherapy delivered with MLC tracking compared to standard ITV-based planning reduced the PTV by 41% (18.7-11cm(3)) and the mean lung dose by 30% (202-140cGy), V20 by 35% (2.6-1.5%) and V5 by 9% (8.9-8%). An emerging technology, MLC tracking, has been translated into the clinic and used to treat lung SABR patients for the first time. This milestone represents an important first step for clinical real-time adaptive radiotherapy that could reduce pulmonary toxicity in lung radiotherapy. Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.

  11. Fast leaf-fitting with generalized underdose/overdose constraints for real-time MLC tracking

    Science.gov (United States)

    Ruan, Dan; Sawant, Amit

    2016-01-01

    Purpose: Real-time multileaf collimator (MLC) tracking is a promising approach to the management of intrafractional tumor motion during thoracic and abdominal radiotherapy. MLC tracking is typically performed in two steps: transforming a planned MLC aperture in response to patient motion and refitting the leaves to the newly generated aperture. One of the challenges of this approach is the inability to faithfully reproduce the desired motion-adapted aperture. This work presents an optimization-based framework with which to solve this leaf-fitting problem in real-time. Methods: This optimization framework is designed to facilitate the determination of leaf positions in real-time while accounting for the trade-off between coverage of the PTV and avoidance of organs at risk (OARs). Derived within this framework, an algorithm is presented that can account for general linear transformations of the planned MLC aperture, particularly 3D translations and in-plane rotations. This algorithm, together with algorithms presented in Sawant et al. [“Management of three-dimensional intrafraction motion through real-time DMLC tracking,” Med. Phys. 35, 2050–2061 (2008)] and Ruan and Keall [Presented at the 2011 IEEE Power Engineering and Automation Conference (PEAM) (2011) (unpublished)], was applied to apertures derived from eight lung intensity modulated radiotherapy plans subjected to six-degree-of-freedom motion traces acquired from lung cancer patients using the kilovoltage intrafraction monitoring system developed at the University of Sydney. A quality-of-fit metric was defined, and each algorithm was evaluated in terms of quality-of-fit and computation time. Results: This algorithm is shown to perform leaf-fittings of apertures, each with 80 leaf pairs, in 0.226 ms on average as compared to 0.082 and 64.2 ms for the algorithms of Sawant et al., Ruan, and Keall, respectively. The algorithm shows approximately 12% improvement in quality-of-fit over the Sawant et al

  12. Real-time tumor tracking: Automatic compensation of target motion using the Siemens 160 MLC

    Energy Technology Data Exchange (ETDEWEB)

    Tacke, Martin B.; Nill, Simeon; Krauss, Andreas; Oelfke, Uwe [Department of Medical Physics in Radiation Oncology, German Cancer Research Center, Im Neuenheimer Feld 280, D-69120 Heidelberg (Germany)

    2010-02-15

    Purpose: Advanced high quality radiation therapy techniques such as IMRT require an accurate delivery of precisely modulated radiation fields to the target volume. Interfractional and intrafractional motion of the patient's anatomy, however, may considerably deteriorate the accuracy of the delivered dose to the planned dose distributions. In order to compensate for these potential errors, a dynamic real-time capable MLC control system was designed. Methods: The newly developed adaptive MLC control system contains specialized algorithms which are capable of continuous optimization and correction of the aperture of the MLC according to the motion of the target volume during the dose delivery. The algorithms calculate the new leaf positions based on target information provided online to the system. The algorithms were implemented in a dynamic target tracking control system designed for a Siemens 160 MLC. To assess the quality of the new target tracking system in terms of dosimetric accuracy, experiments with various types of motion patterns using different phantom setups were performed. The phantoms were equipped with radiochromic films placed between solid water slabs. Dosimetric results of exemplary deliveries to moving targets with and without dynamic MLC tracking applied were compared in terms of the gamma criterion to the reference dose delivered to a static phantom. Results: Our measurements indicated that dose errors for clinically relevant two-dimensional target motion can be compensated by the new control system during the dose delivery of open fields. For a clinical IMRT dose distribution, the gamma success rate was increased from 19% to 77% using the new tracking system. Similar improvements were achieved for the delivery of a complete IMRT treatment fraction to a moving lung phantom. However, dosimetric accuracy was limited by the system's latency of 400 ms and the finite leaf width of 5 mm in the isocenter plane. Conclusions: Different

  13. Treatment plan comparison between helical tomotherapy and MLC-based IMRT using radiobiological measures

    Energy Technology Data Exchange (ETDEWEB)

    Mavroidis, Panayiotis [Department of Medical Radiation Physics, Karolinska Institutet and Stockholm University (Sweden); Ferreira, Brigida Costa [Department of Medical Radiation Physics, Karolinska Institutet and Stockholm University (Sweden); Shi, Chengyu [Department of Radiation Oncology, University of Texas Health Science Center, San Antonio, TX (United States); Lind, Bengt K [Department of Medical Radiation Physics, Karolinska Institutet and Stockholm University (Sweden); Papanikolaou, Nikos [Department of Radiation Oncology, University of Texas Health Science Center, San Antonio, TX (United States)

    2007-07-07

    The rapid implementation of advanced treatment planning and delivery technologies for radiation therapy has brought new challenges in evaluating the most effective treatment modality. Intensity-modulated radiotherapy (IMRT) using multi-leaf collimators (MLC) and helical tomotherapy (HT) are becoming popular modes of treatment delivery and their application and effectiveness continues to be investigated. Presently, there are several treatment planning systems (TPS) that can generate and optimize IMRT plans based on user-defined objective functions for the internal target volume (ITV) and organs at risk (OAR). However, the radiobiological parameters of the different tumours and normal tissues are typically not taken into account during dose prescription and optimization of a treatment plan or during plan evaluation. The suitability of a treatment plan is typically decided based on dosimetric criteria such as dose-volume histograms (DVH), maximum, minimum, mean and standard deviation of the dose distribution. For a more comprehensive treatment plan evaluation, the biologically effective uniform dose D-bar is applied together with the complication-free tumour control probability (P{sub +}). Its utilization is demonstrated using three clinical cases that were planned with two different forms of IMRT. In this study, three different cancer types at different anatomical sites were investigated: head and neck, lung and prostate cancers. For each cancer type, a linac MLC-based step-and-shoot IMRT plan and a HT plan were developed. The MLC-based IMRT treatment plans were developed on the Philips treatment-planning platform, using the Pinnacle 7.6 software release. For the tomotherapy HiArt plans, the dedicated tomotherapy treatment planning station was used, running version 2.1.2. By using D-bar as the common prescription point of the treatment plans and plotting the tissue response probabilities versus D-bar for a range of prescription doses, a number of plan trials can be

  14. SU-E-T-506: Dosimetric Verification of Photon MLC Delivered Electron Fields for Implementing MERT On An Artiste Linac

    Energy Technology Data Exchange (ETDEWEB)

    Jin, L; Eldib, A; Li, J; Wang, L; Ma, C [Fox Chase Cancer Center, Philadelphia, PA (United States); Fan, J [Virtua Fox Chase Cancer Center, Philadelphia, PA (United States)

    2014-06-01

    Purpose: To verify the dose accuracy of photon MLC delivered electron fields for implementing energy-intensity modulated electron radiotherapy (MERT) on an Artiste linac. Methods: It was proposed to deliver MERT on an Artiste linac at a short SSD (60 cm) to reduce beam penumbra caused by electron scatters. An in-house developed Monte Carlo (MC)-based dose calculation/optimization planning code was used for treatment planning. Our previous study showed that the measured dose distribution of a breast plan showed good agreement with the calculations in low-medium dose regions while the differences in high dose regions were outstanding. A continuous work found that the discrepancy is mainly caused by improper modeling in MC for the single focused MLC in the Artiste which was simplified as double focused in the previous MC simulations. With this remodeled MLC in the calculations, an energy-intensity modulated electron plan using 6, 9, 12 and 15 MeV was generated for a breast treatment on a breast phantom at a 60 cm SSD and recalculated regarding a solid water phantom. For a test study, four of MLC segments (each with a different energy) generated in the plan were delivered to the phantom and a film measurement was performed at the depth of 2 cm. The measured 2D dose distribution was then compared with calculations. Results: For composite doses of the four segments, measured 2D dose distributions overall agree well with the calculations (3mm/3%) in most area. The separate measurement for a single MLC segment for each of energies also showed the consistence with the calculations. Conclusion: After remodeling MLC in the MC calculations, the measured dose distribution for a subset of MLC segments from a MERT plan showed good agreement. Further detailed verification for the full plan will be the work in the next step.

  15. T-cell receptor v-alpha and v-Beta gene usage in interleukin-2-cultured tumor-infiltrating lymphocytes from patients with breast-cancer

    DEFF Research Database (Denmark)

    Andersen, E; Scholler, J; Straten, P;

    1994-01-01

    surface through the T cell receptor (TCR) complex. We have studied the phenotype, cytotoxicity, and expression of TCR variable (V) alpha and beta chain on in vitro IL-2-cultured TIL isolated from primary malignant breast tumors from 11 patients. 10/11 cultures were dominated by CD4(+) (T-helper) cells....... The different TIL cultures exhibited varying levels of cytotoxicity against the natural killer (NK)-sensitive cell line K562 and breast cancer cell line T47D. The level of clonality, as measured by PCR-based analyses of usage of the different V segments was low, as only a few tumors showed patterns...... of restricted V gene expression. The mean number of V alpha segments per TIL culture was higher than the number of V beta segments per culture. A significant negative correlation was observed between the number of CD4+ cells and the number of V beta segments per culture, and no other correlations between...

  16. Determination of TiO2, ZrO2, and Al2O3 nanoparticles on genotoxic responses in human peripheral blood lymphocytes and cultured embyronic kidney cells.

    Science.gov (United States)

    Demir, Eşref; Burgucu, Durmuş; Turna, Fatma; Aksakal, Sezgin; Kaya, Bülent

    2013-01-01

    In this study a genotoxic evaluation of titanium dioxide (TiO2, 2.3 nm), zirconium oxide (ZrO2, 6 nm), aluminum oxide (Al2O3, 16.7 nm) nanoparticles (NP) and their ionic forms was conducted using human peripheral blood lymphocytes and cultured human embryonic kidney (HEK293) cells by means of a modified alkaline comet assay with/without the formamidopyrimidine-DNA glycosylase (Fpg) and endonuclease III (Endo III) enzymes. Modifications to the comet assay by using lesion-specific endonucleases, such as Endo III and Fpg, detect DNA bases with oxidative damage. Both human peripheral blood lymphocytes and cultured embryonic kidney cells were incubated with TiO2, ZrO2, or Al2O3 NP at concentrations of 1, 10, or 100 μg/ml. Our results showed no significant induction in DNA damage by the comet assay with/without the Endo III and Fpg enzymes at all concentrations of ZrO2 and Al2O3. In the case of TiO2 NP only the highest concentration of 100 μg/ml significantly induced a genotoxic response. Data thus indicate that both ZrO2 and Al2O3 NP were not genotoxic in our system and in the case of TiO2 the lowest-observed-adverse-effect level (LOAEL) for genotoxicity was 100 μg/ml. Evidence indicates that these metallic NP are considered safe in light of the fact that no genotoxicity was noted with ZrO2 and Al2O3 and that the highest TiO2 concentration is not environmentally relevant.

  17. Development and evaluation of aperture-based complexity metrics using film and EPID measurements of static MLC openings

    Energy Technology Data Exchange (ETDEWEB)

    Götstedt, Julia [Department of Radiation Physics, University of Gothenburg, Göteborg 413 45 (Sweden); Karlsson Hauer, Anna; Bäck, Anna, E-mail: anna.back@vgregion.se [Department of Therapeutic Radiation Physics, Sahlgrenska University Hospital, Göteborg 413 45 (Sweden)

    2015-07-15

    Purpose: Complexity metrics have been suggested as a complement to measurement-based quality assurance for intensity modulated radiation therapy (IMRT) and volumetric modulated arc therapy (VMAT). However, these metrics have not yet been sufficiently validated. This study develops and evaluates new aperture-based complexity metrics in the context of static multileaf collimator (MLC) openings and compares them to previously published metrics. Methods: This study develops the converted aperture metric and the edge area metric. The converted aperture metric is based on small and irregular parts within the MLC opening that are quantified as measured distances between MLC leaves. The edge area metric is based on the relative size of the region around the edges defined by the MLC. Another metric suggested in this study is the circumference/area ratio. Earlier defined aperture-based complexity metrics—the modulation complexity score, the edge metric, the ratio monitor units (MU)/Gy, the aperture area, and the aperture irregularity—are compared to the newly proposed metrics. A set of small and irregular static MLC openings are created which simulate individual IMRT/VMAT control points of various complexities. These are measured with both an amorphous silicon electronic portal imaging device and EBT3 film. The differences between calculated and measured dose distributions are evaluated using a pixel-by-pixel comparison with two global dose difference criteria of 3% and 5%. The extent of the dose differences, expressed in terms of pass rate, is used as a measure of the complexity of the MLC openings and used for the evaluation of the metrics compared in this study. The different complexity scores are calculated for each created static MLC opening. The correlation between the calculated complexity scores and the extent of the dose differences (pass rate) are analyzed in scatter plots and using Pearson’s r-values. Results: The complexity scores calculated by the edge

  18. Ofatumumab, Pentostatin, and Cyclophosphamide in Treating Patients With Untreated Chronic Lymphocytic Leukemia or Small Lymphocytic Lymphoma

    Science.gov (United States)

    2014-10-30

    Hematopoietic/Lymphoid Cancer; B-cell Chronic Lymphocytic Leukemia; Contiguous Stage II Small Lymphocytic Lymphoma; Noncontiguous Stage II Small Lymphocytic Lymphoma; Stage 0 Chronic Lymphocytic Leukemia; Stage I Chronic Lymphocytic Leukemia; Stage I Small Lymphocytic Lymphoma; Stage II Chronic Lymphocytic Leukemia; Stage III Chronic Lymphocytic Leukemia; Stage III Small Lymphocytic Lymphoma; Stage IV Chronic Lymphocytic Leukemia; Stage IV Small Lymphocytic Lymphoma

  19. SU-E-J-67: Evaluation of Adaptive MLC Morphing for Online Correction of Prostate Cancer Radiotherapy

    Energy Technology Data Exchange (ETDEWEB)

    Sandhu, R; Qin, A; Yan, D [William Beaumont Hospital, Royal Oak, MI (United States)

    2015-06-15

    Purpose: Online adaptive MLC morphing is desirable over translational couch shifts to accommodate target position as well as anatomic changes. A reliable method of adaptive MLC segment to target during prostate cancer IMRT treatment is proposed and evaluated by comparison with daily online-image guidance (IGRT) correction and online-IMRT planning. Methods: The MLC adaptive algorithm involves following steps; move the MLC segments according to target translational shifts, and then morph the segment shape to maintain the spatial relationship between the planning-target contour and MLC segment. Efficacy of this method was evaluated retrospectively using daily-CBCT images on seven prostate patients treated with seven-beam IMRT treatment to deliver 64Gy in 20 fractions. Daily modification was simulated with three approaches; daily-IGRT correction based on implanted radio-markers, adaptive MLC morphing, and online-IMRT planning, with no-residual variation. The selected dosimetric endpoints and nEUD (normalized equivalent uniform dose to online-IMRT planning) of each organ of interest were determined for evaluation and comparison. Results: For target(prostate), bladder and rectal-wall, the mean±sd of nEUD were 97.6%+3.2%, 103.9%±4.9% and 97.4%±1.1% for daily-IGRT correction; and 100.2%+0.2%, 108.9%±5.1% and 99.8%±1.2% for adaptive MLC morphing, respectively. For daily-IGRT correction, adaptive MLC morphing and online-IMRT planning, target D99 was <95% of the prescription dose in 30%, 0% and 0% of 140 fractions, respectively. For the rectal-wall, D5 exceeded 105% of the planned-D5 in 2.8%, 11.4% and 0% of 140 fractions, respectively. For the bladder, Dmax exceeded 105% of the planned-D5 in 2.8%, 5.6% and 0% of 140 fractions, respectively. D30 of bladder and rectal-wall were well within the planned-D30 for all three approaches. Conclusion: The proposed method of adaptive MLC morphing can be beneficial for the prostate patient population with large deformation and

  20. Monitoring daily MLC positional errors using trajectory log files and EPID measurements for IMRT and VMAT deliveries

    Science.gov (United States)

    Agnew, A.; Agnew, C. E.; Grattan, M. W. D.; Hounsell, A. R.; McGarry, C. K.

    2014-05-01

    This work investigated the differences between multileaf collimator (MLC) positioning accuracy determined using either log files or electronic portal imaging devices (EPID) and then assessed the possibility of reducing patient specific quality control (QC) via phantom-less methodologies. In-house software was developed, and validated, to track MLC positional accuracy with the rotational and static gantry picket fence tests using an integrated electronic portal image. This software was used to monitor MLC daily performance over a 1 year period for two Varian TrueBeam linear accelerators, with the results directly compared with MLC positions determined using leaf trajectory log files. This software was validated by introducing known shifts and collimator errors. Skewness of the MLCs was found to be 0.03 ± 0.06° (mean ±1 standard deviation (SD)) and was dependent on whether the collimator was rotated manually or automatically. Trajectory log files, analysed using in-house software, showed average MLC positioning errors with a magnitude of 0.004 ± 0.003 mm (rotational) and 0.004 ± 0.011 mm (static) across two TrueBeam units over 1 year (mean ±1 SD). These ranges, as indicated by the SD, were lower than the related average MLC positioning errors of 0.000 ± 0.025 mm (rotational) and 0.000 ± 0.039 mm (static) that were obtained using the in-house EPID based software. The range of EPID measured MLC positional errors was larger due to the inherent uncertainties of the procedure. Over the duration of the study, multiple MLC positional errors were detected using the EPID based software but these same errors were not detected using the trajectory log files. This work shows the importance of increasing linac specific QC when phantom-less methodologies, such as the use of log files, are used to reduce patient specific QC. Tolerances of 0.25 mm have been created for the MLC positional errors using the EPID-based automated picket fence test. The software allows diagnosis

  1. SU-E-T-583: Optimizing the MLC Model Parameters for IMRT in the RayStation Treatment Planning System

    Energy Technology Data Exchange (ETDEWEB)

    Chen, S; Yi, B; Xu, H; Yang, X; Prado, K; D' Souza, W [University of Maryland School of Medicine, Baltimore, MD (United States)

    2014-06-01

    Purpose: To optimize the MLC model parameters for IMRT in the RayStation v.4.0 planning system and for a Varian C-series Linac with a 120-leaf Millennium MLC. Methods: The RayStation treatment planning system models rounded leaf-end MLC with the following parameters: average transmission, leaf-tip width, tongue-and-groove, and position offset. The position offset was provided by Varian. The leaf-tip width was iteratively evaluated by comparing computed and measured transverse dose profiles of MLC-defined fields at dmax in water. The profile comparison was also used to verify the MLC position offset. The transmission factor and leaf tongue width were derived iteratively by optimizing five clinical patient IMRT QA Results: brain, lung, pancreas, head-and-neck (HN), and prostate. The HN and prostate cases involved splitting fields. Verifications were performed with Mapcheck2 measurements and Monte Carlo calculations. Finally, the MLC model was validated using five test IMRT cases from the AAPM TG119 report. Absolute gamma analyses (3mm/3% and 2mm/2%) were applied. In addition, computed output factors for MLC-defined small fields (2×2, 3×3, 4×4, 6×6cm) of both 6MV and 18MV were compared to those measured by the Radiological Physics Center (RPC). Results: Both 6MV and 18MV models were determined to have the same MLC parameters: 2.5% transmission, tongue-and-groove 0.05cm, and leaftip 0.3cm. IMRT QA analysis for five cases in TG119 resulted in a 100% passing rate with 3mm/3% gamma analysis for 6MV, and >97.5% for 18MV. With 2mm/2% gamma analysis, the passing rate was >94.6% for 6MV and >90.9% for 18MV. The difference between computed output factors in RayStation and RPC measurements was less than 2% for all MLCdefined fields, which meets the RPC's acceptance criterion. Conclusion: The rounded leaf-end MLC model in RayStation 4.0 planning system was verified and IMRT commissioning was clinically acceptable. The IMRT commissioning was well validated using guidance

  2. Determination of the optimal tolerance for MLC positioning in sliding window and VMAT techniques

    Energy Technology Data Exchange (ETDEWEB)

    Hernandez, V., E-mail: vhernandezmasgrau@gmail.com; Abella, R. [Department of Medical Physics, Hospital Sant Joan de Reus, IISPV, Tarragona 43204 (Spain); Calvo, J. F. [Department of Radiation Oncology, Hospital Quirón, Barcelona 08023 (Spain); Jurado-Bruggemann, D. [Department of Medical Physics, Institut Català d’Oncologia, Girona 17007 (Spain); Sancho, I. [Department of Medical Physics, Institut Català d’Oncologia, L’Hospitalet de Llobregat 08908 (Spain); Carrasco, P. [Department of Medical Physics, Hospital de la Santa Creu i Sant Pau, Barcelona 08041 (Spain)

    2015-04-15

    Purpose: Several authors have recommended a 2 mm tolerance for multileaf collimator (MLC) positioning in sliding window treatments. In volumetric modulated arc therapy (VMAT) treatments, however, the optimal tolerance for MLC positioning remains unknown. In this paper, the authors present the results of a multicenter study to determine the optimal tolerance for both techniques. Methods: The procedure used is based on dynalog file analysis. The study was carried out using seven Varian linear accelerators from five different centers. Dynalogs were collected from over 100 000 clinical treatments and in-house software was used to compute the number of tolerance faults as a function of the user-defined tolerance. Thus, the optimal value for this tolerance, defined as the lowest achievable value, was investigated. Results: Dynalog files accurately predict the number of tolerance faults as a function of the tolerance value, especially for low fault incidences. All MLCs behaved similarly and the Millennium120 and the HD120 models yielded comparable results. In sliding window techniques, the number of beams with an incidence of hold-offs >1% rapidly decreases for a tolerance of 1.5 mm. In VMAT techniques, the number of tolerance faults sharply drops for tolerances around 2 mm. For a tolerance of 2.5 mm, less than 0.1% of the VMAT arcs presented tolerance faults. Conclusions: Dynalog analysis provides a feasible method for investigating the optimal tolerance for MLC positioning in dynamic fields. In sliding window treatments, the tolerance of 2 mm was found to be adequate, although it can be reduced to 1.5 mm. In VMAT treatments, the typically used 5 mm tolerance is excessively high. Instead, a tolerance of 2.5 mm is recommended.

  3. Improving IMRT-plan quality with MLC leaf position refinement post plan optimization.

    Science.gov (United States)

    Niu, Ying; Zhang, Guowei; Berman, Barry L; Parke, William C; Yi, Byongyong; Yu, Cedric X

    2012-08-01

    In intensity-modulated radiation therapy (IMRT) planning, reducing the pencil-beam size may lead to a significant improvement in dose conformity, but also increase the time needed for the dose calculation and plan optimization. The authors develop and evaluate a postoptimization refinement (POpR) method, which makes fine adjustments to the multileaf collimator (MLC) leaf positions after plan optimization, enhancing the spatial precision and improving the plan quality without a significant impact on the computational burden. The authors' POpR method is implemented using a commercial treatment planning system based on direct aperture optimization. After an IMRT plan is optimized using pencil beams with regular pencil-beam step size, a greedy search is conducted by looping through all of the involved MLC leaves to see if moving the MLC leaf in or out by half of a pencil-beam step size will improve the objective function value. The half-sized pencil beams, which are used for updating dose distribution in the greedy search, are derived from the existing full-sized pencil beams without need for further pencil-beam dose calculations. A benchmark phantom case and a head-and-neck (HN) case are studied for testing the authors' POpR method. Using a benchmark phantom and a HN case, the authors have verified that their POpR method can be an efficient technique in the IMRT planning process. Effectiveness of POpR is confirmed by noting significant improvements in objective function values. Dosimetric benefits of POpR are comparable to those of using a finer pencil-beam size from the optimization start, but with far less computation and time. The POpR is a feasible and practical method to significantly improve IMRT-plan quality without compromising the planning efficiency.

  4. Specific proliferative response of human lymphocytes to purified soluble antigens from Plasmodium falciparum in vitro cultures and to antigens from malaria patients' sera

    DEFF Research Database (Denmark)

    Bygbjerg, I C; Jepsen, S; Theander, T G;

    1985-01-01

    . The responses were strongest with lymphocytes from individuals infected with falciparum and ovale malaria; vivax malaria infections induced a lower level of response and lymphocytes of unsensitized individuals were little affected. Lymphocytes from unsensitized individuals did not respond to the affinity...

  5. 金属硫蛋白对热应激下体外培养奶牛淋巴细胞的影响%Effect of metallothionein on lymphocyte apoptosis of the dairy cow cultured in vitro under heat stress

    Institute of Scientific and Technical Information of China (English)

    占今舜; 吴力专; 李丽立; 张彬

    2014-01-01

    本研究的目的在于探讨外源性金属硫蛋白(MT)对热应激下体外培养奶牛淋巴细胞凋亡的影响。将屠宰获取的奶牛脾脏淋巴细胞随机分为4组,每组细胞培养液中MT的终浓度分别为0μg/mL(对照组)、70μg/mL(第Ⅰ组)、140μg/mL(第Ⅱ组)、210μg/mL(第Ⅲ组),同时置于细胞培养箱(37℃、5%CO2)中培养20 h,之后同时置于43℃水浴锅中进行热应激处理1 h,再回细胞培养箱中培养3 h,然后利用形态学观察、RT-PCR等方法进行各项指标的检测。结果如下,1)通过形态学观察和AO/EB染色观察,表明添加MT可以有效缓解热应激对奶牛淋巴细胞的损伤,但缓解效果随 MT浓度的增加而下降;2)第Ⅰ~Ⅲ组的CAT活性、SOD活性和 GSH-Px活性均高于对照组(P<0.01),MDA含量均低于对照组(P<0.01),表明 MT能够有效改善淋巴细胞的抗氧化能力;3)试验Ⅰ~Ⅲ组的Bax基因相对表达量均显著低于对照组(P<0.01),但试验Ⅰ~Ⅲ组Bcl-2基因的表达量和p50基因的表达量均显著低于对照组(P<0.01),表明MT可以调节与细胞凋亡相关基因的表达。综上所述,添加MT可以降低奶牛淋巴细胞死亡率,改善奶牛淋巴细胞抗氧化性能,抑制奶牛淋巴细胞凋亡,以添加终浓度为70μg/mL的 MT效果最好。%The effect of exogenous MT on lymphocyte apoptosis of the dairy cow was studied invitro under heat stress.The splenic dairy cow lymphocytes were divided into 4 groups which had final concentrations of MT 0 (control group),70 (groupⅠ),140 (groupⅡ)and 210μg/mL (groupⅢ).The cells of each group were cul-tured in a cell incubator (37℃,5%CO2 ).After 20 h,the cells of each group were put into water bath pot (43℃)for 1 h,and then recultured in a cell incubator (37℃,5%CO2)for 3 h.Finally,various indexes were tested by observation of morphology and RT-PCR.1)MT added in cell

  6. Dosimetric comparison between cone/Iris-based and InCise MLC-based CyberKnife plans for single and multiple brain metastases.

    Science.gov (United States)

    Jang, Si Young; Lalonde, Ron; Ozhasoglu, Cihat; Burton, Steven; Heron, Dwight; Huq, M Saiful

    2016-09-08

    We performed an evaluation of the CyberKnife InCise MLC by comparing plan qualities for single and multiple brain lesions generated using the first version of InCise MLC, fixed cone, and Iris collimators. We also investigated differences in delivery efficiency among the three collimators. Twenty-four patients with single or multiple brain mets treated previously in our clinic on a CyberKnife M6 using cone/Iris collimators were selected for this study. Treatment plans were generated for all lesions using the InCise MLC. Number of monitor units, delivery time, target coverage, conformity index, and dose falloff were compared between MLC- and clinical cone/Iris-based plans. Statistical analysis was performed using the non-parametric Wilcoxon-Mann-Whitney signed-rank test. The planning accuracy of the MLC-based plans was validated using chamber and film measurements. The InCise MLC-based plans achieved mean dose and target coverage comparable to the cone/Iris-based plans. Although the conformity indices of the MLC-based plans were slightly higher than those of the cone/Iris-based plans, beam delivery time for the MLC-based plans was shorter by 30% ~ 40%. For smaller targets or cases with OARs located close to or abutting target volumes, MLC-based plans provided inferior dose conformity compared to cone/Iris-based plans. The QA results of MLC-based plans were within 5% absolute dose difference with over 90% gamma passing rate using 2%/2 mm gamma criteria. The first version of InCise MLC could be a useful delivery modality, especially for clinical situations for which delivery time is a limiting factor or for multitarget cases. © 2016 The Authors.

  7. Evaluation of calculation methods of collimator scatter factors in a linear accelerator equipped with MLC instead of lower collimators.

    Science.gov (United States)

    Kojima, Tomo; Sasaki, Koji

    2009-09-20

    In the monitor unit verification for high-energy radiation therapy, we evaluated methods of calculation of collimator scatter factors (S(c)) in a linear accelerator equipped with MLC instead of lower collimators. Routinely,S(c) is calculated from rectangular fields shaped by upper and lower jaws in the linear accelerator. However, this calculation method should not be used for the linear accelerator equipped with MLC instead of lower collimators. Consequently, we used a backprojected field at the flattening filter plane projected by calculation point's eye view on each MLC. We then attempted to deviseS(c) by using Clarkson's integration for these backprojected irregular fields. This method makes it possible to calculate collimator scatter factors in error of less than +/-0.3% in all of sixteen measured irregular fields.

  8. Motion management during IMAT treatment of mobile lung tumors—A comparison of MLC tracking and gated delivery

    Science.gov (United States)

    Falk, Marianne; Pommer, Tobias; Keall, Paul; Korreman, Stine; Persson, Gitte; Poulsen, Per; Munck af Rosenschöld, Per

    2014-01-01

    Purpose: To compare real-time dynamic multileaf collimator (MLC) tracking, respiratory amplitude and phase gating, and no compensation for intrafraction motion management during intensity modulated arc therapy (IMAT). Methods: Motion management with MLC tracking and gating was evaluated for four lung cancer patients. The IMAT plans were delivered to a dosimetric phantom mounted onto a 3D motion phantom performing patient-specific lung tumor motion. The MLC tracking system was guided by an optical system that used stereoscopic infrared (IR) cameras and five spherical reflecting markers attached to the dosimetric phantom. The gated delivery used a duty cycle of 35% and collected position data using an IR camera and two reflecting markers attached to a marker block. Results: The average gamma index failure rate (2% and 2 mm criteria) was delivery time. In a clinical setting, the optical monitoring of the patients breathing would have to be correlated to the internal movements of the tumor. PMID:25281946

  9. Bendamustine Plus Alemtuzumab for Refractory Chronic Lymphocytic Leukemia (CLL)

    Science.gov (United States)

    2013-08-20

    Recurrent Small Lymphocytic Lymphoma; Refractory Chronic Lymphocytic Leukemia; Stage III Chronic Lymphocytic Leukemia; Stage III Small Lymphocytic Lymphoma; Stage IV Chronic Lymphocytic Leukemia; Stage IV Small Lymphocytic Lymphoma

  10. Development of a Monte Carlo model for the Brainlab microMLC.

    Science.gov (United States)

    Belec, Jason; Patrocinio, Horacio; Verhaegen, Frank

    2005-03-07

    Stereotactic radiosurgery with several static conformal beams shaped by a micro multileaf collimator (microMLC) is used to treat small irregularly shaped brain lesions. Our goal is to perform Monte Carlo calculations of dose distributions for certain treatment plans as a verification tool. A dedicated microMLC component module for the BEAMnrc code was developed as part of this project and was incorporated in a model of the Varian CL2300 linear accelerator 6 MV photon beam. As an initial validation of the code, the leaf geometry was visualized by tracing particles through the component module and recording their position each time a leaf boundary was crossed. The leaf dimensions were measured and the leaf material density and interleaf air gap were chosen to match the simulated leaf leakage profiles with film measurements in a solid water phantom. A comparison between Monte Carlo calculations and measurements (diode, radiographic film) was performed for square and irregularly shaped fields incident on flat and homogeneous water phantoms. Results show that Monte Carlo calculations agree with measured dose distributions to within 2% and/or 1 mm except for field size smaller than 1.2 cm diameter where agreement is within 5% due to uncertainties in measured output factors.

  11. Role of oxidative stress and intracellular calcium in nickel carbonate hydroxide-induced sister-chromatid exchange, and alterations in replication index and mitotic index in cultured human peripheral blood lymphocytes

    Energy Technology Data Exchange (ETDEWEB)

    M' Bemba-Meka, Prosper [Universite de Montreal, Human Toxicology Research Group (TOXHUM), Department of Environmental and Occupational Health, Main Station, P.O. Box 6128, Montreal, QC (Canada); University of Louisville, Department of Pharmacology and Toxicology, Center for Genetics and Molecular Medicine, Louisville, KY (United States); Lemieux, Nicole [Universite de Montreal, Department of Pathology and Cellular Biology, Faculty of Medicine, Main Station, P.O. Box 6128, Montreal, QC (Canada); Chakrabarti, Saroj K. [Universite de Montreal, Human Toxicology Research Group (TOXHUM), Department of Environmental and Occupational Health, Main Station, P.O. Box 6128, Montreal, QC (Canada)

    2007-02-15

    Human peripheral lymphocytes from whole blood cultures were exposed to either soluble form of nickel carbonate hydroxide (NiCH) (0-60 {mu}M), or of nickel subsulfide (Ni{sub 3}S{sub 2}) (0-120 {mu}M), or of nickel oxide (NiO) (0-120 {mu}M), or nickel sulfate (NiSO{sub 4}) (0-120 {mu}M) for a short duration of 2 h. The treatments occurred 46 h after the beginning of the cultures. The cultures were harvested after a total incubation of 72 h, and sister-chromatid exchange (SCE), replication index (RI), and mitotic index (MI) were measured for each nickel compound. The soluble form of NiCH at 30 {mu}M but those of Ni{sub 3}S{sub 2} and NiO at 120 {mu}M produced significant increase in the SCE per cell compared to the control value, whereas NiSO{sub 4} failed to produce any such significant increase. Except NiSO{sub 4}, the soluble forms of NiCH, Ni{sub 3}S{sub 2}, and NiO produced significant cell-cycle delay (as measured by the inhibition of RI) as well as significant inhibition of the MI at respective similar concentrations as mentioned above. Pretreatment of human blood lymphocytes with catalase (H{sub 2}O{sub 2} scavenger), or superoxide dismutase (superoxide anion scavenger), or dimethylthiourea (hydroxyl radical scavenger), or deferoxamine (iron chelator), or N-acetylcysteine (general antioxidant) inhibited NiCH-induced SCE, and changes in RI and MI. This suggests the participation of oxidative stress involving H{sub 2}O{sub 2}, the superoxide anion radical, the hydroxyl radical, and iron in the NiCH-induced genotoxic responses. Cotreatment of NiCH with either verapamil (inhibitor of intracellular calcium ion ([Ca{sup 2+}]{sub i}) movement through plasma membranes), or dantrolene (inhibitor of [Ca{sup 2+}]{sub i} release from sarcoplasmic reticulum), or BAPTA (Ca{sup 2+} chelator) also inhibited the NiCH-induced responses. These results suggest that [Ca{sup 2+}]{sub i} is also implicated in the genotoxicity of NiCH. Overall these data indicate that various types

  12. What Is Chronic Lymphocytic Leukemia?

    Science.gov (United States)

    ... Chronic Lymphocytic Leukemia (CLL) About Chronic Lymphocytic Leukemia What Is Chronic Lymphocytic Leukemia? Cancer starts when cells ... body, including the lymph nodes, liver, and spleen. What is leukemia? Leukemia is a cancer that starts ...

  13. Alvocidib in Treating Patients With B-Cell Chronic Lymphocytic Leukemia or Small Lymphocytic Lymphoma

    Science.gov (United States)

    2013-07-01

    B-cell Chronic Lymphocytic Leukemia; Contiguous Stage II Small Lymphocytic Lymphoma; Noncontiguous Stage II Small Lymphocytic Lymphoma; Stage I Chronic Lymphocytic Leukemia; Stage I Small Lymphocytic Lymphoma; Stage II Chronic Lymphocytic Leukemia; Stage III Chronic Lymphocytic Leukemia; Stage III Small Lymphocytic Lymphoma; Stage IV Chronic Lymphocytic Leukemia; Stage IV Small Lymphocytic Lymphoma

  14. Aryl hydrocarbon mono-oxygenase activity in human lymphocytes

    Energy Technology Data Exchange (ETDEWEB)

    Griffin, G.D.; Schuresko, D.D.

    1981-06-01

    Aryl hydrocarbon mono-oxygenase (AHM), an enzyme of key importance in metabolism of xenobiotic chemicals such as polynuclear aromatic hydrocarbons (PNA), is present in human lymphocytes. Studies investing the relation of activity of AHM in human lymphocytes to parameters such as disease state, PNA exposure, in vitro mitogen stimulation, etc. have been summarized in this report. Some studies have demonstrated increased AHM activity in lymphocytes from cigarette smokers (compared to nonsmokers), and in lung cancer patients when compared to appropriate control groups. These observations are confused by extreme variability in human lymphocyte AHM activities, such variability arising from factors such as genetic variation in AHM activity, variation in in vitro culture conditions which affect AHM activity, and the problematical relationship of common AHM assays to actual PNA metabolism taking place in lymphocytes. If some of the foregoing problems can be adequately addressed, lymphocyte AHM activity could hold the promise of being a useful biomarker system for human PNA exposure.

  15. Integration Analysis of MicroRNA and mRNA Expression Profiles in Human Peripheral Blood Lymphocytes Cultured in Modeled Microgravity

    Directory of Open Access Journals (Sweden)

    C. Girardi

    2014-01-01

    Full Text Available We analyzed miRNA and mRNA expression profiles in human peripheral blood lymphocytes (PBLs incubated in microgravity condition, simulated by a ground-based rotating wall vessel (RWV bioreactor. Our results show that 42 miRNAs were differentially expressed in MMG-incubated PBLs compared with 1 g incubated ones. Among these, miR-9-5p, miR-9-3p, miR-155-5p, miR-150-3p, and miR-378-3p were the most dysregulated. To improve the detection of functional miRNA-mRNA pairs, we performed gene expression profiles on the same samples assayed for miRNA profiling and we integrated miRNA and mRNA expression data. The functional classification of miRNA-correlated genes evidenced significant enrichment in the biological processes of immune/inflammatory response, signal transduction, regulation of response to stress, regulation of programmed cell death, and regulation of cell proliferation. We identified the correlation of miR-9-3p, miR-155-5p, miR-150-3p, and miR-378-3p expression with that of genes involved in immune/inflammatory response (e.g., IFNG and IL17F, apoptosis (e.g., PDCD4 and PTEN, and cell proliferation (e.g., NKX3-1 and GADD45A. Experimental assays of cell viability and apoptosis induction validated the results obtained by bioinformatics analyses demonstrating that in human PBLs the exposure to reduced gravitational force increases the frequency of apoptosis and decreases cell proliferation.

  16. Suppression of lymphocyte proliferation by parainfluenza virus type 3-infected bovine alveolar macrophages.

    Science.gov (United States)

    Basaraba, R J; Brown, P R; Laegreid, W W; Silflow, R M; Evermann, J F; Leid, R W

    1993-06-01

    Lymphocytes stimulated with concanavalin A (Con A) or antigen in the presence of bovine parainfluenza virus type 3 (PIV-3) infected bovine alveolar macrophages (BAM) or monocytes, had depressed [3H]thymidine incorporation. This failure of lymphocytes to incorporate radiolabel required live virus, was time dependent and was most pronounced when BAM were infected for 48 hr prior to the addition of lymphocytes. The rate of infection of alveolar macrophages and the release of infectious virus into culture supernatants paralleled suppression of lymphocyte mitogenesis by PIV-3. However, the peak titre of exogenous, live or inactivated virus was not suppressive when added to lymphocyte macrophage cultures just prior to Con A stimulation. Neither the loss of viable alveolar macrophages nor a shift in antigen or mitogen dose response in virally infected cultures could account for the deficit in [3H]thymidine incorporation by lymphocytes. Despite the presence of lymphocyte-associated virus antigen detected by direct immunofluorescence, no increase in PIV-3 titre above baseline was seen from infected lymphocytes, irrespective of mitogen stimulation. Likewise, lymphocytes did not contribute to the extracellular virus pool in lymphocyte-macrophage cultures as the increases in viral titre above basal levels in supernatants were equal to levels released by macrophages alone. The expression of viral antigen on lymphocytes stimulated in the presence of PIV-3-infected BAM suggests a non-productive or abortive infection of lymphocytes mediated through contact with infected macrophages.

  17. SU-E-T-479: IMRT Plan Recalculation in Patient Based On Dynalog Data and the Effect of a Single Failing MLC Motor

    Energy Technology Data Exchange (ETDEWEB)

    Morcos, M [Vantage Oncology, San Bernardino, CA (United States); Mitrou, E [Centre Hospitalier de l’Universite de Montreal, Montreal, QC (Canada)

    2015-06-15

    Purpose: Using Linac dynamic logs (Dynalogs) we evaluate the impact of a single failing MLC motor on the deliverability of an IMRT plan by assessing the recalculated dose volume histograms (DVHs) taking the delivered MLC positions and beam hold-offs into consideration. Methods: This is a retrospective study based on a deteriorating MLC motor (leaf 36B) which was observed to be failing via Dynalog analysis. To investigate further, Eclipse-importable MLC files were generated from Dynalogs to recalculate the actual delivered dose and to assess the clinical impact through DVHs. All deliveries were performed on a Varian 21EX linear accelerator equipped with Millennium-120 MLC. The analysis of Dynalog files and subsequent conversion to Eclipse-importable MLC files were all performed by in-house programming in Python. Effects on plan DVH are presented in the following section on a particular brain-IMRT plan which was delivered with a failing MLC motor which was then replaced. Results: Global max dose increased by 13.5%, max dose to the brainstem PRV increased by 8.2%, max dose to the optic chiasm increased by 7.6%, max dose to optic nerve increased by 8.8% and the mean dose to the PTV increased by 7.9% when comparing the original plan to the fraction with the failing MLC motor. The reason the dose increased was due to the failure being on the B-bank which is the lagging side on a sliding window delivery, therefore any failures on this side will cause an over-irradiation as the B-bank leaves struggles to keep the window from growing. Conclusion: Our findings suggest that a single failing MLC motor may jeopardize the entire delivery. This may be due to the bad MLC motor drawing too much current causing all MLCs on the same bank to underperform. This hypothesis will be investigated in a future study.

  18. Analysis of the impact of uncertainties in the movement of a MLC to perform SBRT of prostate by IMAT; Analisis del impacto de las incertidumbres en el movimiento de un MLC al effectuar SBRT de prostata mediante IMAT

    Energy Technology Data Exchange (ETDEWEB)

    Reynes llomport, G.; Sancho Kolster, I.; Blas Pinol, R. de; Modolell Farre, I.; Picon Olmos, C.

    2013-07-01

    The present work shows, dosimetric level, how the technique of SBRT (Steretactic Body Radiation Therapy), studied can be administered safely, using accepted tolerances in the systematic uncertainty of MLC truck, accepted in the rest of the used techniques, although they suggest approaching the stock level tolerances for a better control of the clinical response in this type of treatment. (Author)

  19. Culture and identification of mouse bone marrow derived DCs that induce T lymphocyte proliferation%小鼠骨髓来源树突状细胞培养鉴定和诱导T淋巴细胞增殖

    Institute of Scientific and Technical Information of China (English)

    吴昆; 拜合提亚·阿扎提; 王文光; 安尼瓦尔·牙生; 王玉杰

    2011-01-01

    Objective:To establish a method of inducing and culturing dendritic cells from mouse bone marrow in vitro,and observe their biological feature and function of stimulating T lymphocytes proliferate in different stages. Methods: Mononuclear cells isolated from mouse bone marrow were induced into dendritic cells by being cultured with GM-CSF and IL-4,and then examined from aspects of morphology,phenotype and function. Results:Modality of mononuclear cells diversified after being cultured in CM-CSF and IL-4 for 3 days,and growthing clustered-liked. When culturing for S days,typical morphology with dendritic processes can be observed. A large number of morphologically typical dendritic cells were observed after culturing for 8 days,under scan electron microscope. DCs displayed typical morphology with dendritic processes and high expressed specific marker of bone marrow derived DC-CD11c and costimulatory molecules of CD40,CD80,CD86 and MHC-Ⅱ. Mature BMDC could stimulate syngenic and allogenic mixed lymphocyte proliferation. Conclusion: A large number of dendritic cells can be generated by culturing the mononuclear cells derived from mouse bone marrow in vitro, and thus will lay a foundation for future research in producing the anti-tumor vaccine.%目的:建立一种体外诱导培养小鼠树突状细胞(DCs)的方法,并观察其不同生长阶段生物学形态及刺激淋巴细胞增殖的能力。方法:联合应用粒细胞-巨噬细胞集落刺激因子(GM-CSF)、白细胞介素-4(IL-4)诱导培养小鼠骨髓单个核细胞向DCs分化,并从形态学、表型及功能方面对其加以检测。结果:用GM-CSF和IL-4诱导培养小鼠骨髓来源的单核细胞,3 d后可见细胞形态改变,并呈集簇生长,培养至5 d可见典型的树突状突起,体外诱导培养8 d后获得大量成熟的DCs,扫描电镜可见典型的树突状细胞形态,高表达骨髓来源DCs特异性标记CD11c和共刺激分子CD40、CD80、CD86和MHC-Ⅱ

  20. Lymphocytes in patients with psoriasis promote proliferation of keratinocytes

    Institute of Scientific and Technical Information of China (English)

    DENG An-mei; ZHONG Ren-qian; CHEN Sun-xiao; ZHOU Ye; KONG Xian-tao

    2002-01-01

    Objective: To analyze the effect of lymphocytes on proliferation of keratinocytes in patients with psoriasis. Methods: Lymphocytes in lesion and peripheral blood were isolated and amplified, then cultured together with normal keratinocytes. By MTT method, the living cells were quantified in the mixed culture.Results: Compared with normal controls, lymphocytes from lesion and peripheral blood of psoriasis both promote the proliferation of keratinocytes (P<0. 01 and P<0. 05 respectively). The concentrations of IL-2 and IFN-γ in the mixture of lesion lymphocytes and keratinocytes were significantly higher than that of controls.Tripterygium glycosides inhibited this promotion. Conclusion: Lymphocytes in patients with psoriasis (mainly Thl cell) play an important role in proliferation of keratinocytes. This psoriasis cell model is useful for studies on signal transduction in psoriasis.

  1. Lymphocytic colitis: A clue to bacterial etiology

    Institute of Scientific and Technical Information of China (English)

    Thanaa EA Helal; Naglaa S Ahmed; Osama Abo El Fotoh

    2005-01-01

    AIM: To find out the role of bacteria as a possible etiological factor in lymphocytic colitis.METHODS: Twenty patients with histopathological diagnosis of lymphocytic colitis and 10 normal controls were included in this study. Colonoscopic biopsies were obtained from three sites (hepatic and splenic flexures and rectosigmoid region). Each biopsy was divided into two parts. A fresh part was incubated on special cultures for bacterial growth. The other part was used for the preparation of histologic tissue sections that were examined for the presence of bacteria with the help of Giemsa stain.RESULTS: Culture of tissue biopsies revealed bacterial growth in 18 out of 20 patients with lymphocytic colitis mostly Escherichia coli(14/18), which was found in all rectosigmoid specimens (14/14), but only in 8/14 and 6/14 of splenic and hepatic flexure specimens respectively. In two of these cases, E coliwas associated with proteus. Proteus was found only in one case, Klebsiella in two cases, and Staphylococcus aureus in one case. In the control group, only 2 out of 10 controls showed the growth of E coliin their biopsy cultures.Histopathology showed rod-shaped bacilli in the tissue sections of 12 out of 14 cases with positive E coliin their specimen's culture. None of the controls showed these bacteria in histopathological sections.CONCLUSION: This preliminary study reports an association between E coliand lymphocytic colitis, based on histological and culture observations. Serotyping and molecular studies are in process to assess the role of E coliin the pathogenesis of lymphocytic colitis.

  2. Apoptosis of matured T lymphocytes induced by mouse sertoli cells in cocultures in vitro

    Institute of Scientific and Technical Information of China (English)

    LIU Yang; LIN Zi-hao; ZHU Xiao-hai; LIU Shan-rong

    2001-01-01

    Objective: To study whether mouse sertoli cells can induce the apoptosis of matured T lymphocytes in cocultures in vitro. Methods: With TUNEL, DNA electrophoresis, eleetro-mierography and flow cytometry, we examined the apoptosis and its rates of mouse matured T lymphocytes in control group (T lymphocytes only), group A (T lymphocytes + culture medium of sertoli cells), group B (T lymphocytes + sertoli cells). Results: Under electro-micrography, chromatin condensation, karyopyknosis, karyorhexis and apoptotic body were observed in some T lymphocytes in 3 groups; some nucleuses were stained dark blue with TUNEL; a typical DNA ladder was found with DNA electrophoresis. The apoptotic rates of T lymphocytes in group A and B were significantly higher than that in control group (P<0.01). The apoptotic rate of T lymphocytes in group B was significantly higher than that in group A (P<0.01). Conclusion: In coculture condition in vitro,mouse sertoli cells can induce the apoptosis of matured T lymphocytes.

  3. SU-E-T-633: Dose Differences in Lung Cancer SBRT: The Influences of MLC Width

    Energy Technology Data Exchange (ETDEWEB)

    Chen, J; Yin, Y [Shandong Cancer Hospital, Jinan, Shandong (China)

    2014-06-15

    Purpose: The aim is to compare the plan dose distribution of lung SBRT with MLCs in different width. Methods: Cases with phase INSCLC were enrolled. 9 cases were undergone 4D-CT scanning in the supine position with both arms raised. 3D-CT images without IV contrast were afterwards acquired with 3mm thickness and used for dose calculations. ITV was generated by using the inspiration and expiration images. The ITV can be expanded by geometric set-up uncertainty (5 mm) to generate the PTV. All chest normal tissues including chest wall were contoured by doctors. A total dose of 55 Gy will be given in 5 fractions within 10–14 days with an inter fraction interval of 2–3 days. Guided by the RTOG trial 3502 protocol, 11–13 non-coplanar fields with 6MV photon were arranged. Three types of MLCs with width of 3mm, 5mm and 10mm at isocenter position, were used separately to generate a CRT plan for each case. Monte Carlo algorithm was applied to dose calculation. All plans were adjusted as possible to meet the dose constraints. Dose-volume parameters from plans as followed were compared and analysized: PTV V55Gy, COMPTV D70% (70% of normalization dose), volume A (body minus PTV), and R100% and R50% (the ratio of x% of prescription dose isoline volume to PTV volume). Results: MLCs, 3mm and 5mm wide, played the identical roles on dosimetry of the plans, excluding the parameter volume A (p<0.05). On the contrary, MLC with width of 10mm was significantly inferior to the other two types on most parameters (p<0.05). For R50%, all types contributed equally (p>0.05). Conclusion: For lung cancer SBRT, MLC width had influence to dosimetry, especially in irradiation area. Small size MLC, e.g. 3mm and 5mm, are helpful to generate a high quality treatment plan, which could meet the strict criteria for targets and OAR.

  4. Impact of MLC leaf position errors on simple and complex IMRT plans for head and neck cancer

    Energy Technology Data Exchange (ETDEWEB)

    Mu, G; Ludlum, E; Xia, P [Department of Radiation Oncology, University of California, San Francisco, CA (United States)

    2008-01-07

    The dosimetric impact of random and systematic multi-leaf collimator (MLC) leaf position errors is relatively unknown for head and neck intensity-modulated radiotherapy (IMRT) patients. In this report we studied 17 head and neck IMRT patients, including 12 treated with simple plans (<50 segments) and 5 treated with complex plans (>100 segments). Random errors (-2 to +2 mm) and systematic errors ({+-}0.5 mm and {+-}1 mm) in MLC leaf positions were introduced into the clinical plans and the resultant dose distributions were analyzed based on defined endpoint doses. The dosimetric effect was insignificant for random MLC leaf position errors up to 2 mm for both simple and complex plans. However, for systematic MLC leaf position errors, we found significant dosimetric differences between the simple and complex IMRT plans. For 1 mm systematic error, the average changes in D{sub 95%} were 4% in simple plans versus 8% in complex plans. The average changes in D{sub 0.1cc} of the spinal cord and brain stem were 4% in simple plans versus 12% in complex plans. The average changes in parotid glands were 9% in simple plans versus 13% for the complex plans. Overall, simple IMRT plans are less sensitive to leaf position errors than complex IMRT plans.

  5. SU-E-T-122: Dosimetric Comparison Between Cone, HDMLC and MicroMLC for the Treatment of Trigeminal Neuralgia

    Energy Technology Data Exchange (ETDEWEB)

    Vacca, N; Caussa, L; Filipuzzi, M; Garrigo, E; Venencia, C [Instituto Privado de Radioterapia - Fundacion Marie Curie, Cordoba (Argentina)

    2014-06-01

    Purpose: The purpose of this work was to evaluate the dosimetric characteristics of three collimation systems, 5mm circular cone (Brainlab) and square fields of 5mm with HDMLC (Varian) and microMLC Moduleaf, Siemens) for trigeminal neuralgia treatment. Methods: A TPS Iplan v4.5 BrainLAB was used to do treatment plans for each collimations system in a square solid water phantom with isocenter at 5cm depth. Single field and treatment plan including 11 arcs with fix field and 100° gantry range was made for each collimation systems. EBT3 films were positioned at isocenter in a coronal plane to measured dose distribution for all geometries. Films were digitized with a Vidar DosimetryPro Red scanner with a resolution of 89dpi and RIT113v6.1 software was used for analysis. Penumbra region (80%–20%), FWHM and dose percentage at 5mm and 10mm from CAX were determined. All profiles were normalized at CAX. Results: For single beam the penumbra (FWHM) was 1.5mm (5.3mm) for the cone, 1.9mm (5.5mm) for HDMLC and 1.8mm (5.4mm) for the microMLC. Dose percentage at 5mm was 6.9% for cone, 12.5% for HDMLC and 8.7% for the microMLC. For treatment plan the penumbra (FWHM) was 2.58mm (5.47mm) for the cone, 2.8mm (5.84mm) for HDMLC and 2.58mm (6.09mm) for the microMLC. Dose perecentage at 5mm was 13.1% for cone, 16.1% for HDMLC, 15.2% for the microMLC. Conclusion: The cone has a dose falloff larger than the microMLC and HDMLC, by its reduced penumbra, this translates into better protection of surrounding healthy tissue, however, the microMLC and HDMLC have similar accuracy to cone.

  6. Beamlet based direct aperture optimization for MERT using a photon MLC

    Energy Technology Data Exchange (ETDEWEB)

    Henzen, D., E-mail: henzen@ams.unibe.ch; Manser, P.; Frei, D.; Volken, W.; Born, E. J.; Joosten, A.; Lössl, K.; Aebersold, D. M.; Chatelain, C.; Fix, M. K. [Division of Medical Radiation Physics and Department of Radiation Oncology, Inselspital, Bern University Hospital, and University of Bern, Berne CH-3010 (Switzerland); Neuenschwander, H. [Clinic for Radiation-Oncology, Lindenhofspital, Berne CH-3012 (Switzerland); Stampanoni, M. F. M. [Institute for Biomedical Engineering, ETH Zürich and Paul Scherrer Institut, Villigen CH-5234 (Switzerland)

    2014-12-15

    Purpose: A beamlet based direct aperture optimization (DAO) for modulated electron radiotherapy (MERT) using photon multileaf collimator (pMLC) shaped electron fields is developed and investigated. Methods: The Swiss Monte Carlo Plan (SMCP) allows the calculation of dose distributions for pMLC shaped electron beams. SMCP is interfaced with the Eclipse TPS (Varian Medical Systems, Palo Alto, CA) which can thus be included into the inverse treatment planning process for MERT. This process starts with the import of a CT-scan into Eclipse, the contouring of the target and the organs at risk (OARs), and the choice of the initial electron beam directions. For each electron beam, the number of apertures, their energy, and initial shape are defined. Furthermore, the DAO requires dose–volume constraints for the structures contoured. In order to carry out the DAO efficiently, the initial electron beams are divided into a grid of beamlets. For each of those, the dose distribution is precalculated using a modified electron beam model, resulting in a dose list for each beamlet and energy. Then the DAO is carried out, leading to a set of optimal apertures and corresponding weights. These optimal apertures are now converted into pMLC shaped segments and the dose calculation for each segment is performed. For these dose distributions, a weight optimization process is launched in order to minimize the differences between the dose distribution using the optimal apertures and the pMLC segments. Finally, a deliverable dose distribution for the MERT plan is obtained and loaded back into Eclipse for evaluation. For an idealized water phantom geometry, a MERT treatment plan is created and compared to the plan obtained using a previously developed forward planning strategy. Further, MERT treatment plans for three clinical situations (breast, chest wall, and parotid metastasis of a squamous cell skin carcinoma) are created using the developed inverse planning strategy. The MERT plans are

  7. Dosimetric evaluation of photon dose calculation under jaw and MLC shielding

    Energy Technology Data Exchange (ETDEWEB)

    Fogliata, A.; Clivio, A.; Vanetti, E.; Nicolini, G.; Belosi, M. F.; Cozzi, L. [Oncology Institute of Southern Switzerland, Medical Physics Unit, Bellinzona 6500 (Switzerland)

    2013-10-15

    Purpose: The accuracy of photon dose calculation algorithms in out-of-field regions is often neglected, despite its importance for organs at risk and peripheral dose evaluation. The present work has assessed this for the anisotropic analytical algorithm (AAA) and the Acuros-XB algorithms implemented in the Eclipse treatment planning system. Specifically, the regions shielded by the jaw, or the MLC, or both MLC and jaw for flattened and unflattened beams have been studied.Methods: The accuracy in out-of-field dose under different conditions was studied for two different algorithms. Measured depth doses out of the field, for different field sizes and various distances from the beam edge were compared with the corresponding AAA and Acuros-XB calculations in water. Four volumetric modulated arc therapy plans (in the RapidArc form) were optimized in a water equivalent phantom, PTW Octavius, to obtain a region always shielded by the MLC (or MLC and jaw) during the delivery. Doses to different points located in the shielded region and in a target-like structure were measured with an ion chamber, and results were compared with the AAA and Acuros-XB calculations. Photon beams of 6 and 10 MV, flattened and unflattened were used for the tests.Results: Good agreement between calculated and measured depth doses was found using both algorithms for all points measured at depth greater than 3 cm. The mean dose differences (±1SD) were −8%± 16%, −3%± 15%, −16%± 18%, and −9%± 16% for measurements vs AAA calculations and −10%± 14%, −5%± 12%, −19%± 17%, and −13%± 14% for Acuros-XB, for 6X, 6 flattening-filter free (FFF), 10X, and 10FFF beams, respectively. The same figures for dose differences relative to the open beam central axis dose were: −0.1%± 0.3%, 0.0%± 0.4%, −0.3%± 0.3%, and −0.1%± 0.3% for AAA and −0.2%± 0.4%, −0.1%± 0.4%, −0.5%± 0.5%, and −0.3%± 0.4% for Acuros-XB. Buildup dose was overestimated with AAA, while Acuros-XB gave

  8. Dosimetric evaluation of photon dose calculation under jaw and MLC shielding.

    Science.gov (United States)

    Fogliata, A; Clivio, A; Vanetti, E; Nicolini, G; Belosi, M F; Cozzi, L

    2013-10-01

    The accuracy of photon dose calculation algorithms in out-of-field regions is often neglected, despite its importance for organs at risk and peripheral dose evaluation. The present work has assessed this for the anisotropic analytical algorithm (AAA) and the Acuros-XB algorithms implemented in the Eclipse treatment planning system. Specifically, the regions shielded by the jaw, or the MLC, or both MLC and jaw for flattened and unflattened beams have been studied. The accuracy in out-of-field dose under different conditions was studied for two different algorithms. Measured depth doses out of the field, for different field sizes and various distances from the beam edge were compared with the corresponding AAA and Acuros-XB calculations in water. Four volumetric modulated arc therapy plans (in the RapidArc form) were optimized in a water equivalent phantom, PTW Octavius, to obtain a region always shielded by the MLC (or MLC and jaw) during the delivery. Doses to different points located in the shielded region and in a target-like structure were measured with an ion chamber, and results were compared with the AAA and Acuros-XB calculations. Photon beams of 6 and 10 MV, flattened and unflattened were used for the tests. Good agreement between calculated and measured depth doses was found using both algorithms for all points measured at depth greater than 3 cm. The mean dose differences (± 1SD) were -8% ± 16%, -3% ± 15%, -16% ± 18%, and -9% ± 16% for measurements vs AAA calculations and -10% ± 14%, -5% ± 12%, -19% ± 17%, and -13% ± 14% for Acuros-XB, for 6X, 6 flattening-filter free (FFF), 10X, and 10FFF beams, respectively. The same figures for dose differences relative to the open beam central axis dose were: -0.1% ± 0.3%, 0.0% ± 0.4%, -0.3% ± 0.3%, and -0.1% ± 0.3% for AAA and -0.2% ± 0.4%, -0.1% ± 0.4%, -0.5% ± 0.5%, and -0.3% ± 0.4% for Acuros-XB. Buildup dose was overestimated with AAA, while Acuros-XB gave results more consistent with

  9. MLC2450无烟煤立磨的开发设计

    Institute of Scientific and Technical Information of China (English)

    谢伟安

    2014-01-01

    MLC2450无烟煤立磨是北方重工集团有限公司水泥设备分公司在MPF煤立磨、MLK矿渣立磨成功经验的基础上,从无烟煤的特性入手,自行开发设计的高细煤粉粉磨设备。文章简要介绍了该立磨的结构组成和工作原理,详细分析了该磨的技术特点和性能优势。粉磨无烟煤具有能耗低、磨损小、运行平稳等特点。

  10. Commissioning, evaluation, quality assurance and clinical application of a virtual micro MLC technique.

    Science.gov (United States)

    Woo, M; Charland, P; Kim, B; Nico, A

    2003-02-01

    Multileaf collimators (MLCs) are a valuable tool in modern radiation therapy, offering flexible and convenient field shaping. One disadvantage, however, is the undulation of the dose distribution at the edge shaped by the leaves due to the finite leaf width. An attempt to reduce the effect of this undulation is the objective of the commercial linear accelerator package HD270, which incorporates three-dimensional couch translation together with leaf adjustment to emulate finer leaf widths. In this paper we report on the commissioning and evaluation of this feature, together with the development of a process for quality assurance, as well as description of a clinical application of this technique. It is concluded that this technique could be applied reliably to situations currently utilizing MLC for shielding, with little added cost in treatment time, provided that a comprehensive quality assurance program is in place to monitor the performance of this complicated procedure.

  11. Acute Lymphocytic Leukemia

    Science.gov (United States)

    ... for information in your local library and on the Internet. Good sources include the National Cancer Institute, the ... mayoclinic.org/diseases-conditions/acute-lymphocytic-leukemia/basics/definition/CON-20042915 . Mayo Clinic Footer Legal Conditions and ...

  12. Evaluation of MLC leaf positioning accuracy for static and dynamic IMRT treatments using DAVID in vivo dosimetric system.

    Science.gov (United States)

    Karagoz, Gulay; Zorlu, Faruk; Yeginer, Mete; Yildiz, Demet; Ozyigit, Gokhan

    2016-03-08

    Accuracy and precision of leaf positioning in multileaf collimators (MLCs) are significant factors for the accuracy of IMRT treatments. This study aimed to inves-tigate the accuracy and repeatability of the MLC leaf positioning via the DAVID invivo dosimetric system for dynamic and static MLC systems. The DAVID system was designed as multiwire transmission ionization chamber which is placed in accessory holder of linear accelerators. Each wire of DAVID system corresponds to a MLC leaf-pair to verify the leaf positioning accuracy during IMRT treatment and QA. In this study, verifications of IMRT plans of five head and neck (H&N) and five prostate patients treated in a Varian DHX linear accelerator with 80-leaf MLC were performed using DAVID system. Before DAVID-based dosimetry, Electronics Portal Imaging Device (EPID) and PTW 2D ARRAY dosimetry system were used for 2D verification of each plan. The measurements taken by DAVID system in the first day of the treatments were used as reference for the following measurements taken over the next four weeks. The deviations in leaf positioning were evaluated by "Total Deviation (TD)" parameter calculated by DAVID software. The delivered IMRT plans were originally prepared using dynamic MLC method. The same plans were subsequently calculated based on static MLC method with three different intensity levels of five (IL5), 10 (IL10) and 20 (IL20) in order to compare the performances of MLC leaf positioning repeatability for dynamic and static IMRT plans. The leaf positioning accuracy is also evaluated by analyzing DynaLog files based on error histograms and root mean square (RMS) errors of leaf pairs' positions. Moreover, a correlation analysis between simultaneously taken DAVID and EPID measurements and DynaLog file recordings was subsequently performed. In the analysis of DAVID outputs, the overall deviations of dynamic MLC-based IMRT calculated from the deviations of the four weeks were found as 0.55% ± 0.57% and 1.48% ± 0

  13. Mutagenic biomonitoring of pirethroid insecticides in human lymphocyte cultures: use of micronuclei as biomarkers and recovery by Rosa canina extracts of mutagenic effects.

    Science.gov (United States)

    Kasimoglu, Caner; Uysal, Handan

    2015-05-01

    Insecticides are used to control pests. Cypermethrin and fenvalerate are widely used pirethroid insecticides in the world. Rosa canina L. (Rosaceae) is used as a traditional medicinal plant against viral infections and disorders of the kidneys and urinary tract due to its high vitamin C level. The genotoxic effects of cypermethrin and fenvalerate were examined with the micronucleus (MN) test. Then, we determined the ability of the water (RC(wtr)) and ethanol (RC(eta)) extracts of rosehip (R. canina) to overcome the possible genotoxic effects of the insecticides. Preliminary studies determined that the application concentrations were 20, 30, 40, and 50 ppm for cypermethrin, 25, 50, 75, and 100 ppm for fenvalerate, and 100 ppm for rosehip extracts. DMSO (dimethyl sulphoxide) (1%) and 1 mM EMS (ethyl methanesulfonate) were used as negative and positive control groups, respectively. The application groups belonging to insecticides and plant extracts were added to culture tubes including chromosome B medium and peripheral blood for MN test. The MN frequencies were found 0.725 in the negative control group, 2.700 in the positive control groups, 1.275 in the highest application group of cypermethrin, and 1.600 in the highest application group of fenvalerate. The MN frequencies in cypermethrin + RC(wtr), cypermethrin + RC(eta), fenvalerate + RC(wtr), and fenvalerate + RC(eta) application groups were, respectively, determined as 1.000, 1.075, 1.225, and 1.275. According to the results, cypermethrin and fenvalerate have genotoxic effects, the water and ethanol extracts of rosehip reduced the genotoxicity of the both insecticides.

  14. The efflux pump MlcE from the Penicillium solitum compactin biosynthetic gene cluster increases Saccharomyces cerevisiae resistance to natural statins

    DEFF Research Database (Denmark)

    Ley, Ana; Frandsen, Rasmus John Normand

    in yeast, which would overcome the undesirable effects of statins. One possible mechanism is an active export of statins, a mechanism that does not just provide the resistance but can also significantly ease the purification of the produced compounds. In order to establish export of statins from yeast we...... integrated a putative efflux pump-encoding gene mlcE from the P. solitum compactin biosynthetic gene cluster into S. cerevisiae genome. The resulting strain was tested for susceptibility to statins by growing the strain on media containing statins. The constructed strain showed an increased resistance...... to both natural statins (compactin and lovastatin), and also to a semi-synthetic statin simvastatin, when compared to the wild type strain. Expression of a mRFP-tagged MlcE show that MlcE is localized in the yeast plasma membrane. In conclusion we provide evidence indicating that MlcE is a transmembrane...

  15. Chemokines, lymphocytes, and HIV

    Directory of Open Access Journals (Sweden)

    Farber J.M.

    1998-01-01

    Full Text Available Chemokines are members of a family of more than 30 human cytokines whose best-described activities are as chemotactic factors for leukocytes and that are presumed to be important in leukocyte recruitment and trafficking. While many chemokines can act on lymphocytes, the roles of chemokines and their receptors in lymphocyte biology are poorly understood. The recent discoveries that chemokines can suppress infection by HIV-1 and that chemokine receptors serve, along with CD4, as obligate co-receptors for HIV-1 entry have lent urgency to studies on the relationships between chemokines and lymphocytes. My laboratory has characterized Mig and Crg-2/IP-10, chemokines that are induced by IFN-g and that specifically target lymphocytes, particularly activated T cells. We have demonstrated that the genes for these chemokines are widely expressed during experimental infections in mice with protozoan and viral pathogens, but that the patterns of mig and crg-2 expression differed, suggesting non-redundant roles in vivo. Our related studies to identify new chemokine receptors from activated lymphocytes resulted in the cloning of STRL22 and STRL33. We and others have shown that STRL22 is a receptor for the CC chemokine MIP-3a, and STRL22 has been re-named CCR6. Although STRL33 remains an orphan receptor, we have shown that it can function as a co-receptor for HIV-1 envelope glycoproteins, and that it is active with a broader range of HIV-1 envelope glycoproteins than the major co-receptors described to date. The ability of STRL33 to function with a wide variety of envelope glycoproteins may become particularly important if therapies are instituted to block other specific co-receptors. We presume that investigations into the roles of chemokines and their receptors in lymphocyte biology will provide information important for understanding the pathogenesis of AIDS and for manipulating immune and inflammatory responses for clinical benefit

  16. SU-C-BRB-04: Characteristics and Performance Evaluation of the First Commercial MLC for a Robotic Delivery System

    Energy Technology Data Exchange (ETDEWEB)

    Fuerweger, C [Erasmus MC Cancer Institute, Rotterdam (Netherlands); European Cyberknife Center Munich, Munich, DE (Germany); Prins, P; Coskan, H; Heijmen, B [Erasmus MC Cancer Institute, Rotterdam (Netherlands)

    2015-06-15

    Purpose: To assess characteristics and performance of the “Incise™” MLC (41 leaf pairs, 2.5mm width, FFF linac) mounted on the robotic SRS/SBRT platform “CyberKnife M6™” in a pre-clinical 5 months (11/2014–03/2015) test period. Methods: Beam properties were measured with unshielded diodes and EBT3 film. The CyberKnife workspace for MLC was analyzed by transforming robot node coordinates (cranial / body paths) into Euler geometry. Bayouth tests for leaf / bank position accuracy were performed in standard (A/P) and clinically relevant non-standard positions, before and after exercising the MLC for 10+ minutes. Total system and delivery accuracy were assessed in End-to-End tests and dosimetric verification of exemplary plans. Stability over time was evaluated in Picket-Fence-and adapted Winston-Lutz-tests (AQA) for different collimator angles. Results: Penumbrae (80–20%, with 100%=2*dose at inflection point; SAD 80cm; 10cm depth) parallel / perpendicular to leaf motion were 2.87/2.64mm for the smallest (0×76×0.75cm{sup 2}) and 5.34/4.94mm for the largest (9.76×9.75cm{sup 2}) square field. MLC circular field penumbrae exceeded fixed cones by 10–20% (e.g. 60mm: 4.0 vs. 3.6mm; 20mm: 3.6 vs. 2.9mm). Interleaf leakage was <0.5%. Clinically accessible workspace with MLC covered (non-coplanar) gantry angles of [-113°;+112°] (cranial) and [-108°;+102°] (body), and collimator angles of [-100°;+107°] (cranial) and [-91°;+100°] (body). Average leaf position offsets were ≤0.2mm in 14 standard A/P Bayouth tests and ≤0.6mm in 8 non-standard direction tests. Pre-test MLC exercise increased jaggedness (range ±0.3mm vs. ±0.5mm) and allowed to identify one malfunctioning leaf motor. Total system accuracy with MLC was 0.39±0.06mm in 6 End-to-End tests. Picket-Fence and AQA showed no adverse trends during the test period. Conclusion: The Incise™ MLC for CyberKnife M6™ displayed high accuracy and mechanical stability over the test period. The

  17. Technical Note: Motion-perturbation method applied to dosimetry of dynamic MLC target tracking—A proof-of-concept

    Energy Technology Data Exchange (ETDEWEB)

    Feygelman, Vladimir, E-mail: vladimir.feygelman@moffitt.org; Tonner, Brian; Hunt, Dylan; Zhang, Geoffrey; Moros, Eduardo [Department of Radiation Oncology, Moffitt Cancer Center, Tampa, Florida 33612 (United States); Stambaugh, Cassandra [Department of Physics, University of South Florida, Tampa, Florida 33612 (United States); Nelms, Benjamin E. [Canis Lupus LLC, Merrimac, Wisconsin 53561 (United States)

    2015-11-15

    Purpose: Previous studies show that dose to a moving target can be estimated using 4D measurement-guided dose reconstruction based on a process called virtual motion simulation, or VMS. A potential extension of VMS is to estimate dose during dynamic multileaf collimator (MLC)-tracking treatments. The authors introduce a modified VMS method and quantify its performance as proof-of-concept for tracking applications. Methods: Direct measurements with a moving biplanar diode array were used to verify accuracy of the VMS dose estimates. A tracking environment for variably sized circular MLC apertures was simulated by sending preprogrammed control points to the MLC while simultaneously moving the accelerator treatment table. Sensitivity of the method to simulated tracking latency (0–700 ms) was also studied. Potential applicability of VMS to fast changing beam apertures was evaluated by modeling, based on the demonstrated dependence of the cumulative dose on the temporal dose gradient. Results: When physical and virtual latencies were matched, the agreement rates (2% global/2 mm gamma) between the VMS and the biplanar dosimeter were above 96%. When compared to their own reference dose (0 induced latency), the agreement rates for VMS and biplanar array track closely up to 200 ms of induced latency with 10% low-dose cutoff threshold and 300 ms with 50% cutoff. Time-resolved measurements suggest that even in the modulated beams, the error in the cumulative dose introduced by the 200 ms VMS time resolution is not likely to exceed 0.5%. Conclusions: Based on current results and prior benchmarks of VMS accuracy, the authors postulate that this approach should be applicable to any MLC-tracking treatments where leaf speeds do not exceed those of the current Varian accelerators.

  18. Technical Note: Motion-perturbation method applied to dosimetry of dynamic MLC target tracking--A proof-of-concept.

    Science.gov (United States)

    Feygelman, Vladimir; Tonner, Brian; Stambaugh, Cassandra; Hunt, Dylan; Zhang, Geoffrey; Moros, Eduardo; Nelms, Benjamin E

    2015-11-01

    Previous studies show that dose to a moving target can be estimated using 4D measurement-guided dose reconstruction based on a process called virtual motion simulation, or VMS. A potential extension of VMS is to estimate dose during dynamic multileaf collimator (MLC)-tracking treatments. The authors introduce a modified VMS method and quantify its performance as proof-of-concept for tracking applications. Direct measurements with a moving biplanar diode array were used to verify accuracy of the VMS dose estimates. A tracking environment for variably sized circular MLC apertures was simulated by sending preprogrammed control points to the MLC while simultaneously moving the accelerator treatment table. Sensitivity of the method to simulated tracking latency (0-700 ms) was also studied. Potential applicability of VMS to fast changing beam apertures was evaluated by modeling, based on the demonstrated dependence of the cumulative dose on the temporal dose gradient. When physical and virtual latencies were matched, the agreement rates (2% global/2 mm gamma) between the VMS and the biplanar dosimeter were above 96%. When compared to their own reference dose (0 induced latency), the agreement rates for VMS and biplanar array track closely up to 200 ms of induced latency with 10% low-dose cutoff threshold and 300 ms with 50% cutoff. Time-resolved measurements suggest that even in the modulated beams, the error in the cumulative dose introduced by the 200 ms VMS time resolution is not likely to exceed 0.5%. Based on current results and prior benchmarks of VMS accuracy, the authors postulate that this approach should be applicable to any MLC-tracking treatments where leaf speeds do not exceed those of the current Varian accelerators.

  19. A dosimetric evaluation of the Eclipse AAA algorithm and Millennium 120 MLC for cranial intensity-modulated radiosurgery.

    Science.gov (United States)

    Calvo Ortega, Juan Francisco; Moragues, Sandra; Pozo, Miquel; José, Sol San; Puertas, Enrique; Fernández, Jaime; Casals, Joan

    2014-01-01

    The aim of this study is to assess the accuracy of a convolution-based algorithm (anisotropic analytical algorithm [AAA]) implemented in the Eclipse planning system for intensity-modulated radiosurgery (IMRS) planning of small cranial targets by using a 5-mm leaf-width multileaf collimator (MLC). Overall, 24 patient-based IMRS plans for cranial lesions of variable size (0.3 to 15.1cc) were planned (Eclipse, AAA, version 10.0.28) using fixed field-based IMRS produced by a Varian linear accelerator equipped with a 120 MLC (5-mm width on central leaves). Plan accuracy was evaluated according to phantom-based measurements performed with radiochromic film (EBT2, ISP, Wayne, NJ). Film 2D dose distributions were performed with the FilmQA Pro software (version 2011, Ashland, OH) by using the triple-channel dosimetry method. Comparison between computed and measured 2D dose distributions was performed using the gamma method (3%/1mm). Performance of the MLC was checked by inspection of the DynaLog files created by the linear accelerator during the delivery of each dynamic field. The absolute difference between the calculated and measured isocenter doses for all the IMRS plans was 2.5% ± 2.1%. The gamma evaluation method resulted in high average passing rates of 98.9% ± 1.4% (red channel) and 98.9% ± 1.5% (blue and green channels). DynaLog file analysis revealed a maximum root mean square error of 0.46mm. According to our results, we conclude that the Eclipse/AAA algorithm provides accurate cranial IMRS dose distributions that may be accurately delivered by a Varian linac equipped with a Millennium 120 MLC.

  20. Immunosuppressant MPA Modulates Tight Junction through Epigenetic Activation of MLCK/MLC-2 Pathway via p38MAPK

    Directory of Open Access Journals (Sweden)

    Niamat Khan

    2015-12-01

    Full Text Available Background: Mycophenolic acid (MPA is an important immunosuppressive drug (ISD prescribed to prevent graft rejection in the organ transplanted patients, however, its use is also associated with adverse side effects like sporadic gastrointestinal (GI disturbances. Recently, we reported the MPA induced tight junctions (TJs deregulation which involves MLCK/MLC-2 pathway. Here, we investigated the global histone acetylation as well as gene-specific chromatin signature of several genes associated with TJs regulation in Caco-2 cells after MPA treatment.Results: The epigenetic analysis shows that MPA treatment increases the global histone acetylation levels as well as the enrichment for transcriptional active histone modification mark (H3K4me3 at promoter regions of p38MAPK, ATF-2, MLCK, and MLC-2. In contrast, the promoter region of occludin was enriched for transcriptional repressive histone modification mark (H3K27me3 after MPA treatment. In line with the chromatin status, MPA treatment increased the expression of p38MAPK, ATF-2, MLCK, and MLC-2 both at transcriptional and translational level, while occludin expression was negatively influenced. Interestingly, the MPA induced gene expression changes and functional properties of Caco-2 cells could be blocked by the inhibition of p38MAPK using a chemical inhibitor (SB203580.Conclusions: Collectively, our results highlight that MPA disrupts the structure of TJs via p38MAPK-dependent activation of MLCK/MLC-2 pathway that results in decreased integrity of Caco-2 monolayer. These results led us to suggest that p38MAPK-mediated lose integrity of epithelial monolayer could be the possible cause of GI disturbance (barrier dysfunction in the intestine, leading to leaky style diarrhea observed in the organ-transplanted patients treated with MPA.

  1. Cyclophosphamide, Alvocidib, and Rituximab in Treating Patients With High Risk B-Cell Chronic Lymphocytic Leukemia or Small Lymphocytic Lymphoma

    Science.gov (United States)

    2015-11-10

    Chronic Lymphocytic Leukemia; Prolymphocytic Leukemia; Recurrent Small Lymphocytic Lymphoma; Refractory Chronic Lymphocytic Leukemia; Stage I Chronic Lymphocytic Leukemia; Stage I Small Lymphocytic Lymphoma; Stage II Chronic Lymphocytic Leukemia; Stage II Small Lymphocytic Lymphoma; Stage III Chronic Lymphocytic Leukemia; Stage III Small Lymphocytic Lymphoma; Stage IV Chronic Lymphocytic Leukemia; Stage IV Small Lymphocytic Lymphoma

  2. T- and B-lymphocyte chimerism in the marmoset

    Energy Technology Data Exchange (ETDEWEB)

    Niblack, G.D.; Kateley, J.R.; Gengozian, N.

    1977-01-01

    Marmosets are natural blood chimeras, this condition resulting from the high frequency of fraternal twinning and the consistent development of placental vascular anastomoses between the two embryos. Identification of chimerism by sex-chromosome analysis of cultured blood lymphocytes provided a means of determining the proportion of chimerism among T and B lymphocytes. Peripheral blood lymphocytes were enriched for T or B cells by filtration through a nylon column (yields >95% T-cells) or inactivation of T lymphocytes by treatment with a goat anti-marmoset thymocyte antiserum in the presence of complement (yields >95% B cells). Mitogenic stimulation of these separated, enriched cell populations yielded metaphase plates which could be scored for percentage male and female cells. Tests on five different blood chimeras showed the T- and B-lymphocyte chimerism to be the same. Stimulation of blood lymphocytes with cells from another species of marmoset in a mixed lymphocyte culture test revealed the chimeric T-cell response (i.e., host and co-twin cells) to be similar to that obtained with a mitogenic lectin. The demonstration of equivalent T- and B-cell chimerism in these animals suggests derivation of these cells from a common stem cell pool and the response of both T-cell populations to an antigenic stimulus in proportions similar to their percentage chimerism suggests complete immunologic tolerance exists in this species for co-twin histocompatibility antigens.

  3. Safety and Efficacy of MLC601 in Iranian Patients after Stroke: A Double-Blind, Placebo-Controlled Clinical Trial

    Directory of Open Access Journals (Sweden)

    A. A. Harandi

    2011-01-01

    Full Text Available Objective. To investigate the safety and efficacy of MLC601 (NeuroAid as a traditional Chinese medicine on motor recovery after ischemic stroke. Methods. This study was a double-blind, placebo-controlled clinical trial on 150 patients with a recent (less than 1 month ischemic stroke. All patients were given either MLC601 (100 patients or placebo (50 patients, 4 capsules 3 times a day, as an add-on to standard stroke treatment for 3 months. Results. Sex, age, elapsed time from stroke onset, and risk factors in the treatment group were not significantly different from placebo group at baseline (P>.05. Repeated measures analysis showed that Fugl-Meyer assessment was significantly higher in the treatment group during 12 weeks after stroke (P<.001. Good tolerability to treatment was shown, and adverse events were mild and transient. Conclusion. MLC601 showed better motor recovery than placebo and was safe on top of standard ischemic stroke medications especially in the severe and moderate cases.

  4. Ameliorative effect of melatonin against increased intestinal permeability in diabetic rats: possible involvement of MLCK-dependent MLC phosphorylation.

    Science.gov (United States)

    Yang, Xiaoping; Zou, Duobing; Tang, Songtao; Fan, Tingting; Su, Huan; Hu, Ruolei; Zhou, Qing; Gui, Shuyu; Zuo, Li; Wang, Yuan

    2016-05-01

    The increased intestinal permeability and functional impairment play an important role in type 2 diabetes (T2D), and melatonin may possess enteroprotection properties. Therefore, we used streptozotocin-induced diabetic rat model to investigate the regulation of intestinal permeability by melatonin. Rats were randomly divided into three groups, including control, diabetes mellitus (DM), and DM rats treated with melatonin. Melatonin was administered (10 mg/kg/day) by gavage for 24 weeks. The DM rats significantly increased the serum fasting blood glucose and lipid levels, which were alleviated by melatonin treatment. Importantly, the intestinal epithelial permeability was significantly increased in DM rats but was ameliorated following treatment with melatonin. These findings also indicated the expression of myosin light chain kinase (MLCK) and phosphorylation of MLC targeting subunit (MYPT) induced myosin light chain (MLC) phosphorylation level was markedly elevated in hyperglycemic and hyperlipidemic status. They were partly associated with down-regulated membrane type 1 and 2 (MT1 and MT2) expression, and up-regulated Rho-associated protein kinase (ROCK) expression and increased extracellular signal-regulated kinase (ERK) phosphorylation. However, the changes in target protein expression were reversed by melatonin. In conclusion, our results show melatonin beneficial effects on impaired intestinal epithelial permeability in T2D by suppressing ERK/MLCK- and ROCK/MCLP-dependent MLC phosphorylation.

  5. Heterologous expression of MlcE in Saccharomyces cerevisiae provides resistance to natural and semi-synthetic statins

    Directory of Open Access Journals (Sweden)

    Ana Ley

    2015-12-01

    Full Text Available Statins are inhibitors of 3-hydroxy-3-methylglutaryl coenzyme A reductase, the key enzyme in cholesterol biosynthesis. Their extensive use in treatment and prevention of cardiovascular diseases placed statins among the best selling drugs. Construction of Saccharomyces cerevisiae cell factory for the production of high concentrations of natural statins will require establishment of a non-destructive self-resistance mechanism to overcome the undesirable growth inhibition effects of statins. To establish active export of statins from yeast, and thereby detoxification, we integrated a putative efflux pump-encoding gene mlcE from the mevastatin-producing Penicillium citrinum into the S. cerevisiae genome. The resulting strain showed increased resistance to both natural statins (mevastatin and lovastatin and semi-synthetic statin (simvastatin when compared to the wild type strain. Expression of RFP-tagged mlcE showed that MlcE is localized to the yeast plasma and vacuolar membranes. We provide a possible engineering strategy for improvement of future yeast based production of natural and semi-synthetic statins.

  6. Immunomodulatory effects of oral antidiabetic drugs in lymphocyte cultures from patients with type 2 diabetes Efeito imunomodulador de hipoglicemiantes orais em cultura de linfócitos de pacientes com diabetes tipo 2

    OpenAIRE

    2011-01-01

    INTRODUCTION AND OBJECTIVE: It has been suggested that type 2 diabetes is an inflammatory response manifestation. The main drugs used to treat type 2 diabetes are sulphonylureas and biguanides. The aim of this study was to demonstrate the modulatory effects of oral hypoglycemic drugs (chlorpropamide and metformin) on lymphocyte proliferation in vitro and ex vivo. METHODS: Peripheral blood mononuclear cells were isolated from human blood by gradient centrifugation. T-lymphocytes were stimulate...

  7. Monte Carlo simulation of MLC-shaped TrueBeam electron fields benchmarked against measurement

    CERN Document Server

    Lloyd, Samantha AM; Zavgorodni, Sergei

    2014-01-01

    Modulated electron radiotherapy (MERT) and combined, modulated photon/electron radiotherapy (MPERT) have received increased research attention, having shown capacity for reduced low dose exposure to healthy tissue and comparable, if not improved, target coverage for a number of treatment sites. Accurate dose calculation tools are necessary for clinical treatment planning, and Monte Carlo (MC) is the gold standard for electron field simulation. With many clinics replacing older accelerators, MC source models of the new machines are needed for continued development, however, Varian has kept internal schematics of the TrueBeam confidential and electron phase-space sources have not been made available. TrueBeam electron fields are not substantially different from those generated by the Clinac 21EX, so we have modified the internal schematics of the Clinac 21EX to simulate TrueBeam electrons. BEAMnrc/DOSXYZnrc were used to simulate 5x5 and 20x20 cm$^2$ electron fields with MLC-shaped apertures. Secondary collimati...

  8. Schemes of Repeater Optimizing Distribution based on the MLC Application and CBLRD Simulation

    Directory of Open Access Journals (Sweden)

    Qian Qiuye

    2013-07-01

    Full Text Available The widely use of repeaters raises concern about their coordination among the public. Since repeaters may suffer interaction and limitation bearing capacity, designing a reasonable repeaters coordination method is of great significance. This study address the problem if repeater coordination in a circular flat area with minimal number of repeaters with seamless coverage theory, system simulation method. With 1,000 users, this study model the coverage, getting the minimal number of repeaters of different coverage radius based on extensive used regular hexagon coverage theory. A numerical example was given in this case. When the number of users increases to 10,000, this study simulate to get the signal density across the area according to the consideration of repeaters and the different distribution of users, which are divided into uniform distribution, linear distribution, normal distribution and lognormal distribution. Then, Multi-Layer Coverage (MLC and Coverage by Link Rate Density (CBLRD are created as the distribution scheme on the area where repeat service demand is large. Moreover, for solution on the distribution of the repeaters with barriers, distribution schemes are given considering the transmission of VHF spectrums and the distribution of users around the barrier. Additionally, Spring Comfortable Degree (SCD is used for evaluation of the results and the developing tends are given to improve the model. Due to the reasonable assumption, the solution of repeater distribution is of pivotal reference value based on the reasonable results.

  9. LYMPHOCYTE APOPTOSIS IN PSORIASIS

    Directory of Open Access Journals (Sweden)

    О. M. Kapuler

    2006-01-01

    Full Text Available Abstract. Forty-two patients with progressive vulgar psoriasis (PASI = 19.7 ± 1.5 and 40 healthy volunteers were under investigation. Psoriatic patients were characterized by increased number of CD4+ CD95+ peripheral blood T lymphocytes, which correlates with clinical psoriatic score, and by increased levels of soluble Fas (sFas in serum, as compared to controls (resp., 1868.1 ± 186.8 pg/ml vs. 1281.4 ± 142.5 pg/ml, PLSD = 0.019. The levels of spontaneous lymphocyte apoptosis and anti-Fas (Mab-induced apoptosis in psoriatic patients did not differ from the controls. However, apoptosis induced by “oxidative stress” (50 M Н202, 4 hrs was depressed in the patients. Moreover, a simultaneous assessment of cell cycle structure (metachromatic staining with Acridine Orange, apoptosis and Fas receptor expression (AnnV-FITC/antiFas mAbs-PE staining following a short-term mitogenic stimulation (PHA-P, 5 µg/ml, 24 hrs were performed. We found no marked differences in mitogenic reactivity, activation-induced apoptosis, and activation-induced Fas receptor expression when studying lymphocytes from healthy donors and psoriatic patients. However, PHA-activated lymphocytes from psoriatic patients displayed a significantly decreased ratio of AnnV+CD95+ to the total AnnV+ subpopulation, thus suggesting a decreased role of Fas-dependent mechanisms of apoptosis during the cell activation. The data obtained confirm a view, that an abnormal lymphocyte “apoptotic reactivity”, which plays a crucial role in the mechanisms of autoimmunity, may also of importance in the pathogenesis of psoriasis.

  10. Experimental investigation of a moving averaging algorithm for motion perpendicular to the leaf travel direction in dynamic MLC target tracking

    Energy Technology Data Exchange (ETDEWEB)

    Yoon, Jai-Woong; Sawant, Amit; Suh, Yelin; Cho, Byung-Chul; Suh, Tae-Suk; Keall, Paul [Department of Biomedical Engineering, College of Medicine, Catholic University of Korea, Seoul, Korea 131-700 and Research Institute of Biomedical Engineering, Catholic University of Korea, Seoul, 131-700 (Korea, Republic of); Department of Radiation Oncology, Stanford University, Stanford, California 94305 (United States); Department of Radiation Oncology, Stanford University, Stanford, California 94305 (United States) and Department of Radiation Oncology, Asan Medical Center, Seoul, 138-736 (Korea, Republic of); Department of Biomedical Engineering, College of Medicine, Catholic University of Korea, Seoul, 131-700 and Research Institute of Biomedical Engineering, Catholic University of Korea, Seoul, 131-700 (Korea, Republic of); Department of Radiation Oncology, Stanford University, Stanford, California 94305 (United States) and Radiation Physics Laboratory, Sydney Medical School, University of Sydney, 2006 (Australia)

    2011-07-15

    Purpose: In dynamic multileaf collimator (MLC) motion tracking with complex intensity-modulated radiation therapy (IMRT) fields, target motion perpendicular to the MLC leaf travel direction can cause beam holds, which increase beam delivery time by up to a factor of 4. As a means to balance delivery efficiency and accuracy, a moving average algorithm was incorporated into a dynamic MLC motion tracking system (i.e., moving average tracking) to account for target motion perpendicular to the MLC leaf travel direction. The experimental investigation of the moving average algorithm compared with real-time tracking and no compensation beam delivery is described. Methods: The properties of the moving average algorithm were measured and compared with those of real-time tracking (dynamic MLC motion tracking accounting for both target motion parallel and perpendicular to the leaf travel direction) and no compensation beam delivery. The algorithm was investigated using a synthetic motion trace with a baseline drift and four patient-measured 3D tumor motion traces representing regular and irregular motions with varying baseline drifts. Each motion trace was reproduced by a moving platform. The delivery efficiency, geometric accuracy, and dosimetric accuracy were evaluated for conformal, step-and-shoot IMRT, and dynamic sliding window IMRT treatment plans using the synthetic and patient motion traces. The dosimetric accuracy was quantified via a {gamma}-test with a 3%/3 mm criterion. Results: The delivery efficiency ranged from 89 to 100% for moving average tracking, 26%-100% for real-time tracking, and 100% (by definition) for no compensation. The root-mean-square geometric error ranged from 3.2 to 4.0 mm for moving average tracking, 0.7-1.1 mm for real-time tracking, and 3.7-7.2 mm for no compensation. The percentage of dosimetric points failing the {gamma}-test ranged from 4 to 30% for moving average tracking, 0%-23% for real-time tracking, and 10%-47% for no compensation

  11. Influence of gamma-irradiation on telomere length of cultured human lymphocytes%γ射线辐射对人淋巴细胞端粒长度影响研究

    Institute of Scientific and Technical Information of China (English)

    郭月凤; 张睿凤; 张慧芳

    2015-01-01

    目的:探讨γ射线辐射对人淋巴细胞平均端粒长度的影响。方法培养非洲淋巴细胞瘤病毒转化正常人B淋巴细胞( Peng-EBV细胞),接受钴-60γ射线照射,剂量率为0.96 Gy/min,吸收剂量分别为0.2、0.6、1.6、3.0和6.0 Gy;以不进行照射的细胞作为对照组。分别收集照射后24、48和72 h细胞样品,提取基因组DNA。采用定量聚合酶链式反应( Q-PCR)法检测样品平均端粒长度。分析γ辐射影响人淋巴细胞端粒长度的剂量-效应和时间-效应。结果端粒标准DNA的标准曲线为Ct=B+M ×lg D=35.264-3.339 lg D(相关系数=0.964,P<0.05)。γ射线单次照射可导致Peng-EBV细胞在照射后24~72 h端粒长度较同时间点对照组延长(P<0.05)。在0.0~1.6 Gy剂量范围内,γ射线诱导的Peng-EBV细胞端粒长度在照射后48 h有随剂量增加而增加的趋势(P<0.05)。0.6和1.6 Gy 2个剂量组的端粒延长幅度有随照后时间延长而增大的趋势(P<0.05)。结论Q-PCR方法适用于淋巴细胞绝对端粒长度检测。γ射线单次照射可以导致Peng-EBV细胞平均端粒长度增加。%Objective To investigate the effect of 60 Co γ-ray irradiation on average telomere length of human lymphocytes.Methods Epstein-Barr virus transformated human B homeocyte lines ( Peng-EBV cells) were cultured, and then were exposed to 60 Coγ-ray at the dose rate of 0.96 Gy/min.The absorbed doses were 0.2, 0.6, 1.6, 3.0, and 6.0 Gy, respectively.The Peng-EBV cells without irradiation exposure were selected as control group.The sample cells were collected after irradiation for 24, 48, and 72 hours respectively and genome DNA was extracted.The average telomere length of samples was measured by quantitative polymerase chain reaction ( Q-PCR) .Then, the time-effect and dose-effect of 60 Co γ-ray irradiation on the telomere length of human lymphocytes

  12. Analysis of direct clinical consequences of MLC positional errors in volumetric-modulated arc therapy using 3D dosimetry system.

    Science.gov (United States)

    Nithiyanantham, Karthikeyan; Mani, Ganesh K; Subramani, Vikraman; Mueller, Lutz; Palaniappan, Karrthick K; Kataria, Tejinder

    2015-09-08

    In advanced, intensity-modulated external radiotherapy facility, the multileaf collimator has a decisive role in the beam modulation by creating multiple segments or dynamically varying field shapes to deliver a uniform dose distribution to the target with maximum sparing of normal tissues. The position of each MLC leaf has become more critical for intensity-modulated delivery (step-and-shoot IMRT, dynamic IMRT, and VMAT) compared to 3D CRT, where it defines only field boundaries. We analyzed the impact of the MLC positional errors on the dose distribution for volumetric-modulated arc therapy, using a 3D dosimetry system. A total of 15 VMAT cases, five each for brain, head and neck, and prostate cases, were retrospectively selected for the study. All the plans were generated in Monaco 3.0.0v TPS (Elekta Corporation, Atlanta, GA) and delivered using Elekta Synergy linear accelerator. Systematic errors of +1, +0.5, +0.3, 0, -1, -0.5, -0.3 mm were introduced in the MLC bank of the linear accelerator and the impact on the dose distribution of VMAT delivery was measured using the COMPASS 3D dosim-etry system. All the plans were created using single modulated arcs and the dose calculation was performed using a Monte Carlo algorithm in a grid size of 3 mm. The clinical endpoints D95%, D50%, D2%, and Dmax,D20%, D50% were taken for the evaluation of the target and critical organs doses, respectively. A significant dosimetric effect was found for many cases even with 0.5 mm of MLC positional errors. The average change of dose D 95% to PTV for ± 1 mm, ± 0.5 mm, and ±0.3mm was 5.15%, 2.58%, and 0.96% for brain cases; 7.19%, 3.67%, and 1.56% for head and neck cases; and 8.39%, 4.5%, and 1.86% for prostate cases, respectively. The average deviation of dose Dmax was 5.4%, 2.8%, and 0.83% for brainstem in brain cases; 8.2%, 4.4%, and 1.9% for spinal cord in H&N; and 10.8%, 6.2%, and 2.1% for rectum in prostate cases, respectively. The average changes in dose followed a linear

  13. Comparison of conventional inserts and an add-on electron MLC for chest wall irradiation of left-sided breast cancer

    Energy Technology Data Exchange (ETDEWEB)

    Vatanen, Tero; Lahtinen, Tapani (Dept. of Oncology, Kuopio Univ. Hospital, Kuopio (Finland)); Traneus, Erik (Nucletron Scandinavia AB, Uppsala (Sweden))

    2009-04-15

    Background. Collimation of irregularly shaped clinical electron beams is currently based on electron inserts made of low melting point alloys. The present investigation compares a conventional electron applicator with insert and add-on eMLC-based dose distributions in the postoperative chest wall irradiation of left-sided breast cancer. Material and methods. Voxel Monte Carlo++ (VMC++) calculated dose distributions related to electron fields were compared with 10 left-sided breast cancer patients after radical mastectomy. The prescription dose was 50 Gy at a build-up maximum. The same dose was prescribed for the ipsilateral axillary, parasternal and supraclavicular lymph nodes that were treated with photons and calculated with a pencil beam algorithm. The insert beams were shaped with 1.5 cm thick Wood's metal electron inserts in an electron applicator of a Varian 2100 C/D linac. Doses for the eMLC-shaped beams were calculated for an eMLC prototype with 2 cm thick and 5 mm wide steel leaves. The same collimator-to-surface distance (CSD) of 5.8 cm was used for both collimators. Results. The mean PTV dose was slightly higher for the eMLC plans (50.7 vs 49.5 Gy, p<0.001, respectively). The maximum doses assessed by D5% for the eMLC and insert were 60.9 and 59.1 Gy (p<0.001). The difference was due to the slightly higher doses near the field edges for the eMLC. The left lung V20 volumes were 34.5% and 34.0% (p<0.001). There was only a marginal difference in heart doses. Discussion: Despite a slight increase of maximum dose in PTV the add-on electron MLC for chest wall irradiation results in practically no differences in dose distributions compared with the present insert-based collimation.

  14. cultural

    Directory of Open Access Journals (Sweden)

    Irene Kreutz

    2006-01-01

    Full Text Available Es un estudio cualitativo que adoptó como referencial teorico-motodológico la antropología y la etnografía. Presenta las experiencias vivenciadas por mujeres de una comunidad en el proceso salud-enfermedad, con el objetivo de comprender los determinantes sócio-culturales e históricos de las prácticas de prevención y tratamiento adoptados por el grupo cultural por medio de la entrevista semi-estructurada. Los temas que emergieron fueron: la relación entre la alimentación y lo proceso salud-enfermedad, las relaciones con el sistema de salud oficial y el proceso salud-enfermedad y lo sobrenatural. Los dados revelaron que los moradores de la comunidad investigada tienen un modo particular de explicar sus procedimientos terapéuticos. Consideramos que es papel de los profesionales de la salud en sus prácticas, la adopción de abordajes o enfoques que consideren al individuo en su dimensión sócio-cultural e histórica, considerando la enorme diversidad cultural en nuestro país.

  15. Serial tomotherapy vs. MLC-IMRT (Multileaf Collimator Intensity Modulated Radiotherapy) for simultaneous boost treatment large intracerebral lesions; Serielle Tomotherapie vs. MLC-IMRT (Multileaf Collimator Intensity Modulated Radiotherapy) fuer die simultane Boostbestrahlung mehrerer groesserer Hirnfiliae

    Energy Technology Data Exchange (ETDEWEB)

    Wolff, Dirk; Lohr, Frank; Mai, Sabine; Polednik, Martin; Wenz, Frederik [Universitaetsklinikum Mannheim, Mannheim (Germany). Klinik fuer Strahlentherapie und Radioonkologie; Abo-Madyan, Yasser [Universitaetsklinikum Mannheim, Mannheim (Germany). Klinik fuer Strahlentherapie und Radioonkologie; Kasr-El-Einy Hospital. Cairo Univ. (EG). Dept. of Oncology (NEMROCK); Dobler, Barbara [Klinikum der Universitaet Regensburg (Germany). Strahlentherapie

    2009-07-01

    Introduction: Recent data suggest that a radiosurgery boost treatment for up to three brain metastases in addition to whole brain radiotherapy (WBRT) is beneficial. Sequential treatment of multiple metastatic lesions is time-consuming and optimal normal tissue sparing is not trivial for larger metastases when separate plans are created and are only superimposed afterwards. Sequential Tomotherapy with noncoplanar arcs and Multi-field IMRT may streamline the process and enable easy simultaneous treatment. We compared plans for 2-3 intracerebral targets calculated with Intensity Modulated Radiotherapy (IMRT) based on treatment with MLC or sequential Tomotherapy using the Peacock-System. Treatment time was not to exceed 90 min on a linac with standart dose rate. MIMiC plans without treatment-time restrictions were created as a benchmark. Materials and methods: Calculations are based on a Siemens KD2 linac with a dose rate of 200 MU/min. Step-and-Shoot IMRT is performed with a standard MLC (2 x 29 leaves, 1 cm), serial Tomotherapy with the Multivane-Collimator MIMiC (NOMOS Inc. USA). Treatment plans are created with Corvus 5.0. To create plans with good conformity we chose a noncoplanar beam- and arc geometry for each approach (IMRT 4-, MIMiC 5-couch angles). The benchmark MIMiC plans with maximally steep dose gradients had 9 couch angles. For plan comparison reasons, 10Gy were prescribed to 90% of the PTV. Steepness of dose gradients, homogeneity and conformity were assessed by the following parameters: Volume encompassed by certain isodoses outside the target as well as homogeneity and conformity as indicated by Homogeneity- and Conformity-Index. Results: Plans without treatment-time restrictions had slightest dose to organ at risk (OAR), normal tissue and least Conformity-index. MIMiC- and MLC-IMRT based plans can be treated within the intended period of 90 min, all plans met the required dose. MLC based plans resulted in higher dose to organs at risk (OAR) and dose

  16. Implications of a high-definition multileaf collimator (HD-MLC on treatment planning techniques for stereotactic body radiation therapy (SBRT: a planning study

    Directory of Open Access Journals (Sweden)

    Chen Yiyi

    2009-07-01

    Full Text Available Abstract Purpose To assess the impact of two multileaf collimator (MLC systems (2.5 and 5 mm leaf widths on three-dimensional conformal radiotherapy, intensity-modulated radiotherapy, and dynamic conformal arc techniques for stereotactic body radiation therapy (SBRT of liver and lung lesions. Methods Twenty-nine SBRT plans of primary liver (n = 11 and lung (n = 18 tumors were the basis of this study. Five-millimeter leaf width 120-leaf Varian Millennium (M120 MLC-based plans served as reference, and were designed using static conformal beams (3DCRT, sliding-window intensity-modulated beams (IMRT, or dynamic conformal arcs (DCA. Reference plans were either re-optimized or recomputed, with identical planning parameters, for a 2.5-mm width 120-leaf BrainLAB/Varian high-definition (HD120 MLC system. Dose computation was based on the anisotropic analytical algorithm (AAA, Varian Medical Systems with tissue heterogeneity taken into account. Each plan was normalized such that 100% of the prescription dose covered 95% of the planning target volume (PTV. Isodose distributions and dose-volume histograms (DVHs were computed and plans were evaluated with respect to target coverage criteria, normal tissue sparing criteria, as well as treatment efficiency. Results Dosimetric differences achieved using M120 and the HD120 MLC planning were generally small. Dose conformality improved in 51.7%, 62.1% and 55.2% of the IMRT, 3DCRT and DCA cases, respectively, with use of the HD120 MLC system. Dose heterogeneity increased in 75.9%, 51.7%, and 55.2% of the IMRT, 3DCRT and DCA cases, respectively, with use of the HD120 MLC system. DVH curves demonstrated a decreased volume of normal tissue irradiated to the lower (90%, 50% and 25% isodose levels with the HD120 MLC. Conclusion Data derived from the present comparative assessment suggest dosimetric merit of the high definition MLC system over the millennium MLC system. However, the clinical significance of these results

  17. SU-E-T-175: Evaluation of the Relative Output Ratio for Collimator Jaw and MLC Defined Small Static 6MV Photon Fields

    Energy Technology Data Exchange (ETDEWEB)

    Cho, G [Chris O' Brien Lifehouse at RPAH, Camperdown, NSW 2050 (Australia); Institute of Medical Physics, School of Physics, University of Sydney, Sydney, NSW (Australia); Thwaites, D [Institute of Medical Physics, School of Physics, University of Sydney, Sydney, NSW (Australia)

    2014-06-01

    Purpose: To evaluate relative output ratio of collimator jaw and MLC defined small photon fields. Methods: Relative output ratios were measured using Gafchromic EBT3 film for a 6 MV photon beam on a Novalis Tx with HD120 MLC. Beam collimation was achieved by the jaws for 1.0 cm and 3.0 cm and MLC defined square field sizes between 0.5 cm and 1.0 cm with varying jaw settings between 2.0 and 4.0 cm. Film pieces were exposed to 4 Gy. Experiments were repeated with each session consisting of five consecutive exposures for the given MLC and/or jaw collimation and with the MLC and the jaws reset for each exposure. Films were scanned using EPSON 10000XL flatbed scanner approximately 24 hours after exposure in 48 bit RGB format at 150 dpi. Film calibration data were corrected for daily linac output variations. Doses were evaluated using the green channel with square ROI sizes of 0.1 – 0.6 cm. Converted doses were normalised for output ratio calculation using the 3.0 cm field as a machine specific reference field size. Mean output ratio and coefficient of variation (CV) were calculated for each experimental session. Results: For the Novalis 6 MV photon beam the output ratios between 0.719 and 0.872 have been measured for the jaw/MLC combinations tested. For a jaw setting of 4.0 cm field, the mean CV of the output ratios increased from 0.77% to 1.48% with decreasing MLC field size from 1.0 cm to 0.5 cm. For a nominal MLC 1.0 cm field, the CV increased to 1.00% from 0.77% with reducing jaw field size from 4.0 cm to 2.0 cm. Conclusion: The relative output ratio and the associated CV were dependent on the collimator jaw and MLC settings. The field size dependent CV showed similar trends to those reported in the literature.

  18. Evaluation of the dose calculation accuracy for small fields defined by jaw or MLC for AAA and Acuros XB algorithms.

    Science.gov (United States)

    Fogliata, Antonella; Lobefalo, Francesca; Reggiori, Giacomo; Stravato, Antonella; Tomatis, Stefano; Scorsetti, Marta; Cozzi, Luca

    2016-10-01

    Small field measurements are challenging, due to the physical characteristics coming from the lack of charged particle equilibrium, the partial occlusion of the finite radiation source, and to the detector response. These characteristics can be modeled in the dose calculations in the treatment planning systems. Aim of the present work is to evaluate the MU calculation accuracy for small fields, defined by jaw or MLC, for anisotropic analytical algorithm (AAA) and Acuros XB algorithms, relative to output measurements on the beam central axis. Single point output factor measurement was acquired with a PTW microDiamond detector for 6 MV, 6 and 10 MV unflattened beams generated by a Varian TrueBeam STx equipped with high definition-MLC. Fields defined by jaw or MLC apertures were set; jaw-defined: 0.6 × 0.6, 0.8 × 0.8, 1 × 1, 2 × 2, 3 × 3, 4 × 4, 5 × 5, and 10 × 10 cm(2); MLC-defined: 0.5 × 0.5 cm(2) to the maximum field defined by the jaw, with 0.5 cm stepping, and jaws set to: 2 × 2, 3 × 3, 4 × 4, 5 × 5, and 10 × 10 cm(2). MU calculation was obtained with 1 mm grid in a virtual water phantom for the same fields, for AAA and Acuros algorithms implemented in the Varian eclipse treatment planning system (version 13.6). Configuration parameters as the effective spot size (ESS) and the dosimetric leaf gap (DLG) were varied to find the best parameter setting. Differences between calculated and measured doses were analyzed. Agreement better than 0.5% was found for field sizes equal to or larger than 2 × 2 cm(2) for both algorithms. A dose overestimation was present for smaller jaw-defined fields, with the best agreement, averaged over all the energies, of 1.6% and 4.6% for a 1 × 1 cm(2) field calculated by AAA and Acuros, respectively, for a configuration with ESS = 1 mm for both X and Y directions for AAA, and ESS = 1.5 and 0 mm for X and Y directions for Acuros. Conversely, a calculated dose underestimation was found for small MLC-defined fields, with the

  19. Benzene metabolites induce apoptosis in lymphocytes.

    Science.gov (United States)

    Martínez-Velázquez, M; Maldonado, V; Ortega, A; Meléndez-Zajgla, J; Albores, A

    2006-08-01

    Benzene is an important environmental pollutant with important health implications. Exposure to this aromatic hydrocarbon is associated with hematotoxicity, and bone marrow carcinogenic effects. It has been shown that benzene induces oxidative stress, cell cycle alterations, and programmed cell death in cultured cells. Hepatic metabolism of benzene is thought to be a prerequisite for its bone marrow toxicity. Nevertheless, there are no reports on the cellular effects of reactive intermediates derived from hepatic metabolism of benzene. Thus, the goal of this project was to determine the cellular alterations of benzene metabolites produced by the cultured hepatic cell line HepG2. Supernatants collected from these cells were applied to a culture of freshly isolated lymphocytes. A higher decrease in cell viability was found in cells exposed to these supernatants than to unmetabolized benzene. This viability decrease was due to apoptosis, as determined by Terminal deoxynucleotidyl Transferase Biotin-dUTP Nick End Labeling (TUNEL) assay and internucleosomal fragmentation of DNA. When supernatants were analyzed by HPLC, we found that not all the hydrocarbon was biotransformed, since a 28 microM concentration (37%) remained. The only metabolite found in the culture medium was muconic acid. The present results show that muconic acid derived from benzene metabolism is able to cooperate with the pollutant for the induction of apoptosis in rat lymphocytes.

  20. Effects of low dose oxymatrine on mouse lymphocyte proliferation stimulated by Con A

    Institute of Scientific and Technical Information of China (English)

    LI Luo-si; WU Bin; LI Jian-guo; XIE Hong-fu; ZHANG Yang-de; CONG Ling; SHI Jun

    2005-01-01

    Objective To investigate the effects of low dose Oxymatrine (OMT) on mouse lymphocyte proliferation stimulated by Con A making use of fluorescence dyestuff CFDA-SE. Methods CFDA-SE staining and flow cytometry were used to detect the fluorescence intensity of lymphocytes after stimulated by polyclonal stimulators Con A and OMT. Then, related software was used to analyze the effects of OMT on mouse lymphocyte proliferation.Results After cultured for 48 h, CFSE fluorescence could be detected by cytometer, filial generation peaks did not appear in control group, which indicated that lymphocytes did not proliferate. Three peaks were obviously detected in Con A group which indicated that Lymphocytes divided after 48 h stimulated by Con A compared with the halving of the fluorescence intensity of control group. In groups with Con A and OMT treated, Primary generation peaks are all lower while filial generation peaks are significantly higher than groups with Con A treated only. This indicated OMT obviously promote lymphocyte proliferation. After cultured for 72 h, the fluorescence intensity changes between all groups are consistent with those of cultured for 48 h. Analyzed with CELLQuest software, it is shown that OMT could promote lymphocyte proliferation in 16, 8, 4 and 2μg/mL respectively. Conclusions 1) CFDA-SEdyeing and flow cytometer were both reliable tools to analyze lymphocyte proliferation; 2) lower dosage of OMTcould promote the proliferation of lymphocyte as a immunopotentiator.

  1. Curcumin and Cholecalciferol in Treating Patients With Previously Untreated Stage 0-II Chronic Lymphocytic Leukemia or Small Lymphocytic Lymphoma

    Science.gov (United States)

    2016-10-04

    Contiguous Stage II Small Lymphocytic Lymphoma; Noncontiguous Stage II Small Lymphocytic Lymphoma; Stage 0 Chronic Lymphocytic Leukemia; Stage I Chronic Lymphocytic Leukemia; Stage I Small Lymphocytic Lymphoma; Stage II Chronic Lymphocytic Leukemia

  2. Dynamic MLC tracking of moving targets with a single kV imager for 3D conformal and IMRT treatments

    Energy Technology Data Exchange (ETDEWEB)

    Poulsen, Per R. (Dept. of Oncology, Aarhus Univ. Hospital (Denmark)), E-mail: perpolse@rm.dk; Cho, Byungchul; Sawant, Amit; Ruan, Dan; Keall, Paul J. (Dept. of Radiation Oncology, Stanford Univ., Stanford (United States))

    2010-10-15

    Background. Tumor motion during radiotherapy is a major challenge for accurate dose delivery, in particular for hypofractionation and dose painting. The motion may be compensated by dynamic multileaf collimator (DMLC) tracking. Previous work has demonstrated that a single kV imager can accurately localize moving targets for DMLC tracking during rotational delivery, however this method has not been investigated for the static gantry geometry used for conformal and IMRT treatments. In this study we investigate the accuracy of single kV-imager based DMLC tracking for static-gantry delivery. Material and methods. A 5-field treatment plan with circular field shape and 200 MU per field was delivered in 20 s per field to a moving phantom with an embedded gold marker. Fluoroscopic kV images were acquired at 5 Hz perpendicular to the treatment beam axis during a 120 deg pre-treatment gantry rotation, during treatment delivery, and during inter-field gantry rotations. The three-dimensional marker position was estimated from the kV images and used for MLC adaptation. Experiments included 12 thoracic/abdominal tumor trajectories and five prostate trajectories selected from databases with 160 and 548 trajectories, respectively. The tracking error was determined as the mismatch between the marker position and the MLC aperture center in portal images. Simulations extended the study to all trajectories in the databases and to treatments with prolonged duration of 60 s per field. Results. In the experiments, the mean root-mean-square (rms) tracking error was 0.9 mm (perpendicular to MLC) and 1.1 mm (parallel to MLC) for thoracic/abdominal tumor trajectories and 0.6 mm (perpendicular) and 0.5 mm (parallel) for prostate trajectories. Simulations of these experiments agreed to within 0.1 mm. Simulations of all trajectories in the databases resulted in mean rms tracking errors of 0.6 mm (perpendicular) and 0.9 mm (parallel) for thorax/abdomen tumors and 0.4 mm (perpendicular) and 0

  3. [The effect of mixed cultivation of lymphocytes irradiated at a dose of 1 Gy and non-irradiated lymphocytes on the frequency of chromosomal aberration].

    Science.gov (United States)

    Kolesnikova, I S

    2012-01-01

    The mutual influence of irradiated (1 Gy) and non-irradiated cells was demonstrated on the model of the mixed culture oflymphocytes from opposite gender donors using chromosome aberrations (ChA) as an endpoint. The number of ChA in non-irradiated lymphocytes in mixed cultures with irradiated ones increased as compared to the corresponding monocultures. At the same time, the number of induced ChA decreased in the irradiated lymphocytes cultivated with non-irradiated ones.

  4. Lymphocyte 'homing' and chronic inflammation.

    Science.gov (United States)

    Sakai, Yasuhiro; Kobayashi, Motohiro

    2015-07-01

    Chronic inflammation is a response to prolonged exposure to injurious stimuli that harm and destroy tissues and promote lymphocyte infiltration into inflamed sites. Following progressive accumulation of lymphocytes, the histology of inflamed tissue begins to resemble that of peripheral lymphoid organs, which can be referred to as lymphoid neogenesis or formation of tertiary lymphoid tissues. Lymphocyte recruitment to inflamed tissues is also reminiscent of lymphocyte homing to peripheral lymphoid organs. In the latter, under physiological conditions, homing receptors expressed on lymphocytes adhere to vascular addressin expressed on high endothelial venules (HEVs), initiating a lymphocyte migration process composed of sequential adhesive interactions. Intriguingly, in chronic inflammation, HEV-like vessels are induced de novo, despite the fact that the inflamed site is not originally lymphoid tissue, and these vessels contribute to lymphocyte recruitment in a manner similar to physiological lymphocyte homing. In this review, we first describe physiological lymphocyte homing mechanisms focusing on vascular addressins. We then describe HEV-like vessel-mediated pathogenesis seen in various chronic inflammatory disorders such as Helicobacter pylori gastritis, inflammatory bowel disease (IBD), autoimmune pancreatitis and sclerosing sialadenitis, as well as chronic inflammatory cell neoplasm MALT lymphoma, with reference to our work and that of others.

  5. Lymphocyte Trafficking to Mucosal Tissues

    DEFF Research Database (Denmark)

    Mikhak, Zamaneh; Agace, William Winston; Luster, Andrew D.

    2015-01-01

    Lymphocytes are the key cells of the adaptive immune system that provide antigen-specific responses tailored to the context of antigen exposure. Through cytokine release and antibody production, lymphocytes orchestrate and amplify the recruitment and function of other immune cells and contribute...... to host defense against invading pathogens and the pathogenesis of many inflammatory diseases. Lymphocyte function is critically dependent on their ability to traffic into the correct anatomic locations at the appropriate times. This process is highly regulated and requires that lymphocytes interact...

  6. Lymphocytic Interstitial Pneumonia.

    Science.gov (United States)

    Panchabhai, Tanmay S; Farver, Carol; Highland, Kristin B

    2016-09-01

    Lymphocytic interstitial pneumonia (LIP) is a rare lung disease on the spectrum of benign pulmonary lymphoproliferative disorders. LIP is frequently associated with connective tissue diseases or infections. Idiopathic LIP is rare; every attempt must be made to diagnose underlying conditions when LIP is diagnosed. Computed tomography of the chest in patients with LIP may reveal ground-glass opacities, centrilobular and subpleural nodules, and randomly distributed thin-walled cysts. Demonstrating polyclonality with immunohistochemistry is the key to differentiating LIP from lymphoma. The 5-year mortality remains between 33% and 50% and is likely to vary based on the underlying disease process.

  7. Chronic lymphocytic leukaemia

    Science.gov (United States)

    Kipps, Thomas J.; Stevenson, Freda K.; Wu, Catherine J.; Croce, Carlo M.; Packham, Graham; Wierda, William G.; O’Brien, Susan; Gribben, John; Rai, Kanti

    2017-01-01

    Chronic lymphocytic leukaemia (CLL) is a malignancy of CD5+ B cells that is characterized by the accumulation of small, mature-appearing lymphocytes in the blood, marrow and lymphoid tissues. Signalling via surface immunoglobulin, which constitutes the major part of the B cell receptor, and several genetic alterations play a part in CLL pathogenesis, in addition to interactions between CLL cells and other cell types, such as stromal cells, T cells and nurse-like cells in the lymph nodes. The clinical progression of CLL is heterogeneous and ranges from patients who require treatment soon after diagnosis to others who do not require therapy for many years, if at all. Several factors, including the immunoglobulin heavy-chain variable region gene (IGHV) mutational status, genomic changes, patient age and the presence of comorbidities, should be considered when defining the optimal management strategies, which include chemotherapy, chemoimmunotherapy and/or drugs targeting B cell receptor signalling or inhibitors of apoptosis, such as BCL-2. Research on the biology of CLL has profoundly enhanced our ability to identify patients who are at higher risk for disease progression and our capacity to treat patients with drugs that selectively target distinctive phenotypic or physiological features of CLL. How these and other advances have shaped our current understanding and treatment of patients with CLL is the subject of this Primer. PMID:28102226

  8. Decreased deformability of lymphocytes in chronic lymphocytic leukemia

    Science.gov (United States)

    Zheng, Yi; Wen, Jun; Nguyen, John; Cachia, Mark A.; Wang, Chen; Sun, Yu

    2015-01-01

    This paper reports the first study of stiffness/deformability changes of lymphocytes in chronic lymphocytic leukemia (CLL) patients, demonstrating that at the single cell level, leukemic metastasis progresses are accompanied by biophysical property alterations. A microfluidic device was utilized to electrically measure cell volume and transit time of single lymphocytes from healthy and CLL patients. The results from testing thousands of cells reveal that lymphocytes from CLL patients have higher stiffness (i.e., lower deformability), as compared to lymphocytes in healthy samples, which was also confirmed by AFM indentation tests. This observation is in sharp contrast to the known knowledge on other types of metastatic cells (e.g., breast and lung cancer cells) whose stiffness becomes lower as metastasis progresses.

  9. Activation of lymphocytes induced by bronchial epithelial cells with prolonged RSV infection.

    Directory of Open Access Journals (Sweden)

    Ling Qin

    Full Text Available Respiratory syncytial virus (RSV preferentially infects airway epithelial cells,which might be responsible for susceptibility to asthma; however, the underlying mechanism is not clear. This study determined the activation of lymphocytes and drift of helper T (Th subsets induced by RSV-infected human bronchial epithelial cells (HBECs in vitro. HBECs had prolonged infection with RSV, and lymphocytes isolated from human peripheral blood were co-cultured with RSV-infected HBECs. Four groups were established, as follows: lymphocytes (group L; lymphocytes infected with RSV (group RL; co-culture of lymphocytes with non-infected HBECs (group HL; and co-culture of lymphocytes with infected HBECs (group HRL. After co-culture with HBECs for 24 hours, lymphocytes were collected and the following were determined in the 4 groups: cell cycle status; apoptosis rate; and concentrations of IL-4, IFN-γ, and IL-17 in the supernatants. Cell cycle analysis for lymphocytes showed a significant increase in S phase cells, a decrease in G1 phase cells, and a higher apoptosis rate in group HRL compared with the other three groups. In group HRL, the levels of IL-4, IFN-γ, and IL-17 in supernatants were also higher than the other three groups. For further study, lymphocytes were individually treated with supernatants from non-infected and RSV-infected HBECs for 24 h. We showed that supernatants from RSV-infected HBECs induced the differentiation of Th2 and Th17 subsets, and suppressed the differentiation of Treg subsets. Our results showed that HBECs with prolonged RSV infection can induce lymphocyte proliferation and apoptosis, and enhance the release of cytokines by lymphocytes. Moreover, subset drift might be caused by RSV-infected HBECs.

  10. Asymmetric Programming: A Highly Reliable Metadata Allocation Strategy for MLC NAND Flash Memory-Based Sensor Systems

    Science.gov (United States)

    Huang, Min; Liu, Zhaoqing; Qiao, Liyan

    2014-01-01

    While the NAND flash memory is widely used as the storage medium in modern sensor systems, the aggressive shrinking of process geometry and an increase in the number of bits stored in each memory cell will inevitably degrade the reliability of NAND flash memory. In particular, it's critical to enhance metadata reliability, which occupies only a small portion of the storage space, but maintains the critical information of the file system and the address translations of the storage system. Metadata damage will cause the system to crash or a large amount of data to be lost. This paper presents Asymmetric Programming, a highly reliable metadata allocation strategy for MLC NAND flash memory storage systems. Our technique exploits for the first time the property of the multi-page architecture of MLC NAND flash memory to improve the reliability of metadata. The basic idea is to keep metadata in most significant bit (MSB) pages which are more reliable than least significant bit (LSB) pages. Thus, we can achieve relatively low bit error rates for metadata. Based on this idea, we propose two strategies to optimize address mapping and garbage collection. We have implemented Asymmetric Programming on a real hardware platform. The experimental results show that Asymmetric Programming can achieve a reduction in the number of page errors of up to 99.05% with the baseline error correction scheme. PMID:25310473

  11. The effect of extremely narrow MLC leaf width on the plan quality of VMAT for prostate cancer.

    Science.gov (United States)

    Park, Jong Min; Park, So-Yeon; Kim, Jin Ho; Carlson, Joel; Kim, Jung-In

    2016-06-23

    To investigate the effect of multi-leaf collimators (MLCs) with leaf width of 1.25 mm on the plan quality of volumetric modulated arc therapy (VMAT) for prostate cancer. A total of 20 patients with prostate cancer were retrospectively selected. Using a high definition MLC (HD MLC), primary and boost VMAT plans with two full arcs were generated for each patient (original plan). After that, by shifting the isocenter position of the 2nd arc by 1.25 mm in the cranio-caudal direction, we simulated fluences made with MLCs with leaf width of 1.25 mm. After shifting, primary and boost plans were generated for each patient (shifted plan). A sum plan was generated by summation of the primary and boost plan for each patient. Dose-volumetric parameters were calculated and compared. Both the homogeneity index (HI) and conformity index (CI) of the shifted plans were better than those of the original plans in primary plans (HI = 0.065 vs. 0.059 with p plans for the boost target volume showed better homogeneity and conformity than did the original plans (HI = 0.060 vs. 0.053 with p plans was closer to the prescription dose than that of the shifted plans in the case of sum plans (81.45 Gy vs. 81.12 Gy with p = 0.001). Use of extremely narrow MLCs could increase dose homogeneity and conformity of the target volume for prostate VMAT.

  12. Honokiol inhibits migration of renal cell carcinoma through activation of RhoA/ROCK/MLC signaling pathway.

    Science.gov (United States)

    Cheng, Shujie; Castillo, Victor; Welty, Matt; Eliaz, Isaac; Sliva, Daniel

    2016-10-01

    Honokiol, a biologically active compound isolated from Magnolia bark, has been shown to possess promising anticancer effect through induction of apoptosis. However, there is a relative lack of information regarding its anti‑metastatic activity. Renal cell carcinoma (RCC) is the most common malignancy of the adult kidney and is known for high risk of metastasis. Clinically, therapeutic methods for metastatic RCC cases are limited and efforts to exploit new treatments are still ongoing. The results of our current investigation first revealed that honokiol suppressed the proliferation of different human RCCs without affecting cell viability. In addition, honokiol inhibited migration of highly metastatic RCC 786‑0 cells and stimulated the activity of small GTPase, RhoA. Furthermore, phosphorylated myosin light chain (MLC) and excessive formation of actin stress fibers were identified in 786‑0 cells treated with honokiol. Interestingly, the pharmacological Rho‑associated protein kinase (ROCK) inhibitor Y‑27632 attenuated contraction of actin stress fibers induced by honokiol and abrogated honokiol‑mediated inhibition of cell migration. Together these important findings suggest that honokiol suppresses the migration of highly metastatic RCC through activation of RhoA/ROCK/MLC signaling and warrants attention in the treatment of RCC metastasis as a novel therapeutic approach.

  13. Dose-to-water conversion for the backscatter-shielded EPID: A frame-based method to correct for EPID energy response to MLC transmitted radiation

    Energy Technology Data Exchange (ETDEWEB)

    Zwan, Benjamin J., E-mail: benjamin.zwan@uon.edu.au; O’Connor, Daryl J. [School of Mathematical and Physical Sciences, University of Newcastle, Newcastle, New South Wales 2308 (Australia); King, Brian W. [Department of Radiation Oncology, Massey Cancer Center, Virginia Commonwealth University Health System, Richmond, Virginia 23298 (United States); Greer, Peter B. [School of Mathematical and Physical Sciences, University of Newcastle, Newcastle, New South Wales 2308, Australia and Department of Radiation Oncology, Calvary Mater Newcastle Hospital, Newcastle, New South Wales 2310 (Australia)

    2014-08-15

    Purpose: To develop a frame-by-frame correction for the energy response of amorphous silicon electronic portal imaging devices (a-Si EPIDs) to radiation that has transmitted through the multileaf collimator (MLC) and to integrate this correction into the backscatter shielded EPID (BSS-EPID) dose-to-water conversion model. Methods: Individual EPID frames were acquired using a Varian frame grabber and iTools acquisition software then processed using in-house software developed inMATLAB. For each EPID image frame, the region below the MLC leaves was identified and all pixels in this region were multiplied by a factor of 1.3 to correct for the under-response of the imager to MLC transmitted radiation. The corrected frames were then summed to form a corrected integrated EPID image. This correction was implemented as an initial step in the BSS-EPID dose-to-water conversion model which was then used to compute dose planes in a water phantom for 35 IMRT fields. The calculated dose planes, with and without the proposed MLC transmission correction, were compared to measurements in solid water using a two-dimensional diode array. Results: It was observed that the integration of the MLC transmission correction into the BSS-EPID dose model improved agreement between modeled and measured dose planes. In particular, the MLC correction produced higher pass rates for almost all Head and Neck fields tested, yielding an average pass rate of 99.8% for 2%/2 mm criteria. A two-sample independentt-test and fisher F-test were used to show that the MLC transmission correction resulted in a statistically significant reduction in the mean and the standard deviation of the gamma values, respectively, to give a more accurate and consistent dose-to-water conversion. Conclusions: The frame-by-frame MLC transmission response correction was shown to improve the accuracy and reduce the variability of the BSS-EPID dose-to-water conversion model. The correction may be applied as a preprocessing step

  14. Efficacy and Safety of MLC601 in the Treatment of Mild Cognitive Impairment: A Pilot, Randomized, Double-Blind, Placebo-Controlled Study

    Directory of Open Access Journals (Sweden)

    Hossein Pakdaman

    2017-05-01

    Full Text Available Background and Aim: Mild cognitive impairment (MCI is characterized by declined cognitive function greater than that expected for a person’s age. The clinical significance of this condition is its possible progression to dementia. MLC601 is a natural neuroprotective medication that has shown promising effects in Alzheimer disease. Accordingly, we conducted this randomized, double-blind, placebo-controlled study to evaluate the efficacy and safety of MLC601 in MCI patients. Methods: Seventy-two patients with a diagnosis of MCI were recruited. The included participants were randomly assigned to groups to receive either MLC601 or placebo. An evaluation of global cognitive function was performed at baseline as well as at 3-month and 6-month follow-up visits. Global cognitive function was assessed by Mini-Mental State Examination (MMSE and Alzheimer’s Disease Assessment Scale-cognitive subscale (ADAS-cog scores. Efficacy was evaluated by comparing global function scores between the 2 groups during the study period. Safety assessment included adverse events (AEs and abnormal laboratory results. Results: Seventy patients completed the study, 34 in the MLC601 group and 36 in the placebo group. The mean changes (±SD in cognition scores over 6 months in the MLC601 group were –2.26 (±3.42 for the MMSE and 3.82 (±6.16 for the ADAS-cog; in the placebo group, they were –2.66 (±3.43 for the MMSE and 4.41 (±6.66 for the ADAS-cog. The cognition changes based on both MMSE and ADAS-cog scores were statistically significant between the placebo and the MLC601 group (p < 0.001. Only 5 patients (14.7% reported minor AEs in the MLC601 group, the most commonly reported of which were gastrointestinal, none of them leading to patient withdrawal. Conclusion: MLC601 has shown promising efficacy and acceptable AEs in MCI patients.

  15. SU-E-T-128: Dosimetric Evaluation of MLC Modeling in Pinnacle V9.2 for Varian TrueBeam STx

    Energy Technology Data Exchange (ETDEWEB)

    Otageri, P; Grant, E; Maricle, S; Mathews, B [CARTI, Inc., Little Rock, AR (United States)

    2014-06-01

    Purpose: To evaluate the effects of MLC modeling after commissioning the Varian TrueBeam LINAC in Pinnacle version 9.2. Methods: Stepand-shoot IMRT QAs were investigated when we observed our measured absolute dose results using ion chamber (Capintec PR-05P) were uncharacteristically low; about 4–5% compared to doses calculated by Pinnacle{sup 3} (Phillips, Madison, WI). This problem was predominant for large and highly modulated head and neck (HN) treatments. Intuitively we knew this had to be related to shortcomings in the MLC modeling in Pinnacle. Using film QA we were able to iteratively adjust the MLC parameters. We confirmed results by re-testing five failed IMRT QA patients; and ion chamber measurements were verified in Quasar anthropomorphic phantom. Results: After commissioning the LINAC in Pinnacle version 9.2, the MLC transmission for 6X, 10X and 15X were 2.0%, 1.7% and 2.0%, respectively, and additional Interleaf leakage for all three energies was 0.5%. These parameters were obtained from profiles scanned with an Edge detector (Sun Nuclear, Melbourne, FL) during machine commissioning. A Verification testing with radiographic EDR2 film (Kodak, Rochester, NY) measurement was performed by creating a closed MLC leaf pattern and analyzing using RIT software (RIT, Colorado Springs, CO). This reduced MLC transmission for 6X, 10X and 15X to 0.7%, 0.9% and 0.9%, respectively; while increasing additional Interleaf leakage for all three energies to 1.0%. Conclusion: Radiographic film measurements were used to correct MLC transmission values for step and shoot IMRT fields used in Pinnacle version 9.2. After adjusting the MLC parameters to correlate with the film QA, there was still very good agreement between the Pinnacle model and commissioning data. Using the same QA methodology, we were also able to improve the beam models for the Varian C-series linacs, Novalis-Tx, and TrueBeam M-120 linacs.

  16. Obinutuzumab in chronic lymphocytic leukemia.

    Science.gov (United States)

    Dupuis, Jehan

    2015-09-01

    Obinutuzumab is the second next-generation monoclonal anti-CD20 antibody (after ofatumumab) to enter clinical practice in chronic lymphocytic leukemia. Its superiority in association with chlorambucil as compared with chlorambucil alone has led to its approval as a first-line treatment for chronic lymphocytic leukemia, for patients who are not candidates for a more intensive treatment.

  17. The lymphocyte transformation test in allergic contact dermatitis: New opportunities.

    Science.gov (United States)

    Popple, Amy; Williams, Jason; Maxwell, Gavin; Gellatly, Nichola; Dearman, Rebecca J; Kimber, Ian

    2016-01-01

    Allergic contact dermatitis (ACD) is driven by the activation and proliferation of allergen-specific memory T-lymphocytes and is currently diagnosed by patch testing with a selected panel of chemical allergens. The lymphocyte transformation test (LTT) can be used to monitor ex vivo T-lymphocyte responses to antigens, including contact allergens. The LTT is not viewed as being an alternative to patch testing, but it does seek to reflect experimentally skin sensitization to specific chemicals. The LTT is based on stimulation in vitro of antigen-driven T-lymphocyte proliferation. That is, exposure in culture of primed memory T-lymphocytes to the relevant antigen delivered in an appropriate configuration will provoke a secondary response that reflects the acquisition of skin sensitization. The technical aspects of this test and the utility of the approach for investigation of immune responses to contact allergens in humans are reviewed here, with particular emphasis on further development and refinement of the protocol. An important potential application is that it may provide a basis for characterizing those aspects of T-lymphocyte responses to contact allergens that have the greatest influence on skin sensitizing potency and this will be considered in some detail.

  18. Effects of cobalt and chromium ions on lymphocyte migration.

    Science.gov (United States)

    Baskey, Stephen J; Lehoux, Eric A; Catelas, Isabelle

    2017-04-01

    A T cell-mediated hypersensitivity reaction has been reported in some patients with CoCrMo-based implants. However, the role of cobalt and chromium ions in this reaction remains unclear. The objective of the present study was to analyze the effects of Co(2+) and Cr(3+) in culture medium, as well as the effects of culture supernatants of macrophages exposed to Co(2+) or Cr(3+) , on the migration of lymphocytes. The release of cytokines/chemokines by macrophages exposed to Co(2+) and Cr(3+) was also analyzed. The migration of murine lymphocytes was quantified using the Boyden chamber assay and flow cytometry, while cytokine/chemokine release by J774A.1 macrophages was measured by ELISA. Results showed an ion concentration-dependent increase in TNF-α and MIP-1α release and a decrease in MCP-1 and RANTES release. Migration analysis showed that the presence of Co(2+) (8 ppm) and Cr(3+) (100 ppm) in culture medium increased the migration of T lymphocytes, while it had little or no effect on the migration of B lymphocytes, suggesting that Co(2+) and Cr(3+) can stimulate the migration of T but not B lymphocytes. Levels of T lymphocyte migration in culture medium containing Co(2+) or Cr(3+) were not statistically different from those in culture supernatants of macrophages exposed to Co(2+) or Cr(3+) , suggesting that the effects of the ions and chemokines were not additive, possibly because of ion interference with the chemokines and/or their cognate receptors. Overall, results suggest that Co(2+) and Cr(3+) are capable of stimulating the migration of T (but not B) lymphocytes in the absence of cytokines/chemokines, and could thereby contribute to the accumulation of more T than B lymphocytes in periprosthetic tissues of some patients with CoCrMo-based implants. © 2016 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 35:916-924, 2017. © 2016 Orthopaedic Research Society. Published by Wiley Periodicals, Inc.

  19. Stimulation of rat B-lymphocyte proliferation by corticotropin-releasing factor.

    Science.gov (United States)

    McGillis, J P; Park, A; Rubin-Fletter, P; Turck, C; Dallman, M F; Payan, D G

    1989-07-01

    The mitogenic effect of corticotropin-releasing factor (CRF) on rat lymphocytes was investigated. When rat splenocytes were cultured for 48 hr with CFR, a dose-dependent increase in incorporation of 3H-thymidine (3H-Tdr) was observed, with a maximal response at 10 nM CRF. Comparison of the proliferative effect of CRF on enriched populations of B lymphocytes, T lymphocytes, or macrophages revealed that only B lymphocytes responded following treatment with CRF. When lymphocytes derived from different lymphoid tissues were compared, CRF had a greater proliferative effect on lymphocytes derived from gut-associated lymphoid tissue (mesenteric lymph nodes and Peyer's patches) than on lymphocytes from spleen or inguinal lymph nodes; CRF had no effect on thymocytes. Synthetic fragments of CRF were used to determine which portions of the peptide are recognized by lymphocytes. The C-terminal fragments alpha-helical CRF9-41 and CRF21-41 were as potent as native CRF in stimulating B-lymphocyte proliferation, whereas CRF1-20 did not stimulate proliferation. The activity of these peptides suggests that CRF stimulates lymphocyte proliferation by cellular recognition of structural determinants in the C-terminal one-half of the peptide.

  20. Sister chromatid exchange in human lymphocytes induced by propoxur following plant activation by Vicia faba.

    Science.gov (United States)

    Gómez-Arroyo, S; Calderón-Segura, M E; Villalobos-Pietrini, R

    1995-01-01

    Because the carbamate insecticide propoxur induced sister chromatid exchanges (SCE) in Vicia faba but was ineffective in producing SCE in lymphocytes in culture, it was hardly suspected that plant metabolism was involved. Experiments were conducted in which metabolic activation was afforded by Vicia faba roots, and SCE in human lymphocytes in vitro was used to assess cytogenetic damage. Several concentrations of propoxur (250, 500, 1,000, 1,500, and 2,000 ppm) were applied for 4 hr to the roots of Vicia faba. Extracts prepared from these treatments were added to the lymphocyte cultures and a significant increase of SCE frequencies with a concentration-response relationship could be detected. The lymphocyte proliferation kinetics and the proliferation rate index (PRI) were not affected (except in the highest concentration, of 2,000 ppm). This general behavior was in agreement with the presence of an enzymatic system (S10 fraction) in Vicia roots capable of metabolizing or activating the propoxur. With 2,000 ppm, cell necrosis was produced in Vicia; therefore, this extract did not induce SCE in lymphocytes. However, lymphocyte proliferation kinetics were delayed and PRI was significantly decreased. Ethanol, a promutagen activated by this plant, was applied directly to the lymphocyte cultures as a positive control, and the response was negative. On the other hand, the extracts of roots treated with ethanol increased the SCE to more than twice that of the negative control, but the lymphocyte proliferation kinetics and PRI were not affected.

  1. Reduced lymphocyte activation in space - Role of cell-substratum interactions

    Science.gov (United States)

    Gmuender, F. K.; Kiess, M.; Lee, J.; Cogoli, A.; Sonnenfeld, G.

    1992-01-01

    The effect of substratum adhesiveness on lymphocyte responsiveness was investigated by reducing and blocking cell adhesion with poly-HEMA in a simple on ground system. Cells grown on medium thick and thick poly-HEMA films were rounded in shape and displayed no signs of spreading. By contrast, on tissue culture plastic and very thin poly-HEMA films, they showed clear signs of spreading. The mitogenic response of lymphocytes grown on thick poly-HEMA films was reduced by up to 68 percent of the control (tissue culture plastic). Interferon gamma production was virtually nil when the cells were grown on the least adhesive substratum. These results show that activated lymphocytes need to anchor and spread prior to achieving an optimal proliferation response. It is concluded that decreased lymphocyte adhesion could contribute to the depressed in vitro lymphocyte responsiveness found in the microgravity conditions of space flight.

  2. MLC Libraries--A School Library's Journey with Students, Staff and Web 2.0 Technologies: Blogs, Wikis and E-Books--Where Are We Going Next?

    Science.gov (United States)

    Viner, Jane; Lucas, Amanda; Ricchini, Tracey; Ri, Regina

    2010-01-01

    This workshop paper explores the Web 2.0 journey of the MLC Libraries' teacher-librarians, librarian, library and audio visual technicians. Our journey was initially inspired by Will Richardson and supported by the School Library Association of Victoria (SLAV) Web 2.0 professional development program. The 12 week technological skills program…

  3. A Multistage Longitudinal Comparative (MLC) Design Stage II: Evaluation of the Changing Lives Program (CLP)--The Possible Selves Questionnaire-Qualitative Extensions (PSQ-QE)

    Science.gov (United States)

    Kortsch, Gabrielle; Kurtines, William M.; Montgomery, Marilyn J.

    2008-01-01

    The study reported in this paper, a Multistage Longitudinal Comparative (MLC) Design Stage II evaluation conducted as a planned preliminary efficacy evaluation (psychometric evaluation of measures, short-term controlled outcome studies, etc.) of the Changing Lives Program (CLP), provided evidence for the reliability and validity of qualitative…

  4. Development of an iterative reconstruction method to overcome 2D detector low resolution limitations in MLC leaf position error detection for 3D dose verification in IMRT

    NARCIS (Netherlands)

    Visser, Ruurd; Godart, J.; Wauben, D. J. L.; Langendijk, J. A.; van't Veld, A. A.; Korevaar, E. W.

    2016-01-01

    The objective of this study was to introduce a new iterative method to reconstruct multi leaf collimator (MLC) positions based on low resolution ionization detector array measurements and to evaluate its error detection performance. The iterative reconstruction method consists of a fluence model, a

  5. Laboratorial diagnosis of lymphocytic meningitis

    Directory of Open Access Journals (Sweden)

    Sérgio Monteiro de Almeida

    Full Text Available Meningitis is the main infectious central nervous system (CNS syndrome. Viruses or bacteria can cause acute meningitis of infectious etiology. The term "Aseptic Meningitis" denotes a clinical syndrome with a predominance of lymphocytes in the cerebrospinal fluid (CSF, with no common bacterial agents identified in the CSF. Viral meningitis is considered the main cause of lymphocyte meningitis. There are other etiologies of an infectious nature. CSF examination is essential to establish the diagnosis and to identify the etiological agent of lymphocytic meningitis. We examined CSF characteristics and the differential diagnosis of the main types of meningitis.

  6. Laboratorial diagnosis of lymphocytic meningitis

    Directory of Open Access Journals (Sweden)

    Sérgio Monteiro de Almeida

    2007-10-01

    Full Text Available Meningitis is the main infectious central nervous system (CNS syndrome. Viruses or bacteria can cause acute meningitis of infectious etiology. The term "Aseptic Meningitis" denotes a clinical syndrome with a predominance of lymphocytes in the cerebrospinal fluid (CSF, with no common bacterial agents identified in the CSF. Viral meningitis is considered the main cause of lymphocyte meningitis. There are other etiologies of an infectious nature. CSF examination is essential to establish the diagnosis and to identify the etiological agent of lymphocytic meningitis. We examined CSF characteristics and the differential diagnosis of the main types of meningitis.

  7. Microwave assisted sol-gel synthesis of high dielectric constant CCTO and BFN ceramics for MLC applications

    Directory of Open Access Journals (Sweden)

    Sonia

    2017-06-01

    Full Text Available Ba(Fe1/2Nb1/2O3 (BFN and CaCu3Ti4O12 (CCTO ceramic powders were synthesized by microwave assisted sol-gel synthesis technique and sintered at 1100°C and 1000°C, respectively. Calcination and sintering processes were carried out in a microwave furnace. Dielectric constant (εr~2450 and dielectric loss (tan δ~0.5 at frequency of 1 kHz and 20°C were observed for the BFN ceramic samples. Higher value of εr ~ 3600 and lower value of tan δ ~ 0.07 at frequency of 1 kHz and in 20-60°C temperature range for the CCTO ceramic samples suggested its utility for MLC applications. Sharp decrease of εr and sharp increase of tan δ at higher frequencies of BFN ceramic samples indicated the presence of Debye like relaxation.

  8. The use of intensity-modulated radiation therapy photon beams for improving the dose uniformity of electron beams shaped with MLC

    Energy Technology Data Exchange (ETDEWEB)

    Mosalaei, Homeira, E-mail: homeira.mosalaei@lhsc.on.ca [London Regional Cancer Program, London Health Science Centre, Ontario (Canada); Karnas, Scott [London Regional Cancer Program, London Health Science Centre, Ontario (Canada); University of Waterloo, Waterloo, Ontario (Canada); Shah, Sheel [University of Western Ontario, London, Ontario (Canada); Van Doodewaard, Sharon [McMaster University, Hamilton, Ontario (Canada); Foster, Tim [University of Western Ontario, London, Ontario (Canada); Chen, Jeff [London Regional Cancer Program, London Health Science Centre, Ontario (Canada); University of Waterloo, Waterloo, Ontario (Canada)

    2012-04-01

    Electrons are ideal for treating shallow tumors and sparing adjacent normal tissue. Conventionally, electron beams are collimated by cut-outs that are time-consuming to make and difficult to adapt to tumor shape throughout the course of treatment. We propose that electron cut-outs can be replaced using photon multileaf collimator (MLC). Two major problems of this approach are that the scattering of electrons causes penumbra widening because of a large air gap, and available commercial treatment planning systems (TPSs) do not support MLC-collimated electron beams. In this study, these difficulties were overcome by (1) modeling electron beams collimated by photon MLC for a commercial TPS, and (2) developing a technique to reduce electron beam penumbra by adding low-energy intensity-modulated radiation therapy (IMRT) photons (4 MV). We used blocks to simulate MLC shielding in the TPS. Inverse planning was used to optimize boost photon beams. This technique was applied to a parotid and a central nervous system (CNS) clinical case. Combined photon and electron plans were compared with conventional plans and verified using ion chamber, film, and a 2D diode array. Our studies showed that the beam penumbra for mixed beams with 90 cm source to surface distance (SSD) is comparable with electron applicators and cut-outs at 100 cm SSD. Our mixed-beam technique yielded more uniform dose to the planning target volume and lower doses to various organs at risk for both parotid and CNS clinical cases. The plans were verified with measurements, with more than 95% points passing the gamma criteria of 5% in dose difference and 5 mm for distance to agreement. In conclusion, the study has demonstrated the feasibility and potential advantage of using photon MLC to collimate electron beams with boost photon IMRT fields.

  9. Deleted in liver cancer 1 (DLC1 negatively regulates Rho/ROCK/MLC pathway in hepatocellular carcinoma.

    Directory of Open Access Journals (Sweden)

    Carmen Chak-Lui Wong

    Full Text Available AIMS: Deleted in liver cancer 1 (DLC1, a member of RhoGTPase activating protein (GAP family, is known to have suppressive activities in tumorigenicity and cancer metastasis. However, the underlying molecular mechanisms of how DLC1 suppresses cell motility have not been fully elucidated. Rho-kinase (ROCK is an immediate down-stream effector of RhoA in mediating cellular cytoskeletal events and cell motility. In the present study, we aimed to investigate the effects of DLC1 on Rho/ROCK signaling pathway in hepatocellular carcinoma (HCC. METHODOLOGY/PRINCIPAL FINDINGS: We demonstrated that DLC1 negatively regulated ROCK-dependent actomyosin contractility. From immunofluorescence study, we found that ectopic expression of DLC1 abrogated Rho/ROCK-mediated cytoskeletal reorganization including formation of stress fibers and focal adhesions. It also downregulated cortical phosphorylation of myosin light chain 2 (MLC2. These inhibitory events by DLC1 were RhoGAP-dependent, as RhoGAP-deficient mutant of DLC1 (DLC1 K714E abolished these inhibitory events. In addition, from western study, DLC1 inhibited ROCK-related myosin light chain phosphatase targeting unit 1 (MYPT1 phosphorylation at Threonine 853. By examining cell morphology under microscope, we found that ectopic expression of dominant-active ROCK released cells from DLC1-induced cytoskeletal collapse and cell shrinkage. CONCLUSION: Our data suggest that DLC1 negatively regulates Rho/ROCK/MLC2. This implicates a ROCK-mediated pathway of DLC1 in suppressing metastasis of HCC cells and enriches our understanding in the molecular mechanisms involved in the progression of hepatocellular carcinoma.

  10. Intensity- and energy-modulated electron radiotherapy by means of an xMLC for head and neck shallow tumors

    Energy Technology Data Exchange (ETDEWEB)

    Salguero, Francisco Javier; Palma, Bianey Atriana; Leal, Antonio [Departamento de FisiologIa Medica y Biofisica, Universidad de Sevilla (Spain); Arrans, Rafael [Hospital Universitario Virgen Macarena, Sevilla (Spain)], E-mail: alplaza@us.es

    2010-03-07

    The purpose of this paper is to assess the feasibility of delivering intensity- and energy-modulated electron radiation treatment (MERT) by a photon multileaf collimator (xMLC) and to evaluate the improvements obtained in shallow head and neck (HN) tumors. Four HN patient cases covering different clinical situations were planned by MERT, which used an in-house treatment planning system that utilized Monte Carlo dose calculation. The cases included one oronasal, two parotid and one middle ear tumors. The resulting dose-volume histograms were compared with those obtained from conventional photon and electron treatment techniques in our clinic, which included IMRT, electron beam and mixed beams, most of them using fixed-thickness bolus. Experimental verification was performed with plane-parallel ionization chambers for absolute dose verification, and a PTW ionization chamber array and radiochromic film for relative dosimetry. A MC-based treatment planning system for target with compromised volumes in depth and laterally has been validated. A quality assurance protocol for individual MERT plans was launched. Relative MC dose distributions showed a high agreement with film measurements and absolute ion chamber dose measurements performed at a reference point agreed with MC calculations within 2% in all cases. Clinically acceptable PTV coverage and organ-at-risk sparing were achieved by using the proposed MERT approach. MERT treatment plans, based on delivery of intensity-modulated electron beam using the xMLC, for superficial head and neck tumors, demonstrated comparable or improved PTV dose homogeneity with significantly lower dose to normal tissues. The clinical implementation of this technique will be able to offer a viable alternative for the treatment of shallow head and neck tumors.

  11. SU-E-T-475: An Accurate Linear Model of Tomotherapy MLC-Detector System for Patient Specific Delivery QA

    Energy Technology Data Exchange (ETDEWEB)

    Chen, Y; Mo, X; Chen, M; Olivera, G; Parnell, D; Key, S; Lu, W [21st Century Oncology, Madison, WI (United States); Reeher, M [21st Century Oncology, Naples, FL (United States); Galmarini, D [21st Century Oncology, Fort Myers, FL (United States)

    2014-06-01

    Purpose: An accurate leaf fluence model can be used in applications such as patient specific delivery QA and in-vivo dosimetry for TomoTherapy systems. It is known that the total fluence is not a linear combination of individual leaf fluence due to leakage-transmission, tongue-and-groove, and source occlusion effect. Here we propose a method to model the nonlinear effects as linear terms thus making the MLC-detector system a linear system. Methods: A leaf pattern basis (LPB) consisting of no-leaf-open, single-leaf-open, double-leaf-open and triple-leaf-open patterns are chosen to represent linear and major nonlinear effects of leaf fluence as a linear system. An arbitrary leaf pattern can be expressed as (or decomposed to) a linear combination of the LPB either pulse by pulse or weighted by dwelling time. The exit detector responses to the LPB are obtained by processing returned detector signals resulting from the predefined leaf patterns for each jaw setting. Through forward transformation, detector signal can be predicted given a delivery plan. An equivalent leaf open time (LOT) sinogram containing output variation information can also be inversely calculated from the measured detector signals. Twelve patient plans were delivered in air. The equivalent LOT sinograms were compared with their planned sinograms. Results: The whole calibration process was done in 20 minutes. For two randomly generated leaf patterns, 98.5% of the active channels showed differences within 0.5% of the local maximum between the predicted and measured signals. Averaged over the twelve plans, 90% of LOT errors were within +/−10 ms. The LOT systematic error increases and shows an oscillating pattern when LOT is shorter than 50 ms. Conclusion: The LPB method models the MLC-detector response accurately, which improves patient specific delivery QA and in-vivo dosimetry for TomoTherapy systems. It is sensitive enough to detect systematic LOT errors as small as 10 ms.

  12. SU-E-T-363: Error Detection Comparison of EPID and MLC Log File Based IMRT QA Systems

    Energy Technology Data Exchange (ETDEWEB)

    Defoor, D; Obeidat, M; Linden, P; Kirby, N; Papanikolaou, N; Stathakis, S [University of Texas Health Science Center at San Antonio, San Antonio, TX (United States); Mavroidis, P [University of North Carolina, Chapel Hill, NC (United States)

    2015-06-15

    Purpose: In this study we will compare the ability of three QA methods (Delta4, MU-EPID, Dynalog QA) to detect specific errors. Methods: A Varian Novalis Tx with a HD120 MLC and aS1000 Electronic Portal Imaging Device (EPID) was used in our study. Multi-leaf collimator (MLC) errors, gantry angle and dose errors were introduced into 5 volumetric arc therapy (VMAT) plans. 3D dose distributions calculated with data from the EPID and Dynalog QA methods were compared with the planned dose distribution. The gamma passing percentages as well as percentage error of planning target volume (PTV) dose were used for passing determination. Baselines for gamma passing percentages and PTV dose were established by measuring the original plan 5 times consecutively. Standard passing thresholds as well as thresholds derived from receiver operator characteristic (ROC) analysis and 2 standard deviation (SD) criteria were used. Results: When applying the standard 95% pass rate at 3%/3mm gamma analysis 14, 21 and 8 of 30 errors were detected by the Delta4, MU-EPID and Dynalog QA methods respectively. Thresholds set at 2 SD from our base line measurements resulted in the detection of 18, 9 and 14 of 30 errors for the Delta4, MU-EPID and Dynalog QA methods respectively. When using D2 of the PTV as a metric the Dynalog QA detected 20 of 30 errors while the EPID method detected 14 of 30 errors. Using D98 of the PTV, Dynalog QA detected 13 of 30 while the EPID detected 3 of 30 errors. Conclusion: Although MU-EPID detected the most errors at the standard 95% cutoff it also produced the most false detections in the baseline data. The Dynalog QA was the most effective when the ROC adjusted passing threshold was used. D2 was more effective as a metric for detecting errors than D98.

  13. Genotoxic effects of bistratene A on human lymphocytes.

    Science.gov (United States)

    Rojas, E; Valverde, M; Vega, L; Salvador, A; Ramirez, P; Herrera, L A; Watters, D; Lavin, M F; Ostrosky-Wegman, P

    1996-03-01

    Bistratene A, a toxin isolated from the colonial ascidian Lissoclinum bistratum causes a decrease in mitotic index and retardation of lymphocyte proliferation kinetics when it is added at 48 h to 72-h human lymphocyte cultures. In the same cultures, the incidence of sister chromatid exchanges was not altered by this compound. We also observed an increase in the number of polyploid cells in the cultures, and alterations of the beta-tubulin organization by immunocytochemistry with an antibody against beta-tubulin. Bistratene A induces DNA damage in a dose-dependent fashion in leukocytes, as measured by the alkaline single cell gel electrophoresis assay. These results show that bistratene A interferes with microtubule assembly, is cytotoxic and cytostatic, and that it causes DNA damage.

  14. 中华鳖太湖种群淋巴细胞培养及染色体核型分析%The Lymphocytic Cell Culture and Chromosome Karyotype of Trionyx sinensis in Taihu Lake Population

    Institute of Scientific and Technical Information of China (English)

    熊良伟; 王建国; 封琦; 黄爱军

    2013-01-01

    In order to provide the logical proofs for germplasm research and chromosomal manipulation of Trionyx sinensis. The karyotypes of the chromosomes of Trionyx sinensis in Taihu Lake were examined by lymphocytic cell cultivation-air drying-Giemsa staining method. The results showed that the chromosome karyotype of T. Sinensis in Taihu Lake population could be obtained through the lymphocytic cell cultivation method. There were 33 pairs chromosome including 6 pairs of large chromosome and 27 pairs of smaller chromosome.%为给中华鳖种质研究和染色体操作提供理论依据.以中华鳖太湖种群为材料,采用淋巴细胞培养、空气干燥法制片、Giemsa染色,对中华鳖染色体核型进行了分析.结果表明:通过淋巴细胞培养法可以得到良好的染色体制片;中华鳖太湖种群具有33对染色体,其中6对是大型染色体,27对是小型染色体.

  15. Initiation of lymphocyte DNA synthesis.

    Science.gov (United States)

    Coffman, F D; Fresa, K L; Cohen, S

    1991-01-01

    The initiation of DNA replication in T lymphocytes appears to be regulated by two distinct activities: one associated with proliferation which mediates initiation, and another associated with quiescence which blocks initiation. Activated lymphocytes and proliferating lymphoid cell lines produce an activity, termed ADR, which can initiate DNA replication in isolated, quiescent nuclei. ADR is heat-labile, has protease activity or interacts closely with a protease, and is distinct from the DNA polymerases. ADR activity is absent in quiescent lymphocytes and appears in mitogen-stimulated lymphocytes after IL-2 binding. The generation of active ADR appears to be mediated by phosphorylation of a precursor which is present in resting cells. Nuclei from mitogen-unresponsive lymphocytes fail to initiate DNA replication in response to ADR, of potential importance in the age-related decline of immunity. Quiescent lymphocytes lack ADR and synthesize an ADR-inhibitory activity. The ADR inhibitor is a heat-stable protein which suppresses the initiation of DNA synthesis, but is ineffective at suppressing elongation once DNA strand replication has begun. Nuclei from several neoplastic cell lines fail to respond to the ADR inhibitor, which may play a role in the continuous proliferation of these cells. At least one of these neoplastic cell lines produces both ADR and an inhibitory factor. These findings suggest that the regulation of proliferation is dependent on the balance between activating and inhibitory pathways.

  16. 斑鳜轻链3基因(MLC3)cDNA的克隆及纵向表达分析%cDNA Cloning and Vertical Expression Analysis of the Myosin Light Chain 3 ( MLC3 ) Gene in Siniperca Scherzeri

    Institute of Scientific and Technical Information of China (English)

    农小献; 赵发兰; 宾石玉; 陈敦学; 刘希良; 史竸; 褚武英

    2011-01-01

    提取斑鳜肌肉组织总RNA,逆转录得到cDNA,然后利用常规PCR法扩增得到斑鳜肌球蛋白轻链3基因(MLC3)序列.斑鳜MLC3基因cDNA序列的全长为532 bp,编码150个氨基酸.通过PROSITE tools软件预测显示该序列具有2个EF-手相结构.通过实时荧光定量PCR法对斑鳜MLC3纵向表达进行分析发现,MLC3在斑鳜背肌的近头部、中部和近尾部都有表达,近头部和中部的表达量差异不显著,近尾部表达量显著高于近头部和中部.%RNA of Siniperca scherzeri was isolated from muscle tissue, applied differential display reversed transcripted PCR (DDRT-PCR) technique to get cDNA sequence of MLC3, which is 532 bp in length, and encodes a 150 amino acid peptide.It is found that it contains two EF-hand motifs by using the indicating PROSITE tools software.Longitudinal expression of mandarin fish was studied by using Real-time Quantitative PCR showed that the MLC3 mRNA was expressed along longitudinal axis with unsignificant between the anterior and middle muscle positions, but significant variation between the posterior and anterior, middle muscle positions.

  17. T lymphocyte insensitivity to corticosteroids in chronic obstructive pulmonary disease

    Directory of Open Access Journals (Sweden)

    Kaur Manminder

    2012-03-01

    Full Text Available Abstract Background There are increased numbers of activated lymphocytes in the lungs of chronic obstructive pulmonary disease (COPD patients. The clinical benefits of corticosteroids in COPD patients are limited. Our hypothesis is that lymphocytes play a role in this corticosteroid insensitivity. Objectives To investigate the effects of the corticosteroid dexamethasone on lung lymphocyte cytokine production from patients with COPD compared to controls. Methods Cultured airway lymphocytes obtained by bronchoscopy from healthy non-smokers (HNS, smokers (S and COPD patients were stimulated with phytohaemagglutinin (PHA & phorbol myristate acetate (PMA, +/- dexamethasone. Supernatants were assayed for interleukin (IL-2 and interferon (IFNγ. Immunofluoresence was used to analyse changes in CD8 glucocorticoid receptor (GRα and GRβ expression. Results The inhibition of PHA/PMA stimulated IFNγ production by dexamethasone was reduced in COPD patients compared to HNS (p p Conclusions IFNγ production from COPD airway lymphocytes is corticosteroid insensitive. This phenomenon may be important in the poor clinical response often observed with corticosteroids.

  18. Regulation of PTS gene expression by the homologous transcriptional regulators, Mlc and NagC, in Escherichia coli (or how two similar repressors can behave differently).

    Science.gov (United States)

    Plumbridge, J

    2001-07-01

    NagC and Mlc are paralogous transcriptional repressors in E.coli. Unexpectedly they possess almost identical amino acid sequences in their helix-turn-helix (H-T-H), DNA binding motif and they bind to very similar consensus operator targets. Binding to each others sites can be demonstrated in vitro but no cross regulation can be detected in vivo with physiological amounts of the two proteins. Although both proteins are involved in regulating the expression of PTS genes, the characteristics of their repression and induction are very different. NagC is a dual-function, activator-repressor which co-ordinates the metabolism of the amino sugars, N-acetylglucosamine (GlcNAc) and glucosamine, by repressing the divergent nagE-BA operons and by activating the glmUS operon. Repression (and activation) by NagC requires that NagC binds simultaneously to two operators, thus forming a DNA loop. This chelation effect allows use of lower affinity sites which would not individually bind the repressor. The specific inducer for NagC is GlcNAc-6-P, the product of GlcNAc transport by the PTS and a key compound in amino sugar metabolism. Mlc represses several genes implicated in the uptake of glucose; ptsG, ptsHI and manXYZ, and malT, the activator of the mal regulon. Glucose behaves like the inducer but growth on glucose only produces an overall increase in expression for ptsG and ptsHI. All Mlc repressed genes are also controlled by cAMP/CAP, so that glucose affects their transcription in two opposing ways: increasing expression by acting as the inducer for Mlc but decreasing expression by lowering the cAMP/CAP concentration. The Mlc protein is not directly responsive to glucose per se but to the activity status of the PTS. Displacement of Mlc from its binding sites occurs during growth on glucose and other PTS sugars and involves sequestration of the repressor to membranes by binding to dephosphorylated PtsG.

  19. Isolation of Lymphocytes and Their Innate Immune Characterizations from Liver, Intestine, Lung and Uterus

    Institute of Scientific and Technical Information of China (English)

    Jianhong Zhang; Zhongjun Dong; Rongbin Zhou; Deming Luo; Haiming Wei; Zhigang Tian

    2005-01-01

    In steady-state conditions, the number and distribution of lymphocyte populations are under homeostatic control.New lymphocytes are continuously produced in primary and secondary lymphoid organs and then achieve immune-competence within different tissues, and they must challenge with resident cells for survival. The first step in the study of tissue lymphoid cells is their isolation in intact and viable form appropriate for establishment of in vitro culture systems. For reasons of simplicity, cell purity, cell yields and various purposes, lymphocytes obtained from different tissues in different labs were subjected to diverse protocols. To fully elucidate the nature of the local immune system as well as to adequately study the innate role of lymphocytes in liver, intestine, lung and uterus, we briefly reviewed the characterization of resident lymphocytes, and additional information on those cells from non-lymphoid tissues by using the recommended operation procedure was also presented.

  20. Microscopic structures of Japanese lamprey peripheral blood cells and culture of lymphocyte-like cells in vitro%日本七鳃鳗外周血细胞显微结构及类淋巴细胞体外培养

    Institute of Scientific and Technical Information of China (English)

    逢越; 李庆伟

    2012-01-01

    The mierostructure of Japanese lamprey (Lampetra japonica) peripheral blood cells and the cultivation methods of lymphocyte-like cells were explored. The peripheral blood cells of L. japonica were stained by Wright's and Giemsa's staining methods and five major cell types including erythrocyte, lymphocyte, neutrophil granulocyte, monocyte and eosinophil granulocyte, but not basophil were recognized under light microscope. The lymphocyte-like cells were separated successfully with Ficoll-Paque centrifugation and flow cytometry and were cultured in L15 medium with 20% foetal bovine serum (FBS) including 5% L. japonica FBS and 0.46% NaCI. The optimal temperature of cells is 18 ℃ and the optimal pH of cells is 6.8-7.0. They can develop attachment as a sheet of cells after 1 hour. Cells have a good condition for nearly a month. The establishment of primary cell cultures of lymphocyte-like cells from L. japonica provides a useful tool for the longevity of the culture and the investigation in cellular biology.%以日本七鳃鳗(Lampetrajaponica)为研究对象,观察其外周血细胞显微结构和探讨类淋巴细胞原代培养条件。采用Wright氏和Giemsa氏染色法对日本七鳃鳗外周血液有形成分进行显微观察,可鉴别出红细胞、淋巴细胞、中性粒细胞、单核细胞、嗜酸性粒细胞:未发现嗜碱性粒细胞。通过Ficoll密度梯度离心法和流式细胞仪分选获得高纯度的七鳃鳗类淋巴细胞。经条件优化后确定最适培养液L15+20%FBS(5%七鳃鳗血清)+0.46%NaCl,在18℃,pH6.8~7.0的条件下对日本七鳃鳗类淋巴细胞进行体外培养,大多数类淋巴细胞半贴壁生长,细胞状态较好,最长可以存活近一个月。类淋巴细胞的原代培养为日本七鳃鳗细胞体外培养的深入研究提供了实验依据。

  1. Kinetic analysis of interleukin 2 (IL-2) production and expression of IL-2 receptors by uraemic and normal lymphocytes

    DEFF Research Database (Denmark)

    Langhoff, E; Hofmann, B; Ladefoged, J

    1987-01-01

    The in vitro immune response of lymphocytes from uraemic patients was studied by comparing the in vitro kinetics of interleukin 2 (IL-2) production, the mitogen-induced proliferative response, and the expression of IL-2 receptors by T lymphocytes. The IL-2 production in 26 uraemic cell cultures...

  2. Expansion of activated lymphocytes obtained from renal cell carcinoma in an automated hollow fiber bioreactor.

    Science.gov (United States)

    Hillman, G G; Wolf, M L; Montecillo, E; Younes, E; Ali, E; Pontes, J E; Haas, G P

    1994-01-01

    Immunotherapy using IL-2 alone or combined with activated lymphocytes has been promising for metastatic renal cell carcinoma. Cytotoxic lymphocytes can be isolated from tumors, expanded in vitro with IL-2, and adoptively transferred back into the tumor-bearing host. These cells can also be transduced with the genes coding for cytokines for local delivery to tumor sites. A major drawback in adoptive immunotherapy is the cumbersome and expensive culture technology associated with the growth of large numbers of cells required for their therapeutic effect. To reduce the cost, resources, and manpower, we have developed the methodology for lymphocyte activation and expansion in the automated hollow fiber bioreactor IMMUNO*STAR Cell Expander (ACT BIOMEDICAL, INC). Tumor Infiltrating Lymphocytes (TIL) isolated from human renal cell carcinoma tumor specimens were inoculated at a number of 10(8) cells in a small bioreactor of 30 ml extracapillary space volume. We have determined the medium flow rates and culture conditions to obtain a significant and repeated expansion of TIL at weekly intervals. The lymphocytes cultured in the bioreactor demonstrated the same phenotype and cytotoxic activity as those expanded in parallel in tissue culture plates. Lymphocyte expansion in the hollow fiber bioreactor required lower volumes of medium, human serum, IL-2 and minimal labor. This technology may facilitate the use of adoptive immunotherapy for the treatment of refractory malignancies.

  3. Improvements in dose accuracy delivered with static-MLC IMRT on an integrated linear accelerator control system

    Energy Technology Data Exchange (ETDEWEB)

    Li Ji; Wiersma, Rodney D.; Stepaniak, Christopher J.; Farrey, Karl J.; Al-Hallaq, Hania A. [Department of Radiation and Cellular Oncology, University of Chicago, 5758 South Maryland Avenue, MC9006, Chicago, Illinois 60637 (United States)

    2012-05-15

    Purpose: Dose accuracy has been shown to vary with dose per segment and dose rate when delivered with static multileaf collimator (SMLC) intensity modulated radiation therapy (IMRT) by Varian C-series MLC controllers. The authors investigated the impact of monitor units (MUs) per segment and dose rate on the dose delivery accuracy of SMLC-IMRT fields on a Varian TrueBeam linear accelerator (LINAC), which delivers dose and manages motion of all components using a single integrated controller. Methods: An SMLC sequence was created consisting of ten identical 10 x 10 cm{sup 2} segments with identical MUs. Beam holding between segments was achieved by moving one out-of-field MLC leaf pair. Measurements were repeated for various combinations of MU/segment ranging from 1 to 40 and dose rates of 100-600 MU/min for a 6 MV photon beam (6X) and dose rates of 800-2400 MU/min for a 10 MV flattening-filter free photon (10XFFF) beam. All measurements were made with a Farmer (0.6 cm{sup 3}) ionization chamber placed at the isocenter in a solid-water phantom at 10 cm depth. The measurements were performed on two Varian LINACs: C-series Trilogy and TrueBeam. Each sequence was delivered three times and the dose readings for the corresponding segments were averaged. The effects of MU/segment, dose rate, and LINAC type on the relative dose variation ({Delta}{sub i}) were compared using F-tests ({alpha} = 0.05). Results: On the Trilogy, large {Delta}{sub i} was observed in small MU segments: at 1 MU/segment, the maximum {Delta}{sub i} was 10.1% and 57.9% at 100 MU/min and 600 MU/min, respectively. Also, the first segment of each sequence consistently overshot ({Delta}{sub i} > 0), while the last segment consistently undershot ({Delta}{sub i} < 0). On the TrueBeam, at 1 MU/segment, {Delta}{sub i} ranged from 3.0% to 4.5% at 100 and 600 MU/min; no obvious overshoot/undershoot trend was observed. F-tests showed statistically significant difference [(1 - {beta}) =1.0000] between the

  4. Dosimetric and delivery efficiency investigation for treating hepatic lesions with a MLC-equipped robotic radiosurgery–radiotherapy combined system

    Energy Technology Data Exchange (ETDEWEB)

    Jin, Lihui, E-mail: lihui.jin@fccc.edu; Price, Robert A.; Wang, Lu; Meyer, Joshua; Fan, James; Charlie Ma, Chang Ming [Department of Radiation Oncology, Fox Chase Cancer Center, 333 Cottman Avenue, Philadelphia, Pennsylvania 19111 (United States)

    2016-02-15

    Purpose: The CyberKnife M6 (CK-M6) Series introduced a multileaf collimator (MLC) for extending its capability from stereotactic radiosurgery/stereotactic radiotherapy (SBRT) to conventionally fractionated radiotherapy. This work is to investigate the dosimetric quality of plans that are generated using MLC-shaped beams on the CK-M6, as well as their delivery time, via comparisons with the intensity modulated radiotherapy plans that were clinically used on a Varian Linac for treating hepatic lesions. Methods: Nine patient cases were selected and divided into three groups with three patients in each group: (1) the group-one patients were treated conventionally (25 fractions); (2) the group-two patients were treated with SBRT-like hypofractionation (5 fractions); and (3) the group-three patients were treated similar to group-one patients, but with two planning target volumes (PTVs) and two different prescription dose levels correspondingly. The clinically used plans were generated on the ECLIPSE treatment planning system (TPS) and delivered on a Varian Linac (E-V plans). The multiplan (MP) TPS was used to replan these clinical cases with the MLC as the beam device for the CK-M6 (C-M plans). After plans were normalized to the same PTV dose coverage, comparisons between the C-M and E-V plans were performed based on D{sub 99%} (percentage of prescription dose received by 99% of the PTV), D{sub 0.1cm{sup 3}} (the percentage of prescription dose to 0.1 cm{sup 3} of the PTV), and doses received by critical structures. Then, the delivery times for the C-M plans will be obtained, which are the MP TPS generated estimations assuming having an imaging interval of 60 s. Results: The difference in D{sub 99%} between C-M and E-V plans is +0.6% on average (+ or − indicating a higher or lower dose from C-M plans than from E-V plans) with a range from −4.1% to +3.8%, and the difference in D{sub 0.1cm{sup 3}} was −1.0% on average with a range from −5.1% to +2.9%. The PTV

  5. Preliminary studies for implementation of a MCL quality control using EPID (Portal Dosimetry); Estudos preliminares para implementacao de um controle de qualidade de MLC com o uso do EPID (Portal Dosimetry)

    Energy Technology Data Exchange (ETDEWEB)

    Mattos, Fabio R.; Furnari, Laura [Universidade de Sao Paulo (USP), Sao Paulo, SP (Brazil). Faculdade de Medicina

    2016-07-01

    A Quality Control (CQ) to ensure the expected performance of a Multileaf Collimator System (MLC) is essential for delivering dose in a safety and appropriate way. The time required for equipment control and dosimetry may be lowered when the Electronic Portal Image Device (EPID) is used. The aim of this paper was to check the resolution limits of the detection system for IMRT mode, and to do the analysis of three tests of MLC performance: Picket Fence, Slinding GAP, MLC versus Gantry. A Varian iX Clinac equipped with an 80 leaf Millennium MLC and with amorphous silicon based EPID (aS1000) was use. The EPID proved Effective, where errors up to 0,5 mm can be detected. Information about interleaf transmissions, dose profile and gravity influence in the leaf banks also were included. (author)

  6. Immunomodulatory effects of oral antidiabetic drugs in lymphocyte cultures from patients with type 2 diabetes Efeito imunomodulador de hipoglicemiantes orais em cultura de linfócitos de pacientes com diabetes tipo 2

    Directory of Open Access Journals (Sweden)

    Karina Faccio Mello

    2011-02-01

    Full Text Available INTRODUCTION AND OBJECTIVE: It has been suggested that type 2 diabetes is an inflammatory response manifestation. The main drugs used to treat type 2 diabetes are sulphonylureas and biguanides. The aim of this study was to demonstrate the modulatory effects of oral hypoglycemic drugs (chlorpropamide and metformin on lymphocyte proliferation in vitro and ex vivo. METHODS: Peripheral blood mononuclear cells were isolated from human blood by gradient centrifugation. T-lymphocytes were stimulated by phytohemagglutinin (PHA and oral hypoglycemic drugs. RESULTS: In both in vitro and ex vivo experiments, there was a reduction in cell proliferation after treatment with oral hypoglycemic drugs. When both drugs were used in combination, a high level of cytotoxicity was observed, which made analysis of immunomodulatory effects unfeasible. DISCUSSION AND CONCLUSION: We demonstrated that diabetes itself may reduce cell proliferation significantly when stimulated by PHA, which may indicate that diabetic patients have difficulties in promoting an efficient inflammatory response. Moreover, the use of oral hypoglycemic drugs may aggravate this situation.INTRODUÇÃO E OBJETIVOS: Tem sido sugerido que o diabetes mellitus tipo 2 (DM2 é uma manifestação da resposta inflamatória. As principais drogas utilizadas no tratamento do DM2 são as sulfonilureias e as biguanidas. O objetivo deste trabalho é demonstrar os efeitos moduladores na proliferação de linfócitos causada pelos hipoglicemiantes orais (clorpropamida e metformina, in vitro e ex vivo. MÉTODOS: Células mononucleares de sangue periférico foram isoladas de seres humanos por gradiente de centrifugação. Os linfócitos T foram estimulados com fito-hemaglutinina (PHA e hipoglicemiantes. RESULTADOS: Nos experimentos in vitro e ex vivo, mostramos a redução da proliferação celular quando do tratamento com drogas hipoglicemiantes orais. Quando as drogas foram utilizadas em combinação, foi

  7. Effects of tertiary MLC configuration on secondary neutron spectra from 18 MV x-ray beams for the Varian 21EX linear accelerator.

    Science.gov (United States)

    Howell, Rebecca M; Kry, Stephen F; Burgett, Eric; Followill, David; Hertel, Nolan E

    2009-09-01

    The effect of the jaw configuration and the presence and configuration of the tertiary multileaf collimator (MLC) on the secondary neutron spectra for an 18 MV Varian 21EX linear accelerator (linac) is investigated in detail. The authors report the measured spectra for four collimator (jaw-and-MLC) configurations. These configurations represent the extreme settings of the jaws and MLC and should therefore describe the range of possible fluence and spectra that may be encountered during use of this linac. In addition to measurements, a Monte Carlo model was used to simulate the four collimator configurations and calculate the energy spectra and fluence at the same location as it was measured. The Monte Carlo model was also used to calculate the sources of neutron production in the linac head for each collimator configuration. They found that photoneutron production in the linac treatment head is dominated by the order in which the primary photon beam intercepts the high-Z material. The primary collimator, which has the highest position in the linac head (in a fixed location), is the largest source of secondary neutrons. Thereafter, the collimator configuration plays a role in where the neutrons originate. For instance, if the jaws are closed, they intercept the beam and contribute substantially to the secondary neutron production. Conversely, if the jaws are open, the MLC plays a larger role in neutron production (assuming, of course, that it intercepts the beam). They found that different collimator configurations make up to a factor of 2 difference in the ambient dose equivalent.

  8. Melatonin Attenuates Aortic Endothelial Permeability and Arteriosclerosis in Streptozotocin-Induced Diabetic Rats: Possible Role of MLCK- and MLCP-Dependent MLC Phosphorylation.

    Science.gov (United States)

    Tang, Song-tao; Su, Huan; Zhang, Qiu; Tang, Hai-qin; Wang, Chang-jiang; Zhou, Qing; Wei, Wei; Zhu, Hua-qing; Wang, Yuan

    2016-01-01

    The development of diabetic macrovascular complications is a multifactorial process, and melatonin may possess cardiovascular protective properties. This study was designed to evaluate whether melatonin attenuates arteriosclerosis and endothelial permeability by suppressing the myosin light-chain kinase (MLCK)/myosin light-chain phosphorylation (p-MLC) system via the mitogen-activated protein kinase (MAPK) signaling pathway or by suppressing the myosin phosphatase-targeting subunit phosphorylation (p-MYPT)/p-MLC system in diabetes mellitus (DM). Rats were randomly divided into 4 groups, including control, high-fat diet, DM, and DM + melatonin groups. Melatonin was administered (10 mg/kg/d) by gavage for 12 weeks. The DM significantly increased the serum fasting blood glucose and lipid levels, as well as insulin resistance and endothelial dysfunction, which were attenuated by melatonin therapy to various extents. Importantly, the aortic endothelial permeability was significantly increased in DM rats but was dramatically reversed following treatment with melatonin. Our findings further indicated that hyperglycemia and hyperlipidemia enhanced the expressions of MLCK, p-MYPT, and p-MLC, which were partly associated with decreased membrane type 1 expression, increased extracellular signal-regulated kinase (ERK) phosphorylation, and increased p38 expression. However, these changes in protein expression were also significantly reversed by melatonin. Thus, our results are the first to demonstrate that the endothelial hyperpermeability induced by DM is associated with increased expressions of MLCK, p-MYPT, and p-MLC, which can be attenuated by melatonin at least partly through the ERK/p38 signaling pathway.

  9. SU-E-T-418: Explore the Sensitive of the Planar Quality Assurance to the MLC Error with Different Beam Complexity in Intensity-Modulate Radiation Therapy

    Energy Technology Data Exchange (ETDEWEB)

    Wang, J; Peng, J; Xie, J; Hu, W [Fudan university shanghai cancer center, Shanghai (China)

    2015-06-15

    Purpose: The purpose of this study is to investigate the sensitivity of the planar quality assurance to MLC errors with different beam complexities in intensity-modulate radiation therapy. Methods: sixteen patients’ planar quality assurance (QA) plans in our institution were enrolled in this study, including 10 dynamic MLC (DMLC) IMRT plans measured by Portal Dosimetry and 6 static MLC (SMLC) IMRT plans measured by Mapcheck. The gamma pass rate was calculated using vender’s software. The field numbers were 74 and 40 for DMLC and SMLC, respectively. A random error was generated and introduced to these fields. The modified gamma pass rate was calculated by comparing the original measured fluence and modified fields’ fluence. A decreasing gamma pass rate was acquired using the original gamma pass rate minus the modified gamma pass rate. Eight complexity scores were calculated in MATLAB based on the fluence and MLC sequence of these fields. The complexity scores include fractal dimension, monitor unit of field, modulation index, fluence map complexity, weighted average of field area, weighted average of field perimeter, and small aperture ratio ( <5cm{sup 2} and <50cm{sup 2}). The Spearman’s rank correlation coefficient was implemented to analyze the correlation between these scores and decreasing gamma rate. Results: The relation between the decreasing gamma pass rate and field complexity was insignificant for most complexity scores. The most significant complexity score was fluence map complexity for SMLC, which have ρ =0.4274 (p-value=0.0063). For DMLC, the most significant complex score was fractal dimension, which have ρ=−0.3068 (p-value=0.0081). Conclusions: According to the primarily Result of this study, the sensitivity gamma pass rate was not strongly relate to the field complexity.

  10. In vitro genotoxicity of pyridine in human lymphocytes.

    Science.gov (United States)

    Emelnsia, Aida D; Rather, Irfan A

    2016-05-01

    This work was carried out to study the genotoxicity of pyridine in vitro on human leucocyte culture. Cyclophosphamide, a well-known carcinogen was used as positive control. The four different concentrations of pyridine and cyclophosphamide showed breaks and pulverization of chromosomes in dose dependent manner. Higher number of pulverization was observed with higher concentration of pyridine (3.25μg/mL). Based on this data, our results confirm that both pyridine and its precursor showed genotoxicity against human lymphocytes.

  11. Genotoxicity of food preservative sodium sorbate in human lymphocytes in vitro.

    Science.gov (United States)

    Mamur, Sevcan; Yüzbaşıoğlu, Deniz; Unal, Fatma; Aksoy, Hüseyin

    2012-10-01

    The genotoxic effects of antimicrobial food additive sodium sorbate (SS) was assessed by using chromosome aberrations (CAs), sister-chromatid exchanges (SCEs), and micronucleus (MN) in cultured human lymphocytes and comet assay in isolated human lymphocytes. Lymphocytes were treated with four concentrations (100, 200, 400 and 800 μg/ml) of SS as well as a negative (sterile distilled water) and a positive control (Mitomycin-C: MMC for cultured lymphocytes and H(2)O(2) for isolated lymphocytes). The result of this study indicated that SS increased the frequency of CAs at both 24 and 48 h period compared to control. When gaps were included, this increase was significant at 200, 400 and 800 μg/ml concentrations at 24 h and, at all concentrations at 48 h treatment time. When gaps were excluded, this increase was significant at only 800 μg/ml concentration at both 24 and 48 h treatments. In addition, SS increased SCEs/cell and MN frequency at 400 and 800 μg/ml concentrations at both 24 and 48 h compared to negative control. Furthermore, this additive caused DNA damage at all concentrations in isolated human lymphocytes after 1 h in vitro exposure. The present results show that SS is genotoxic to the human peripheral blood lymphocytes in vitro at the highest concentrations.

  12. Airborne particulate matter in vitro exposure induces cytoskeleton remodeling through activation of the ROCK-MYPT1-MLC pathway in A549 epithelial lung cells.

    Science.gov (United States)

    Chirino, Yolanda I; García-Cuellar, Claudia María; García-García, Carlos; Soto-Reyes, Ernesto; Osornio-Vargas, Álvaro Román; Herrera, Luis A; López-Saavedra, Alejandro; Miranda, Javier; Quintana-Belmares, Raúl; Pérez, Irma Rosas; Sánchez-Pérez, Yesennia

    2017-03-06

    Airborne particulate matter with an aerodynamic diameter ≤10μm (PM10) is considered a risk factor for the development of lung cancer. Little is known about the cellular mechanisms by which PM10 is associated with cancer, but there is evidence that its exposure can lead to an acquired invasive phenotype, apoptosis evasion, inflammasome activation, and cytoskeleton remodeling in lung epithelial cells. Cytoskeleton remodeling occurs through actin stress fiber formation, which is partially regulated through ROCK kinase activation, we aimed to investigate if this protein was activated in response to PM10 exposure in A549 lung epithelial cells. Results showed that 10μg/cm(2) of PM10 had no influence on cell viability but increased actin stress fibers, cytoplasmic ROCK expression, and phosphorylation of myosin phosphatase-targeting 1 (MYPT1) and myosin light chain (MLC) proteins, which are targeted by ROCK. The inhibition of ROCK prevented actin stress fiber formation and the phosphorylation of MYPT1 and MLC, suggesting that PM10 activated the ROCK-MYPT1-MLC pathway in lung epithelial cells. The activation of ROCK1 has been involved in the acquisition of malignant phenotypes, and its induction by PM10 exposure could contribute to the understanding of PM10 as a risk factor for cancer development through the mechanisms associated with invasive phenotype.

  13. Reactivity of eleven anti-human leucocyte monoclonal antibodies with lymphocytes from several domestic animals

    DEFF Research Database (Denmark)

    Aasted, Bent; Blixenkrone-Møller, Merete; Larsen, Else Bang

    1988-01-01

    Nine commercially available monoclonal antibodies and two monoclonal antibodies from The American Type Culture Collection, raised against various human leucocyte surface antigens, were tested on lymphocytes from cow, sheep, goat, swine, horse, cat, dog, mink, and rabbit as well as man. Four...... antibodies bound to lymphocytes from some of the animals. These were the antibodies against CD8 and CD4 antigen, the antibody to C3b-receptor, and the antibody to the HLA-DR antigen. The CD8 antigen-reactive antibody reacted with lymphocytes from mink, cat, dog, and sheep, while the CD4 antigen...

  14. CD8+ Lymphocytes Can Control HIV Infection in vitro by Suppressing Virus Replication

    Science.gov (United States)

    Walker, Christopher M.; Moody, Dewey J.; Stites, Daniel P.; Levy, Jay A.

    1986-12-01

    Lymphocytes bearing the CD8 marker were shown to suppress replication of human immunodeficiency virus (HIV) in peripheral blood mononuclear cells. The effect was dose-dependent and most apparent with autologous lymphocytes; it did not appear to be mediated by a cytotoxic response. This suppression of HIV replication could be demonstrated by the addition of CD8+ cells at the initiation of virus production as well as after several weeks of virus replication by cultured cells. The observations suggest a potential approach to therapy in which autologous CD8 lymphocytes could be administered to individuals to inhibit HIV replication and perhaps progression of disease.

  15. Lymphocytes accelerate epithelial tight junction assembly: role of AMP-activated protein kinase (AMPK.

    Directory of Open Access Journals (Sweden)

    Xiao Xiao Tang

    Full Text Available The tight junctions (TJs, characteristically located at the apicolateral borders of adjacent epithelial cells, are required for the proper formation of epithelial cell polarity as well as for sustaining the mucosal barrier to the external environment. The observation that lymphocytes are recruited by epithelial cells to the sites of infection [1] suggests that they may play a role in the modulation of epithelial barrier function and thus contribute to host defense. To test the ability of lymphocytes to modulate tight junction assembly in epithelial cells, we set up a lymphocyte-epithelial cell co-culture system, in which Madin-Darby canine kidney (MDCK cells, a well-established model cell line for studying epithelial TJ assembly [2], were co-cultured with mouse lymphocytes to mimic an infection state. In a typical calcium switch experiment, the TJ assembly in co-culture was found to be accelerated compared to that in MDCK cells alone. This accelaration was found to be mediated by AMP-activated protein kinase (AMPK. AMPK activation was independent of changes in cellular ATP levels but it was found to be activated by the pro-inflammatory cytokine TNF-alpha. Forced suppression of AMPK, either with a chemical inhibitor or by knockdown, abrogated the accelerating effect of lymphocytes on TJ formation. Similar results were also observed in a co-culture with lymphocytes and Calu-3 human airway epithelial cells, suggesting that the activation of AMPK may be a general mechanism underlying lymphocyte-accelerated TJ assembly in different epithelia. These results suggest that signals from lymphocytes, such as cytokines, facilitate TJ assembly in epithelial cells via the activation of AMPK.

  16. Interference of CD95 expression on human lymphocytes.

    Science.gov (United States)

    Petanova, Jitka; Fucikova, Terezie; Bencko, Vladimir

    2002-02-01

    The study presents the exogenous influence of cadmium in comparison with zinc on the apoptosis of human lymphocytes by CD95 expression and its kinetic changes. The salts of both metals were used in final concentrations of 20 microM in cell cultures with whole blood. The duration of cultivation was 18 and 90 hours. The expression of surface antigens was evaluated by flow cytometry with monoclonal antibodies. In cultures of not stimulated cells we found in average 51.54% CD95 positive lymphocytes. The kinetic study of untreated cells showed elevation after 18 hours of cultivation and a very low expression after 90 hours. The CD95 expression on lymphocytes in cell culture with cadmium and zinc was lower after 18 hours of cultivation than in untreated cells. After 90 hours cultivation we found low levels of CD95 expression on cells treated with cadmium and a great individual variability in the number of positive cells upon the influence of zinc.

  17. WE-A-304-01: Strategies and Technologies for Cranial Radiosurgery Planning: MLC-Based Linac

    Energy Technology Data Exchange (ETDEWEB)

    Kim, G. [University of California, San Diego (United States)

    2015-06-15

    The high fractional doses, stringent requirements for accuracy and precision, and surgical perspective characteristic of intracranial radiosurgery create considerations for treatment planning which are distinct from most other radiotherapy procedures. This session will introduce treatment planning techniques specific to two popular intracranial SRS modalities: Gamma Knife and MLC-based Linac. The basic treatment delivery characteristics of each device will be reviewed with a focus on how those characteristics determine the paradigm used for treatment planning. Basic techniques for treatment planning will be discussed, including considerations such as isodose selection, target and organ-at-risk definition, quality indices, and protection of critical structures. Future directions for SRS treatment planning will also be discussed. Learning Objectives: Introduce the basic physical principles of intracranial radiosurgery and how they are realized in the treatment planning paradigms for Gamma Knife and Linac radiosurgery. Demonstrate basic treatment planning techniques. Discuss metrics for evaluating SRS treatment plan quality. Discuss recent and future advances in SRS treatment planning. D. Schlesinger receives research support from Elekta, AB.

  18. Studies on rabbit lymphocytes in vitro

    Science.gov (United States)

    Sell, S.; Gell, P. G. H.

    1969-01-01

    Anti-allotypic sera that have no known allotypic determinants other than those also present in the genotype of the lymphocyte donor are as able to induce lymphocyte `blast' transformation in vitro as are anti-allotypic sera that do have allotypic determinants that are not present in the lymphocyte donor. Therefore, anti-allotypic sera do not appear to function in the stimulation of blast transformation by providing access for any of the known allotypic determinants into lymphocytes. PMID:5769980

  19. Rapid exacerbation of lymphocytic infundibuloneurohypophysitis

    Science.gov (United States)

    Shibue, Kimitaka; Fujii, Toshihito; Goto, Hisanori; Yamashita, Yui; Sugimura, Yoshihisa; Tanji, Masahiro; Yasoda, Akihiro; Inagaki, Nobuya

    2017-01-01

    Abstract Rationale: Lymphocytic hypophysitis is a relatively rare autoimmune disease defined by lymphocytic infiltration to the pituitary. Its rarity and wide spectrum of clinical manifestations make clarification of the pathology difficult. Here, we describe a case we examined from the primary diagnosis to final discharge, showing the serial progression of lymphocytic infundibuloneurohypophysitis (LINH) to panhypopituitarism with extrapituitary inflammatory invasion in a short period, and responding favorably to high-dose glucocorticoid treatment. Patient concerns: Polyuria, General fatigue and Nausea/Vomiting. Diagnoses: Central diabetes insipidus (CDI), Lymphocytic infundibuloneurohypophysitis (LINH). Interventions: Desmopressin acetate, High-dose glucocorticoid (GC) treatment. Outcomes: He was prescribed desmopressin acetate and subsequently discharged. A month later, he revisited our hospital with general fatigue and nausea/vomiting. A screening test disclosed hypopituitarism with adrenal insufficiency. MRI revealed expanded contrast enhancement to the peripheral extrapituitary lesion. He received high-dose GC treatment and the affected lesion exhibited marked improvement on MRI, along with the recovery of the anterior pituitary function. Lessons: This case demonstrates the potential for classical LINH to develop into panhypopituitarsim. We consider this is the first documentation of approaching the cause of atypical LINH with progressive clinical course from the pathological viewpoint. PMID:28248860

  20. Treatment of chronic lymphocytic leukemia.

    Science.gov (United States)

    Ferrajoli, Alessandra; O'Brien, Susan M

    2004-04-01

    Treatment options for patients with chronic lymphocytic leukemia have changed over the past two decades. This article reviews the experience accumulated with the use of alkylating agents alone and in combination; purine analogues alone and in combination and monoclonal antibodies such as rituximab, and alemtuzumab alone and in combination. The results obtained with different treatment strategies are summarized, compared, and reviewed.

  1. Radiosensitivity of peripheral blood lymphocytes in autoimmune disease

    Energy Technology Data Exchange (ETDEWEB)

    Harris, G. (Kennedy Inst. of Rheumatology, London (UK). Div. of Experimental Pathology); Cramp, W.A.; Edwards, J.C.; George, A.M.; Sabovljev, S.A.; Hart, L.; Hughes, G.R.V. (Hammersmith Hospital, London (UK)); Denman, A.M. (Northwich Park Hospital, Harrow (UK)); Yatvin, M.B. (Wisconsin Clinical Cancer Center, Madison (USA))

    1985-06-01

    The proliferation of peripheral blood lymphocytes, cultured with Con A, can be inhibited by ionizing radiation. Lymphocytes from patients with conditions associated with autoimmunity, such as rheumatoid arthritis, systemic lupus erythematosus and polymyositis, are more radiosensitive than those from healthy volunteers or patients with conditions not associated with autoimmunity. Nuclear material isolated from the lymphocytes of patients with autoimmune diseases is, on average, lighter in density than the nuclear material from most healthy controls. This difference in density is not related to increased sensitivity to ionizing radiation but the degree of post-irradiation change in density (lightening) is proportional to the initial density, i.e. more dense nuclear material always shows a greater upward shift after radiation. The recovery of pre-irradiation density of nuclear material, 1 h after radiation exposure, taken as an indication of DNA repair, correlates with the radiosensitivity of lymphocyte proliferation (Con A response); failure to return to pre-irradiation density being associated with increased sensitivity of proliferative response. These results require extension but, taken with previously reported studied of the effects of DNA methylating agents, support the idea that DNA damage and its defective repair could be important in the aetio-pathogenesis of autoimmune disease.

  2. Fludarabine Phosphate, Radiation Therapy, and Rituximab in Treating Patients Who Are Undergoing Donor Stem Cell Transplant Followed by Rituximab for High-Risk Chronic Lymphocytic Leukemia or Small Lymphocytic Lymphoma

    Science.gov (United States)

    2017-03-27

    Chronic Lymphocytic Leukemia; Prolymphocytic Leukemia; Recurrent Small Lymphocytic Lymphoma; Refractory Chronic Lymphocytic Leukemia; Stage III Chronic Lymphocytic Leukemia; Stage III Small Lymphocytic Lymphoma; Stage IV Chronic Lymphocytic Leukemia; Stage IV Small Lymphocytic Lymphoma; T-Cell Large Granular Lymphocyte Leukemia

  3. Fludarabine Phosphate and Total-Body Irradiation Before Donor Peripheral Blood Stem Cell Transplant in Treating Patients With Chronic Lymphocytic Leukemia or Small Lymphocytic Leukemia

    Science.gov (United States)

    2016-07-18

    B-Cell Prolymphocytic Leukemia; Chronic Lymphocytic Leukemia; Prolymphocytic Leukemia; Recurrent Chronic Lymphocytic Leukemia; Recurrent Small Lymphocytic Lymphoma; Refractory Chronic Lymphocytic Leukemia; T-Cell Prolymphocytic Leukemia

  4. Desensitization oft lymphocyte function by CXCR3 ligands in human hepatocellular carcinoma

    Institute of Scientific and Technical Information of China (English)

    Yu-Qing Liu; Ronnie T. Poon; Jeremy Hughes; Qin-Yu Li; Wan-Ching Yu; Sheung-Tat Fan

    2005-01-01

    AIM: Despite the presence of lymphocyte infiltration, human hepatocellular carcinoma (HCC) is typically a rapidly progressive disease. The mechanism of regulation of lymphocyte migration is poorly understood. In this study,we investigated various factors regulating T cell migration in HCC patients. We examined serum CXC chemokine levels in HCC patients and demonstrated the production of CXC chemokines by HCC cell lines. We determined the effect of both HCC patient serum and tumor cell conditioned supernatant upon lymphocyte expression of chemokine receptor CXCR3 as well as lymphocyte migration. Lastly,we examined the chemotactic responses of lymphocytes derived from HCC patients.METHODS: The serum chemokines IP-10 (CXCL10) and Mig (CXCL9) levels were measured by cytometric bead array (CBA) and the tumor tissue IP-10 concentration was measured by ELISA. The surface expression of CXCR3 on lymphocytes was determined by flow cytometry. The migratory function of lymphocytes to the corresponding chemokines was assessed using an in vitro chemotactic assay. Phosphorylation of extracellular signal-regulated kinase (ERK) was determined by Western blot analysis.RESULTS: Increased levels of IP-10 and Mig were detected in HCC patient serum and culture supernatants of HCC cell lines. The IP-10 concentration in the tumor was significantly higher than that in the non-involved adjacent liver tissues.HCC cell lines secreted functional chemokines that induced a CXCR3-specific chemotactic response of lymphocytes.Furthermore, tumor-cell-derived chemokines induced initial rapid phosphorylation of lymphocyte ERK followed by later inhibition of ERK phosphorylation. The culture of normal lymphocytes with HCC cell line supernatants or medium containing serum from HCC patients resulted in a significant reduction in the proportion of lymphocytes exhibiting surface expression of CXCR3. The reduction in T cell expression of CXCR3 resulted in reduced migration toward the ligand IP-10, and both

  5. Chromosomal aberrations in ovine lymphocytes exposed in vitro to tolylfluanid.

    Science.gov (United States)

    Sutiaková, Irena; Kovalkovičová, Natália; Sutiak, Václav

    2012-01-01

    Chromosomal aberrations have been used as important cytogenetic biomarkers to study the mutagenic effects of different chemicals in vivo and in vitro. Chromosomal aberrations were evaluated in cultures of sheep lymphocytes in vitro exposed to the fungicide tolylfluanid. Lymphocyte cultures from three donors were exposed to four different concentrations of fungicide (1.10(-4) M(.)L; 1.10(-5) M(.)L; 1.10(-6) M(.)L; 1 × 10(-7) M(.)L). Chromosomal analysis showed a significant (P = 0.018 and 0.038 respectively, Anova test, P Tukey test) increase in the frequency of aberrant cells (ABC) in cultures treated with the highest negative experimental concentrations of tolylfluanid (1.10(-4) M(.)L; 1.10(-5) M(.)L) compared to control. Significantly increased numbers of chromatid breaks (7.67 ± 0.58% against 1.67 ± 2.08%, P = 0.009, Anova test, P Tukey test) and chromatid gaps (7.67 ± 1.15% against 2.67 ± 0.58%, P = 0.003, Anova test, P Tukey test) were observed in ovine cultures treated with the highest experimental concentration of tolylfluanid (1.10(-4) M(.)L). Tolylfluanid induced also chromosomal exchanges (P = 0.038, and 0.016 respectively, Anova test, P Tukey test) in ovine cultures treated with the highest experimental concentrations of tolylfluanid (1.10(-4) M(.)L; 1.10(-5) M(.)L). The mitotic index has not shown any statistical differences between the various treatments and control groups. Our results suggest a significant genotoxic effect of tolylfluanid only at the highest concentration in sheep peripheral lymphocytes in vitro.

  6. A genetic algorithm used for the intensity level discretization in MLC leaf sequencing for step and shoot IMRT

    Institute of Scientific and Technical Information of China (English)

    2008-01-01

    The inverse planning for a step-and-shoot plan in intensity-modulated radiotherapy(IMRT)is usually a multiple step process.Before being converted into the MLC segments,the optimum intensity profiles of beams,which are generated by an optimization algorithm,shall be discretized into a few intensity levels.The discretization process of the optimum intensity profiles can induce deviations in the final dose distribution from the original optimum dose distribution.This paper describes a genetic algorithm for the discredzation of given optimum intensity profiles.The algorithm minimizes an objective function written in terms of the intensity levels.Both the dose-based objective function,which is defined by the deviation between the dose distributions before and after the discretization,and the intensity-based objective function,which is defined by the deviation between the optimum intensity profiles and the discretization intensity profiles,have been adopted.To evaluate this algorithm,a series of simulation calculations had been carried out using the present algorithm,the even-spaced discredzation and the k-means clustering algorithm respectively.By comparing the resultant diseretization-induced deviations(DIDs)in intensity profiles and in dose distributions,we have found that the genetic algorithm induced less DIDs in comparison with that induced in the even-spaced discretization or the k-means clustering algorithm.Additionally,it has been found that the DIDs created in the genetic algorithm correlate with the complexity of the intensity profiles that is measured by the"fluence map complexity".

  7. Electrostimulation of rat callus cells and human lymphocytes in vitro

    Energy Technology Data Exchange (ETDEWEB)

    Aro, H.; Eerola, E.; Aho, A.J.; Penttinen, R.

    1984-01-01

    Asymmetrical pulsing low voltage current was supplied via electrodes to cultured rat fracture callus cells and human peripheral blood lymphocytes. The (/sup 3/H)thymidine incorporation of the callus cells and 5-(/sup 125/I)iodo-2'-deoxyuridine incorporation of the lymphocytes were determined. The growth pattern of callus cells (estimated by cellular density) did not respond to electrical stimulation. However, the uptake of (/sup 3/H)thymidine was increased at the early phase of cell proliferation and inhibited at later phases of proliferation. The (/sup 3/H)thymidine uptake of confluent callus cell cultures did not respond to electrical stimulation. Lymphocytes reacted in a similar way; stimulated cells took up more DNA precursor than control cells at the early phase of stimulation. During cell division, induced by the mitogens phytohemagglutinin and Concanavalin-A, the uptake of DNA precursor by stimulated cells was constantly inhibited. The results suggest that electrical stimuli affect the uptake mechanisms of cell membranes. The duality of the effect seems to be dependent on the cell cycle.

  8. Investigating the in vitro effect of taurine on the infant lymphocytes by sister chromatid exchange.

    Science.gov (United States)

    Ergun, Mehmet Ali; Soysal, Yasemin; Kismet, Erol; Akay, Cemal; Dundaroz, Rusen; Ilhan, Mustafan; Imirzalioglu, Necat

    2006-06-01

    Taurine (2-aminoethane sulphonic acid) is normally present in most mammalian tissues and the most abundant free amino acid in lymphocytes. It participates in various important physiological activities including modulation of the functioning of the central nervous system, cell proliferation, viability and prevention of oxidant-induced injury in many tissues. Its levels in human milk are very high which may be the most important difference from cow's milk. In contrast, an inverse association between breast-feeding and carcinogenesis in childhood or later in life has been suggested by several studies. The study group consisted of eight healthy infants. Peripheral blood was collected and lymphocytes were cultured with either Taurine or Mitomycin C (MMC). Sister chromatid exchange in lymphocytes of the infants were calculated. Statistical differences were found between untreated and MMC-treated lymphocytes, untreated and MMC plus taurine-treated lymphocytes, and between MMC and MMC plus taurine-treated lymphocytes (P = 0.012). The results indicated that taurine plays a protective role in MMC-induced sister chromatid exchange in human lymphocytes. The authors suggest that the high levels of taurine found in human milk may induce protecting effects from breast-feeding against DNA damage and malignancy.

  9. TRPML cation channels regulate the specialized lysosomal compartment of vertebrate B-lymphocytes.

    Science.gov (United States)

    Song, Yumei; Dayalu, Rashmi; Matthews, Sharon A; Scharenberg, Andrew M

    2006-12-01

    B-lymphocytes possess a specialized lysosomal compartment, the regulated transformation of which has been implicated in B-cell antigen presentation. Members of the mucolipin (TRPML) family of cation channels have been implicated in regulated vesicular transport in several tissues, but a role for TRPML function in lymphocyte vesicular transport physiology has not been previously described. To address the role of TRPML proteins in lymphocyte vesicular transport, we analyzed the lysosomal compartment in cultured B-lymphocytes engineered to lack TRPML1 or after expression of N- or C-terminal GFP fusion proteins of TRPML1 or TRPML2. Consistent with previous analyses of lymphocytes derived from human patients with mutations in TRPML1, we were not able to detect abnormalities in the lysosomes of TRPML1-deficient DT40 B-lymphocytes. However, while N-terminal GFP fusions of TRPML2 localized to normal appearing lysosomes, C-terminal GFP fusions of either TRPML1 or TRPML2 acted to antagonize endogenous TRPML function, localizing to large vesicular structures, the histological properties of which were indistinguishable from the enlarged lysosomes observed in affected tissues of TRPML1-deficient humans. Endocytosed B-cell receptors were delivered to these enlarged lysosomes, demonstrating that a TRPML-dependent process is required for normal regulation of the specialized lysosome compartment of vertebrate B-lymphocytes.

  10. Automated Scoring and Analysis of Micronucleated Human Lymphocytes.

    Science.gov (United States)

    Callisen, Hannes Heinrich

    Physical and chemical mutagens and carcinogens in our environment produce chromosome abberations in the circulating peripheral blood lymphocytes. The abberations, in turn, give rise to micronuclei when the lymphocytes proliferate in culture. In order to improve the micronucleus assay as a method for screening human populations for chromosome damage, I have (1) developed a high-resolution optical low-light-level micrometry expert system (HOLMES) to digitize and process microscope images of micronuclei in human peripheral blood lymphocytes, (2) defined a protocol of image processing techniques to objectively and uniquely identify and score micronuclei, and (3) analysed digital images of lymphocytes in order to study methods for (a) verifying the identification of suspect micronuclei, (b) classifying proliferating and non-proliferating lymphocytes, and (c) understanding the mechanisms of micronuclei formation and micronuclei fate during cell division. For the purpose of scoring micronuclei, HOLMES promises to (a) improve counting statistics since a greater number of cells can be scored without operator/microscopist fatigue, (b) provide for a more objective and consistent criterion for the identification of micronuclei than the human observer, and (c) yield quantitative information on nuclear and micronuclear characteristics useful in better understanding the micronucleus life cycle. My results on computer aided identification of micronuclei on microscope slides are gratifying. They demonstrate that automation of the micronucleus assay is feasible. Manual verification of HOLMES' results show correct extraction of micronuclei from the scene for 70% of the digitized images and correct identification of the micronuclei for 90% of the extracted objects. Moreover, quantitative analysis on digitized images of lymphocytes using HOLMES has revealed several exciting results: (a) micronuclear DNA content may be estimated from simple area measurements, (b) micronuclei seem to

  11. The toxic effects of melamine on spleen lymphocytes with or without cyanuric acid in mice.

    Science.gov (United States)

    Yin, Rong H; Liu, Jiao; Li, Hua S; Bai, Wen L; Yin, Rong L; Wang, Xin; Wang, Wen C; Liu, Bao S; Han, Xiao H; Han, Jie; He, Jian B; Han, Xiao R

    2014-12-01

    Melamine is an organic nitrogenous compound whose acute toxicity was generally thought to be low in animals. In the present work, we investigated the potential cytotoxic effects of melamine on spleen lymphocytes in mice. In the treated group, morphological changes were observed in cultured lymphocytes in vitro. The co-administration of melamine and cyanuric acid caused a declining tendency in stimulation index of spleen lymphocyte. All treated groups had lower ratios of CD4+/CD8+. Both early apoptotic and late apoptotic/necrotic rates of lymphocyte were significantly higher in the co-administration high groups of melamine and cyanuric acid. Melamine-related toxicity promoted the expression of Bax mRNA, and suppressed the expression of Bcl-2 mRNA in spleen of the treated mice. These results provided useful information for assessing the toxicity of melamine on immune system of mammals, and contributed to the existing toxic profile of melamine.

  12. Studies on poison ivy. In vitro lymphocyte transformation by urushiol-protein conjugates.

    Science.gov (United States)

    Dupuis, G

    1979-12-01

    The isolation and purification of poison ivy urushiol is described. The preparation of urushiol-ski protein and urushiol human serum albumin is also described. Lymphocytes from eleven donor naturally sensitized to poison ivy and from four non-sensitive individuals have been cultured for 5 days in the presence of urushiol-carrier conjugates. Lymphocytes from seven of the eleven sensitive donors responded with a stimulation index greater than 3.0 to urushiol-albumin conjugate. When urushiol-skin protein conjugate was used as a stimulant, lymphocytes from only three of the eleven sensitive donors responded. The results suggest that urushiol-protein conjugates can stimulate sensitive lymphocytes in vitro, although a response is not observed in every individual naturally sensitized to poison ivy.

  13. Molecular In Vivo Imaging Using a Noninvasive Cardiac-Specific MLC-2v Promoter Driven Dual-Gene Recombinant Lentivirus Monitoring System.

    Directory of Open Access Journals (Sweden)

    Miao Zhang

    Full Text Available Our study aimed to demonstrate the feasibility of using the sodium/iodide symporter (NIS to monitor vascular endothelial growth factor (VEGF165 expression in vivo.We constructed a recombinant lentivirus plasmid with the MLC-2v promoter driving the sodium/iodide symporter (NIS reporter gene linked to the VEGF165 gene. Expression of NIS and VEGF gene were identified by Western blot. On days 2 and 54, 99mTc-MIBI imaging was used to evaluate changes in myocardial ischemia. Noninvasive 125I micro-SPECT/CT imaging was used to assess the expression of NIS reporter gene dynamically over the next 2 months.Western blot analysis showed that both NIS and VEGF165 were highly expressed in rat cardiomyoblast H9C2 cells transduced with Lenti-MLC-2v-NIS--VEGF165. 125I micro-SPECT/CT reporter imaging showed higher uptake in mouse myocardium transduced with Lenti-MLC-2v-VEGF165-IRES-NIS. NIS expression peaked on day 1 after transduction followed by a progressive decline to negligible levels by day 21. On day 1, mean 125I activity value in group 1 was higher than that in group 2 (P 0.05. In group 1 (test group, 99mTc-MIBI SPECT/CT revealed improvements in perfusion and wall thickening in the apical anterior wall. Mean IOD values of NIS and CD34 were significantly higher in group 1 than group 3 (P<0.05. Our study proved mean I-125 uptake was significantly correlated with mean IOD value of NIS and CD34 (P<0.05.This study demonstrates the feasibility of using the NIS gene to monitor VEGF165 expression in a mouse myocardial ischemia model.

  14. Metaphase yields from staphylococcal enterotoxin A stimulated peripheral blood lymphocytes of unirradiated and irradiated aged rhesus monkeys

    Science.gov (United States)

    Hill, F. S.; Cox, A. B.; Salmon, Y. L.; Cantu, A. O.; Lucas, J. N.

    1994-01-01

    The mitogen phytohemagglutinin (PHA) works well in both human and cynomolgus monkey (Macaca fascicularis) lymphocyte cultures to stimulate T cell proliferation. T cells from rhesus monkeys (Macaca mulatta) are less responsive than human cells, producing few metaphases when thousands are required, e.g. in biological dosimetry studies. We show that staphylococcal enterotoxin A (SEA), one of the most potent mitogens known, at a concentration of 0.5 microgram/ml stimulated peripheral lymphocytes to grow with a mitotic index (MI) averaging 0.13 metaphases/cell in old, irradiated rhesus macaques. This was significantly greater (p < 0.001) than that produced by PHA (MI < 0.01) in lymphocytes from the same animals. Whole blood was cultured for 96, 120 and 144 h for five irradiated individuals and for two controls. All cells cultured with SEA produced a high MI with a peak response at 120 h whereas the same cultures showed low MI for each PHA stimulated culture.

  15. A virtual photon source model of an Elekta linear accelerator with integrated mini MLC for Monte Carlo based IMRT dose calculation.

    Science.gov (United States)

    Sikora, M; Dohm, O; Alber, M

    2007-08-07

    A dedicated, efficient Monte Carlo (MC) accelerator head model for intensity modulated stereotactic radiosurgery treatment planning is needed to afford a highly accurate simulation of tiny IMRT fields. A virtual source model (VSM) of a mini multi-leaf collimator (MLC) (the Elekta Beam Modulator (EBM)) is presented, allowing efficient generation of particles even for small fields. The VSM of the EBM is based on a previously published virtual photon energy fluence model (VEF) (Fippel et al 2003 Med. Phys. 30 301) commissioned with large field measurements in air and in water. The original commissioning procedure of the VEF, based on large field measurements only, leads to inaccuracies for small fields. In order to improve the VSM, it was necessary to change the VEF model by developing (1) a method to determine the primary photon source diameter, relevant for output factor calculations, (2) a model of the influence of the flattening filter on the secondary photon spectrum and (3) a more realistic primary photon spectrum. The VSM model is used to generate the source phase space data above the mini-MLC. Later the particles are transmitted through the mini-MLC by a passive filter function which significantly speeds up the time of generation of the phase space data after the mini-MLC, used for calculation of the dose distribution in the patient. The improved VSM model was commissioned for 6 and 15 MV beams. The results of MC simulation are in very good agreement with measurements. Less than 2% of local difference between the MC simulation and the diamond detector measurement of the output factors in water was achieved. The X, Y and Z profiles measured in water with an ion chamber (V = 0.125 cm(3)) and a diamond detector were used to validate the models. An overall agreement of 2%/2 mm for high dose regions and 3%/2 mm in low dose regions between measurement and MC simulation for field sizes from 0.8 x 0.8 cm(2) to 16 x 21 cm(2) was achieved. An IMRT plan film verification

  16. SU-E-T-282: Remove Field-Size Limitation Using the Combination of Collimator Rotation and Jaw Tracking with MLC Aperture

    Energy Technology Data Exchange (ETDEWEB)

    Shiu, A; Chiu, R; Cumming, D; Chung, E; Ragab, O; Cui, G [UniversitySouthern California, Los Angeles, CA (United States)

    2014-06-01

    Purpose: this study is to investigate the use of variable-collimators and/or collimator-rotation to overcome the field-size limitation on TrueBeamSTX. Methods: The maximum static field-size is 22- × 40-cm for TrueBeamSTX and 40- × 40-cm for Trilogy. Maximum displacement between adjacent leaf ends at a single carriage position is 15-cm for TBSTX and 14-cm for Trilogy. Our current practice for IMRT treatment at Trilogy is as follows, if MLC leaves travel more than 13.5 cm in the MLC traveling direction, the treatment fields will be split at isocenter to two groups of IMRT fields to cover the large tumors. However, TrueBeamSTX is not limited in this way because the jaws can track each segments of the MLC aperture. Radiation treatment plans for ten patients (different tumor sites) treated at Trilogy with one dimension of the field is greater than 22 cm and the other dimension is less than or equal to 22 cm. All the plans will be converted with jaws tracking each segment's MLC aperture. If necessary, the collimator rotation will also be applied for the planning. Comparison of the converted plans with the original clinical plans will be evaluated and based on the following criterion: (1) tumor coverage; (2) dose sparing to the critical structures; and (3) treatment time. Results: The tumor coverage is compatible between two plans. The dose of critical structures was evaluated in the cumulative DVH for spinal cord, parotids, and brainstem at V20 and V5. In all patients, the maximum reduction in V20 was never more than 5% and was typically less than 1%. In V5, the maximum reduction was 10% and 3% respectively. The significant difference can be found in using the variable collimator to shorten the treatment time. Conclusion: We conclude that we will use the variable-collimator for all IMRT cases on TrueBeamSTX and as a potential backup machine for Trilogy.

  17. Low-Dose Lithium Stabilizes Human Endothelial Barrier by Decreasing MLC Phosphorylation and Universally Augments Cholinergic Vasorelaxation Capacity in a Direct Manner

    Science.gov (United States)

    Bosche, Bert; Molcanyi, Marek; Rej, Soham; Doeppner, Thorsten R.; Obermann, Mark; Müller, Daniel J.; Das, Anupam; Hescheler, Jürgen; Macdonald, R. Loch; Noll, Thomas; Härtel, Frauke V.

    2016-01-01

    Lithium at serum concentrations up to 1 mmol/L has been used in patients suffering from bipolar disorder for decades and has recently been shown to reduce the risk for ischemic stroke in these patients. The risk for stroke and thromboembolism depend not only on cerebral but also on general endothelial function and health; the entire endothelium as an organ is therefore pathophysiologically relevant. Regardless, the knowledge about the direct impact of lithium on endothelial function remains poor. We conducted an experimental study using lithium as pharmacologic pretreatment for murine, porcine and human vascular endothelium. We predominantly investigated endothelial vasorelaxation capacities in addition to human basal and dynamic (thrombin-/PAR-1 receptor agonist-impaired) barrier functioning including myosin light chain (MLC) phosphorylation (MLC-P). Low-dose therapeutic lithium concentrations (0.4 mmol/L) significantly augment the cholinergic endothelium-dependent vasorelaxation capacities of cerebral and thoracic arteries, independently of central and autonomic nerve system influences. Similar concentrations of lithium (0.2–0.4 mmol/L) significantly stabilized the dynamic thrombin-induced and PAR-1 receptor agonist-induced permeability of human endothelium, while even the basal permeability appeared to be stabilized. The lithium-attenuated dynamic permeability was mediated by a reduced endothelial MLC-P known to be followed by a lessening of endothelial cell contraction and paracellular gap formation. The well-known lithium-associated inhibition of inositol monophosphatase/glycogen synthase kinase-3-β signaling-pathways involving intracellular calcium concentrations in neurons seems to similarly occur in endothelial cells, too, but with different down-stream effects such as MLC-P reduction. This is the first study discovering low-dose lithium as a drug directly stabilizing human endothelium and ubiquitously augmenting cholinergic endothelium

  18. Effects of environmental stressors on lymphocyte proliferation in Florida manatees, Trichechus manatus latirostris.

    Science.gov (United States)

    Walsh, Cathy J; Luer, Carl A; Noyes, David R

    2005-02-10

    The health of many Florida manatees (Trichechus manatus latirostris) is adversely affected each year by exposure to cold weather or harmful algal blooms (red tide; Karenia brevis). Exposures can be sublethal, resulting in stressed animals that are rescued and taken to authorized facilities for rehabilitation, or lethal if exposures are prolonged or unusually severe. To investigate whether sublethal environmental exposures can impair immune function in manatees, rendering animals vulnerable to disease or death, mitogen-induced proliferation was assessed in lymphocytes from manatees exposed to cold temperatures (N=20) or red tide (N=19) in the wild, and compared to lymphocyte responses from healthy free-ranging manatees (N=32). All animals sampled for this study were adults. Lymphocytes were stimulated in vitro with either concanavalin A (ConA) or phytohemagglutinin (PHA) and proliferation was assessed after 96 h using incorporation of the thymidine analog, bromodeoxyuridine (BrdU), into newly synthesized DNA. Proliferation of lymphocytes from manatees rescued from exposure to red tide or cold-stress was approximately one-third that of lymphocytes from healthy free-ranging manatees. To examine the direct effects of red tide toxins on lymphocyte function, mitogen-induced proliferation was assessed following co-culture of lymphocytes with K. brevis toxin extracts. Stimulation indices decreased with increasing toxin concentration, with a significant decrease in proliferation occurring in the presence of 400 ng red tide toxins/ml. When lymphocytes from cold-stressed manatees were co-cultured with red tide toxin extracts, proliferative responses were reduced even further, suggesting multiple stressors may have synergistic effects on immune function in manatees.

  19. Opinion: Interactions of innate and adaptive lymphocytes

    Science.gov (United States)

    Gasteiger, Georg; Rudensky, Alexander Y.

    2015-01-01

    Innate lymphocytes, including natural killer (NK) cells and the recently discovered innate lymphoid cells (ILCs) have crucial roles during infection, tissue injury and inflammation. Innate signals regulate the activation and homeostasis of innate lymphocytes. Less well understood is the contribution of the adaptive immune system to the orchestration of innate lymphocyte responses. We review our current understanding of the interactions between adaptive and innate lymphocytes, and propose a model in which adaptive T cells function as antigen-specific sensors for the activation of innate lymphocytes to amplify and instruct local immune responses. We highlight the potential role of regulatory and helper T cells in these processes and discuss major questions in the emerging area of crosstalk between adaptive and innate lymphocytes. PMID:25132095

  20. Protective effect of hawthorn extract against genotoxicity induced by methyl methanesulfonate in human lymphocytes.

    Science.gov (United States)

    Hosseinimehr, Seyed Jalal; Azadbakht, Mohammad; Tanha, Mohammad; Mahmodzadeh, Aziz; Mohammadifar, Sohila

    2011-05-01

    The preventive effect of hawthorn (Crataegus microphylla) fruit extract against genotoxicity induced by methyl methanesulfonate (MMS) has been investigated in human cultured blood lymphocytes. Peripheral blood samples were collected from human volunteers at 0 (10 minutes before), and at 1 and 2 hours after a single oral ingestion of 1 g hawthorn powder extract. At each time point, the whole blood was treated in vitro with MMS (200 µmol) at 24 hours after cell culture, and then the lymphocytes were cultured with mitogenic stimulation to determine the micronuclei in cytokinesis-blocked binucleated cells. The lymphocytes treated with hawthorn and MMS to exhibit a significant decreasing in the incidence of micronucleated binucleated cells, as compared with similarly MMS-treated lymphocytes from blood samples collected at 0 hour. The maximum protection and decreasing in frequency of micronuclei (36%) was observed at 1 hour after ingestion of hawthorn extract. The high performance liquid chromatography (HPLC) analysis showed that hawthorn contained chlorogenic acid, epicatechin and hyperoside. It is obvious that hawthorn, particularly flavonoids constituents with antioxidative activity, reduced the oxidative stress and genotoxicity induced by toxic compounds. This set of data may have an important application for the protection of human lymphocyte from the genetic damage and side effects induced by chemicals hazardous in people.

  1. [Adoptive transfer of T lymphocytes].

    Science.gov (United States)

    Vié, H; Clémenceau, B

    2017-09-01

    Within a few years, the success of treatments based on the use of T-cells armed with a chimeric T-receptor for the CD19 molecule (CAR-T CD19) has revolutionized the perception of adoptive transfer approaches. The levels of responses observed in acute leukemias, of the order of 70-90 % are indeed unprecedented. The medical and financial enthusiasm aroused by these results has led to the current situation where more than 300 clinical trials are under way, against some thirty different antigens. This enthusiasm, well justified by the first successes, must however be tempered by the difficulties associated with the use of these cells. Indeed, the management of patients is made very complex both for medical reasons, because the toxicities associated with these treatments are important, and for technical reasons, because the preparation of T lymphocytes for therapeutic use requires dedicated structures. During this same period, knowledge of the mechanisms of regulation of T lymphocytes and the possibilities offered by synthetic biology and techniques of genome engineering have progressed considerably. Combined, they allow envisaging a true "programming" of the T lymphocytes, intended to improve the efficiency of the treatments and the safety of the patients. Medical and industrial perspectives and the role of these approaches in the arsenal of cancer therapies will depend largely on two conditions: the emergence of a robust demonstration of their effectiveness in solid tumors, and the establishment of an acceptable production and distribution model 1. Copyright © 2017 Elsevier Masson SAS. All rights reserved.

  2. Lymphocyte subpopulations in Sheehan's syndrome.

    Science.gov (United States)

    Atmaca, Hulusi; Araslı, Mehmet; Yazıcı, Zihni Acar; Armutçu, Ferah; Tekin, Ishak Özel

    2013-06-01

    The role of autoimmunity in the development of Sheehan's syndrome is obscure. There are a limited number of studies investigating the immunological alterations accompanying Sheehan's Syndrome. Our objective was to evaluate lymphocyte subsets in these patients. We conducted a cross-sectional clinical study. Cytofluorometry was used for the immunophenotyping of peripheral blood leukocytes from patients with Sheehan's syndrome followed up in the endocrine clinic during 2005-2009. Fifteen consecutive patients (mean age 61.6 ± 11.3, range 34-75 years) and 25 healthy controls (mean age 56.7 ± 10.6, range 34-80 years) were included. There was no statistically significant difference between the groups in terms of mean age. The percentages of CD19(+), CD16(+)/56(+), CD8(+)28(-), γδTCR(+), CD8(+); the total lymphocyte counts; and the ratio of CD8(+)28(-)/CD8(+)28(+) were similar (p > 0.05) between patients and controls. Whereas the leucocyte counts (p = 0.003), the percentage of CD3 (+) DR (+) (p Sheehan's syndrome compared to healthy controls. There was a positive correlation between the duration of illness and the percentage of CD3(+)DR(+) (r = 0.53, p = 0.03) expression. Some peripheral lymphocyte cell subsets show marked variation in patients with Sheehan's syndrome in comparison to matched healthy subjects, which may have implications for altered immune regulation in these patients. High CD3 (+) DR (+) expression that correlates with the duration of illness in Sheehan's patients is suggestive of an ongoing inflammation accompanying the slow progression of pituitary dysfunction in Sheehan's syndrome. It is not clear if these cellular alterations contribute to the cause or consequence of pituitary deficiency in Sheehan's syndrome.

  3. Adipose tissue lymphocytes: types and roles.

    Science.gov (United States)

    Caspar-Bauguil, S; Cousin, B; Bour, S; Casteilla, L; Castiella, L; Penicaud, L; Carpéné, C

    2009-12-01

    Besides adipocytes, specialized in lipid handling and involved in energy balance regulation, white adipose tissue (WAT) is mainly composed of other cell types among which lymphocytes represent a non-negligible proportion. Different types of lymphocytes (B, alphabetaT, gammadeltaT, NK and NKT) have been detected in WAT of rodents or humans, and vary in their relative proportion according to the fat pad anatomical location. The lymphocytes found in intra-abdominal, visceral fat pads seem representative of innate immunity, while those present in subcutaneous fat depots are part of adaptive immunity, at least in mice. Both the number and the activity of the different lymphocyte classes, except B lymphocytes, are modified in obesity. Several of these modifications in the relative proportions of the lymphocyte classes depend on the degree of obesity, or on leptin concentration, or even fat depot anatomical location. Recent studies suggest that alterations of lymphocyte number and composition precede the macrophage increase and the enhanced inflammatory state of WAT found in obesity. Lymphocytes express receptors to adipokines while several proinflammatory chemokines are produced in WAT, rendering intricate crosstalk between fat and immune cells. However, the evidences and controversies available so far are in favour of an involvement of lymphocytes in the control of the number of other cells in WAT, either adipocytes or immune cells and of their secretory and metabolic activities. Therefore, immunotherapy deserves to be considered as a promising approach to treat the endocrino-metabolic disorders associated to excessive fat mass development.

  4. Autoimmune hepatitis in association with lymphocytic colitis.

    LENUS (Irish Health Repository)

    Cronin, Edmond M

    2012-02-03

    Autoimmune hepatitis is a rare, chronic inflammatory disorder which has been associated with a number of other auto-immune conditions. However, there are no reports in the medical literature of an association with microscopic (lymphocytic) colitis. We report the case of a 53-year-old woman with several autoimmune conditions, including lymphocytic colitis, who presented with an acute hepatitis. On the basis of the clinical features, serology, and histopathology, we diagnosed autoimmune hepatitis. To our knowledge, this is the first report of autoimmune hepatitis in association with lymphocytic colitis, and lends support to the theory of an autoimmune etiology for lymphocytic colitis.

  5. Lymphocyte-platelet crosstalk in Graves' disease.

    Science.gov (United States)

    Kuznik, Boris I; Vitkovsky, Yuri A; Gvozdeva, Olga V; Solpov, Alexey V; Magen, Eli

    2014-03-01

    Platelets can modulate lymphocytes' role in the pathophysiology of thyroid autoimmune diseases. The present study was performed to clarify the status of platelet-lymphocyte subpopulations aggregation in circulating blood in patients with Graves' disease (GD). One hundred and fifty patients with GD (GD group) and 45 hyperthyroid patients with toxic multinodular goiter (TMG group) were recruited in the study. Control group consisted 150 healthy subjects. Immunophenotyping of lymphocytes was performed by flow cytometry. Detection of lymphocyte-platelet aggregates (LPAs) was done using light microscope after Ficoll-gradient centrifugation. The group of GD patients exhibited reduced CD8 lymphocyte and higher CD19 cell counts compared with TMG group and healthy controls. A greater number of activated CD3, HLA-DR+ lymphocytes were observed in GD than in TMG group and control group. GD group was characterized by lower blood platelet count (232 ± 89 × 10 cells/µL) than TMG group (251 ± 97 × 10 cells/µL; P < 0.05) and control group (262 ± 95 × 10 cells/µL; P < 0.05). In GD group, more platelet-bound lymphocytes (332 ± 91 /µL) were found than that in TMG group (116 ± 67/µL, P < 0.005) and control group (104 ± 58 /µL; P < 0.001). GD is associated with higher levels of activated lymphocytes and lymphocyte-platelet aggregates.

  6. Vincristine-induced bystander effect in human lymphocytes

    Energy Technology Data Exchange (ETDEWEB)

    Testi, Serena; Azzarà, Alessia; Giovannini, Caterina; Lombardi, Sara [Unità di Genetica, Dipartimento di Biologia, Pisa University, Via Derna 1, 56126 Pisa (Italy); Piaggi, Simona [Dipartimento di Ricerca Traslazionale e delle Nuove Tecnologie in Medicina e Chirurgia, Pisa University, Via Savi 10, 56126 Pisa (Italy); Facioni, Maria Sole [Unità di Genetica, Dipartimento di Biologia, Pisa University, Via Derna 1, 56126 Pisa (Italy); Scarpato, Roberto, E-mail: roberto.scarpato@unipi.it [Unità di Genetica, Dipartimento di Biologia, Pisa University, Via Derna 1, 56126 Pisa (Italy); Research Center of Nutraceuticals and Food for Health, University of Pisa, Pisa (Italy)

    2016-07-15

    Highlights: • We studied whether or not vincristine induced a bystander response in human lymphocytes. • Vincristine significantly increased MN frequencies in mononucleated recipient cells. • ROS or soluble proteins (IL-32 and TGF-β) may account for the observed response. - Abstract: Bystander effect is a known radiobiological effect, widely described using ionizing radiations and which, more recently, has also been related to chemical mutagens. In this study, we aimed to assess whether or not a bystander response can be induced in cultured human peripheral lymphocytes by vincristine, a chemotherapeutic mutagen acting as spindle poison, and by mitomycin-C, an alkylating agent already known to induce this response in human lymphoblastoid cells. Designing a modified ad hoc protocol for the cytokinesis blocked micronucleus (MN) assay, we detected the presence of a dose-dependent bystander response in untreated cultures receiving the conditioned medium (CM) from mitomycin-C (MMC) or vincristine (VCR) treated cultures. In the case of MMC, MN frequencies, expressed as micronucleated binucleates, were: 13.5 ± 1.41 at 6 μM, 22 ± 2.12 at 12 μM or 28.25 ± 5.13 at 15 μM vs. a control value of 4.75 ± 1.59. MN levels for VCR, expressed as micronucleated mononucleates were: 2.75 ± 0.88 at 0.0 μM, 27.25 ± 2.30 at 0.4 μM, 46.25 ± 1.94 at 0.8 μM, 98.25 ± 7.25 at 1.6 μM. To verify that no mutagen residual was transferred to recipient cultures together with the CM, we evaluated MN levels in cultures receiving the medium immediately after three washings following the chemical treatment (unconditioned medium). We further confirmed these results using a cell-mixing approach where untreated lymphocytes were co-cultured with donor cells treated with an effect-inducing dose of MMC or VCR. A distinct production pattern of both reactive oxygen species and soluble mediator proteins by treated cells may account for the differences observed in the manifestation of the

  7. 人FasL基因转染成熟树突状细胞对T淋巴细胞增殖和凋亡的影响%Dendritic Cells Genetically Engineered to Express Fas Ligand Regulate T Lymphocyte Proliferation and Apoptosis

    Institute of Scientific and Technical Information of China (English)

    周浩; 王芳; 王毅; 廖端芳; 陈专华; 邢增术; 王刚; 罗志刚; 秦国庆; 李建军; 钱坤; 蒋鸿涛

    2011-01-01

    To investigate the effects of dendritic cells on T lymphocyte proliferation and apoptosis, providing an in vitro model of clinical transplant immunological tolerance. After mature dendritic cells (mDCs) from peripheral blood of healthy adults was successfully transfected with human FasL gene, mDCs were analyzed for the expression of cell surface molecules, antigen presenting function and their apoptosis. Effects of nDCs on T lymphocyte proliferation and apoptosis were further detected based on co-culture of mDCs and T lymphocytes.The results show that, FasL did not significantly change the expression level of mlC's surface molecules CD40,CD80, CD86 and HLA-DR; FasL did not induce apoptosis of mDC. No effects on the antigen presenting function of mDC were observed as well. The miC transfected with FasL decreased stimulation index and increased apoptosis of allogeneic T-lymphocyte significantly. So that, human mDCs transfected with FasL may regulate T lymphocyte proliferation and apoptosis without alteration of cell surface molecules and antigen presentation characteristics on human mDC.%探讨转染人FasL基因的成熟树突状细胞(DC)对异体T淋巴细胞增殖和凋亡的影响,为实现临床器官移植免疫耐受提供初步实验依据.从健康成年人外周静脉血中获得成熟树突状细胞.将人FasL基因成功转染成熟树突状细胞,检测其表面分子的表达和自身凋亡情况,并对其抗原递呈功能进行分析.从异体健康成人外周血中获取T淋巴细胞,将转染成功的树突状细胞与T淋巴细胞混合培养,检测其对T淋巴细胞增殖和凋亡的影响.结果表明:人FasL基因转染没有明显影响成熟树突状细胞表面分子CD40、CD80、CD86和HLA-DR的表达;没有诱导树突状细胞自身发生凋亡;没有影响DC的抗原递呈功能.转染FasL基因后的树突状细胞使异体T淋巴细胞刺激指数明显下降,凋亡增加.因此认为,人FasL,基因转染对成熟树突状细胞的

  8. In vitro effects of mesenchymal stem cells on secreting function of T lymphocytes and CD4~+CD25~+T cells from patients with immune thrombo-cytopenia

    Institute of Scientific and Technical Information of China (English)

    赵霞

    2014-01-01

    Objective To analyze in vitro the effect of mesenchymal stem cells(MSCs)on secreting cytokines by T lymphocytes and ratio of CD4+CD25+T cells from patients with immune thrombocytopenia(ITP).Methods Human bone marrow-derived MSCs were isolated by Ficoll Hypaque and cultured for proliferating to passage cells.Allogeneic T lymphocytes

  9. Changes of lymphocyte kinetics in the normal rat, induced by the lymphocyte mobilizing agent polymethacrylic acid

    NARCIS (Netherlands)

    Ormai, S.; Hagenbeek, A.; Palkovits, M.; Bekkum, D.W. van

    1973-01-01

    The changes in lymphocyte kinetics induced by the lymphocyte mobilizing agent polymethacrylic acid (PMAA) were studied in the normal rat. Quantitative data are presented concerning the degree of lymphocyte mobilization in the spleen and in various lymph nodes at different times after PMAA administra

  10. Tumor infiltrating lymphocytes in ovarian cancer.

    Science.gov (United States)

    Santoiemma, Phillip P; Powell, Daniel J

    2015-01-01

    The accumulation of tumor infiltrating lymphocytes (TILs) in ovarian cancer is prognostic for increased survival while increases in immunosuppressive regulatory T-cells (Tregs) are associated with poor outcomes. Approaches that bolster tumor-reactive TILs may limit tumor progression. However, identifying tumor-reactive TILs in ovarian cancer has been challenging, though adoptive TIL therapy in patients has been encouraging. Other forms of TIL immunomodulation remain under investigation including Treg depletion, antibody-based checkpoint modification, activation and amplification using dendritic cells, antigen presenting cells or IL-2 cytokine culture, adjuvant cytokine injections, and gene-engineered T-cells. Many approaches to TIL manipulation inhibit ovarian cancer progression in preclinical or clinical studies as monotherapy. Here, we review the impact of TILs in ovarian cancer and attempts to mobilize TILs to halt tumor progression. We conclude that effective TIL therapy for ovarian cancer is at the brink of translation and optimal TIL activity may require combined methodologies to deliver clinically-relevant treatment.

  11. Minocycline Promotes Neurite Outgrowth of PC12 Cells Exposed to Oxygen-Glucose Deprivation and Reoxygenation Through Regulation of MLCP/MLC Signaling Pathways.

    Science.gov (United States)

    Tao, Tao; Feng, Jin-Zhou; Xu, Guang-Hui; Fu, Jie; Li, Xiao-Gang; Qin, Xin-Yue

    2017-04-01

    Minocycline, a semi-synthetic second-generation derivative of tetracycline, has been reported to exert neuroprotective effects both in animal models and in clinic trials of neurological diseases. In the present study, we first investigated the protective effects of minocycline on oxygen-glucose deprivation and reoxygenation-induced impairment of neurite outgrowth and its potential mechanism in the neuronal cell line, PC12 cells. We found that minocycline significantly increased cell viability, promoted neurite outgrowth and enhanced the expression of growth-associated protein-43 (GAP-43) in PC12 cells exposed to oxygen-glucose deprivation/reoxygenation injury. In addition, immunoblots revealed that minocycline reversed the overexpression of phosphorylated myosin light chain (MLC) and the suppression of activated extracellular signal-regulated kinase 1/2 (ERK1/2) caused by oxygen-glucose deprivation/reoxygenation injury. Moreover, the minocycline-induced neurite outgrowth was significantly blocked by Calyculin A (1 nM), an inhibitor of myosin light chain phosphatase (MLCP), but not by an ERK1/2 inhibitor (U0126; 10 μM). These findings suggested that minocycline activated the MLCP/MLC signaling pathway in PC12 cells after oxygen-glucose deprivation/reoxygenation injury, which resulted in the promotion of neurite outgrowth.

  12. Involvement of caveolin-1 in low shear stress-induced breast cancer cell motility and adhesion: Roles of FAK/Src and ROCK/p-MLC pathways.

    Science.gov (United States)

    Xiong, Niya; Li, Shun; Tang, Kai; Bai, Hongxia; Peng, Yueting; Yang, Hong; Wu, Chunhui; Liu, Yiyao

    2017-01-01

    Tumor cells translocating to distant sites are subjected to hemodynamic shear forces during their passage in the blood vessels. Low shear stress (LSS) plays a critical role in the regulation of various aspects of tumor cells functions, including motility and adhesion. Beyond its structural role, caveolin-1 (Cav-1), the important component of caveolae, represents a modulator of several cancer-associated functions as tumor progression and metastasis. However, the role of Cav-1 in regulating tumor cells response to shear stress remains poorly explored. Here, we characterized the role of LSS and Cav-1 in mediating cell motility and adhesion on human breast carcinoma MDA-MB-231 cells. We first showed that LSS exposure promoted cell polarity and focal adhesion (FA) dynamics, thus indicating elevated cell migration. Silencing of Cav-1 leaded to a significantly lower formation of stress fibers. However, LSS exposure was able to rescue it via the alteration of actin-associated proteins expression, including ROCK, p-MLC, cofilin and filamin A. Time-lapse migration assay indicated that Cav-1 expression fostered MDA-MB-231 cells motility and LSS triggered cells to rapidly generate new lamellipodia. Furthermore, Cav-1 and LSS significantly influenced cell adhesion. Taken together, our findings provide insights into mechanisms underlying LSS triggered events mediated by downstream Cav-1, including FAK/Src and ROCK/p-MLC pathways, involved in the reorganization of the cytoskeleton, cell motility, FA dynamics and breast cancer cell adhesion.

  13. Clonal dominance among T-lymphocyte infiltrates in arthritis

    Energy Technology Data Exchange (ETDEWEB)

    Stamenkovic, I.; Stegagno, M.; Wright, K.A.; Krane, S.M.; Amento, E.P.; Colvin, R.B.; Duquesnoy, R.J.; Kurnick, J.T.

    1988-02-01

    Synovial membranes in patients with rheumatoid arthritis as well as other types of chronic destructive inflammatory arthritis contain infiltrates of activated T lymphocytes that probably contribute to the pathogenesis of the disease. In an effort to elucidate the nature of these infiltrates, interleukin 2 (IL-2)-responsive T lymphocytes were grown out of synovial fragments from 14 patients undergoing surgery for advanced destructive inflammatory joint disease. Eleven of the samples examined were from patients with classical rheumatoid arthritis, while three others were obtained from individuals with clinical osteoarthritis. Southern blot analysis of T-cell receptor (TCR) ..beta..-chain genes in 13 of 14 cultures showed distinct rearrangements, indicating that each culture was characterized by the predominance of a limited number of clones. T-cell populations from peripheral blood stimulated with a variety of activators and expanded with IL-2 did not demonstrate evidence of similar clonality in long-term culture. These results suggest that a limited number of activated T-cell clones predominate at the site of tissue injury in rheumatoid synovial membranes as well as in other types of destructive inflammatory joint disease. Further characterization of these T-cell clones may aid our understanding of the pathogenesis of these rheumatic disorders.

  14. Chromosomal instability in the lymphocytes of breast cancer patients

    Directory of Open Access Journals (Sweden)

    Harsimran Kaur

    2009-01-01

    Full Text Available Genomic instability in the tumor tissue has been correlated with tumor progression. In the present study, chromosomal aberrations (CAs in peripheral blood lymphocytes (PBLs of breast tumor patients were studied to assess whether chromosomal instability (CIN in PBLs correlates with aggressiveness of breast tumor (i.e., disease stage and has any prognostic utility. Cultured blood lymphocyte metaphases were scored for aberrations in 31 breast cancer patients and 20 healthy age and sex-matched controls. A variety of CAs, including aneuploidy, polyploidy, terminal deletions, acentric fragments, double minutes, chromatid separations, ring chromosome, marker chromosome, chromatid gaps, and breaks were seen in PBLs of the patients. The CAs in patients were higher than in controls. A comparison of the frequency of metaphases with aberrations by grouping the patients according to the stage of advancement of disease did not reveal any consistent pattern of variation in lymphocytic CIN. Neither was any specific chromosomal abnormality found to be associated with the stage of cancer. This might be indicative of the fact that cancer patients have constitutional CIN, which predisposes them to the disease, and this inherent difference in the level of genomic instability might play a role in disease progression and response to treatment.

  15. Interpretation of NCCN Guideline: Chronic Lymphocytic Leukemia/Small Lymphocytic Lymphoma (Version 1, 2017

    Directory of Open Access Journals (Sweden)

    Lei XIA

    2016-12-01

    Full Text Available Chronic lymphocytic leukemia (CLL is a kind of chronic lymphocyte proliferative disease with corresponding clinical symptoms caused by the accumulation of mature B lymphocytes in peripheral blood, bone marrow and lymphatic tissues. In recent years, great achievements have been reached on the basic research, new prognostic markers, diagnostic criteria and therapeutic methods in CLL. This study mainly interpreted the corresponding diagnosis and treatment of CLL in NCCN Clinical Practice Guidelines in Oncology (NCCN Guidelines®: Chronic Lymphocytic Leukemia/Small Lymphocytic Lymphoma (Version 1, 2017.

  16. Genetics Home Reference: bare lymphocyte syndrome type I

    Science.gov (United States)

    ... Home Health Conditions bare lymphocyte syndrome type I bare lymphocyte syndrome type I Enable Javascript to view ... boxes. Download PDF Open All Close All Description Bare lymphocyte syndrome type I (BLS I) is an ...

  17. The Danish National Chronic Lymphocytic Leukemia Registry

    DEFF Research Database (Denmark)

    da Cunha-Bang, Caspar; Geisler, Christian Hartmann; Enggaard, Lisbeth

    2016-01-01

    AIM: In 2008, the Danish National Chronic Lymphocytic Leukemia Registry was founded within the Danish National Hematology Database. The primary aim of the registry is to assure quality of diagnosis and care of patients with chronic lymphocytic leukemia (CLL) in Denmark. Secondarily, to evaluate...

  18. Reprogramming T cell Lymphocytes to Induced Pluripotent Stem Cells

    Science.gov (United States)

    Bared, Kalia

    The discovery of induced pluripotent stem cells (iPSC) provided a novel technology for the study of development and pharmacology and complement embryonic stem cells (ES) for cell therapy applications. Though iPSC are derived from adult tissue they are comparable to ES cells in their behavior; multi-lineage differentiation and self-renewal. This makes iPSC research appealing because they can be studied in great detail and expanded in culture broadly. Fibroblasts were the first cell type reprogrammed to an iPSC using a retrovirus vector, since then alternative cell types including lymphocytes have been used to generate iPSC. Different types of vectors have also been developed to enhance iPSC formation and quality. However, specific T lymphocyte subsets have not been shown to reprogram to a pluripotent state to date. Here, we proposed to derive iPSC from peripheral blood effector and central memory T cells, reasoning that the resultant iPSC will maintain the epigenetic memory of a T lymphocyte, including the T cell receptor (TCR) gene rearrangement. This epigenetic memory will enable the differentiation and expansion of T cell iPSC into professional T cells containing a specific TCR. These could then be used for cell therapy to target specific antigens, as well as to improve culture techniques to expand T cells in vitro. We studied different gene delivery methods to derive iPSC from different types of T lymphocytes. We assessed the viability of viral transduction using flow cytometry to detect green fluorescent marker contained in the viral construct and quantitative real time polymerase chain reaction (qRT-PCR) to detect Oct4, Klf4, Sox2, and c-Myc gene expression. Our results demonstrate that the Sendai virus construct is the most feasible platform to reprogram T lymphocytes. We anticipate that this platform will provide an efficient and safe approach to derive iPSC from different T cell subsets, including memory T cells.

  19. Inhibitory effect of arctigenin on lymphocyte activation stimulated with PMA/ionomycin.

    Science.gov (United States)

    Sun, Cheng-Hong; Lai, Xin-Qiang; Zhang, Li; Yao, Jing-Chun; Guan, Yong-Xia; Pan, Li-Hong; Yan, Ying

    2014-04-01

    This study investigated the effect of arctigenin (Arc) on the cell activation, cytokines expression, proliferation, and cell-cycle distribution of mouse T lymphocytes. Mouse lymphocytes were prepared from lymph node and treated with Phorbol-12-myristate-13-acetate (PMA)/Ionimycin (Ion) and/or Arc. CD69, CD25, cytokines, proliferation and cell cycle were assayed by flow cytometry. The results showed that, at concentrations of less than 1.00 micromol x L(-1), Arc expressed non-obvious cell damage to cultured lymphocytes, however, it could significantly down-regulate the expression of CD69 and CD25, as well as TNF-alpha, IFN-gamma, IL-2, IL-4, IL-6 and IL-10 on PMA/Ion stimulated lymphocytes. At the same time, Arc could also inhibit the proliferation of PMA/Ion-activated lymphocytes and exhibited lymphocyte G 0/G1 phase cycle arrest. These results suggest that Arc possesses significant anti-inflammatory effects that may be mediated through the regulation of cell activation, cytokines expression and cell proliferation.

  20. 2-Methoxyestradiol induce the conversion of human peripheral blood memory B lymphocytes into plasma cells.

    Science.gov (United States)

    Cayer, Marie-Pierre; Drouin, Mathieu; Proulx, Maryse; Jung, Daniel

    2010-04-15

    2-Methoxyestradiol (2ME), an end-metabolite of 17beta-estradiol, is an antiproliferative agent that is currently being tested in clinical trials for cancer treatment. We hereby report that sub-cytotoxic concentrations of 2ME influence the in vitro proliferation of human peripheral blood B lymphocytes. More surprisingly, we have observed that 2ME induces the conversion of CD138(-) B lymphocytes into CD138(+) cells of phenotype similar to immunoglobulin (Ig)-secreting plasma cells. Normal human B lymphocytes expressing CD138 increased in response to 2ME in a dose-dependent fashion, from 2% at baseline up to 31% in cells cultured in the presence of 0.75 microM 2ME. Moreover, most of the converted cells were also CD27(+) and secreted high levels of IgG (151 microg/10(6)cells/24h). IEF studies revealed that conversion occurred in a polyclonal manner. We then exploited this effect of 2ME to gain further insights into the molecular mechanisms that govern changes in transcription factors involved in plasma cells differentiation. Plasma cells generated by 2ME treatment of normal human B lymphocytes expressed elevated levels of IRF4 and reduced levels of Pax5 and Bcl-6. Similarly, levels of XBP-1 and Blimp-1 transcripts were increased. Our results suggest that the differentiation of peripheral blood B lymphocytes into plasma cells requires a similar modulation of transcription factors expression that for tonsil and bone marrow B lymphocytes.

  1. In vitro effects of plant and mushroom extracts on immunological function of chicken lymphocytes and macrophages

    Science.gov (United States)

    The present study was conducted to examine the effects of milk thistle (Silybum marianum), turmeric (Curcuma longa), reishi mushroom (Ganoderma lucidum), and shiitake mushroom (Lentinus edodes) on innate immunity and tumor cell viability. In vitro culture of chicken spleen lymphocytes with extracts ...

  2. Lymphocyte response to purified Plasmodium falciparum antigens during and after malaria

    DEFF Research Database (Denmark)

    Bygbjerg, I C; Jepsen, S; Theander, T G

    1986-01-01

    The peripheral blood lymphocyte response to affinity purified soluble Plasmodium falciparum antigens from in vitro cultures was studied in seven patients with acute falciparum malaria, on eight occasions, and in 15 persons having had malaria, at various times post infection, on 24 occasions. During...

  3. Intramembrane particles and the organization of lymphocyte membrane proteins.

    Science.gov (United States)

    Kuby, J M; Wofsy, L

    1981-03-01

    An experimental system was developed in which the majority of all lymphocyte cell-surface proteins, regardless of antigenic specificity, could be cross-linked and redistributed in the membrane to determine whether this would induce a corresponding redistribution of intramembrane particles (IMP). Mouse spleen cells were treated with P-diazoniumphenyl- beta-D-lactoside (lac) to modify all exposed cell-surface proteins. Extensive azo- coupling was achieved without significantly reducing cell viability or compromising cellular function in mitogen- or antigen-stimulated cultures. When the lac-modified cell- surface proteins were capped with a sandwich of rabbit antilactoside antibody and fluorescein-goat anti-rabbit Ig, freeze-fracture preparations obtained from these cells revealed no obvious redistribution of IMP on the majority of fracture faces. However, detailed analysis showed a statistically significant 35 percent decrease (P less than 0.01) in average IMP density in the E face of the lac-capped spleen cells compared with control cells, whereas a few E-face micrographs showed intense IMP aggregation. In contrast, there was no significant alteration of P-face IMP densities or distribution. Apparently, the majority of E-face IMP and virtually all P-face IMP densities or distribution. Apparently, the majority of E-face IMP and virtually all P-face IMP do not present accessible antigenic sites on the lymphocyte surface and do not associate in a stable manner with surface protein antigens. This finding suggests that IMP, as observed in freeze-fracture analysis, may not comprise a representative reflection of lymphocyte transmembrane protein molecules and complexes because other evidence establishes: (a) that at least some common lymphocyte surface antigens are indeed exposed portions of transmembrane proteins and (b) that the aggregation of molecules of any surface antigen results in altered organization of contractile proteins at the cytoplasmic face of the membrane.

  4. Loss of telomeric DNA during aging of normal and trisomy 21 human lymphocytes

    Energy Technology Data Exchange (ETDEWEB)

    Vaziri, H.; Uchida, I.; Lan Wei; Harley, C.B. (McMaster Univ., Hamilton, Ontario (Canada)); Schaechter, F.; Cohen, D. (Centre d' Etude du Polymorphisme Humain, Paris (France)); Xiaoming Zhu; Effros, R. (Univ. of California, Los Angeles (United States))

    1993-04-01

    The telomere hypothesis of cellular aging proposes that loss of telomeric DNA (TTAGGG) from human chromosomes may ultimately cause cell-cycle exit during replicative senescence. Since lymphocytes have a limited replicative capacity and since blood cells were previously shown to lose telomeric DNA during aging in vivo, the authors wished to determine (a) whether accelerated telomere loss is associated with the premature immunosenescence of lymphocytes in individuals with Down syndrome (DS) and (b) whether telomeric DNA is also lost during aging of lymphocytes in vitro. To investigate the effects of aging and trisomy 21 on telomere loss in vivo, genomic DNA was isolated from peripheral blood lymphocytes of 140 individuals (age 0--107 years), including 21 DS patients (age 0--45 years). Digestion with restriction enzymes HinfI and RsaI generated terminal restriction fragments (TRFs), which were detected by Southern analysis using a telomere-specific probe ([sup 32]P-(C[sub 3]TA[sub 2])[sub 3]). The rate of telomere loss was calculated from the decrease in mean TRF length, as a function of donor age. DS patients showed a significantly higher rate of telomere loss with donor age (133 [+-] 15 bp/year) compared with age-matched controls (41 [+-] 7.7 bp/year) (P < .0005), suggesting that accelerated telomere loss is a biomarker of premature immunosenescence of DS patients and that it may play a role in this process. Telomere loss during aging in vitro was calculated for lymphocytes from four normal individuals, grown in culture for 10--30 population doublings. The rate of telomere loss was [approximately]120 bp/cell doubling, comparable to that seen in other somatic cells. Moreover, telomere lengths of lymphocytes from centenarians and from older DS patients were similar to those of senescent lymphocytes in culture, which suggests that replicative senescence could partially account for aging of the immune system in DS patients and in elderly individuals. 31 refs., 3 figs.

  5. Active phagocytosis of Mycobacterium tuberculosis (H37Ra) by T lymphocytes (Jurkat cells).

    Science.gov (United States)

    Zhang, Min; Zhu, Qi; Shi, Ming; Liu, Yang; Ma, Lei; Yang, Yining; Feng, Dongyun; Dai, Wen; Zhang, Lin; Kang, Tao; Chen, Ping; He, Ying; Liu, Tingting; Zhao, Qing; Wang, Wenjing; Zhi, Jin; Feng, Guodong; Zhao, Gang

    2015-08-01

    This study aimed to co-culture Jurkat T lymphocytes with inactivated Mycobacterium tuberculosis (Mtb H37Ra), explore whether T lymphocytes could phagocytose H37Ra cells, and determine the underlying mechanism. Jurkat T lymphocytes were co-cultured with H37Ra cells, and confocal laser scanning microscopy, electron microscopy, and flow cytometry techniques were used to identify phagocytosis and elucidate its mechanism. After Jurkat T lymphocytes phagocytosed H37Ra cells, the cell body became larger, with abundant cytoplasm, the portion of the nucleus closest to the bacterium deformed, long and short pseudopodia were extended, and the folds of the cell membrane formed depressions that created phagocytic vesicles surrounding the bacterium. The macropinocytosis inhibitor amiloride and the cytoskeletal inhibitor cytochalasin D were found to inhibit phagocytic efficacy; serum complements might enhance phagocytosis through opsonization. Jurkat T lymphocytes could actively phagocytose inactivated Mtb via the macropinocytotic mechanism. Actin remodeling played an important role in the macropinocytotic process. Serum complements may regulate phagocytosis.

  6. Lymphocytes from wasted mice express enhanced spontaneous and {gamma}-ray-induced apoptosis

    Energy Technology Data Exchange (ETDEWEB)

    Woloschak, G.E. [Argonne National Lab., IL (United States)]|[Loyola Univ. Medical Center, Maywood, IL (United States); Chang-Liu, Chin-Mei [Argonne National Lab., IL (United States); Chung, Jen; Libertin, C.R. [Loyola Univ. Medical Center, Maywood, IL (United States)

    1993-09-01

    Mice bearing the autosomal recessive mutation wasted (wst/wst) display a disease pattern including faulty repair of DNA damage in lymphocytes after radiation exposure, neurologic abnormalities, and immunodeficiency. Many of the features of this mouse model have suggested a premature or increased spontaneous frequency of apoptosis in thymocytes; past work has shown an inability to establish cultured T cell lines, an abnormally high death rate of stimulated T cells in culture, and an increased sensitivity of T cells to the killing effects of ionizing radiations in wst/wst mice relative to controls. The experiments reported here were designed to examine splenic and thymic lymphocytes from wasted and control mice for signs of early apoptosis. Our results revealed enhanced expression of Rp-8 mRNA (associated with apoptosis) in thymic lymphocytes and reduced expression in splenic lymphocytes of wst/wst mice relative to controls; expression of Rp-2 and Td-30 mRNA (induced during apoptosis) were not detectable in spleen or thymus. Higher spontaneous DNA fragmentation was observed in wasted mice than in controls; however, {gamma}-ray-induced DNA fragmentation peaked at a lower dose and occurred to a greater extent in wasted mice relative to controls. These results provide evidence for high spontaneous and {gamma}-ray-induced apoptosis in T cells of wasted mice as a mechanism underlying the observed lymphocyte and DNA repair abnormalities.

  7. Effects of 8-methoxypsoralen (8-MOP) and UVA on human lymphocytes. [Psoriasis

    Energy Technology Data Exchange (ETDEWEB)

    Lischka, G.; Bohnert, E.; Baechtold G.; Jung, E.G.

    1977-09-27

    Peripheral lymphocytes of 37 psoriatic patients are tested before and under PUVA treatment using as parameter the non specific stimulation effect of HgCl2 (10 microgram/ml) in culture, measuring the 3H-thymidine incorporation after the last 16 h of a 5-days culture. Oral 8-MOP in therapeutic doses is decreasing the lymphocyte stimulation as well as 8-MOP together with UVA irradiation during the first week of treatment. After 1 week, the stimulation is, on the contrary, significantly enhanced after irradiation. Lymphocytes isolated by centrifugation over Lymphoprep are submitted to PUVA conditions in petri dishes (Hank's solution 8-MOP 1 microgram/ml, irradiation with 350 nm, 93 to 372 mJ/cm2). The total cell number, the E-rosette formation (as marker for T-lymphocytes) and the EAC-rosette formation (as marker for B-lymphocytes) are determined. PUVA conditions have an energy dependent decreasing effect on the cell number, while the T- and B-cell proportions remain constant. UVA irradiation alone has such an effect only with high energies. 8-MOP without UVA has no significant influence on the cell number.

  8. Lenalidomide and Chronic Lymphocytic Leukemia

    Directory of Open Access Journals (Sweden)

    Ana Pilar González-Rodríguez

    2013-01-01

    Full Text Available Lenalidomide is an oral immunomodulatory drug used in multiple myeloma and myelodysplastic syndrome and most recently it has shown to be effective in the treatment of various lymphoproliferative disorders such as chronic lymphocytic leukemia (CLL and non-Hodgkin lymphoma. The mechanism of action of lenalidomide varies depending on the pathology, and in the case of CLL, it appears to primarily act by restoring the damaged mechanisms of tumour immunosurveillance. This review discusses the potential mechanism of action and efficacy of lenalidomide, alone or in combination, in treatment of CLL and its toxic effects such as tumor lysis syndrome (TLS and tumor flare reaction (TFR, that make its management different from other hematologic malignancies.

  9. Association of Large-Airway Lymphocytic Bronchitis with Bronchiolitis Obliterans Syndrome

    Science.gov (United States)

    Greenland, John R.; Jones, Kirk D.; Hays, Steve R.; Golden, Jeffrey A.; Urisman, Anatoly; Jewell, Nicholas P.; Caughey, George H.

    2013-01-01

    Rationale: Lung transplantation offers great promise for otherwise terminal lung diseases, but the development of bronchiolitis obliterans syndrome (BOS) continues to limit survival. Although acute rejection and lymphocytic bronchiolitis have been identified as risk factors for the development of BOS, it is unclear whether large-airway lymphocytic inflammation conveys the same risk. Objectives: We evaluated lymphocytic bronchitis on endobronchial biopsies as a risk factor for BOS and mortality. Methods: Endobronchial biopsies were collected and graded during surveillance after lung transplantation. We assessed samples with negative cultures collected in the first 90 days from 298 subjects and compared large-airway lymphocytic bronchitis assessed by a 0–2 “E-score” and with standard A and BR pathology scores for acute rejection and small-airway lymphocytic bronchiolitis, respectively. Measurements and Main Results: We found surprisingly little association between large- and small-airway lymphocytic inflammation scores from a given bronchoscopy. Endobronchial lymphocytic bronchitis was more prevalent in subjects in BOS stage 0p and BOS stages 1–3 at the time of biopsy. Within 90 days after transplantation, increasing maximum E-score was associated with greater risk of BOS (adjusted hazard ratio, 1.76; 95% confidence interval, 1.11–2.78; P = 0.02) and in this analysis 90-day maximum E-scores were the only score type predictive of BOS (P < 0.01). Conclusions: These results support a multicenter study to evaluate endoscopic biopsies for the identification of patients at increased risk for BOS. The association of endobronchial lymphocytic inflammation and BOS may have mechanistic implications. PMID:23239157

  10. Signaling in Human and Murine Lymphocytes in Microgravity: Parallels and Contrasts

    Science.gov (United States)

    Neal, Pellis; Alamelu, Sundaresan; Kulkarni, A. D.; Yamauchi, K.

    2006-01-01

    Immune function in space undergoes dramatic changes, some of which are detrimental to lymphocyte function. These changes may lead to significant immune suppression. Studies with human lymphocytes both in space flight and with ground-based models (NASA in vitro ground-based microgravity analog) indicate that T cell activation is inhibited in microgravity. Other lymphocyte functions, such as locomotion, are also inhibited. There is about an 80 percent homology in the immune response of mice to that of humans. A murine model was investigated because of its ability to parallel some microgravity using hind limb suspension. In in vivo antiorthostatically (AOS)-suspended mice, T cell activation is greatly suppressed, with the majority of activation related cytokines being inhibited. PHA activation in lymphocytes derived from AOS mice (in vivo ground-based microgravity analog) is also suppressed. Calcium ionophore studies in human lymphocytes exposed to modeled microgravity indicate that the calcium pathways are probably unaffected in microgravity. IP3 (inositol triphosphate) receptor expression in both human and mouse lymphocytes cultured in modeled microgravity indicate no suppression of calcium signaling. In the human system, microgravity seems to inhibit signaling cascades either at the level of, or up-stream of, Protein Kinase C (PKC). In particular, a membrane event, such as phospholipase C gamma 1 activity in human lymphocytes is affected, with its direct upstream effector, LAT, being deficiently expressed. In the mouse pathway, LAT is undiminished while another critical intermediate, SLP-76, is diminished significantly. This study identifies critical stages in the human and mouse immune systems and in lymphocytes as a function of microgravity.

  11. Damage of chromosoms under irradiation of human blood lymphocytes and development of bystander effect.

    Science.gov (United States)

    Shemetun, O V

    2016-12-01

    the research the distribution of radiation induced damages among chromosomes and their bands in irra diated in vitro human blood lymphocytes and in unirradiated bystander cells.Material and methods of research: cultivation of human peripheral blood lymphocytes by semi micromethod D.A. Hungerford, modeling of radiation induced bystander effect in mixed cultures consisting of irradiated in vitro and non irradiated blood lymphocytes from persons of different gender, GTG staining of metaphase chromosomes and their cytogenetic analysis. Break points in chromosomes under the formation of aberrations were identified in exposed in vitro human peripheral blood lymphocytes in doses 0.25 Gy (95 breaks in 1248 cells) and 1.0 Gy (227 breaks in 726 cells) and in non irradiated bystander cells under their joint cultivation with irradiated in vitro human lymphocytes (51 breaks in 1137 cells at irradiation of adjacent populations of lymphocytes in dose 0.25 Gy and 75 breaks in 1321 cells at irradiation of adjacent population of lymphocytes in a dose 1.0 Gy). The distribution of injuries among the chromo somes and their bands was investigated. in radiation exposed in vitro human peripheral blood lymphocytes as well as in bystander cells the fre quency of damaged bands and number of breaks which localized in them exceeded the control value (p bystander effect, chromosomes were damaged according to their relative length. Location of bands with increasing number of breaks coincided with the «hot spots» of chromosome damage following irradiation and fragile sites. More sensitive to damage were G negative euchromatin chromosome bands, in which were localized 82 88 % breaks. Damageability of telomeric regions in the irradiated cells had no significant difference from the control, while in bystander cells was lower than control value (p < 0.05). O. V. Shemetun.

  12. Molybdate modulates mitogen and cyclosporin responses of human peripheral blood lymphocytes.

    Science.gov (United States)

    Michelis, Fotios V; Delitheos, Andreas; Tiligada, Ekaterini

    2011-07-01

    The trace element molybdenum (Mo) is an essential component of key physiological systems in animals, plants and microorganisms. The molybdate oxoanion MoO(4)(2-) has been demonstrated to cause diverse yet poorly understood biochemical and pharmacological effects, such as non-specific inhibition of phosphatases and stabilization of steroid receptors. This study aimed to investigate the effects of molybdate on the activation of human peripheral blood lymphocytes (hPBLs) ex vivo and its potential interaction with the widely used immunosuppressant drug cyclosporin A (CsA). Lymphocyte activation was evaluated by performing multiple experiments determining blastogenesis in cultured peripheral blood lymphocytes obtained from 5 healthy volunteers, following stimulation induced by phytohemagglutinin (PHA), in the absence or presence of 0.05-10 mM sodium molybdate or/and 2.5-30 μg/mL CsA. Blastogenesis was assessed by a morphometric assay based on the relative proportions of unactivated lymphocytes, activated lymphoblasts and cells with aberrant morphology after PHA-induced activation. Molybdate concentrations up to 1 mM showed no effect on lymphocyte blastogenesis, while higher concentrations exerted immunosuppressive actions on cultured hPBLs. Co-administration of 0.1 mM sodium molybdate with CsA, at doses up to 20 μg/mL, induced no alteration in the response of cultured hPBLs to CsA. However, molybdate potentiated the immunosuppressive action of higher CsA concentrations, implying a likely dose-related synergistic interaction of the two agents in PHA-stimulated blood lymphocytes. These observations are indicative of the possible biological importance of molybdate oxoanions in the modulation of hPBL activation that may have pharmacological consequences during the therapeutic application of immunomodulatory drugs.

  13. Low-dose lithium stabilizes human endothelial barrier by decreasing MLC phosphorylation and universally augments cholinergic vasorelaxation capacity in a direct manner

    Directory of Open Access Journals (Sweden)

    Bert Bosche

    2016-12-01

    Full Text Available Lithium at plasma concentrations up to 1 mmol/L has been used in patients suffering from bipolar disorder for decades and has recently been shown to reduce the risk for ischemic stroke in these patients. The risk for stroke and thromboembolism depend not only on cerebral but also on general endothelial function and health; the entire endothelium as an organ is therefore pathophysiologically relevant. Regardless, the knowledge about the direct impact of lithium on endothelial function remains poor. We conducted an experimental study using lithium as pharmacologic pretreatment for murine, porcine and human vascular endothelium. We predominantly investigated endothelial vasorelaxation capacities in addition to human basal and dynamic (thrombin-/PAR-1 receptor agonist-impaired barrier functioning including myosin light chain phosphorylation (MLC-P. Low-dose therapeutic lithium concentrations (0.4 mmol/L significantly augment the cholinergic endothelium-dependent vasorelaxation capacities of cerebral and thoracic arteries, independently of central and autonomic nerve system influences. Similar concentrations of lithium (0.2-0.4 mmol/L significantly stabilized the dynamic thrombin-induced and PAR-1 receptor agonist-induced permeability of human endothelium, while even the basal permeability appeared to be stabilized. The lithium-attenuated dynamic permeability was mediated by a reduced endothelial MLC-P known to be followed by a lessening of endothelial cell contraction and paracellular gap formation. The well-known lithium-associated inhibition of inositol monophosphatase/glycogen synthase kinase-3-β signaling-pathways involving intracellular calcium concentrations in neurons seems to similarly occur in endothelial cells, too, but with different down-stream effects such as MLC-P reduction. This is the first study discovering low-dose lithium as a drug directly stabilizing human endothelium and ubiquitously augmenting cholinergic endothelium

  14. [Evolution and phylogeny of B lymphocytes].

    Science.gov (United States)

    Claudio-Piedras, Fabiola; Lanz-Mendoza, Humberto

    2016-01-01

    B lymphocytes are one of the most important cell types involved in the immune response of mammals. The origin and evolution of this cellular type is unknown, but the B lymphocyte bona fide appeared first in fish. In this review we analize the principal components of the immune response of invertebrates, their phylogenetic distribution and the permancence of some properties that allowed the emergence of the B lymphocyte. We started from the idea that many of the components that characterize the B lymphocyte are found distributed among the invertebrates, however, it is in the B lymphocyte, where all these components that give this type of cell its identity, converged. The actual knowledge we have in regards of the lymphocytes comes, in the most part, from physiological studies in mammals, being the mice the more representative. The origin of the B lymphocyte, its alternative mechanisms for generating receptor diversity, its immune effector response, and the generation of memory, require an evolutionary and multidisiplinary approach for its study.

  15. 5-HT7 receptors and tryptophan hydroxylase in lymphocytes of rats: mitogen activation, physical restraint or treatment with reserpine.

    Science.gov (United States)

    Urbina, Mary; Arroyo, Rubén; Lima, Lucimey

    2014-01-01

    Serotonin (5-HT)7 receptors in lymphocytes play a relevant role as modulators of T cell functions and might be modified by stress protocols. The aims of this work were to evaluate: (i) the presence of 5-HT7 receptors in specific lymphocyte populations, (ii) the probable modifications of them by inflammatory stress with mitogen and (iii) the effects of physical and pharmacological stress. Blood lymphocytes were isolated by density gradients and differential adhesion to plastic. Concanavalin A (Con A) was systemically administered (500 μg/kg) or added to lymphocyte cultures (2.5 μg/ml, final volume 200 μl). Physical restraint was performed in Plexiglass boxes for 5 h per day for 5 days. Reserpine administration was 2.5 mg/kg for 3 days. Immunocytochemical labeling of CD4+, CD8+ and 5-HT7 receptors, and also tryptophan hydroxylase cells was performed. mRNA of 5-HT7 receptors was evaluated by RT-PCR. Controls were included for each protocol. Con A treatment or culture exposure increased the number of lymphocytes expressing 5-HT7 receptors or tryptophan hydroxylase, as compared to absence of the mitogen. Receptors were present in 12-16% of total rat lymphocytes, in ∼10% of CD4+ and in ∼5% of CD8+ cells from control rats. CD4+ decreased, and CD8+ and 5-HT7 cells increased after physical restraint. Reserpine treatment elevated CD8+ and 5-HT7 cells. Con A and physical restraint, but not reserpine treatment, significantly augmented 5-HT7 receptor mRNA in lymphocytes. Rat lymphocytes, expressing tryptophan hydroxylase, could synthesize 5-HT, functioning as a direct autocrine modulator. The modifications of CD4+, CD8+ and 5-HT7 receptors in lymphocytes by three stress protocols could have an impact on immune responses. In addition, the differential distribution of 5-HT7 receptors indicates potential specific physiopathological roles. © 2014 S. Karger AG, Basel.

  16. Suppressor cell activity in a proliferative disorder of T lymphocytes.

    Science.gov (United States)

    Kupa, A; Thomas, M E; Moore, H; Bradley, J; Zola, H; Hooper, M; Harding, P

    1981-06-01

    We report details of the immunological profile of a patient with the candidiasis endocrinopathy syndrome who has developed T-type chronic lymphocytic leukaemia. The patient is anergic to a panel of delayed hypersensitivity skin tests, and has poor in vitro mitogenic responses, but B cell function in vivo is not impaired. Subsequent functional studies have revealed that cells from the patient have a significant suppressive effect in coculture (P less than 0.05) on the responses of healthy donor lymphocytes (NR) to the mitogen phytohaemagglutinin (PHA). A degree of selectivity for the suppressive effect is suggested by the lack of similar effects on coculture responses to the mitogens concanavalin A (Con A) and pokeweed mitogen (PWM). Mitomycin C treatment of the patient's cells reduced their suppressive activity but significant suppression was still observed in the majority of PHA cocultures. The suppressor activity required the presence of the patient's cells in cocultures, as no suppression was observed when the patient's serum or cell culture supernatant were included instead of the patient's cells in NR cultures.

  17. Metabolism of peripheral lymphocytes, interleukin-2-activated lymphocytes and tumor-infiltrating lymphocytes from sup 31 P NMR studies

    Energy Technology Data Exchange (ETDEWEB)

    Kaplan, O.; Cohen, J.S.; Aebersold, P.

    1989-11-20

    {sup 31}O NMR spectra of tumor-infiltrating lymphocytes (TILs) were found to be significantly diefferent form those of normal peripheral lymphocytes. The greatest difference was in the phosphodiester (PDE) region, mainly in the glycerophosphocholine (GPC) signal. Short-term activation of peripheral lymphocytes with interleukin-2 induced a small increase in ATP levels. In all lumphocytes the phosphomonoester (PME) region is dominated by phosphoethanolamine (PE), while there is an unusual absence of phosphocholine (PC). Perfusion of these cells with high concentrations of choline caused only a minimal increase in PC, indicating that choline kinase is not the rate limiting step of lecithin synthesis in lymphocytes. (author). 13 refs.; 3 figs.; 1 tab.

  18. Folate deficiency and an abnormal lymphocyte deoxyuridine suppression test in monkeys.

    Science.gov (United States)

    Thenen, S W; Hwang, S M; Blocker, D E; Meadows, C A

    1991-01-01

    Cebus albifrons were fed folate-deficient diets in order to assess folate status at the cellular level with the deoxyuridine suppression test. Plasma and red blood cell folates were significantly lower at 2 months, compared to control values. Hematologic signs of megaloblastic anemia occurred after 6 months, with significantly lower hematocrit, hemoglobin and red blood cell number values and increased polymorphonuclear leukocyte lobe counts. Urinary formiminoglutamic acid excretion also was elevated significantly. Whole blood lymphocyte cultures exhibited abnormal deoxyuridine suppression of [3H]-thymidine incorporation into DNA with folate deficiency. Thus this deoxyuridine suppression test can be used in isolated whole blood lymphocytes of these nonhuman primates to document folate deficiency.

  19. Protective effects of b-carotene and silymarin on human lymphocytes

    OpenAIRE

    2012-01-01

    Beta-carotene and silymarin have antioxidant properties against oxidative damage and are used as dietary supplements. The aim of this study was to assess the protective effects of b-carotene and silymarin on healthy human lymphocytes exposed to L-arginine-induced oxidative damage. Study samples were lymphocyte cultures set up from venous blood obtained from 6 healthy individuals (3 males and 3 females). Oxidative DNA damage was induced by L-arginine. b-Carotene and silymarin were added to the...

  20. Activity of vinorelbine on B-chronic lymphocytic leukemia cells in vitro.

    Science.gov (United States)

    Bernabei, P A; Landini, I; Bartolozzi, B; Banchelli, I; Degli Innocenti o Nocentini, A; Santini, V; Ematologia, U O

    1999-01-01

    Vinorelbine (VNR) is a new semi-synthetic Vinca rosea alkaloid that has been employed both in combination and as a single agent, showing a significant antitumour activity. Since little is known about VNR in human leukemia, we studied the in vitro cytotoxic effect of VNR on peripheral blood lymphocytes from 18 patients affected by B-chronic lymphocytic leukemia (CLL), employing the INT assay. VNR inhibited fresh B-CLL cells from 15/18 patients in primary cultures, the ID50 doses ranging from 4 ng/ml to 83 micrograms/ml. These data strongly suggest that VNR could be effective in the treatment of B-CLL.

  1. Study of the positioning of the films of the MLC by a Test of bands and your influence in the clinic dosimetry in IMRT; Estudio del posicionamiento de las laminas del MLC mediante un test de bandas y su influencia en la dosimetria clinica en IMRT

    Energy Technology Data Exchange (ETDEWEB)

    Serrano Zabaleta, S.; Millan Cebrian, E.; Calvo Carrillo, S.; Alba Escorihuela, V.; Garcia Romero, A.; Ortega Pardina, P.; Canella Anoz, M.; Hernandez Vitoria, A.

    2015-07-01

    We performed a test of adjacent bands inspired by the proposed in AAPM Report No. 72, we provides a parameter characterizing the state of the MLC as to the actual position of its blades. This test has been validated by studying repeatability and reproducibility and has found the correlation between the parameter and creep detected by the ILD. Subsequently it has studied the impact of changes in the positioning of the blades on clinical dosimetry in IMRT patients, reconstructing the patient dose by Matrix Evolution team and Compass software, IBA Dosimetry. (Author)

  2. Mycoplasma contamination revisited: mesenchymal stromal cells harboring Mycoplasma hyorhinis potently inhibit lymphocyte proliferation in vitro.

    Directory of Open Access Journals (Sweden)

    Severin Zinöcker

    Full Text Available BACKGROUND: Mesenchymal stromal cells (MSC have important immunomodulatory effects that can be exploited in the clinical setting, e.g. in patients suffering from graft-versus-host disease after allogeneic stem cell transplantation. In an experimental animal model, cultures of rat T lymphocytes were stimulated in vitro either with the mitogen Concanavalin A or with irradiated allogeneic cells in mixed lymphocyte reactions, the latter to simulate allo-immunogenic activation of transplanted T cells in vivo. This study investigated the inhibitory effects of rat bone marrow-derived MSC subsequently found to be infected with a common mycoplasma species (Mycoplasma hyorhinis on T cell activation in vitro and experimental graft-versus-host disease in vivo. PRINCIPAL FINDINGS: We found that M. hyorhinis infection increased the anti-proliferative effect of MSC dramatically, as measured by both radiometric and fluorimetric methods. Inhibition could not be explained solely by the well-known ability of mycoplasmas to degrade tritiated thymidine, but likely was the result of rapid dissemination of M. hyorhinis in the lymphocyte culture. CONCLUSIONS: This study demonstrates the potent inhibitory effect exerted by M. hyorhinis in standard lymphocyte proliferation assays in vitro. MSC are efficient vectors of mycoplasma infection, emphasizing the importance of monitoring cell cultures for contamination.

  3. Suppression of lymphocyte proliferation by marijuana components is related to cell number and cell source

    Energy Technology Data Exchange (ETDEWEB)

    Klein, T.; Pross, S.; Newton, C.; Friedman, H.

    1986-03-05

    Conflicting reports have appeared concerning the effect of marijuana components on immune responsiveness. The authors have observed that the effect of cannabinoids on lymphocyte proliferation varied with both the concentration of the drug and the mitogen used. They now report that at a constant concentration of drug, the cannabinoid effect varied from no effect to suppression depending upon the number of cells in culture and the organ source of the cells. Dispersed cell suspensions of mouse lymph node, spleen, and thymus were prepared and cultured at varying cell numbers with either delta-9-tetrahydrocannabinol or 11-hydroxy-delta-9-tetrahydrocannabinol and various mitogens. Lymphocyte proliferation was analyzed by /sup 3/H-thymidine incorporation. T-lymphocyte mitogen responses in cultures containing high cell numbers were unaffected by the cannabinoids but as cell numbers were reduced a suppression of the response was observed. Furthermore, thymus cells were considerably more susceptible to cannabinoid suppression than cells from either lymph node or spleen. These results suggest that certain lymphocyte subpopulations are more sensitive to cannabinoid suppression and that in addition to drug concentration other variables such as cell number and cell source must be considered when analyzing cannabinoid effects.

  4. Caspase-3-dependent Cell Death in B lymphocyte Caused by Pseudomonas aeruginosa Pyocyanin

    Directory of Open Access Journals (Sweden)

    Heni Susilowati

    2015-08-01

    Full Text Available Objective: The aim of this study was to investigate cellular responses of B lymphocyte to the exposure of pyocyanin and the role of caspase-3 in its molecular mechanism. Methods: B lymphocytes (Raji cells were cultured overnight prior to the experiments. Cell culture in five replications were then exposed to various concentrations of pyocyanin and incubated for 24 h in antibiotics-free medium. MTT assay was performed to analyze the cytotoxicity effect of pyocyanin. In separated experiments, the cells were cultured with pyocyanin and addressed for cell morphological analysis using phase contrast microscope. To study the mechanism involved in pyocyanin-induced cellular damage, immunocytochemical analysis was run for the identification of active caspase-3 protein expression. Results: The results of this study showed that cell viability was decreased in pcyocyanin-treated groups. Statistical analysis using ANOVA (p < 0.05 demonstrated significant different between groups with significant value of 0.000. Pyocyanin induced cell death on B lymphocyte in dose-dependent manner. Nuclear fragmentation was observed in pyocyanin-induced cell death; furthermore, caspase-3 was expressed clearly in cell cytoplasm after 24 h incubation. Conclusion: It is concluded that pyocyanin is capable of inducing cell death on B lymphocyte. Caspase-3 may play important role in the molecular mechanism of pyocyanin-induced cell death.DOI: 10.14693/jdi.v22i2.403

  5. Genotoxic damage in cultured human peripheral blood lymphocytes ...

    African Journals Online (AJOL)

    Falaq Naz

    2012-06-29

    Jun 29, 2012 ... progesterone. They are used to treat various hormonal ... effects among oral contraceptives (OCs) users (females) by using chromosomal aberrations ..... Oral contraception. In: Clinical gynecologic endocrinology and infertility.

  6. A Monte Carlo model for independent dose verification in IMRT and VMAT for the Varian Novalis TX with high definition MLC

    Directory of Open Access Journals (Sweden)

    Luis Vazquez Quino

    2015-09-01

    Full Text Available Purpose: With intensity modulated radiation therapy (IMRT, the physician can prescribe, design and deliver optimized treatment plans that target the tumor and spare adjacent critical structures. The increased conformity of such plans often comes at the expenses of adding significant complexity to the delivery of the treatment. With volumetrically modulated arc therapy (VMAT, in addition to the modulation of the intensity of the radiation beam, other mechanical parameters such as gantry speed and dose rate are varied during treatment delivery. It is therefore imperative that we develop comprehensive and accurate methods to validate such complex delivery techniques prior to the commencement of the patient’s treatment. Methods: In this study, a Monte Carlo simulation was performed for the high definition multileaf collimator (HD-MLC of a Varian Novalis TX linac. Our simulation is based on the MCSIM code and provides a comprehensive model of the linac head. After validating the model in reference geometries, treatment plans for different anatomical sites were simulated and compared against the treatment planning system (TPS dose calculations. All simulations were performed in a cylindrical water phantom as opposed to the patient anatomy, to remove any complexities associated with density effects. Finally, a comparison through gamma analysis of dose plane between the simulation, the TPS and the measurements from the Matrixx array (IBA was conducted to verify the accuracy of our model against both the measurements and the TPS. Results: Gamma analysis of ten IMRT and ten VMAT cases for different anatomical sites was performed, using a 3%/3 mm passing criterion. The average passing rates were 97.5% and 94.3% for the IMRT and the VMAT plans respectively when comparing the MCSIM and TPS dose calculations. Conclusion: In the present work a Monte Carlo model of a Novalis TX linac which has been tested and benchmarked to produce phase-space files for the

  7. The Effects of Lactobacillus acidophilus on the Intestinal Smooth Muscle Contraction through PKC/MLCK/MLC Signaling Pathway in TBI Mouse Model.

    Directory of Open Access Journals (Sweden)

    Bo Sun

    Full Text Available Clinical studies have shown that probiotics influence gastrointestinal motility. However, the molecular mechanisms by which probiotic Lactobacillus modulates intestinal motility in traumatic brain injury (TBI mouse model have not been explored. In the present study, we provided evidence showing that treatment of TBI mice with Lactobacillus acidophilus significantly improved the terminal ileum villus morphology, restored the impaired interstitial cells of Cajal (ICC and the disrupted ICC networks after TBI, and prevented TBI-mediated inhibition of contractile activity in intestinal smooth muscle. Mechanistically, the decreased concentration of MLCK, phospho-MLC20 and phospho-MYPT1 and increased concentration of MLCP and PKC were observed after TBI, and these events mediated by TBI were efficiently prevented by Lactobacillus acidophilus application. These findings may provide a novel mechanistic basis for the application of Lactobacillus acidophilus in the treatment of TBI.

  8. The Effects of Lactobacillus acidophilus on the Intestinal Smooth Muscle Contraction through PKC/MLCK/MLC Signaling Pathway in TBI Mouse Model.

    Science.gov (United States)

    Sun, Bo; Hu, Chen; Fang, Huan; Zhu, Lina; Gao, Ning; Zhu, Jingci

    2015-01-01

    Clinical studies have shown that probiotics influence gastrointestinal motility. However, the molecular mechanisms by which probiotic Lactobacillus modulates intestinal motility in traumatic brain injury (TBI) mouse model have not been explored. In the present study, we provided evidence showing that treatment of TBI mice with Lactobacillus acidophilus significantly improved the terminal ileum villus morphology, restored the impaired interstitial cells of Cajal (ICC) and the disrupted ICC networks after TBI, and prevented TBI-mediated inhibition of contractile activity in intestinal smooth muscle. Mechanistically, the decreased concentration of MLCK, phospho-MLC20 and phospho-MYPT1 and increased concentration of MLCP and PKC were observed after TBI, and these events mediated by TBI were efficiently prevented by Lactobacillus acidophilus application. These findings may provide a novel mechanistic basis for the application of Lactobacillus acidophilus in the treatment of TBI.

  9. Real-time tracking of tumor motions and deformations along the leaf travel direction with the aid of a synchronized dynamic MLC leaf sequencer

    Energy Technology Data Exchange (ETDEWEB)

    Tacke, Martin; Nill, Simeon; Oelfke, Uwe [Deutsches Krebsforschungszentrum (DKFZ), Department of Medical Physics, Im Neuenheimer Feld 280, 69120 Heidelberg (Germany)

    2007-11-21

    Advanced radiotherapeutical techniques like intensity-modulated radiation therapy (IMRT) are based on an accurate knowledge of the location of the radiation target. An accurate dose delivery, therefore, requires a method to account for the inter- and intrafractional target motion and the target deformation occurring during the course of treatment. A method to compensate in real time for changes in the position and shape of the target is the use of a dynamic multileaf collimator (MLC) technique which can be devised to automatically arrange the treatment field according to real-time image information. So far, various approaches proposed for leaf sequencers have had to rely on a priori known target motion data and have aimed to optimize the overall treatment time. Since for a real-time dose delivery the target motion is not known a priori, the velocity range of the leading leaves is restricted by a safety margin to c x v{sub max} while the following leaves can travel with an additional maximum speed to compensate for the respective target movements. Another aspect to be considered is the tongue and groove effect. A uniform radiation field can only be achieved if the leaf movements are synchronized. The method presented in this note is the first to combine a synchronizing sequencer and real-time tracking with a dynamic MLC. The newly developed algorithm is capable of online optimizing the leaf velocities by minimizing the overall treatment time while at the same time it synchronizes the leaf trajectories in order to avoid the tongue and groove effect. The simultaneous synchronization is performed with the help of an online-calculated mid-time leaf trajectory which is common for all leaf pairs and which takes into account the real-time target motion and deformation information. (note)

  10. Vincristine-induced bystander effect in human lymphocytes.

    Science.gov (United States)

    Testi, Serena; Azzarà, Alessia; Giovannini, Caterina; Lombardi, Sara; Piaggi, Simona; Facioni, Maria Sole; Scarpato, Roberto

    2016-07-01

    Bystander effect is a known radiobiological effect, widely described using ionizing radiations and which, more recently, has also been related to chemical mutagens. In this study, we aimed to assess whether or not a bystander response can be induced in cultured human peripheral lymphocytes by vincristine, a chemotherapeutic mutagen acting as spindle poison, and by mitomycin-C, an alkylating agent already known to induce this response in human lymphoblastoid cells. Designing a modified ad hoc protocol for the cytokinesis blocked micronucleus (MN) assay, we detected the presence of a dose-dependent bystander response in untreated cultures receiving the conditioned medium (CM) from mitomycin-C (MMC) or vincristine (VCR) treated cultures. In the case of MMC, MN frequencies, expressed as micronucleated binucleates, were: 13.5±1.41 at 6μM, 22±2.12 at 12μM or 28.25±5.13 at 15μM vs. a control value of 4.75±1.59. MN levels for VCR, expressed as micronucleated mononucleates were: 2.75±0.88 at 0.0μM, 27.25±2.30 at 0.4μM, 46.25±1.94 at 0.8μM, 98.25±7.25 at 1.6μM. To verify that no mutagen residual was transferred to recipient cultures together with the CM, we evaluated MN levels in cultures receiving the medium immediately after three washings following the chemical treatment (unconditioned medium). We further confirmed these results using a cell-mixing approach where untreated lymphocytes were co-cultured with donor cells treated with an effect-inducing dose of MMC or VCR. A distinct production pattern of both reactive oxygen species and soluble mediator proteins by treated cells may account for the differences observed in the manifestation of the bystander effect induced by VCR. In fact, we observed an increased level of ROS, IL-32 and TGF-β in the CM from VCR treated cultures, not present in MMC treated cultures. Copyright © 2016 Elsevier B.V. All rights reserved.

  11. Monoclonal antibodies in chronic lymphocytic leukemia.

    Science.gov (United States)

    Ferrajoli, Alessandra; Faderl, Stefan; Keating, Michael J

    2006-09-01

    Multiple options are now available for the treatment of chronic lymphocytic leukemia. Over the last 10 years, monoclonal antibodies have become an integral part of the management of this disease. Alemtuzumab has received approval for use in patients with fludarabine-refractory chronic lymphocytic leukemia. Rituximab has been investigated extensively in chronic lymphocytic leukemia both as a single agent and in combination with chemotherapy and other monoclonal antibodies. Epratuzumab and lumiliximab are newer monoclonal antibodies in the early phase of clinical development. This article will review the monoclonal antibodies more commonly used to treat chronic lymphocytic leukemia, the results obtained with monoclonal antibodies as single agents and in combination with chemotherapy, and other biological agents and newer compounds undergoing clinical trials.

  12. Chronic lymphocytic leukemia: present status.

    Science.gov (United States)

    Montserrat, E; Rozman, C

    1995-03-01

    Chronic lymphocytic leukemia (CLL) is the form of leukemia which occurs most frequently in Western countries. Its etiology is unknown, and no relationship with viruses or genes has been demonstrated. Epidemiological data suggest that genetic and ambiental factors might be of some significance. Clinical features of CLL are due to the accumulation of leukemic cells in bone marrow and lymphoid organs as well as the immune disturbances that accompany the disease. The prognosis of patients with CLL varies. Treatment is usually indicated by the risk of the individual patient, which is clearly reflected by the stage of the disease. In the early stage (Binet A, Rai O) it is reasonable to defer therapy until disease progression is observed. By contrast, because their median survival is less than five years, patients with more advanced stages require therapy. For almost 50 years, no major advances in the management of CLL, which has revolved around the use of alkylating agents, have been made. In recent years, the therapeutic approach in patients with CLL has changed as a result of the introduction of combination chemotherapy regimens and, in particular, purine analogues. The latter are already the treatment of choice for patients not responding to standard therapies, and their role as front-line therapy is being investigated. Bone marrow transplants are also being increasingly used. It is to be hoped that in years to come the outcome of patients with CLL will be improved by these advances.

  13. Lymphocyte subsets in pediatric migraine.

    Science.gov (United States)

    Cseh, Aron; Farkas, Kristof Mark; Derzbach, Laszlo; Muller, Katalin; Vasarhelyi, Barna; Szalay, Balazs; Treszl, Andras; Farkas, Viktor

    2013-07-01

    Aseptic inflammation due to activated immune cells has been implicated in the pathomechanism of migraine. We measured the prevalence of regulatory T cells (Tregs), along with that of CD4(+)/CD8(+) lymphocytes and their Th1/Th2 commitment in pediatric migraine. Children and adolescents suffering from migraine without aura, migraine with aura and hemiplegic migraine ictally (n = 53, 27, and 20, respectively), also interictally (n = 33) were recruited and compared to 24 healthy children. Our results indicated comparable prevalence of Tregs, CD4(+) and Th1/Th2 committed cells. CD8(+) prevalence was lower, and CD4(+)/CD8(+) ratio was higher in ictal phase irrespective of the subtype of migraine. No association between CD8(+) prevalence and gender, body weight, disease onset and attack duration in migraine subtypes was found. CD8(+) prevalence was normal in patients in interictal phase. These results suggest the absence of major systemic alteration of adaptive immunity in children and adolescents suffering from migraine; however, a transient decrease of CD8(+) prevalence during the ictal phase was detected irrespective of the subtype of migraine.

  14. Obinutuzumab for chronic lymphocytic leukemia.

    Science.gov (United States)

    Rioufol, Catherine; Salles, Gilles

    2014-10-01

    Chronic lymphocytic leukemia (CLL) is a frequent hematological malignancy that is incurable using standard approaches. Two anti-CD20 monoclonal antibodies (mAb), rituximab and ofatumumab, have been approved for CLL treatment. A new glycoengineered type II humanized anti-CD20 mAb, obinutuzumab (GA101), has been developed and demonstrates increased activity against B-cell malignancies by inducing direct cell death and better antibody-dependent cellular cytotoxicity. In a recent randomized Phase III study in patients with newly diagnosed CLL and coexisting conditions, obinutuzumab plus chlorambucil demonstrated significant improvement in progression-free survival and several other outcome parameters, in contrast to rituximab plus chlorambucil. Grade 3-4 infusion-related reactions and neutropenia occurred more frequently in patients who received obinutuzumab compared with those who received rituximab; however, the rate of serious infections was similar. Obinutuzumab represents a promising new option for patients with CLL and must be investigated with other chemotherapy regimens or with new targeted agents.

  15. How T lymphocytes see antigen

    Science.gov (United States)

    Chakraborty, Arup K.

    2009-03-01

    Complex organisms, like humans, have an adaptive immune system that enables us to do battle with diverse pathogens. This flexible system can also go awry, and many diseases are the direct consequence of the adaptive immune system failing to discriminate between markers of self and non-self. The orchestrators of adaptive immunity are a class of cells called T lymphocytes (T cells). T cells recognize minute numbers of molecular signatures of pathogens, and T cell recognition of these molecular markers of non-self is both specific and degenerate. The specific (yet, cross-reactive), diverse, and self-tolerant T cell repertoire is designed in the thymus. I will describe how an approach that brings together theoretical and computational studies (rooted in statistical physics) with experiments (carried out by key collaborators) has allowed us to shed light on the mechanistic principles underlying how T cells respond to pathogens in a digital fashion (``on'' or ``off''), and how this molecular machinery coupled with frustration (a la spin glasses) plays a key role in designing the special properties of the T cell repertoire during development in the thymus.

  16. 一种估算静态MLC调强放疗治疗时间的方法%Estimate the Delivery Time of Static MLC-based IMRT

    Institute of Scientific and Technical Information of China (English)

    张彦新; 戴建荣; 李明辉; 张寅

    2011-01-01

    Objective: To analyze the influence factors of static MLC-based IMRT plans' delivery time and make a simulant formula to estimate it. Methods: Taking Varian 600CD as the object linac and Pinnacle3 (v8.0) as the treatment plan system, the delivery time of static MLC-based IMRT plan was divided into fourparts: beam-on time, MLC moving time, prepare time of original beams and prepare time of additional beams. If we got every part's linear function expression, we got the formula.Each beam's MLC moving time / beam-on time and the ending time could be got from treatment history. The other data of delivery time could be reckoned through these two data. In this study, 30 patients' treatment time data were used to get the coefficients of the formula. Results: The final formula was T = MU/DR+(NBs-NoB-NAB)×2.98+GA/5.78+(NoB-1)×1 7.58+NAB×26.95.The other 6 patients' plan parameters were used to validate this formula. The rates of the calculating values and the real values were 1.03, 0.92, 1.13, 0.99, 0.99, and 1.06. Conclusions: The main factors which influence delivery time of static MLC-based IMRT are beam numbers and segment beam numbers. Reducing beam and segment beam numbers can obvious reduce delivery time on the condition that dose distribution still conform to the clinic and physical criterion.%目的:本研究针对静态多叶准直器(MLC)调强放疗技术,分析影响治疗时间的各种因素,建立数学模型估算治疗时间,为临床评价计划执行效率提供一个工具.方法:以Varian600CD加速器为研究对象,计划采用Philips公司Pinnacle3(v8.0)计划系统设计.将静态MLC调强放疗的治疗时间T分为4个部分:射线照射时间TMU、MLC走位时间TMLC、原射野准备时间TpOB和分野准备时间TPAB.求出各部分的线性函数表达式,即可得到治疗时间的模拟公式.在VARIS治疗记录中可查到两项有关治疗时间的历史记录,一项是每个射野中所有子野的MLC走位和射线照射时长,另一

  17. Bilateral dacryoadenitis complicated by lymphocytic hypophysitis.

    Science.gov (United States)

    Baoke, Hou; Shihui, Wei; Maonian, Zhang; Zhaohui, Li; Zhitong, Zou; Zhigang, Song; Yan, Hei

    2009-09-01

    Three patients developed dacryoadenitis (DA) or lymphocytic pneumonitis before the diagnosis of lymphocytic hypophysitis (LyH). There were two previous reports of concurrence of DA and LyH. Our patients add support to the idea that DA and LyH are manifestations of a systemic autoimmune disease. We suggest that the discovery of DA should prompt imaging and endocrine investigation of LyH.

  18. Expression of tenascin in lymphocytic autoimmune thyroiditis.

    OpenAIRE

    Back, W; Heubner, C; Winter, J.; Bleyl, U

    1997-01-01

    AIMS: To study the distribution of tenascin by immunocytochemistry in autoimmune diseases of the thyroid. METHODS: Thyroids from patients with inflammatory lesions of the thyroid (lymphocytic thyroiditis Hashimoto, Grave's disease, thyroiditis DeQuervain) were studied by immunocytochemistry using antibodies against tenascin, collagen III, and collagen IV. RESULTS: In autoimmune lymphocytic thyroiditis Hashimoto there was a characteristic corona-like staining pattern of tenascin around all act...

  19. Genotoxicity test of self-renovated ceramics in primary human peripheral lymphocytes.

    Science.gov (United States)

    Hua, Nan; Zhu, Huifang; Zhuang, Jing; Chen, Liping

    2014-12-01

    Zirconia-based ceramics is widely used in dentistry. Different compositions of ceramics have different features. Our self-renovated ceramics become more machinable without scarifying its dental restoration properties after adjusting ratio of lanthanum phosphate (LaPO4)/yttrium oxide (Y2O3). In order to evaluate its safety, here, we tested its genotoxicity in primary human peripheral lymphocytes. The human lymphocytes cultured on three groups of different ratios of LaPO4/Y2O3 diphase ceramics for 6 days showed little effect of growth inhibition and similar effect of growth trend to the negative control. Furthermore, single-cell gel electrophoresis (comet assay) indicated that there was no significant difference of the value of tail moment between the tested ceramics and negative control, the IPS Empress II (P > 0.05). Our findings implicate that our self-renovated ceramics do not induce DNA damages in human peripheral lymphocytes and support their future clinic application.

  20. An experimental system for determining the influence of microgravity on B lymphocyte activation and cell fusion

    Science.gov (United States)

    Sammons, D. W.; Zimmermann, U.; Klinman, N. R.; Gessner, P.; Humphreys, R. C.; Emmons, S. P.; Neil, G. A.

    The influence of microgravity on lymphocyte activation is central to the understanding of immunological function in space. Moreover, the adaptation of groundbased technologies to microgravity conditions presents opportunities for biotechnological applications including high efficiency production of antibody forming hybridomas. Because the emerging technology of microgravity hybridoma generation is dependent upon activation and cultivation of B lymphocytes during flight, we have adapted mitogen-driven B lymphocyte stimulation and culture that allows for the in vitro generation of large numbers of antibody forming cells suitable for cell fusion over a period of 1-2 weeks. We believe that this activation and cultivation system can be flown on near-term space flights to test fundamental hypotheses about mammalian cell activation, cell fusion, metabolism, secretion, growth, and bio-separation.

  1. Effects of dental adhesives on micronucleus frequency in peripheral blood lymphocytes in vitro.

    Science.gov (United States)

    Prica, Dunja; Tadin, Antonija; Marović, Danijela; Katunarić, Marina; Prica, Adriana; Galić, Nada

    2013-09-01

    Dental adhesives come into direct contact with oral tissues. Due to this close and long-term contact, the materials should exhibit a high degree of biocompatibility. The aim of this study was to evaluate the genotoxic effect of dental adhesives on human lymphocytes in vitro. Polymerized dental adhesives (Excite, Adper Single Bond 2, Prompt L-pop and OptiBond Solo Plus) were eluted in dimethyl sulfoxide for 1 hour, 24 h and 120 h (5 days). Thereafter, lymphocyte cultures were treated with different concentrations of eluates (0.2 microg/mL, 0.5 microg/mL and 5 microg/mL) obtained from each of the tested materials. Genotoxicity was evaluated by micronucleus test. The chi2-test was used on statistical analysis (p dental adhesives causes genotoxic effects in human lymphocytes. Toxic effect of these dental adhesives increases with the tested material concentration and decreases with the length of elution period.

  2. Lymphocyte migration into syngeneic implanted lymph nodes

    Energy Technology Data Exchange (ETDEWEB)

    Gordeeva, M.S.

    1986-03-01

    To judge the functional activity of lymphocytes of an implanted lymph node (LN), the proliferative response of lymphocytes of the implanted organ in the blast-transformation reaction in vitro and their ability to induce a local graft versus host reaction (GVHR) were determined. The lymphocyte suspension for labeling with /sup 51/Cr was obtained from peripheral LN in different situations from syngeneic mice. The resulting lymphocyte suspension was labeled with a solution of sodium chromate-/sup 51/Cr in a concentration of 20-40 microCi/100.10/sup 6/ cells in 1 ml for 40 min at 37/sup 0/C. The proliferative activity of a suspension of lymphocytes was estimated as incorporation of /sup 3/H-thymidine into DNA during incubation of the cell suspension for 3 days. Data on migration of /sup 51/Cr-labeled cells and the results of the morphological observations revealed marked ability of lymphocytes of the peripheral pool to colonize the regenerating stroma.

  3. Prenatal ontogeny of lymphocyte subpopulations in pigs.

    Science.gov (United States)

    Sinkora, M; Sinkora, J; Reháková, Z; Splíchal, I; Yang, H; Parkhouse, R M; Trebichavsk, I

    1998-12-01

    Although porcine lymphocytes have been classified into numerous subpopulations in postnatal animals, little is known about the ontogeny of these complex cell subsets. Using double- and triple-colour flow cytometry (FCM), we investigated the surface phenotype of fetal lymphoid cells in the thymus, cord blood, spleen and mesenteric lymph nodes at different stages of gestation. It was found that the major lymphocyte subpopulations started to appear at the beginning of the second third of the gestation period, with B cells being the earliest lymphocyte subpopulation to appear in the periphery. The T-cell receptor (TCR) gamma delta+ cells were the earliest detectable T-cell subset, developing first in the thymus and subsequently arriving in the periphery. Later in ontogeny, however, the number of TCRalpha beta+ lymphocytes rapidly increased, becoming the predominant T cells both in the thymus and in the periphery. Cells with the phenotype of adult natural killer cells were also identified in pig fetuses, though their nature and functional roles remain to be investigated. In addition, CD2 was expressed on most B cells whilst very few CD4+ TCRalpha beta+ cells or CD2+ TCRgamma delta+ cells expressed CD8, suggesting that the expression of CD2 and CD8 may reflect the functional status of the cells in postnatal animals. Taken together, this study has provided a systematic analysis of fetal porcine lymphocyte subpopulations and may provide the base for studies to establish the physiological roles of these lymphocyte subsets.

  4. SHARPIN Regulates Uropod Detachment in Migrating Lymphocytes

    Directory of Open Access Journals (Sweden)

    Jeroen Pouwels

    2013-11-01

    Full Text Available SHARPIN-deficient mice display a multiorgan chronic inflammatory phenotype suggestive of altered leukocyte migration. We therefore studied the role of SHARPIN in lymphocyte adhesion, polarization, and migration. We found that SHARPIN localizes to the trailing edges (uropods of both mouse and human chemokine-activated lymphocytes migrating on intercellular adhesion molecule-1 (ICAM-1, which is one of the major endothelial ligands for migrating leukocytes. SHARPIN-deficient cells adhere better to ICAM-1 and show highly elongated tails when migrating. The increased tail lifetime in SHARPIN-deficient lymphocytes decreases the migration velocity. The adhesion, migration, and uropod defects in SHARPIN-deficient lymphocytes were rescued by reintroducing SHARPIN into the cells. Mechanistically, we show that SHARPIN interacts directly with lymphocyte-function-associated antigen-1 (LFA-1, a leukocyte counterreceptor for ICAM-1, and inhibits the expression of intermediate and high-affinity forms of LFA-1. Thus, SHARPIN controls lymphocyte migration by endogenously maintaining LFA-1 inactive to allow adjustable detachment of the uropods in polarized cells.

  5. Entospletinib and Obinutuzumab in Treating Patients With Relapsed Chronic Lymphocytic Leukemia, Small Lymphocytic Lymphoma, or Non-Hodgkin Lymphoma

    Science.gov (United States)

    2017-07-24

    Anemia; B-Cell Prolymphocytic Leukemia; Fatigue; Fever; Grade 1 Follicular Lymphoma; Grade 2 Follicular Lymphoma; Grade 3a Follicular Lymphoma; Hairy Cell Leukemia; Lymphadenopathy; Lymphocytosis; Lymphoplasmacytic Lymphoma; Mantle Cell Lymphoma; Marginal Zone Lymphoma; Night Sweats; Recurrent Chronic Lymphocytic Leukemia; Recurrent Small Lymphocytic Lymphoma; Refractory Chronic Lymphocytic Leukemia; Refractory Small Lymphocytic Lymphoma; Richter Syndrome; Splenomegaly; Thrombocytopenia; Weight Loss

  6. In Vitro Influence of Mycophenolic Acid on Selected Parameters of Stimulated Peripheral Canine Lymphocytes.

    Directory of Open Access Journals (Sweden)

    Maciej Guzera

    Full Text Available Mycophenolic acid (MPA is an active metabolite of mycophenolate mofetil, a new immunosuppressive drug effective in the treatment of canine autoimmune diseases. The impact of MPA on immunity is ambiguous and its influence on the canine immune system is unknown. The aim of the study was to determine markers of changes in stimulated peripheral canine lymphocytes after treatment with MPA in vitro. Twenty nine healthy dogs were studied. Phenotypic and functional analysis of lymphocytes was performed on peripheral blood mononuclear cells cultured with mitogens and different MPA concentrations- 1 μM (10-3 mol/m3, 10 μM or 100 μM. Apoptotic cells were detected by Annexin V and 7-aminoactinomycin D (7-AAD. The expression of antigens (CD3, CD4, CD8, CD21, CD25, forkhead box P3 [FoxP3] and proliferating cell nuclear antigen [PCNA] was assessed with monoclonal antibodies. The proliferation indices were analyzed in carboxyfluorescein diacetate succinimidyl ester (CFSE-labeled cells. All analyses were performed using flow cytometry. The influence of MPA on apoptosis was dependent on the mechanism of cell activation and MPA concentration. MPA caused a decrease in the expression of lymphocyte surface antigens, CD3, CD8 and CD25. Its impact on the expression of CD4 and CD21 was negligible. Its negative influence on the expression of FoxP3 was dependent on cell stimulation. MPA inhibited lymphocyte proliferation. In conclusion, MPA inhibited the activity of stimulated canine lymphocytes by blocking lymphocyte activation and proliferation. The influence of MPA on the development of immune tolerance-expansion of Treg cells and lymphocyte apoptosis-was ambiguous and was dependent on the mechanism of cellular activation. The concentration that MPA reaches in the blood may lead to inhibition of the functions of the canine immune system. The applied panel of markers can be used for evaluation of the effects of immunosuppressive compounds in the dog.

  7. Platelet-Activating Factor Antagonists Decrease Follicular Dendritic-Cell Stimulation of Human B Lymphocytes

    Directory of Open Access Journals (Sweden)

    Halickman Isaac

    2005-06-01

    Full Text Available Abstract Both B-lymphoblastoid cell lines and tonsillar B lymphocytes express receptors for platelet-activating factor (PAF. In lymph node germinal centres, B lymphocytes interact with follicular dendritic cells (FDCs, which present antigen-containing immune complexes to B lymphocytes. FDCs have phenotypic features that are similar to those of stromal cells and monocytes and may therefore be a source of lipid mediators. In this study, we evaluated the effects of the PAF antagonist WEB 2170 on the activation of tonsillar B lymphocytes by FDCs. FDCs were isolated from tonsils by Bovine Serum Albumin (BSA gradient centrifugation. After being cultured for 6 to 10 days, they were incubated with freshly isolated B cells in the presence or absence of the specific PAF receptor antagonist WEB 2170. B-lymphocyte proliferation was assessed by [3H]-thymidine incorporation, and immunoglobulin (Ig G and IgM secretion was assessed by enzyme-linked immunosorbent assay (ELISA. WEB 2170 (10-6 to 10-8 M inhibited [3H]-thymidine incorporation by up to 35% ± 3%. Moreover, the secretion of IgG and IgM was inhibited by up to 50% by WEB 2170 concentrations ranging from 10-6 to 10-8 M. There was no evidence of toxicity by trypan blue staining, and the addition of WEB 2170 to B cells in the absence of FDCs did not inhibit the spontaneous production of IgG or IgM. The effect of the PAF antagonist is primarily on B lymphocytes, as reverse transcription polymerase chain reaction detected little PAF receptor messenger ribonucleic acid (mRNA from FDCs. These data suggest that endogenous production of PAF may be important in the interaction of B lymphocytes with FDCs.

  8. Radioadaptive response in human B- and CD8{sup +} T-lymphocytes as measured by the acridine orange stained micronuclei technique

    Energy Technology Data Exchange (ETDEWEB)

    Kim, H.S.; Choi, J.M.; Yang, K.H.; Kim, C.S.; Lim, Y.K.; Kim, C.S. [Korea Hydro and Nuclear Power Corporation, Radiation Health Research Institute, Seoul (Korea); Woon, J.H. [National Veterinary Research and Quarantine Service, Anyang (Korea)

    2003-07-01

    To investigate (1) the radiosensitive of B- versus T- lymphocytes and (2) the possible application of their sensitivity for adaptive response after treating with an adapting plus a challenge dose. In the present experiments, micronucleus analysis was performed in B- and CD8{sup +} matured T-lymphocytes of eight healthy volunteers exposed to {gamma}-rays. The number of radio-induced micronuclei was significantly higher in B-lymphocytes compared to T-lymphocytes in the dose range from 10 to 100cGy. To investigate adaptive response, whole blood samples were irradiated in vitro with a pretreatment dose of 1cGy {sup 137}Cs {gamma}-irradiation. Six hours after their initiation, groups of cultures were subsequently exposed to a challenge dose of 100cGy {gamma}-irradiation. Following stimulation with PHA and PWM for T- and B-lymphocyte cultivation, lymphocytes were fixed at 72 hours and stained with acridine orange dye. B-lymphocytes exhibited a greater induction of adaptive response than those of CD8{sup +} matured T-lymphocytes, and when pretreated with 1cGy significantly fewer micronuclei induced by the challenge dose of 100cGy {gamma}-irradiation. The results suggest that the lower dose pretreatments are able to induce a significantly higher adaptive response in human B-lymphocytes, and this adaptive response may result from the DNA repair mechanism, which may lead to less residual damage. (author)

  9. Effect of cholinomimetics and adrenomimetics on proliferation of mouse B lymphocytes during primary immune response to protein antigen

    Energy Technology Data Exchange (ETDEWEB)

    Ado, A.D.; Dontsov, V.I.; Gol' dshtein, M.M.

    1985-12-01

    The aim of this investigation was to study the effect of neurotransmitters on proliferation of B lymphocytes induced by specific antigen. Experiments were carried out on female mice. To estimate proliferative activity, lymphocytes enriched with B cells were incubated in medium 199 for 2 h at 37 degrees C in a dose of 2.10/sup 6/-5.10/sup 6/ cells with 2 microCi of /sup 3/H-(methyl)-thymidine. The effect of acetylcholine on incorporation of /sup 3/H-thymidine into B lymphocytes of mice immunized with different doses of antigen during culture is shown. Discordance of effects of adrenalin and acetylcholine on incorporation of /sup 3/H-thymidine into B lymphocytes of mice immunized with different doses of ovalbumin is also shown.

  10. Phenotypic analysis and effects of sequential administration of activated canine lymphocytes on healthy beagles.

    Science.gov (United States)

    Hoshino, Yuki; Takagi, Satoshi; Osaki, Tomohiro; Okumura, Masahiro; Fujinaga, Toru

    2008-06-01

    We attempted to accumulate the basic data for evaluation of activated lymphocyte therapy for small animal medicine. The peripheral blood mononuclear cells (PBMCs) collected from healthy dogs were activated using anti-CD3 antibody and human recombinant (hr) interleukin (IL)-2 and reactivated using hr interferon (IFN)-alpha and hr IL-2. The property of obtained cells was compared with PBMCs. The number of cells was shown to have increased approximately>50 -fold by cultivation. The proportion of CD8+ cells was significantly increased, the cytotoxicity of the cultured cells was revealed to have been reinforced. Additionally, CD56 mRNA levels tended to have increased. The cells obtained by this method were confirmed to be activated lymphocytes. Furthermore, we investigated the effects of sequential administration of the obtained cells to healthy dogs. By sequential administration of the activated lymphocytes, the cell proliferative activity, proportion of CD4+ cells and CD8+ cells, and serum IFN-gamma concentration were shown to have increased, and no severe adverse effects were observed. Consequently, activated lymphocytes could be induced using anti-CD3 antibody and IL-2 in healthy dogs, and sequential administration of activated lymphocytes reinforced the recipient's immunity.

  11. The role of nucleotides in augmentation of lymphocyte locomotion: Adaptional countermeasure development in microgravity analog environments

    Science.gov (United States)

    Sundaresan, Alamelu; Kulkarni, Anil D.; Yamauchi, Keiko; Pellis, Neal R.

    2006-09-01

    Space travel and long-term space residence such as envisaged in the exploration era implicates burdens on the immune system. An optimal immune response is required to countered and with-stand exposure to pathogens. Countermeasure development is an important avenue in space research especially for long-term space exploration. Microgravity exposure causes detrimental effects in lymphocyte functions which may impair immune response. Impaired lymphocyte function can be remedied by bypassing cell membrane events. This is done by using compounds such as Phorbol Myristate Acetate (PMA). Since activation in mouse splenocytes was augmented using nucleotides, it was essential to observe their effects on human lymphocyte locomotion. A nucleotide/nucleoside (NT/NT) mixture from Otsuka Pharmaceuticals (Naruto, Japan) was used at recommended doses. In lymphocytes cultured in modeled microgravity, the NT/NT mixture used orchestrated locomotion recovery by more than 87%, similar to the response documented with PMA in lymphocytes. Both 12µM and 120µM doses worked similarly. These are preliminary results leading to the possible use of the NT/NT mixture to mitigate immune suppression in micro-gravity. More studies in this direction are required to delineate the role of NT/NT on the immune response in microgravity.

  12. Monoclonal Antibody Therapy in Treating Patients With Chronic Lymphocytic Leukemia, Lymphocytic Lymphoma, Acute Lymphoblastic Leukemia, or Acute Myeloid Leukemia

    Science.gov (United States)

    2013-06-03

    Extranodal Marginal Zone B-cell Lymphoma of Mucosa-associated Lymphoid Tissue; Nodal Marginal Zone B-cell Lymphoma; Noncontiguous Stage II Marginal Zone Lymphoma; Noncontiguous Stage II Small Lymphocytic Lymphoma; Recurrent Adult Acute Lymphoblastic Leukemia; Recurrent Adult Acute Myeloid Leukemia; Recurrent Marginal Zone Lymphoma; Recurrent Small Lymphocytic Lymphoma; Refractory Chronic Lymphocytic Leukemia; Splenic Marginal Zone Lymphoma; Stage III Marginal Zone Lymphoma; Stage III Small Lymphocytic Lymphoma; Stage IV Marginal Zone Lymphoma; Stage IV Small Lymphocytic Lymphoma

  13. Construction of HA-1-DC Nucleic-acid Vaccine and Induction of Specific Cytotoxic T Lymphocytes

    Institute of Scientific and Technical Information of China (English)

    WANG Yaya; ZHANG Donghua; HU Jinmei; LIU Wenli; ZHOU hongsheng; ZHANG Lu; LIU Dan; HUANG Zhenqian; TAN Huo

    2007-01-01

    An HA-1-DC nucleic-acid vaccine was constructed to induce anti-leukemia effect after hematopoietic stem cell transplantation (HSCT). DCs were generated from HSCT donors in vitro, and its immunologic activity was assayed by using flow cytometry and mixed lymphocytes reaction.HA-1 gene was electroporated into the cultured DCs to construct a DC nucleic-acid vaccine. After transfection for 48 h, the expression of HA-1 protein could be detected by using Western blot. The DCs were cultured with syngenic lymphocytes to induce specific cytotoxic T lymphocytes (CTLs).The cytoxicity of the CTLs was detected by LDH assay. The results showed that The DCs derived from peripheral blood monocytes (PBMCs) expressed the phenotype of DCs, and were effective in stimulating proliferation of the allogenic lymphocytes. After electroporating for 48-h, HA-1 protein was detected by using Western blot. The cytotoxity of inducing CTLs was higher than the control group. It was suggested that minor histocompatibility antigen HA-1 could be considered as a target of immunotherapy against leukemia after HSCT.

  14. Characterization of the atypical lymphocytes in African swine fever

    Directory of Open Access Journals (Sweden)

    Z. A. Karalyan

    2016-07-01

    Full Text Available Aim: Atypical lymphocytes usually described as lymphocytes with altered shape, increased DNA amount, and larger size. For analysis of cause of genesis and source of atypical lymphocytes during African swine fever virus (ASFV infection, bone marrow, peripheral blood, and in vitro model were investigated. Materials and Methods: Atypical lymphocytes under the influence of ASFV were studied for morphologic, cytophotometric, and membrane surface marker characteristics and were used in vivo and in vitro models. Results: This study indicated the increased size, high metabolic activity, and the presence of additional DNA amount in atypical lymphocytes caused by ASFV infection. Furthermore, in atypical lymphocytes, nuclear-cytoplasmic ratio usually decreased, compared to normal lymphocytes. In morphology, they looking like lymphocytes transformed into blasts by exposure to mitogens or antigens in vitro. They vary in morphologic detail, but most of them are CD2 positive. Conclusions: Our data suggest that atypical lymphocytes may represent an unusual and specific cellular response to ASFV infection.

  15. Characterization of the atypical lymphocytes in African swine fever

    Science.gov (United States)

    Karalyan, Z. A.; Ter-Pogossyan, Z. R.; Abroyan, L. O.; Hakobyan, L. H.; Avetisyan, A. S.; Karalyan, N. Yu; Karalova, E. M.

    2016-01-01

    Aim: Atypical lymphocytes usually described as lymphocytes with altered shape, increased DNA amount, and larger size. For analysis of cause of genesis and source of atypical lymphocytes during African swine fever virus (ASFV) infection, bone marrow, peripheral blood, and in vitro model were investigated. Materials and Methods: Atypical lymphocytes under the influence of ASFV were studied for morphologic, cytophotometric, and membrane surface marker characteristics and were used in vivo and in vitro models. Results: This study indicated the increased size, high metabolic activity, and the presence of additional DNA amount in atypical lymphocytes caused by ASFV infection. Furthermore, in atypical lymphocytes, nuclear-cytoplasmic ratio usually decreased, compared to normal lymphocytes. In morphology, they looking like lymphocytes transformed into blasts by exposure to mitogens or antigens in vitro. They vary in morphologic detail, but most of them are CD2 positive. Conclusions: Our data suggest that atypical lymphocytes may represent an unusual and specific cellular response to ASFV infection. PMID:27536044

  16. Flow cytometric analysis of lymphocytes and lymphocyte subpopulations in induced sputum from patients with asthma

    Directory of Open Access Journals (Sweden)

    Yutaro Shiota

    2000-01-01

    Full Text Available Study objectives were to compare the numbers of lymphocytes and lymphocyte subpopulations in induced sputum from asthmatic patients and from healthy subjects, and to determine the effect of inhaled anti-asthmatic steroid therapy on these cell numbers. Hypertonic saline inhalation was used to non-invasively induce sputum samples in 34 patients with bronchial asthma and 21 healthy subjects. The sputum samples were reduced with dithioerythritol and absolute numbers of lymphocytes and lymphocyte subpopulations were assessed by direct immunofluorescence and flow cytometry. To assess the effect of beclomethasone dipropionate (BDP on induced sputum, numbers of lymphocytes and lymphocyte subpopulations in sputum also were evaluated after 4 weeks of BDP inhalation treatment in seven asthmatic patients. An adequate sample was obtained in 85.3% of patients with asthma and in 79.2% of the healthy subjects. Induced sputum from patients with asthma had increased numbers of lymphocytes (P = 0.009; CD4+ cells (P = 0.044; CD4+ cells-bearing interleukin-2 receptor (CD25; P = 0.016; and CD4+ cells bearing human histocompatibility leukocyte antigen (HLA-DR (P = 0.033. CD8+ cells were not increased in asthmatic patients. In patients treated with inhaled steroids, numbers of lymphocytes, CD4+ cells, CD25-bearing CD4+ cells and HLA-DR-bearing CD4+ cells in sputum decreased from pretreatment numbers (P = 0.016, 0.002, 0.003 and 0.002, respectively. Analysis of lymphocytes in induced sputum by flow cytometry is useful in assessing bronchial inflammation, and activated CD4+ lymphocytes may play a key role in the pathogenesis of airway inflammation in bronchial asthma.

  17. FcgammaRIIb expression on human germinal center B lymphocytes.

    Science.gov (United States)

    Macardle, Peter J; Mardell, Carolyn; Bailey, Sheree; Wheatland, Loretta; Ho, Alice; Jessup, Claire; Roberton, Donal M; Zola, Heddy

    2002-12-01

    IgG antibody can specifically suppress the antibody response to antigen. This has been explained by the hypothesis that signaling through the B cell antigen receptor is negatively modulated by the co-ligation of immunoglobulin with the receptor for IgG, FcgammaRIIb. We hypothesized that inhibitory signaling through FcgammaRIIb would be counter-productive in germinal center cells undergoing selection by affinity maturation, since these cells are thought to receive a survival/proliferative signal by interacting with antigen displayed on follicular dendritic cells. We have identified and characterized a population of B lymphocytes with low/negative FcgammaRIIb expression that are present in human tonsil. Phenotypically these cells correspond to germinal center B cells and comprise both centroblast and centrocyte populations. In examining expression at the molecular level we determined that these B cells do not express detectable mRNA for FcgammaRIIb. We examined several culture conditions to induce expression of FcgammaRIIb on germinal center cells but could not determine conditions that altered expression. We then examined the functional consequence of cross-linking membrane immunoglobulin and the receptor for IgG on human B lymphocytes. Our results cast some doubt on the value of anti-IgG as a model for antigen-antibody complexes in studying human B cell regulation.

  18. [Radiation-induced "bystander effect" revealed by means of adaptive response in cocultured lymphocytes from humans of different genders].

    Science.gov (United States)

    Kolesnikova, I S; Vorobtsova, I E

    2011-01-01

    The "bystander effect" was investigated in mixed cultures of lymphocytes from humans of opposite genders. Development of the adaptive response (AR) in non-irradiated female/male cells was estimated after adaptive pretreatment of opposite gender lymphocytes, chromosome aberrations being evaluated. Experiments were performed using two schedules of adaptive (0.05 Gy) and challenging (1 Gy) irradiations: G0-G1 and G1-G1. The results obtained indicate the development of a mediated adaptive response ("bystander effect") in the lymphocytes neighboring pre-irradiated cells, as well as the influence of a time scheme of adapting and challenging irradiations on the amount of induced chromosome aberrations in mixed cultures and a possible dependence of the adaptive response intensity on the donor gender.

  19. Nodular lymphocyte-predominant Hodgkin lymphoma.

    Science.gov (United States)

    Savage, Kerry J; Mottok, Anja; Fanale, Michelle

    2016-07-01

    Nodular lymphocyte-predominant Hodgkin lymphoma (NLPHL) is a rare subtype of Hodgkin lymphoma with distinct clinicopathologic features. It is typified by the presence of lymphocyte predominant (LP) cells, which are CD20(+) but CD15(-) and CD30(-) and are found scattered amongst small B lymphocytes arranged in a nodular pattern. Despite frequent and often late or multiple relapses, the prognosis of NLPHL is very favorable. There is an inherent risk of secondary aggressive non-Hodgkin lymphoma (NHL) and studies support that risk is highest in those with splenic involvement at presentation. Given disease rarity, the optimal management is unclear and opinions differ as to whether treatment paradigms should be similar to or differ from those for classical Hodgkin lymphoma (CHL). This review provides an overview of the existing literature describing pathological subtypes, outcome and treatment approaches for NLPHL.

  20. T cell immunity using transgenic B lymphocytes

    Science.gov (United States)

    Gerloni, Mara; Rizzi, Marta; Castiglioni, Paola; Zanetti, Maurizio

    2004-03-01

    Adaptive immunity exists in all vertebrates and plays a defense role against microbial pathogens and tumors. T cell responses begin when precursor T cells recognize antigen on specialized antigen-presenting cells and differentiate into effector cells. Currently, dendritic cells are considered the only cells capable of stimulating T lymphocytes. Here, we show that mature naïve B lymphocytes can be genetically programmed by using nonviral DNA and turned into powerful antigen-presenting cells with a dual capacity of synthesis and presentation of antigen to T cells in vivo. A single i.v. injection of transgenic lymphocytes activates T cell responses reproducibly and specifically even at very low cell doses (102). We also demonstrate that T cell priming can occur in the absence of dendritic cells and results in immunological memory with protective effector functions. These findings disclose aspects in the regulation of adaptive immunity and indicate possibilities for vaccination against viruses and cancer in humans.

  1. MiR-31 Regulates Rho-Associated Kinase-Myosin Light Chain (ROCK-MLC) Pathway and Inhibits Gastric Cancer Invasion: Roles of RhoA.

    Science.gov (United States)

    Chen, Zhuo; Liu, Shengnan; Xia, Yuan; Wu, Kejian

    2016-12-01

    BACKGROUND This study evaluated how the expression of miR-31 can be used to detect gastric cancer (GC) to help illuminate the role of miR-31 and RhoA in GC cells. MATERIAL AND METHODS We carried out our experiments using tissue specimens from 70 GC patients. The relative expression of miR-31 and RhoA mRNA in tissues and cells was detected by RT-PCR. The expression level of RhoA protein was detected by immunohistochemistry. GC cell line BGC-823 was transfected with six groups of vectors: blank group, NC (negative control) group, miR-31 mimics group, miR-31 mimics + RhoA group, miR-31 mimics + ROCK group, and miR-31 mimics + MLCK agonist group. AGS cells were also transfected with six groups of vectors: blank group, NC group, miR-31 inhibitor group, miR-31 inhibitor + RhoA siRNA group, miR-31 inhibitor + ROCK siRNA group, and miR-31 inhibitor + MLCK inhibitor group. Transwell assay was performed to detect the invasion and migration of cells. The protein expression in different transfected groups was detected using Western blotting. RESULTS GC tissues exhibited significantly lower levels of miR-31 expression compared to pericarcinous tissues (p<0.01). Moreover, a significantly higher expression of RhoA in GC tissues was observed (p<0.05). MiR-31 inhibited RhoA expression by binding to 3'UTR of mRNA, whereas miR-31 mimics significantly decreased the number of invaded and migrated cells (p<0.05). The activation of RhoA, ROCK, and phosphorylation of MLC remarkably exacerbate the invasion and migration ability of GC cells (p<0.05). CONCLUSIONS We found miR-31 could downregulate the ROCK/MLC pathway by inhibiting the expression of RhoA in order to suppress the invasion and migration of GC cells.

  2. An intrinsic GABAergic system in human lymphocytes.

    Science.gov (United States)

    Dionisio, Leonardo; José De Rosa, María; Bouzat, Cecilia; Esandi, María Del Carmen

    2011-01-01

    γ-amino butyric acid (GABA) is an ubiquitous neurotransmitter in the central nervous system and it is also present in non-neuronal cells. In this study we investigated the presence of neuronal components of the GABAergic system in lymphocytes and its functional significance. By using RT-PCR we detected mRNA expression of different components of the GABAergic system in resting and mitogen-activated lymphocytes: i) GAD67, an isoform of the enzyme that synthetizes GABA; ii) VIAAT, the vesicular protein involved in GABA storage; iii) GABA transporters (GAT-1 and GAT-2); iv) GABA-T, the enzyme that catabolizes GABA; and v) subunits that conform ionotropic GABA receptors. The presence of VIAAT protein in resting and activated cells was confirmed by immunocytochemistry. The functionality of GABA transporters was evaluated by measuring the uptake of radioactive GABA. The results show that [(3)H]GABA uptake is 5-fold higher in activated than in resting lymphocytes. To determine if GABA subunits assemble into functional channels, we performed whole-cell recordings in activated lymphocytes. GABA and muscimol, a specific agonist of ionotropic GABA receptors, elicit macroscopic currents in about 10-15% of the cells. Finally, by using [(3)H]thymidine incorporation assays, we determined that the presence of agonists of GABA receptor during activation inhibits lymphocyte proliferation. Our results reveal that lymphocytes have a functional GABAergic system, similar to the neuronal one, which may operate as a modulator of T-cell activation. Pharmacological modulation of this system may provide new approaches for regulation of T-cell response. Copyright © 2010 Elsevier Ltd. All rights reserved.

  3. Two small lymphocyte subpopulations in human peripheral blood. I. Purification and surface marker profiles

    DEFF Research Database (Denmark)

    Hokland, M; Hokland, P; Heron, I

    1978-01-01

    By means of simple rosette sedimentation methods two subsets from human peripheral blood lymphocytes have been isolated: (1) (E, Fc)- and (2) (E, Ig)-. The first subset was obtained by centrifuging suspensions of macrophage-depleted PBL in which E and EA rosettes had been allowed to form simultan......By means of simple rosette sedimentation methods two subsets from human peripheral blood lymphocytes have been isolated: (1) (E, Fc)- and (2) (E, Ig)-. The first subset was obtained by centrifuging suspensions of macrophage-depleted PBL in which E and EA rosettes had been allowed to form...... population' was shown to be highly variable as judged by the surface markers applied after 4 days of culture, and it is suggested that Null cells contain a number of immature lymphoid cells that may acquire their surface marker during culture. It is concluded that the methods described for purification...

  4. Lymphocytic adenohypophysitis: skull radiographs and MRI

    Energy Technology Data Exchange (ETDEWEB)

    Saiwai, S.; Miyamoto, T. [Department of Radiology, Kobe Central Municipal Hospital, Hyogo (Japan); Inoue, Y.; Nemoto, Y.; Tashiro, T. [Department of Radiology, Osaka City University Medical School (Japan); Ishihara, T. [Department of Endocrinology, Kobe Central Municipal Hospital, Hyogo (Japan); Matsumoto, S. [Department of Neurosurgery, Kobe Central Municipal Hospital, Hyogo (Japan); Hakuba, A. [Department of Neurosurgery, Osaka City University Medical School, 1-5-7 Asahimachi, Abeno, Osaka, 545 (Japan)

    1998-02-01

    We report the skull radiograph, CT and MRI findings in three patients with lymphocytic adenohypophysitis mimicking pituitary adenoma. All cases were associated with pregnancy. CT demonstrated a pituitary mass but did not differentiate lymphocytic adenohypophysitis from pituitary adenoma. The skull radiographs showed either a normal sella turcica or minimal abnormalities; they did not show ballooning or destruction. The MRI appearances were distinctive: relatively low signal on T1-weighted images; preservation of the bright posterior pituitary lobe despite the presence of a relatively large pituitary mass, less common in macroadenomas; marked contrast enhancement compared with pituitary macroadenomas; and dural enhancement adjacent to a pituitary mass. (orig.) With 3 figs., 1 tab., 40 refs.

  5. Effect of steady magnetic field on human lymphocytes

    Energy Technology Data Exchange (ETDEWEB)

    Mileva, M.; Ivanov, B.; Bulanova, M.; Pantev, T.

    1983-01-01

    Exposure to steady magnetic field (SMF) for different periods of time did not elicit statistically reliable increase in chromosome aberrations in human peripheral blood lymphocytes. Metaphase analysis of Crepis capilaris cells revealed that SMF (9 k0e, 200 0e/cm) for 2 days did not induce chromosome aberrations. Nor were any changes demonstrated in roots of beans, onions and L-fibroblasts of subcutaneous tissue of mice and Chinese hamsters. The obtained data are indicative of absence of cytogenetic effect of SMF. The level and spectrum of chromosome aberrations did not exceed the values for spontaneous chromatic fragments in cultures. Cytogenetic analysis of DEDE cells of the Chinese hamster revealed a mild mutagenic effect of SMF. Chromosomal aberrations were also demonstrated after exposure (5 min) of garlic roots.

  6. Fault Analysis and Treatment of MLC VARIAN in Dynamic Intensity Modulated Radiotherapy%动态调强放疗中VARIAN MLC的故障分析处理

    Institute of Scientific and Technical Information of China (English)

    李亮; 陈勇; 徐钰梅; 范雪梅

    2015-01-01

    ObjectiveTo analyze the common faults of 23EX VARIAN linear accelerator MLC and provide the experience for the maintenance of the accelerator. Methods To observe the in the treatment process of the machine failure, observation 4DTC treatment control computer, through the MLC test software to check the fault information to find fault, analysis of the causes of failure, to solve the problem.ResultsAfter scientific analysis, symptomatic treatment, the effective elimination of failure, the system returned to normal operation, maintenance and maintenance for the machine has accumulated experience.Conclusion The use of MLC is high and easy to malfunction. To explore a set of effective method to repair the MLC fault can improve the machine performance, to ensure that the effect of the intensity of radiation therapy.%目的:分析VARIAN 23EX直线加速器MLC常见故障,为加速器维修提供经验交流。方法针对治疗过程中出现机器故障,观察4DTC治疗控制电脑,通过MLC自检软件查看报错信息找到故障所在,分析故障原因,解决故障。结果经过科学分析,对症处理,故障有效排除,系统恢复正常运转,为机器维护维修积累了经验。结论 MLC使用频率高,易发故障。探究一套有效的检修方法及时排除MLC故障可提高机器性能,确保调强放射治疗效果。

  7. Sequential Assessment of Cell Cycle S Phase in Flow Cytometry: A Non-Isotopic Method to Measure Lymphocyte Activation In Vitro

    Directory of Open Access Journals (Sweden)

    Ch. Kohler

    1997-01-01

    Full Text Available Lymphocyte multiplication can be induced in vitro by mitogens or specific antigens, and is usually measured using isotopic methods involving tritiated thymidine. Cellular proliferation can also be analyzed by flow cytometry techniques based on cell cycle analysis through the measurement of DNA content. We applied this method to lymphocytes from 113 individuals, to evaluate lymphocyte proliferation after stimulation in vitro by a mitogen (phytohaemagglutinin, PHA or a recall antigen (tetanus toxoid, using a kinetic approach with four points sequential measurements of the S and G2 phases over six days of culture. The proportion of cells in S phase after PHA stimulation was significantly higher than in controls overall and as early as on day three of the culture. Activation with a recall antigen significantly induced increasing S phase cell proportions up to day six. These data suggest that flow cytometric assessment of the S phase could be a useful alternative to isotopic methods measuring lymphocyte reactivity in vitro.

  8. A double antibody radioimmunoassay for measurement of IgG, IgA and IgM synthesized by human lymphocytes in vitro.

    Directory of Open Access Journals (Sweden)

    Asano,Taro

    1981-11-01

    Full Text Available To investigate cellular interactions between human T and B lymphocytes in various diseases, we established a technique to prove terminal differentiation of B lymphocytes into immunoglobulin synthesizing and secreting cells. We also established a double antibody radioimmunoassay to measure the amount of IgG, IgA and IgM synthesized and secreted in culture supernatants. Purified immunoglobulins were obtained from sera of patients with myeloma or macroglobulinemia. The peripheral blood lymphocytes from 25 normal individuals had the geometric mean synthetic rates of 1886 ng for IgG, 1607 ng for IgA and 1173 ng for IgM per 1 X 10(6 cells when cultured for nine days in the presence of pokeweed mitogen. The method is simple and sensitive, and is thought to be useful for examining human lymphocyte function in vitro.

  9. The Uptake and Utilization of Chlorambucil by Lymphocytes from Patients with Chronic Lymphocytic Leukaemia

    Science.gov (United States)

    Hill, Bridget T.; Harrap, K. R.

    1972-01-01

    It has been shown that lymphocytes isolated from the peripheral blood of patients with chronic lymphocytic leukaemia do not modify the mustard group of chlorambucil, as has been demonstrated previously in Yoshida ascites cells. However, lymphocytes from patients with an unsatisfactory clinical course or poor response to treatment were able to modify the aromatic region of the drug molecule; little change occurred in the aromatic absorption of intracellular chlorambucil in patients who responded to treatment. This simple test may provide a rapid assessment of a patient's potential response to chemotherapy. PMID:4647395

  10. Lymphocytic adrenal medullitis and lymphocytic thyroiditis in a laboratory beagle dog.

    Science.gov (United States)

    Doi, Takuya; Tomonari, Yuki; Kawasako, Kazufumi; Yamada, Naoaki; Tsuchitani, Minoru

    2017-02-04

    Lymphocytic adrenal medullitis characterized by inflammation and atrophy in the medulla of the bilateral adrenal glands was observed in an 18-month-old male laboratory beagle dog. It might be that the present lymphocytic adrenal medullitis is an autoimmune-mediated disease as the histological characteristics are consistent with an autoimmune pathogenesis. However, the actual cause remains unclear as the existence of serum autoantibodies against the adrenal medulla could not be confirmed. Although this dog also contracted lymphocytic thyroiditis along with serum thyroglobulin autoantibodies, indicating that the thyroiditis occurred with an autoimmune basis; the relation between the adrenal medullitis and thyroiditis is unknown.

  11. A novel method for producing target cells and assessing cytotoxic T lymphocyte activity in outbred hosts

    Directory of Open Access Journals (Sweden)

    Bendinelli Mauro

    2009-03-01

    Full Text Available Abstract Background Cytotoxic T lymphocytes play a crucial role in the immunological control of microbial infections and in the design of vaccines and immunotherapies. Measurement of cytotoxic T lymphocyte activity requires that the test antigen is presented by target cells having the same or compatible class I major hystocompatibility complex antigens as the effector cells. Conventional assays use target cells labeled with 51chromium and infer cytotoxic T lymphocyte activity by measuring the isotope released by the target cells lysed following incubation with antigen-specific cytotoxic T lymphocytes. This assay is sensitive but needs manipulation and disposal of hazardous radioactive reagents and provides a bulk estimate of the reporter released, which may be influenced by spontaneous release of the label and other poorly controllable variables. Here we describe a novel method for producing target in outbred hosts and assessing cytotoxic T lymphocyte activity by flow cytometry. Results The method consists of culturing skin fibroblasts, immortalizing them with a replication defective clone of simian virus 40, and finally transducing them with a bicistronic vector encoding the target antigen and the reporter green fluorescent protein. When used in a flow cytometry-based assay, the target cells obtained with this method proved valuable for assessing the viral envelope protein specific cytotoxic T lymphocyte activity in domestic cats acutely or chronically infected with feline immunodeficiency virus, a lentivirus similar to human immunodeficiency virus and used as animal model for AIDS studies. Conclusion Given the versatility of the bicistronic vector used, its ability to deliver multiple and large transgenes in target cells, and its extremely wide cell specificity when pseudotyped with the vesicular stomatitis virus envelope protein, the method is potentially exploitable in many animal species.

  12. In vitro antigenic stimulation of peripheral blood and lymph node lymphocytes of sensitized guinea-pigs: the effect of a second administration of antigen in vivo

    Science.gov (United States)

    Housley, J.; Gell, P. G. H.

    1969-01-01

    Stimulation of DNA synthesis by a guinea-pig albumin—orthanilic acid conjugate (AO) and by tuberculin purified protein derivative (PPD) was obtained in in vitro cultures of peripheral blood and lymph node lymphocytes from guinea-pigs with delayed hypersensitivity to these antigens. Animals sensitized to both AO and PPD were given a further injection of 5 μg AO, intravenously, 8 hours before killing for in vitro studies. In these guinea-pigs, peripheral blood cultures, but not lymph node cultures, showed greater DNA synthesis in response to both AO and PPD than cultures from controls not given a further injection of AO. It is suggested that the further increase in DNA synthesis was due to non-specific lymphocyte `activation' following the interaction of antigen and specifically sensitized lymphocytes. PMID:5352364

  13. Purification of Human Monocytes and Lymphocyte Populations by Counter Current Elutriation– A Short Protocol

    OpenAIRE

    Clarke, Elizabeth V.; Benoit, Marie E.; Tenner, Andrea J.

    2013-01-01

    Investigations of the activation processes involved in human monocytes and monocyte-derived macrophages and dendritic cells often required large numbers of cells that have not been possibly altered or activated by adherence to surfaces, by binding of antibodies to surface antigens during positive selection, or by release of activators by platelets or other non myeloid cells during isolation or co-culture. Human peripheral blood monocytes as well as lymphocytes from the same blood donor can be...

  14. Bone marrow aplasia in B cell chronic lymphocytic leukaemia: successful treatment with antithymocyte globulin.

    OpenAIRE

    Singal, R; Winfield, D A; Greaves, M.

    1991-01-01

    Pure red cell aplasia is a rare but well known association of chronic lymphocytic leukaemia (CLL). Pancytopenia due to bone marrow aplasia has not been previously described in CLL. A 42 year old man with B cell CLL became severely pancytopenic with bone marrow aplasia. Bone marrow culture resulted in a greatly reduced colony formation. High dose corticosteroids and intravenous immunoglobulin treatment were unsuccessful. Prompt and complete marrow recovery ensued after administration of antith...

  15. Loss of signal transduction and inhibition of lymphocyte locomotion in a ground-based model of microgravity

    Science.gov (United States)

    Sundaresan, Alamelu; Risin, Diana; Pellis, Neal R.; McIntire, L. V. (Principal Investigator)

    2002-01-01

    Inflammatory adherence to, and locomotion through the interstitium is an important component of the immune response. Conditions such as microgravity and modeled microgravity (MMG) severely inhibit lymphocyte locomotion in vitro through gelled type I collagen. We used the NASA rotating wall vessel bioreactor or slow-turning lateral vessel as a prototype for MMG in ground-based experiments. Previous experiments from our laboratory revealed that when lymphocytes (human peripheral blood mononuclear cells [PBMCs]) were first activated with phytohemaglutinin followed by exposure to MMG, locomotory capacity was not affected. In the present study, MMG inhibits lymphocyte locomotion in a manner similar to that observed in microgravity. Phorbol myristate acetate (PMA) treatment of PBMCs restored lost locomotory capacity by a maximum of 87%. Augmentation of cellular calcium flux with ionomycin had no restorative effect. Treatment of lymphocytes with mitomycin C prior to exposure to MMG, followed by PMA, restored locomotion to the same extent as when nonmitomycin C-treated lymphocytes were exposed to MMG (80-87%), suggesting that deoxyribonucleic acid replication is not essential for the restoration of locomotion. Thus, direct activation of protein kinase C (PKC) with PMA was effective in restoring locomotion in MMG comparable to the normal levels seen in Ig cultures. Therefore, in MMG, lymphocyte calcium signaling pathways were functional, with defects occurring at either the level of PKC or upstream of PKC.

  16. Loss of signal transduction and inhibition of lymphocyte locomotion in a ground-based model of microgravity

    Science.gov (United States)

    Sundaresan, Alamelu; Risin, Diana; Pellis, Neal R.; McIntire, L. V. (Principal Investigator)

    2002-01-01

    Inflammatory adherence to, and locomotion through the interstitium is an important component of the immune response. Conditions such as microgravity and modeled microgravity (MMG) severely inhibit lymphocyte locomotion in vitro through gelled type I collagen. We used the NASA rotating wall vessel bioreactor or slow-turning lateral vessel as a prototype for MMG in ground-based experiments. Previous experiments from our laboratory revealed that when lymphocytes (human peripheral blood mononuclear cells [PBMCs]) were first activated with phytohemaglutinin followed by exposure to MMG, locomotory capacity was not affected. In the present study, MMG inhibits lymphocyte locomotion in a manner similar to that observed in microgravity. Phorbol myristate acetate (PMA) treatment of PBMCs restored lost locomotory capacity by a maximum of 87%. Augmentation of cellular calcium flux with ionomycin had no restorative effect. Treatment of lymphocytes with mitomycin C prior to exposure to MMG, followed by PMA, restored locomotion to the same extent as when nonmitomycin C-treated lymphocytes were exposed to MMG (80-87%), suggesting that deoxyribonucleic acid replication is not essential for the restoration of locomotion. Thus, direct activation of protein kinase C (PKC) with PMA was effective in restoring locomotion in MMG comparable to the normal levels seen in Ig cultures. Therefore, in MMG, lymphocyte calcium signaling pathways were functional, with defects occurring at either the level of PKC or upstream of PKC.

  17. The effect of ultraviolet radiation on early stages of activation of human lymphocytes: inhibition is independent of effects on DNA

    DEFF Research Database (Denmark)

    Castellanos, G; Owens, T; Rudd, C;

    1982-01-01

    before mitogen was added to the cultures, but were unaffected if irradiation occurred after 16 h of culture in presence of Con A. Cells irradiated with 84 ergs/mm2 at the onset of culture with mitogen did not show the early increase of cation pump function which is a characteristic of stimulated......Low doses (30-84 ergs/mm2, 1 erg = 10(7) J) of ultraviolet radiation (UV) caused severe inhibition of the proliferation of human lymphocytes in vitro. Greatest inhibition was produced when resting cells were irradiated immediately prior to stimulation with concanavalin A (Con A); this was true...... lymphocytes, when this was measured by means of 86Rb uptake after 2-4 h culture. The mitogen-stimulated activation of cation pump function has previously been shown to be unaffected by concentrations of cycloheximide and actinomycin D which produce virtually complete inhibition of protein and RNA synthesis...

  18. Macrophages and lymphocytes differentially modulate the ability of RANTES to inhibit HIV-1 infection.

    Science.gov (United States)

    Gross, Eleanore; Amella, Carol A; Pompucci, Lorena; Franchin, Giovanni; Sherry, Barbara; Schmidtmayerova, Helena

    2003-11-01

    The beta-chemokines MIP-1alpha, MIP-1beta, and RANTES inhibit HIV-1 infection of CD4+ T cells by inhibiting interactions between the virus and CCR5 receptors. However, while beta-chemokine-mediated inhibition of HIV-1 infection of primary lymphocytes is well documented, conflicting results have been obtained using primary macrophages as the virus target. Here, we show that the beta-chemokine RANTES inhibits virus entry into both cellular targets of the virus, lymphocytes and macrophages. However, while virus entry is inhibited at the moment of infection in both cell types, the amount of virus progeny is lowered only in lymphocytes. In macrophages, early-entry restriction is lost during long-term cultivation, and the amount of virus produced by RANTES-treated macrophages is similar to the untreated cultures, suggesting an enhanced virus replication. We further show that at least two distinct cellular responses to RANTES treatment in primary lymphocytes and macrophages contribute to this phenomenon. In lymphocytes, exposure to RANTES significantly increases the pool of inhibitory beta-chemokines through intracellular signals that result in increased production of MIP-1alpha and MIP-1beta, thereby amplifying the antiviral effects of RANTES. In macrophages this amplification step does not occur. In fact, RANTES added to the macrophages is efficiently cleared from the culture, without inducing synthesis of beta-chemokines. Our results demonstrate dichotomous effects of RANTES on HIV-1 entry at the moment of infection, and on production and spread of virus progeny in primary macrophages. Since macrophages serve as a reservoir of HIV-1, this may contribute to the failure of endogenous chemokines to successfully eradicate the virus.

  19. Modulation of Muscle Atrophy, Fatigue and MLC Phosphorylation by MuRF1 as Indicated by Hindlimb Suspension Studies on MuRF1-KO Mice

    Directory of Open Access Journals (Sweden)

    Siegfried Labeit

    2010-01-01

    Full Text Available MuRF1 is a member of the TRIM/RBCC superfamily, a gene family that encompasses a large variety of proteins, all sharing the conserved TRIM (Tripartite Motive sequential array of RING, B-box, and coiled-coil domains. Within this family, MuRF1(also named TRIM63 is a specialized member that contributes to the development of muscle atrophy and sarcopenia. Here we studied MuRF1's role in muscle atrophy during muscle unloading induced by hindlimb suspension. Consistent with previous studies, we found that MuRF1 inactivation leads to an attenuated muscle atrophy response. The amount of protection was higher as compared to the denervation model, and within the 10 day-suspension period the soleus muscle was spared from atrophy in MuRF1-KO mice. Contractility studies on hindlimb suspended muscle tissues suggested that MuRF1's functions extend beyond muscle trophicity and implicate MuRF1 in muscle fatigue and MLC phosphorylation control: soleus muscle from MuRF1-KO mice fatigued significantly faster and in addition showed a reduced posttetanic twitch potentiation. Thus the present work further established the role of MuRF1 in muscle atrophy and for the first time shows that MuRF1 plays a role in muscle fatigue and twitch potentiation.

  20. Impaired NADPH oxidase activity in peripheral blood lymphocytes of galactosemia patients.

    Science.gov (United States)

    Al-Essa, Mazen; Dhaunsi, Gursev S; Al-Qabandi, Wafa'a; Khan, Islam

    2013-07-01

    Galactosemia is an autosomal recessive disorder with a wide range of clinical abnormalities. Cellular oxidative stress is considered as one of the pathogenic mechanisms of galactosemia. In this study, we examined the activity of NADPH oxidase (NOX), a major superoxide-generating enzyme system, in peripheral blood lymphocytes (PBL) from galactosemia patients. PBL were isolated from galactosemia patients and healthy control subjects and used for cell culture studies and biochemical assays. PBL were cultured in the presence or absence of galactose or galactose-1-phosphate (Gal-1-P), and enzyme activities and/or gene expression of NOX, catalase, superoxide dismutase (SOD) and glutathione peroxidase (GPx) were measured in the cell homogenates. PBL isolated from galactosemia patients showed significantly reduced (P Galactosemia patients were found to have significantly (P galactosemia patients; however, Western blotting revealed that NOX-1 protein was not significantly altered. Interestingly, levels of NOX activity in lymphocytes isolated from galactosemia patients significantly increased but remained subnormal when cultured in galactose-deficient medium for two weeks, indicating a galactose-mediated inhibition of NOX. Lymphocytes isolated from control subjects were found to have significantly (P galactosemia patients.

  1. Sulforaphane mitigates genotoxicity induced by radiation and anticancer drugs in human lymphocytes.

    Science.gov (United States)

    Katoch, Omika; Kumar, Arun; Adhikari, Jawahar S; Dwarakanath, Bilikere S; Agrawala, Paban K

    2013-12-12

    Sulforaphane, present in cruciferous vegetables such as broccoli, is a dietary anticancer agent. Sulforaphane, added 2 or 20 h following phytohemaglutinin stimulation to cultured peripheral blood lymphocytes of individuals accidentally exposed to mixed γ and β-radiation, reduced the micronucleus frequency by up to 70%. Studies with whole blood cultures obtained from healthy volunteers confirmed the ability of sulforaphane to ameliorate γ-radiation-induced genotoxicity and to reduce micronucleus induction by other DNA-damaging anticancer agents, such as bleomycin and doxorubicin. This reduction in genotoxicity in lymphocytes treated at the G(0) or G(1) stage suggests a role for sulforaphane in modulating DNA repair. Sulforaphane also countered the radiation-induced increase in lymphocyte HDAC activity, to control levels, when cells were treated 2 h after exposure, and enhanced histone H4 acetylation status. Sulforaphane post-irradiation treatment enhanced the CD 34(+)Lin(-) cell population in culture. Sulforaphane has therapeutic potential for management of the late effects of radiation. Copyright © 2013 Elsevier B.V. All rights reserved.

  2. Dual character of interaction between lymphocytes and allogeneic stem cells

    Energy Technology Data Exchange (ETDEWEB)

    Petrov, R.V.; Dozmorov, I.M.; Kochetkova, M.O.; Nikolaeva, I.S.

    1986-10-01

    The mechanisms of stimulation of colony formation by small doses of allogeneic lymphocytes were studied in mice. When interaction of lymphocytes with allogeneic stem cells was studied, bone marrow cells of mice were injected into lethally irradiated recipients in the control, and mixtures of bone marrow cells with varied numbers of lymphocytes were injected in the experiment. Dependence of the inactivation indices on the number of lymphocytes injected, based on the results of counting macro- and microcolonies in the spleen, is shown.

  3. Morphometric Characterization of Small Cell Lymphocytic Lymphoma

    Directory of Open Access Journals (Sweden)

    Chisoi Anca

    2014-11-01

    Full Text Available The morphometry in histopathology is used to characterize cell populations belonging to different tissues and to identify differences in their parameters with prognostic implications. To achieve morphometric examination were selected 6 of 24 cases identified as small cell lymphocytic lymphoma. For each case analysis was done on five fields, for each field measuring the parameters of 20 cells. The studied parameters were for cytoplasm: cytoplasmic area, maximum and minimum cytoplasmic diameter, cytoplasmic perimeter; for nucleus were measured: nuclear area, minimum and maximum nuclear diameter, nuclear perimeter, nuclear contour index, nuclear ellipticity index, nuclear irregularity index. Also the nucleocytoplasmic ratio was calculated in all studied cases. Small cell lymphocytic lymphoma is characterized in morphometric terms having a small cytoplasmic area (average 29.206 and also a small nuclear area (mean 28.939 having a nucleo-cytoplasmic ratio appearance suggestive for adult lymphocyte. A nuclear contour index small value (3.946, ellipticity index value also small (3.521 and small nuclear irregularity index (3.965. Standard deviations, in any of the studied morphometric categories, is around or below 1 suggesting monomorphic cell appearance. These morphometric and microscopic features characterized mainly by a small population of adult lymphocytes, monomorphic, with rounded hipercromic nuclei, dense chromatin, support the framing into indolent lymphoma group in terms of clinical outcome.

  4. Targeting cytotoxic T lymphocytes for cancer immunotherapy

    OpenAIRE

    Maher, J; Davies, E. T.

    2004-01-01

    In light of their preeminent role in cellular immunity, there is considerable interest in targeting of cytotoxic T-lymphocytes to cancer. This review summarises the active and passive immunotherapeutic approaches under development to achieve this goal, emphasising how recent advances in tumour immunology and gene transfer have impacted upon this field.

  5. Immunophenotypic lymphocyte profiles in human african trypanosomiasis.

    Directory of Open Access Journals (Sweden)

    Caroline Boda

    Full Text Available Human African trypanosomiasis (HAT is a deadly vector-born disease caused by an extracellular parasite, the trypanosome. Little is known about the cellular immune responses elicited by this parasite in humans. We used multiparameter flow cytometry to characterize leukocyte immunophenotypes in the blood and cerebrospinal fluid (CSF of 33 HAT patients and 27 healthy controls identified during a screening campaign in Angola and Gabon. We evaluated the subsets and activation markers of B and T lymphocytes. Patients had a higher percentage of CD19+ B lymphocytes and activated B lymphocytes in the blood than did controls, but lacked activated CD4+ T lymphocytes (CD25+. Patients displayed no increase in the percentage of activated CD8+ T cells (HLA-DR+, CD69+ or CD25+, but memory CD8 T-cell levels (CD8+CD45RA2 were significantly lower in patients than in controls, as were effector CD8 T-cell levels (CD8+CD45RA+CD62L2. No relationship was found between these blood immunophenotypes and disease severity (stage 1 vs 2. However, CD19+ B-cell levels in the CSF increased with disease severity. The patterns of T and B cell activation in HAT patients suggest that immunomodulatory mechanisms may operate during infection. Determinations of CD19+ B-cell levels in the CSF could improve disease staging.

  6. SnapShot: chronic lymphocytic leukemia.

    Science.gov (United States)

    Ciccone, Maria; Ferrajoli, Alessandra; Keating, Michael J; Calin, George A

    2014-11-10

    Chronic lymphocytic leukemia (CLL) is the most common leukemia among adults in western countries. This SnapShot depicts the origins and evolution of this B cell malignancy, describes prognostic factors and CLL animal models, and illustrates therapies in preclinical and clinical development against CLL.

  7. Regulatory T-lymphocytes in asthma

    NARCIS (Netherlands)

    van Oosterhout, AJM; Bloksma, N

    2005-01-01

    T-helper cell type (Th)2 lymphocytes play an important role in the initiation, progression and persistence of allergic diseases, including asthma. However, little is known about immunoregulatory mechanisms that determine susceptibility to, severity of, or persistence of asthma. The concept of a dist

  8. Lymphocyte dynamics in health and disease

    NARCIS (Netherlands)

    van Gent, R.

    2009-01-01

    Following immune depletion, it is vital that the immune system recovers rapidly to avoid severe or life-threatening infections. In adults, full recovery of CD4+ and CD8+ T-cell counts, important cell types of the immune system, may take years. Similar to other lymphocytes, T cells start their develo

  9. Peripheral lymphocyte subpopulations in recurrent aphthous ulceration

    DEFF Research Database (Denmark)

    Pedersen, A; Klausen, B; Hougen, H P

    1991-01-01

    Peripheral lymphocyte subsets--T-helper (CD4+), T-suppressor/cytotoxic (CD8+), and naive/virgin T cells/natural killer cells (CD45RA)--were studied quantitatively in 30 patients with recurrent aphthous ulceration (RAU) and 29 sex- and age-matched RAU-free control donors. The CD4+ percentage was s...

  10. Effect of chloroquine on human lymphocyte proliferation

    DEFF Research Database (Denmark)

    Bygbjerg, Ib Christian; Flachs, H

    1986-01-01

    the response to pokeweed mitogen. The response to concanavalin A and to various antigens was suppressed, especially the response to large particulate antigens. Oral intake of 300 mg of chloroquine base/week did not affect the lymphocyte proliferative responses. 600 mg of base/week decreased the response...

  11. Promutagen activation of triazine herbicides metribuzin and ametryn through Vicia faba metabolism inducing sister chromatid exchanges in human lymphocytes in vitro and in V. faba root tip meristems.

    Science.gov (United States)

    Flores-Maya, Saúl; Gómez-Arroyo, Sandra; Calderón-Segura, María Elena; Villalobos-Pietrini, Rafael; Waliszewski, Stefan M; de la Cruz, Leticia Gómez

    2005-03-01

    The aim of our study was the induction of sister chromatid exchanges (SCE) in human lymphocytes in vitro and in root tip meristems of Vicia faba to evaluate the genotoxic effects of metribuzin and ametryn. Direct treatments of these herbicides on human lymphocytes in vitro applied 24 h after the beginning of culture did not induce SCE; however, they showed a cytotoxic effect in the cultures expressed as cellular death. On the contrary, when extracts of V. faba roots, treated for 4 h with metribuzin and ametryn (in vivo activation), were added to the lymphocyte cultures, SCEs were significantly induced with an asymptotic response. Negative responses appeared with the in vitro assays, in which metribuzin and ametryn were added directly to the 48 h lymphocyte cultures for 4 h. Nevertheless, in treatments in which the S10 metabolic mix was added, the SCE frequencies were significantly different to the control, although a concentration-response relationship was only observed with metribuzin. The results showed that both herbicides needed the V. faba metabolism to produce SCE in human lymphocyte cultures. Metribuzin and ametryn applied to V. faba root tip meristems for 4 h increased SCE frequency significantly, and a concentration-response relationship was observed with both herbicides.

  12. Purple patches in an immunocompromised patient: a report of secondary disseminated cutaneous mucormycosis in a man with chronic lymphocytic leukemia.

    Science.gov (United States)

    Iyengar, Sanjana; Chambers, Cindy J; Millsop, Jillian W; Fung, Maxwell A; Sharon, Victoria R

    2017-03-15

    A 60-year-old man with chronic lymphocytic leukemiadeveloped a deeply violaceous annular patchwith a halo of erythema on the right thigh duringhospitalization for neutropenic fever. Associatedsymptoms included chronic cough and fatigue.Bilateral lung opacities with hilar lymphadenopathywere noted on chest computed tomographyscan. Punch biopsy and tissue culture confirmeda diagnosis of secondary disseminated cutaneousmucormycosis. Although rare, physicians shouldinclude mucormycosis in the differential diagnosisof purpuric patches in immunosuppressed patients.Prompt skin biopsy and tissue culture may optimizethe success of treatment.

  13. Rosette formation of pig T lymphocytes with sheep erythrocytes.

    Science.gov (United States)

    Escajadillo, C; Binns, R M

    1975-01-01

    The relationship of sheep RBC rosette formation to density of thymus and blood lymphocytes was investigated. Thymocyte density was unimodal and cells of all densities rosetted equally. Blood lymphocyte density was bimodal with most rosette-forming cells in the denser ficoll layers. Papain treatment of SRBC increases rosette formation with blood lymphocytes while apparently maintaining specificity of T cells.

  14. 21 CFR 864.8500 - Lymphocyte separation medium.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Lymphocyte separation medium. 864.8500 Section 864.8500 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED... medium. (a) Identification. A lymphocyte separation medium is a device used to isolate lymphocytes from...

  15. DMPD: Developmental plasticity of lymphocytes. [Dynamic Macrophage Pathway CSML Database

    Lifescience Database Archive (English)

    Full Text Available 18472258 Developmental plasticity of lymphocytes. Cobaleda C, Busslinger M. Curr Op...in Immunol. 2008 Apr;20(2):139-48. Epub 2008 May 9. (.png) (.svg) (.html) (.csml) Show Developmental plastic...ity of lymphocytes. PubmedID 18472258 Title Developmental plasticity of lymphocytes. Authors Cobaleda C, Bus

  16. Chronic lymphocytic leukemia/small lymphocytic lymphoma presenting as septic arthritis of the shoulder

    Energy Technology Data Exchange (ETDEWEB)

    Donovan, Andrea; Schweitzer, Mark E.; Nomikos, George [NYU Hospital for Joint Diseases, New York, NY (United States); Garcia, Roberto A. [Bellevue Hospital Center, New York, NY (United States)

    2008-11-15

    We report a case of a 53-year-old man presenting with shoulder pain mimicking septic arthritis. Laboratory findings were atypical. Biopsy performed to assess for possible osteomyelitis demonstrated chronic lymphocytic leukemia/small lymphocytic lymphoma. Intra-articular lymphoma is a rare but important consideration in patients with atypical clinical presentation. Imaging alone may be insufficient to render diagnosis as lymphoma can mimic infection, synovial hypertrophic processes, and depositional arthropathy. (orig.)

  17. NEUTROPHIL/LYMPHOCYTE RATIO AND PLATELET/LYMPHOCYTE RATIO IN PATIENTS WITH NSCLC

    OpenAIRE

    Cukic, Vesna

    2016-01-01

    Objective: to compare neutrophil/lymphocyte ratio (NLR) and platelet/lymphocyte ratio (PLR) in patients with NSCLC (Non- Small- Cell Lung Cancer): with and without metastases at the time of diagnosis to find out if there is the importance of these cell ratios in the assessment of severity NSCLC. Material and Methods: this is the retrospective analysis of NRL and PRL in patients with NSCLC at the time of the diagnosis of disease before any anti tumor treatment (chemotherapy, radiotherapy, surg...

  18. Lymphocyte transformation studies in drug hypersensitivity.

    Science.gov (United States)

    Warrington, R J; Tse, K S

    1979-05-05

    In a group of patients with clinically diagnosed drug hypersensitivity the in vitro lymphocyte response to the suspected drug was assessed by the lymphocyte transformation test. The test gave positive results in all 15 patients with penicillin-induced immediate or accelerated allergic reactions and positive immediate skin-test reactivity to the major or the minor antigenic determinant of penicillin, or both, but in only 3 of the 12 patients with delayed-onset maculopapular rashes induced by penicillin, despite positive immediate reactivity to the skin-test reagents.Lymphocyte stimulation greater than five times the control level was demonstrated for five patients with penicillin-induced erythroderma, Stevens-Johnson syndrome or a serum-sickness-like illness, or with methicillin-induced interstitial nephritis, all of whom had negative reactions to the appropriate skin-test reagents. A low level of stimulation was seen in eight other skin-test-negative patients with possible allergic reactions induced by penicillins. However, in all subjects tested the stimulation was significantly greater than the mean for control subjects.For 9 of 11 patients with isoniazid-induced hepatitis or maculopapular rashes, but for only 8 of 31 patients with eruptions induced by a variety of drugs other than penicillins and isoniazid, significant stimulation occurred in the lymphocyte transformation test.It is concluded that the lymphocyte transformation test is useful in the detection of hypersensitivity to the penicillins (although in IgE-mediated reactions skin testing is clearly preferable) and isoniazid but is of limited value in the demonstration of hypersensitivity to other drugs.

  19. Clonal priming of human lymphocytes: Specificity and cross-reactivity of cellular immune reactions.

    Science.gov (United States)

    Levis, W R; Datiner, A M

    1977-04-23

    Clonal priming in response to chemical and microbial antigens which defines the specificity of cellular immune reactions, was demonstrated by culture techniques. Human leucocyte cultures stimulated with specific antigens typically show peak levels of D.N.A. synthesis after 5 to 7 days in culture. Such primary leucocyte cultures were incubated for 10-20 days, then the cells were gently centrifuged and resuspended in fresh RPMI 1640 with 20% plasma. These secondary or primed cultures typically showed less than 1000 c.p.m. after 48 hours. However, if the original antigenic stimulant was added, specific accelerated responses were seen by 48 hours in the secondary cultures. Lymphocyte clones in these sceondary cultures primed with dinitrophenylated (D.N.P.) antigens (from subjects sensitised to dinitrochlorobenzene) showed enhanced D.N.A. sythesis in response to the same dinitrophenylated antigens and showed varible accelerated responses to related chemically modified antigens. However, D.N.P.-activated clones in these secondary cultures did not show enhanced responses to microbial antigens even though the lymphocytes had been highly responsive to tetanus toxoid and other microbial antigens in primary cultures. The specificity of this clonal activation was further demonstrated by the enhanced response of secondary cultures of tetanus-toxoid-activated clones to tetanus toxoid but not to dinitrophenylated antigens. The abiltty to detect specificity and cross-reactivity of cellular immune reaction has broad implications for investigations of cellular immunity as well as many potential applications in the diagnosis and understanding the patogenesis of inflammatory and neoplastic diseases in which cellular immune discrimination may be involved.

  20. Protection Against Lung Cancer Patient Plasma-Induced Lymphocyte Suppression by Ganoderma Lucidum Polysaccharides

    Directory of Open Access Journals (Sweden)

    Li-Xin Sun

    2014-01-01

    Full Text Available Background/Aims: This study was conducted to determine the potential of Ganoderma lucidum polysaccharides (Gl-PS in protection against lung cancer patient plasma-induced suppression of lymphocytes. Lung cancer is a major cause of disease and loss of life in the United States and worldwide. Cancer cells release immunosuppressive mediators, such as PGE2, TGF-β, IL-10, and VEGF, to inhibit the immune response to escape from immune surveillance. Gl-PS has been shown to counteract this immune inhibition in an animal cell culture model, and thus to facilitate tumor control. The present study explored whether or not such an effect could also be demonstrated in human lung cancer patients. Methods: Immunofluorescence, flow cytometry, MTT, immunocytochemistry, and western blot analysis were used to assess lymphocyte activation with PHA. Results: The plasma of lung cancer patients suppressed proliferation, CD69 expression, and perforin and granzyme B production in lymphocytes upon activation by PHA, effects that were partially of fully reversed by Gl-PS. Conclusion: Lung cancer patient plasma-induced suppression of lymphocyte activation by phytohemagglutinin may be antagonized fully or partially by Gl-PS, an observation suggesting the potential of Gl-PS in cancer therapy.

  1. Is lymphocytic (hashimoto) thyroiditis associated with suicide?

    Science.gov (United States)

    Cina, Stephen J; Perper, Joshua A

    2009-09-01

    The histologic diagnosis of lymphocytic (Hashimoto) thyroiditis requires lymphocytic inflammation of the thyroid gland in combination with Hourthle cell metaplasia of follicular epithelial cells. Clinically, this autoimmune process has been associated with hypothyroidism and psychiatric conditions including depression. This retrospective study was designed to quantify the incidence and severity of lymphocytic thyroiditis in a series of nonconsecutive suicides compared with a cohort of motor vehicle accident victim controls. Eighty-one suicide victims (61 male, 20 female; age range 13-79 years, average 43) were compared with 88 age and gender matched controls (64 males, 24 females; age range 19-85 years, average 36). The degree of lymphocytic inflammation of the thyroid gland was graded on a scale of 0 to 3 (0 = no inflammation, 1 = mild inflammation, 2-3 moderate-to-marked inflammation with Hourthle cell metaplasia). Slides from each case were reviewed while blinded to the cause and manner of death in each case. Of these 169 total cases, 8 (4.7%) received a score of 3, whereas additional 7 (4.1%) received a grade of 2. Eighty-six percent of all of the cases showed no significant inflammation and recorded a score of 0. Of the 81 suicides, 3 had a score of 3, and 3 had a score of 2 (combined incidence of 7.4%). Within the control group, 5 of 88 cases scored 3 and another 4 scored 2 (combined incidence = 10.2%). Three males and 5 females scored 3 with an age range of 23 to 63 years, average 42. Incidental data tabulated showed that 19% of suicide victims were on psychoactive medications compared with 6% in the motor vehicle accident control group. No one on this study was on thyroid hormone replacement therapy. Depression is strongly linked to suicide and lymphocytic thyroiditis may be a cause of depression. Based on this study, however, the presence of lymphocytic thyroiditis cannot be used as a histologic adjunct to discriminate between suicide and accident in

  2. Carbon nanotube-mediated delivery of nucleic acids does not result in non-specific activation of B lymphocytes

    Energy Technology Data Exchange (ETDEWEB)

    Cai Dong [Department of Biology, Boston College, Chestnut Hill, MA 02467 (United States); Doughty, Cheryl A [Department of Biology, Boston College, Chestnut Hill, MA 02467 (United States); Potocky, Terra B [Department of Biology, Boston College, Chestnut Hill, MA 02467 (United States); Dufort, Fay J [Department of Biology, Boston College, Chestnut Hill, MA 02467 (United States); Huang Zhongping [NanoLab, Incorporated, Newton, MA 02458 (United States); Blair, Derek [Department of Biology, Boston College, Chestnut Hill, MA 02467 (United States); Kempa, Krzysztof [Department of Physics, Boston College, Chestnut Hill, MA 02467 (United States); Ren, Z F [Department of Physics, Boston College, Chestnut Hill, MA 02467 (United States); Chiles, Thomas C [Department of Biology, Boston College, Chestnut Hill, MA 02467 (United States)

    2007-09-12

    The efficient delivery of genes and proteins into primary mammalian cells and tissues has represented a formidable challenge. Recent advances in the research of carbon nanotubes (CNTs) offer much promise for their use as delivery platforms into mammalian cells. Ideally, CNT-mediated applications should not result in cellular toxicity nor perturb cellular homeostasis (e.g., result in non-specific activation of primary cells). It is therefore critical to evaluate the impact of CNT exposure on the cellular metabolism, proliferation and survival of primary mammalian cells. We investigated the compatibility of a recently developed CNT-mediated delivery method, termed nanospearing, with primary ex vivo cultures of B lymphocytes. Several parameters were evaluated to assess the impact of CNTs on naive B lymphocytes, including cell survival, activation, proliferation and intracellular signal transduction. Our results indicate that nanospearing does not result in the activation of naive primary B lymphocytes nor alter survival in ex vivo cultures. Herein, B cells exposed to CNTs were capable of responding to extrinsic pro-survival signals such as interleukin-4 and signaling by the B-cell antigen receptor in a manner similar to that of B cells cultured in the absence of CNTs. Our study demonstrates the biocompatibility of the CNT-mediated nanospearing procedure with respect to primary B lymphocytes.

  3. Lymphocyte trafficking and HIV infection of human lymphoid tissue in a rotating wall vessel bioreactor

    Science.gov (United States)

    Margolis, L. B.; Fitzgerald, W.; Glushakova, S.; Hatfill, S.; Amichay, N.; Baibakov, B.; Zimmerberg, J.

    1997-01-01

    The pathogenesis of HIV infection involves a complex interplay between both the infected and noninfected cells of human lymphoid tissue, the release of free viral particles, the de novo infection of cells, and the recirculatory trafficking of peripheral blood lymphocytes. To develop an in vitro model for studying these various aspects of HIV pathogenesis we have utilized blocks of surgically excised human tonsils and a rotating wall vessel (RWV) cell culture system. Here we show that (1) fragments of the surgically excised human lymphoid tissue remain viable and retain their gross cytoarchitecture for at least 3 weeks when cultured in the RWV system; (2) such lymphoid tissue gradually shows a loss of both T and B cells to the surrounding growth medium; however, this cellular migration is reversible as demonstrated by repopulation of the tissue by labeled cells from the growth medium; (3) this cellular migration may be partially or completely inhibited by embedding the blocks of lymphoid tissue in either a collagen or agarose gel matrix; these embedded tissue blocks retain most of the basic elements of a normal lymphoid cytoarchitecture; and (4) both embedded and nonembedded RWV-cultured blocks of human lymphoid tissue are capable of productive infection by HIV-1 of at least three various strains of different tropism and phenotype, as shown by an increase in both p24 antigen levels and free virus in the culture medium, and by the demonstration of HIV-1 RNA-positive cells inside the tissue identified by in situ hybridization. It is therefore reasonable to suggest that gel-embedded and nonembedded blocks of human lymphoid tissue, cocultured with a suspension of tonsillar lymphocytes in an RWV culture system, constitute a useful model for simulating normal lymphocyte recirculatory traffic and provide a new tool for testing the various aspects of HIV pathogenesis.

  4. Protective effect of apigenin on radiation-induced chromosomal damage in human lymphocytes

    Science.gov (United States)

    Rithidech, Kanokporn Noy; Tungjai, Montree; Whorton, Elbert B.

    2005-01-01

    The potential use of flavonoids as a radioprotector is of increasing interest because of their high antioxidant activity and abundance in the diet. The aim of this study is to examine genotoxic and radioprotective effects of one of the most common flavonoids, apigenin, on radiation-induced chromosome aberrations in human lymphocytes. The cytokinesis-block micronucleus (CBMN) assay was used to evaluate such effects of apigenin. Blood samples were collected from two non-smoking healthy male volunteers who had no history of previous exposure to other clastogenic agents. Isolated lymphocytes were cultured. There were two tubes per concentration for all treatments. To evaluate the genotoxicity of apigenin, cells were first treated with different concentrations of apigenin (0, 2.5, 5, 10 and 25 microg/mL) at 24 h after culture initiation, followed by cytochalasin-B (Cyt-B) treatment (3 microg/mL) and cell harvest at 44 and 72 h, respectively. Secondly, to investigate the radioprotective effect, cell cultures were exposed to different concentrations of apigenin as described above for 30 min before being irradiated to 2 Gy of 137Cs gamma rays (at a dose rate of 0.75 Gy/min). In all instances, the frequency of MN was scored in binucleated (BN) cells. The nuclear proliferation index also was calculated. We did not detect an increase in the frequency of MN in non-irradiated human lymphocyte cultures treated with 2.5, 5.0 or 10 microg/mL apigenin; although, we did observe an increase in cultures treated with 25 microg/mL apigenin (the highest concentration of apigenin used in our study). We also observed a significant increase in the frequency of MN in irradiated cells overall; however, the frequency was decreased as the concentration of apigenin increased, suggesting a radioprotective effect. These findings provide a basis for additional studies to help clarify the potential use and benefit of apigenin as a radioprotector.

  5. Evaluation of CD4+CD25+ T lymphocyte response time kinetics in patients with chronic Chagas disease after in vitro stimulation with recombinant Trypanosoma cruzi antigens

    Directory of Open Access Journals (Sweden)

    Suellen Carvalho de Moura Braz

    2013-05-01

    Full Text Available Introduction CD4+CD25+ T lymphocytes have been implicated in the regulation of host inflammatory response against Trypanosoma cruzi, and may be involved in the clinical course of the disease. Methods Peripheral blood mononuclear cells from patients with chronic Chagas disease were cultured in the presence of T. cruzi recombinant antigens and assayed for lymphocytes at distinct time points. Results It was possible to differentiate clinical forms of chronic Chagas disease at days 3 and 5 according to presence of CD4+CD25+ T cells in cell cultures. Conclusions Longer periods of cell culture proved to be potentially valuable for prospective evaluations of CD4+CD25+ T lymphocytes in patients with chronic Chagas disease.

  6. Mean dose to lymphocytes during radiotherapy treatments

    Energy Technology Data Exchange (ETDEWEB)

    Brandan, M.E.; Perez-Pastenes, M.A. [Instituto de Fisica (Mexico); Ostrosky-Wegman, P.; Gonsebatt, M.E. [Instituto de Investigaciones Biomedicas (Mexico); Diaz-Perches, R. [Hospital General de Mexico (Mexico)

    1994-10-01

    Using a probabilistic model with parameters from four radiotherapy protocols used in Mexican hospitals for the treatment of cervical cancer, the authors have calculated the distribution of dose to cells in peripheral blood of patients. Values of the mean dose to the lymphocytes during and after a {sup 60}Co treatment are compared to estimates from an in vivo chromosome aberration study performed on five patients. Calculations indicate that the mean dose to the circulating blood is about 2% of the tumor dose, while the mean dose to recirculating lymphocytes may reach up to 7% of the tumor dose. Differences up to a factor of two in the dose to the blood are predicted for different protocols delivering equal tumor doses. The data suggest mean doses higher than the predictions of the model. 10 refs., 3 figs., 2 tabs.

  7. Cell Death Mechanisms Induced by Cytotoxic Lymphocytes

    Institute of Scientific and Technical Information of China (English)

    Ch(a)vez-Gal(a)n L; Arenas-Del Angel MC; Zenteno E; Ch(a)vez R; Lascurain R

    2009-01-01

    One of the functions of the immune system is to recognize and destroy abnormal or infected cells to maintain homeostasis. This is accomplished by cytotoxic lymphocytes. Cytotoxicity is a highly organized multifactor process. Here, we reviewed the apoptosis pathways induced by the two main cytotoxic lymphocyte subsets, natural killer (NK) cells and CD8+T cells. In base to recent experimental evidence, we reviewed NK receptors involved in recognition of target-cell, as well as lytic molecules such as perforin, granzymes-A and -B, and granulysin. In addition, we reviewed the Fas-FasL intercellular linkage mediated pathway, and briefly the cross-linking of tumor necrosis factor (TNF) and TNF receptor pathway. We discussed three models of possible molecular interaction between lyric molecules from effector cytotoxic cells and target-cell membrane to induction of apoptosis.

  8. Bioluminescent assay for human lymphocyte blast transformation.

    Science.gov (United States)

    Bulanova, E G; Budagyan, V M; Romanova, N A; Brovko LYu; Ugarova, N N

    1995-05-01

    One of the basic tests of in vitro evaluation of immune cell functional activity is a proliferative response of lymphocytes on the action of external stimuli such as mitogenic lectines, antigens, etc. We compared two methods used to assess the lymphocyte functional status. (1) [3H]thymidine incorporation and (2) bioluminescence for determination of intracellular ATP in blast cells. Comparison has been done for healthy donors and patients with proven low immunological status. The proposed bioluminescent method for evaluation of the proliferative response was shown to be sensitive enough for diagnostic purposes. This method allows one to process a large number of samples at the same time and correlates highly with the radionuclide test use hazardous radioactive materials.

  9. Lymphocyte transformation in presumed ocular histoplasmosis

    Energy Technology Data Exchange (ETDEWEB)

    Ganley, J.P.; Nemo, G.J.; Comstock, G.W.; Brody, J.A.

    1981-08-01

    Lymphocytes from individuals with inactive macular disciform lesions of presumed ocular histoplasmosis challenged with three histoplasmin antigens incorporated tritiated thymidine at a significantly higher rate than histoplasmin-stimulated lymphocytes of matched control and peripheral scar groups. This finding is consistent with the etiologic association of the disciform ocular syndrome and previous systemic infection with Histoplasma capsulatum. The disciform group had a higher mean response than the other two groups to pokeweed mitogen but not to phytohemagglutinin and had higher mean counts per minute to the specific antigens Toxoplasma gondii, Blastomyces dermatitidis, Cryptococcus neoformans, Mycobacterium tuberculosis, M battery, and M gaus, but not to Candida albicans. These data would suggest that individuals with the disciform lesion of presumed ocular histoplasmosis have a hyperreactive cellular immune response; this response may play an important role in the development of the disciform.

  10. Obinutuzumab for previously untreated chronic lymphocytic leukemia.

    Science.gov (United States)

    Abraham, Jame; Stegner, Mark

    2014-04-01

    Obinutuzumab was approved by the Food and Drug Administration in late 2013 for use in combination with chlorambucil for the treatment of patients with previously untreated chronic lymphocytic leukemia (CLL). The approval was based on results of an open-label phase 3 trial that showed improved progression-free survival (PFS) with the combination of obinutuzumab plus chlorambucil compared with chlorambucil alone. Obinutuzumab is a monoclonal antibody that targets CD20 antigen expressed on the surface of pre B- and mature B-lymphocytes. After binding to CD20, obinutuzumab mediates B-cell lysis by engaging immune effector cells, directly activating intracellular death signaling pathways, and activating the complement cascade. Immune effector cell activities include antibody-dependent cellular cytotoxicity and antibody-dependent cellular phagocytosis.

  11. The Danish National Chronic Lymphocytic Leukemia Registry

    DEFF Research Database (Denmark)

    da Cunha-Bang, Caspar; Geisler, Christian Hartmann; Enggaard, Lisbeth;

    2016-01-01

    , and for decision on treatment initiation as well as characteristics included in the CLL International Prognostic Index are collected. DESCRIPTIVE DATA: To ensure full coverage of Danish CLL patients in the registry, both continuous queries in case of missing data, and cross-referencing with the Danish National......, 3,082 patients have been registered. CONCLUSION: The Danish National CLL Registry is based within the Danish National Hematology Database. The registry covers a cohort of all patients diagnosed with CLL in Denmark since 2008. It forms the basis for quality assessment of CLL treatment in Denmark......AIM: In 2008, the Danish National Chronic Lymphocytic Leukemia Registry was founded within the Danish National Hematology Database. The primary aim of the registry is to assure quality of diagnosis and care of patients with chronic lymphocytic leukemia (CLL) in Denmark. Secondarily, to evaluate...

  12. Metabolism pathways in chronic lymphocytic leukemia.

    Science.gov (United States)

    Rozovski, Uri; Hazan-Halevy, Inbal; Barzilai, Merav; Keating, Michael J; Estrov, Zeev

    2016-01-01

    Alterations in chronic lymphocytic leukemia (CLL) cell metabolism have been studied by several investigators. Unlike normal B lymphocytes or other leukemia cells, CLL cells, like adipocytes, store lipids and utilize free fatty acids (FFA) to produce chemical energy. None of the recently identified mutations in CLL directly affects metabolic pathways, suggesting that genetic alterations do not directly contribute to CLL cells' metabolic reprogramming. Conversely, recent data suggest that activation of STAT3 or downregulation of microRNA-125 levels plays a crucial role in the utilization of FFA to meet the CLL cells' metabolic needs. STAT3, known to be constitutively activated in CLL, increases the levels of lipoprotein lipase (LPL) that mediates lipoprotein uptake and shifts the CLL cells' metabolism towards utilization of FFA. Herein, we review the evidence for altered lipid metabolism, increased mitochondrial activity and formation of reactive oxygen species (ROS) in CLL cells, and discuss the possible therapeutic strategies to inhibit lipid metabolism pathways in patient with CLL.

  13. B cell acute lymphocytic leukemia in pregnancy.

    Science.gov (United States)

    Bottsford-Miller, Justin; Haeri, Sina; Baker, Arthur M; Boles, Jeremiah; Brown, Mark

    2011-08-01

    Acute lymphocytic leukemia (ALL) is a rare occurrence in pregnancy and can be rapidly fatal if left untreated. The need for immediate treatment of ALL, coupled with the maternal-fetal risks from the chemotherapy regimen render a therapeutic dilemma in pregnant women with ALL. We report a case of ALL diagnosed in the 24th week of pregnancy to outline our management strategy, to demonstrate the feasibility of treatment with multi-agent chemotherapy, and to provide a review of the literature.

  14. GABA, a natural immunomodulator of T lymphocytes

    DEFF Research Database (Denmark)

    Bjurstöm, Helen; Wang, Junyang; Ericsson, Ida

    2008-01-01

    gamma-aminobutyric acid (GABA) is the main neuroinhibitory transmitter in the brain. Here we show that GABA in the extracellular space may affect the fate of pathogenic T lymphocytes entering the brain. We examined in encephalitogenic T cells if they expressed functional GABA channels that could......M and higher GABA concentrations decreased T cell proliferation. The results are consistent with GABA being immunomodulatory....

  15. A galactose-inhibitable mitogen for human lymphocytes from the sponge axinella polypoides.

    Science.gov (United States)

    Phillips, S G; Bretting, H; Kabat, E A

    1976-10-01

    Of two galactose-binding hemagglutinins isolated from the sponge Axinella polypoides, axinella I was strongly mitogenic for human peripheral blood lymphocytes, and axinella II was not. Purified T cells responded strongly and B cells weakly to axinella I. Mitogenic response, as monitored by rate of 3H-thymidine incorporation on the third day of culture, was specifically inhibited by Dgalactose, Dfucose, raffinose, or 2-deoxy-D-galactose added within 5 hr of the mitogen. Mitogenic response was correlated with degree of lymphocyte agglutination. The effectiveness of a given sugar in inhibiting mitogenic response to axinella I paralleled its potency in inhibiting precipitation of lectin by blood group substances. If an inhibitory concentration of Dgalactose was add 24 to 40 hr after mitogenic activation, rate of 3H-thymadine uptake at 72 hr was two to twenty times above the rate induced in cultures to which no galactose was added. Dgalactose at a subinhibitory concentration (10mug/ml) enhanced 3H-thymidine incorportion incorporation induced by phytohemagglutinin or Con A, an effect reversible by Dgalactose. These findings suggest that axinella I has tow antagonistic effects on human lymphocytes: a) mitogenic activation and b) depressive activity resulting from depletion of essential galactose moieties.

  16. Lymphocytic hypophysitis masquerading as pituitary adenoma

    Directory of Open Access Journals (Sweden)

    Rajneesh Mittal

    2012-01-01

    Full Text Available Introduction: Pituitary hypophysitis (PH is characterized by pituitary infiltration of lymphocytes, macrophages, and plasma cells that could lead to loss of pituitary function. Hypophysitis may be autoimmune or secondary to systemic diseases or infections. Based on the histopathological findings PH is classified into lymphocytic, granulomatous, xanthomatous, mixed forms (lymphogranulomatous, xanthogranulomatous, necrotizing and Immunoglobulin- G4 (IgG4 plasmacytic types. Objective: To report a case of lymphocytic hypophysitis (LH. Case Report: A 15-year-old girl presented with history of headache, amenorrhea, and history of polyuria for past 4 months. Initial evaluation had suppressed follicular stimulating hormone (<0.01 mIU/ml, high prolactin levels (110.85 ng/ml and diabetes insipidus (DI. Magnetic resonance imaging of sella was suggestive of pituitary macroadenoma with partial compression over optic chiasma. Patient underwent surgical decompression. Yellowish firm tissue was evacuated and xanthochromic fluid was aspirated. Histopathology was suggestive of LH. She resumed her cycles postoperatively after 4 months, prolactin levels normalized, however, she continues to have DI and is on desmopressin spray. This case has been presented here for its rare presentation in an adolescent girl because it is mostly seen in young females and postpartum period and its unique presentation as an expanding pituitary mass with optic chiasma compression. Conclusion: Definitive diagnosis of LH is based on histopathological evaluation. Therapeutic approach should be based on the grade of suspicion and clinical manifestations of LH.

  17. [Circadian rhythm of human lymphocyte subpopulations].

    Science.gov (United States)

    Pasqualetti, P; Colantonio, D; Casale, R; Colangeli, S; Natali, G

    1988-01-01

    Circadian rhythm of lymphocyte subsets was investigated in four healthy subjects, males, aged 35-58 years old. After a period of ambiental synchronization, venous blood samples were taken during a span of a day at 0.00 a.m., 4.00 a.m., 8.00 a.m., noon, 4.00 p.m. and 8.00 p.m. Lymphocyte subsets (OKT3, OKT4, OKT8, OKB7, OKJa1) were determined by monoclonal antibodies method, and serum level of cortisol by radioimmunoassay method. The OKT4/OKT8 ratio was also calculated. Data were analyzed by chronograms (mean +/- 1SD) and by cosinor method. Results show a significant circadian rhythm for each lymphocyte subset and for serum cortisol levels. The lowest levels of all circulating subsets were seen between noon and 4.00 p.m. and the highest levels around midnight, inversely related with the circadian rhythm of serum cortisol. The OKT4/OKT8 ratio, on the contrary, was relatively constant during the day, without a significant circadian rhythm. These observations have laboratoristic, clinical, and therapeutic implications and should be considered in the course of immunological studies.

  18. Normal lymphocyte immunophenotype in an elderly population

    Directory of Open Access Journals (Sweden)

    Sâmia Macedo Queiroz Mota Castellão Tavares

    2014-06-01

    Full Text Available OBJECTIVE: The aim of this work was to evaluate the lymphocyte immunophenotype in an elderly population.METHODS: This study enrolled 35 over 60-year-old volunteers and a control group composed of 35 young adults. The study included elderly without diseases that might affect the functioning of the immune system. These individuals were consulted by doctors and after a physical examination, laboratory tests were performed using a Beckman Coulter (r flow cytometer. The GraphPad Prism computer program was employed for statistical analysis with the level of significance being set for p-values <0.05.RESULTS: There is a statistically significant reduction in the number of lymphocytes (CD8 +, CD2 + and CD3 + cells in the elderly compared to young adults. These low rates are explained by changes attributed to aging and may be partly responsible for the reduction in the cellular immune response, lower proliferative activity and the low cytotoxicity of lymphocytes.CONCLUSION: These parameters showed greater impairment of adaptive immunity in the elderly population and can therefore explain the greater fragility of the aged body to developing diseases.

  19. Sudden unexpected death associated with lymphocytic thyroiditis.

    Science.gov (United States)

    Vestergaard, Vibeke; Drostrup, Dorthe Høj; Thomsen, Jørgen L

    2007-04-01

    A forensic autopsy study comprising 125 cases was carried out retrospectively in order to evaluate pathological changes in the thyroid gland in different groups of death. The five groups selected consecutively were: (i) opiate addicts who died from an overdose, (ii) alcoholics who died as a result of their alcohol abuse, (iii) cases of fatal poisoning other than opiate addicts, (iv) unknown cause of death and (v) controls without prior disease. Tissue samples from the thyroid gland were cut and stained with haematoxylin and eosin and van Gieson. Histology examinations were subsequently performed blind with semiquantitative assessment of the following six parameters: (a) height of the follicular epithelium, (b) the amount of lymphocytes, (c) the presence of plasma cells, (d) hyperplastic follicular changes, (e) oxyphilic changes, and (f) fibrosis. The most striking result was the finding of extensive lymphocytic infiltration of the thyroid parenchyma in five of the 124 cases, of which four belonged in the group of 'unknown cause of death'. This discovery leads to reflections regarding lymphocytic thyroiditis as a cause of death, either by itself or in combination with other disorders. Silent (painless) thyroiditis, especially, is easily overlooked at autopsy as there are no macroscopic changes and often no prior symptoms or history of thyroid disease pointing towards this condition. Analyses of thyroid hormones are unreliable in predicting endocrine status in life. Routine microscopy of the thyroid gland is therefore advocated in cases of sudden unexpected death in order to diagnose thyroid disease, in particular silent (painless) thyroiditis.

  20. Metaphase yields from staphylococcal enterotoxin A stimulated peripheral blood lymphocytes of unirradiated and irradiated aged rhesus monkeys

    Energy Technology Data Exchange (ETDEWEB)

    Hill, F.S.; Cantu, A.O.; Lucas, J.N. (Lawrence Livermore National Lab., CA (United States)); Cox, A.B.; Salmon, Y.L. (Air Force Armstrong Lab., Brookes AFB, TX (United States))

    1994-10-01

    The mitogen phytohemagglutinin (PHA) works well in both human and cynomolgus monkey (Macaca fascicularis) lymphocyte cultures to stimulate T cell proliferation. T cells from rhesus monkeys (Macaca mulatta) are less responsive than human cells, producing few metaphases when thousands are required, e.g. in biological dosimetry studies. We show that staphylococcal enterotoxin A (SEA), one of the most potent mitogens known, at a concentration of 0.5 [mu]g/ml stimulated peripheral lymphocytes to grow with a mitotic index (MI) averaging 0.13 metaphases/cell in old, irradiated rhesus macaques. This was significantly greater (p < 0.001) that that produced by PHA (M1<0.01) in lymphocytes from the same animals. Whole blood was cultured for 96, 120 and 144 h for five irradiated individuals and for two controls. All cells cultured with SEA produced a high MI with a peak response at 120 h whereas the same cultures showed low MI for each PHA stimulated culture. (author).

  1. DHT and IGF-1 in peripheral blood lymphocytes: new markers for the biological passport of athletes.

    Science.gov (United States)

    Mancini, A; Imperlini, E; Alfieri, A; Spaziani, S; Martone, D; Parisi, A; Orru, S; Buono, P

    2013-01-01

    We performed a pilot study using human peripheral blood lymphocytes (PBL) as a novel system to identify new biomarkers of dihydrotestosterone (DHT) and insulin-like growth factor-1 (IGF-1) abuse in sport. First, to obtain a gene signature, we treated cultures of lymphocytes from sedentary males with three doses of 0.237 microg/ml DHT, each of which is 80-fold the physiological concentration in young adult male serum, at days 0, 2 and 4, or with a single dose of 1.25 microg/ml IGF-1, which is 5-fold the physiological concentration in young adult male serum. We then used the Human Genome U133 Plus 2.0 microarray to identify a gene signature related to DHT or IGF-1 administration. Gene expression was evaluated after 7 and 21 days of DHT treatment, and after 24 h, 72 h and 7 days of IGF-1 treatment. Microarray analysis yielded a list of genes whose expression was altered after DHT or IGF-1 treatment. Among these we selected the genes that are most representative of the pathways associated with skeletal and muscular disorders using the IPA bioinformatics tool. We identified six (IDO1, CXCL13, CCL1, GZMB, VDR and IL2RA) and two (FN1 and RAB31) genes that were up-regulated in lymphocytes from sedentary subjects after 7 days of DHT and IGF-1 treatment, respectively. The expression of these genes in lymphocytes from differently trained athletes was either down-regulated or similar to that in lymphocytes from sedentary subjects. This finding suggests that up-regulation was due to the drug and not to physical exercise. In conclusion, we demonstrate that PBL can be useful in anti-doping checks, and we describe new biomarkers of DHT and IGF-1 abuse which can be included in the Athlete's Biological Passport.

  2. Effect of catecholamines and thermal exposure on lymphocyte proliferation, IL-1α & β in buffaloes

    Directory of Open Access Journals (Sweden)

    R.C. Upadhyay

    2010-02-01

    Full Text Available In order to study the effect of catecholamines (epinephrine/norepinephrine and thermal exposure on in vitro buffalo Lymphocyte Proliferation (LP apparently healthy 2- 2 1/2 years old Murrah buffalo heifers maintained as per the standard feeding and management practices were selected from Institute herd. Jugular blood was collected in the forenoon on the day of experiment and processed for Total Leucocyte Count (TLC and Differential Leucocyte Count (DLC. Lymphocyte proliferation assays were performed using whole blood and cells were incubated with epinephrine and norepinephrine (1, 1.5,2 ng/ml at 37oC with 5% CO2. Cells were counted after 72 hrs of incubation and Lymphocyte Proliferation Index (LPI was calculated. Thermal stress effect on the cultures was observed after exposure at 45oC for 4 hr after 72hrs of incubation. The cells were separated from media and media was used for analysis of IL-1α & 1β by ELISA kit. Lymphocyte proliferation Index decreased in responses to Epinephrine and Norepinephrine (P<0.01. Concentration of epinephrine and norepinephrine (1, 1.5,2 ng /ml had no distinguishable effect on LPI. IL-1α & IL-1β levels when compared with control in supernatant (exposed to 45°C were low (P<0.01 and P<0.05, respectively. There was a significant positive correlation between LPI and IL-1α (r=0.80; P<0.01 and between LPI and IL-1β (r=0.78; P<0.05. The study indicated that lymphocyte proliferation in vitro and IL-1α & β levels were affected by catecholamines and thermal exposure. Further the levels of catecholamines had significant (P<0.01 negative effect on LPI indicating that catecholamines levels modulate immunity through IL-1α and IL-1β in buffaloes.

  3. Preferential S phase entry and apoptosis of CD4(+) T lymphocytes of HIV-1-infected patients after in vitro cultivation.

    Science.gov (United States)

    Patki, A H; Zielske, S P; Sieg, S F; Lederman, M M

    2000-12-01

    We have studied the relationship between spontaneous apoptosis and cell cycle perturbations in circulating peripheral blood lymphocytes of HIV-1-infected patients and healthy controls. PBMC obtained from HIV-1-infected patients and healthy controls were incubated in culture medium for 48 h. Cells were separated into CD4(+) and CD8(+) populations using immunomagnetic beads. Apoptosis and cell cycle phases were measured by propidium iodide staining and bromodeoxyuridine (BrdU) incorporation followed by flow cytometric analyses. In experiments using cells obtained from HIV-1-infected patients, spontaneous apoptosis was more frequent in CD4(+) T lymphocytes than in CD8(+) T lymphocytes (17.6% vs 9.5%, P lymphocytes was comparable (4.5% vs 5.1%). Lymphocytes obtained from patients were more frequently in S phase than healthy controls' cells (2.2 +/- 0.9% vs 0.5 +/- 0.2%, P 0.4). Kinetic analyses using BrdU and PI staining revealed that S phase cells were more likely to become apoptotic than resting (G(0)-G(1)) cells (28.4% +/- 10.3% vs 11.3% +/- 9.9% in patients, P Lymphocytes obtained from HIV-1-infected persons are activated in vivo to enter S phase and to undergo spontaneous apoptosis after brief in vitro cultivation. The present studies indicate that most apoptotic cells in this system are CD4(+) and kinetic analyses reveal that S phase cells are more likely to undergo spontaneous apoptosis than G(0)-G(1) cells. Accelerated cell death in HIV-1 disease may contribute to the failure of lymphocyte responsiveness to appropriate T cell receptor stimulation.

  4. Alteration of peripheral blood lymphocyte subsets in acute pancreatitis

    Institute of Scientific and Technical Information of China (English)

    Miroslawa Pietruczuk; Milena I Dabrowska; Urszula Wereszczynska-Siemiatkowska; Andrzej Dabrowski

    2006-01-01

    AIM: To evaluate peripheral blood lymphocyte subsets in patients with acute pancreatitis (AP).METHODS: Twenty patients with mild AP (M-AP) and 15 with severe AP (S-AP) were included in our study. Peripheral blood lymphocytes were examined at d 1-3, 5,10 and 30 by means of flow cytometry.RESULTS: A significant depletion of circulating lymphocytes was found in AP. In the early AP, the magnitude of depletion was similar for T- and B- lymphocytes. In the late course of S-AP, B-lymphocytes were much more depleted than T-lymphocytes. At d 10, strong shift in the CD7+/CD19+ ratio implicating predominance of Tover B-lymphocytes in S-AP was found. Among T-lymphocytes, the significant depletion of the CD4+ population was observed in M-AP and S-AP, while CD8+ cells were in the normal range. Lymphocytes were found to strongly express activation markers: CD69, CD25, CD28,CD38 and CD122. Serum interleukin-2 (IL-2), IL-4, IL-5,IL-10, interferon-γ (IFN-γ) and tumor necrosis factor-α(TNF-α) levels were significantly increased in both forms of AP. The magnitude of elevation of cytokines known to be produced by Th2 was much higher than cytokines produced by Th1 cells.CONCLUSION: AP in humans is characterized by significant reduction of peripheral blood T- and B-lymphocytes.

  5. Natural and artefactual aneuploidy in human lymphocytes in extreme old age.

    Science.gov (United States)

    Lezhava, T; Khmaladze, E; Lokhadze, T; Buadze, T

    2008-04-01

    At present it is generally believed that aging is accompanied by an increase in the numbers of cells with altered chromosome sets. However, the problem of aneuploidy in extreme old age (80 years and over) has hardly been subjected to systematic investigation. The purpose of this study was to examine quantitative chromosome changes and relationship between "artefactual" and "natural" aneuploidy in subjects from 80 to 114 years of age using karyotype analyses. Chromosomes were studied in 1136 karyotypes of relatively round metaphases from 40 lymphocyte cultures obtained from 40 apparently normal subjects aged from 80 to 114 ( including 26 men and 14 women). 964 karyotypes from 48 donors aged 20 to 48 served as control. Lymphocytes were cultivation by modification method (Lezhava, 2006). It is suggested that the probability of natural aneuploidy rises substantially among elderly women; while that of artefactual aneuploidy is increased among elderly men; the problem of natural aneuploidy in males remains unclear.

  6. Influence of cosmic radiationon lymphocyte micronucleus,serum lipid peroxide and antioxidation capacity inaircrew members

    Institute of Scientific and Technical Information of China (English)

    2002-01-01

    This study looks into the influence of cosmic radiation at high altitudes on human bodies. Results reveal that the cytokinesis-block micronuclei (CBMN) and conventional cultured micronuclei in peripheral blood lymphocytes, serum levels of lipid peroxide, superoxide dismutase, and the total antioxidation capacity by chemical colorimetry all increased significantly in aircrew members. There exists a linear relationship between the CBMN and the average annual effective doses of radiation received or the average annual flying hours. With both of them, a trend shows that the serum lipid peroxide levels increase as well. Either the lipid peroxide or CBMN can sensitively reflect the recent changes in flight load. These findings indicate that cosmic radiation impairs the stability of chromosomes and genome, and induces lipid oxidative damage in aircrews; Lymphocyte CBMN and serum lipid peroxide can be used as monitoring indicators in the cosmic radiation protection for aircrew members.

  7. Clinical scale rapid expansion of lymphocytes for adoptive cell transfer therapy in the WAVE® bioreactor

    Science.gov (United States)

    2012-01-01

    Background To simplify clinical scale lymphocyte expansions, we investigated the use of the WAVE®, a closed system bioreactor that utilizes active perfusion to generate high cell numbers in minimal volumes. Methods We have developed an optimized rapid expansion protocol for the WAVE bioreactor that produces clinically relevant numbers of cells for our adoptive cell transfer clinical protocols. Results TIL and genetically modified PBL were rapidly expanded to clinically relevant scales in both static bags and the WAVE bioreactor. Both bioreactors produced comparable numbers of cells; however the cultures generated in the WAVE bioreactor had a higher percentage of CD4+ cells and had a less activated phenotype. Conclusions The WAVE bioreactor simplifies the process of rapidly expanding tumor reactive lymphocytes under GMP conditions, and provides an alternate approach to cell generation for ACT protocols. PMID:22475724

  8. Clinical scale rapid expansion of lymphocytes for adoptive cell transfer therapy in the WAVE® bioreactor

    Directory of Open Access Journals (Sweden)

    Somerville Robert PT

    2012-04-01

    Full Text Available Abstract Background To simplify clinical scale lymphocyte expansions, we investigated the use of the WAVE®, a closed system bioreactor that utilizes active perfusion to generate high cell numbers in minimal volumes. Methods We have developed an optimized rapid expansion protocol for the WAVE bioreactor that produces clinically relevant numbers of cells for our adoptive cell transfer clinical protocols. Results TIL and genetically modified PBL were rapidly expanded to clinically relevant scales in both static bags and the WAVE bioreactor. Both bioreactors produced comparable numbers of cells; however the cultures generated in the WAVE bioreactor had a higher percentage of CD4+ cells and had a less activated phenotype. Conclusions The WAVE bioreactor simplifies the process of rapidly expanding tumor reactive lymphocytes under GMP conditions, and provides an alternate approach to cell generation for ACT protocols.

  9. Generation of tumor-targeted human T lymphocytes from induced pluripotent stem cells for cancer therapy.

    Science.gov (United States)

    Themeli, Maria; Kloss, Christopher C; Ciriello, Giovanni; Fedorov, Victor D; Perna, Fabiana; Gonen, Mithat; Sadelain, Michel

    2013-10-01

    Progress in adoptive T-cell therapy for cancer and infectious diseases is hampered by the lack of readily available, antigen-specific, human T lymphocytes. Pluripotent stem cells could provide an unlimited source of T lymphocytes, but the therapeutic potential of human pluripotent stem cell-derived lymphoid cells generated to date remains uncertain. Here we combine induced pluripotent stem cell (iPSC) and chimeric antigen receptor (CAR) technologies to generate human T cells targeted to CD19, an antigen expressed by malignant B cells, in tissue culture. These iPSC-derived, CAR-expressing T cells display a phenotype resembling that of innate γδ T cells. Similar to CAR-transduced, peripheral blood γδ T cells, the iPSC-derived T cells potently inhibit tumor growth in a xenograft model. This approach of generating therapeutic human T cells 'in the dish' may be useful for cancer immunotherapy and other medical applications.

  10. Exposure of human nasal epithelial cells to formaldehyde does not lead to DNA damage in lymphocytes after co-cultivation.

    Science.gov (United States)

    Neuss, Simone; Moepps, Barbara; Speit, Günter

    2010-07-01

    We performed in vitro co-cultivation experiments with primary human nasal epithelial cells (HNEC) and isolated lymphocytes to investigate whether reactive formaldehyde (FA) can be passed on from nasal epithelial cells (site of first contact) to lymphocytes located in close proximity and induce DNA damage in these cells. A modified comet assay was used as a sensitive method for the detection of FA-induced DNA-protein cross links (DPX) because DPX are the most relevant type of FA-induced DNA damage. Our results clearly indicate that co-cultivation of lymphocytes with HNEC exposed to FA for 1 h causes a concentration-related induction of DPX in lymphocytes when co-cultivation takes place in the exposure medium. However, when the exposure medium is changed after FA treatment of HNEC and before lymphocytes are added, no induction of DPX is measured in lymphocytes even after exposure of HNEC to high FA concentrations (300 microM) and extended co-cultivation (4 h). Direct measurement of FA in the cell culture medium by a sensitive fluorescent detection kit indicated that FA is actually not released even from highly exposed cells into the cell culture medium. These results suggest that FA that has entered nasal epithelial cells is not released and does not damage other cells in close proximity to the epithelial cells. If these results also apply to the in vivo situation, FA would only be genotoxic towards directly exposed cells (site of first contact) and there should be no significant delivery of inhaled FA to other cells and distant sites. Our results do not support a recently proposed hypothetic mechanism for FA-induced leukaemia by damaging circulating haematopoietic stem cells or haematopoietic progenitor cells in nasal passages, which then travel to the bone marrow and become initiated leukaemic stem cells.

  11. Idelalisib for the treatment of chronic lymphocytic leukemia/small lymphocytic lymphoma.

    Science.gov (United States)

    Barrientos, Jacqueline C

    2016-09-01

    Idelalisib is a first-in-class selective oral PI3Kδ inhibitor for the treatment of patients with relapsed chronic lymphocytic leukemia (CLL) and small lymphocytic lymphoma, a predominantly elderly population with high comorbidity. The drug promotes apoptosis in primary CLL cells ex vivo, independent of common prognostic markers and inhibits CLL cell homing, migration and adhesion to cells in the microenvironment. Idelalisib has shown efficacy with acceptable safety as monotherapy and combination therapy in relapsed/refractory CLL. Idelalisib has clinical activity in patients with CLL with del(17p). The development of other novel B-cell-targeted agents provides the opportunity to evaluate additional idelalisib treatment combinations for their potential to further improve outcomes in CLL/small lymphocytic lymphoma.

  12. HLA-DP and bonemarrow transplantation: DP-incompatibility and severe acute graft versus host disease

    DEFF Research Database (Denmark)

    Ødum, Niels; Platz, P; Jakobsen, B K

    1987-01-01

    of the various HLA class II antigens was observed between the groups. Similarly, no significant differences in stimulatory capability in secondary mixed lymphocyte culture (MLC) were seen. The distribution of T helper/inducer (CD4+), T suppressor/cytotoxic (CD8+), and NK cells was similar in active JCA, JCA...

  13. Rapid diagnosis of vitamin D-dependent rickets type II by use of phytohemagglutinin-stimulated lymphocytes.

    Science.gov (United States)

    Takeda, E; Kuroda, Y; Saijo, T; Toshima, K; Naito, E; Kobashi, H; Iwakuni, Y; Miyao, M

    1986-03-28

    The interactions of 1,25-dihydroxyvitamin D3 [1,25-(OH)2D3] with phytohemagglutinin (PHA)-stimulated lymphocytes from normal subjects and three patients with vitamin D-dependent rickets (DDR) type II were investigated. Impaired nuclear uptake and normal cytosol binding of [3H]1,25-(OH)2D3 were observed with PHA-stimulated lymphocytes of these patients as with their cultured skin fibroblasts. Furthermore, the incorporation of [14C]thymidine into PHA-stimulated lymphocytes of the patients was not reduced by 1,25-(OH)2D3, which is known to inhibit proliferation of various cells. These findings suggest that 1,25-(OH)2D3 receptors are reduced or absent in patients with DDR type II. Thus, the capacities of cytosol binding and nuclear uptake of 1,25-(OH)2D3 in PHA-stimulated lymphocytes seem to reflect those of endo-organs such as the intestine and bone. These findings show that a test of the effect of 1,25-(OH)2D3 on thymidine incorporation into PHA-stimulated lymphocytes is useful for rapid diagnosis of DDR type II.

  14. Single arc volumetric-modulated arc therapy is sufficient for nasopharyngeal carcinoma: a dosimetric comparison with dual arc VMAT and dynamic MLC and step-and-shoot intensity-modulated radiotherapy

    OpenAIRE

    NING, ZHONG-HUA; Mu, Jin-Ming; Jin, Jian-Xue; Li, Xiao-Dong; LI, QI-LIN; GU, WEN-DONG; Huang, Jin; Han, Yang; PEI, HONG-LEI

    2013-01-01

    Background The performance of single arc VMAT (VMAT1) for nasopharyngeal carcinoma (NPC) on the Axesse linac has not been well described in previous studies. The purpose of this study is to assess the feasibility of VMAT1 for NPC by comparing the dosimetry, delivery efficiency, and accuracy with dual arc VMAT (VMAT2), dynamic MLC intensity-modulated radiotherapy (dIMRT), and step-and-shoot intensity-modulated radiotherapy (ssIMRT). Methods Twenty consecutive patients with non-metastatic NPC w...

  15. Induction of apoptosis in murione spleen lymphocytes using carbon ion beam; Induction de l'apoptose dans les lymphocytes spleniques de souris par un faisceau d'ions carbone

    Energy Technology Data Exchange (ETDEWEB)

    Holl, V.; Coelho, D. [Lab. de Cancerologie Experimentale et de Radiobiologie, LCER, Inst. de Recherche contre les Cancers de l' Appareil Digestif, Hopitaux Universitaires, Strasbourg (France); Weltin, D. [Faculte de Medecine, Strasbourg (France); Dufour, P. [Lab. de Cancerologie Experimentale et de Radiobiologie, LCER, Inst. de Recherche contre les Cancers de l' Appareil Digestif, Hopitaux Universitaires, Strasbourg (France); Denis, J.M. [Faculte de Medecine, Univ. Catholique de Louvain, Bruxelles (Belgium); Florentin, I. [Centre National de la Recherche Scientifique, CNRS, Unite de Recherche Associee, Hopital Cochin, Paris (France); Mathieu, J. [Centre de Recherche du Service de Sante des Armees, CRSSA, La Tronche (France); Gueulette, J. [Faculte de Medecine, Univ. Catholique de Louvain, Bruxelles (Belgium); Bischoff, P. [Lab. de Cancerologie Experimentale et de Radiobiologie, LCER, Inst. de Recherche contre les Cancers de l' Appareil Digestif, Hopitaux Universitaires, Strasbourg (France)

    2001-02-01

    To assess the capacity of heavy ions to induce apoptosis in lymphocytes, mice have been irradiated with accelerated carbon ions (95 MeV/nucleon) at doses ranging from 0.1 to 4 Gy. Their spleens were removed 24 h later and gently dissociated to prepare a single cell suspension. Mononuclear cells were then maintained in culture at 37{sup o}C, and the occurrence of apoptosis in these cells was analysed 24 h later. Lymphocytes were also irradiated in vitro, in the presence of Ac-DEVD-CHO, a potent caspase-3 and -7 inhibitor. Results from three experiments performed at the Grand Accelerateur National d'Ions Lourds (GANIL, Caen, France) are reported here. They indicate that carbon ions induce a marked, dose-dependent, reduction of the spleen weight and cellularity. However, in sharp contrast with spleen cells prepared from X-ray irradiated mice, only a slight increase of apoptosis is evidenced in cultured lymphocytes from mice irradiated with heavy ions. The significance of such results is discussed. So far, few data exist concerning the biological effects of heavy ions, in particular their capacity to induce apoptosis in lymphocytes; the present study provides useful clues for further investigations. (author)

  16. The association between chronic lymphocytic thyroiditis and thyroid tumors.

    Science.gov (United States)

    Tamimi, Dalal M

    2002-04-01

    An association between lymphocytic thyroiditis and thyroid papillary carcinoma is still controversial. To assess the relationship, a histopathologic analysis of surgically resected thyroid tumors together with the frequency and severity of chronic lymphocytic infiltration of the thyroid among patients with follicular adenoma, follicular carcinoma, and papillary carcinoma was performed. The prevalence of lymphocytic infiltrate, which is indicative of autoimmune thyroiditis, was significantly higher in patients with papillary carcinoma (58%) than in patients with follicular carcinoma (20%) or follicular adenoma (14%). The lymphocytic infiltration within the tumor compared with the severity of thyroiditis in the nontumorous tissue. Therefore, the association between chronic lymphocytic thyroiditis and papillary carcinoma was confirmed. The possibility that an immunologic mechanism involved in the pathogenesis of papillary carcinoma stimulates lymphocytic infiltration in the thyroid tissue through an autoimmune mechanism is suggested.

  17. Two structurally identical mannose-specific jacalin-related lectins display different effects on human T lymphocyte activation and cell death.

    Science.gov (United States)

    Benoist, Hervé; Culerrier, Raphaël; Poiroux, Guillaume; Ségui, Bruno; Jauneau, Alain; Van Damme, Els J M; Peumans, Willy J; Barre, Annick; Rougé, Pierre

    2009-07-01

    Plant lectins displaying similar single sugar-binding specificity and identical molecular structure might present various biological effects. To explore this possibility, the effects on human lymphocytes of two mannose-specific and structurally closely related lectins, Morniga M from Morus nigra and artocarpin from Artocarpus integrifolia were investigated. In silico analysis revealed that Morniga M presents a more largely open carbohydrate-binding cavity than artocarpin, probably allowing interactions with a broader spectrum of carbohydrate moieties. In vitro, Morniga M interacted strongly with the lymphocyte surface and was uptaken quickly by cells. Morniga M and artocarpin triggered the proliferation and activation of human T and NK lymphocytes. A minority of B lymphocytes was activated in artocarpin-treated culture, whereas Morniga M favored the emergence of CD4+ CD8+ T lymphocytes. Moreover, cell death occurred in activated PBMC, activated T lymphocytes, and Jurkat T leukemia cells incubated with Morniga M only. The biological effects of both lectins were dependent on carbohydrate recognition. The Morniga M-induced cell death resulted, at least in part, from caspase-dependent apoptosis and FADD-dependent receptor-mediated cell death. Finally, Morniga M, but not artocarpin, triggered AICD of T lymphocytes. In conclusion, both lectins trigger lymphocyte activation, but only Morniga M induces cell death. In spite of similar in vitro mannose-binding specificities and virtually identical structure, only Morniga M probably interacts with carbohydrate moieties bound to molecules able to induce cell death. The present data suggest that subtle alterations in N-glycans can distinguish activation and cell death molecules at the lymphocyte surface.

  18. Toxicological effects of ultraviolet radiation on lymphocyte cells with different manganese superoxide dismutase Ala16Val polymorphism genotypes.

    Science.gov (United States)

    dos Santos Montagner, Greice Franciele Feyh; Sagrillo, Michele; Machado, Michel Mansur; Almeida, Renata Chequeller; Mostardeiro, Clarice Pinheiro; Duarte, Marta Maria Medeiros Frescura; da Cruz, Ivana Beatrice Mânica

    2010-08-01

    The aim of this study was to investigate whether there is a differential response of lymphocytes from healthy MnSOD genotype subjects to oxidative stress. We used UV radiation as a toxic agent due to its genotoxic effects associated with chromosome aberrations caused by breaks in the DNA strands. Cellular growth rate, cell viability, mitotic index, chromosomal instability and biomarkers of oxidative metabolism were analysed in lymphocyte cells from healthy adults with different Ala16Val MnSOD polymorphisms that produce tree genotypes: AA, VV and AV. We found a differential response to UV exposure in cultures of lymphocyte cells from Ala16Val genotype donors. In general, AA cell cultures presented higher viability and mitotic index and lower TBARS levels than VV and AV cells for both the control and UV exposure groups. However, when we compared the DNA damage among the three genotypes, AA lymphocyte cells presented the highest damage from UV exposure. These data suggest that the Ala16Val polymorphism affects the response of cellular oxidative metabolism in different ways.

  19. Development of a serum-free medium for in vitro expansion of human cytotoxic T lymphocytes using a statistical design

    Directory of Open Access Journals (Sweden)

    Lee Gyun

    2010-09-01

    Full Text Available Abstract Background Serum-containing medium (SCM, which has a number of poorly defined components with varying concentrations, hampers standardization of lymphocyte cultures. In order to develop a serum-free medium (SFM for the expansion of human lymphocytes from peripheral blood mononuclear cells (PBMCs, a statistical optimization approach based on a fractional factorial method and a response surface method was adopted. A basal medium was prepared by supplementing RPMI1640 medium with insulin, albumin, ferric citrate, ethanolamine, fatty acids, glutamine, sodium pyruvate, 2-mercaptoethanol, 1-thioglycerol, nonessential amino acids, and vitamins. We identified additional positive determinants and their optimal concentrations for cell growth through a statistical analysis. Results From a statistical analysis using the fractional factorial method, cholesterol and polyamine supplement were identified as positive determinants for cell growth. Their optimal concentrations were determined by the response surface method. The maximum viable cell concentration in the developed SFM was enhanced by more than 1.5-fold when compared to that in RPMI1640 supplemented with 10% fetal bovine serum (FBS. Furthermore, a cytotoxicity assay and an enzyme-linked immunospot assay revealed that the effector function of cytotoxic T lymphocytes generated from PBMCs grown in SFM, by stimulation of peptide-presenting dendritic cells, was retained or even better than that in SCM. Conclusions The use of a developed SFM with cholesterol and polyamine supplement for human lymphocyte culture resulted in better growth without loss of cellular function when compared to SCM.

  20. Cell-mediated immune response of synovial fluid lymphocytes to ureaplasma antigen in Reiter's syndrome

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    Pavlica Ljiljana

    2003-01-01

    Full Text Available INTRODUCTION Reiter's syndrome (RS is an seronegative arthritis that occurs after urogenital or enteric infection which in addition with occular and/or mucocutaneous manifestations presents complete form of disease. According to previous understanding arthritis in the RS is the reactive one, which means that it is impossible to isolate its causative agent. However, there are the more and more authors suggesting that arthritis in the urogenital form of disease is caused by the infective agent in the affected joint. This suggestion is based on numerous studies on the presence of Chlmaydia trachomatis and Ureaplasma urealyticum in the inflamed joint by using new diagnostic methods in molecular biology published in the recent literature [1-3]. Besides, numerous studies of the humoral and cell-mediated immune response to "triggering" bacteria in the affected joint have supported previous suggestions [4-7]. Aim of the study was to determine whether synovial fluid T-cells specifically recognize the "triggering" bacteria presumably responsible for the Reiter's syndrome. METHOD The 3H-thymidine uptake procedure for measuring lymphocyte responses was applied to lymphocytes derived concurrently from synovial fluid (SF and from peripheral blood (PB [8]. Ureaplasma antigen and mitogen PHA stimulated lymphocytes in 24 RS patients (24 PB samples, 9 SF samples and the results were compared with those found in 10 patients with rheumatoid arthritis (RA (10 PB samples, 5 SF samples. Preparation of ureaplasma antigen. Ureaplasma was cultured on cell-free liquid medium [9]. Sample of 8 ml was heat-inactivated for 15 minutes at 601C and permanently stirred with magnetic mixer. The sample was centrifuged at 2000 x g for 40 minutes and than deposits carefully carried to other sterile glass tubes (Corex and recentrifuged at 9000 x g for 30 minutes. The deposit was washed 3 times in sterile 0.9% NaCl, and final sediment was resuspended in 1.2 ml sterile 0.9% Na

  1. Chronic lymphocytic leukemia: case-based session.

    Science.gov (United States)

    Rai, K R; Döhner, H; Keating, M J; Montserrat, E

    2001-01-01

    Drs. Hartmut Döhner, Michael J. Keating, Kanti R. Rai and Emili Montserrat form the panel to review chronic lymphocytic leukemia (CLL) while focusing on the clinical features of a particular patient. The pace of progress in CLL has accelerated in the past decade. The pathophysiological nature of this disease, as had been known in the past, was based largely on the intuitive and empiric notions of two leaders in hematology, William Dameshek and David Galton. Now the works of a new generation of leaders are providing us with the scientific explanations of why CLL is a heterogeneous disease, perhaps consisting of at least two separate entities. In one form of CLL, the leukemic lymphocytes have a surface immunoglobulin (Ig) variable region gene that has undergone somatic mutations, with tell-tale markers suggesting that these cells had previously traversed the germinal centers. Such patients have a distinctly superior prognosis than their counterparts whose leukemic lymphocytes IgV genes have no mutations (these are indeed immunologically naive cells), who have a worse prognosis. The introduction of fluorescence in situ hybridization (FISH) technique has provided us with new insights into the diverse chromosomal abnormalities that can occur in CLL, and which have significant impact on the clinical behavior and prognosis of patients with this disease. Major advances in therapeutics of CLL also have occurred during the past decade. Two monoclonal antibodies, Campath-1H (anti-CD52) and rituximab (anti-CD20), and one nucleoside analogue, fludarabine, have emerged as three agents of most promise in the front-line treatment of this disease. Studies currently in progress reflect our attempts to find the most effective manner of combining these agents to improve the overall survival statistics for CLL patients. As in many other hematological malignancies, high dose chemotherapy followed by autologous or HLA-compatible allogeneic stem cells rescue strategies are under study as

  2. Natural History Study of Monoclonal B Cell Lymphocytosis (MBL), Chronic Lymphocytic Leukemia/Small Lymphocytic Lymphoma (CLL/SLL), Lymphoplasmacytic Lymphoma (LPL)/Waldenstrom Macroglobulinemia (WM), and Splenic Marginal Zone Lymphoma (SMZL)

    Science.gov (United States)

    2017-10-05

    B-Cell Chronic Lymphocytic Leukemia; Monoclonal B-Cell Lymphocytosis; Lymhoma, Small Lymphocytic; Chronic Lymphocytic Leukemia; Lymphoplasmacytic Lymphoma; Waldenstrom Macroglobulinemia; Splenic Marginal Zone Lymphoma

  3. Phenotypic and Functional Analysis of Porcine T Lymphocytes

    Institute of Scientific and Technical Information of China (English)

    李华; 陈应华

    2001-01-01

    Porcine and other higher mammals express clusters of differentiation (CD) antigens on the surface of T lymphocytes, such as CD2, CD3, CD4, CD8, etc. However, in porcine, a high percentage of the CD4+ CD8-T lymphocyte subpopulation exist in the peripheral blood and the ratio of the CD4+ and CD8+ T lymphocyte subpopulations is reversed. These differences bring new challenges to better understanding of the phenotype and function of porcine T lymphocytes in antigen recognition and immune response.

  4. Recent advances in chronic lymphocytic leukemia

    Directory of Open Access Journals (Sweden)

    N Vyas

    2012-01-01

    Full Text Available Chronic lymphocytic leukemia (CLL was largely considered to be a disease of slow progression, standard treatment with Chlorambucil and having almost similar prognosis. With the introduction of molecular methods for understanding the disease pathophysiology in CLL there has been a remarkable change in the approach towards the disease. The variation in B-cell receptor response and immunoglobulin heavy chain variable region (IGHV mutation, genetic aberration and defect in apoptosis and proliferation has had an impact on therapy initiation and prognosis. Early diagnosis of molecular variant is therefore necessary in CLL.

  5. Chronic pain: cytokines, lymphocytes and chemokines.

    Science.gov (United States)

    de Miguel, Marcia; Kraychete, Durval Campos; Meyer Nascimento, Roberto Jose

    2014-01-01

    Chronic pain is a debilitating condition and, in most cases, difficult to treat. A prominent example of this is neuropathic pain. Understanding pathophysiological mechanisms of pain and, therefore, making this knowledge into an effective treatment is still a challenge to experts. Pain can now be considered as a neuro-immune disorder, since recent data indicate critical involvement of innate and adaptive immune responses following injury, and this interaction plays an important role in the onset and perpetuation of chronic pain. The aim of this article is to review the relationship between immune system and chronic pain, especially about neuropathic pain, and focusing on cytokines, chemokines and lymphocytes.

  6. Spinal epidural compression in chronic lymphocytic leukemia.

    Science.gov (United States)

    Michalevicz, R; Burstein, A; Razon, N; Reider, I; Ilie, B

    1989-11-01

    Spinal epidural compression is a rare neurologic complication in patients with lymphoma. It occurs mostly in those with intermediate-grade to high-grade malignancy disease. This type of neurologic involvement has not been described in chronic lymphocytic leukemia (CLL). A patient with a long, stable CLL course developed spinal epidural compression and consequently died. The frequency of spinal epidural compression in lymphoma, according to the histologic subtypes and the considerations in making the right choice of therapy are discussed in light of the presented case.

  7. HLA-DP related suppression of mixed lymphocyte reaction with alloactivated lymphocytes

    DEFF Research Database (Denmark)

    Ødum, Niels; Hofmann, B; Jakobsen, B K

    1986-01-01

    We studied the influence of HLA class I and class II antigens on the suppression of the MLR induced by primed lymphocytes (PLs) alloactivated in vitro. The suppression of 14 different PLs of 83 MLRs was analyzed. The PLs were primed against (i) HLA-DP (SB) (ii) HLA-DR/DQ or (iii) both HLA-DP and DR...

  8. Characterization of the lymphocyte substance P receptor

    Energy Technology Data Exchange (ETDEWEB)

    McGillis, J.P.; Organist, M.L.; Payan, D.G.

    1986-03-01

    Recent studies have provided evidence that tissues of the immune and reticuloendothelial systems are influenced by various hormones and neuropeptides. While the interrelationship between these peptides and the immune system, and the physiological relevance of their effects is not clear, a variety of highly specific, receptor-mediated effects has been demonstrated. One neuropeptide for which immunomodulatory effects have been identified is substance P (SP). SP acts as a potent T-cell mitogen, and enhances the mitogenic effects of PHA. Radioreceptor and FACS binding studies suggest that the SP receptor is present on a discrete population of T-cells and has a Kd of 0.87 nM and a density of 24,000. Studies have been initiated to biochemically characterize the lymphocyte SP receptor on IM-9 lymphoblasts. (/sup 125/I)-labeled SP was covalently crosslinked to the receptor using disuccinimidyl suberate and solubilized. Two radioactive bands of m.w. 55,000 and 33,000 were seen when the solubilized crosslinked receptor was electrophoresed on SDS gels. The authors are currently using a combination of HPLC immunoaffinity and reverse-phase chromatography to purify the receptor. The initial studies indicate that the receptor can be purified to homogeneity using this approach. The biochemical characterization of this receptor should provide a better understanding of its importance not only on lymphocytes, but in other tissues as well.

  9. Spondylarthritis in the absence of B lymphocytes.

    Science.gov (United States)

    Baeten, Dominique; Kruithof, Elli; Breban, Maxime; Tak, Paul P

    2008-03-01

    The highly effective treatment of rheumatoid arthritis by B cell depletion and the presence of B cells in the peripheral and axial lesions of patients with spondylarthritis (SpA) raise the question as to whether B lymphocytes could also be an appropriate therapeutic target in the latter disease. We describe 2 male HLA-B27-positive patients who had active SpA despite absence of B cells. One patient developed SpA with sacroiliitis and asymmetric oligoarthritis after having been diagnosed as having severe Bruton agammaglobulinemia. Since extensive investigations excluded an infectious origin of the SpA, this case illustrates that functional B cells and/or gamma globulins are not strictly required for SpA pathogenesis. The second patient had severe axial and peripheral SpA that was treated successfully with etanercept. After discontinuation of etanercept treatment because of non-Hodgkin's B cell lymphoma, both axial and peripheral SpA symptoms relapsed rapidly, and this exacerbation of articular disease activity was not modulated by successful B cell depletion therapy for the lymphoma. Although case reports have obvious limitations, our clinical observations provide evidence that active SpA can occur in the absence of functional mature B cells and thus emphasize the need for systematic studies of the exact role and function of B lymphocytes in this disease.

  10. Secondary autoimmune cytopenias in chronic lymphocytic leukemia.

    Science.gov (United States)

    Rogers, Kerry A; Woyach, Jennifer A

    2016-04-01

    Secondary autoimmune cytopenias in chronic lymphocytic leukemia are distinct clinical entities that require specific management. These autoimmune disorders have a complex pathogenesis that involves both the leukemic cells and the immune environment in which they exist. The mechanism is not the same in all cases, and to varying degrees involves the chronic lymphocytic leukemia (CLL) cells in antibody production, antigen presentation, and stimulation of T cells and bystander polyclonal B cells. Diagnosis of autoimmune cytopenias can be challenging as it is difficult to differentiate between autoimmunity and bone marrow failure due to disease progression. There is a need to distinguish these causes, as prognosis and treatment are not the same. Evidence regarding treatment of secondary autoimmune cytopenias is limited, but many effective options exist and treatment can be selected with severity of disease and patient factors in mind. With new agents to treat CLL coming into widespread clinical use, it will be important to understand how these will change the natural history and treatment of autoimmune cytopenias.

  11. TACI Expression and Signaling in Chronic Lymphocytic Leukemia

    Science.gov (United States)

    Mamara, Antigoni; Germenis, Anastasios E.; Kompoti, Maria; Palassopoulou, Maria; Mandala, Eudokia; Banti, Anastasia; Giannakoulas, Nikolaos

    2015-01-01

    TACI is a membrane receptor of BAFF and APRIL, contributing to the differentiation and survival of normal B cells. Although malignant B cells are also subjected on TACI signaling, there is a remarkable intradisease and interindividual variability of TACI expression in B-cell malignancies. The aim of our study was to explore the possible role of TACI signaling in the biology of chronic lymphocytic leukemia (CLL), including its phenotypic and clinical characteristics and prognosis. Ninety-four patients and 19 healthy controls were studied. CLL patients exhibited variable TACI expression, with the majority of cases displaying low to undetectable TACI, along with low to undetectable BAFF and increased APRIL serum levels compared to healthy controls. CLL cells with high TACI expression displayed a better survival capacity in vitro, when cultured with BAFF and/or APRIL. Moreover, TACI expression was positively correlated with the presence of monoclonal gammopathy and inversely with CD11c expression. Therefore, our study provides further evidence for the contribution of BAFF/APRIL signaling to CLL biology, suggesting also that TACI detection might be useful in the selection of patients for novel targeting therapeutic approaches. PMID:25950010

  12. TACI Expression and Signaling in Chronic Lymphocytic Leukemia

    Directory of Open Access Journals (Sweden)

    Antigoni Mamara

    2015-01-01

    Full Text Available TACI is a membrane receptor of BAFF and APRIL, contributing to the differentiation and survival of normal B cells. Although malignant B cells are also subjected on TACI signaling, there is a remarkable intradisease and interindividual variability of TACI expression in B-cell malignancies. The aim of our study was to explore the possible role of TACI signaling in the biology of chronic lymphocytic leukemia (CLL, including its phenotypic and clinical characteristics and prognosis. Ninety-four patients and 19 healthy controls were studied. CLL patients exhibited variable TACI expression, with the majority of cases displaying low to undetectable TACI, along with low to undetectable BAFF and increased APRIL serum levels compared to healthy controls. CLL cells with high TACI expression displayed a better survival capacity in vitro, when cultured with BAFF and/or APRIL. Moreover, TACI expression was positively correlated with the presence of monoclonal gammopathy and inversely with CD11c expression. Therefore, our study provides further evidence for the contribution of BAFF/APRIL signaling to CLL biology, suggesting also that TACI detection might be useful in the selection of patients for novel targeting therapeutic approaches.

  13. T lymphocytes and NK cells in X-linked agammaglobulinemia.

    Science.gov (United States)

    Pituch-Noworolska, Anna; Zwonarz, Katarzyna; Błaut-Szlósarczyk, Anita; Szaflarska, Anna; Kowalczyk, Danuta; Siedlar, Maciej

    2013-01-01

    Seven boys with diagnosis of X-linked agammaglobulinemia on regular substitution of immunoglobulins were included into study. The patients showed episodes of infections but the clinical course was mild with good response to antibiotics. All patients developed, with time, the chronic sinusitis with proliferation of mucous membrane, two patients showed bronchiectases. The number of T lymphocytes, ratio of CD4:CD8 subpopulations, response to stimulation and NK number were assayed with flow cytometry and cell culture. Results showed CD4:CD8 ratio within normal value in majority of patients, reverse ratio in 2 boys, increased number of activated T cells (CD3/HLA-DR) in one of them. The number of NK cells was different from lack of these cells to high number. Response of T cells to stimulation (mitogens and CD3) were normal in majority of assays. There were no associations between clinical course and observed changes in T or NK cell populations. Further studies on number and function of NK cells are needed.

  14. Cytotoxic T lymphocyte activity in children infected with HIV.

    Science.gov (United States)

    Froebel, K S; Aldhous, M C; Mok, J Y; Hayley, J; Arnott, M; Peutherer, J F

    1994-01-01

    Of the Edinburgh cohort of approximately 130 children born to HIV-infected women, 9 are infected and alive. This article describes results from the first 18 months of a natural history study of seven of these, and two adopted children, studying the CD8 T cell-mediated cytotoxicity against HIV proteins (Gag, Tat, Pol, and Env), over time, and relating it to clinical progression and viral activity. Autologous EBV cell lines infected with vaccinia-HIV constructs were used as target cells, and bulk-cultured peripheral blood mononuclear cells as effector cells. The children ranged in age from 0 to 93 months, with six of the nine showing CTL activity to one or more HIV proteins. The specificity of the response was directed against Tat in the younger children, switching to Pol, then Gag or Env. Preliminary analysis of virological data showed no association between CTL and virus activity. The children with CTLs tended to be well clinically, but the cohort needs to be studied longer before conclusions can be made about CTL activity and HIV disease progression. Cytotoxic T lymphocyte activity has also been observed in two children diagnosed as HIV uninfected. These results show the importance of looking at CTL specificity, and may have implications in vaccine design.

  15. Ex vivo expansion of tumor-infiltrating lymphocytes from nasopharyngeal carcinoma patients for adoptive immunotherapy

    Institute of Scientific and Technical Information of China (English)

    Jia He; Xiao-Feng Tang; Qiu-Yan Chen; Hai-Qiang Mai; Zhou-Feng Huang; Jiang Li; Yi-Xin Zeng

    2012-01-01

    Establishing Epstein-Barr virus (EBV)-specific cytolytic T lymphocytes (EBV-CTLs) from peripheral blood mononuclear cells (PBMCs) for adoptive immunotherapy has been reported in EBV-associated malignancies including Hodgkin's lymphoma and nasopharyngeal carcinoma (NPC).In the current study,we performed ex vivo expansion of tumor-infiltrating lymphocytes (TILs) obtained from NPC biopsy specimens with a rapid expansion protocol using anti-CD3 monoclonal antibody (OKT3),recombinanthuman interleukin (IL)-2,and irradiated PBMCs from healthy donors to initiate the growth of TILs.Young TIL cultures comprised of more than 90% of CD3+ T cells,a variable percentage of CD3+CD8+ and CD3+CD4+ T cells,and less than 10% of CD3-CD16+ natural killer cells,a similar phenotype of EBV-CTL cultures from PBMCs.Interestingly,TIL cultures secreted high levels of the Th1 cytokines,interferon gamma (IFNγ) and tumor necrosis factor-alpha (TNF-α),and low levels of the Th2 cytokines,IL-4 and IL10.Moreover,young TILs could recognize autologous EBV-transformed B lymphoblast cell lines,but not autologous EBV-negative blast cells or allogeneic EBV-negative tumor cells.Taken together,these data suggest that ex vivo expansion of TILs from NPC biopsy tissue is an appealing alternative method to establish T cell-based immunotherapy for NPC.

  16. In vitro cytotoxic, genotoxic and antioxidant/oxidant effects of guaiazulene on human lymphocytes

    Directory of Open Access Journals (Sweden)

    Başak Toğar

    2015-02-01

    Full Text Available The aim of this study was to evaluate for the cytotoxicity, genotoxicity and antioxidant/oxidant activity of GYZ on human peripheral blood lymphocytes (PBLs. Guaiazulene (GYZ was added into culture tubes at various concentrations (0-400 µg/mL-1. Cytotoxicity against the human lymphocytes cultures was examined by lactate dehydrogenase (LDH release assay. The proliferative response was estimated by 3-(4,5-dimethylthiazol-2-yl-2,5-diphenyl tetrazolium bromide (MTT assay. Antioxidant/oxidant activity was evaluated by measuring the total oxidant status (TOS and total antioxidant capacity (TAC levels. Micronucleus (MN and chromosomal aberration (CA tests were used in genotoxicity studies. The results showed that GYZ caused cytotoxicity in the PBLs at high concentrations, but TOS level were not affected, while the level of TAC was significantly increased. GYZ also did not induce chromosomal aberrations when compared to that of the control group. Results this study clearly revealed that GYZ was not genotoxic and also increased the capacity of the antioxidant in the culture of human PBL cells. This report is first report on the impact of GYZ on human PBL cells.

  17. Migration of lymphocytes on fibronectin-coated surfaces: temporal evolution of migratory parameters

    Science.gov (United States)

    Bergman, A. J.; Zygourakis, K.; McIntire, L. V. (Principal Investigator)

    1999-01-01

    Lymphocytes typically interact with implanted biomaterials through adsorbed exogenous proteins. To provide a more complete characterization of these interactions, analysis of lymphocyte migration on adsorbed extracellular matrix proteins must accompany the commonly performed adhesion studies. We report here a comparison of the migratory and adhesion behavior of Jurkat cells (a T lymphoblastoid cell line) on tissue culture treated and untreated polystyrene surfaces coated with various concentrations of fibronectin. The average speed of cell locomotion showed a biphasic response to substrate adhesiveness for cells migrating on untreated polystyrene and a monotonic decrease for cells migrating on tissue culture-treated polystyrene. A modified approach to the persistent random walk model was implemented to determine the time dependence of cell migration parameters. The random motility coefficient showed significant increases with time when cells migrated on tissue culture-treat