Murine
To explain the life-long persistence of certain viruses in infected animals, Burnet and Fenner 1 ; proposed the hypothesis of immunologic tolerance. They suggested that after a generalized nonfatal infection of the embryo, the animal, after birth, would be incapable of producing antibody to the same infecting agent. Observations with lymphocytic choriomeningitis LCM; refs. 2-6 ; and tumor viruses 7-10 ; provided the main support for this concept. In both of these infections, virus is transmitted vertically and a permanent viral carrier state ensues. Characteristically, virus is detectable in blood and organs throughout life, while free circulating antibody to viral antigen s ; is not. Previously we reported that mice infected with LCM virus either in utero or in the newborn period were not "immunologically tolerant", as they were capable of making an immune response to the virus 11-13 ; . Now we report that neither immunologic nor clinical tolerance to Gross murine leukemia virus- GMuLV- ; related antigens occurs in AKR mice. Such mice are naturally infected in utero with GMuLV and carry large amounts of virus throughout their life 14-16.
Endogenous murine leukemia viruses MuLV's ; can be divided into two classes with respect to host range 1 ; . The ecotropic MuLV infects and replicates in mouse cells but not in heterologous cells. Conversely, the xenotropic MuLV's MuX's ; are not infectious for most mouse cell lines but can replicate well in cells from other species 18 ; . Integrated proviral DNAs of xenotropic viruses are present in the genomes of inbred mouse strains and wild mice, whereas only some strains have complete ecotropic MuLV proviral DNA 1, 6, 15 ; . Although the molecular structures of these endogenous proviruses are not well characterized, recent studies have revealed the basic organization of the unintegrated proviruses and integrated proviral DNA acquired by infection 8, 22, 28, ; . These studies suggest that MuLV proviral DNA is remarkably similar to the structures of bacterial and eucaryotic transposable elements 29 ; . The endogenous xenotropic viruses are of particular interest because of their putative parental role in the formation of recombinant viruses, which include Friend spleen focus-forming virus, the mink cell focus-inducing MCF ; viruses 9, 11, 24, ; , and the B-tropic MuLV's 2, 14 ; . All of these viruses have an altered host range and appear to have been formed by the recombination of ecotropic and xenotropic MuLV genomes 2, 9, 30 ; . The MCF-type viruses, which include recombinants of Moloney MuLV 10 ; , appear to be recombinants in the env gene region of the.
Since this paper was submitted for publication, another study of 5-HT synthesis by murine neuroblastoma cells in vitro has been reported by S. Knapp and A. J. Mandell 1974. Brain Res. 66: 574. ; . In addition, P. Mandel et al. 1973. In Frontiers in Catecholamine Research. E. Usdin and S. Snyder, editors. Pergamon Press Inc., Elmsford, N. Y. 277. ; , reported finding NE, E, DA, normetanephrine, histamine, and 5-HT by thinlayer chromatography of dansyl derivatives extracted from a clone of murine neuroblastoma cells. 69.
Graphite Low Profile Casting Reel Seat - Centered Bore: Trigger is a molded extension of the body, providing a more comfortable, secure grip. These tough graphite nylon reel seats are up to 50% lighter than all metal seats and also provide more corrosion resistance. Self-aligning stainless steel hood with a keyway on body, allows faster and stronger reel mounting. Ribbed interior bore to assure proper bonding with blank. Hood comes with a graphite nylon cushion which insulates reel feet, preventing electrolytic corrosion.
Mouse vas deferens protein MVDP ; is an aldose reductase-like protein that is highly expressed in the vas deferens and adrenal glands and whose physiological functions were unknown. We hereby describe the enzymatic characteristics of MVDP and its role in murine adrenocortical Y1 cells. The murine aldose reductase AR ; and MVDP cDNAs were expressed in bacteria to obtain recombinant proteins and to compare their enzymatic activities. Recombinant MVDP was functional and displayed kinetic properties distinct from those of murine AR toward various substrates, a preference for NADH, and insensitivity to AR inhibitors. For MVDP, isocaproaldehyde, a product of side-chain cleavage of cholesterol generated during steroidogenesis, is the best natural substrate identified so far. In Y1 cells, we found that NADH-linked isocaproaldehyde reductase ICR ; activity was much higher than NADPH-linked ICR activity and was not abolished by AR inhibitors. We demonstrate that in Y1 cells, forskolin-induced MVDP expression enhanced NADH-linked ICR activity by 5 6-fold, whereas no variation in ICR-linked NADPH activity was observed in the same experiment. In cells stably transfected with MVDP antisense cDNA, NADH-linked ICR activity was abolished even in the presence of forskolin, and the isocaproaldehyde toxicity was increased compared with that of intact Y1 cells, as measured by isocaproaldehyde LD50. In Y1 cells transfected with MVDP antisense cDNA, forskolin-induced toxicity was abolished by aminoglutethimide. These results indicate that in adrenocortical cells, MVDP is responsible for detoxifying isocaproaldehyde generated by steroidogenesis.
Heck KE; Schoendorf KC; Chavez GF; Braveman P; Does postpartum length of stay affect breastfeeding duration? A population-based study. 2003 30: 3 ; Birth and muse.
Institutes of health has been one of tag's key goals.
Rituximab is a chimeric anti-CD20 IgG1 kappa monoclonal antibody that contains murine variable regions and human constant regions.11 Although FDA-approved for the treatment of low-grade lymphoma, rituximab has single-agent activity in DLBCL, with an overall response rate of approximately 30-35% in pretreated patients.12, 13 Recent studies suggest that adding rituximab to CHOP significantly improves the CR rate and survival in patients with untreated DLBCL.14, 15 and mycostatin.
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Materials. Doxorubicin was from Sigma-Aldrich St. Louis, MO epirubicin Ellence; Pharmacia and Upjohn, Peapack, NJ ; was from the University of North Carolina at Chapel Hill Memorial Hospital Pharmacy; and pegylated liposomal doxorubicin Doxil ; was from Ortho Biotech Products, L.P. Bridgewater, NJ ; . Phosphatase inhibitors deltamethrin, nodularin, and okadaic acid were from Calbiochem San Diego, CA ; , as was the MEK inhibitor 2- 2-amino-3methoxyphenyl ; -4H-1-benzopyran-4-one PD 98059 ; , while sodium orthovanadate was from Sigma-Aldrich. The protease cocktail Complete was from Roche Applied Science Indianapolis, IN ; , and phenylmethylsulfonyl fluoride was from Fisher Scientific Co. Fair Lawn, NJ ; . Stock solutions were prepared in isopropanol for phenylmethylsulfonyl fluoride, Dulbecco's PBS from the Lineberger Comprehensive Cancer Center Tissue Culture Facility LCCC TCF, Chapel Hill, NC for sodium orthovanadate, or dimethyl sulfoxide for all others, and stored at 20C. These reagents were used at concentrations indicated in the text, with a final vehicle concentration that did not exceed 0.5% v v ; . All other chemicals, unless otherwise indicated, were obtained from Fisher Scientific Co. Cell Lines and Cell Culture. A1N4-myc cells were grown in Richter's modified Eagle's medium MEM ; with insulin, gentamicin, and 20 mM Hepes LCCC TCF ; , and further supplemented with 10 ng ml human recombinant EGF, 9% fetal bovine serum, 100 units ml penicillin G sodium, 100 g ml streptomycin sulfate these reagents from Invitrogen, Carlsbad, CA ; , and hydrocortisone Sigma-Aldrich ; . MDA-MB-231 and BT-474 human breast carcinoma cells LCCC TCF ; were grown in Richter's MEM as above except without EGF and hydrocortisone. Mouse embryo fibroblasts from heterozygous and homozygous MKP-1 knockout mice Dorfman et al., 1996 ; , as well as wild-type controls, generously provided by Dr. Naomi Laing and the Bristol Myers Squibb Research Institute Princeton, NJ ; , were cultured in Dulbecco's MEM. All cells were propagated in incubators providing a humidified atmosphere with 5% CO2 in air. Western Blotting. Adherent cells were detached by exposure to trypsin Invitrogen ; , counted using a hemacytometer Hausser Scientific; Horsham, PA ; , and 2 106 cells were seeded onto 60 mm Falcon 3002 tissue culture plates BD Labware; Lincoln Park, NJ ; in complete medium. These cells were allowed to reattach overnight and subjected to conditions described in the text, along with the addition of fresh medium. At the completion of an experiment all plates were placed on ice, washed with 5 ml of ice-cold PBS, and collected by scraping into eukaryotic lysis buffer as described previously Orlowski et al., 2002 ; . Relative protein concentrations of each sample were determined using the BCA protein assay kit Pierce Chemical, Rockford, IL ; . Equivalent protein amounts were subjected to Western blotting and immunoreactive bands were detected as described Orlowski et al., 2002 ; . For repeated analyses of the same filter, blots were stripped for 45 min using Western Re-Probe Geno Technology, Inc., St. Louis, MO ; following the manufacturer's specifications. Activation status of the p44 42-MAPKs was determined using murine monoclonal antibodies recognizing active, dually phosphorylated p44 42 Thr202 Tyr204 ; , with rabbit p44 42 antibodies used as loading controls. The activation status of the mitogen-activated protein kinase kinase MEK ; was determined using rabbit polyclonal antibodies recognizing active, dually phosphorylated MEK-1 2 Ser217 Ser221 ; , with rabbit MEK-1 2 antibodies used as loading controls. p90 RSK phosphorylation status was evaluated using rabbit polyclonal antibodies recognizing active, dually phosphorylated Thr359 Ser363 ; RSK, and Bad phosphorylation status was evaluated using a murine monoclonal antibody recognizing phosphoSer112-Bad all from Cell Signaling Technology, Inc., Beverly, MA ; . Total RSK levels were determined by Western blotting with a murine monoclonal anti-RSK antibody BD Biosciences, San Jose, CA ; , and Bad levels were evaluated with a rabbit polyclonal anti-Bad antibody Cell Signaling Technology, Inc. ; . Expression levels of the.
WU H, JERSE AE Department of Microbiology and Immunology, Uniformed Services University of Health Sciences, Bethesda, Maryland 20814-4799, USA Introduction: Gonococci in urethral exudates or blood demonstrate increased resistance to killing by normal human serum NHS ; , which is lost by treatment with neuraminidase NANase ; or upon subculture. Serum resistance can be restored by incubation with cytidine monophosphate neuraminic acid CMP-NANA ; . This unstable type of serum resistance results from the addition of sialic acid to the terminal N-lactose-N-tetraose molecule present on some gonococcal LOS species. Host sialic acid is used as the substrate in vivo, and the reaction is catalyzed by gonococcal sialyltransferase. In addition to conferring increased serum resistance, LOS sialylation increases resistance to opsonophagocytosis and to bactericidal activity of porin-specific antibodies. Collectively, these observations suggest that LOS sialylation confers a survival advantage. Sialylation also reduces gonococcal invasion of epithelial cells, however, a finding that suggests a more complex role for LOS sialylation in infection. Here we report the construction of a genetically defined sialyltransferase mutant, and evidence that LOS sialylation occurs during experimental murine infection in an effort to define the potential of the murine model for studying this aspect of gonococcal pathogenesis. Methods and Results: The sialyltransferase gene of N. gonorrhoeae strain MS11 was cloned and mutated via the insertion of a nonpolar kanamycin resistance cassette. A sialyltransferase-deficient mutant GP300 ; was constructed by allelic exchange. As shown by others, wild type MS11 was more resistant to NHS after culturing in the presence of CMP-NANA, and serum resistance was reduced after incubation with NANase bactericidal50 titers: wild type, 3%; wild type + CMP-NANA, 16%; wild type + CMP-NANA + NANase, 4% ; . In contrast, mutant GP300 was equally sensitive to NHS regardless of growth conditions or NANase treatment bactericidal50 titers: 3-4% for all conditions ; . To determine if LOS sialylation occurs during experimental murine infection, estradiol-treated BALB c mice were inoculated intravaginally with wild type MS11 or mutant GP300, and bactericidal assays were performed directly on gonococci in vaginal swab suspensions collected at selected time points. Gonococci from mice infected with wild type MS11 demonstrated increased resistance to normal human serum. Incubation of vaginal swab suspensions with NANase resulted in a serum sensitive phenotype bactericidal50 titers: 16% and 2-4%, respectively ; . Serum resistance was detected as early as 2 days following intravaginal inoculation with wild type MS11, and was lost upon subculture to artificial media. In contrast, gonococci in vaginal swab suspensions from mice infected with mutant GP300 were sensitive to NHS bactericidal50 titer: 2-4% ; , regardless of the time point at which they were collected. Conclusions: These results are indirect evidence that sialylation of gonococcal LOS occurs in the lower genital tract of mice as early as two days after inoculation. The estradiol-treated mouse model may therefore be useful as a tool for studying LOS sialylation in vivo and mysoline.
Plasma membrane of mature sialylated Ii forms, would imply that newly synthesized Ii complexes reside for a very short time in endosomes before cell surface appearance in DCs. Interestingly, we demonstrate that within 30 min after the internalization, most of the cell surface Ii complexes 74% ; are al.
19. LeBlanc R, Catley LP, Hideshima T, et al. Proteasome inhibitor PS-341 inhibits human myeloma cell growth in vivo and prolongs survival in a murine model. Cancer Res 2002; 62: 4996 Kaluz S, Kaluzova M, Stanbridge EJ. Proteasomal inhibition attenuates transcriptional activity of hypoxia-inducible factor 1 HIF-1 ; via specific effect on the HIF-1a C-terminal activation domain. Mol Cell Biol 2006; 26: 5895 Fidler IJ. Regulation of neoplastic angiogenesis. J Natl Cancer Inst Monogr 2001; 28: 10 Kassouf W, Dinney CP, Brown G, et al. Uncoupling between epidermal growth factor receptor and downstream signals defines and nadolol.
This Cer tif i cate de scribes ben e fits un der the Catamount Blue health plan. Catamount Blue is a Preferred Provider Organization PPO ; plan issued by Blue Cross and Blue Shield of Vermont. Catamount Blue provides better benefits when you use health care Providers in the Catamount Blue Preferred Provider Network. If you use Providers who are not in the Preferred Network Non-preferred Providers ; , you pay a higher share of the cost of your care. Chapter One explains what you must do to get benefits through your health plan. Your Outline of Coverage shows what you must pay. Read this entire chapter carefully, as it is your responsibility to follow the guidelines it lists.
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Table 2. Gas-Chromatographlc Analysis for Biliary Lipids and nafcillin.
Monoclonal antibodies and scFv antibody fragment MAb TH69 is a murine IgG1 with specificity for human CD73 and was kindly provided by Prof. Dr. Martin Gramatzki, Division of Stem Cell and Immunotherapy, 2nd.
Production by immature DC 14. To address the potential role that the above cytokines may play in the action of IVIg, we analysed the treatment of thrombocytopenia in mice lacking key pro-inflammatory regulators that could potentially affect DC or macrophage function Fig 2A ; . Mice lacking TNF- , IL-12 , MIP1 , or IFN- R were rendered thrombocytopenic as in Fig. 1B, and all mice fully responded to IVIg treatment despite the lack of individual expression of these cytokines. The anti-inflammatory cytokine IL-10 can play an important role in dampening macrophage activation 23, and its production by DC is upregulated in response to IVIg in vitro 14. The anti-inflammatory cytokine IL-4 can induce the production of IL-1Ra 24, and upregulate expression of the inhibitory IgG receptor Fc RIIB 25, which is absolutely required for IVIg to ameliorate murine models of ITP 26, 27. We found, however, that IVIg did function therapeutically in IL-10 deficient mice as well as in IL-4 deficient mice Fig 2B and naloxone.
Collagens in osteochondrosis Studies on articular cartilage have revealed the possible functions of some of the minor collagens. Type IX is found attached to the surface of type II fibrils Eyre et al., 1987; Vaughan et al., 1988 ; and strong evidence exists that it has a role in the regulation of fibril diameter Wotton et al., 1988 ; . Type VI collagen may be involved in maintaining the structural integrity of the tissue Bonaldo et al., 1990 ; and is seen to be elevated in diseases such as osteoarthritis Ronziere et al., 1990 ; . Type X collagen is found exclusively in the hypertrophic region of growth plate cartilage Gibson and Flint, 1985 ; and is only rarely detected in the calcifying region of articular cartilage Gannon et al., 1988 ; . It is found associated with type II collagen Chen et al., 1990 ; and its location suggests an important role in the process of calcification or chondrocyte maturation. Collagen undergoes numerous post-translational modifications, one of which is the formation of intermolecular crosslinks, which confers strength and stability to connective tissues Eyre et al., 1984 ; . A disturbance in the normal processes of cross-linking could result in a structurally weakened tissue as can be seen in osteochondritis dissecans. Conversely, an excessively cross-linked cartilagenous matrix could be resistant to the processes of resorption that occur at the ossification front giving rise to a lack of endochondral ossification. Osteochondrosis is not only a disease of considerable economic importance but it also may be useful as a model with which to investigate endochondral ossification. The purpose of this study is to establish whether changes in the nature, ratio or distribution of the collagenous components of the articular and growth plate cartilages occur during osteochondrosis. MATERIALS AND METHODS.
COLONIAL ERA Introduction of Settlement at Williams Lake The rapid settlement of the Williams Lake area followed the local discovery of gold in the late 1850s and what came to be known as the Fraser Gold Rush. Settlements were established in the 1860s as the prospectors turned to ranching and logging.46 This development represented a significant change for the First Nations in the area, whose only other previous contact with nonFirst Nation people had been related to the fur trade and with Catholic and Oblate missions.47 According to historical documentation, the first recorded encounter between the Williams Lake Indian Band and the white man took place in January 1842, when the Oblate missionary Modeste Demers visited the First Nation in its village at Missioner Creek apparently on his second visit to the area. Reporting on this visit, Demers stated that the First Nation had been building houses and naltrexone.
FIGURE 3. Effects of IL-13 or IL-4 plus IL-13 deficiency on pulmonary fibrosis. The whole lung homogenates were prepared from BLM- or salinetreated mice at day 21 after treatment. They were then analyzed for hydroxyproline content and the results expressed as micrograms per lung. Relative to that in wild-type wt ; mice, the BLM-induced increase in lung hydroxyproline content was significantly reduced p 0.05 ; in both IL13-deficient IL-13 ko ; and IL-4 IL-13 doubly deficient IL-4 IL-13 ko ; mice. The levels of lung hydroxyproline in both BLM-treated knockout murine strains were not significantly different from their respective salinetreated controls. For each murine strain, mean SE of five animals per treatment group are shown.
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Tse KF, Mukherjee G, Small D. Constitutive activation of FLT3 stimulates multiple intracellular signal transducers and results in transformation. Leukemia. 2000; 14: 17661776. ; Fenski R, Flesch K, Serve S et al. Constitutive activation of FLT3 in acute myeloid leukaemia and its consequences for growth of 32D cells. Br J Haematol. 2000; 108: 322330. ; Kelly LM, Liu Q, Kutok JL et al. FLT3 internal tandem duplication mutations associated with human acute myeloid leukemias induce myeloproliferative disease in a murine bone marrow transplant model. Blood. 2002; 99: 310-318. ; Kelly LM, Kutok JL, Williams IR et al. PML RARalpha and FLT3-ITD induce an APLlike disease in a mouse model. Proc Natl Acad Sci U S A. 2002; 99: 8283-8288. ; Druker BJ, Talpaz M, Resta DJ et al. Efficacy and safety of a specific inhibitor of the bcrabl tyrosine kinase in chronic myeloid leukemia. N Engl J Med. 2001; 344: 1031-1037. ; Druker BJ, Sawyers CL, Kantarjian H et al. Activity of a specific inhibitor of the BCRABL tyrosine kinase in the blast crisis of chronic myeloid leukemia and acute lymphoblastic leukemia with the Philadelphia chromosome. N Engl J Med. 2001; 344: 1038-1042. ; Kelly LM, Yu JC, Boulton CL et al. CT53518, a novel selective FLT3 antagonist for the treatment of acute myelogenous leukemia AML ; . Cancer Cell. 2002; 1: 421-432. ; Levis M, Tse KF, Smith BD, Garrett E, Small D. A FLT3 tyrosine kinase inhibitor is selectively cytotoxic to acute myeloid leukemia blasts harboring FLT3 internal tandem duplication mutations. Blood. 2001; 98: 885-887. ; Weisberg E, Boulton C, Kelly LM et al. Inhibition of mutant FLT3 receptors in leukemia cells by the small molecule tyrosine kinase inhibitor PKC412. Cancer Cell. 2002; 1: 433443. ; Yee KW, O'Farrell AM, Smolich BD et al. SU5416 and SU5614 inhibit kinase activity of wild-type and mutant FLT3 receptor tyrosine kinase. Blood. 2002; 100: 2941-2949. ; O'Farrell AM, Abrams TJ, Yuen HA et al. SU11248 is a novel FLT3 tyrosine kinase inhibitor with potent activity in vitro and in vivo. Blood. 2003; 101: 3597-3605. ; Zheng R, Friedman AD, Small D. Targeted inhibition of FLT3 overcomes the block to myeloid differentiation in 32Dcl3 cells caused by expression of FLT3 ITD mutations. Blood. 2002; 100: 4154-4161. ; Levis M, Allebach J, Tse KF et al. A FLT3-targeted tyrosine kinase inhibitor is cytotoxic to leukemia cells in vitro and in vivo. Blood. 2002; 99: 3885-3891. ; Stone RM, De Angelo J, Galinsky I et al. PKC 412 FLT3 inhibitor therapy in AML: results of a phase II trial. Ann Hematol. 2004; 83 Suppl 1: S89-S90. 21 ; Heinrich MC, Druker BJ, Curtin PC et al. A "First in Man" Study of the Safety and PK PD of Oral FLT3 Inhibitor MLN518 ; in Patients with AML or High Risk Myelodysplasia [abstract]. Blood. 2002; 100: 336a-337a. ; Smith BD, Levis M, Beran M et al. Single agent CEP-701, a novel FLT3 inhibitor, shows biologic and clinical activity in patients with relapsed or refractory acute myeloid leukemia. Blood. 2004. 23 ; Lowenberg B, Downing JR, Burnett A. Acute myeloid leukemia. N Engl J Med. 1999; 341: 1051-1062. ; Yokota S, Kiyoi H, Nakao M et al. Internal tandem duplication of the FLT3 gene is preferentially seen in acute myeloid leukemia and myelodysplastic syndrome among and namenda.
Tracheally. One or two hundred micrograms per animal of sheep anti-mouse IL-12 monoclonal antibody, provided by Genetics Institute Cambridge, MA ; , or 100 g animal of nonspecific sheep IgG ICN Pharmaceuticals, Aurora, OH ; were injected intravenously immediately after bleomycin instillation and 7 days after instillation. The anti-IL-12 antibody used in this research was generated by immunizing sheep with intact p70 murine IL-12 and binds to the p40 subunit of IL-12 as well as to the intact p70 heterodimer but not to the p35 subunit according to the manufacturer's instructions. After treatment, the mice were returned to their cages and allowed food and water ad libitum. The mice were anesthetized either 7 or 14 days after bleomycin instillation and then killed. Samples of the right lung were excised to be processed for light microscopy. Samples of the left lung tissue were frozen in liquid nitrogen for the measurement of hydroxyproline content. Cell differential counts in bronchoalveolar lavage fluid. A tracheotomy was performed in killed mice. After insertion of a tracheal tube, the trachea was lavaged five times with 1 ml of sterile saline at room temperature. The recovered fluid was filtered through a single layer of gauze to remove mucus. The cells in the bronchoalveolar lavage BAL ; fluid BALF ; were.
Hamster buccal pouch carcinoma has proved to be a valuable experimental model for the study of different parameters of oral mucous membrane malignancy Salley, 1954; Shklar, 1968 ; . Major advantages of this model include: the predictable induction of tumors by topical application of dimethylbenzanthracene DMBA the development of pre-cancerous dysplastic lesions comparable with human oral leukoplakia erythroplakia; and the histological similarity of the induced carcinoma to the well-differentiated type of human squamous cell carcinoma of the oral cavity. One of the disadvantages of this system is that the tumors are of the exophytic type and have a tendency to proliferate on the surface rather than invade the underlying tissues and metastasize as rapidly as do many human oral carcinomas. Although cervical lymph node metastases of the hamster buccal pouch carcinomas were reported in the first study by Salley 1954 ; , later investigators Shklar, 1966; Levij et al., 1967; Giunta and Shklar, 1971; Sheehan et al., 1971; Eveson, 1981 ; were unable to find metastases in their studies. One case of lymph node metastasis was published by Rwomushana et al. 1970 ; as an extremely rare finding. So it has generally been thought that chemically induced squamous cell carcinomas characteristically do not metastasize to the lymph nodes. However, two recent studies Craig, 1980; Safour et al., 1984 ; demonstrated that, under special experimental conReceived for publication July 17, 1985 Accepted for publication February 20, 1986 * Based on a thesis submitted to the graduate faculty, Loyola University School of Dentistry, in partial fulfillment of the requirements for the M.S. in Oral Biology degree and naratriptan and murine.
31. McCormack, J. E., T. Wade, H. Morales, J. Kappler, and P. Marrack. 1991. Analysis of class II MHC structure in thymic nurse cells. Cell. Immunol. 138: 413422. 32. Nakashima, M., K. Mori, K. Maeda, H. Kishi, K. Hirata, M. Kawabuchi, and T. Watanabe. 1990. Selective elimination of double-positive immature thymocytes by a thymic epithelial cell line. Eur. J. Immunol. 20: 4753. 33. Nishimura, T., Y. Takeuchi, Y. Ichimura, X. Goa, A. Akatsuka, N. Tamaoki, H. Yagita, K. Okumura, and S. Habu. 1990. Thymic stromal cell clone with nursing activity supports the growth and differentiation of murine CD4 8 thymocytes in vitro. J. Immunol. 145: 40124017. 34. Ohashi, P. S., H. Pircher, K. Burki, R. M. Zinkernagel, and H. Hengartner. 1990. Distinct sequence of negative or positive selection implied by thymocyte T-cell receptor densities. Nature 346: 861863. 35. Penit, C. 1986. In vitro thymocyte maturation. Budr labeling of cycling thymocytes and phenotypic analysis of their progeny support the single lineage model. J. Immunol. 137: 21152121. 36. Penninger, J., T. Rieker, N. Roman, J. Klima, W. Salvenmoser, H. Dietrich, H. Stossel, and G. Wick. 1994. Ultrastructural analysis of thymic nurse cell epithelium. Eur. J. Immunol. 24: 222228. 37. Penninger, J., and G. Wick. 1992. Thymic nurse cell lymphocytes react against self-major histocompatibility complex. Eur. J. Immunol. 22: 7983. 38. Pezzano, M., Y. Li, Y. Yang, and J. Guyden. 1991. The immortalization of thymic nurse cells by SV40 virus. Cell. Immunol. 133: 434445. 39. Pezzano, M., D. Philp, S. Stephenson, Y. Li, V. Reid, R. Maitta, and J. C. Guyden. 1996. Positive selection by thymic nurse cells requires IL-1 beta and is associated with an increased Bcl-2 expression. Cell. Immunol. 169: 174 184. Pezzano, M., Y. Li, D. Philp, C. Omene, M. Cantey, G. Saunders, and J. C. Guyden. 1995. Thymic nurse cell rescue of early CD4 CD8 thymocytes from apoptosis. Cell Mol. Biol. 41: 10991111. 41. Pezzano, M., K. D. King, D. D. Philp, A. Adeyemi, B. Gardiner, J. Yang, M. Samms, W. Boto, and J. C. Guyden. 1998. A thymic nurse cell-specific monoclonal antibody. Cell. Immunol. 185: 123133. 42. Philp, D., M. Pezzano, Y. Li, C. Omene, W. Boto, and J. C. Guyden. 1993. The binding, internalization, and release of thymocytes by thymic nurse cells. Cell. Immunol. 148: 301315. 43. Rieker, T., J. Penninger, N. Roman, and G. Wick. 1995. Chicken thymic nurse cells: an overview. Dev. Comp. Immunol. 19: 281289. 44. Ritter, A., C. A. Suavage, and S. F. Cotmore. 1981. The human thymus microenvironment: in vivo identification of thymic nurse cells and other antigenically-distinct subpopulations of epithelial cells. Immunology 44: 439 446. Roberts, F., V. Geenen, J. Schoenen, E. Burgeon, P. Defresne, J. J. Legros, and P. Franchimont. 1991. Colocalization of immunoreactive oxytocin, va.
The lamina propria. In general, the severity of inflammation detected by immunohistological analysis correlated with the clinical score in SB203580-treated and nontreated animals. No lesions or signs of inflammation were observed in control mice. SB203580 targets only the ATP binding of p38 but does not interfere with p38 phosphorylation 44 ; . To correctly assess the activity of p38, we immunoprecipitated this MAPK from bowel tissue protein extracts and performed an in vitro kinase assay using ATF-2 as a substrate followed by immunoblotting with phospho-specific ATF-2 antibodies. A strong increase of p38 activity was detected after day 3 with a maximum between days 5 and 10. This activity was strongly inhibited by SB203580 during the course of colonic inflammation Fig. 4B ; . SB203580 inhibits the classical and alternative NF-B pathways In addition to p38, NF-B is an important regulator of genes encoding cytokines and their receptors. SB203580 inhibited these cytokine genes in murine bowel tissue. Therefore, we wanted to examine whether SB203580 had also an effect on NF-B activation in our animal model of intestinal inflammation. Cytoplasmic and nuclear extracts were prepared from bowel tissue. First, we analyzed the phosphorylation of IKK, IB degradation, and the nuclear translocation of p65 by immunoblot analysis. These parameters served as a readout for NF-B activation. We found a statistically significant peak of IKK -phosphorylation and of IBdegradation at day 7 after the start of DSS feeding Fig. 5A ; . In addition, nuclear translocation of p65 showed the same kinetics as IB degradation Fig. 5B ; . Both observations indicate a maximal NF-B activation at days 5 to 7. Surprisingly, SB203580 inhibited IB degradation and p65 translocation, respectively, indicating that SB203580 is not only specific for p38 MAPK but has also other targets involved in NF-B activation. We further analyzed the alternative pathway of NF-B activation involving p100 processing to active p52. We performed immunoblot analysis using antibodies recognizing both, the p100 precursor as well as the p52 product. This NF-B activation pathway was also activated during DSS-induced colitis. Furthermore, the p100 processing was strongly blocked by SB203580 treatment Fig. 5C ; . This indicates that SB203580 has an inhibitory effect on both NF-B activation pathways. To analyze putative targets of SB203580, which exert a role in NF-B activation, we studied molecular components regulated by NOD proteins. Activation of NOD, intracellular receptor proteins for bacterial peptidoglycan, plays a crucial role in the pathogenesis of IBD 26 ; by triggering intestinal inflammation 36 ; . RICK is a critical downstream kinase of NOD2 and regulates IKK finally leading to NF-B activation 45 ; . Therefore, we assessed RICK activity during our experimental colitis induction. Immunoprecipitated RICK from protein extracts derived from bowel tissue phosphorylates MBP in an in vitro kinase assay Fig. 5D ; . Those assays indicated that RICK is dramatically activated during development of experimentally induced ulcerative colitis. Activation of RICK can be almost completely inhibited by SB203580 treatment of the mice Fig. 5D ; . Thus, on the basis of our data, the inhibition of RICK by SB203580 may be responsible for the efficient suppression of NF-B activation and the release of proinflammatory cytokines and narcan.
CD8 T cells in a pulmonary CD62Llo memory-effector cell pool during latent murine cytomegalovirus infection of the lungs. Journal of Virology 74, 1149511503.
In Project II, the effect of dose and application frequency on the in vivo penetration of TACA into stratum corneum was investigated in 15 healthy volunteers. Dose and application frequency of topical corticosteroids are recurrently debated topics. Multiple-daily applications are common, although a superior efficacy compared to once-daily applications is not unequivocally proven. In the dose experiment, higher TACA amounts were quantified within the stratum corneum after application of a high dose 300 g cm vs. 100 g cm ; acetonic solution ; . However, this.
And found that conjugating capsule to bovine serum albumin and PA increased its antigenicity. Determined in preliminary experiments that B. anthracis capsule may have minor adjuvant properties. Initiated experiments with blocking monoclonal antibodies to known receptors on macrophages to determine if masking a known receptor inhibits spore binding and confirmed that the antibodies were truly bound to murine peritoneal macrophages. Constructed mutants in the putative adherence genes, adcA and orf168 of the B. anthracis Ames strain. Characterized a B. anthracis mutant in a rhamnose biosynthesis gene, spsI. Analyzed spores made from this mutant strain by transmission electron microscopy and found that most spores of the spsI mutant lacked an exosporium. In contrast, the exosporium was present on all spores from the Ames wild-type strain. Analyzed the carbohydrates of both the spsI mutant and Ames wild-type strains and found rhamnose in the exosporium from the Ames strain but not in the exosporium of the spsI mutant. Examined the interaction between B. anthracis spores and macrophages and recovered fewer colony-forming units CFU ; from the mutant strains than from the wild-type strain. Challenged 10 guinea pigs intramuscularly with 2000 spores of the mutant or the Ames wild-type strain but failed to observe any differences in survivability between the wildtype strain and mutant. Performed preliminary experiments to determine if anthrax spores are able to adhere A549 cells, a Type-II lung carcinoma line and found that spores adhered to the pneumocytes. Transmission electron microscopy suggested that a specific receptor exists for binding the spore to the pneumocyte. Defined this receptor by using lectincoated cells, which showed a dramatic increase in spore adherence.
Transfections HT29 cells were transfected with Lipofectamine Plus Invitrogen ; as previously described [3]. In transfections monitoring reporter gene expression, transfections were performed at least three times in 6 well dishes in triplicate with the total DNA mass kept constant at 4 g effector plasmid DNA, 100 ng 2 NF-B Luc reporter gene, 50 ng pRL-TK, a thymidine kinase promoter driven Renilla luciferase normalization reporter and 1.85 g pCDNA3.1 plasmid DNA as bulk filler DNA ; and fire-fly luciferase expression was normalized to Renilla luciferase expression using the dual-luciferase assay Promega, Madison, WI ; . Fold inductions were calculated and values between experiments did not vary more than 15%, a representative experiment is presented. Transfection of 293T cells was performed with lipofectamine 2000 Invitrogen ; in 6-well dishes in triplicate as per the manufacture's protocol. TLR expression plasmids were added at 2 g well, and NF-B and normalization control plasmids were as above with HT29 cells and pCDNA3.1 plasmid DNA as bulk filler DNA to a final DNA mass of 4 g well. Fold inductions were calculated and values between experiments N of 3 ; did not vary more than 10%, a representative experiment is presented. Real Reverse Transcription and Real Time PCR RT2PCR ; Cells N 3 ; were stimulated 3 hours at 37C with TNF or FliC or left untreated and harvested for total RNA isolation. Total cellular RNA was extracted from cells with Trizol reagent Invitrogen ; [3] and reverse transcribed with ReactionReady first strand cDNA synthesis kit SuperArray Bioscience Corp., Fredrick, MD ; . RNA 2.5 ug per 20 ul reaction ; was reverse transcribed using random primers and Moloney murine leukemia virus reverse transcriptase per manufacturer specified conditions. Controls without reverse transcriptase minus RT ; was also generated for each RNA sample. RT2PCR was performed with an iCycler Bio-Rad ; to quantify TLR1 through TLR10 mRNA, 18S rRNA, and GAPDH mRNA. RT2PCR 25 ul reaction volume ; was performed with the appropriate primers SuperArray ; per manufacturers instructions in triplicate with HotStart Taq DNA polymerase SuperArray ; at 95C for 15 min to activate Taq and amplified for 40 cycles 95C, 30 sec, 55C, 30 sec, 72C, 30 sec ; . RT2PCR was performed on the minus RT controls with TLR5 primers to detect DNA contamination. Real-time PCR analysis was performed using SYBR-green Perkin-Elmer ; according to manufacture's instructions with the specific primer pairs indicated above and primer pairs for 18S ribosomal RNA as reference RNA Classic 18S primer pairs Ambion Inc ; . Cycle time Ct ; was measured using the iCyclerTM and its associated software Bio-Rad ; . Relative transcript quantities were calculated by the Ct method using 18S ribosomal RNA as a reference amplified from samples using the Classic 18S primer pairs from Ambion, Inc Austin, TX.
Sch.336 attenuated monocyte macrophage, granulocyte and lymphocyte migration to a CCL2-soaked gelfoam sponge Fig.6 ; . This effect was dose dependent, unaffected by co-administration with a CB1-specific antagonist SR141716A and required that the triaryl bis-sulfone bind to the CB2 receptor Fig. 7 ; . These results, when coupled with the inability of Sch.336 to alter binding of CCL2 to CCR2 or to downregulate CCR2 expression data not shown ; , support our contention that Sch.336 exerts its bioactivity through the CB2 receptor. We next demonstrate that SCH 336 can dampen pleuritis induced in a murine model of delayed type hypersensitivity Fig. 8 ; . Finally, we show that oral administration of Sch.336 attenuates ovalbumin-induced lung eosinophilia, a system that mimics certain pathobiological hallmarks of human asthmatic disease. In each of these models, Sch.336 demonstrated efficacy similar to that seen with more traditional anti-inflammatory agents. These data underscore the potential utility of CB2selective inverse agonists in the treatment of human asthma Kasserra et al., 2004 ; as well as other inflammatory diseases and muse.
6. Propst-Proctor SL, DillinghamMF, McDougallIR, Goodwin D. The white blood cell scanin orthopedics.C in Orthop.
This study was designed to evaluate the possible complications following the use of intraoperative mitomycin-C in patients undergoing surgery for primary pterygium. Hundred eyes of 88 patients with primary pterygium were operated upon using ''bare sclera technique''. Intraoperatively all the patients were applied freshly prepared mitomycin-C 0.02% mg ml ; on the bare sclera for a period of 5 minutes, after which the eyes were thoroughly rinsed with a sterile balanced salt solution. Postoperatively, none of the patients recieved mitomycin-C eye drops. After a mean postoperative followup of 14 months, 4 eyes 4% ; of 4 patients developed thinning of the sclera with visualization of the underlying uveal tissue. One eye 1% ; developed extreme thinning of the sclera with ectasia of the underlying uveal tissue, requiring a scleral patch. Recurrence was seen in 2 eyes 2% ; only. The study shows that although intraoperative mitomycin-C 0.02% ; reduces the recurrence of pterygium, it leads to serious, cosmetically unacceptable and even sight-threatening complications.
Service-oriented computing [18] is emerging as a new promising computing paradigm that centres on the notion of service as the fundamental element for developing software applications. In this scenario, two prominent issues involved in the development of next generation distributed software applications can be roughly synthesised as: 1 ; discovering available services that can be exploited to build a needed application, and 2 ; suitably aggregating such services to achieve the desired result. A typical example [16] of the need of aggregating services is a client wishing to make all the arrangements necessary for a trip flights, hotel, rent-a-car, and so on ; . Such a client query may not be satisfied by a single service, while it could be satisfied by composing several services. Complex Web service interactions however require more than SOAP, WSDL and UDDI can offer [7], and semi-automatic aggregation frameworks based on such standards are not available yet. The aim of this paper is to contribute to the long-term objective of lifting service aggregation from manual hand-crafting to a semi-automated engineered process. We present a methodology which, given a set of service contracts, tries to construct an aggregation of such services. Service contracts include a description of the service behaviour expressed by a YAWL [23] workflow ; , as well as an ontology-annotated ; signature. The core aggregation process basically performs a control-flow and an ontology-aware ; data-flow analysis of a set of YAWL workflows to build the contract of an aggregated service. Technically, these analyses are defined by first expanding the services' workflows with dummy YAWL flow constructs, and by exploiting ontology-matching mechanisms to perform a semantics-aware data-flow analysis. It is worth noting that the aggregation.
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