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Sample records for flere unge til

  1. Structural and biophysical analysis of interactions between cod and human uracil-DNA N-glycosylase (UNG) and UNG inhibitor (Ugi)

    SciTech Connect

    Assefa, Netsanet Gizaw; Niiranen, Laila; Johnson, Kenneth A.; Leiros, Hanna-Kirsti Schrøder; Smalås, Arne Oskar; Willassen, Nils Peder; Moe, Elin

    2014-08-01

    A structural and biophysical study of the interactions between cod and human uracil-DNA N-glycosylase (UNG) and their inhibitor Ugi is presented. The stronger interaction between cod UNG and Ugi can be explained by a greater positive electrostatic surface potential. Uracil-DNA N-glycosylase from Atlantic cod (cUNG) shows cold-adapted features such as high catalytic efficiency, a low temperature optimum for activity and reduced thermal stability compared with its mesophilic homologue human UNG (hUNG). In order to understand the role of the enzyme–substrate interaction related to the cold-adapted properties, the structure of cUNG in complex with a bacteriophage encoded natural UNG inhibitor (Ugi) has been determined. The interaction has also been analyzed by isothermal titration calorimetry (ITC). The crystal structure of cUNG–Ugi was determined to a resolution of 1.9 Å with eight complexes in the asymmetric unit related through noncrystallographic symmetry. A comparison of the cUNG–Ugi complex with previously determined structures of UNG–Ugi shows that they are very similar, and confirmed the nucleotide-mimicking properties of Ugi. Biophysically, the interaction between cUNG and Ugi is very strong and shows a binding constant (K{sub b}) which is one order of magnitude larger than that for hUNG–Ugi. The binding of both cUNG and hUNG to Ugi was shown to be favoured by both enthalpic and entropic forces; however, the binding of cUNG to Ugi is mainly dominated by enthalpy, while the entropic term is dominant for hUNG. The observed differences in the binding properties may be explained by an overall greater positive electrostatic surface potential in the protein–Ugi interface of cUNG and the slightly more hydrophobic surface of hUNG.

  2. Rev1 is essential in generating G to C transversions downstream of the Ung2 pathway but not the Msh2+Ung2 hybrid pathway.

    PubMed

    Krijger, Peter Hugo Lodewijk; Tsaalbi-Shtylik, Anastasia; Wit, Niek; van den Berk, Paul Cornelius Maria; de Wind, Niels; Jacobs, Heinz

    2013-10-01

    Somatic hypermutation (SHM) and class switch recombination (CSR) of immunoglobulin (Ig) genes are initiated by the enzymatic deamination of cytosine (C) to uracil (U). Uracil-DNA-glycosylase (Ung2) converts uracils into apyrimidinic (AP) sites, which is essential for the generation of transversions (TVs) at G/C basepairs during SHM and for efficient DNA break formation during CSR. Besides Ung2, the mismatch repair protein Msh2 and the translesion synthesis (TLS) DNA polymerase (Pol) Rev1 are implicated in SHM and CSR. To further unravel the role of Rev1, we studied WT, Rev1-deficient, Msh2-deficient, and Rev1, Msh2 double-deficient B cells. Loss of Rev1 only slightly reduced CSR. During SHM G/C to C/G TVs are generated in both Ung2- and Ung+Msh2-dependent fashions. We found that Rev1 is essential for the Msh2-independent generation of these TVs downstream of Ung2-induced AP sites. In the Ung+Msh2 hybrid pathway, Rev1 is not essential and can be substituted by an alternative TLS Pol, especially when Rev1 is lacking.

  3. Opinion: uracil DNA glycosylase (UNG) plays distinct and non-canonical roles in somatic hypermutation and class switch recombination.

    PubMed

    Yousif, Ashraf S; Stanlie, Andre; Begum, Nasim A; Honjo, Tasuku

    2014-10-01

    Activation-induced cytidine deaminase (AID) is essential to class switch recombination (CSR) and somatic hypermutation (SHM). Uracil DNA glycosylase (UNG), a member of the base excision repair complex, is required for CSR. The role of UNG in CSR and SHM is extremely controversial. AID deficiency in mice abolishes both CSR and SHM, while UNG-deficient mice have drastically reduced CSR but augmented SHM raising a possibility of differential functions of UNG in CSR and SHM. Interestingly, UNG has been associated with a CSR-specific repair adapter protein Brd4, which interacts with acetyl histone 4, γH2AX and 53BP1 to promote non-homologous end joining during CSR. A non-canonical scaffold function of UNG, but not the catalytic activity, can be attributed to the recruitment of essential repair proteins associated with the error-free repair during SHM, and the end joining during CSR.

  4. The DDB1–DCAF1–Vpr–UNG2 crystal structure reveals how HIV-1 Vpr steers human UNG2 toward destruction

    PubMed Central

    Wu, Ying; Zhou, Xiaohong; Barnes, Christopher O.; DeLucia, Maria; Cohen, Aina E.; Gronenborn, Angela M.; Ahn, Jinwoo; Calero, Guillermo

    2017-01-01

    The HIV-1 accessory protein Vpr is required for efficient viral infection of macrophages and promotion of viral replication in T cells. Vpr’s biological activities are closely linked to the interaction with human DCAF1, a cellular substrate receptor of the Cullin4–RING E3 ubiquitin ligase (CRL4) of the host ubiquitin–proteasome-mediated protein degradation pathway. The molecular details of how Vpr usurps the protein degradation pathway have not been delineated. Here we present the crystal structure of the DDB1–DCAF1–HIV-1–Vpr–uracil-DNA glycosylase (UNG2) complex. The structure reveals how Vpr engages with DCAF1, creating a binding interface for UNG2 recruitment, in a manner distinct from the recruitment of SAMHD1 by Vpx protein for degradation by Vpx proteins. Vpr and Vpx use similar N-terminal and helical regions to bind the substrate receptor, whereas different regions target the specific cellular substrates. Furthermore, Vpr uses molecular mimicry of DNA by a variable loop for specific recruitment of the UNG2 substrate. PMID:27571178

  5. HIV-1 and HIV-2 exhibit divergent interactions with HLTF and UNG2 DNA repair proteins

    PubMed Central

    Hrecka, Kasia; Hao, Caili; Shun, Ming-Chieh; Kaur, Sarabpreet; Swanson, Selene K.; Florens, Laurence; Washburn, Michael P.; Skowronski, Jacek

    2016-01-01

    HIV replication in nondividing host cells occurs in the presence of high concentrations of noncanonical dUTP, apolipoprotein B mRNA-editing, enzyme-catalytic, polypeptide-like 3 (APOBEC3) cytidine deaminases, and SAMHD1 (a cell cycle-regulated dNTP triphosphohydrolase) dNTPase, which maintains low concentrations of canonical dNTPs in these cells. These conditions favor the introduction of marks of DNA damage into viral cDNA, and thereby prime it for processing by DNA repair enzymes. Accessory protein Vpr, found in all primate lentiviruses, and its HIV-2/simian immunodeficiency virus (SIV) SIVsm paralogue Vpx, hijack the CRL4DCAF1 E3 ubiquitin ligase to alleviate some of these conditions, but the extent of their interactions with DNA repair proteins has not been thoroughly characterized. Here, we identify HLTF, a postreplication DNA repair helicase, as a common target of HIV-1/SIVcpz Vpr proteins. We show that HIV-1 Vpr reprograms CRL4DCAF1 E3 to direct HLTF for proteasome-dependent degradation independent from previously reported Vpr interactions with base excision repair enzyme uracil DNA glycosylase (UNG2) and crossover junction endonuclease MUS81, which Vpr also directs for degradation via CRL4DCAF1 E3. Thus, separate functions of HIV-1 Vpr usurp CRL4DCAF1 E3 to remove key enzymes in three DNA repair pathways. In contrast, we find that HIV-2 Vpr is unable to efficiently program HLTF or UNG2 for degradation. Our findings reveal complex interactions between HIV-1 and the DNA repair machinery, suggesting that DNA repair plays important roles in the HIV-1 life cycle. The divergent interactions of HIV-1 and HIV-2 with DNA repair enzymes and SAMHD1 imply that these viruses use different strategies to guard their genomes and facilitate their replication in the host. PMID:27335459

  6. Proximity to AGCT sequences dictates MMR-independent versus MMR-dependent mechanisms for AID-induced mutation via UNG2.

    PubMed

    Thientosapol, Eddy Sanchai; Sharbeen, George; Lau, K K Edwin; Bosnjak, Daniel; Durack, Timothy; Stevanovski, Igor; Weninger, Wolfgang; Jolly, Christopher J

    2016-12-29

    AID deaminates C to U in either strand of Ig genes, exclusively producing C:G/G:C to T:A/A:T transition mutations if U is left unrepaired. Error-prone processing by UNG2 or mismatch repair diversifies mutation, predominantly at C:G or A:T base pairs, respectively. Here, we show that transversions at C:G base pairs occur by two distinct processing pathways that are dictated by sequence context. Within and near AGCT mutation hotspots, transversion mutation at C:G was driven by UNG2 without requirement for mismatch repair. Deaminations in AGCT were refractive both to processing by UNG2 and to high-fidelity base excision repair (BER) downstream of UNG2, regardless of mismatch repair activity. We propose that AGCT sequences resist faithful BER because they bind BER-inhibitory protein(s) and/or because hemi-deaminated AGCT motifs innately form a BER-resistant DNA structure. Distal to AGCT sequences, transversions at G were largely co-dependent on UNG2 and mismatch repair. We propose that AGCT-distal transversions are produced when apyrimidinic sites are exposed in mismatch excision patches, because completion of mismatch repair would require bypass of these sites.

  7. Cell cycle regulation as a mechanism for functional separation of the apparently redundant uracil DNA glycosylases TDG and UNG2

    PubMed Central

    Hardeland, Ulrike; Kunz, Christophe; Focke, Frauke; Szadkowski, Marta; Schär, Primo

    2007-01-01

    Human Thymine-DNA Glycosylase (TDG) is a member of the uracil DNA glycosylase (UDG) superfamily. It excises uracil, thymine and a number of chemical base lesions when mispaired with guanine in double-stranded DNA. These activities are not unique to TDG; at least three additional proteins with similar enzymatic properties are present in mammalian cells. The successful co-evolution of these enzymes implies the existence of non-redundant biological functions that must be coordinated. Here, we report cell cycle regulation as a mechanism for the functional separation of apparently redundant DNA glycosylases. We show that cells entering S-phase eliminate TDG through the ubiquitin–proteasome system and then maintain a TDG-free condition until G2. Incomplete degradation of ectopically expressed TDG impedes S-phase progression and cell proliferation. The mode of cell cycle regulation of TDG is strictly inverse to that of UNG2, which peaks in and throughout S-phase and then declines to undetectable levels until it appears again just before the next S-phase. Thus, TDG- and UNG2-dependent base excision repair alternates throughout the cell cycle, and the ubiquitin–proteasome pathway constitutes the underlying regulatory system. PMID:17526518

  8. Discovery of ATP-Competitive Inhibitors of tRNAIle Lysidine Synthetase (TilS) by High-Throughput Screening.

    PubMed

    Shapiro, Adam B; Plant, Helen; Walsh, Jarrod; Sylvester, Mark; Hu, Jun; Gao, Ning; Livchak, Stephania; Tentarelli, Sharon; Thresher, Jason

    2014-09-01

    A novel, ultrahigh-throughput, fluorescence anisotropy-based assay was developed and used to screen a 1.4-million-sample library for compounds that compete with adenosine triphosphate (ATP) for binding to Escherichia coli tRNA(Ile) lysidine synthetase (TilS), an essential, conserved, ATP-dependent, tRNA-modifying enzyme of bacterial pathogens. TilS modifies a cytidine base in the anticodon loop of Ile2 tRNA by attaching lysine, thereby altering codon recognition of the CAU anticodon from AUG (methionine) to AUA (isoleucine). A scintillation proximity assay for the incorporation of lysine into Ile2 tRNA was used to eliminate false positives in the initial screen resulting from detection artifacts as well as compounds competitive with the fluorescent label instead of ATP, and to measure inhibitor potencies against E. coli and Pseudomonas aeruginosa TilS isozymes. The tRNA(Ile) substrate for P. aeruginosa TilS was identified for the first time to enable these measurements. ATP-competitive binding of inhibitors was confirmed by one-dimensional ligand-observe nuclear magnetic resonance. A preliminary structure-activity relationship is shown for two inhibitor series.

  9. Evaluation of TILS for use as the orbiter landing NAVAID

    NASA Technical Reports Server (NTRS)

    Tate, J. M.

    1974-01-01

    An evaluation of the tactical instrument landing systems (TILS) for use in the orbiter autoland system was made. It was found that with certain modifications, the TILS can satisfy orbiter autoland requirements. These modifications, include (1) addition of DME equipment, (2) expansion of elevation coverage from 0-10 deg to 0-30 deg, and (3) expansion to redundant systems with associated ground monitors. Additional modifications that are not necessary to meet the orbiter requirements, but that can enhance performance margin are (1) tightening of elevation antenna beam width from 1.3 deg to 0.5 deg and (2) split site configuration to provide azimuth and range coverage through rollout.

  10. Telltale Tumor Infiltrating Lymphocytes (TIL) in Oral, Head & Neck Cancer

    PubMed Central

    Lei, Yu; Xie, Yuying; Tan, Yee Sun; Prince, Mark E.; Moyer, Jeffrey S.; Nör, Jacques; Wolf, Gregory T.

    2017-01-01

    Evidence gleaned from recent studies on the role of tumor-infiltrating lymphocytes (TILs) suggests that cancer is not only a genetic disease but also an immunologic disease. Head and Neck Squamous Cell Carcinoma (HNSCC) has been a significant model to study cancer cell-immune cell interactions. First, immune cell infiltration is an important feature of these tumors. Second, HNSCC frequently develops resistance to immunogenic cytotoxicity, which provides a window to decipher how tumors engage the immune system to establish immune tolerance. Finally, chemoradiation therapy, as a central modality for HNSCC treatment, has been shown to elicit immune activation. The presence of effector immune cells in the tumor microenvironment is often associated with superior clinical response to adjuvant therapy. On the other hand, an activated immune system, in addition to limiting tumor initiation and progression, could also exert selective pressure to promote the growth of less immunogenic tumors, as a pivotal immunoediting process. But it remains unclear how cancer cell signaling regulates tumor immunogenicity and how to mitigate HNSCC-potentiated TIL suppression. In this review, we will revisit the prognostic role of TILs in HNSCC, and collectively discuss how cancer cell machinery impacts upon the plasticity of TILs. PMID:27553942

  11. Expressions of CD8+TILs, PD-L1 and Foxp3+TILs in stage I NSCLC guiding adjuvant chemotherapy decisions

    PubMed Central

    Teng, Feifei; Meng, Xiangjiao; Wang, Xin; Yuan, Jupeng; Liu, Sujing; Mu, Dianbin; Zhu, Hui; Kong, Li; Yu, Jinming

    2016-01-01

    Purpose Currently, adjuvant chemotherapy is recommended for patients with high risk stage I non-small cell lung cancer (NSCLC). However, identifying high risk patients remains a challenge. This study aims to identify the patient cohorts more likely to benefit from adjuvant chemotherapy based on the tumor micro-immune environment. Results CD8+TILs significantly associated with disease-free survival (DFS) and overall survial (OS) (p=0.002; 0.040). Patients with high risk factors may also predict shorter DFS (P=0.056). When compared together, patients with high-CD8+TILs showed better DFS than patients with low-CD8+TILs, no matter their risk factors status. There's no correlation between PD-L1 expressions and survival. PD-L1 was highly expressed in men, squamous and well differentiated carcinoma. In addition, Foxp3+TILs alone didn't show any prognostic effects, but low-Foxp3/high-CD8+TILs were associated with prolonged DFS (p=0.031). Methods A total of 126 patients with surgically resected stage I NSCLC were included to perform immunohistochemistry of CD8+ tumor infiltrating lymphocytes (TILs), programmed death ligand-1(PD-L1) and forkhead box P3 (Foxp3)+TILs. Conclusion CD8+TILs are effective prognostic predictors. Patients with surgically resected stage I NSCLC showing low CD8+TILs could be considered for adjuvant chemotherapy, even if they have no high risk features. PMID:27602763

  12. Hearing in Drosophila Requires TilB, a Conserved Protein Associated With Ciliary Motility

    PubMed Central

    Kavlie, Ryan G.; Kernan, Maurice J.; Eberl, Daniel F.

    2010-01-01

    Cilia were present in the earliest eukaryotic ancestor and underlie many biological processes ranging from cell motility and propulsion of extracellular fluids to sensory physiology. We investigated the contribution of the touch insensitive larva B (tilB) gene to cilia function in Drosophila melanogaster. Mutants of tilB exhibit dysfunction in sperm flagella and ciliated dendrites of chordotonal organs that mediate hearing and larval touch sensitivity. Mutant sperm axonemes as well as sensory neuron dendrites of Johnston's organ, the fly's auditory organ, lack dynein arms. Through deficiency mapping and sequencing candidate genes, we identified tilB mutations in the annotated gene CG14620. A genomic CG14620 transgene rescued deafness and male sterility of tilB mutants. TilB is a 395-amino-acid protein with a conserved N-terminal leucine-rich repeat region at residues 16–164 and a coiled-coil domain at residues 171–191. A tilB-Gal4 transgene driving fluorescently tagged TilB proteins elicits cytoplasmic expression in embryonic chordotonal organs, in Johnston's organ, and in sperm flagella. TilB does not appear to affect tubulin polyglutamylation or polyglycylation. The phenotypes and expression of tilB indicate function in cilia construction or maintenance, but not in intraflagellar transport. This is also consistent with phylogenetic association of tilB homologs with presence of genes encoding axonemal dynein arm components. Further elucidation of tilB functional mechanisms will provide greater understanding of cilia function and will facilitate understanding ciliary diseases. PMID:20215474

  13. The Educational Thought of Cornelius Van Til: Philosophical Foundations of the Contemporary Christian School Movement.

    ERIC Educational Resources Information Center

    Maffet, Gregory J.; Dye, Charles M.

    This paper gives an account of the thoughts of Cornelius Van Til on the contemporary Christian school movement. An account of the historical development of Christian compromise is given, followed by a critique of the compromise among contemporary Christian educators. Van Til claims that any educational position which falls short of being founded…

  14. Adoptive TIL transfer in the adjuvant setting for melanoma: long-term patient survival.

    PubMed

    Khammari, Amir; Knol, Anne-Chantal; Nguyen, Jean-Michel; Bossard, Céline; Denis, Marc-Guillaume; Pandolfino, Marie-Christine; Quéreux, Gaëlle; Bercegeay, Sylvain; Dréno, Brigitte

    2014-01-01

    Two first analyses of our clinical trial on TIL as adjuvant therapy for melanoma were published in 2002 and 2007. We present here an update of the clinical results after a 17-year median followup. In this trial, disease-free patients were randomly assigned to receive either TIL/IL-2 or IL-2. The relapse-free survival (RFS) was the primary objective. Eighty-eight patients were enrolled. A new analysis performed in May 2013 did not show significant changes in RFS or OS duration. However, our first finding on the association between the number of invaded lymph nodes and TIL effectiveness was strengthened. The Cox model adjusted on this interaction showed for the first time a significant treatment effect when considering the overall population, both on the RFS and OS. Patients treated with TIL had a longer RFS (P = 0.023) or OS (P = 0.020). This study being with a very long followup (17 years), confirmed the association between TIL effectiveness and the number of invaded lymph nodes, indicating that a low tumor burden could be a crucial factor enhancing the curative effect of TIL in possible microscopic residual disease. Moreover, we confirmed that a prolonged survival was associated with the presence of specific TIL and a decrease in Foxp3 expression.

  15. Use of Tumor-infiltrating lymphocytes (TILs) to predict the treatment response to eribulin chemotherapy in breast cancer

    PubMed Central

    Kashiwagi, Shinichiro; Goto, Wataru; Takada, Koji; Takahashi, Katsuyuki; Noda, Satoru; Takashima, Tsutomu; Onoda, Naoyoshi; Tomita, Shuhei; Ohsawa, Masahiko; Hirakawa, Kosei

    2017-01-01

    Background Eribulin mesylate (eribulin) is currently indicated for treatment of locally advanced or metastatic breast cancer (MBC). It is a cytotoxic agent with unique mechanisms that suppress epithelial-mesenchymal transition (EMT) of cancer cells. On the other hand, Tumor-infiltrating lymphocytes (TILs), which are considered indicators of immune response monitoring, have been reported as prognostic factors and predictors of therapeutic efficacy. We thought that eribulin, which has an EMT-inhibiting mechanism, may produce an antitumor effect by improving the immune microenvironment, and in this study investigated the effects of breast cancer eribulin chemotherapy on the immune microenvironment with TILs as a marker. Methods TILs was evaluated in 52 patients with MBC who underwent chemotherapy with eribulin. The correlation between TILs evaluated according to the standard method, and prognosis, including the efficacy of eribulin chemotherapy, was investigated retrospectively. Results Of the 52 MBC patients, 29 (55.8%) were in the high TILs group and 23 (44.2%) were in the low TILs group. The high TILs group included significantly more triple-negative breast cancer (TNBC) (p = 0.008) than the low TILs group. In an analysis of outcomes, TNBC patients in the high TILs group had significantly longer disease-free survival than TNBC patients in the low TILs group (p = 0.033, log-rank), but no significant differences were seen in all breast cancer patients (p = 0.489, log-rank) or in non-TNBC patients (p = 0.878, log-rank). In a multivariate analysis of recurrence in TNBC patients, being in the high TILs group was again an independent factor for a good outcome (p = 0.031, HR = 0.063). Conclusion The results of this study suggest that TILs may be useful as a predictive marker of the therapeutic effect of eribulin chemotherapy in TNBC. PMID:28166544

  16. [The life of Choe Ung-sok: with a focus on his design for and role in the health care system immediately after the liberation].

    PubMed

    Shin, Young-Joen; Kim, Jinhyouk

    2014-12-01

    Born in Pyongyang in 1914, Choe Ung-sok was a physician who lived through the Japanese colonial era (1910-1945), rule by the United States Army Military Government in Korea (USAMGIK; 1945-1948), and national division (1948). Influenced by socialism and social hygiene/social medicine during his studies in Japan, he played the role of representing the socialist camp in the discussions related to the construction of a heath care system immediately following the Liberation (1945). His key arguments were: first, the nationalization of the medical system and the implementation of nationwide programs to eradicate diseases; second, the provision of free medical services through the expansion of social insurance; third, the reeducation of the medical personnel; fourth, the provision of social sciences education to the medical personnel and the reorganization of medicine into preventive medicine; fifth, the nationalization of pharmaceutics; sixth, the laborers' establishment of autonomous medical organs (affordable clinics, medical consumers' unions through cooperatives); and seventh, the reduction of work hours to 6-8 hours, technical improvement, respite from research, and guarantee of economic life for the medical personnel. Influenced by the medical systems of the Soviet Union and Japan, such arguments stood in opposition to the right wing's plan for the construction of a relatively passive health care system at the time but, in the end, failed to be realized in southern part of Korea under the USAMGIK. Subsequently, he defected to northern part of Korea and came to participate in the task of constructing North Korea's health care system. Choe's life and design for a health care system provide examples through which one can confirm the nature of social hygiene/social medicine both during the Japanese colonial era and before and after the Liberation and the contents of the design related to a health care system as held by the socialist faction. In addition, they show that

  17. Aircraft borne combined measurements of the Fukushima radionuclide Xe-133 and fossil fuel combustion generated pollutants in the TIL - Implications for Cyclone induced lift and TIL physical-chemical processes

    NASA Astrophysics Data System (ADS)

    Arnold, Frank; Schlager, Hans; Simgen, Hardy; Aufmhoff, Heinfried; Baumann, Robert; Lindemann, Sigfried; Rauch, Ludwig; Kaether, Frank; Pirjolla, Liisa; Schumann, Ulrich

    2013-04-01

    The radionuclide Xe-133, released by the March 2011 nuclear disaster at Fukushima/Daiichi (hereafter FD), represents an ideal tracer for atmospheric transport. We report the, to our best knowledge, only aircraft borne measurements of FD Xe-133 in the Tropopause Inversion Layer (TIL), indicating rapid lift of Xe-133 rich planetary boundary layer air to the TIL. On the same research aircraft (FALCON), we have also conducted on-line measurements of fossil fuel combustion generated pollutant gases (SO2, NOx, HNO3,NOy), which were found to have increased concentrations in the TIL. In addition, we have conducted supporting model simulations of transport, chemical processes, and aerosol processes. Our investigations reveal a potentially important influence of East-Asian cyclone induced pollutants transport to the TIL, particularly influencing aerosol formation in the TIL.

  18. Tumor infiltrating lymphocytes (TILs) improve prognosis in patients with triple negative breast cancer (TNBC).

    PubMed

    Adams, Sylvia; Goldstein, Lori J; Sparano, Joseph A; Demaria, Sandra; Badve, Sunil S

    2015-09-01

    Upon analysis of archived primary tumors of 482 patients with triple negative breast cancer (TNBC) enrolled in two randomized Phase III adjuvant chemotherapy trials, we have found that tumor infiltrating lymphocytes (TILs), as assessed and quantified by hematoxylin and eosin (H&E) staining are a robust and independent predictor of disease-free survival (DFS), distant recurrence-free interval (DRFI) and overall survival (OS).(1) Our findings provide confirmation of results observed in TNBC in a European adjuvant chemotherapy dataset and therefore elevate TILs as prognostic biomarker for operable TNBC to level I evidence.

  19. Evaluation of particulate filtering respirators using inward leakage (IL) or total inward leakage (TIL) testing--Korean experience.

    PubMed

    Han, Don-Hee; Lee, Jinheon

    2005-10-01

    Korean certification regulation for particulate filtering respirators requires inward leakage (IL) or total inward leakage (TIL) testing according to European Standard EN 13274-1, and the standard levels of compliance are similar to those of the European Standard. This study was conducted to evaluate particulate filtering respirators being commercially used in the Korean market using an IL or TIL test and the validity of standard level in Korea. Three half masks and 10 filtering facepieces (two top class, four 1st class and four 2nd class)-a total of 13 brand name respirators-were selected for the test with panels of 10 subjects. Each subject was classified with nine facial dimension grid squares in accordance with face length and lip length. IL or TIL testing was conducted at the laboratory of the 3M Innovation Center in which the experimental instruments and systems were established in compliance with European standards. The testing procedure followed EN 13274-1 (2001). As expected, leakages of half masks were less than those of filtering facepieces and the latter were significantly different among brands. TILs of the 1st class filtering facepieces were found to be much more than those of the 2nd class and the result may cause a wearer to get confused when selecting a mask. The main route leakage for filtering facepieces may not be the filter medium but the face seal. Therefore, it is necessary to develop well-fitting filtering facepieces for Koreans. Because leakages were significantly different for different facial dimensions, a defined test panel for IL or TIL testing according to country or race should be developed. A more precise method to demonstrate fit, for example, fit testing such as in the US regulations, will be needed before IL or TIL testing or when selecting a respirator. Another finding implies that geometric mean of five exercises for IL or TIL may be better than arithmetic mean to establish a standard individual subject mean.

  20. The 1-min Screening Test for Reading Problems in College Students: Psychometric Properties of the 1-min TIL.

    PubMed

    Fernandes, Tânia; Araújo, Susana; Sucena, Ana; Reis, Alexandra; Castro, São Luís

    2017-02-01

    Reading is a central cognitive domain, but little research has been devoted to standardized tests for adults. We, thus, examined the psychometric properties of the 1-min version of Teste de Idade de Leitura (Reading Age Test; 1-min TIL), the Portuguese version of Lobrot L3 test, in three experiments with college students: typical readers in Experiment 1A and B, dyslexic readers and chronological age controls in Experiment 2. In Experiment 1A, test-retest reliability and convergent validity were evaluated in 185 students. Reliability was >.70, and phonological decoding underpinned 1-min TIL. In Experiment 1B, internal consistency was assessed by presenting two 45-s versions of the test to 19 students, and performance in these versions was significantly associated (r = .78). In Experiment 2, construct validity, criterion validity and clinical utility of 1-min TIL were investigated. A multiple regression analysis corroborated construct validity; both phonological decoding and listening comprehension were reliable predictors of 1-min TIL scores. Logistic regression and receiver operating characteristics analyses revealed the high accuracy of this test in distinguishing dyslexic from typical readers. Therefore, the 1-min TIL, which assesses reading comprehension and potential reading difficulties in college students, has the necessary psychometric properties to become a useful screening instrument in neuropsychological assessment and research. Copyright © 2017 John Wiley & Sons, Ltd.

  1. TIL therapy broadens the tumor-reactive CD8(+) T cell compartment in melanoma patients.

    PubMed

    Kvistborg, Pia; Shu, Chengyi Jenny; Heemskerk, Bianca; Fankhauser, Manuel; Thrue, Charlotte Albæk; Toebes, Mireille; van Rooij, Nienke; Linnemann, Carsten; van Buuren, Marit M; Urbanus, Jos H M; Beltman, Joost B; Thor Straten, Per; Li, Yong F; Robbins, Paul F; Besser, Michal J; Schachter, Jacob; Kenter, Gemma G; Dudley, Mark E; Rosenberg, Steven A; Haanen, John B A G; Hadrup, Sine Reker; Schumacher, Ton N M

    2012-07-01

    There is strong evidence that both adoptive T cell transfer and T cell checkpoint blockade can lead to regression of human melanoma. However, little data are available on the effect of these cancer therapies on the tumor-reactive T cell compartment. To address this issue we have profiled therapy-induced T cell reactivity against a panel of 145 melanoma-associated CD8(+) T cell epitopes. Using this approach, we demonstrate that individual tumor-infiltrating lymphocyte cell products from melanoma patients contain unique patterns of reactivity against shared melanoma-associated antigens, and that the combined magnitude of these responses is surprisingly low. Importantly, TIL therapy increases the breadth of the tumor-reactive T cell compartment in vivo, and T cell reactivity observed post-therapy can almost in full be explained by the reactivity observed within the matched cell product. These results establish the value of high-throughput monitoring for the analysis of immuno-active therapeutics and suggest that the clinical efficacy of TIL therapy can be enhanced by the preparation of more defined tumor-reactive T cell products.

  2. TIL therapy broadens the tumor-reactive CD8+ T cell compartment in melanoma patients

    PubMed Central

    Kvistborg, Pia; Shu, Chengyi Jenny; Heemskerk, Bianca; Fankhauser, Manuel; Thrue, Charlotte Albæk; Toebes, Mireille; van Rooij, Nienke; Linnemann, Carsten; van Buuren, Marit M.; Urbanus, Jos H.M.; Beltman, Joost B.; thor Straten, Per; Li, Yong F.; Robbins, Paul F.; Besser, Michal J.; Schachter, Jacob; Kenter, Gemma G.; Dudley, Mark E.; Rosenberg, Steven A.; Haanen, John B.A.G.; Hadrup, Sine Reker; Schumacher, Ton N.M.

    2012-01-01

    There is strong evidence that both adoptive T cell transfer and T cell checkpoint blockade can lead to regression of human melanoma. However, little data are available on the effect of these cancer therapies on the tumor-reactive T cell compartment. To address this issue we have profiled therapy-induced T cell reactivity against a panel of 145 melanoma-associated CD8+ T cell epitopes. Using this approach, we demonstrate that individual tumor-infiltrating lymphocyte cell products from melanoma patients contain unique patterns of reactivity against shared melanoma-associated antigens, and that the combined magnitude of these responses is surprisingly low. Importantly, TIL therapy increases the breadth of the tumor-reactive T cell compartment in vivo, and T cell reactivity observed post-therapy can almost in full be explained by the reactivity observed within the matched cell product. These results establish the value of high-throughput monitoring for the analysis of immuno-active therapeutics and suggest that the clinical efficacy of TIL therapy can be enhanced by the preparation of more defined tumor-reactive T cell products. PMID:22754759

  3. Gene expression analysis of TIL rich HPV-driven head and neck tumors reveals a distinct B-cell signature when compared to HPV independent tumors

    PubMed Central

    Savelyeva, Natalia; McCann, Katy J.; Singh, Divya; Jones, Terry; Peel, Lailah; Breen, Michael S.; Ward, Matthew; Martin, Eva Garrido

    2016-01-01

    Human papilloma virus (HPV)-associated head and neck squamous cell carcinoma (HNSCC) has a better prognosis than it's HPV negative (HPV(−)) counterpart. This may be due to the higher numbers of tumor-infiltrating lymphocytes (TILs) in HPV positive (HPV(+)) tumors. RNA-Sequencing (RNA-Seq) was used to evaluate whether the differences in clinical behaviour simply reflect a numerical difference in TILs or whether there is a fundamental behavioural difference between TILs in these two settings. Thirty-nine HNSCC tumors were scored for TIL density by immunohistochemistry. After the removal of 16 TILlow tumors, RNA-Seq analysis was performed on 23 TILhigh/med tumors (HPV(+) n=10 and HPV(−) n=13). Using EdgeR, differentially expressed genes (DEG) were identified. Immune subset analysis was performed using Functional Analysis of Individual RNA-Seq/ Microarray Expression (FAIME) and immune gene RNA transcript count analysis. In total, 1,634 DEGs were identified, with a dominant immune signature observed in HPV(+) tumors. After normalizing the expression profiles to account for differences in B- and T-cell number, 437 significantly DEGs remained. A B-cell associated signature distinguished HPV(+) from HPV(−) tumors, and included the DEGs CD200, GGA2, ADAM28, STAG3, SPIB, VCAM1, BCL2 and ICOSLG; the immune signal relative to T-cells was qualitatively similar between TILs of both tumor cohorts. Our findings were validated and confirmed in two independent cohorts using TCGA data and tumor-infiltrating B-cells from additional HPV(+) HNSCC patients. A B-cell associated signal segregated tumors relative to HPV status. Our data suggests that the role of B-cells in the adaptive immune response to HPV(+) HNSCC requires re-assessment. PMID:27462861

  4. Gene expression analysis of TIL rich HPV-driven head and neck tumors reveals a distinct B-cell signature when compared to HPV independent tumors.

    PubMed

    Wood, Oliver; Woo, Jeongmin; Seumois, Gregory; Savelyeva, Natalia; McCann, Katy J; Singh, Divya; Jones, Terry; Peel, Lailah; Breen, Michael S; Ward, Matthew; Garrido Martin, Eva; Sanchez-Elsner, Tilman; Thomas, Gareth; Vijayanand, Pandurangan; Woelk, Christopher H; King, Emma; Ottensmeier, Christian

    2016-08-30

    Human papilloma virus (HPV)-associated head and neck squamous cell carcinoma (HNSCC) has a better prognosis than it's HPV negative (HPV(-)) counterpart. This may be due to the higher numbers of tumor-infiltrating lymphocytes (TILs) in HPV positive (HPV(+)) tumors. RNA-Sequencing (RNA-Seq) was used to evaluate whether the differences in clinical behaviour simply reflect a numerical difference in TILs or whether there is a fundamental behavioural difference between TILs in these two settings. Thirty-nine HNSCC tumors were scored for TIL density by immunohistochemistry. After the removal of 16 TILlow tumors, RNA-Seq analysis was performed on 23 TILhigh/med tumors (HPV(+) n=10 and HPV(-) n=13). Using EdgeR, differentially expressed genes (DEG) were identified. Immune subset analysis was performed using Functional Analysis of Individual RNA-Seq/ Microarray Expression (FAIME) and immune gene RNA transcript count analysis. In total, 1,634 DEGs were identified, with a dominant immune signature observed in HPV(+) tumors. After normalizing the expression profiles to account for differences in B- and T-cell number, 437 significantly DEGs remained. A B-cell associated signature distinguished HPV(+) from HPV(-) tumors, and included the DEGs CD200, GGA2, ADAM28, STAG3, SPIB, VCAM1, BCL2 and ICOSLG; the immune signal relative to T-cells was qualitatively similar between TILs of both tumor cohorts. Our findings were validated and confirmed in two independent cohorts using TCGA data and tumor-infiltrating B-cells from additional HPV(+) HNSCC patients. A B-cell associated signal segregated tumors relative to HPV status. Our data suggests that the role of B-cells in the adaptive immune response to HPV(+) HNSCC requires re-assessment.

  5. Deep Sequencing of T-Cell Receptor DNA as a biomarker of clonally expanded TILs in breast cancer after immunotherapy

    PubMed Central

    Page, David B.; Yuan, Jianda; Redmond, David; Wen, Y Hanna; Durack, Jeremy C.; Emerson, Ryan; Solomon, Stephen; Dong, Zhiwan; Wong, Phillip; Comstock, Christopher; Diab, Adi; Sung, Janice; Maybody, Majid; Morris, Elizabeth; Brogi, Edi; Morrow, Monica; Sacchini, Virgilio; Elemento, Olivier; Robins, Harlan; Patil, Sujata; Allison, James P.; Wolchok, Jedd D.; Hudis, Clifford; Norton, Larry; McArthur, Heather

    2016-01-01

    In early stage breast cancer, the degree of tumor-infiltrating lymphocytes (TILs) predicts response to chemotherapy and overall survival. Combination immunotherapy with immune checkpoint antibody plus tumor cryoablation can induce lymphocytic infiltrates and improve survival in mice. We used T-cell receptor (TCR) DNA sequencing to evaluate both the effect of cryo-immunotherapy in humans and the feasibility of TCR sequencing in early-stage breast cancer. In a pilot clinical trial, 18 women with early-stage breast cancer were treated preoperatively with cryoablation, single-dose anti-CTLA-4 (ipilimumab), or cryoablation + ipilimumab. TCRs within serially collected peripheral blood and tumor tissue were sequenced. In baseline tumor tissues, T-cell density as measured by TCR sequencing correlated with TIL scores obtained by hematoxylin and eosin (H&E) staining. However, tumors with little or no lymphocytes by H&E contained up to 3.6 × 106 TCR DNA sequences, highlighting the sensitivity of the ImmunoSEQ platform. In this dataset, ipilimumab increased intratumoral T-cell density over time, whereas cryoablation ± ipilimumab diversified and remodeled the intratumoral T-cell clonal repertoire. Compared to monotherapy, cryoablation plus ipilimumab was associated with numerically greater numbers of peripheral blood and intratumoral T-cell clones expanding robustly following therapy. In conclusion, TCR sequencing correlates with H&E lymphocyte scoring, and provides additional information on clonal diversity. These findings support further study of the use of TCR sequencing as a biomarker for T cell responses to therapy and for the study of cryo-immunotherapy in early-stage breast cancer. PMID:27587469

  6. Deep Sequencing of T-cell Receptor DNA as a Biomarker of Clonally Expanded TILs in Breast Cancer after Immunotherapy.

    PubMed

    Page, David B; Yuan, Jianda; Redmond, David; Wen, Y Hanna; Durack, Jeremy C; Emerson, Ryan; Solomon, Stephen; Dong, Zhiwan; Wong, Phillip; Comstock, Christopher; Diab, Adi; Sung, Janice; Maybody, Majid; Morris, Elizabeth; Brogi, Edi; Morrow, Monica; Sacchini, Virgilio; Elemento, Olivier; Robins, Harlan; Patil, Sujata; Allison, James P; Wolchok, Jedd D; Hudis, Clifford; Norton, Larry; McArthur, Heather L

    2016-10-01

    In early-stage breast cancer, the degree of tumor-infiltrating lymphocytes (TIL) predicts response to chemotherapy and overall survival. Combination immunotherapy with immune checkpoint antibody plus tumor cryoablation can induce lymphocytic infiltrates and improve survival in mice. We used T-cell receptor (TCR) DNA sequencing to evaluate both the effect of cryoimmunotherapy in humans and the feasibility of TCR sequencing in early-stage breast cancer. In a pilot clinical trial, 18 women with early-stage breast cancer were treated preoperatively with cryoablation, single-dose anti-CTLA-4 (ipilimumab), or cryoablation + ipilimumab. TCRs within serially collected peripheral blood and tumor tissue were sequenced. In baseline tumor tissues, T-cell density as measured by TCR sequencing correlated with TIL scores obtained by hematoxylin and eosin (H&E) staining. However, tumors with little or no lymphocytes by H&E contained up to 3.6 × 10(6) TCR DNA sequences, highlighting the sensitivity of the ImmunoSEQ platform. In this dataset, ipilimumab increased intratumoral T-cell density over time, whereas cryoablation ± ipilimumab diversified and remodeled the intratumoral T-cell clonal repertoire. Compared with monotherapy, cryoablation plus ipilimumab was associated with numerically greater numbers of peripheral blood and intratumoral T-cell clones expanding robustly following therapy. In conclusion, TCR sequencing correlates with H&E lymphocyte scoring and provides additional information on clonal diversity. These findings support further study of the use of TCR sequencing as a biomarker for T-cell responses to therapy and for the study of cryoimmunotherapy in early-stage breast cancer. Cancer Immunol Res; 4(10); 835-44. ©2016 AACR.

  7. TIL-type protease inhibitors may be used as targeted resistance factors to enhance silkworm defenses against invasive fungi.

    PubMed

    Li, Youshan; Zhao, Ping; Liu, Huawei; Guo, Xiaomeng; He, Huawei; Zhu, Rui; Xiang, Zhonghuai; Xia, Qingyou

    2015-02-01

    Entomopathogenic fungi penetrate the insect cuticle using their abundant hydrolases. These hydrolases, which include cuticle-degrading proteases and chitinases, are important virulence factors. Our recent findings suggest that many serine protease inhibitors, especially TIL-type protease inhibitors, are involved in insect resistance to pathogenic microorganisms. To clarify the molecular mechanism underlying this resistance to entomopathogenic fungi and identify novel genes to improve the silkworm antifungal capacity, we conducted an in-depth study of serine protease inhibitors. Here, we cloned and expressed a novel silkworm TIL-type protease inhibitor, BmSPI39. In activity assays, BmSPI39 potently inhibited the virulence protease CDEP-1 of Beauveria bassiana, suggesting that it might suppress the fungal penetration of the silkworm integument by inhibiting the cuticle-degrading proteases secreted by the fungus. Phenol oxidase activation studies showed that melanization is involved in the insect immune response to fungal invasion, and that fungus-induced excessive melanization is suppressed by BmSPI39 by inhibiting the fungal cuticle-degrading proteases. To better understand the mechanism involved in the inhibition of fungal virulence by protease inhibitors, their effects on the germination of B. bassiana conidia was examined. BmSPI38 and BmSPI39 significantly inhibited the germination of B. bassiana conidia. Survival assays showed that BmSPI38 and BmSPI39 markedly improved the survival rates of silkworms, and can therefore be used as targeted resistance proteins in the silkworm. These results provided new insight into the molecular mechanisms whereby insect protease inhibitors confer resistance against entomopathogenic fungi, suggesting their potential application in medicinal or agricultural fields.

  8. CD3+, CD4+ & CD8+ tumour infiltrating lymphocytes (TILs) are predictors of favourable survival outcome in infiltrating ductal carcinoma of breast

    PubMed Central

    Rathore, Ankita Singh; Kumar, Sandeep; Konwar, Rituraj; Makker, Annu; Negi, M.P.S.; Goel, Madhu Mati

    2014-01-01

    Background & objectives: Tumour infiltrating lymphocytes (TILs) represent the host immune response against cancer cells associated with good or bad prognosis in different tumour types. This study was undertaken to evaluate the significance of CD3+, CD4+ and CD8+ TILs in breast cancer tissues in relation to clinico-pathological variables and survival outcome. Methods: Immunohistochemistry (IHC) was performed with antibodies against CD3, CD4 and CD8 antigens on formalin-fixed paraffin-embedded tissue sections of 150 breast cancer patients. Intratumoural and stromal TIL counting was performed semiquantitatively. Results: The higher CD3+, CD4+ and CD8+ intratumoural and stromal counts showed independent and direct association with good prognosis. The prognostic predictor value of intratumoural counts was higher than stromal counts. The independent associations of intratumoural and stromal counts became more prominent when adjusted with stage and grade, respectively. Among intratumoural counts, the high (++/+++) CD4+ count (OR=3.85, 95% CI=3.28-16.71, P<0.001) showed the highest survival followed by CD3+ (OR=2.70, 95% CI=1.76-8.30, P=0.001) and CD8+ (OR=2.58, 95% CI=1.55-5.86, P=0.001) the least when compared to respective low (+) counts. In contrast, among stromal counts, the high CD8+ count (OR=3.13, 95% CI=2.20-9.57, P<0.001) showed the highest survival followed by CD4+ (OR=3.02, 95% CI=2.07-8.89, P<0.001) and CD3+ (OR=2.45, 95% CI=1.53-6.73, P=0.002) the least. Interpretation & conclusions: Our results suggest that intratumoural CD4+ and stromal CD8+ counts by immunohistochemistry may serve as an independent prognosticator for favourable outcome in breast cancer. PMID:25366203

  9. 'Til the Needle Breaks

    ERIC Educational Resources Information Center

    Hawkins, B. Denise

    2011-01-01

    After more than half a century, the music of Motown not only thrives, it transcends generations. The iconic sound of Motown has led a handful of scholars to write, teach, lecture and share the music, history and business of Motown on their campuses. In its golden age, from 1959 to 1972, the sound Berry Gordy pioneered at Motown Records in Detroit…

  10. Erschließung eines Marmorkarstvorkommens als mitteltiefer Erdwärmesondenspeicher im Tuxertal, Tirol

    NASA Astrophysics Data System (ADS)

    Sass, Ingo; Heldmann, Claus-Dieter; Lehr, Clemens

    2016-06-01

    Borehole heat exchangers can be economically beneficial for meeting heating and cooling demands of houses or buildings. In karst aquifers development of thermal storage and exchange systems may be problematic in terms of groundwater protection and storage design, due to possibly high groundwater velocities. The new development of the Hochstegen marble unit in the Tux Valley (Zillertal, Austria) was designed in two stages for the requested cooling and heating demands. An enhanced geothermal response test was completed using optical frequency domain reflectometry in an exploration drillhole. Additional studies focussing on local geology and hydrology were also conducted. Geothermal parameters obtained at precise depths allowed differentiating between conductive and convective heat flow and were correlated with the lithostratigraphically-conditioned karst characteristics. The borehole heat exchanger field was developed with nine 400 m deep dual U-shaped tube probes in 2013 for 1 GWh/a extraction and 400 MWh/a induction. Along with borehole geophysics and geothermal response tests, the study has provided relevant geothermal data for improving storage design and exploration.

  11. Erkundung und Beweissicherung für eine geothermale Erschließung eines Alpinen Karstaquifers im Tuxertal, Österreich

    NASA Astrophysics Data System (ADS)

    Sass, Ingo; Heldmann, Claus-Dieter; Schäffer, Rafael

    2016-06-01

    Karst aquifers may on one hand improve the efficiency of geothermal systems due to increased permeabilities, but on the other hand, high groundwater velocities can reduce the efficiency of the underground heat storage capacity. The marble karst aquifer of the Hochstegen formation was explored and developed for the first time as an intermediate-depth geothermal energy storage system at Finkenberg, Tux valley (Tyrol, Austria). Geological field studies and a spring monitoring program for the project revealed characteristic hydro-chemical signatures related to the catchments in specific tectonic units depending on their lithology. Observations showed that the catchment area of the Hochstegen formation karst aquifer extends up to 2650 m a.s.l. southwest of Finkenberg. In the boreholes, karstification was detected to 400 m below surface (Sass et al., 2016). A monitoring program involving seven springs downgradient of the boreholes has shown that the geothermal project has had no long-term impact on groundwater quality.

  12. Registration of TIL:383.13, TIL:625 and TIL:634, three long grain tropical Japonica Rice (Oryza sativa L.) germplasm lines containing novel Indica Alleles that increase tiller production and grain yield

    Technology Transfer Automated Retrieval System (TEKTRAN)

    These three breeding lines were from a set of 123 progeny lines that were released by the USDA-ARS in 2012 as a mapping population. Chromosomal regions containing genes for increased tiller number under greenhouse conditions were subsequently identified in this population. We used the molecular an...

  13. The Repeal of Section 28: It Ain't over 'til It's over

    ERIC Educational Resources Information Center

    Greenland, Katy; Nunney, Rosalind

    2008-01-01

    Section 28 (part of the Local Government Act of 1988) was a notorious piece of legislation that sought to prevent local education authorities in the UK from "promoting homosexuality". The effect of Section 28 was to create uncertainty and fear among teachers as to what was (and what was not) permitted in schools. Over time practitioners…

  14. 'Keep complaining til someone listens': Exchanges of tacit healthcare knowledge in online illness communities.

    PubMed

    Foster, Drew

    2016-10-01

    This article examines online exchanges of advice and knowledge among patients. It draws a distinction between explicit healthcare knowledge (i.e., facts about symptoms and treatments) and tacit healthcare knowledge (i.e., know-how about navigating the healthcare system). Based on analysis of message board interactions at a prominent online illness community, I find that patients routinely encourage one another to exercise agency strategically in clinical encounters by honing specific interactional skills. I isolate three major techniques that are advocated within the community (affect regulation, information management, and treatment persistence) and frame them as discrete examples of tacit healthcare knowledge. I argue that tacit healthcare knowledge constitutes a potentially potent source of empowerment for patients that can help them to receive their desired form of care from the health system and to negotiate relationships with medical professionals and institutions. I conclude by discussing how the concept of tacit healthcare knowledge further clarifies the wide variety of lay knowledge exchanged among patients online.

  15. William Van Til and the Nashville Story: Curriculum, Supervision, and Civil Rights

    ERIC Educational Resources Information Center

    Perlstein, Daniel

    2004-01-01

    Massive white resistance to the Supreme Court's 1954 Brown decision encouraged educational administrators and leaders who supported school integration to ally themselves with progressive civic activists. Desegregation thus catalyzed the development of a new, more openly political vision of educational leadership. If schools of education were to…

  16. Clinical relevance of host immunity in breast cancer: from TILs to the clinic.

    PubMed

    Savas, Peter; Salgado, Roberto; Denkert, Carsten; Sotiriou, Christos; Darcy, Phillip K; Smyth, Mark J; Loi, Sherene

    2016-04-01

    The clinical relevance of the host immune system in breast cancer has long been unexplored. Studies developed over the past decade have highlighted the biological heterogeneity of breast cancer, prompting researchers to investigate whether the role of the immune system in this malignancy is similar across different molecular subtypes of the disease. The presence of high levels of lymphocytic infiltration has been consistently associated with a more-favourable prognosis in patients with early stage triple-negative and HER2-positive breast cancer. These infiltrates seem to reflect favourable host antitumour immune responses, suggesting that immune activation is important for improving survival outcomes. In this Review, we discuss the composition of the immune infiltrates observed in breast cancers, as well as data supporting the clinical relevance of host antitumour immunity, as represented by lymphocytic infiltration, and how this biomarker could be used in the clinical setting. We also discuss the rationale for enhancing immunity in breast cancer, including early data on the efficacy of T-cell checkpoint inhibition in this setting.

  17. 'Til death parts us: women’s domestic partnerships in eighteenth-century Brittany.

    PubMed

    Locklin, Nancy

    2011-01-01

    This article investigates the legal provision for two adult, unmarried women to create a “perpetual society” with one another found in the customary code of 1725 for the French province of Brittany. This arrangement allowed women who shared a household to designate one another as primary heir and to protect their community property from the claims of others. Evidence of this arrangement demonstrates that single women in some places had options outside of marriage and the convent. The contracts filed by the women also reveal the extent to which this arrangement went beyond considerations of property to express both affection and loyalty. Available to siblings as well as to pairs of unrelated women, this union is likely not the equivalent of same-sex marriage. It does however broaden our knowledge of the meaning of marriage, partnership, and kin in early modern Europe.

  18. ''Fake It 'Til You Make It'': Why Community College Students' Aspirations ''Hold Steady''

    ERIC Educational Resources Information Center

    Nielsen, Kelly

    2015-01-01

    Sociologists of education have explored the relationship between students' postsecondary aspirations and their propensity to get "cooled out" in community colleges. However, researchers have directed little attention to students whose aspirations remain stable over long periods of time or to the different roles that college degree goals…

  19. Twist `til we tear the house down: How Clifford solved the universe in 1870

    NASA Astrophysics Data System (ADS)

    Beichler, James

    2010-02-01

    It is commonly believed that the first hyperspace theories in physics were developed in the early twentieth century - Kaluza's five-dimensional extension of relativity is the best known, but this is untrue. It is also commonly believed that W.K. Clifford `speculated' on a higher space in 1870, had no followers and never published his theory (if he even had one). Nothing could be further from the historical truth. As early as 1869, Clifford, his followers and students began to develop a physical theory of matter based on a three-dimensional space curved in four dimensions. Clifford began to publish his theory, but modern researchers have failed to recognize his theoretical work because they look for something like Einstein's theory even though Clifford developed an electromagnetic theory. Clifford may not have directly influenced Einstein's relativity, but he made plausible arguments for the reality of space curvature, rendering the rapid acceptance of Einstein's concept of curved space-time more plausible. Clifford's work is either largely ignored by historians, scientists and other scholars or considered irrelevant because the early work on hyperspaces has been associated with ether theories that were abandoned, utilized quaternion algebras that were replaced by vectors and tensors, and was unfortunately associated with spiritualism. )

  20. Medical treatment choices for patients affected by advanced NSCLC in routine clinical practice: results from the Italian observational "SUN" (Survey on the lUng cancer maNagement) study.

    PubMed

    Gridelli, Cesare; Ardizzoni, Andrea; Barni, Sandro; Crinò, Lucio; Caprioli, Alberto; Piazza, Elena; Lorusso, Vito; Barbera, Santi; Zilembo, Nicoletta; Gebbia, Vittorio; Adamo, Vincenzo; Pela, Riccardo; Marangolo, Maurizio; Morena, Raffaella; Filippelli, Gianfranco; Buscarino, Calogero; Alabiso, Oscar; Maione, Paolo; Venturino, Paola; De Marinis, Filippo

    2011-12-01

    Lung cancer is the most common cancer in the world today, in terms of both incidence and mortality. Non-small cell lung cancer (NSCLC) accounts for about 85% of all lung cancers, and the majority of people diagnosed with NSCLC have locally advanced or metastatic disease. Treatment algorithms have rapidly changed in the last 10 years because of the introduction of new chemotherapeutic and targeted agents in clinical practice. SUN is a 1-year longitudinal observational multicenter study that has consecutively enrolled patients affected by stage IIIB or IV NSCLC with the aim to describe the pattern of care and evolving approaches in the treatment of advanced NSCLC. 987 consecutive NSCLC patients were enrolled between January 2007 and March 2008 at the 74 participating centers throughout Italy and a 12-month follow-up was performed. Cyto-histological diagnosis was performed mainly by broncoscopy with only 24% by CT-scan guided fine-needle aspiration biopsy. 91.4% of the patients received a first-line medical treatment and 8.6% supportive care only. Median age of patients receiving first-line treatment was 66 years. First-line chemotherapy consisted of a single agent in 20% of patients and combination chemotherapy in 80%. The most frequently used chemotherapy regimens were cisplatin plus gemcitabine and carboplatin plus gemcitabine. Median survival of patients receiving first-line chemotherapy was 9.1 months. 32% percent of patients received a second-line treatment that consisted of chemotherapy in 71% of cases and erlotinib in 29%. Overall third-line treatment was given to 7.3% of patients. These results showed a pattern of care for advanced NSCLC that reflects the current clinical practice in Italy at the study time with a high adherence to the International guidelines by the Italian Oncologists.

  1. Monte Carlo and deterministic simulations of activation ratio experiments for 238U(n,f), 238U(n,g) and 238U(n,2n) in the Big Ten benchmark critical assembly

    SciTech Connect

    Descalle, M; Clouse, C; Pruet, J

    2009-07-28

    The authors have compared calculations of critical assembly activation ratios using 3 different Monte Carlo codes and one deterministic code. There is excellent agreement. Discrepancies between the different Monte Carlo codes are the 1-2% level. Notably, the deterministic calculations with 87 groups are also in good agreement with the continuous energy Monte Carlo results. The three codes underestimate the {sup 238}U(n,f) reaction, suggesting that there is room for improvement in the evaluation, or in the evaluations of other reactions influencing the spectrum in BigTen. Until statistical uncertainties are implemented in Mercury, they strongly advise long runs to guarantee sufficient convergence of the flux at high energies, and they strongly encourage comparing Mercury results to a well-developed and documented code such as MCNP5 and/or COG. It may be that ENDL2008 will be available for use in COG within a year. Finally, it may be worthwhile to add a 'standard' reaction rate tally similar to those implemented in COG and MCNP5, if the goal is to expand the central fission and activation ratios simulations to include isotopes that are not part of the specifications for the assembly material composition.

  2. Physical and functional interaction of human nuclear uracil-DNA glycosylase with proliferating cell nuclear antigen☆

    PubMed Central

    Ko, Rinkei; Bennett, Samuel E.

    2011-01-01

    Uracil residues arise in DNA by the misincorporation of dUMP in place of dTMP during DNA replication or by the deamination of cytosine in DNA. Uracil-DNA glycosylase initiates DNA base excision repair of uracil residues by catalyzing the hydrolysis of the N-glycosylic bond linking the uracil base to deoxyribose. In human cells, the nuclear form of uracil-DNA glycosylase (UNG2) contains a conserved PCNA-binding motif located at the N-terminus that has been implicated experimentally in binding PCNA. Here we use purified preparations of UNG2 and PCNA to demonstrate that UNG2 physically associates with PCNA. UNG2 co-eluted with PCNA during size exclusion chromatography and bound to a PCNA affinity column. Association of UNG2 with PCNA was abolished by the addition of 100 mM NaCl, and significantly decreased in the presence of 10 mM MgCl2. The functional significance of the UNG2·PCNA association was demonstrated by UNG2 activity assays. Addition of PCNA (30–810 pmol) to standard uracil-DNA glycosylase reactions containing linear [uracil-3H]DNA stimulated UNG2 catalytic activity up to 2.6-fold. UNG2 activity was also stimulated by 7.5 mM MgCl2. The stimulatory effect of PCNA was increased by the addition of MgCl2; however, the dependence on PCNA concentration was the same, indicating that the effects of MgCl2 and PCNA on UNG2 activity occurred by independent mechanisms. Loading of PCNA onto the DNA substrate was required for stimulation, as the activity of UNG2 on circular DNA substrates was not affected by the addition of PCNA. Addition of replication factor C and ATP to reactions containing 90 pmol of PCNA resulted in two-fold stimulation of UNG2 activity on circular DNA. PMID:16216562

  3. Fish gotta swim, Birds gotta fly, I gotta do Feynmann Graphs 'til I die: A continuum Theory of Flocking

    NASA Astrophysics Data System (ADS)

    Toner, John; Tu, Yu-Hai

    2002-05-01

    We have developed a new continuum dynamical model for the collective motion of large "flocks" of biological organisms (e.g., flocks of birds, schools of fish, herds of wildebeest, hordes of bacteria, slime molds, etc.) . This model does for flocks what the Navier-Stokes equation does for fluids. The model predicts that, unlike simple fluids, flocks show huge fluctuation effects in spatial dimensions d < 4 that radically change their behavior. In d=2, it is only these effects that make it possible for the flock to move coherently at all. This explains why a million wildebeest can march together across the Serengeti plain, despite the fact that a million physicists gathered on the same plane could NOT all POINT in the same direction. Detailed quantitative predictions of this theory agree beautifully with computer simulations of flock motion.

  4. 75 FR 47662 - Self-Regulatory Organizations; The NASDAQ Stock Market LLC; Notice of Filing and Immediate...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-08-06

    ..., SNDK, SPY, T, TBT, TZA, UAUA, UNG, USO, UYG, V, VALE, VZ, WYNN, X, XHB, XLF, XRX and YHOO] * These fees..., SNDK, SPY, T, TBT, TZA, UAUA, UNG, USO, UYG, V, VALE, VZ, WYNN, X, XHB, XLF, XRX and YHOO. \\5\\...

  5. 75 FR 36134 - Self-Regulatory Organizations; International Securities Exchange, LLC; Notice of Filing and...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-06-24

    .... (``AIG''), American Express Company (``AXP''), Best Buy Company (``BBY''), Caterpillar, Inc. (``CAT..., IWM, XLF, AAPL, GE, JPM, INTC, GS, RIMM, T, VZ, UNG, FCX, CSCO, DIA, AMZN, X, AA, AIG, AXP, BBY, CAT..., IWM, XLF, AAPL, GE, JPM, INTC, GS, RIMM, T, VZ, UNG, FCX, CSCO, DIA, AMZN, X, AA, AIG, AXP, BBY,...

  6. Stream primary producers relate positively to watershed natural gas measures in north-central Arkansas streams.

    PubMed

    Austin, Bradley J; Hardgrave, Natalia; Inlander, Ethan; Gallipeau, Cory; Entrekin, Sally; Evans-White, Michelle A

    2015-10-01

    Construction of unconventional natural gas (UNG) infrastructure (e.g., well pads, pipelines) is an increasingly common anthropogenic stressor that increases potential sediment erosion. Increased sediment inputs into nearby streams may decrease autotrophic processes through burial and scour, or sediment bound nutrients could have a positive effect through alleviating potential nutrient limitations. Ten streams with varying catchment UNG well densities (0-3.6 wells/km(2)) were sampled during winter and spring of 2010 and 2011 to examine relationships between landscape scale disturbances associated with UNG activity and stream periphyton [chlorophyll a (Chl a)] and gross primary production (GPP). Local scale variables including light availability and water column physicochemical variables were measured for each study site. Correlation analyses examined the relationships of autotrophic processes and local scale variables with the landscape scale variables percent pasture land use and UNG metrics (well density and well pad inverse flow path length). Both GPP and Chl a were primarily positively associated with the UNG activity metrics during most sample periods; however, neither landscape variables nor response variables correlated well with local scale factors. These positive correlations do not confirm causation, but they do suggest that it is possible that UNG development can alleviate one or more limiting factors on autotrophic production within these streams. A secondary manipulative study was used to examine the link between nutrient limitation and algal growth across a gradient of streams impacted by natural gas activity. Nitrogen limitation was common among minimally impacted stream reaches and was alleviated in streams with high UNG activity. These data provide evidence that UNG may stimulate the primary production of Fayetteville shale streams via alleviation of N-limitation. Restricting UNG activities from the riparian zone along with better enforcement of

  7. Folate deficiency induces neurodegeneration and brain dysfunction in mice lacking uracil DNA glycosylase.

    PubMed

    Kronenberg, Golo; Harms, Christoph; Sobol, Robert W; Cardozo-Pelaez, Fernando; Linhart, Heinz; Winter, Benjamin; Balkaya, Mustafa; Gertz, Karen; Gay, Shanna B; Cox, David; Eckart, Sarah; Ahmadi, Michael; Juckel, Georg; Kempermann, Gerd; Hellweg, Rainer; Sohr, Reinhard; Hörtnagl, Heide; Wilson, Samuel H; Jaenisch, Rudolf; Endres, Matthias

    2008-07-09

    Folate deficiency and resultant increased homocysteine levels have been linked experimentally and epidemiologically with neurodegenerative conditions like stroke and dementia. Moreover, folate deficiency has been implicated in the pathogenesis of psychiatric disorders, most notably depression. We hypothesized that the pathogenic mechanisms include uracil misincorporation and, therefore, analyzed the effects of folate deficiency in mice lacking uracil DNA glycosylase (Ung-/-) versus wild-type controls. Folate depletion increased nuclear mutation rates in Ung-/- embryonic fibroblasts, and conferred death of cultured Ung-/- hippocampal neurons. Feeding animals a folate-deficient diet (FD) for 3 months induced degeneration of CA3 pyramidal neurons in Ung-/- but not Ung+/+ mice along with decreased hippocampal expression of brain-derived neurotrophic factor protein and decreased brain levels of antioxidant glutathione. Furthermore, FD induced cognitive deficits and mood alterations such as anxious and despair-like behaviors that were aggravated in Ung-/- mice. Independent of Ung genotype, FD increased plasma homocysteine levels, altered brain monoamine metabolism, and inhibited adult hippocampal neurogenesis. These results indicate that impaired uracil repair is involved in neurodegeneration and neuropsychiatric dysfunction induced by experimental folate deficiency.

  8. Human immunodeficiency virus type 1 Vpr protein binds to the uracil DNA glycosylase DNA repair enzyme.

    PubMed Central

    Bouhamdan, M; Benichou, S; Rey, F; Navarro, J M; Agostini, I; Spire, B; Camonis, J; Slupphaug, G; Vigne, R; Benarous, R; Sire, J

    1996-01-01

    The role of the accessory gene product Vpr during human immunodeficiency virus type 1 infection remains unclear. We have used the yeast two-hybrid system to identify cellular proteins that interact with Vpr and could be involved in its function. A cDNA clone which encodes the human uracil DNA glycosylase (UNG), a DNA repair enzyme involved in removal of uracil in DNA, has been isolated. Interaction between Vpr and UNG has been demonstrated by in vitro protein-protein binding assays using translated, radiolabeled Vpr and UNG recombinant proteins expressed as a glutathione S-transferase fusion protein. Conversely, purified UNG has been demonstrated to interact with Vpr recombinant protein expressed as a glutathione S-transferase fusion protein. Coimmunoprecipitation experiments confirmed that Vpr and UNG are associated within cells expressing Vpr. By using a panel of C- and N-terminally deleted Vpr mutants, we have determined that the core protein of Vpr, spanning amino acids 15 to 77, is involved in the interaction with UNG. We also demonstrate by in vitro experiments that the enzymatic activity of UNG is retained upon interaction with Vpr. PMID:8551605

  9. Breeding Value of the qSB9b and qSB12a QTLs in RiceBreeding Value of the qSB9b and qSB12a QTLs in Rice

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Sheath blight (SB) caused by Rhizoctonia solani Kuhn is a serious rice disease worldwide. The results of 123 TeQing-into-Lemont (TILs) showed those with introgressions containing qSB9b and/or qSB12a were among the most SB resistant TILs. TIL:615, TIL:642 and TIL:567 have consistently appeared modera...

  10. 77 FR 46776 - Self-Regulatory Organizations; International Securities Exchange, LLC; Notice of Filing and...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-08-06

    ... Symbols'' are options overlying QQQ, C, BAC, SPY, IWM, XLF, GE, JPM, INTC, RIMM, T, VZ, UNG, FCX, CSCO... efficiently, please use only one method. The Commission will post all comments on the Commission's...

  11. 75 FR 56627 - Self-Regulatory Organizations; Chicago Board Options Exchange, Incorporated; Notice of Filing and...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-09-16

    ...: BAC, C, DXD, EMC, EWJ, F, FAX, FAZ, GE, INTC, MOT, MSFT, MU, NOK, Q, QID, S, SIRI, SKF, T, TWM, UNG... and review your comments more efficiently, please use only one method. The Commission will post...

  12. 75 FR 61549 - Self-Regulatory Organizations; Chicago Board Options Exchange, Incorporated; Notice of Filing and...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-10-05

    ...: BAC, C, DXD, EMC, EWJ, F, FAX, FAZ, GE, INTC, MOT, MSFT, MU, NOK, Q, QID, S, SIRI, SKF, T, TWM, UNG... and review your comments more efficiently, please use only one method. The Commission will post...

  13. 76 FR 17462 - Self-Regulatory Organizations; International Securities Exchange, LLC; Notice of Filing and...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-03-29

    ... in these select symbols: QQQQ, C, BAC, SPY, IWM, XLF, GE, JPM, INTC, RIMM, T, VZ, UNG, FCX, CSCO, DIA... efficiently, please use only one method. The Commission will post all comments on the Commission's...

  14. Elevated levels of plasma homocysteine, deficiencies in dietary folic acid and uracil-DNA glycosylase impair learning in a mouse model of vascular cognitive impairment.

    PubMed

    Jadavji, Nafisa M; Farr, Tracy D; Lips, Janet; Khalil, Ahmed A; Boehm-Sturm, Philipp; Foddis, Marco; Harms, Christoph; Füchtemeier, Martina; Dirnagl, Ulrich

    2015-04-15

    Dietary deficiencies in folic acid result in elevated levels of plasma homocysteine, which has been associated with the development of dementia and other neurodegenerative disorders. Previously, we have shown that elevated levels of plasma homocysteine in mice deficient for a DNA repair enzyme, uracil-DNA glycosylase (UNG), result in neurodegeneration. The goal of this study was to evaluate how deficiencies in folic acid and UNG along with elevated levels of homocysteine affect vascular cognitive impairment, via chronic hypoperfursion in an animal model. Ung(+/+) and Ung(-/-) mice were placed on either control (CD) or folic acid deficient (FADD) diets. Six weeks later, the mice either underwent implantation of microcoils around both common carotid arteries. Post-operatively, behavioral tests began at 3-weeks, angiography was measured after 5-weeks using MRI to assess vasculature and at completion of study plasma and brain tissue was collected for analysis. Learning impairments in the Morris water maze (MWM) were observed only in hypoperfused Ung(-/-) FADD mice and these mice had significantly higher plasma homocysteine concentrations. Interestingly, Ung(+/+) FADD produced significant remodeling of the basilar artery and arterial vasculature. Increased expression of GFAP was observed in the dentate gyrus of Ung(-/-) hypoperfused and FADD sham mice. Chronic hypoperfusion resulted in increased cortical MMP-9 protein levels of FADD hypoperfused mice regardless of genotypes. These results suggest that elevated levels of homocysteine only, as a result of dietary folic acid deficiency, don't lead to memory impairments and neurobiochemical changes. Rather a combination of either chronic hypoperfusion or UNG deficiency is required.

  15. 'Faking til you make it': social capital accumulation of individuals on low incomes living in contrasting socio-economic neighbourhoods and its implications for health and wellbeing.

    PubMed

    Browne-Yung, Kathryn; Ziersch, Anna; Baum, Fran

    2013-05-01

    People on low-income living in low socio-economic neighbourhoods have poorer health in comparison with those living in advantaged neighbourhoods. To explore neighbourhood effects on health and social capital creation, the experiences of low-income people living in contrasting socio-economic neighbourhoods were compared, in order to examine how low-income status and differing levels of neighbourhood resources contributed to perceived health and wellbeing. Quantitative and qualitative data were analysed: survey data from 601 individuals living in contrasting socio-economic areas and in-depth interviews with a new sample of 24 individuals on low-incomes. The study was guided by Bourdieu's theory of practice, which examines how social inequalities are created and reproduced through the relationship between individuals' varying resources of economic, social and cultural capital. This included an examination of individual life histories, cultural distinction and how social positions are reproduced. Participants' accounts of their early life experience showed how parental socio-economic position and socially patterned events taking place across the life course, created different opportunities for social network creation, choice of neighbourhood and levels of resources available throughout life, all of which can influence health and wellbeing. A definition of poverty by whether an individual or household has sufficient income at a particular point in time was an inadequate measure of disadvantage. This static measure of 'low income' as a category disguised a number of different ways in which disadvantage was experienced or, conversely, how life course events could mitigate the impact of low-income. This study found that the resources necessary to create social capital such as cultural capital and the ability to socially network, differed according to the socio-economic status of the neighbourhood, and that living in an advantaged area does not automatically guarantee access to potentially beneficial social networks.

  16. Don't Know What You've Got 'Til It's Gone? Skills-Led Qualifications, Secondary School Attainment and Policy Choices

    ERIC Educational Resources Information Center

    Harrison, Neil; James, David; Last, Kathryn

    2015-01-01

    In the name of curriculum breadth and raising standards, recent government policy in England has removed a large number of non-academic qualifications from the list of those that secondary schools can count in league tables, discouraging their use. Most of these were vocational qualifications, but they also include skills-led qualifications. This…

  17. Impact of Marcellus Shale natural gas development in southwest Pennsylvania on volatile organic compound emissions and regional air quality.

    PubMed

    Swarthout, Robert F; Russo, Rachel S; Zhou, Yong; Miller, Brandon M; Mitchell, Brittney; Horsman, Emily; Lipsky, Eric; McCabe, David C; Baum, Ellen; Sive, Barkley C

    2015-03-03

    The Marcellus Shale is the largest natural gas deposit in the U.S. and rapid development of this resource has raised concerns about regional air pollution. A field campaign was conducted in the southwestern Pennsylvania region of the Marcellus Shale to investigate the impact of unconventional natural gas (UNG) production operations on regional air quality. Whole air samples were collected throughout an 8050 km(2) grid surrounding Pittsburgh and analyzed for methane, carbon dioxide, and C1-C10 volatile organic compounds (VOCs). Elevated mixing ratios of methane and C2-C8 alkanes were observed in areas with the highest density of UNG wells. Source apportionment was used to identify characteristic emission ratios for UNG sources, and results indicated that UNG emissions were responsible for the majority of mixing ratios of C2-C8 alkanes, but accounted for a small proportion of alkene and aromatic compounds. The VOC emissions from UNG operations accounted for 17 ± 19% of the regional kinetic hydroxyl radical reactivity of nonbiogenic VOCs suggesting that natural gas emissions may affect compliance with federal ozone standards. A first approximation of methane emissions from the study area of 10.0 ± 5.2 kg s(-1) provides a baseline for determining the efficacy of regulatory emission control efforts.

  18. Evaluation of the prognostic value of tumor-infiltrating lymphocytes in triple-negative breast cancers

    PubMed Central

    Tian, Tian; Ruan, Miao; Yang, Wentao; Shui, Ruohong

    2016-01-01

    Tumor-infiltrating lymphocytes (TILs) may be associated with clinical outcome in triple-negative breast cancers (TNBCs). However, lacking of standardized methodologies in TILs evaluation has hindered its application in clinical practice. To evaluate the prognostic role of TILs scored by methods recommended by International TILs Working Group 2014, we performed a retrospective study of TILs in 425 primary invasive TNBCs in a Chinese population with a median follow-up of 4 years. Intratumoral TILs (iTILs) and stromal TILs (sTILs) were scored respectively. The associations between TILs and disease-free survival (DFS), distant disease-free survival (DDFS) and overall survival (OS) were evaluated with COX models. ITILs were not associated with prognosis. Higher sTILs were associated with better prognosis; for every 10% increase in sTILs, a 5% reduction of risk of recurrence or death (P < 0.001), 5% reduction of risk of distant recurrence (P < 0.001), and 4% reduction of risk of death (P = 0.002) were observed. Multivariate analysis confirmed sTILs to be an independent prognostic marker. 3.5% of TNBCs had more than 50% lymphocytes (lymphocyte-predominant breast cancer, LPBC), and associations between LPBC status and prognosis were observed but did not reach statistical significance. TNBCs with more than 20% sTILs had a significantly better prognosis than the patients with no more than 20% sTILs. In conclusion, our study indicated that sTILs scored by methods recommended by International TILs Working Group 2014 were associated with the prognosis of TNBCs. STILs could be an independent prognostic biomarker in TNBCs, increasing sTILs predicting better prognosis. PMID:27323808

  19. Uracil-DNA glycosylase-treated reverse transcription loop-mediated isothermal amplification for rapid detection of avian influenza virus preventing carry-over contamination

    PubMed Central

    Kim, Eun-Mi; Jeon, Hyo-Sung; Kim, Ji-Jung; Shin, Yeun-Kyung; Lee, Youn-Jeong; Yeo, Sang-Geon

    2016-01-01

    Here, we describe a uracil-DNA glycosylase (UNG)-treated reverse transcription loop-mediated isothermal amplification (uRT-LAMP) for the visual detection of all subtypes of avian influenza A virus (AIV). The uRT-LAMP assay can prevent unwanted amplification by carryover contamination of the previously amplified DNA, although the detection limit of the uRT-LAMP assay is 10-fold lower than that of the RT-LAMP without a UNG treatment. To the best of our knowledge, this is the first successful application of deoxyuridine triphosphate/UNG strategy in RT-LAMP for AIV detection, and the assay can be applied for the rapid, and reliable diagnosis of AIVs, even in contaminated samples. PMID:26726027

  20. Uracil-DNA glycosylase-treated reverse transcription loop-mediated isothermal amplification for rapid detection of avian influenza virus preventing carry-over contamination.

    PubMed

    Kim, Eun-Mi; Jeon, Hyo-Sung; Kim, Ji-Jung; Shin, Yeun-Kyung; Lee, Youn-Jeong; Yeo, Sang-Geon; Park, Choi-Kyu

    2016-09-30

    Here, we describe a uracil-DNA glycosylase (UNG)-treated reverse transcription loop-mediated isothermal amplification (uRT-LAMP) for the visual detection of all subtypes of avian influenza A virus (AIV). The uRT-LAMP assay can prevent unwanted amplification by carryover contamination of the previously amplified DNA, although the detection limit of the uRT-LAMP assay is 10-fold lower than that of the RT-LAMP without a UNG treatment. To the best of our knowledge, this is the first successful application of deoxyuridine triphosphate/UNG strategy in RT-LAMP for AIV detection, and the assay can be applied for the rapid, and reliable diagnosis of AIVs, even in contaminated samples.

  1. Increased uracil insertion in DNA is cytotoxic and increases the frequency of mutation, double strand break formation and VSG switching in Trypanosoma brucei.

    PubMed

    Castillo-Acosta, Víctor M; Aguilar-Pereyra, Fernando; Bart, Jean-Mathieu; Navarro, Miguel; Ruiz-Pérez, Luis M; Vidal, Antonio E; González-Pacanowska, Dolores

    2012-12-01

    Deoxyuridine 5'-triphosphate pyrophosphatase (dUTPase) and uracil-DNA glycosylase (UNG) are key enzymes involved in the control of the presence of uracil in DNA. While dUTPase prevents uracil misincorporation by removing dUTP from the deoxynucleotide pool, UNG excises uracil from DNA as a first step of the base excision repair pathway (BER). Here, we report that strong down-regulation of dUTPase in UNG-deficient Trypanosoma brucei cells greatly impairs cell viability in both bloodstream and procyclic forms, underscoring the extreme sensitivity of trypanosomes to uracil in DNA. Depletion of dUTPase activity in the absence of UNG provoked cell cycle alterations, massive dUTP misincorporation into DNA and chromosomal fragmentation. Overall, trypanosomatid cells that lack dUTPase and UNG activities exhibited greater proliferation defects and DNA damage than cells deficient in only one of these activities. To determine the mutagenic consequences of uracil in DNA, mutation rates and spectra were analyzed in dUTPase-depleted cells in the presence of UNG activity. These cells displayed a spontaneous mutation rate 9-fold higher than the parental cell line. Base substitutions at A:T base pairs and deletion frequencies were both significantly enhanced which is consistent with the generation of mutagenic AP sites and DNA strand breaks. The increase in strand breaks conveyed a concomitant increase in VSG switching in vitro. The low tolerance of T. brucei to uracil in DNA emphasizes the importance of uracil removal and regulation of intracellular dUTP pool levels in cell viability and genetic stability and suggests potential strategies to compromise parasite survival.

  2. Human gene transfer: Characterization of human tumor-infiltrating lymphocytes as vehicles for retroviral-mediated gene transfer in man

    SciTech Connect

    Kasid, A.; Morecki, S.; Aebersold, P.; Cornetta, K.; Culver, K.; Freeman, S.; Director, E.; Lotze, M.T.; Blaese, R.M.; Anderson, W.F.; Rosenberg, S.A. )

    1990-01-01

    Tumor-infiltrating lymphocytes (TILs) are cells generated from tumor suspensions cultured in interleukin 2 that can mediate cancer regression when adoptively transferred into mice or humans. Since TILs proliferate rapidly in vitro, recirculate, and preferentially localize at the tumor site in vivo, they provide an attractive model for delivery of exogenous genetic material into man. To determine whether efficient gene transfer into TILs is feasible. The authors transduced human TILs with the bacterial gene for neomycin-resistance (Neo{sup R}) using the retroviral vector N2. The transduced TIL populations were stable and polyclonal with respect to the intact Neo{sup R} gene integration and expressed high levels of neomycin phosphotransferase activity. The Neo{sup R} gene insertion did not alter the in vitro growth pattern and interleukin 2 dependence of the transduced TILs. Analyses of T-cell receptor gene rearrangement for {beta}- and {gamma}-chain genes revealed the oligoclonal nature of the TIL populations with no major change in the DNA rearrangement patterns or the levels of mRNA expression of the {beta} and {gamma} chains following transduction and selection of TILs in the neomycin analog G418. Human TILs expressed mRNA for tumor necrosis factors ({alpha} and {beta}) and interleukin 2 receptor P55. This pattern of cytokine-mRNA expression was not significantly altered following the transduction of TILs. The studies demonstrate the feasibility of TILs as suitable cellular vehicles for the introduction of therapeutic genes into patients receiving autologous TILs.

  3. Tumour infiltrating lymphocytes correlate with improved survival in patients with esophageal squamous cell carcinoma

    PubMed Central

    Jiang, Dongxian; Liu, Yalan; Wang, Hao; Wang, Haixing; Song, Qi; Sujie, Akesu; Huang, Jie; Xu, Yifan; Zeng, Haiying; Tan, Lijie; Hou, Yingyong; Xu, Chen

    2017-01-01

    We undertook a study of tumour infiltrating lymphocytes (TILs) in a large and relatively homogeneous group of patients with completely resected esophageal squamous cell carcinoma (ESCC). Hematoxylin and eosin–stained sections of 235 ESCC tumours were evaluated for density of TILs in intratumoural (iTIL) and stromal compartments (sTIL). Foxp3+, CD4+, and CD8+ T cells in tumoural and stromal areas were evaluated by immunohistochemistry. Of the 235 tumours, high sTIL (>10%), and iTIL (>10%) were observed in 101 (43.0%) and 98 (41.7%), respectively. The median follow-up period was 36.0 months (95% CI 29.929–42.071). Univariate analysis revealed that sTIL (>10%), iTIL (>20%), vessels involvement, lymph node metastasis, and clinical stage were significantly associated with postoperative outcome. In multivariate analysis, high sTIL (HR: 0.664, P = 0.019 for Disease free survival; HR: 0.608, P = 0.005 for Overall survival) was identified as independent better prognostic factor. Further analysis, sTIL was identified as independently prognostic factor in Stage III-IVa disease, which was not found in Stage I-II disease. Our study demonstrated that sTIL was associated with better ESCC patients’ survival, especially in Stage III-IVa disease. Assessment of sTIL could be useful to discriminate biological behavior for ESCC patients. PMID:28322245

  4. Tumour infiltrating lymphocytes correlate with improved survival in patients with esophageal squamous cell carcinoma.

    PubMed

    Jiang, Dongxian; Liu, Yalan; Wang, Hao; Wang, Haixing; Song, Qi; Sujie, Akesu; Huang, Jie; Xu, Yifan; Zeng, Haiying; Tan, Lijie; Hou, Yingyong; Xu, Chen

    2017-03-21

    We undertook a study of tumour infiltrating lymphocytes (TILs) in a large and relatively homogeneous group of patients with completely resected esophageal squamous cell carcinoma (ESCC). Hematoxylin and eosin-stained sections of 235 ESCC tumours were evaluated for density of TILs in intratumoural (iTIL) and stromal compartments (sTIL). Foxp3+, CD4+, and CD8+ T cells in tumoural and stromal areas were evaluated by immunohistochemistry. Of the 235 tumours, high sTIL (>10%), and iTIL (>10%) were observed in 101 (43.0%) and 98 (41.7%), respectively. The median follow-up period was 36.0 months (95% CI 29.929-42.071). Univariate analysis revealed that sTIL (>10%), iTIL (>20%), vessels involvement, lymph node metastasis, and clinical stage were significantly associated with postoperative outcome. In multivariate analysis, high sTIL (HR: 0.664, P = 0.019 for Disease free survival; HR: 0.608, P = 0.005 for Overall survival) was identified as independent better prognostic factor. Further analysis, sTIL was identified as independently prognostic factor in Stage III-IVa disease, which was not found in Stage I-II disease. Our study demonstrated that sTIL was associated with better ESCC patients' survival, especially in Stage III-IVa disease. Assessment of sTIL could be useful to discriminate biological behavior for ESCC patients.

  5. Increased CD4 and CD8-positive T cell infiltrate signifies good prognosis in a subset of triple-negative breast cancer.

    PubMed

    Matsumoto, Hirofumi; Thike, Aye Aye; Li, Huihua; Yeong, Joe; Koo, Si-Lin; Dent, Rebecca Alexandra; Tan, Puay Hoon; Iqbal, Jabed

    2016-04-01

    Tumour-infiltrating lymphocytes (TILs) signify immune response to tumour in a variety of cancers including breast cancer. However, earlier studies examining the clinical significance of TILs in breast cancers have generated mixed results. There are only a few that address the relationship between TILs and clinical outcomes in triple-negative breast cancers (TNBC). The aim of this study is to evaluate the clinical significance of TILs that express CD4 + and CD8 + , in TNBC. Immunohistochemical staining of CD4 and CD8 was performed on tissue microarrays of 164 cases of TNBC. TILs were counted separately as intratumoral when within the cancer cell nests (iTILs) and as stromal when within cancer stroma (sTILs). High CD8 + iTILs and sTILs, and CD4 + iTILs correlated with histologic grade. On Kaplan-Meier analysis, a significantly better survival rate was observed in high CD8 + iTIL (disease-free survival, DFS: P = 0.004, overall survival, OS: P = 0.02) and both high CD4 + iTILs (DFS: P = 0.025, OS: P = 0.023) and sTILs (DFS: P = 0.01, OS: P = 0.002). In multivariate analysis, CD8 + iTILs (DFS: P = 0.0095), CD4 + sTILs (DFS: P = 0.0084; OS: P = 0.0118), and CD4 (high) CD8 (high) CD8 iTILs (DFS: P = 0.0121; OS: P = 0.0329) and sTILs (DFS: P = 0.0295) showed significantly better survival outcomes. These results suggest that high levels of both CD8 + iTILs and CD4 + sTILs as well as CD4 (high) CD8 (high) iTILs and sTILs are independent prognostic factors in TNBC.

  6. Prognostic value of tumor-infiltrating lymphocytes on residual disease after primary chemotherapy for triple-negative breast cancer: a retrospective multicenter study

    PubMed Central

    Dieci, M. V.; Criscitiello, C.; Goubar, A.; Viale, G.; Conte, P.; Guarneri, V.; Ficarra, G.; Mathieu, M. C.; Delaloge, S.; Curigliano, G.; Andre, F.

    2014-01-01

    Background There is a need to develop surrogates for treatment efficacy in the neoadjuvant setting to speed-up drug development and stratify patients according to outcome. Preclinical studies showed that chemotherapy induces an antitumor immune response. In order to develop new surrogates for drug efficacy, we assessed the prognostic value of tumor-infiltrating lymphocytes (TIL) on residual disease after neoadjuvant chemotherapy (NACT) in patients with triple-negative breast cancer (TNBC). Patients and methods Three hundred four TNBC patients with residual disease after NACT were retrospectively identified in three different hospitals. Hematoxylin and eosin-stained slides from surgical postchemotherapy specimens were evaluated for intratumoral (It-TIL) and stromal (Str-TIL) TIL. Cases were classified as High-TIL if It-TIL and/or Str-TIL >60%. Results TIL were assessable for 278 cases. Continuous It-TIL and Str-TIL variables were strong prognostic factors in the multivariate model, both for metastasis-free [hazard ratio (HR) 0.86, 95% confidence interval (CI) 0.77–0.96, P = 0.01 and HR 0.85, 95% CI 0.75–0.98, P = 0.02 for Str-TIL and It-TIL, respectively] and overall survival (HR 0.86, 95% CI 0.77–0.97, P = 0.01 and HR 0.86, 95% CI 0.75–0.99, P = 0.03 for Str-TIL and It-TIL, respectively). The 5-year overall survival rate was 91% (95% CI 68% to 97%) for High-TIL patients (n = 27) and 55% (95% CI 48% to 61%) for Low-TIL patients (HR 0.19, 95% CI 0.06–0.61, log-rank P = 0.0017). The major prognostic impact of TIL was seen for patients with large tumor burden following NACT (residual tumor >2 cm and/or node metastasis). In all but one High-TIL case, It-TIL and Str-TIL values were lower on the prechemotherapy sample. Conclusions The presence of TIL in residual disease after NACT is associated with better prognosis in TNBC patients. This parameter may represent a new surrogate of drug efficacy to test investigational agents in the neoadjuvant setting and a new

  7. Integrating the UAS in Undergraduate Teaching and Research - Opportunities and Challenges at University of North Georgia

    NASA Astrophysics Data System (ADS)

    Sharma, J. B.; Hulsey, D.

    2014-11-01

    The University of North Georgia (UNG) has begun to evaluate both fixed and rotary UAS platforms across the departments to evaluate their potential for furthering both student learning experiences and undergraduate research. A research project of the Institute for Spatial Analysis (IESA) at UNG has acquired the fixed wing eBee UAS and is currently piloting its integration into the undergraduate geospatial science curriculum. Limited very low altitude, line of sight calibration runs within areas of our campus have help us understand the capabilities that this technology brings to learning and research opportunities at UNG. In our pilot area of study on the UNG Gainesville Campus, we will collect overlapping imagery and generate 3-D models. These models will be compared with models based on LiDAR data. Geographic Object Based Image Analysis (GEOBIA) methods are essential to the analysis of voluminous high resolution UAS data and the associated computational and regulatory issues are discussed. Several future interdisciplinary projects are envisaged with the eBee UAS and this project helps establish their viability.

  8. Band Model Calculations of Atmospheric Transmittance for Hot Gas Line Emission Sources

    DTIC Science & Technology

    1974-12-31

    influenced by the source spectral characteristics. The present work is concerned with the design and applicacion of a band modr.Ung technique that...optics, chemical reactions in polluted atmospheres, chemical reactiona of excited species in rocket plumes, chemical thermodynamics , plasma and

  9. 75 FR 20413 - Self-Regulatory Organizations; NYSE Arca, Inc.; Notice of Filing and Immediate Effectiveness of a...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-04-19

    .... Electronic executions in options overlying SPY, C, BAC, QQQQ, AAPL, IWM, XLF, GLD, EEM, GE, UNG, FAZ, DIA... credit above the stated Post Liquidity credit. This is consistent with similar billing treatment of... Market Maker Post Liquidity Incentive Credit that provided Market Makers with an additional $0.01...

  10. 76 FR 18274 - Self-Regulatory Organizations; Notice of Filing and Immediate Effectiveness of Proposed Rule...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-04-01

    ... credit above the stated post liquidity credit for electronic transactions in options overlying SPY, C, BAC, QQQQ, AAPL, IWM, XLF, GLD, EEM, GE, UNG, FAZ, DIA, GDX, and USO which are referred to as the... one method. The Commission will post all comments on the Commission's Internet Web site (...

  11. Relevance of underground natural gas storage to geologic sequestration of carbon dioxide

    SciTech Connect

    Lippmann, Marcelo J.; Benson, Sally M.

    2002-07-01

    The practice of underground natural gas storage (UNGS), which started in the USA in 1916, provides useful insight into the geologic sequestration of carbon dioxide--the dominant anthropogenic greenhouse gas released into the atmosphere. In many ways, UNGS is directly relevant to geologic CO{sub 2} storage because, like CO{sub 2}, natural gas (essentially methane) is less dense than water. Consequently, it will tend to rise to the top of any subsurface storage structure located below the groundwater table. By the end of 2001 in the USA, about 142 million metric tons of natural gas were stored underground in depleted oil and gas reservoirs and brine aquifers. Based on their performance, UNGS projects have shown that there is a safe and effective way of storing large volumes of gases in the subsurface. In the small number of cases where failures did occur (i.e., leakage of the stored gas into neighboring permeable layers), they were mainly related to improper well design, construction, maintenance, and/or incorrect project operation. In spite of differences in the chemical and physical properties of the gases, the risk-assessment, risk-management, and risk-mitigation issues relevant to UNGS projects are also pertinent to geologic CO{sub 2} sequestration.

  12. Probabilistic and Stochastic Methods in Analysis, with Applications

    DTIC Science & Technology

    1992-01-01

    condizione di chiusura di un sistema di funzioni ortogo- nali. Atti R. Accad. Naz. Lincei, Rend. Cl. Sci. Fis. Mat. Nat, 30:498-501, 1921. [52] J. von Neumann...Zimmer and V. Neuhoff, editors, Kolloquiun) DFG-Schwerpunktprogramni Digitale Signalverarbeit ung, pages 1-4, Gdttingen, 1981. [22] R.G. Wiley. Recovery

  13. Telling Their Stories: Women Construct/Instruct through Survival Rhetoric.

    ERIC Educational Resources Information Center

    Meagher, Eileen M.

    Malika Oufkir of Morocco recounts her story in "Stolen Lives." Loung Ung of Cambodia relates her story in "First, They Killed My Father." Susan McDougal of Arkansas, USA, tells her story in the aptly named, "The Woman Who Wouldn't Talk." This paper looks at the struggles of these three very different women from very…

  14. Single nucleotide polymorphisms in uracil-processing genes, intake of one-carbon nutrients and breast cancer risk

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Background/Objectives: The misincorporation of uracil into DNA leads to genomic instability. In a previous study, some of us identified four common single nucleotide polymorphisms (SNPs) in uracil-processing genes (rs2029166 and rs7296239 in SMUG1, rs34259 in UNG and rs4775748 in DUT) that were asso...

  15. Mechanism of mutation by thymine starvation in Escherichia coli: clues from mutagenic specificity.

    PubMed Central

    Kunz, B A; Glickman, B W

    1985-01-01

    To probe the mechanisms of mutagenesis induced by thymine starvation, we examined the mutational specificity of this treatment in strains of Escherichia coli that are wild type (Ung+) or deficient in uracil-DNA-glycosylase (Ung-). An analysis of Ung+ his-4 (ochre) revertants revealed that the majority of induced DNA base substitution events were A:T----G:C transitions. However, characterization of lacI nonsense mutations induced by thymine starvation demonstrated that G:C----A:T transitions and all four possible transversions also occurred. In addition, thymineless episodes led to reversion of the trpE9777 frameshift allele. Although the defect in uracil-DNA-glycosylase did not appear to affect the frequency of total mutations induced in lacI by thymine deprivation, the frequency of nonsense mutations was reduced by 30%, and the spectrum of nonsense mutations was altered. Furthermore, the reversion of trpE9777 was decreased by 90% in the Ung- strain. These findings demonstrate that in E. coli, thymine starvation can induce frameshift mutations and all types of base substitutions. The analysis of mutational specificity indicates that more than a single mechanism is involved in the induction of mutation by thymine depletion. We suggest that deoxyribonucleoside triphosphate pool imbalances, the removal of uracil incorporated into DNA during thymine starvation, and the induction of recA-dependent DNA repair functions all may play a role in thymineless mutagenesis. PMID:3888966

  16. Social Influence, Health Variables and Criminal Behaviours Associated with Substance Use among Rural Norwegian Adolescents

    ERIC Educational Resources Information Center

    Nordfjaern, Trond; Dahl, Hilde; Flemmen, Grete

    2013-01-01

    Aims: To investigate social influence, health, criminality and substance use in a sample of 1288 Norwegian rural adolescents. Relations between these factors and substance use were examined. Methods: Data were obtained from the "UngData" study. A cross-sectional questionnaire survey was conducted among adolescents (n = 740) in nine…

  17. ANALYSIS OF UNCERTAINTIES IN DOSE RECONSTRUCTION FROM BIOMARKERS: IMPACT ON STUDY DESIGN

    EPA Science Inventory

    The absorbed dose is defined as the quantity which has passed through the barriers (skin, GI tract, The absorbed dose of a pesticide can be estimated from its established urinary biomarker. ungs). For an exposure study, there are several options for biomarker collection, each w...

  18. Tumor-Infiltrating Lymphocyte Grade in Primary Melanomas Is Independently Associated With Melanoma-Specific Survival in the Population-Based Genes, Environment and Melanoma Study

    PubMed Central

    Thomas, Nancy E.; Busam, Klaus J.; From, Lynn; Kricker, Anne; Armstrong, Bruce K.; Anton-Culver, Hoda; Gruber, Stephen B.; Gallagher, Richard P.; Zanetti, Roberto; Rosso, Stefano; Dwyer, Terence; Venn, Alison; Kanetsky, Peter A.; Groben, Pamela A.; Hao, Honglin; Orlow, Irene; Reiner, Anne S.; Luo, Li; Paine, Susan; Ollila, David W.; Wilcox, Homer; Begg, Colin B.; Berwick, Marianne

    2013-01-01

    Purpose Although most hospital-based studies suggest more favorable survival with tumor-infiltrating lymphocytes (TILs) present in primary melanomas, it is uncertain whether TILs provide prognostic information beyond existing melanoma staging definitions. We addressed the issue in an international population-based study of patients with single and multiple primary melanomas. Patients and Methods On the basis of the Genes, Environment and Melanoma (GEM) study, we conducted follow-up of 2,845 patients diagnosed from 1998 to 2003 with 3,330 invasive primary melanomas centrally reviewed for TIL grade (absent, nonbrisk, or brisk). The odds of TIL grades associated with clinicopathologic features and survival by TIL grade were examined. Results Independent predictors (P < .05) for nonbrisk TIL grade were site, histologic subtype, and Breslow thickness, and for brisk TIL grade, they were age, site, Breslow thickness, and radial growth phase. Nonbrisk and brisk TIL grades were each associated with lower American Joint Committee on Cancer (AJCC) tumor stage compared with TIL absence (Ptrend < .001). Death as a result of melanoma was 30% less with nonbrisk TIL grade (hazard ratio [HR], 0.7; 95% CI, 0.5 to 1.0) and 50% less with brisk TIL grade (HR, 0.5; 95% CI, 0.3 to 0.9) relative to TIL absence, adjusted for age, sex, site, and AJCC tumor stage. Conclusion At the population level, higher TIL grade of primary melanoma is associated with a lower risk of death as a result of melanoma independently of tumor characteristics currently used for AJCC tumor stage. We conclude that TIL grade deserves further prospective investigation to determine whether it should be included in future AJCC staging revisions. PMID:24127443

  19. Tumor infiltrating lymphocytes in ovarian cancer.

    PubMed

    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.

  20. The emergence of non-cytolytic NK1.1+ T cells in the long-term culture of murine tumour-infiltrating lymphocytes: a possible role of transforming growth factor-beta.

    PubMed Central

    Tamada, K; Harada, M; Ito, O; Takenoyama, M; Mori, T; Matsuzaki, G; Nomoto, K

    1996-01-01

    The mechanism by which murine tumour-infiltrating lymphocytes (TIL) decreased their anti-tumour activity during an in vitro culture with interleukin-2 (IL-2) was investigated. A phenotype analysis revealed that the TIL cultured for 7 days (TIL-d7) were exclusively NKI.1- CD4- CD8+ CD3+ cells and that this population was replaced by natural killer (NK)1.1+ CD4- CD8 CD3+ cells by day 27 (TIL-d27) during the culture of TIL. The TIL-d7 cells showed a cytolytic activity against B16 melanoma, whereas the TIL-d27 cells had lost this activity, suggesting that the decrease in the anti tumour effect of TIL during the culture with IL-2 was due to their populational change. Analysis on the characteristics of the TIL-d27 cells revealed that they expressed skewed T-cell receptor (TCR) V beta 5 and increased mRNA expression of V alpha 14. In addition, they expressed transforming growth factor beta (TGF-beta) mRNA. Interestingly, TGF-beta augmented the proliferation of TIL-d27 cells under the presence of IL-2, but suppressed that of TIL-d7 cells. Moreover, the proliferation of TIL-d27 cells was suppressed by anti-TGF-beta monoclonal antibody. Collectively, these results suggest that, in contrast to its suppressive effect on anti-tumour effector T cells. TGF-beta could be an autocrine growth factor for NKL1.1+ T cells and thereby induce non-cytolytic NK1.1+ T cells in the long-term culture of TIL. Images Figure 4 Figure 6 PMID:9014832

  1. Similar lymphocytic infiltration pattern in primary breast cancer and their corresponding distant metastases.

    PubMed

    Sobottka, Bettina; Pestalozzi, Bernhard; Fink, Daniel; Moch, Holger; Varga, Zsuzsanna

    2016-06-01

    Tumor infiltrating lymphocytes in primary breast cancer (TIL) are acknowledged measures of disease free survival (DFS) in adjuvant and neoadjuvant settings. Little is known about the biology of metastasis infiltrating lymphocytes (mTIL) although the local immunity of the metastatic site may critically influence the infiltrate composite. To address this question, we compared mTIL with their matched TIL in 87 breast cancer patients and their corresponding distant metastasis at four different anatomical locations. Sections of surgical specimen were immunohistochemically analyzed for CD4(+), CD8(+) and CD20(+) lymphocytes in three different tumor compartments: intratumoral lymphocytes (iTIL) defined as lymphocytes in direct contact with breast cancer cells, stromal lymphocytes (sTIL) located within the intratumoral stromal tissue and invasive-margin lymphocytes (imTIL). Overall, we found fewer (p < 0.001) mTIL than TIL. Within the tumor compartments, imTIL were more frequent than sTIL and iTIL both within metastases and the matched primary tumors (PT) (p < 0.001). CD4(+) T cells were more numerous than CD8(+) T cells and CD20(+) B cells (p < 0.001). There was a similar pattern in PT and their corresponding metastasis. Only patients with brain metastases differed from the others displaying less CD20(+) B cells at the infiltrative margin of the PT (p < 0.05). In summary, mTIL were significantly reduced within metastases but still mirrored the infiltrate pattern of the PT, interestingly regardless of the metastatic anatomical locations investigated. Our results suggest that the PT assigns the infiltrating lymphocyte pattern resumed at the metastatic site.

  2. Similar lymphocytic infiltration pattern in primary breast cancer and their corresponding distant metastases

    PubMed Central

    Sobottka, Bettina; Pestalozzi, Bernhard; Fink, Daniel; Moch, Holger; Varga, Zsuzsanna

    2016-01-01

    ABSTRACT Tumor infiltrating lymphocytes in primary breast cancer (TIL) are acknowledged measures of disease free survival (DFS) in adjuvant and neoadjuvant settings. Little is known about the biology of metastasis infiltrating lymphocytes (mTIL) although the local immunity of the metastatic site may critically influence the infiltrate composite. To address this question, we compared mTIL with their matched TIL in 87 breast cancer patients and their corresponding distant metastasis at four different anatomical locations. Sections of surgical specimen were immunohistochemically analyzed for CD4+, CD8+ and CD20+ lymphocytes in three different tumor compartments: intratumoral lymphocytes (iTIL) defined as lymphocytes in direct contact with breast cancer cells, stromal lymphocytes (sTIL) located within the intratumoral stromal tissue and invasive-margin lymphocytes (imTIL). Overall, we found fewer (p < 0.001) mTIL than TIL. Within the tumor compartments, imTIL were more frequent than sTIL and iTIL both within metastases and the matched primary tumors (PT) (p < 0.001). CD4+ T cells were more numerous than CD8+ T cells and CD20+ B cells (p < 0.001). There was a similar pattern in PT and their corresponding metastasis. Only patients with brain metastases differed from the others displaying less CD20+ B cells at the infiltrative margin of the PT (p < 0.05). In summary, mTIL were significantly reduced within metastases but still mirrored the infiltrate pattern of the PT, interestingly regardless of the metastatic anatomical locations investigated. Our results suggest that the PT assigns the infiltrating lymphocyte pattern resumed at the metastatic site. PMID:27471624

  3. Reliability and Validity of the Therapy Intensity Level Scale: Analysis of Clinimetric Properties of a Novel Approach to Assess Management of Intracranial Pressure in Traumatic Brain Injury.

    PubMed

    Zuercher, Patrick; Groen, Justus L; Aries, Marcel J H; Steyerberg, Ewout W; Maas, Andrew I R; Ercole, Ari; Menon, David K

    2016-10-01

    We aimed to assess the reliability and validity of the Therapy Intensity Level scale (TIL) for intracranial pressure (ICP) management. We reviewed the medical records of 31 patients with traumatic brain injury (TBI) in two European intensive care units (ICUs). The ICP TIL was derived over a 4-day period for 4-h (TIL4) and 24-h epochs (TIL24). TIL scores were compared with historical schemes for TIL measurement, with each other, and with clinical variables. TIL24 scores in ICU patients with TBI were compared with two control groups: patients with extracranial trauma necessitating intensive care (Trauma_ICU; n = 20) and patients with TBI not needing ICU care (TBI_WARD; n = 19), to further determine the discriminative validity of the TIL for ICP-related ICU interventions. Interrater and intraobserver agreement were excellent for TIL4 and TIL24 (Cohen κ: 0.98-0.99; intraclass correlation coefficient: 0.99-1; p < 0.0005). The mean + standard deviation (SD) TIL24 in the ICU TBI cohort was significantly higher than the Trauma_ICU patients and the TBI_WARD patients (8.2 ± 3.2 vs. 2.2 ± 0.9 and 0.1 ± 0.1, respectively; p < 0.005 for both comparisons). Correlations between the TIL scale scores and historical TIL scores, between TIL24 and the Glasgow Coma Scale, and between a range of TIL metrics and summary measures of ICP over the 4-day period, were all highly significant (p < 0.01). The results were consistent with the expected direction. A linear mixed effect analysis, accounting for within-subjects repeated measures, showed strong correlation between TIL4 and 4-h ICP (p < 0.0000005). The TIL scale is a reliable measurement instrument with a high degree of validity for assessing the therapeutic intensity level of ICP management in patients with TBI.

  4. Knowledge generating in expert systems as support of immunological investigations.

    PubMed

    Biljana, M; Jelena, M; Milorad, R; Branislav, J; Nebojsa, A

    1997-01-01

    One of the most important parameters in immunologic investigations linked with the analysis of TIL phenotypic characteristics is the ratio between particular populations and subpopulations of TIL. Values of exact parameter are not obtained with its direct measurement but with indirect measurement of values in its numerator and its denominator that represent mean values of absolute TIL figures. Exactness of estimate of specific parameter is obtained by determining the intervals of values where this parameter should lie. Determination of confidence intervals for relationship between particular populations and subpopulations of TIL was performed using Fieller's theorem which full filled requests from the nature and process of obtaining exact parameter. By analyzing the obtained confidence intervals informations can be obtained regarding TIL phenotypic characteristics of various histologic types of tumors and various types of intervals within, as well as changes in TIL content that are occurring in malignant tumors.

  5. Tumor-infiltrating lymphocyte composition, organization and PD-1/ PD-L1 expression are linked in breast cancer

    PubMed Central

    Garaud, Soizic; de Wind, Alexandre; Van den Eynden, Gert; Boisson, Anais; Gu-Trantien, Chunyan; Naveaux, Céline; Lodewyckx, Jean-Nicolas; Duvillier, Hugues; Craciun, Ligia; Veys, Isabelle; Larsimont, Denis; Piccart-Gebhart, Martine; Stagg, John; Sotiriou, Christos

    2017-01-01

    ABSTRACT The clinical relevance of tumor-infiltrating lymphocytes (TIL) in breast cancer (BC) has been clearly established by their demonstrated correlation with long-term positive outcomes. Nevertheless, the relationship between protective immunity, observed in some patients, and critical features of the infiltrate remains unresolved. This study examined TIL density, composition and organization together with PD-1 and PD-L1 expression in freshly collected and paraffin-embedded tissues from 125 patients with invasive primary BC. Tumor and normal breast tissues were analyzed using both flow cytometry and immunohistochemistry. TIL density distribution is a continuum with 25% of tumors identified as TIL-negative at a TIL density equivalent to normal breast tissues. TIL-positive tumors (75%) were equally divided into TIL-intermediate and TIL-high. Tumors had higher mean frequencies of CD4+ T cells and CD19+ B cells and a lower mean frequency of CD8+ T cells compare with normal tissues, increasing the CD4+/CD8+ T-cell ratio. Tertiary lymphoid structures (TLS), principally located in the peri-tumoral stroma, were detected in 60% of tumors and correlated with higher TIL infiltration. PD-1 and PD-L1 expression were also associated with higher TIL densities and TLS. TIL density, TLS and PD-L1 expression were correlated with more aggressive tumor characteristics, including higher proliferation and hormone receptor negativity. Our findings reveal an important relationship between PD-1/PD-L1 expression, increased CD4+ T and B-cell infiltration, TIL density and TLS, suggesting that evaluating not only the extent but also the nature and location of the immune infiltrate should be considered when evaluating antitumor immunity and the potential for benefit from immunotherapies. PMID:28197375

  6. Tumor-infiltrating lymphocyte composition, organization and PD-1/ PD-L1 expression are linked in breast cancer.

    PubMed

    Buisseret, Laurence; Garaud, Soizic; de Wind, Alexandre; Van den Eynden, Gert; Boisson, Anais; Solinas, Cinzia; Gu-Trantien, Chunyan; Naveaux, Céline; Lodewyckx, Jean-Nicolas; Duvillier, Hugues; Craciun, Ligia; Veys, Isabelle; Larsimont, Denis; Piccart-Gebhart, Martine; Stagg, John; Sotiriou, Christos; Willard-Gallo, Karen

    2017-01-01

    The clinical relevance of tumor-infiltrating lymphocytes (TIL) in breast cancer (BC) has been clearly established by their demonstrated correlation with long-term positive outcomes. Nevertheless, the relationship between protective immunity, observed in some patients, and critical features of the infiltrate remains unresolved. This study examined TIL density, composition and organization together with PD-1 and PD-L1 expression in freshly collected and paraffin-embedded tissues from 125 patients with invasive primary BC. Tumor and normal breast tissues were analyzed using both flow cytometry and immunohistochemistry. TIL density distribution is a continuum with 25% of tumors identified as TIL-negative at a TIL density equivalent to normal breast tissues. TIL-positive tumors (75%) were equally divided into TIL-intermediate and TIL-high. Tumors had higher mean frequencies of CD4(+) T cells and CD19(+) B cells and a lower mean frequency of CD8(+) T cells compare with normal tissues, increasing the CD4(+)/CD8(+) T-cell ratio. Tertiary lymphoid structures (TLS), principally located in the peri-tumoral stroma, were detected in 60% of tumors and correlated with higher TIL infiltration. PD-1 and PD-L1 expression were also associated with higher TIL densities and TLS. TIL density, TLS and PD-L1 expression were correlated with more aggressive tumor characteristics, including higher proliferation and hormone receptor negativity. Our findings reveal an important relationship between PD-1/PD-L1 expression, increased CD4(+) T and B-cell infiltration, TIL density and TLS, suggesting that evaluating not only the extent but also the nature and location of the immune infiltrate should be considered when evaluating antitumor immunity and the potential for benefit from immunotherapies.

  7. Prognostic Value of Tumor-Infiltrating Lymphocytes in Triple-Negative Breast Cancers From Two Phase III Randomized Adjuvant Breast Cancer Trials: ECOG 2197 and ECOG 1199

    PubMed Central

    Adams, Sylvia; Gray, Robert J.; Demaria, Sandra; Goldstein, Lori; Perez, Edith A.; Shulman, Lawrence N.; Martino, Silvana; Wang, Molin; Jones, Vicky E.; Saphner, Thomas J.; Wolff, Antonio C.; Wood, William C.; Davidson, Nancy E.; Sledge, George W.; Sparano, Joseph A.; Badve, Sunil S.

    2014-01-01

    Purpose Recent studies suggest that tumor-infiltrating lymphocytes (TILs) are associated with disease-free (DFS) and overall survival (OS) in operable triple-negative breast cancer (TNBC). We seek to validate the prognostic impact of TILs in primary TNBCs in two adjuvant phase III trials conducted by the Eastern Cooperative Oncology Group (ECOG). Patients and Methods Full-face hematoxylin and eosin–stained sections of 506 tumors from ECOG trials E2197 and E1199 were evaluated for density of TILs in intraepithelial (iTILs) and stromal compartments (sTILs). Patient cases of TNBC from E2197 and E1199 were randomly selected based on availability of sections. For the primary end point of DFS, association with TIL scores was determined by fitting proportional hazards models stratified on study. Secondary end points were OS and distant recurrence–free interval (DRFI). Reporting recommendations for tumor marker prognostic studies criteria were followed, and all analyses were prespecified. Results The majority of 481 evaluable cancers had TILs (sTILs, 80%; iTILs, 15%). With a median follow-up of 10.6 years, higher sTIL scores were associated with better prognosis; for every 10% increase in sTILs, a 14% reduction of risk of recurrence or death (P = .02), 18% reduction of risk of distant recurrence (P = .04), and 19% reduction of risk of death (P = .01) were observed. Multivariable analysis confirmed sTILs to be an independent prognostic marker of DFS, DRFI, and OS. Conclusion In two national randomized clinical trials using contemporary adjuvant chemotherapy, we confirm that stromal lymphocytic infiltration constitutes a robust prognostic factor in TNBCs. Studies assessing outcomes and therapeutic efficacies should consider stratification for this parameter. PMID:25071121

  8. Augmented lymphocyte expansion from solid tumors with engineered cells for costimulatory enhancement

    PubMed Central

    Friedman, Kevin M; DeVillier, Laura E; Feldman, Steven A; Rosenberg, Steven A; Dudley, Mark E

    2011-01-01

    Treatment of patients with adoptive T cell therapy requires expansion of unique tumor-infiltrating lymphocyte (TIL) cultures from single cell suspensions processed from melanoma biopsies. Strategies which increase the expansion and reliability of TIL generation from tumor digests are necessary to improve access to TIL therapy. Prior work evaluated artificial antigen presenting cells (aAPCs) for their antigen-specific and costimulatory properties. We investigated engineered cells for co-stimulatory enhancement (ECCE) consisting of K562 cells which express 4-1BBL in the absence of artificial antigen stimulation. ECCE accelerated TIL expansion and significantly improved TIL numbers (p=0.001) from single cell melanoma suspensions. TIL generated with ECCE contain significantly more CD8+CD62L+ and CD8+CD27+ T cells then comparable IL-2-expanded TIL and maintained anti-tumor reactivity. Moreover, ECCE improved TIL expansion from non-melanoma cell suspensions similar to that seen with melanoma tumors. These data demonstrate that ECCE addition to TIL production will enable treatment of patients ineligible using current methods. PMID:21989413

  9. Augmented lymphocyte expansion from solid tumors with engineered cells for costimulatory enhancement.

    PubMed

    Friedman, Kevin M; Devillier, Laura E; Feldman, Steven A; Rosenberg, Steven A; Dudley, Mark E

    2011-01-01

    Treatment of patients with adoptive T-cell therapy requires expansion of unique tumor-infiltrating lymphocyte (TIL) cultures from single-cell suspensions processed from melanoma biopsies. Strategies which increase the expansion and reliability of TIL generation from tumor digests are necessary to improve access to TIL therapy. Previous studies have evaluated artificial antigen presenting cells for their antigen-specific and costimulatory properties. We investigated engineered cells for costimulatory enhancement (ECCE) consisting of K562 cells that express 4-1BBL in the absence of artificial antigen stimulation. ECCE accelerated TIL expansion and significantly improved TIL numbers (P=0.001) from single-cell melanoma suspensions. TIL generated with ECCE contain significantly more CD8CD62L and CD8CD27 T cells then comparable interleukin-2-expanded TIL and maintained antitumor reactivity. Moreover, ECCE improved TIL expansion from nonmelanoma-cell suspensions similar to that seen with melanoma tumors. These data demonstrate that the addition of ECCE to TIL production will enable the treatment of patients that are ineligible using current methods.

  10. A Climatology of Mesoscale Wave Disturbances Seen in Mesonet Data During Storm-Fest

    DTIC Science & Technology

    2011-07-21

    requirements for the Degree of Master of Science MARINE, EARTH AND ATMOSPHERIC SCIENCES Raleigh 1996 APPROVED BY: JnYu ah Lin Ping-ung Shaw Steven E...provided the opportunity and funding required to obtain this advanced degree. Additional monetary support was provided by the COMET Outreach Project grant...dewpoint, wind speed and direction, barometric pressure, visibility , sky conditions, altimeter, density altitude and precipitation (5-min totals

  11. Aerodynamics of Supersonic Lifting Bodies

    DTIC Science & Technology

    1981-02-01

    verso of front cover. 19 Y WOROS (Continue on rt.’,;erso side i recessary and identily by block number) Theoretical Aerodynamics Lifting Bodies Wind ...waverider solution, developed from the supersonic wedge flow solution, is then i Fused to fashion vertLcal stabilizer-likh control surfaces. Wind ...served as Project Engineers ror thE wind tunnel work. Important contributions were also made bv: Mr. iis±ung Miin; Lee, -M. Beom-Soo Kim, Mtr. Martin Weeks

  12. An Investigation of the Damping in Pitch Characteristics of a Ten Degree Cone

    DTIC Science & Technology

    1975-06-01

    nm-ecure, levi grouna te Ung as currently -A.he m-c ract-lcal -way to obt-aIn aeroc vnar .2c data Teblitcrre hevnerveloclitv’ wind t unn _ie ar -he...Ballisitcs Research :ab., Aberdeen Proving Grounds, i4aryland, AIAA Paper -70, 197A 2 0 .. en,K. B., Sawyer, F. 1., Walchner, 0., "Stability Jer.vaives o.f

  13. Surface Fatigue Life of M50NiL and AISI 9310 Spur Gears and R C Bars

    DTIC Science & Technology

    1991-11-26

    AD-A241 470 NASA AVSCOM Technical Memorandum 104496 Technical Report 91- C- 034 Surface Fatigue Life of M50NiL and AISI 9310 Spur Gears and R C Bars...AISI 9310 and M50NIL In rolUng-contact ilof AISI 9310 and MSON fatigue tester. Maximum Hertz stress, 4.83 GPaFigure 4.- Typical fatigue spail (70fkA

  14. Carbon starvation increases endoglycosidase activities and production of "unconjugated N-glycans" in Silene alba cell-suspension cultures.

    PubMed Central

    Lhernould, S; Karamanos, Y; Priem, B; Morvan, H

    1994-01-01

    We previously reported the occurrence of oligomannosides and xylomannosides corresponding to unconjugated N-glycans (UNGs) in the medium of a white campion (Silene alba) cell suspension. Attention has been focused on these oligosaccharides since it was shown that they confer biological activities in plants. In an attempt to elucidate the origin of these oligosaccharides, we studied two endoglycosidase activities, putative enzymes involved in their formation. The previously described peptide-N4-(N-acetyl-glucosaminyl) asparagine amidase activity and the endo-N-acetyl-beta-D-glucosaminidase activity described in this paper were both quantified in white campion cells during the culture cycle with variable initial concentrations of sucrose. The lower the sucrose supply, the higher the two activities. Furthermore, endoglycosidase activities were greatly enhanced after the disappearance of sugar from the medium. The production of UNGs in the culture medium rose correlatively. These data strongly suggest that the production of UNGs in our white campion cell-suspension system is due to the increase of these endoglycosidase activities, which reach their highest levels of activity during conditions of carbon starvation. PMID:7991689

  15. Decreased somatic hypermutation induces an impaired peripheral B cell tolerance checkpoint

    PubMed Central

    Cantaert, Tineke; Schickel, Jean-Nicolas; Bannock, Jason M.; Ng, Yen-Shing; Massad, Christopher; Delmotte, Fabien R.; Yamakawa, Natsuko; Glauzy, Salome; Chamberlain, Nicolas; Kinnunen, Tuure; Menard, Laurence; Lavoie, Aubert; Walter, Jolan E.; Notarangelo, Luigi D.; Bruneau, Julie; Al-Herz, Waleed; Kilic, Sara Sebnem; Ochs, Hans D.; Cunningham-Rundles, Charlotte; Kuijpers, Taco W.; Kracker, Sven; Kaneko, Hideo; Nonoyama, Shigeaki; Durandy, Anne

    2016-01-01

    Patients with mutations in AICDA, which encodes activation-induced cytidine deaminase (AID), display an impaired peripheral B cell tolerance. AID mediates class-switch recombination (CSR) and somatic hypermutation (SHM) in B cells, but the mechanism by which AID prevents the accumulation of autoreactive B cells in blood is unclear. Here, we analyzed B cell tolerance in AID-deficient patients, patients with autosomal dominant AID mutations (AD-AID), asymptomatic AICDA heterozygotes (AID+/–), and patients with uracil N-glycosylase (UNG) deficiency, which impairs CSR but not SHM. The low frequency of autoreactive mature naive B cells in UNG-deficient patients resembled that of healthy subjects, revealing that impaired CSR does not interfere with the peripheral B cell tolerance checkpoint. In contrast, we observed decreased frequencies of SHM in memory B cells from AD-AID patients and AID+/– subjects, who were unable to prevent the accumulation of autoreactive mature naive B cells. In addition, the individuals with AICDA mutations, but not UNG-deficient patients, displayed Tregs with defective suppressive capacity that correlated with increases in circulating T follicular helper cells and enhanced cytokine production. We conclude that SHM, but not CSR, regulates peripheral B cell tolerance through the production of mutated antibodies that clear antigens and prevent sustained interleukin secretions that interfere with Treg function. PMID:27701145

  16. Decreased somatic hypermutation induces an impaired peripheral B cell tolerance checkpoint.

    PubMed

    Cantaert, Tineke; Schickel, Jean-Nicolas; Bannock, Jason M; Ng, Yen-Shing; Massad, Christopher; Delmotte, Fabien R; Yamakawa, Natsuko; Glauzy, Salome; Chamberlain, Nicolas; Kinnunen, Tuure; Menard, Laurence; Lavoie, Aubert; Walter, Jolan E; Notarangelo, Luigi D; Bruneau, Julie; Al-Herz, Waleed; Kilic, Sara Sebnem; Ochs, Hans D; Cunningham-Rundles, Charlotte; van der Burg, Mirjam; Kuijpers, Taco W; Kracker, Sven; Kaneko, Hideo; Sekinaka, Yujin; Nonoyama, Shigeaki; Durandy, Anne; Meffre, Eric

    2016-11-01

    Patients with mutations in AICDA, which encodes activation-induced cytidine deaminase (AID), display an impaired peripheral B cell tolerance. AID mediates class-switch recombination (CSR) and somatic hypermutation (SHM) in B cells, but the mechanism by which AID prevents the accumulation of autoreactive B cells in blood is unclear. Here, we analyzed B cell tolerance in AID-deficient patients, patients with autosomal dominant AID mutations (AD-AID), asymptomatic AICDA heterozygotes (AID+/-), and patients with uracil N-glycosylase (UNG) deficiency, which impairs CSR but not SHM. The low frequency of autoreactive mature naive B cells in UNG-deficient patients resembled that of healthy subjects, revealing that impaired CSR does not interfere with the peripheral B cell tolerance checkpoint. In contrast, we observed decreased frequencies of SHM in memory B cells from AD-AID patients and AID+/- subjects, who were unable to prevent the accumulation of autoreactive mature naive B cells. In addition, the individuals with AICDA mutations, but not UNG-deficient patients, displayed Tregs with defective suppressive capacity that correlated with increases in circulating T follicular helper cells and enhanced cytokine production. We conclude that SHM, but not CSR, regulates peripheral B cell tolerance through the production of mutated antibodies that clear antigens and prevent sustained interleukin secretions that interfere with Treg function.

  17. Impaired dynamics and function of mitochondria caused by mtDNA toxicity leads to heart failure.

    PubMed

    Lauritzen, Knut H; Kleppa, Liv; Aronsen, Jan Magnus; Eide, Lars; Carlsen, Harald; Haugen, Øyvind P; Sjaastad, Ivar; Klungland, Arne; Rasmussen, Lene Juel; Attramadal, Håvard; Storm-Mathisen, Jon; Bergersen, Linda H

    2015-08-01

    Cardiac mitochondrial dysfunction has been implicated in heart failure of diverse etiologies. Generalized mitochondrial disease also leads to cardiomyopathy with various clinical manifestations. Impaired mitochondrial homeostasis may over time, such as in the aging heart, lead to cardiac dysfunction. Mitochondrial DNA (mtDNA), close to the electron transport chain and unprotected by histones, may be a primary pathogenetic site, but this is not known. Here, we test the hypothesis that cumulative damage of cardiomyocyte mtDNA leads to cardiomyopathy and heart failure. Transgenic mice with Tet-on inducible, cardiomyocyte-specific expression of a mutant uracil-DNA glycosylase 1 (mutUNG1) were generated. The mutUNG1 is known to remove thymine in addition to uracil from the mitochondrial genome, generating apyrimidinic sites, which obstruct mtDNA function. Following induction of mutUNG1 in cardiac myocytes by administering doxycycline, the mice developed hypertrophic cardiomyopathy, leading to congestive heart failure and premature death after ∼2 mo. The heart showed reduced mtDNA replication, severely diminished mtDNA transcription, and suppressed mitochondrial respiration with increased Pgc-1α, mitochondrial mass, and antioxidative defense enzymes, and finally failing mitochondrial fission/fusion dynamics and deteriorating myocardial contractility as the mechanism of heart failure. The approach provides a model with induced cardiac-restricted mtDNA damage for investigation of mtDNA-based heart disease.

  18. Cadmium(II) inhibition of human uracil-DNA glycosylase by catalytic water supplantation

    NASA Astrophysics Data System (ADS)

    Gokey, Trevor; Hang, Bo; Guliaev, Anton B.

    2016-12-01

    Toxic metals are known to inhibit DNA repair but the underlying mechanisms of inhibition are still not fully understood. DNA repair enzymes such as human uracil-DNA glycosylase (hUNG) perform the initial step in the base excision repair (BER) pathway. In this work, we showed that cadmium [Cd(II)], a known human carcinogen, inhibited all activity of hUNG at 100 μM. Computational analyses based on 2 μs equilibrium, 1.6 μs steered molecular dynamics (SMD), and QM/MM MD determined that Cd(II) ions entered the enzyme active site and formed close contacts with both D145 and H148, effectively replacing the catalytic water normally found in this position. Geometry refinement by density functional theory (DFT) calculations showed that Cd(II) formed a tetrahedral structure with D145, P146, H148, and one water molecule. This work for the first time reports Cd(II) inhibition of hUNG which was due to replacement of the catalytic water by binding the active site D145 and H148 residues. Comparison of the proposed metal binding site to existing structural data showed that D145:H148 followed a general metal binding motif favored by Cd(II). The identified motif offered structural insights into metal inhibition of other DNA repair enzymes and glycosylases.

  19. Helix–hairpin–helix protein MJ1434 from Methanocaldococcus jannaschii and EndoIV homologue TTC0482 from Thermus thermophilus HB27 do not process DNA uracil residues

    PubMed Central

    Schomacher, Lars; Smolorz, Sabine; Ciirdaeva, Elena; Ber, Svetlana; Kramer, Wilfried; Fritz, Hans-Joachim

    2010-01-01

    The mutagenic threat of hydrolytic DNA cytosine deamination is met mostly by uracil DNA glycosylases (UDG) initiating base excision repair. However, several sequenced genomes of archaeal organisms are devoid of genes coding for homologues of the otherwise ubiquitous UDG superfamily of proteins. Previously, two possible solutions to this problem were offered by (i) a report of a newly discovered family of uracil DNA glycosylases exemplified by MJ1434, a protein found in the hyperthermophilic archaeon Methanocaldococcus jannaschii, and (ii) the description of TTC0482, an EndoIV homologue from the hyperthermophilic bacterium Thermus thermophilus HB27, as being able to excise uracil from DNA. Sequence homologues of both proteins can be found throughout the archaeal domain of life. Three proteins orthologous to MJ1434 and the family founder itself were tested for but failed to exhibit DNA uracil glycosylase activity when produced in an Ung-deficient Escherichia coli host. Likewise, no DNA uracil processing activity could be detected to be associated with TTC0482, while the protein was fully active as an AP endonuclease. We propose that the uracil processing activities formerly found were due to contaminations with Ung enzyme. Use of Δung-strains as hosts for production of putatively DNA-U processing enzymes provides a simple safeguard. PMID:20410075

  20. Genomic structural variations for cardiovascular and metabolic comorbidity

    PubMed Central

    Nazarenko, Maria S.; Sleptcov, Aleksei A.; Lebedev, Igor N.; Skryabin, Nikolay A.; Markov, Anton V.; Golubenko, Maria V.; Koroleva, Iuliia A.; Kazancev, Anton N.; Barbarash, Olga L.; Puzyrev, Valery P.

    2017-01-01

    The objective of this study was to identify genes targeted by both copy number and copy-neutral changes in the right coronary arteries in the area of advanced atherosclerotic plaques and intact internal mammary arteries derived from the same individuals with comorbid coronary artery disease and metabolic syndrome. The artery samples from 10 patients were screened for genomic imbalances using array comparative genomic hybridization. Ninety high-confidence, identical copy number variations (CNVs) were detected. We also identified eight copy-neutral changes (cn-LOHs) > 1.5 Mb in paired arterial samples in 4 of 10 individuals. The frequencies of the two gains located in the 10q24.31 (ERLIN1) and 12q24.11 (UNG, ACACB) genomic regions were evaluated in 33 paired arteries and blood samples. Two patients contained the gain in 10q24.31 (ERLIN1) and one patient contained the gain in 12q24.11 (UNG, ACACB) that affected only the blood DNA. An additional two patients harboured these CNVs in both the arteries and blood. In conclusion, we discovered and confirmed a gain of the 10q24.31 (ERLIN1) and 12q24.11 (UNG, ACACB) genomic regions in patients with coronary artery disease and metabolic comorbidity. Analysis of DNA extracted from blood indicated a possible somatic origin for these CNVs. PMID:28120895

  1. Poxvirus uracil-DNA glycosylase-An unusual member of the family I uracil-DNA glycosylases: Poxvirus Uracil-DNA Glycosylase

    SciTech Connect

    Schormann, Norbert; Zhukovskaya, Natalia; Bedwell, Gregory; Nuth, Manunya; Gillilan, Richard; Prevelige, Peter E.; Ricciardi, Robert P.; Banerjee, Surajit; Chattopadhyay, Debasish

    2016-11-02

    We report that uracil-DNA glycosylases are ubiquitous enzymes, which play a key role repairing damages in DNA and in maintaining genomic integrity by catalyzing the first step in the base excision repair pathway. Within the superfamily of uracil-DNA glycosylases family I enzymes or UNGs are specific for recognizing and removing uracil from DNA. These enzymes feature conserved structural folds, active site residues and use common motifs for DNA binding, uracil recognition and catalysis. Within this family the enzymes of poxviruses are unique and most remarkable in terms of amino acid sequences, characteristic motifs and more importantly for their novel non-enzymatic function in DNA replication. UNG of vaccinia virus, also known as D4, is the most extensively characterized UNG of the poxvirus family. D4 forms an unusual heterodimeric processivity factor by attaching to a poxvirus-specific protein A20, which also binds to the DNA polymerase E9 and recruits other proteins necessary for replication. D4 is thus integrated in the DNA polymerase complex, and its DNA-binding and DNA scanning abilities couple DNA processivity and DNA base excision repair at the replication fork. In conclusion, the adaptations necessary for taking on the new function are reflected in the amino acid sequence and the three-dimensional structure of D4. We provide an overview of the current state of the knowledge on the structure-function relationship of D4.

  2. Cadmium(II) inhibition of human uracil-DNA glycosylase by catalytic water supplantation

    PubMed Central

    Gokey, Trevor; Hang, Bo; Guliaev, Anton B.

    2016-01-01

    Toxic metals are known to inhibit DNA repair but the underlying mechanisms of inhibition are still not fully understood. DNA repair enzymes such as human uracil-DNA glycosylase (hUNG) perform the initial step in the base excision repair (BER) pathway. In this work, we showed that cadmium [Cd(II)], a known human carcinogen, inhibited all activity of hUNG at 100 μM. Computational analyses based on 2 μs equilibrium, 1.6 μs steered molecular dynamics (SMD), and QM/MM MD determined that Cd(II) ions entered the enzyme active site and formed close contacts with both D145 and H148, effectively replacing the catalytic water normally found in this position. Geometry refinement by density functional theory (DFT) calculations showed that Cd(II) formed a tetrahedral structure with D145, P146, H148, and one water molecule. This work for the first time reports Cd(II) inhibition of hUNG which was due to replacement of the catalytic water by binding the active site D145 and H148 residues. Comparison of the proposed metal binding site to existing structural data showed that D145:H148 followed a general metal binding motif favored by Cd(II). The identified motif offered structural insights into metal inhibition of other DNA repair enzymes and glycosylases. PMID:27974818

  3. The Influence of Marcellus Shale Extraction Emissions on Regionally Monitored Dry Reactive Nitrogen Deposition.

    PubMed

    Coughlin, Justin G; Rose, Lucy A; Bain, Daniel J; Elliott, Emily M

    2017-03-09

    Emissions of nitrogen oxides (NOx) in the United States (U.S.) from large stationary sources, such as electric generating units, have decreased since 1995, driving decreases in nitrogen deposition. However, increasing NOx emissions from emerging industries, such as unconventional natural gas (UNG) extraction, could offset stationary source emission reductions in shale gas producing regions of the U.S. The Marcellus Shale in the northeastern U.S. has seen dramatic increases in the number of wells and associated natural gas production during the past 10 years. In this study, we examine the potential impacts of shale gas development on regional NOx emission inventories and dry deposition fluxes to Clean Air Status and Trends (CASTNET) sites in Pennsylvania and New York. Our results demonstrate that the current distribution of CASTNET sites is ineffective for monitoring the influence of Marcellus well NOx emissions on regional nitrogen deposition. Despite the fact that existing CASTNET sites are not influenced by UNG extraction activity, NOx emissions densities from shale gas extraction are substantial and are estimated to reach up to 21 kg NOx ha(-1) year(-1) in some regions. If these emissions deposit locally, UNG extraction activity could contribute to critical nitrogen load exceedances in areas of high well density.

  4. CD8+ enriched “young” tumor infiltrating lymphocytes can mediate regression of metastatic melanoma

    PubMed Central

    Dudley, Mark E.; Gross, Colin A.; Langhan, Michelle M.; Garcia, Marcos R.; Sherry, Richard M.; Yang, James C.; Phan, Giao Q.; Kammula, Udai S.; Hughes, Marybeth S.; Citrin, Deborah E.; Restifo, Nicholas P.; Wunderlich, John; Prieto, Peter A.; Hong, Jenny J.; Langan, Russell C.; Zlott, Daniel A.; Morton, Kathleen E.; White, Donald E.; Laurencot, Carolyn; Rosenberg, Steven A.

    2010-01-01

    Purpose Tumor infiltrating lymphocytes (TIL) and interleukin (IL)-2 administered following lymphodepletion can cause the durable complete regression of bulky metastatic melanoma in patients refractory to approved treatments. However, the generation of a unique tumor-reactive TIL culture for each patient may be prohibitively difficult. We therefore investigated the clinical and immunological impact of unscreened, CD8+ enriched “young” TIL. Experimental Design Methods were developed for generating TIL that minimized the time in culture and eliminated the individualized tumor-reactivity screening step. Thirty-three patients were treated with these CD8+ enriched young TIL and IL-2 following non-myeloablative lymphodepletion (NMA). Twenty-three additional patients were treated with CD8+ enriched young TIL and IL-2 after lymphodepletion with NMA and 6Gy of total body irradiation (TBI). Results Young TIL cultures for therapy were successfully established from 83% of 122 consecutive melanoma patients. Nineteen of 33 patients (58%) treated with CD8+ enriched young TIL and NMA had an objective response (RECIST) including three complete responders. Eleven of 23 patients (48%) treated with TIL and 6Gy TBI had an objective response including two complete responders. At one month after TIL infusion the absolute CD8+ cell numbers in the periphery were highly correlated with response. Conclusion This study shows that a rapid and simplified method can be used to reliably generate CD8+ enriched young TIL for administration as an individualized therapy for advanced melanoma, and may allow this potentially effective treatment to be applied at other institutions and to reach additional patients. PMID:20668005

  5. The combination of PD-L1 expression and decreased tumor-infiltrating lymphocytes is associated with a poor prognosis in triple-negative breast cancer.

    PubMed

    Mori, Hitomi; Kubo, Makoto; Yamaguchi, Rin; Nishimura, Reiki; Osako, Tomofumi; Arima, Nobuyuki; Okumura, Yasuhiro; Okido, Masayuki; Yamada, Mai; Kai, Masaya; Kishimoto, Junji; Oda, Yoshinao; Nakamura, Masafumi

    2017-01-17

    This study included patients with primary triple-negative breast cancer (TNBC) who underwent resection without neoadjuvant chemotherapy between January 2004 and December 2014. Among the 248 TNBCs studied, programmed cell death ligand-1 (PD-L1) expression was detected in 103 (41.5%) tumors, and high levels of tumor-infiltrating lymphocytes (TILs) were present in 118 (47.6%) tumors. PD-L1 expression correlated with high levels of TILs, but was not a prognostic factor. Patients with TILs-high tumors had better overall survival than those with TILs-low tumors (P = 0.016). There was a strong interaction between PD-L1 expression and TILs that was associated with both recurrence-free survival (P = 0.0018) and overall survival (P = 0.015). Multivariate Cox proportional hazards model analysis showed that PD-L1-positive/TILs-low was an independent negative prognostic factor for both recurrence-free survival and overall survival. Our findings suggest that PD-L1-positive/TILs-low tumors are associated with a poor prognosis in patients with TNBC, and that it is important to focus on the combination of PD-L1 expression on tumor cells and TILs present in the tumor microenvironment. These biomarkers may be useful for stratification of TNBCs and for predicting prognosis and developing novel cancer immunotherapies.

  6. Tumor Infiltrating Lymphocytes Affect the Outcome of Patients with Operable Triple-Negative Breast Cancer in Combination with Mutated Amino Acid Classes

    PubMed Central

    Kotoula, Vassiliki; Lakis, Sotiris; Vlachos, Ioannis S.; Giannoulatou, Eleni; Zagouri, Flora; Alexopoulou, Zoi; Gogas, Helen; Pectasides, Dimitrios; Aravantinos, Gerasimos; Efstratiou, Ioannis; Pentheroudakis, George; Papadopoulou, Kyriaki; Chatzopoulos, Kyriakos; Papakostas, Pavlos; Sotiropoulou, Maria; Nicolaou, Irene; Razis, Evangelia; Psyrri, Amanda; Kosmidis, Paris; Papadimitriou, Christos; Fountzilas, George

    2016-01-01

    Background Stromal tumor infiltrating lymphocytes (TILs) density is an outcome predictor in triple-negative breast cancer (TNBC). Herein we asked whether TILs are related to coding mutation load and to the chemical class of the resulting mutated amino acids, i.e., charged, polar, and hydrophobic mutations. Methods We examined paraffin tumors from TNBC patients who had been treated with adjuvant chemotherapy mostly within clinical trials (training cohort, N = 133; validation, N = 190) for phenotype concordance; TILs density; mutation load and types. Results Concordance of TNBC phenotypes was 42.1% upon local / central, and 72% upon central / central pathology assessment. TILs were not associated with mutation load, type and class of mutated amino acids. Polar and charged mutation patterns differed between TP53 and PIK3CA (p<0.001). Hydrophobic mutations predicted for early relapse in patients with high nodal burden and <50% TILs tumors (training: HR 3.03, 95%CI 1.11–8.29, p = 0.031; validation: HR 2.90, 95%CI 0.97–8.70, p = 0.057), especially if compared to patients with >50% TILs tumors (training p = 0.003; validation p = 0.015). Conclusions TILs density is unrelated to mutation load in TNBC, which may be regarded as an unstable phenotype. If further validated, hydrophobic mutations along with TILs density may help identifying TNBC patients in higher risk for relapse. PMID:27685159

  7. A New Approach to the Adoptive Immunotherapy of Cancer with Tumor-Infiltrating Lymphocytes

    NASA Astrophysics Data System (ADS)

    Rosenberg, Steven A.; Spiess, Paul; Lafreniere, Rene

    1986-09-01

    The adoptive transfer of tumor-infiltrating lymphocytes (TIL) expanded in interleukin-2 (IL-2) to mice bearing micrometastases from various types of tumors showed that TIL are 50 to 100 times more effective in their therapeutic potency than are lymphokine-activated killer (LAK) cells. Therefore the use of TIL was explored for the treatment of mice with large pulmonary and hepatic metastatic tumors that do not respond to LAK cell therapy. Although treatment of animals with TIL alone or cyclophosphamide alone had little impact, these two modalities together mediated the elimination of large metastatic cancer deposits in the liver and lung. The combination of TIL and cyclophosphamide was further potentiated by the simultaneous administration of IL-2. With the combination of cyclophosphamide, TIL, and IL-2, 100% of mice (n = 12) bearing the MC-38 colon adenocarcinoma were cured of advanced hepatic metastases, and up to 50% of mice were cured of advanced pulmonary metastases. Techniques have been developed to isolate TIL from human tumors. These experiments provide a rationale for the use of TIL in the treatment of humans with advanced cancer.

  8. A Novel Trypsin Inhibitor-Like Cysteine-Rich Peptide from the Frog Lepidobatrachus laevis Containing Proteinase-Inhibiting Activity.

    PubMed

    Wang, Yu-Wei; Tan, Ji-Min; Du, Can-Wei; Luan, Ning; Yan, Xiu-Wen; Lai, Ren; Lu, Qiu-Min

    2015-08-01

    Various bio-active substances in amphibian skins play important roles in survival of the amphibians. Many protease inhibitor peptides have been identified from amphibian skins, which are supposed to negatively modulate the activity of proteases to avoid premature degradation or release of skin peptides, or to inhibit extracellular proteases produced by invading bacteria. However, there is no information on the proteinase inhibitors from the frog Lepidobatrachus laevis which is unique in South America. In this work, a cDNA encoding a novel trypsin inhibitor-like (TIL) cysteine-rich peptide was identified from the skin cDNA library of L. laevis. The 240-bp coding region encodes an 80-amino acid residue precursor protein containing 10 half-cysteines. By sequence comparison and signal peptide prediction, the precursor was predicted to release a 55-amino acid mature peptide with amino acid sequence, IRCPKDKIYKFCGSPCPPSCKDLTPNCIAVCKKGCFCRDGTVDNNHGKCVKKENC. The mature peptide was named LL-TIL. LL-TIL shares significant domain similarity with the peptides from the TIL supper family. Antimicrobial and trypsin-inhibitory abilities of recombinant LL-TIL were tested. Recombinant LL-TIL showed no antimicrobial activity, while it had trypsin-inhibiting activity with a Ki of 16.5178 μM. These results suggested there was TIL peptide with proteinase-inhibiting activity in the skin of frog L. laevis. To the best of our knowledge, this is the first report of TIL peptide from frog skin.

  9. Baroclinic mixing of potential vorticity as the principal sharpening mechanism for the extratropical Tropopause Inversion Layer

    NASA Astrophysics Data System (ADS)

    Wang, Shu Meir; Geller, Marvin A.

    2016-09-01

    Previous works have shown that a dry, idealized general circulation model could produce many features of the extratropical Tropopause Inversion Layer (TIL). In particular, the following have been shown, but no explanations were given for these results. (1) A sharper extratropical TIL resulted more from increased horizontal resolution than from increased vertical resolution. (2) If the Equator-to-Pole temperature gradient was varied, the annual variation of the extratropical TIL found in observations could be reproduced. (3) The extratropical TIL altitude showed excellent correlation with the upper tropospheric relative vorticity, as had been previously proposed. (4) Increased horizontal model resolutions led to extratropical TILs that were at lower altitudes. We show that these conclusions follow from baroclinic mixing of high stratospheric potential vorticity into the troposphere being the principal sharpening mechanism for the extratropical TIL and the increased baroclinic activity occurring in higher horizontal resolution models. We furthermore suggest that the distance from the jet exerts a greater influence on the height and sharpness of the extratropical TIL than does the upper tropospheric relative vorticity, and this accounts for the annual behavior of the extratropical TIL found in observations and reproduced with a dry, mechanistic, global model.

  10. Regulation of the Prostate Cancer Tumor Microenvironment

    DTIC Science & Technology

    2015-04-01

    are interested in understanding the mechanisms for development of TILs and how they modulate prostate cancer. Our hypothesis is that the innate ...growth can be altered through modulating the composition of TILs through innate immunity. Body Pathogens or cancerous cells alike can produce danger... innate immunity, including Toll-like receptors (TLRs). Thirteen mammalian TLRs have been identified to date with ligands ranging from

  11. Current Issues and Clinical Evidence in Tumor-Infiltrating Lymphocytes in Breast Cancer

    PubMed Central

    Ahn, Sung Gwe; Jeong, Joon; Hong, SoonWon; Jung, Woo Hee

    2015-01-01

    With the advance in personalized therapeutic strategies in patients with breast cancer, there is an increasing need for biomarker-guided therapy. Although the immunogenicity of breast cancer has not been strongly considered in research or practice, tumor-infiltrating lymphocytes (TILs) are emerging as biomarkers mediating tumor response to treatments. Earlier studies have provided evidence that the level of TILs has prognostic value and the potential for predictive value, particularly in triple-negative and human epidermal growth factor receptor 2–positive breast cancer. Moreover, the level of TILs has been associated with treatment outcome in patients undergoing neoadjuvant chemotherapy. To date, no standardized methodology for measuring TILs has been established. In this article, we review current issues and clinical evidence for the use of TILs in breast cancer. PMID:26278518

  12. Verslag Van De 15e Conferentie Over Very Large Data Bases Van 22 t/m 25 Augustus 1989 Te Amsterdam (Report on the 15th Conference on Very Large Data Bases in Amsterdam from August 22 til August 25 1989)

    DTIC Science & Technology

    1989-10-04

    8217Aunt Nellie Heuristic’ of ’Tante Nellie Methode’ (TNM): ’ Neem bet voorbeeld ter band. Ga ervan uit dat je bet voorbeeld over de telefoon moet...Un. Dortmund ’The LSD tree : spatial access to multidimensional point and non-point objects’ A. Henrich, H-W. Six and P. Widmayer; Fern Un., Hagen...USA ’Optimization and dataflow algorithms for nested tree queries’ M. Muralikrishna; DEC, Colorada Springs CO, USA ’Parallel processing of recursive

  13. Engineered artificial antigen presenting cells facilitate direct and efficient expansion of tumor infiltrating lymphocytes

    PubMed Central

    2011-01-01

    Background Development of a standardized platform for the rapid expansion of tumor-infiltrating lymphocytes (TILs) with anti-tumor function from patients with limited TIL numbers or tumor tissues challenges their clinical application. Methods To facilitate adoptive immunotherapy, we applied genetically-engineered K562 cell-based artificial antigen presenting cells (aAPCs) for the direct and rapid expansion of TILs isolated from primary cancer specimens. Results TILs outgrown in IL-2 undergo rapid, CD28-independent expansion in response to aAPC stimulation that requires provision of exogenous IL-2 cytokine support. aAPCs induce numerical expansion of TILs that is statistically similar to an established rapid expansion method at a 100-fold lower feeder cell to TIL ratio, and greater than those achievable using anti-CD3/CD28 activation beads or extended IL-2 culture. aAPC-expanded TILs undergo numerical expansion of tumor antigen-specific cells, remain amenable to secondary aAPC-based expansion, and have low CD4/CD8 ratios and FOXP3+ CD4+ cell frequencies. TILs can also be expanded directly from fresh enzyme-digested tumor specimens when pulsed with aAPCs. These "young" TILs are tumor-reactive, positively skewed in CD8+ lymphocyte composition, CD28 and CD27 expression, and contain fewer FOXP3+ T cells compared to parallel IL-2 cultures. Conclusion Genetically-enhanced aAPCs represent a standardized, "off-the-shelf" platform for the direct ex vivo expansion of TILs of suitable number, phenotype and function for use in adoptive immunotherapy. PMID:21827675

  14. TGF-β1-mediated Smad3 enhances PD-1 expression on antigen-specific T cells in cancer

    PubMed Central

    Park, Benjamin V.; Freeman, Zachary T.; Ghasemzadeh, Ali; Chattergoon, Michael A.; Rutebemberwa, Alleluiah; Steigner, Jordana; Winter, Matthew E.; Huynh, Thanh V.; Sebald, Suzanne M.; Lee, Se-Jin; Pan, Fan; Pardoll, Drew M.; Cox, Andrea L.

    2017-01-01

    Programmed Death-1 (PD-1) is a co-inhibitory receptor that down-regulates the activity of tumor-infiltrating lymphocytes (TIL) in cancer and of virus-specific T cells in chronic infection. The molecular mechanisms driving high PD-1 expression on TIL have not been fully investigated. We demonstrate that transforming growth factor-β1 (TGF-β1) directly enhances antigen-induced PD-1 expression through Smad3-dependent, Smad2-independent transcriptional activation in T cells in vitro and in TIL in vivo. The PD-1hi subset seen in CD8+ TIL is absent in Smad3-deficient tumor-specific CD8+ TIL, resulting in enhanced cytokine production by TIL and in draining lymph nodes and of anti-tumor activity. In addition to TGF-β1’s previously known effects on T cell function, our findings suggest that TGF-β1 mediates T cell suppression via PD-1 upregulation in the TME. They highlight bidirectional crosstalk between effector TIL and TGF-β-producing cells that upregulates multiple components of the PD-1 signaling pathway to inhibit anti-tumor immunity. PMID:27683557

  15. Endoplasmic reticulum stress induces secretion of high-mobility group proteins and is associated with tumor-infiltrating lymphocytes in triple-negative breast cancer

    PubMed Central

    Park, In Ah; Heo, Sun-Hee; Song, In Hye; Kim, Young-Ae; Park, Hye Seon; Bang, Won Seon; Park, Suk Young; Jo, Jeong-Hyon; Lee, Hee Jin; Gong, Gyungyub

    2016-01-01

    Background Although the prognostic and predictive significance of tumor-infiltrating lymphocytes (TILs) in triple-negative breast cancer (TNBC) have been shown, the cause of the TIL influx is unclear. Here, we investigated whether extracellular secretion of HMGN1 is associated with TIL influx, as well as increased endoplasmic reticulum stress (ERS), in human TNBC. Methods We reviewed the slides of 767 patients with TNBC and evaluated the TIL levels. We also assessed the expression of HMGs and several ERS-associated molecules using immunohistochemical staining. Western blot analysis of human TNBC cell lines and pharmacological ERS inducers was used to determine if HMGN1 migrates from the nucleus to the extracellular space in response to ERS. Results On immunohistochemical staining, either higher nuclear or cytoplasmic expression of both HMGB1 and HMGN1 was significantly associated with ERS. TILs showed a positive correlation with the cytoplasmic expression of the HMGs. Western blot analysis of TNBC cell lines showed that ERS induction resulted in the secretion of HMG proteins. Conclusions This is the first study to elucidate the associations among ERS, secretion of HMGs, and degree of TILs in TNBCs. Understanding the mechanisms of TIL influx will help in the development of effective immunotherapeutic agents for TNBC. PMID:27494867

  16. Modulating Effects of Spirulina platensis against Tilmicosin-Induced Cardiotoxicity in Mice

    PubMed Central

    Ibrahim, Abdelaziz E.; Abdel-Daim, Mohamed Mohamed

    2015-01-01

    Objective Tilmicosin (TIL) is a long-acting macrolide antibiotic used to treat cattle for pathogens that cause bovine respiratory disease. However, overdoses of this medication have been reported to induce cardiac damage. Our experimental objective was to evaluate the protective effects of Spirulina platensis (SP) administration against TIL-induced cardiotoxicity in mice. Materials and Methods Our experimental in vivo animal study used 40 male albino mice that were divided into five groups of eight mice per group. The first group served as a control group and was injected with saline. The second group received SP at dose of 1000 mg/kg body weight for five days. The third group received a single dose of TIL (75 mg/kg, subcutaneously). Groups 4 and 5 were given SP at doses of 500 and 1000 mg/kg body weight for five consecutive days just before administration of TIL at the same dose and regimen used for group 3. Results TIL treated animals showed a significant increase in serum cardiac injury biomarkers as well as cardiac lipid peroxidation, however they had evidence of an inhibition in antioxidant biomarkers. SP normalized elevated serum levels of lactate dehydrogenase (LDH), creatine kinase (CK), and CK-MB. Furthermore, SP reduced TIL-induced lipid peroxidation and oxidative stress in a dose-dependent manner. Conclusion Administration of SP minimized the toxic effects of TIL by its free radicalscavenging and potent antioxidant activity. PMID:25870843

  17. Synergistic protective role of mirazid (Commiphora molmol) and ascorbic acid against tilmicosin-induced cardiotoxicity in mice.

    PubMed

    Abdel-Daim, Mohamed M; Ghazy, Emad W; Fayez, Mostafa

    2015-01-01

    Tilmicosin (TIL) is a long-acting macrolide antibiotic approved for the treatment of cattle with Bovine Respiratory Disease. However, overdose of TIL has been reported to induce cardiotoxicity. The purpose of our experiment was to evaluate the protective effects of Commiphora molmol (mirazid (MRZ); myrrh) and (or) ascorbic acid (AA) against TIL-induced cardiotoxicity in mice. MRZ and AA were orally administered using stomach gavage, either alone or in combination for 5 consecutive days, followed with a single TIL overdose. TIL overdose induced a significant increase in serum levels of cardiac damage biomarkers (AST, LDH, CK, CK-MB, and cTnT), as well as cardiac lipid peroxidation, but cardiac levels of antioxidant biomarkers (GSH, SOD, CAT, and TAC) were decreased. Both MRZ and AA tended to normalize the elevated serum levels of cardiac injury biomarkers. Furthermore, MRZ and AA reduced TIL-induced lipid peroxidation and oxidative stress parameters. MRZ and AA combined produced a synergistic cardioprotective effect. We conclude that myrrh and (or) vitamin C administration minimizes the toxic effects of TIL through their free-radical-scavenging and potent antioxidant activities.

  18. Glutaminase expression is a poor prognostic factor in node-positive triple-negative breast cancer patients with a high level of tumor-infiltrating lymphocytes.

    PubMed

    Kim, Joo Young; Heo, Sun-Hee; Choi, Seul Ki; Song, In Hye; Park, In Ah; Kim, Young-Ae; Park, Hye Seon; Park, Suk Young; Bang, Won Seon; Gong, Gyungyub; Lee, Hee Jin

    2017-04-01

    Glutamine metabolism is emerging as one aspect of dysregulated metabolism of tumors. Triple-negative breast cancer (TNBC) cells are glutamine dependent, whereas luminal-type cells tend to be glutamine independent. Therefore, TNBC patients might benefit from therapies targeting glutamine metabolism. To investigate the clinical significance of glutamine metabolism, we examined expression and prognostic significance of glutaminase in tumor cells and tumor-infiltrating lymphocytes (TILs) in TNBC. We retrieved 658 surgically resected TNBCs and analyzed glutaminase expression in tumor cells and TILs by immunohistochemical staining. Glutaminase expression was observed in 237 cases (36.0%) in tumor cells and 104 cases (15.5%) in TILs. Although glutaminase expression in tumor cells was significantly associated with a low level of TILs (p = 0.018), glutaminase expression in TILs was significantly higher in cases with a high level of TILs (p = 0.031). Glutaminase expression in tumor cells was significantly associated with poor disease-free survival in patients with lymph node metastasis and high levels of TILs (p = 0.020). In addition, it was an independent poor prognostic factor (hazard ratio = 10.643, 95% confidence interval = 1.999-56.668; p = 0.006). Glutaminase expression in tumor cells was observed in a subset of TNBC patients. It was significantly associated with a low level of TILs and poor disease-free survival in TNBCs presenting with lymph node metastasis and high levels of TILs.

  19. Phenotype and function of T cells infiltrating visceral metastases from gastrointestinal cancers and melanoma: implications for adoptive cell transfer therapy.

    PubMed

    Turcotte, Simon; Gros, Alena; Hogan, Katherine; Tran, Eric; Hinrichs, Christian S; Wunderlich, John R; Dudley, Mark E; Rosenberg, Steven A

    2013-09-01

    Adoptive cell transfer of tumor-infiltrating lymphocytes (TILs) can mediate cancer regression in patients with metastatic melanoma, but whether this approach can be applied to common epithelial malignancies remains unclear. In this study, we compared the phenotype and function of TILs derived from liver and lung metastases from patients with gastrointestinal (GI) cancers (n = 14) or melanoma (n = 42). Fewer CD3(+) T cells were found to infiltrate GI compared with melanoma metastases, but the proportions of CD8(+) cells, T cell differentiation stage, and expression of costimulatory molecules were similar for both tumor types. Clinical-scale expansion up to ~50 × 10(9) T cells on average was obtained for all patients with GI cancer and melanoma. From GI tumors, however, TIL outgrowth in high-dose IL-2 yielded 22 ± 1.4% CD3(+)CD8(+) cells compared with 63 ± 2.4% from melanoma (p < 0.001). IFN-γ ELISA demonstrated MHC class I-mediated reactivity of TIL against autologous tumor in 5 of 7 GI cancer patients tested (9% of 188 distinct TIL cultures) and in 9 of 10 melanoma patients (43% of 246 distinct TIL cultures). In these assays, MHC class I-mediated up-regulation of CD137 (4-1BB) expression on CD8(+) cells suggested that 0-3% of TILs expanded from GI cancer metastases were tumor-reactive. This study implies that the main challenge to the development of TIL adoptive cell transfer for metastatic GI cancers may not be the in vitro expansion of bulk TILs, but the ability to select and enrich for tumor-reactive T cells.

  20. Tumor-infiltrating lymphocytes in breast cancer predict the response to chemotherapy and survival outcome: A meta-analysis

    PubMed Central

    Wang, Ke; Xu, Jianjun; Zhang, Tao; Xue, Dan

    2016-01-01

    Tumor-infiltrating lymphocytes (TILs) influence tumor prognosis and the chemotherapeutic response. Here, we quantified the clinical relevance of TILs, including the effect of TILs on lymphocyte subpopulations and assessed their consistency in breast cancer. We searched published literature from January 2000 to January 2016. The main parameters analyzed were pathological complete response (pCR) and survival outcome following chemotherapy in patients with breast cancer. Pooled odds ratio (OR) or relative risk (RR) values with 95% confidence intervals (CIs) were computed using random and fixed-effects models. Subgroup and heterogeneity analyses were also conducted. Twenty-three studies, which included 13,100 patients, met the inclusion criteria. The pooled results showed that TILs were associated with clinicopathological parameters of biologically aggressive phenotypes, such as high tumor grade or estrogen/progesterone receptor negativity, but they were not correlated with human epidermal growth factor receptor-2 expression. Moreover, a high TIL level was associated with a significantly improved pCR rate compared with a low TIL level (OR, 2.81; P < 0.001), particularly in the triple-negative breast cancer subtype (OR, 4.67; P < 0.001). An analysis of lymphocyte subpopulations showed that infiltration by CD8 lymphocytes, but not by CD4 lymphocytes and Foxp3 cells, was associated with a high pCR rate. Furthermore, a high TIL level was associated with significantly longer disease-free survival and overall survival. Our present meta-analysis indicates that an increased number of TILs predicted pCR to chemotherapy and improved survival. A high TIL level, characterized mainly by the infiltration of CD8 lymphocytes, is a strong predictive and prognostic factor. PMID:27329588

  1. Antigen(s)-specific tumour-infiltrating lymphocytes from tumour induced by human herpes virus-6 (HHV-6) DNA transfected NIH 3T3 transformants.

    PubMed Central

    Puri, R K; Leland, P; Razzaque, A

    1991-01-01

    Tumour infiltrating lymphocytes (TIL) have recently been shown to mediate potent therapeutic effects in certain malignancies in mice and in humans. To understand the mechanism of TIL immunotherapy it would be advantageous to generate tumour-specific TIL and to study a defined system of TIL and target cells in which the tumour epitope(s) recognized by TIL might be identified. We have established tumourigenic cell lines by transfection of NIH 3T3 cells with the entire genome of human herpesvirus-6 (HHV-6) and its small fragment (about 5% of the viral DNA sequence). Injection of these cells into nude mice produced tumours termed G-2T and 14-2T, respectively. Cell lines derived from these tumours when injected in NIH Swiss mice produced tumours, G-2TS and 14-2TS, respectively. We have generated TIL from G-2TS tumour that can kill G-2TS tumour cells in vitro but not other related tumours (14-2TS or MCA-106). These TIL can be expanded between 2-6.5 every 3-5 days. The TIL proliferated in tissue culture in response to recombinant interleukin-2 and interleukin-4 and maintained their tumor specificity for up to 6 months in vitro. Their phenotype was Thy 1.2+, Lyt-2+ and L3T4-. The availability of such tumour-specific stable TIL lines and specific viral-transformed targets will provide an opportunity to characterize the tumour-associated antigen critical for the specific cytotoxicity in this system and thereby to clarify the mechanism of this promising immunological approach to cancer therapy. Images Fig. 1 PMID:1703057

  2. The effects of granulocyte-macrophage colony-stimulating factor on tumour-infiltrating lymphocytes from renal cell carcinoma.

    PubMed Central

    Steger, G. G.; Kaboo, R.; deKernion, J. B.; Figlin, R.; Belldegrun, A.

    1995-01-01

    It has been shown that granulocyte-macrophage colony-stimulating factor (GM-CSF) can induce specific and non-specific anti-tumour cytotoxicity and also stimulates the proliferation and function of peripheral lymphocytes and thymocytes. GM-CSF and interleukin 2 (IL-2) act synergistically on peripheral lymphocytes for the induction of a highly effective cytotoxic cell population. Thus, the goal of our investigation was to study the effects of GM-CSF upon expansion, proliferation and in vitro killing activity of tumour-infiltrating lymphocytes (TILs) from renal cell carcinoma (RCC). TILs from seven consecutive tumours were cultured with GM-CSF (500 or 1000 nmol ml-1) without IL-2 supplementation, with suboptimal doses of IL-2 (8 and 40 U ml-1) plus GM-CSF (1000 nmol ml-1), and with a dose of IL-2 (400 U ml-1) which sufficed alone to induce TIL development plus GM-CSF (500 or 1000 nmol ml-1). GM-CSF alone or together with suboptimal doses of IL-2 was not able to induce or facilitate TIL development in these cultures. When GM-CSF at both concentrations studied was added to optimal doses of IL-2 the resulting TIL populations proliferated significantly better and faster (+66%), resulting in a higher cell yield (+24%) at the time of maximal expansion of the TIL cultures. The length of the culture periods of TILs was not affected by GM-CSF when compared with the control cultures supplemented with IL-2 alone. In vitro killing activity of TIL populations stimulated with IL-2 and GM-CSF remained unspecific, but lysis of the autologous tumour targets as well as the allogeneic renal tumour targets was significantly enhanced (+138%) as compared with the corresponding control TILs stimulated with IL-2 alone. Lysis of the natural killer (NK)-sensitive control cell line K562 and the NK-resistant Daudi cell line remained unchanged even though FACS analysis of TILs cultured with IL-2 and 1000 nmol of GM-CSF demonstrated a significantly higher proportion of cells expressing the CD56

  3. Holistic Debriefing: A Paradigm Shift in Leadership

    DTIC Science & Technology

    2009-04-01

    Officer Rolf Folland for the RNOAF 330 Squadron on the subject “debriefing”, 8 October 2005. 2 Bjartveit and Kjærstad, Fra kaos til kosmos , 17 3... kosmos , 22 25 Ibid, 23 26 Moxnes, Hverdagens Angst, 194 27 Bjartveit and Kjærstad, Fra kaos til kosmos , 17 28 Useem, The Leadership Moment, 60 29 Ibid...Psychology (9. ed.). Orlando, FL: Harcourt Brace Jovanovich Publishers. 1985. Bjartveit, S. & Kjærstad, T. Fra kaos til kosmos . Oslo: Oslo Kolle Forlag

  4. Pathophysiology of B-cell intrinsic immunoglobulin class switch recombination deficiencies.

    PubMed

    Durandy, Anne; Taubenheim, Nadine; Peron, Sophie; Fischer, Alain

    2007-01-01

    B-cell intrinsic immunoglobulin class switch recombination (Ig-CSR) deficiencies, previously termed hyper-IgM syndromes, are genetically determined conditions characterized by normal or elevated serum IgM levels and an absence or very low levels of IgG, IgA, and IgE. As a function of the molecular mechanism, the defective CSR is variably associated to a defect in the generation of somatic hypermutations (SHMs) in the Ig variable region. The study of Ig-CSR deficiencies contributed to a better delineation of the mechanisms underlying CSR and SHM, the major events of antigen-triggered antibody maturation. Four Ig-CSR deficiency phenotypes have been so far reported: the description of the activation-induced cytidine deaminase (AID) deficiency (Ig-CSR deficiency 1), caused by recessive mutations of AICDA gene, characterized by a defect in CSR and SHM, clearly established the role of AID in the induction of the Ig gene rearrangements underlying CSR and SHM. A CSR-specific function of AID has, however, been detected by the observation of a selective CSR defect caused by mutations affecting the C-terminus of AID. Ig-CSR deficiency 2 is the consequence of uracil-N-glycosylase (UNG) deficiency. Because UNG, a molecule of the base excision repair machinery, removes uracils from DNA and AID deaminates cytosines into uracils, that observation indicates that the AID-UNG pathway directly targets DNA of switch regions from the Ig heavy-chain locus to induce the CSR process. Ig-CSR deficiencies 3 and 4 are characterized by a selective CSR defect resulting from blocks at distinct steps of CSR. A further understanding of the CSR machinery is expected from their molecular definition.

  5. Bibliography on Cold Regions Science and Technology. Cumulative Subject Index. Volumes 38-42

    DTIC Science & Technology

    1988-12-01

    TrivclUng in intarctic weither. Junqucin Villeli , R., [1986, p.42-S5, porj 41-517 Iceberg si|htingi during SiBEX-2, Chile, in Brtiufleld 41.M12... urbanised territories of northern West Siberia. Antonov-Druzhinin. V,p., ct al, [1983, p,14l-l42. rusj 38-4516 Heat and moisture transfer in frost...structure of D20 ice VII. Jorgensen, J.D., et al. (1983, p.329-333, eng] 39-3977 Far-infrared spectrum of ice Vill . Tay, S.P., et al, [1983

  6. Army Recruiting Resourcing System.

    DTIC Science & Technology

    1990-09-01

    Sheridan, Illinois 60037-6090 REPORT DOCUMENTATION PAGE OMO No. 070虨 I.& Mrw ung U btOfl tOf thti (CM .; 9 Of nOMToMt,0* Mt e,,tiSO w &rIK" 1 OUr...are those of the author and should not be construed as an official Department of the Army policy or position, unless so designated by other...changes in policies , regulations, or statutes that affect recruiting. Those interviewed within the DA staff (DCSPER, PA&E, and DAB personnel) believe there

  7. Republication of: The redshift of extragalactic nebulae

    NASA Astrophysics Data System (ADS)

    Zwicky, F.

    2009-01-01

    This English translation of the paper by F. Zwicky, "Die Rotverschieb ung von extragalaktischen Nebeln", Helv. Phys. Acta 6, 110-127 (1933), in which he concluded from the results of observations that the amount of non-luminous matter in the Universe must be greater than that of luminous matter -- thus becoming an early precursor of the dark matter idea, has been selected for publication in the Golden Oldies series of General Relativity and Gravitation. The paper is accompanied by an editorial note written by Jürgen Ehlers and by Zwicky's brief biography compiled by Andrzej Krasiński.

  8. Transient Combustion in Granular Propellant Beds. Part I. Theoretical Modeling and Numerical Solution of Transient Combustion Processes in Mobile Granular Propellant Beds

    DTIC Science & Technology

    1977-08-01

    both phases. Some bulk properties of the fluid defined in the formulati•r. and the gross effects of heat transfer and flow resistance can be measured...34adrstandUng of highb speed f3w.Jm )propspa:1.2 ad tie Lkso*m~ bt%./’Yor of t-,Aphase ruacti:ve flow 2y1v:te; Ma6 Q2 to *e4W.g tiw ~ratqw of pas...empirical input such as heat transfer, flow resistance, Intergranular stress and the burning rate law. They are found to be of hyperbolic type, since all

  9. Chinese-English Automation and Computer Technology Dictionary, Volume 2.

    DTIC Science & Technology

    1980-08-01

    chfian juan ch’ uang chuang chang zhang chtleh jue di’ui chui chao zhao chn jun ch’un chun che he ch’a cha ch ’ung chung chen zhen ch’ai chai ch’tl qu...cheng zheng ch’an chan ch’tan quan chi ji chlang chang ch’deh que ddia jia ch’ao chao rd-’fn qun chiang jiang ch’e che E chiao jiao ch’en chen e (o) e...cidai Mylar tape 06 .fun N. od alcohol; methyl 07 aloohol ( chen .) ,3 inu , wood alcohol; wood sririt; 08 7Lethy] alcohol (chem.) mxubiac target

  10. The Rolling Resistance and Sinkage of Towed Dual Wheel Combination in Sand

    DTIC Science & Technology

    1984-08-01

    Prell Ion of Pneumat 1 Tyres on San 1 AD-P004 263 Efferts of Slip on Rflergy Distribution between Tyre ani Soil AJ-P004 2f...Mo leli sat lo.i IPS I’,-US hors Routes et 1u Sol en Vue de 1’AmoUor.-t Ion " la Tra 11 on (MoielUng of Off-Roai Tyre « an... Tyre Performance AO-POO’« 261 The Rolling Resist n:v an’ Sinkage of Tow.-d Dual Wheel Combinations In Sril AD-POO’* 262 Performan

  11. International Conference on Photochemistry (15th) Abstracts of Invited Lectures and Oral and Poster Contributions Held in Paris, France on 28 July-2 August 1991 (XVeme Conference Internationale de Photochimie, Paris, France 28 Juillet-2 Aout 1991)

    DTIC Science & Technology

    1991-08-01

    RICIHARD and Pierre BDOULC Laboratolre de Photochimie 1ol6culalre et Hacromolculalire, Uh CHRS 433, Universit Blaise Pascal (Clermont-Ferrand), F...PROCESSES IN SATURAiLE DYES STUDIED BY TIME RESOLVED SPECTROSCOPY Monique M. MARTIN, Pascal PLAZA, Nguyen DAI HUNG, Yves i1. MEYER Laboratoire dc...CONFOIINATIONNELLEH ULT’RARAPI)E D)ES COLORANTS SATURAIILES ET’UIEI P’AR SPEVUEROSCOPIE RESOLUI’ EN TEMI’S Monique M. MAirIN, Pascal PLAZA, Nguyen DAI I UNG

  12. Stress and Hair Loss: Are They Related?

    MedlinePlus

    ... hair. Trichotillomania. Trichotillomania (trik-o-til-o-MAY-nee-uh) is an irresistible urge to pull out ... Flavin, M.D. References Shapiro J, et al. Evaluation and diagnosis of hair loss. http://www.uptodate. ...

  13. Improved Personalized Cancer Immunotherapy: Rapid Selection of Tumor-Reactive T Cells based on Expression of Specific Cell Surface Markers | NCI Technology Transfer Center | TTC

    Cancer.gov

    The National Cancer Institute’s Surgery Branch seeks partners interested in collaborative research to co-develop adoptive transfer of tumor infiltrating leukocytes (TIL) for cancers other than melanoma.

  14. Tumor infiltrating lymphocytes in triple negative breast cancer receiving neoadjuvant chemotherapy

    PubMed Central

    Castaneda, Carlos A; Mittendorf, Elizabeth; Casavilca, Sandro; Wu, Yun; Castillo, Miluska; Arboleda, Patricia; Nunez, Teresa; Guerra, Henry; Barrionuevo, Carlos; Dolores-Cerna, Ketty; Belmar-Lopez, Carolina; Abugattas, Julio; Calderon, Gabriela; De La Cruz, Miguel; Cotrina, Manuel; Dunstan, Jorge; Gomez, Henry L; Vidaurre, Tatiana

    2016-01-01

    AIM To determine influence of neoadjuvant-chemotherapy (NAC) over tumor-infiltrating-lymphocytes (TIL) in triple-negative-breast-cancer (TNBC). METHODS TILs were evaluated in 98 TNBC cases who came to Instituto Nacional de Enfermedades Neoplasicas from 2005 to 2010. Immunohistochemistry staining for CD3, CD4, CD8 and FOXP3 was performed in tissue microarrays (TMA) sections. Evaluation of H/E in full-face and immunohistochemistry in TMA sections was performed in pre and post-NAC samples. STATA software was used and P value < 0.05 was considered statistically significant. RESULTS Higher TIL evaluated in full-face sections from pre-NAC tumors was associated to pathologic-complete-response (pCR) (P = 0.0251) and outcome (P = 0.0334). TIL evaluated in TMA sections showed low level of agreement with full-face sections (ICC = 0.017-0.20) and was not associated to pCR or outcome. TIL in post-NAC samples were not associated to response or outcome. Post-NAC lesions with pCR had similar TIL levels than those without pCR (P = 0.6331). NAC produced a TIL decrease in full-face sections (P < 0.0001). Percentage of TIL subpopulations was correlated with their absolute counts. Higher counts of CD3, CD4, CD8 and FOXP3 in pre-NAC samples had longer disease-free-survival (DFS). Higher counts of CD3 in pre-NAC samples had longer overall-survival. Higher ratio of CD8/CD4 counts in pre-NAC was associated with pCR. Higher ratio of CD4/FOXP3 counts in pre-NAC was associated with longer DFS. Higher counts of CD4 in post-NAC samples were associated with pCR. CONCLUSION TIL in pre-NAC full-face sections in TNBC are correlated to longer survival. TIL in full-face differ from TMA sections, absolute count and percentage analysis of TIL subpopulation closely related. PMID:27777881

  15. Subverting the adaptive immune resistance mechanism to improve clinical responses to immune checkpoint blockade therapy

    PubMed Central

    Kim, Young J

    2015-01-01

    The correlation between tumor-infiltrating lymphocyte (TIL)-expression of programmed cell death ligand 1 (PD-L1) and clinical responsiveness to the PD-1 blocking antibody nivolumab implicates adaptive immune evasion mechanisms in cancer. We review our findings that tumor cell PD-L1 expression is induced by interferon γ (IFNγ) producing TILs. We provide a mechanistic rationale for combining IFNγ+ T helper type 1 (Th1)-inducing cancer vaccines with PD-1 immune checkpoint blockade. PMID:25964860

  16. Specific lymphocyte subsets predict response to adoptive cell therapy using expanded autologous tumor-infiltrating lymphocytes in metastatic melanoma patients

    PubMed Central

    Radvanyi, Laszlo G.; Bernatchez, Chantale; Zhang, Minying; Fox, Patricia S.; Miller, Priscilla; Chacon, Jessica; Wu, Richard; Lizee, Gregory; Mahoney, Sandy; Alvarado, Gladys; Glass, Michelle; Johnson, Valen E.; McMannis, John D.; Shpall, Elizabeth; Prieto, Victor; Papadopoulos, Nicholas; Kim, Kevin; Homsi, Jade; Bedikian, Agop; Hwu, Wen-Jen; Patel, Sapna; Ross, Merrick I.; Lee, Jeffrey E.; Gershenwald, Jeffrey E.; Lucci, Anthony; Royal, Richard; Cormier, Janice N.; Davies, Michael A.; Mansaray, Rahmatu; Fulbright, Orenthial J.; Toth, Christopher; Ramachandran, Renjith; Wardell, Seth; Gonzalez, Audrey; Hwu, Patrick

    2012-01-01

    Purpose Adoptive cell therapy (ACT) using autologous tumor-infiltrating lymphocytes (TIL) is a promising treatment for metastatic melanoma unresponsive to conventional therapies. We report here on the results of an ongoing Phase II clinical trial testing the efficacy of ACT using TIL in metastatic melanoma patients and the association of specific patient clinical characteristics and the phenotypic attributes of the infused TIL with clinical response. Experimental Design Altogether, 31 transiently lymphodepleted patients were treated with their expanded TIL followed by two cycles of high-dose (HD) IL-2 therapy. The effects of patient clinical features and the phenotypes of the T-cells infused on clinical response were determined. Results Overall, 15/31 (48.4%) patients had an objective clinical response using immune-related response criteria (irRC), with two patients (6.5%) having a complete response. Progression-free survival of >12 months was observed for 9/15 (60%) of the responding patients. Factors significantly associated with objective tumor regression included a higher number of TIL infused, a higher proportion of CD8+ T-cells in the infusion product, a more differentiated effector phenotype of the CD8+ population and a higher frequency of CD8+ T-cells co-expressing the negative costimulation molecule “B- and T-lymphocyte attenuator” (BTLA). No significant difference in telomere lengths of TIL between responders and non-responders was identified. Conclusion These results indicate that immunotherapy with expanded autologous TIL is capable of achieving durable clinical responses in metastatic melanoma patients and that CD8+ T-cells in the infused TIL, particularly differentiated effectors cells and cells expressing BTLA, are associated with tumor regression. PMID:23032743

  17. Co-stimulation through the CD137/4-1BB pathway protects human melanoma tumor-infiltrating lymphocytes from activation-induced cell death and enhances anti-tumor effector function

    PubMed Central

    Hernandez-Chacon, Jessica Ann; Li, Yufeng; Wu, Richard C.; Bernatchez, Chantale; Wang, Yijun; Weber, Jeffrey; Hwu, Patrick; Radvanyi, Laszlo

    2011-01-01

    Adoptive T-cell therapy (ACT) using expanded tumor-infiltrating lymphocytes (TIL) with high-dose IL-2 is a promising form of immunotherapy for Stage IV melanoma having clinical response rates of 50% or more. One of the major problems preventing further success of this therapy is that the current protocols used to highly expand TIL for infusion drive CD8+ T cells to differentiate into effector cells losing key co-stimulatory molecules such as CD28 and CD27. This has been associated with a lack of persistence in vivo for reasons not entirely clear. In this study, we demonstrate that while human melanoma CD8+ TIL lost CD27 and CD28 expression during the rapid expansion for ACT, they gained expression of the alternative co-stimulatory molecule CD137/4-1BB, and to a lesser extent CD134/OX40. Post-REP TIL were found to be highly sensitive to activation-induced cell death (AICD) when re-activated through the TCR with low levels of OKT3 antibody. However, co-ligation of 4-1BB using two different agonistic anti-4-1BB antibodies potently prevented AICD of post-REP CD8+ TIL, including those specific for MART-1, and facilitated even further cell expansion. This was correlated with increased levels of bcl-2 and bcl-xL together with decreased bim expression. 4-1BB-co-stimulated post-REP TIL also expressed increased levels of the cytolytic granule proteins and exhibited enhanced CTL activity against melanoma cells. Lastly, post-REP CD8+ TIL were protected from cell death by anti-4-1BB ligation when exposed to HLA-matched melanoma cells. Our results indicate that 4-1BB co-stimulation may significantly improve TIL survival during melanoma ACT and boost anti-tumor cytolytic activity. PMID:21389874

  18. Adaptive resistance to anti-PD1 therapy by Tim-3 upregulation is mediated by the PI3K-Akt pathway in head and neck cancer.

    PubMed

    Shayan, Gulidanna; Srivastava, Raghvendra; Li, Jing; Schmitt, Nicole; Kane, Lawrence P; Ferris, Robert L

    2017-01-01

    Programmed Death 1 (PD-1) and T cell Ig and mucin domain-3 protein (Tim-3) are immune checkpoint receptors that are expressed on tumor-infiltrating lymphocytes (TIL) in tumor-bearing mice and humans. As anti-PD-1 single agent response rates are only <20% in head and neck squamous cell carcinoma (HNSCC) patients, it is important to understand how multiple inhibitory checkpoint receptors maintain suppressed cellular immunity. One such receptor, Tim-3, activates downstream proliferative pathways through Akt/S6, and is highly expressed in dysfunctional TIL. We observed that PD-1 and Tim-3 co-expression was associated with a more exhausted phenotype, with the highest PD-1 levels on TIL co-expressing Tim-3. Dampened Akt/S6 phosphorylation in these PD-1(+)Tim-3(+) TIL, when the PD-1 pathway was ligated, suggested that signaling cross-talk could lead to escape through Tim-3 expression. Indeed, PD-1 blockade of human HNSCC TIL led to further Tim-3 upregulation, supporting a circuit of compensatory signaling and potentially permitting escape from anti-PD-1 blockade in the tumor microenvironment. Also, in a murine HNC tumor model that is partially responsive to anti-PD-1 therapy, Tim-3 was upregulated in TIL from persistently growing tumors. Significant antitumor activity was observed after sequential addition of anti-Tim-3 mAb to overcome adaptive resistance to anti-PD-1 mAb. This increased Tim-3-mediated escape of exhausted TIL from PD-1 inhibition that was mediated by phospho-inositol-3 kinase (PI3K)/Akt complex downstream of TCR signaling but not cytokine-mediated pathways. Taken together, we conclude that during PD-1 blockade, TIL upregulate Tim-3 in a PI3K/Akt-dependent manner, providing further support for dual targeting of these molecules for more effective cancer immunotherapy.

  19. Identification of Tumor Rejection Antigens for Breast Cancer Using a Mouse Tumor Rejection Model

    DTIC Science & Technology

    2009-05-01

    high throughput antigen discovery tools have been developed that have greatly helped the identification of immunogenic proteins in breast cancer...streptomycin and L-glutamine. T cell enrichment Tumor-infiltrating lymphocytes (TIL) were harvested by mincing the tu- mor and screening. The TIL were...kinase 1. Multiple proteins involved in the Rho/Rho-associated, coiled coil–containing protein kinase (Rock) signal transduction pathway were found to

  20. Up-regulation of cerebral cytochrome-c-oxidase and hemodynamics by transcranial infrared laser stimulation: A broadband near-infrared spectroscopy study.

    PubMed

    Wang, Xinlong; Tian, Fenghua; Reddy, Divya D; Nalawade, Sahil S; Barrett, Douglas W; Gonzalez-Lima, Francisco; Liu, Hanli

    2017-01-01

    Transcranial infrared laser stimulation (TILS) is a noninvasive form of brain photobiomulation. Cytochrome-c-oxidase (CCO), the terminal enzyme in the mitochondrial electron transport chain, is hypothesized to be the primary intracellular photoacceptor. We hypothesized that TILS up-regulates cerebral CCO and causes hemodynamic changes. We delivered 1064-nm laser stimulation to the forehead of healthy participants ( n = 11), while broadband near-infrared spectroscopy was utilized to acquire light reflectance from the TILS-treated cortical region before, during, and after TILS. Placebo experiments were also performed for accurate comparison. Time course of spectroscopic readings were analyzed and fitted to the modified Beer-Lambert law. With respect to the placebo readings, we observed (1) significant increases in cerebral concentrations of oxidized CCO (Δ[CCO]; >0.08 µM; p < 0.01), oxygenated hemoglobin (Δ[HbO]; >0.8 µM; p < 0.01), and total hemoglobin (Δ[HbT]; >0.5 µM; p < 0.01) during and after TILS, and (2) linear interplays between Δ[CCO] versus Δ[HbO] and between Δ[CCO] versus Δ[HbT]. Ratios of Δ[CCO]/Δ[HbO] and Δ[CCO]/Δ[HbT] were introduced as TILS-induced metabolic-hemodynamic coupling indices to quantify the coupling strength between TILS-enhanced cerebral metabolism and blood oxygen supply. This study provides the first demonstration that TILS causes up-regulation of oxidized CCO in the human brain, and contributes important insight into the physiological mechanisms.

  1. Development of tunable high pressure CO2 laser for lidar measurements of pollutants and wind velocities

    NASA Technical Reports Server (NTRS)

    Levine, J. S.; Guerra, M.; Javan, A.

    1980-01-01

    The problem of laser energy extraction at a tunable monochromatic frequency from an energetic high pressure CO2 pulsed laser plasma, for application to remote sensing of atmospheric pollutants by Differential Absorption Lidar (DIAL) and of wind velocities by Doppler Lidar, was investigated. The energy extraction principle analyzed is based on transient injection locking (TIL) at a tunable frequency. Several critical experiments for high gain power amplification by TIL are presented.

  2. Prognostic value, localization and correlation of PD-1/PD-L1, CD8 and FOXP3 with the desmoplastic stroma in pancreatic ductal adenocarcinoma

    PubMed Central

    Diana, Angela; Wang, Lai Mun; D'Costa, Zenobia; Allen, Paul; Azad, Abul; Silva, Michael A.; Soonawalla, Zahir; Liu, Stanley; McKenna, W. Gillies; Muschel, Ruth J.; Fokas, Emmanouil

    2016-01-01

    We examined the prognostic value of programmed cell death-1 (PD-1) and its ligand (PD-L1) together with CD8+ tumor-infiltrating lymphocytes (TILs) and FOXP3+ Tregs in resectable pancreatic ductal adenocarcinoma (PDAC) samples treated with adjuvant chemotherapy. Whole-mount FFPE tissue sections from 145 pancreatectomies were immunohistochemically stained for PD-1, PD-L1, CD8 and FOXP3. Their expression was correlated with clinicopathological characteristics, and overall survival (OS), progression-free survival (PFS), local progression-free survival (LPFS) and distant metastases free-survival (DMFS), in the context of stroma density (haematoxylin-eosin) and activity (alpha-smooth muscle actin) and in regard to intratumoral lymphoid aggregates. The median OS was 21 months after a mean follow-up of 20 months (range, 2-69 months). In multivariate analysis, high PD-1+ TILs expression was associated with better OS (p = 0.049), LPFS (p = 0.017) and DMFS (p = 0.021). Similar findings were observed for CD8+ TILs, whereas FOXP3 and PD-L1 lacked prognostic significance. Although TIL distribution was heterogeneous, tumors of high stroma density had higher infiltration of CD8+ TILs than loose density stroma and vice versa (p < 0.001), whereas no correlation was found with stromal activity. Sixty (41.4%) tumors contained lymphoid aggregates and the presence of PD-1+ TILs was associated with better OS (p = 0.030), LPFS (p = 0.025) and DMFS (p = 0.033), whereas CD8+ TILs only correlated with superior LPFS (p = 0.039). PD-1+ and CD8+ TILs constitute independent prognostic markers in patients with PDAC treated with adjuvant chemotherapy. Our study provides important insight on the role of PD-1/PD-L1 in the context of desmoplastic stroma and could help guide future immunotherapies in PDAC. PMID:27329602

  3. Identification by digital immunohistochemistry of intratumoral changes of immune infiltrates after vaccine in the absence of modifications of PBMC immune cell subsets.

    PubMed

    Farsaci, Benedetto; Jochems, Caroline; Grenga, Italia; Donahue, Renee N; Tucker, Jo A; Pinto, Peter A; Merino, Maria J; Heery, Christopher R; Madan, Ravi A; Gulley, James L; Schlom, Jeffrey

    2014-08-15

    Preclinical studies have demonstrated that the combination of systemic subcutaneous (s.c.) vaccination with intratumoral (i.t.) vaccination was superior in the induction of antitumor activity vs. vaccination with either route alone. A subsequent phase I study employing i.t.-s.c. vaccination was carried out in men with locally recurrent or progressive prostate cancer. rF-PSA-TRICOM (PROSTVAC) vaccine was administered intraprostatically and rV-PSA-TRICOM followed by rF-PSA-TRICOM vaccine was administered systemically. In that study no dose limiting toxicities were observed, 19/21 patients had stable or improved prostate-specific antigen (PSA) values and tumor-infiltrating lymphocytes (TILs) increased in post- vs. pre-treatment tumor biopsies, analyzed employing conventional immunohistochemistry (IHC). In the studies reported here, 31 phenotypes of peripheral blood mononuclear cells (PBMCs) were analyzed prevaccination and postvaccination as well as the functions of PBMC regulatory T cells (Tregs) and natural killer cells. A trend was observed in decreases in serum PSA with the reduction of circulating Tregs postvaccination. Digital IHC was employed prevaccination and postvaccination to measure CD4 and CD8 TILs, as well as Treg TILs by conventional IHC. Few correlations were observed with CD4, CD8 or Treg in TILs vs. PBMCs. However, patients with lower levels of CD4 TILs prevaccination showed the greatest increases in CD4 TILs postvaccine, while Treg TILs decreased postvaccine. There was also a strong correlation between decreases in serum PSA and increases in CD8 TILs postvaccine. These studies provide additional rationale for the use of i.t.-s.c. vaccinations and demonstrate a noncoordinate expression of specific immune subsets in PBMCs vs. tumor.

  4. Evaluation of the Toxicokinetic Study of 1,2-Propanediol Dinitrate (PGDN) in the Dog.

    DTIC Science & Technology

    1982-05-01

    JAFAMRL-TR-82-27 EVALUATION OF THE TOXICOKINETIC STUDY OF 1,2- PROPANEDIOL DINMITATE (PGDN) IN THE DOG P. K NEWTON J. A. MA CE WAN p- HV.R NOTm...Rltoo COVERED EVALUATION OF TIlE TOXICOKIXETIC STUDY OF 1,2- PROPANEDIOL DINITRATE (PGDN) IN TIlE Technical Report DOG 1 PERFORMNG OIG REPORT NUMAER...l,2- Propanediol Dinitrate ilypotension PGDN Tachycardia Toxicokinetics Dog *.ilethemoglobin Bolus injections of PGDN, an organic dinitrate, were made

  5. Targeting programmed cell death ligand 1 in osteosarcoma: an auto-commentary on therapeutic potential.

    PubMed

    Shen, Jacson K; Cote, Gregory M; Choy, Edwin; Hornicek, Francis J; Duan, Zhenfeng

    Programmed cell death ligand 1 (PDL1) expression was recently shown to correlate with tumor-infiltrating lymphocytes (TILs) in a subset of osteosarcoma patients. Among clinical factors evaluated across human osteosarcoma samples, a pulmonary origin of metastases correlated with high PDL1 expression and prominent TILs. Considering that multiple agents targeting PD-1/PDL1 are under development, targeting this immune checkpoint may be a novel immunotherapeutic route for osteosarcoma in future clinical trials.

  6. Tumor-infiltrating lymphocyte activity is enhanced in tumors with low IL-10 production in HBV-induced hepatocellular carcinoma

    SciTech Connect

    Shi, Yang Song, Qingwei; Hu, Dianhe; Zhuang, Xiaohu; Yu, Shengcai

    2015-05-22

    Hepatocellular carcinoma (HCC) is one of the most common cancers and can be induced by chronic HBV infection. The role of HBV-specific immune responses in mediating tumorigenesis and HCC prognosis is debated. The effect of intratumoral microenvironment on tumor-infiltrating lymphocytes (TILs) is also unclear. Here, we examined resected tumor tissue from 36 patients with HBV-induced HCC. We categorized study cohort based on ex vivo IL-10 secretion by tumor cells into high IL-10-secreting (Hi10) and low IL-10-secreting (Lo10) groups, and found that the Lo10 group was less sensitive to TLR ligand stimulation. TILs from the Lo10 group contained higher frequencies of HBV-specific IFN-g-producing cells and total IFN-g-producing cells, and possessed higher proliferative capacity. Moreover, the proliferative capacity of TILs from the Hi10 group was negatively correlated with IL-10 secretion from tumor cells. Together, our data demonstrated that low IL-10-producing capacity in HBV-induced HCC tumors is associated with enhanced TIL activity. - Highlights: • We examined intratumoral IL-10 production in HBV-induced HCC. • We grouped HCC tumors into Hi10 and Lo10 groups based on their IL-10 production. • Lo10 groups had better IFN-g response by TILs. • Lo10 groups had better TIL proliferative capacity. • Lo10 group tumor cells were refractory to TLR ligand stimulation.

  7. BTLA marks a less-differentiated tumor-infiltrating lymphocyte subset in melanoma with enhanced survival properties

    PubMed Central

    Haymaker, Cara L; Wu, Richard C; Ritthipichai, Krit; Bernatchez, Chantale; Forget, Marie-Andrée; Chen, Jie Qing; Liu, Hui; Wang, Ena; Marincola, Francesco; Hwu, Patrick; Radvanyi, Laszlo G

    2015-01-01

    In a recent adoptive cell therapy (ACT) clinical trial using autologous tumor-infiltrating lymphocytes (TILs) in patients with metastatic melanoma, we found an association between CD8+ T cells expressing the inhibitory receptor B- and T-lymphocyte attenuator (BTLA) and clinical response. Here, we further characterized this CD8+BTLA+ TIL subset and their CD8+BTLA− counterparts. We found that the CD8+ BTLA+ TILs had an increased response to IL-2, were less-differentiated effector-memory (TEM) cells, and persisted longer in vivo after infusion. In contrast, CD8+BTLA− TILs failed to proliferate and expressed genes associated with T-cell deletion/tolerance. Paradoxically, activation of BTLA signaling by its ligand, herpes virus entry mediator (HVEM), inhibited T-cell division and cytokine production, but also activated the Akt/PKB pathway thus protecting CD8+BTLA+ TILs from apoptosis. Our results point to a new role of BTLA as a useful T-cell differentiation marker in ACT and a dual signaling molecule that curtails T-cell activation while also conferring a survival advantage for CD8+ T cells. These attributes may explain our previous observation that BTLA expression on CD8+ TILs correlates with clinical response to adoptive T-cell therapy in metastatic melanoma. PMID:26405566

  8. Reprogramming of Melanoma Tumor-Infiltrating Lymphocytes to Induced Pluripotent Stem Cells

    PubMed Central

    Saito, Hidehito; Okita, Keisuke; Fusaki, Noemi; Sabel, Michael S.; Chang, Alfred E.; Ito, Fumito

    2016-01-01

    Induced pluripotent stem cells (iPSCs) derived from somatic cells of patients hold great promise for autologous cell therapies. One of the possible applications of iPSCs is to use them as a cell source for producing autologous lymphocytes for cell-based therapy against cancer. Tumor-infiltrating lymphocytes (TILs) that express programmed cell death protein-1 (PD-1) are tumor-reactive T cells, and adoptive cell therapy with autologous TILs has been found to achieve durable complete response in selected patients with metastatic melanoma. Here, we describe the derivation of human iPSCs from melanoma TILs expressing high level of PD-1 by Sendai virus-mediated transduction of the four transcription factors, OCT3/4, SOX2, KLF4, and c-MYC. TIL-derived iPSCs display embryonic stem cell-like morphology, have normal karyotype, express stem cell-specific surface antigens and pluripotency-associated transcription factors, and have the capacity to differentiate in vitro and in vivo. A wide variety of T cell receptor gene rearrangement patterns in TIL-derived iPSCs confirmed the heterogeneity of T cells infiltrating melanomas. The ability to reprogram TILs containing patient-specific tumor-reactive repertoire might allow the generation of patient- and tumor-specific polyclonal T cells for cancer immunotherapy. PMID:27057178

  9. Clinical Scale Zinc Finger Nuclease-mediated Gene Editing of PD-1 in Tumor Infiltrating Lymphocytes for the Treatment of Metastatic Melanoma

    PubMed Central

    Beane, Joal D; Lee, Gary; Zheng, Zhili; Mendel, Matthew; Abate-Daga, Daniel; Bharathan, Mini; Black, Mary; Gandhi, Nimisha; Yu, Zhiya; Chandran, Smita; Giedlin, Martin; Ando, Dale; Miller, Jeff; Paschon, David; Guschin, Dmitry; Rebar, Edward J; Reik, Andreas; Holmes, Michael C; Gregory, Philip D; Restifo, Nicholas P; Rosenberg, Steven A; Morgan, Richard A; Feldman, Steven A

    2015-01-01

    Programmed cell death-1 (PD-1) is expressed on activated T cells and represents an attractive target for gene-editing of tumor targeted T cells prior to adoptive cell transfer (ACT). We used zinc finger nucleases (ZFNs) directed against the gene encoding human PD-1 (PDCD-1) to gene-edit melanoma tumor infiltrating lymphocytes (TIL). We show that our clinical scale TIL production process yielded efficient modification of the PD-1 gene locus, with an average modification frequency of 74.8% (n = 3, range 69.9–84.1%) of the alleles in a bulk TIL population, which resulted in a 76% reduction in PD-1 surface-expression. Forty to 48% of PD-1 gene-edited cells had biallelic PD-1 modification. Importantly, the PD-1 gene-edited TIL product showed improved in vitro effector function and a significantly increased polyfunctional cytokine profile (TNFα, GM-CSF, and IFNγ) compared to unmodified TIL in two of the three donors tested. In addition, all donor cells displayed an effector memory phenotype and expanded approximately 500–2,000-fold in vitro. Thus, further study to determine the efficiency and safety of adoptive cell transfer using PD-1 gene-edited TIL for the treatment of metastatic melanoma is warranted. PMID:25939491

  10. Early T cell signalling is reversibly altered in PD-1+ T lymphocytes infiltrating human tumors.

    PubMed

    Wang, Shu-Fang; Fouquet, Stéphane; Chapon, Maxime; Salmon, Hélène; Regnier, Fabienne; Labroquère, Karine; Badoual, Cécile; Damotte, Diane; Validire, Pierre; Maubec, Eve; Delongchamps, Nicolas B; Cazes, Aurélie; Gibault, Laure; Garcette, Marylène; Dieu-Nosjean, Marie-Caroline; Zerbib, Marc; Avril, Marie-Françoise; Prévost-Blondel, Armelle; Randriamampita, Clotilde; Trautmann, Alain; Bercovici, Nadège

    2011-03-07

    To improve cancer immunotherapy, a better understanding of the weak efficiency of tumor-infiltrating T lymphocytes (TIL) is necessary. We have analyzed the functional state of human TIL immediately after resection of three types of tumors (NSCLC, melanoma and RCC). Several signalling pathways (calcium, phosphorylation of ERK and Akt) and cytokine secretion are affected to different extents in TIL, and show a partial spontaneous recovery within a few hours in culture. The global result is an anergy that is quite distinct from clonal anergy induced in vitro, and closer to adaptive tolerance in mice. PD-1 (programmed death -1) is systematically expressed by TIL and may contribute to their anergy by its mere expression, and not only when it interacts with its ligands PD-L1 or PD-L2, which are not expressed by every tumor. Indeed, the TCR-induced calcium and ERK responses were reduced in peripheral blood T cells transfected with PD-1. Inhibition by sodium stibogluconate of the SHP-1 and SHP-2 phosphatases that associate with several inhibitory receptors including PD-1, relieves part of the anergy apparent in TIL or in PD-1-transfected T cells. This work highlights some of the molecular modifications contributing to functional defects of human TIL.

  11. Clinical Scale Zinc Finger Nuclease-mediated Gene Editing of PD-1 in Tumor Infiltrating Lymphocytes for the Treatment of Metastatic Melanoma.

    PubMed

    Beane, Joal D; Lee, Gary; Zheng, Zhili; Mendel, Matthew; Abate-Daga, Daniel; Bharathan, Mini; Black, Mary; Gandhi, Nimisha; Yu, Zhiya; Chandran, Smita; Giedlin, Martin; Ando, Dale; Miller, Jeff; Paschon, David; Guschin, Dmitry; Rebar, Edward J; Reik, Andreas; Holmes, Michael C; Gregory, Philip D; Restifo, Nicholas P; Rosenberg, Steven A; Morgan, Richard A; Feldman, Steven A

    2015-08-01

    Programmed cell death-1 (PD-1) is expressed on activated T cells and represents an attractive target for gene-editing of tumor targeted T cells prior to adoptive cell transfer (ACT). We used zinc finger nucleases (ZFNs) directed against the gene encoding human PD-1 (PDCD-1) to gene-edit melanoma tumor infiltrating lymphocytes (TIL). We show that our clinical scale TIL production process yielded efficient modification of the PD-1 gene locus, with an average modification frequency of 74.8% (n = 3, range 69.9-84.1%) of the alleles in a bulk TIL population, which resulted in a 76% reduction in PD-1 surface-expression. Forty to 48% of PD-1 gene-edited cells had biallelic PD-1 modification. Importantly, the PD-1 gene-edited TIL product showed improved in vitro effector function and a significantly increased polyfunctional cytokine profile (TNFα, GM-CSF, and IFNγ) compared to unmodified TIL in two of the three donors tested. In addition, all donor cells displayed an effector memory phenotype and expanded approximately 500-2,000-fold in vitro. Thus, further study to determine the efficiency and safety of adoptive cell transfer using PD-1 gene-edited TIL for the treatment of metastatic melanoma is warranted.

  12. Sunitinib pretreatment improves tumor-infiltrating lymphocyte expansion by reduction in intratumoral content of myeloid-derived suppressor cells in human renal cell carcinoma.

    PubMed

    Guislain, Aurelie; Gadiot, Jules; Kaiser, Andrew; Jordanova, Ekaterina S; Broeks, Annegien; Sanders, Joyce; van Boven, Hester; de Gruijl, Tanja D; Haanen, John B A G; Bex, Axel; Blank, Christian U

    2015-10-01

    Targeted therapy with sunitinib, pazopanib or everolimus has improved treatment outcome for patients with metastatic renal cell carcinoma patients (RCC). However, despite considerable efforts in sequential or combined modalities, durable remissions are rare. Immunotherapy like cytokine therapy with interleukin-2, T cell checkpoint blockade or adoptive T cell therapies can achieve long-term benefit and even cure. This raises the question of whether combining targeted therapy with immunotherapy could also be an effective treatment option for RCC patients. Sunitinib, one of the most frequently administered therapeutics in RCC patients has been implicated in impairing T cell activation and proliferation in vitro. In this work, we addressed whether this notion holds true for expansion of tumor-infiltrating lymphocytes (TILs) in sunitinib-treated patients. We compared resected primary RCC tumor material of patients pretreated with sunitinib with resection specimen from sunitinib-naïve patients. We found improved TIL expansion from sunitinib-pretreated tumor digests. These TIL products contained more PD-1 expressing TIL, while the regulatory T cell infiltration was not altered. The improved TIL expansion was associated with reduced intratumoral myeloid-derived suppressor cell (MDSC) content. Depletion of MDSCs from sunitinib-naïve RCC tissue-digest improved TIL expansion, proving the functional relevance of the MDSC alteration by sunitinib. Our in vivo results do not support previous in vitro observations of sunitinib inhibiting T cell function, but do provide a possible rationale for the combination of sunitinib with immunotherapy.

  13. Mitochondrial DNA toxicity in forebrain neurons causes apoptosis, neurodegeneration, and impaired behavior.

    PubMed

    Lauritzen, Knut H; Moldestad, Olve; Eide, Lars; Carlsen, Harald; Nesse, Gaute; Storm, Johan F; Mansuy, Isabelle M; Bergersen, Linda H; Klungland, Arne

    2010-03-01

    Mitochondrial dysfunction underlying changes in neurodegenerative diseases is often associated with apoptosis and a progressive loss of neurons, and damage to the mitochondrial genome is proposed to be involved in such pathologies. In the present study we designed a mouse model that allows us to specifically induce mitochondrial DNA toxicity in the forebrain neurons of adult mice. This is achieved by CaMKIIalpha-regulated inducible expression of a mutated version of the mitochondrial UNG DNA repair enzyme (mutUNG1). This enzyme is capable of removing thymine from the mitochondrial genome. We demonstrate that a continual generation of apyrimidinic sites causes apoptosis and neuronal death. These defects are associated with behavioral alterations characterized by increased locomotor activity, impaired cognitive abilities, and lack of anxietylike responses. In summary, whereas mitochondrial base substitution and deletions previously have been shown to correlate with premature and natural aging, respectively, we show that a high level of apyrimidinic sites lead to mitochondrial DNA cytotoxicity, which causes apoptosis, followed by neurodegeneration.

  14. A ketogenic diet accelerates neurodegeneration in mice with induced mitochondrial DNA toxicity in the forebrain.

    PubMed

    Lauritzen, Knut H; Hasan-Olive, Md Mahdi; Regnell, Christine E; Kleppa, Liv; Scheibye-Knudsen, Morten; Gjedde, Albert; Klungland, Arne; Bohr, Vilhelm A; Storm-Mathisen, Jon; Bergersen, Linda H

    2016-12-01

    Mitochondrial genome maintenance plays a central role in preserving brain health. We previously demonstrated accumulation of mitochondrial DNA damage and severe neurodegeneration in transgenic mice inducibly expressing a mutated mitochondrial DNA repair enzyme (mutUNG1) selectively in forebrain neurons. Here, we examine whether severe neurodegeneration in mutUNG1-expressing mice could be rescued by feeding the mice a ketogenic diet, which is known to have beneficial effects in several neurological disorders. The diet increased the levels of superoxide dismutase 2, and mitochondrial mass, enzymes, and regulators such as SIRT1 and FIS1, and appeared to downregulate N-methyl-D-aspartic acid (NMDA) receptor subunits NR2A/B and upregulate γ-aminobutyric acid A (GABAA) receptor subunits α1. However, unexpectedly, the ketogenic diet aggravated neurodegeneration and mitochondrial deterioration. Electron microscopy showed structurally impaired mitochondria accumulating in neuronal perikarya. We propose that aggravation is caused by increased mitochondrial biogenesis of generally dysfunctional mitochondria. This study thereby questions the dogma that a ketogenic diet is unambiguously beneficial in mitochondrial disorders.

  15. The Alfred Nobel rocket camera. An early aerial photography attempt

    NASA Astrophysics Data System (ADS)

    Ingemar Skoog, A.

    2010-02-01

    Alfred Nobel (1833-1896), mainly known for his invention of dynamite and the creation of the Nobel Prices, was an engineer and inventor active in many fields of science and engineering, e.g. chemistry, medicine, mechanics, metallurgy, optics, armoury and rocketry. Amongst his inventions in rocketry was the smokeless solid propellant ballistite (i.e. cordite) patented for the first time in 1887. As a very wealthy person he actively supported many Swedish inventors in their work. One of them was W.T. Unge, who was devoted to the development of rockets and their applications. Nobel and Unge had several rocket patents together and also jointly worked on various rocket applications. In mid-1896 Nobel applied for patents in England and France for "An Improved Mode of Obtaining Photographic Maps and Earth or Ground Measurements" using a photographic camera carried by a "…balloon, rocket or missile…". During the remaining of 1896 the mechanical design of the camera mechanism was pursued and cameras manufactured. In April 1897 (after the death of Alfred Nobel) the first aerial photos were taken by these cameras. These photos might be the first documented aerial photos taken by a rocket borne camera. Cameras and photos from 1897 have been preserved. Nobel did not only develop the rocket borne camera but also proposed methods on how to use the photographs taken for ground measurements and preparing maps.

  16. Interaktive Visualisierung von Abständen und Ausdehnungen anatomischer Strukturen für die Interventionsplanung

    NASA Astrophysics Data System (ADS)

    Rössling, Ivo; Cyrus, Christian; Dornheim, Lars; Hahn, Peter; Preim, Bernhard; Boehm, Andreas

    Im Rahmen der Interventionsplanung muss der Chirurg therapierelevante Entscheidungen auf Basis räumlicher Relationen anatomischer Strukturen treffen. Interaktive 3D-Visualisierungen unterstützen diesen Prozess qualitativ. Quantitative Fragestellungen (Tumorausdehnung, Infiltrationstiefe, etc.) erfordern die Integration einer Bemaßung, deren Nutzen wesentlich von einer geeigneten Darstellung abhängt. In dieser Arbeit haben wir allgemeine Kriterien für die Eignung von Visualisierungen von Bemaßungen in interaktiven 3D-Szenen erarbeitet. Daran orientierend haben wir verschiedene Varianten der Darstellung von Abständen und Ausdehnungen anatomischer Strukturen betrachtet und ihr Erscheinungsbild hierzu zweckmäßig parametrisiert. Die Ausprägungen dieser Darstellungsparameter wurden in einer Studie auf ihre visuellen Wirkung hin an Chirurgen evaluiert. Es zeigte sich, dass die befragten Mediziner höchsten Wert auf Kohärenz und klare Zuordnung der Bemaßung setzten und überraschenderweise dafür sogar Abstriche in der direkten Lesbarkeit in Kauf nahmen.

  17. Non-canonical uracil processing in DNA gives rise to double-strand breaks and deletions: relevance to class switch recombination

    PubMed Central

    Bregenhorn, Stephanie; Kallenberger, Lia; Artola-Borán, Mariela; Peña-Diaz, Javier; Jiricny, Josef

    2016-01-01

    During class switch recombination (CSR), antigen-stimulated B-cells rearrange their immunoglobulin constant heavy chain (CH) loci to generate antibodies with different effector functions. CSR is initiated by activation-induced deaminase (AID), which converts cytosines in switch (S) regions, repetitive sequences flanking the CH loci, to uracils. Although U/G mispairs arising in this way are generally efficiently repaired to C/Gs by uracil DNA glycosylase (UNG)-initiated base excision repair (BER), uracil processing in S-regions of activated B-cells occasionally gives rise to double strand breaks (DSBs), which trigger CSR. Surprisingly, genetic experiments revealed that CSR is dependent not only on AID and UNG, but also on mismatch repair (MMR). To elucidate the role of MMR in CSR, we studied the processing of uracil-containing DNA substrates in extracts of MMR-proficient and –deficient human cells, as well as in a system reconstituted from recombinant BER and MMR proteins. Here, we show that the interplay of these repair systems gives rise to DSBs in vitro and to genomic deletions and mutations in vivo, particularly in an S-region sequence. Our findings further suggest that MMR affects pathway choice in DSB repair. Given its amenability to manipulation, our system represents a powerful tool for the molecular dissection of CSR. PMID:26743004

  18. Surface Proteome Analysis of a Natural Isolate of Lactococcus lactis Reveals the Presence of Pili Able to Bind Human Intestinal Epithelial Cells*

    PubMed Central

    Meyrand, Mickael; Guillot, Alain; Goin, Mélodie; Furlan, Sylviane; Armalyte, Julija; Kulakauskas, Saulius; Cortes-Perez, Naima G.; Thomas, Ginette; Chat, Sophie; Péchoux, Christine; Dupres, Vincent; Hols, Pascal; Dufrêne, Yves F.; Trugnan, Germain; Chapot-Chartier, Marie-Pierre

    2013-01-01

    Surface proteins of Gram-positive bacteria play crucial roles in bacterial adhesion to host tissues. Regarding commensal or probiotic bacteria, adhesion to intestinal mucosa may promote their persistence in the gastro-intestinal tract and their beneficial effects to the host. In this study, seven Lactococcus lactis strains exhibiting variable surface physico-chemical properties were compared for their adhesion to Caco-2 intestinal epithelial cells. In this test, only one vegetal isolate TIL448 expressed a high-adhesion phenotype. A nonadhesive derivative was obtained by plasmid curing from TIL448, indicating that the adhesion determinants were plasmid-encoded. Surface-exposed proteins in TIL448 were analyzed by a proteomic approach consisting in shaving of the bacterial surface with trypsin and analysis of the released peptides by LC-MS/MS. As the TIL448 complete genome sequence was not available, the tryptic peptides were identified by a mass matching approach against a database including all Lactococcus protein sequences and the sequences deduced from partial DNA sequences of the TIL448 plasmids. Two surface proteins, encoded by plasmids in TIL448, were identified as candidate adhesins, the first one displaying pilin characteristics and the second one containing two mucus-binding domains. Inactivation of the pilin gene abolished adhesion to Caco-2 cells whereas inactivation of the mucus-binding protein gene had no effect on adhesion. The pilin gene is located inside a cluster of four genes encoding two other pilin-like proteins and one class-C sortase. Synthesis of pili was confirmed by immunoblotting detection of high molecular weight forms of pilins associated to the cell wall as well as by electron and atomic force microscopy observations. As a conclusion, surface proteome analysis allowed us to detect pilins at the surface of L. lactis TIL448. Moreover we showed that pili appendages are formed and involved in adhesion to Caco-2 intestinal epithelial cells

  19. Tumour-infiltrating lymphocytes and the emerging role of immunotherapy in breast cancer.

    PubMed

    Luen, Stephen J; Savas, Peter; Fox, Stephen B; Salgado, Roberto; Loi, Sherene

    2017-02-01

    Breast cancer has not previously been considered a highly immunogenic cancer. Observations of tumour-infiltrating lymphocytes (TILs) in and around neoplastic cells in tumour samples, and associations with improved pathological complete response and clinical survival end points have changed our perspective on this. Lymphocytic infiltrates have long been observed in breast cancer; however, more recently, retrospective analysis of prospectively collected tissue samples from clinical trials has demonstrated the potential role of host immunosurveillance in influencing the biology of breast cancer. This association appears to be strongest in triple negative and HER2 positive breast cancer subtypes. Contrastingly, the association in luminal tumours is less clear, and is potentially limited by substantial tumoural heterogeneity. Several methodologies have been employed to quantify, and describe the composition of TILs, each with its own advantages and disadvantages. The results of these analyses have been generally consistent, and valuable efforts are currently underway to standardise the evaluation of TILs toward a universal approach. More technical methods of TIL characterisation remain important in the research setting. The evaluation of TILs becomes increasingly relevant with the emerging role of immunotherapy in breast cancer. Early phase trials of checkpoint blockade show promising results; however, it is likely that some patients will require combination treatments to maximise therapeutic benefits. Equally, some patients may not derive any benefit from immunotherapies. This underscores the importance of the development of relevant predictive biomarkers. As a key representative of the immune interaction between host and tumour, lymphocytic infiltrates are ideally placed for continued research into the determinants of immunogenicity, and response to immunotherapeutic approaches. In this review, we will discuss the current methodologies of evaluation, and the clinical

  20. Correlation between class I antigen expression and the ability to generate tumour infiltrating lymphocytes from bladder tumour biopsies.

    PubMed Central

    Nouri, A. M.; dos Santos, A. V.; Crosby, D.; Oliver, R. T.

    1991-01-01

    Analysis of tissue sections from transurethrally resected bladder tumours using anti-CD3 antibody showed the presence of T lymphocytes in intra-epithelial layers in eight of 12 cases investigated. In a larger group of patients, Tumour Infiltrating Lymphocyte (TIL) growth was established from six of 19 cases using Interleukin-2 (IL-2) and conditioned medium (CM) and resulted in the expansion of TILs up to 100-fold. TILs from these individuals were phenotyped with W6/32 (anti-HLA-A,B,C), HB55 (anti-DR) and anti-CD3 antibodies using FAC sorter. The mean +/- s.d. frequency of positive staining with these antibodies were 96.7 +/- 4.0%, 87.5 +/- 10.0% and 82.5 +/- 7.8% respectively, indicating the activated nature of these T cells. The cytotoxic activity of these TILs against Daudi (ie, LAK activity) cell line at 25/1 E/T ratios varied from 26.3 +/- 3.2 to 62.8 +/- 5.2%. In one case where TILs and autologous tumour cell line were established, cytotoxicity studies showed low level of cytotoxicity against the autologous tumour cells (15.8 +/- 1.6%) compared with 62.8 +/- 5.2% against Daudi. Staining of tumour sections from these 19 individuals with W6/32 and BBM.1 revealed positive staining in six of six that developed TILs but only six of 13 (46%) cases, whose tumour failed to grow TILs (P less than 0.02, Fisher exact test). These results are indicative of the presence of IL-2 passageable T cells in bladder cancer biopsy and demonstrate that the successful expansion of these cells correlates with the normal expression of class I antigens on the tumour cells. Images Figure 5 PMID:1764393

  1. Neoadjuvant Chemotherapy of Ovarian Cancer Results in Three Patterns of Tumor-Infiltrating Lymphocyte Response with Distinct Implications for Immunotherapy.

    PubMed

    Lo, Charlotte S; Sanii, Sanaz; Kroeger, David R; Milne, Katy; Talhouk, Aline; Chiu, Derek S; Rahimi, Kurosh; Shaw, Patricia A; Clarke, Blaise A; Nelson, Brad H

    2017-02-15

    Purpose: Some forms of chemotherapy can enhance antitumor immunity through immunogenic cell death, resulting in increased T-cell activation and tumor infiltration. Such effects could potentially sensitize tumors to immunotherapies, including checkpoint blockade. We investigated whether platinum- and taxane-based chemotherapy for ovarian cancer induces immunologic changes consistent with this possibility.Experimental Design: Matched pre- and post-neoadjuvant chemotherapy tumor samples from 26 high-grade serous carcinoma (HGSC) patients were analyzed by immunohistochemistry (IHC) for a large panel of immune cells and associated factors. The prognostic significance of post-chemotherapy TIL patterns was assessed in an expanded cohort (n = 90).Results: Neoadjuvant chemotherapy was associated with increased densities of CD3(+), CD8(+), CD8(+) TIA-1(+), PD-1(+) and CD20(+) TIL. Other immune subsets and factors were unchanged, including CD79a(+) CD138(+) plasma cells, CD68(+) macrophages, and MHC class I on tumor cells. Immunosuppressive cell types were also unchanged, including FoxP3(+) PD-1(+) cells (putative regulatory T cells), IDO-1(+) cells, and PD-L1(+) cells (both macrophages and tumor cells). Hierarchical clustering revealed three response patterns: (i) TIL(high) tumors showed increases in multiple immune markers after chemotherapy; (ii) TIL(low) tumors underwent similar increases, achieving patterns indistinguishable from the first group; and (iii) TIL(negative) cases generally remained negative. Despite the dramatic increases seen in the first two patterns, post-chemotherapy TIL showed limited prognostic significance.Conclusions: Chemotherapy augments pre-existing TIL responses but fails to relieve major immune-suppressive mechanisms or confer significant prognostic benefit. Our findings provide rationale for multipronged approaches to immunotherapy tailored to the baseline features of the tumor microenvironment. Clin Cancer Res; 23(4); 925-34. ©2016 AACR.

  2. Numerical studies on the performance of an aerosol respirator with faceseal leakage

    NASA Astrophysics Data System (ADS)

    Zaripov, S. K.; Mukhametzanov, I. T.; Grinshpun, S. A.

    2016-11-01

    We studied the efficiency of a facepiece filtering respirator (FFR) in presence of a measurable faceseal leakage using the previously developed model of a spherical sampler with porous layer. In our earlier study, the model was validated for a specific filter permeability value. In this follow-up study, we investigated the effect of permeability on the overall respirator performance accounting for the faceseal leakage. The Total Inward Leakage (TIL) was calculated as a function of the leakage-to-filter surface ratio and the particle diameter. A good correlation was found between the theoretical and experimental TIL values. The TIL value was shown to increase and the effect of particle size on TIL to decrease as the leakage-to- filter surface ratio grows. The model confirmed that within the most penetrating particle size range (∼50 nm) and at relatively low leakage-to-filter surface ratios, an FFR performs better (TIL is lower) when the filter has a lower permeability which should be anticipated as long as the flow through the filter represents the dominant particle penetration pathway. An increase in leak size causes the TIL to rise; furthermore, under certain leakage-to-filter surface ratios, TIL for ultrafine particles becomes essentially independent on the filter properties due to a greater contribution of the aerosol flow through the faceseal leakage. In contrast to the ultrafine fraction, the larger particles (e.g., 800 nm) entering a typical high- or medium-quality respirator filter are almost fully collected by the filter medium regardless of its permeability; at the same time, the fraction penetrated through the leakage appears to be permeability- dependent: higher permeability generally results in a lower pressure drop through the filter which increases the air flow through the filter at the expense of the leakage flow. The latter reduces the leakage effect thus improving the overall respiratory protection level. The findings of this study provide

  3. Tumor infiltrating T lymphocytes expressing FoxP3, CCR7 or PD-1 predict the outcome of prostate cancer patients subjected to salvage radiotherapy after biochemical relapse.

    PubMed

    Nardone, Valerio; Botta, Cirino; Caraglia, Michele; Martino, Elodia Claudia; Ambrosio, Maria Raffaella; Carfagno, Tommaso; Tini, Paolo; Semeraro, Leonardo; Misso, Gabriella; Grimaldi, Anna; Boccellino, Mariarosaria; Facchini, Gaetano; Berretta, Massimiliano; Vischi, Gianluca; Rocca, Bruno Jim; Barone, Aurora; Tassone, Pierfrancesco; Tagliaferri, Pierosandro; Del Vecchio, Maria Teresa; Pirtoli, Luigi; Correale, Pierpaolo

    2016-11-01

    Tumor immunologic microenvironment is strongly involved in tumor progression and the presence of tumor infiltrating lymphocytes (TIL) with different phenotypes has been demonstrated to be of prognostic relevance in different malignancies. We investigated whether TIL infiltration of tumor tissues could also predict the outcome of prostate cancer patients. To this end, we carried out a retrospective analysis correlating the outcome of locally advanced prostate cancer patients undergone salvage radiotherapy upon relapse after radical surgery with the infiltration by different TIL populations. Twenty-two patients with resectable prostate cancer, with a mean age of 67 (+/-3.93) years, who received salvage radiotherapy with a mean of 69.66 (+/- 3.178) Gy in 8 weeks, between June 1999 and January 2009 and with a median follow up of 123 (+/- 55.82) months, were enrolled in this study. We evaluated, by immunohistochemistry, the intratumoral ((t)) and peripheral stroma ((p)) infiltration by CD45, CD3, CD4, CD8, CCR7, FoxP3 or PD-1-positive cells on tumor samples taken at the diagnosis ((d)) and relapse times ((R)). We correlated these variables with patients' biochemical progression free survival (bPFS), post-radiotherapy progression free survival (PFS), and overall survival (OS). Substantial changes in the rate of TIL subsets were found between the first and the second biopsy with progressive increase in CD4, CCR7, FoxP3, PD-1(+) cells. Our analysis revealed that higher CD8(p,R+) and lower PD-1(R+) TIL scores correlated to a longer bPFS. Higher CD8(p,R+) and CCR7(t,R+) TIL scores and lower CD45(p,R+) and FoxP3(p,R+) TIL scores correlated to a prolonged PFS and OS. These results suggest that the immunological microenvironment of primary tumor is strictly correlated with patient outcome and provide the rationale for immunological treatment of prostate cancer.

  4. Prognostic and predictive value of tumor-infiltrating lymphocytes for clinical therapeutic research in patients with non-small cell lung cancer

    PubMed Central

    Zeng, Dong-Qiang; Yu, Yun-Fang; Ou, Qi-Yun; Li, Xiao-Yin; Zhong, Ru-Zhi; Xie, Chuan-Miao; Hu, Qiu-Gen

    2016-01-01

    Background Previous preclinical and clinical studies have shown that levels of tumor-infiltrating lymphocytes (TILs) significantly correlated with prognosis in non-small cell lung cancer (NSCLC), and survival after therapy; however, this finding remains controversial. We performed a meta-analysis, to evaluate, systematically, the clinical utilization of TIL subtypes in patients with NSCLC. Methods The PubMed, ISI Web of Science, EMBASE, and Cochrane Library databases were searched to identify relevant studies. We pooled estimates of treatment effects, and hazards were summarized using random or fixed effects models to evaluate survival outcomes. Results A total of 24 relevant studies involving 7,006 patients were eligible. The median percentage of lymph node positivity was 45.7% (95% confidence interval [CI], 37.1–56.4%). Pooled analysis shows that high levels of CD8+ TILs had a good prognostic effect on survival with a hazard ratio (HR) of 0.91 (P = 0.013) for death and 0.74 (P = 0.001) for recurrence, as did high levels of CD3+ and CD4+ TILs, with HRs of 0.77 (P = 0.009) and 0.78 (P = 0.005) for death, respectively. By contrast, high levels of FoxP3+ regulatory TILs had a worse prognostic effect for overall and recurrence-free survival, with HRs of 1.69 (P = 0.042) and 1.79 (P = 0.001), respectively. No individual study affected the results, and no publication bias was found. Conclusions Our findings support the hypothesis that TILs could be a prognostic marker in NSCLC. High-quality randomized studies are needed to verify statistically the effect of TILs on prognosis in future research. PMID:26871598

  5. Identification and Functional Analysis of Interleukin-1β in the Chinese Soft-Shelled Turtle Pelodiscus sinensis

    PubMed Central

    Liang, Quan; Li, Weifen; Guo, Ningning; Tong, Chao; Zhou, Yingshan; Fang, Weihuan; Li, Xiaoliang

    2016-01-01

    Chinese soft-shelled turtle (Pelodiscus sinensis) is commercially cultured in East and Southeast Asia for its nutritional and medicinal values. In this study, we identified interleukin-1β (IL-1β) from Chinese soft-shelled turtle. The full-length cDNA of Pelodiscus sinensis IL-1β (tIL-1β) consists of 1529 base pairs with an 831-base-pair open reading frame, encoding 277 amino acids. The guanine-cytosine (GC) content in the coding sequence and 3’ untranslated region of tIL-1β is considerably higher than that of other vertebrates. Its mRNA expression level increased significantly during Aeromonas hydrophila infection. The tIL-1β lacks the typical IL-1β-converting enzyme (ICE) cut site found in mammalian IL-1β, but still could be cleaved by turtle caspase-1. By mutating the potential cleavage sites, we identified aspartic acid (Asp/D) 130 as the ICE cut site in tIL-1β. The peptide truncated at D130 was expressed using the baculovirus expression system; its bioactivity is confirmed by the ability to induce cyclooxygenase-2 (COX-2) and tIL-1β itself in peripheral blood monocytes. In conclusion, we characterized IL-1β from Chinese soft-shelled turtle and identified its D130 as a non-typical ICE cut size. PMID:27153094

  6. CD8+ tumor-infiltrating lymphocytes predict favorable prognosis in malignant pleural mesothelioma after resection.

    PubMed

    Yamada, Noriyuki; Oizumi, Satoshi; Kikuchi, Eiki; Shinagawa, Naofumi; Konishi-Sakakibara, Jun; Ishimine, Atsushi; Aoe, Keisuke; Gemba, Kenichi; Kishimoto, Takumi; Torigoe, Toshihiko; Nishimura, Masaharu

    2010-10-01

    Defects in human leukocyte antigen (HLA) class I expression may allow tumor cells to escape immune recognition. T cell infiltration is associated with a good prognosis in many cancers. However, the role of HLA class I expression and tumor-infiltrating lymphocytes (TILs) in malignant pleural mesothelioma (MPM) has not been fully analyzed. In the present study, we investigated the immune profiles and conducted outcome analyses of MPM patients. HLA class I expression and TILs (CD4(+), CD8(+), and NK cells) were detected by immunohistochemistry in a series of 44 MPM cases. To detect HLA class I expression, specimens were stained with the anti-pan HLA class I monoclonal antibody EMR8-5. The expression of HLA class I was positive in all patients. There was no case that showed negative HLA class I expression. The density of CD4(+) and CD8(+) TILs were strongly correlated (R = 0.76, p < 0.001). A high density of CD8(+) TILs was a significantly better prognostic factor for the survival of patients with extrapleural pneumonectomy (p < 0.05). Multivariate analysis revealed that a high density of CD8(+) TILs is an independent prognostic factor for patients who underwent extrapleural pneumonectomy. The presence of intratumoral CD8(+) T cells was correlated with an improved clinical outcome, raising the possibility that CD8(+) T cells might play a pivotal role in the antitumor immune response against MPMs. Thus, the stimulation of CD8(+) lymphocytes might be an efficacious immunotherapy for MPM patients.

  7. Tumor-Infiltrating Lymphocytes in Triple Negative Breast Cancer: The Future of Immune Targeting.

    PubMed

    García-Teijido, Paula; Cabal, María Luque; Fernández, Ignacio Peláez; Pérez, Yolanda Fernández

    2016-01-01

    Triple negative breast cancer (TNBC) is a highly heterogeneous tumor. There is increasing evidence of the role of tumor lymphocytic immune infiltrates in this subtype of breast cancer. Robust levels of tumor infiltrating lymphocytes (TILs) have been associated with improved disease-free and overall survival rates in TNBC patients with and without any treatment. Recent efforts have been made to develop a standardized methodology for evaluating TILs. The presence of TILs in the breast tumor microenvironment can also predict responses not only to neoadjuvant but also to adjuvant chemotherapy treatments. High numbers of TILs correlate with increased pathological complete responses (pCR) in TNBC. TILs are prognostic and predictive of response to standard therapies; thus, the immune system appears to play an active role in a subgroup of breast cancer. There is an increasing interest in directly targeting the immune system as part of breast cancer therapy, mainly in patients with TNBC. New immune modulatory agents, including immune checkpoints inhibitors, have shown promising activity in a subgroup of metastatic TNBC. Increased programmed cell death protein 1 ligand (PD-L1) expression on the surface of TNBC provides the rationale for implementing therapeutic strategies targeting the PD-1/PD-L1 axis in TNBC. The programmed cell death protein 1 (PD-1) inhibitor pembrolizumab, and the PD-L1 inhibitor atezolizumab have shown promising results in clinical trials.

  8. A major secretory defect of tumour-infiltrating T lymphocytes due to galectin impairing LFA-1-mediated synapse completion

    PubMed Central

    Petit, Anne-Elisabeth; Demotte, Nathalie; Scheid, Benoît; Wildmann, Claude; Bigirimana, René; Gordon-Alonso, Monica; Carrasco, Javier; Valitutti, Salvatore; Godelaine, Danièle; van der Bruggen, Pierre

    2016-01-01

    Surface galectin has been shown to contribute to dysfunctions of human tumour-infiltrating lymphocytes (TILs). We show here that galectin-covered CD8 TILs produce normal amounts of intracellular cytokines, but fail to secrete them because of defective actin rearrangements at the synapse. The non-secreting TILs also display reduced adhesion to their targets, together with defective LFA-1 recruitment and activation at the synapse. These defects are relieved by releasing surface galectin. As mild LFA-1 blockade on normal blood T cells emulate the defects of galectin-covered TILs, we conclude that galectin prevents the formation of a functional secretory synapse by preventing optimal LFA-1 triggering. Our results highlight a major secretory defect of TILs that is not revealed by widely used intracellular cytokine immunomonitoring assays. They also provide additional insights into the T-cell response, by showing that different thresholds of LFA-1 triggering are required to promote the intracellular production of cytokines and their secretion. PMID:27447355

  9. CD103+ intraepithelial T cells in high-grade serous ovarian cancer are phenotypically diverse TCRαβ+ CD8αβ+ T cells that can be targeted for cancer immunotherapy

    PubMed Central

    Workel, Hagma H.; Tijans, Aline M.; Terwindt, Anouk L.J.; Brunekreeft, Kim L.; Plat, Annechien; Klip, Harry G.; Eggink, Florine A.; Leffers, Ninke; Helfrich, Wijnand; Samplonius, Douwe F.; Bremer, Edwin; Wisman, G. Bea A.; Daemen, Toos; Duiker, Evelien W.; Hollema, Harry; Nijman, Hans W.; de Bruyn, Marco

    2016-01-01

    CD103+ tumor-infiltrating lymphocytes (TIL) have been linked to specific epithelial infiltration and a prolonged survival in high-grade serous epithelial ovarian cancer (HGSC). However, whether these cells are induced as part of an ongoing anti-HGSC immune response or represent non-specifically expanded resident or mucosal lymphocytes remains largely unknown. In this study, we first confirmed that CD103+ TIL from HGSC were predominantly localized in the cancer epithelium and were strongly correlated with an improved prognosis. We further demonstrate that CD103+ TIL were almost exclusively CD3+ TCRαβ+ CD8αβ+ CD4- T cells, but heterogeneously expressed T cell memory and differentiation markers. Activation of peripheral T cells in the presence of HGSC was sufficient to trigger induction of CD103 in over 90% of all CD8+ cells in a T cell receptor (TCR)- and TGFβR1-dependent manner. Finally, CD103+ TIL isolated from primary HGSC showed signs of recent activation and dominantly co-expressed key immunotherapeutic targets PD-1 and CD27. Taken together, our data indicate CD103+ TIL in HGSC are formed as the result of an adaptive anti-tumor immune response that might be reactivated by (dual) checkpoint inhibition. PMID:27650547

  10. Ex-vivo analysis of CD8+ T cells infiltrating colorectal tumors identifies a major effector-memory subset with low perforin content.

    PubMed

    Ye, Sheng-Wei; Wang, Yu; Valmori, Danila; Ayyoub, Maha; Han, Yan; Xu, Xiao-Lan; Zhao, Ai-Lian; Qu, Li; Gnjatic, Sacha; Ritter, Gerd; Old, Lloyd J; Gu, Jin

    2006-09-01

    Previous studies have indicated that the infiltration of CD8+ T cells in colorectal cancer is an independent predictor of increased survival but clinical observations have suggested that the cytotoxic function of CD8+ T cells infiltrating colorectal cancer may often be limited. In this study, we have assessed the phenotype of colorectal cancer CD8+ tumor-infiltrating lymphocytes (TILs) isolated ex vivo from tumor tissue, and assessed the perforin content of TIL with respect to their location using immunohistochemistry. We found that CD8+ T cells TILs isolated from colorectal cancer are mainly composed of antigen-experienced cells of effector memory type (TEM, CD45RA-CCR7-, and CD27+/CD28- or CD27-/CD28-), and contain only minor proportions of terminally differentiated CD8+ T cells (TEMRA, CD45RA+CCR7-). The perforin content of these TILs, however, is significantly lower than that of antigen-experienced T cells in PBMCs due to the much lower levels of perforin found in the CD27-CD28- subset in TILs compared with CD8+ T cells of similar phenotype in PBMCs.

  11. Boosting antitumor responses of T lymphocytes infiltrating human prostate cancers.

    PubMed

    Bronte, Vincenzo; Kasic, Tihana; Gri, Giorgia; Gallana, Keti; Borsellino, Giovanna; Marigo, Ilaria; Battistini, Luca; Iafrate, Massimo; Prayer-Galetti, Tommaso; Pagano, Francesco; Viola, Antonella

    2005-04-18

    Immunotherapy may provide valid alternative therapy for patients with hormone-refractory metastatic prostate cancer. However, if the tumor environment exerts a suppressive action on antigen-specific tumor-infiltrating lymphocytes (TIL), immunotherapy will achieve little, if any, success. In this study, we analyzed the modulation of TIL responses by the tumor environment using collagen gel matrix-supported organ cultures of human prostate carcinomas. Our results indicate that human prostatic adenocarcinomas are infiltrated by terminally differentiated cytotoxic T lymphocytes that are, however, in an unresponsive status. We demonstrate the presence of high levels of nitrotyrosines in prostatic TIL, suggesting a local production of peroxynitrites. By inhibiting the activity of arginase and nitric oxide synthase, key enzymes of L-arginine metabolism that are highly expressed in malignant but not in normal prostates, reduced tyrosine nitration and restoration of TIL responsiveness to tumor were achieved. The metabolic control exerted by the tumor on TIL function was confirmed in a transgenic mouse prostate model, which exhibits similarities with human prostate cancer. These results identify a novel and dominant mechanism by which cancers induce immunosuppression in situ and suggest novel strategies for tumor immunotherapy.

  12. Tumor-Infiltrating Lymphocytes in Triple Negative Breast Cancer: The Future of Immune Targeting

    PubMed Central

    García-Teijido, Paula; Cabal, María Luque; Fernández, Ignacio Peláez; Pérez, Yolanda Fernández

    2016-01-01

    Triple negative breast cancer (TNBC) is a highly heterogeneous tumor. There is increasing evidence of the role of tumor lymphocytic immune infiltrates in this subtype of breast cancer. Robust levels of tumor infiltrating lymphocytes (TILs) have been associated with improved disease-free and overall survival rates in TNBC patients with and without any treatment. Recent efforts have been made to develop a standardized methodology for evaluating TILs. The presence of TILs in the breast tumor microenvironment can also predict responses not only to neoadjuvant but also to adjuvant chemotherapy treatments. High numbers of TILs correlate with increased pathological complete responses (pCR) in TNBC. TILs are prognostic and predictive of response to standard therapies; thus, the immune system appears to play an active role in a subgroup of breast cancer. There is an increasing interest in directly targeting the immune system as part of breast cancer therapy, mainly in patients with TNBC. New immune modulatory agents, including immune checkpoints inhibitors, have shown promising activity in a subgroup of metastatic TNBC. Increased programmed cell death protein 1 ligand (PD-L1) expression on the surface of TNBC provides the rationale for implementing therapeutic strategies targeting the PD-1/PD-L1 axis in TNBC. The programmed cell death protein 1 (PD-1) inhibitor pembrolizumab, and the PD-L1 inhibitor atezolizumab have shown promising results in clinical trials. PMID:27081325

  13. Scaffold functions of 14-3-3 adaptors in B cell immunoglobulin class switch DNA recombination.

    PubMed

    Lam, Tonika; Thomas, Lisa M; White, Clayton A; Li, Guideng; Pone, Egest J; Xu, Zhenming; Casali, Paolo

    2013-01-01

    Class switch DNA recombination (CSR) of the immunoglobulin heavy chain (IgH) locus crucially diversifies antibody biological effector functions. CSR involves the induction of activation-induced cytidine deaminase (AID) expression and AID targeting to switch (S) regions by 14-3-3 adaptors. 14-3-3 adaptors specifically bind to 5'-AGCT-3' repeats, which make up for the core of all IgH locus S regions. They selectively target the upstream and downstream S regions that are set to undergo S-S DNA recombination. We hypothesized that 14-3-3 adaptors function as scaffolds to stabilize CSR enzymatic elements on S regions. Here we demonstrate that all seven 14-3-3β, 14-3-3ε, 14-3-3γ, 14-3-3η, 14-3-3σ, 14-3-3τ and 14-3-3ζ adaptors directly interacted with AID, PKA-Cα (catalytic subunit) and PKA-RIα (regulatory inhibitory subunit) and uracil DNA glycosylase (Ung). 14-3-3 adaptors, however, did not interact with AID C-terminal truncation mutant AIDΔ(180-198) or AIDF193A and AIDL196A point-mutants (which have been shown not to bind to S region DNA and fail to mediate CSR). 14-3-3 adaptors colocalized with AID and replication protein A (RPA) in B cells undergoing CSR. 14-3-3 and AID binding to S region DNA was disrupted by viral protein R (Vpr), an accessory protein of human immunodeficiency virus type-1 (HIV-1), which inhibited CSR without altering AID expression or germline IH-CH transcription. Accordingly, we demonstrated that 14-3-3 directly interact with Vpr, which in turn, also interact with AID, PKA-Cα and Ung. Altogether, our findings suggest that 14-3-3 adaptors play important scaffold functions and nucleate the assembly of multiple CSR factors on S regions. They also show that such assembly can be disrupted by a viral protein, thereby allowing us to hypothesize that small molecule compounds that specifically block 14-3-3 interactions with AID, PKA and/or Ung can be used to inhibit unwanted CSR.

  14. Life without dUTPase

    PubMed Central

    Kerepesi, Csaba; Szabó, Judit E.; Papp-Kádár, Veronika; Dobay, Orsolya; Szabó, Dóra; Grolmusz, Vince; Vértessy, Beáta G.

    2016-01-01

    Fine-tuned regulation of the cellular nucleotide pools is indispensable for faithful replication of Deoxyribonucleic Acid (DNA). The genetic information is also safeguarded by DNA damage recognition and repair processes. Uracil is one of the most frequently occurring erroneous bases in DNA; it can arise from cytosine deamination or thymine-replacing incorporation. Two enzyme activities are primarily involved in keeping DNA uracil-free: dUTPase (dUTP pyrophosphatase) activity that prevent thymine-replacing incorporation and uracil-DNA glycosylase activity that excise uracil from DNA and initiate uracil-excision repair. Both dUTPase and the most efficient uracil-DNA glycosylase (UNG) is thought to be ubiquitous in free-living organisms. In the present work, we have systematically investigated the genotype of deposited fully sequenced bacterial and Archaeal genomes. We have performed bioinformatic searches in these genomes using the already well described dUTPase and UNG gene sequences. For dUTPases, we have included the trimeric all-beta and the dimeric all-alpha families and also, the bifunctional dCTP (deoxycytidine triphosphate) deaminase-dUTPase sequences. Surprisingly, we have found that in contrast to the generally held opinion, a wide number of bacterial and Archaeal species lack all of the previously described dUTPase gene(s). The dut– genotype is present in diverse bacterial phyla indicating that loss of this (or these) gene(s) has occurred multiple times during evolution. We discuss potential survival strategies in lack of dUTPases, such as simultaneous lack or inhibition of UNG and possession of exogenous or alternate metabolic enzymes involved in uracil-DNA metabolism. The potential that genes previously not associated with dUTPase activity may still encode enzymes capable of hydrolyzing dUTP is also discussed. Our data indicate that several unicellular microorganisms may efficiently cope with a dut– genotype lacking all of the previously described d

  15. Human spontaneous labor without histologic chorioamnionitis is characterized by an acute inflammation gene expression signature

    PubMed Central

    Haddad, Ramsi; Tromp, Gerard; Kuivaniemi, Helena; Chaiworapongsa, Tinnakorn; Kim, Yeon Mee; Mazor, Moshe; Romero, Roberto

    2006-01-01

    OBJECTIVE The purpose of this study was to identify which biological processes may be involved in normal labor. STUDY DESIGN Transcriptional profiles for chorioamniotic membranes (n=24) and blood (n=20) were generated from patients at term with no labor (TNL) and in labor (TIL). RESULTS Expression of 197 transcripts (P≤0.02) differentiated TIL and TNL chorioamniotic membrane samples. Gene Ontology analysis indicated that TIL samples had increased expression of multiple chemokines and transcripts associated with neutrophil and monocyte recruitment. Microarray results were verified using quantitative real-time RT-PCR with independent samples. Transcriptional profiles from blood RNA revealed no Gene Ontology category enrichment of discriminant probe sets. CONCLUSION Labor induces gene expression changes consistent with localized inflammation, despite the absence of histologically detectable inflammation. PMID:16890549

  16. Production in vitro of thyroglobulin autoantibody by obese strain (OS) chickens.

    PubMed Central

    Tempelis, C H; Schauenstein, K; Wick, G

    1987-01-01

    Thyroglobulin autoantibody (Tg-AAb) can be spontaneously produced in vitro with thyroid infiltrating lymphocytes (TIL) collected from Obese strain chickens 3.5 and 4 weeks old. Attempts to enhance Tg-AAb synthesis with two known polyclonal stimulators of immunoglobulin synthesis in chickens, Staphylococcus aureus Cowan strain 1 and dextran sulphate, failed to increase Tg-AAb production in vitro. Spleen cells and peripheral blood lymphocytes obtained from the same chickens as the TIL and older chickens known to produce moderate to high levels of Tg-AAb in vivo did not produce autoantibody either spontaneously or in the presence of polyclonal Ig stimulators with one exception. With this single, exceptional chicken we obtained a small amount of Tg-AAb produced in vitro with spleen cells. This suggests that in the OS chicken TIL, and to a much lesser extent, the spleen, contribute to the total Tg-AAb produced in this model of autoimmune thyroiditis. PMID:2440629

  17. A novel human truncated IL12rβ1-Fc fusion protein ameliorates experimental autoimmune encephalomyelitis via specific binding of p40 to inhibit Th1 and Th17 cell differentiation

    PubMed Central

    Wang, Xin; Luo, Cheng; Yu, Dongmei; Wang, Yuheng; Chen, Yucong; Lei, Wen; Gao, Xiangdong; Yao, Wenbing

    2015-01-01

    Interleukin (IL)-12 and IL-23 respectively driving polarization of T helper (Th) 1 and Th17 cells has been strongly implicated in the pathogenesis of both multiple sclerosis (MS) and experimental autoimmune encephalomyelitis (EAE). In this study, we first constructed, expressed and purified a novel human truncated IL12rβ1-Fc fusion protein (tIL12rβ1/Fc) binding multiple forms of the p40 subunit of human IL-12 and IL-23. tIL12rβ1/Fc was found to effectively ameliorate MOG35–55-induced EAE through reducing the production of Th1- and Th17-polarized pro-inflammatory cytokines and suppressing inflammation and demyelination in the focused parts. Moreover, tIL12rβ1/Fc suppressed Th1 (IFN-γ+ alone) and IFN-γ+ IL-17+ as well as the population of classic Th17 (IL-17+ alone) cells in vivo. Furthermore, tIL12rβ1/Fc ameliorated EAE at the peak of disease via the inhibition of STAT pathway, thereby causing a prominent reduction of RORγt (Th17) and T-bet (Th1) expression. Notably, tIL12rβ1/Fc could increase the relative number of CD4+ Foxp3+ regulatory T cells. These findings indicates that tIL12rβ1/Fc is a novel fusion protein for specific binding multiple forms of p40 subunit to exert potent anti-inflammatory effects and provides a valuable approach for the treatment of MS and other autoimmune diseases. PMID:26384304

  18. How does breathing frequency affect the performance of an N95 filtering facepiece respirator and a surgical mask against surrogates of viral particles?

    PubMed

    He, Xinjian; Reponen, Tiina; McKay, Roy; Grinshpun, Sergey A

    2014-01-01

    Breathing frequency (breaths/min) differs among individuals and levels of physical activity. Particles enter respirators through two principle penetration pathways: faceseal leakage and filter penetration. However, it is unknown how breathing frequency affects the overall performance of N95 filtering facepiece respirators (FFRs) and surgical masks (SMs) against viral particles, as well as other health-relevant submicrometer particles. A FFR and SM were tested on a breathing manikin at four mean inspiratory flows (MIFs) (15, 30, 55, and 85 L/min) and five breathing frequencies (10, 15, 20, 25, and 30 breaths/min). Filter penetration (Pfilter) and total inward leakage (TIL) were determined for the tested respiratory protection devices against sodium chloride (NaCl) aerosol particles in the size range of 20 to 500 nm. "Faceseal leakage-to-filter" (FLTF) penetration ratios were calculated. Both MIF and breathing frequency showed significant effects (p < 0.05) on Pfilter and TIL. Increasing breathing frequency increased TIL for the N95 FFR whereas no clear trends were observed for the SM. Increasing MIF increased Pfilter and decreased TIL resulting in decreasing FLTF ratio. Most of FLTF ratios were >1, suggesting that the faceseal leakage was the primary particle penetration pathway at various breathing frequencies. Breathing frequency is another factor (besides MIF) that can significantly affect the performance of N95 FFRs, with higher breathing frequencies increasing TIL. No consistent trend of increase or decrease of TIL with either MIF or breathing frequency was observed for the tested SM. To potentially extend these findings beyond the manikin/breathing system used, future studies are needed to fully understand the mechanism causing the breathing frequency effect on the performance of respiratory protection devices on human subjects.

  19. Immunomodulation by MYB is associated with tumor relapse in patients with early stage colorectal cancer

    PubMed Central

    Millen, Rosemary; Malaterre, Jordane; Cross, Ryan S.; Carpinteri, Sandra; Desai, Jayesh; Tran, Ben; Darcy, Phillip; Gibbs, Peter; Sieber, Oliver; Zeps, Nikolajs; Waring, Paul; Fox, Stephen; Pereira, Lloyd; Ramsay, Robert G.

    2016-01-01

    ABSTRACT The presence of tumor immune infiltrating cells (TILs), particularly CD8+ T-cells, is a robust predictor of outcome in patients with colorectal cancer (CRC). We revisited TIL abundance specifically in patients with microsatellite stable (MSS) CRC without evidence of lymph node or metastatic spread. Examination of the density of CD8+ T-cells in primary tumors in the context of other pro-oncogenic markers was performed to investigate potential regulators of TILs. Two independent cohorts of patients with MSS T2-4N0M0 CRC, enriched for cases with atypical relapse, were investigated. We quantified CD8+ and CD45RO+ -TILs, inflammatory markers, NFkBp65, pStat3, Cyclo-oxygenase-2 (COX2) and GRP78 as well as transcription factors (TF), β-catenin and MYB. High CD8+ TILs correlated with a better relapse-free survival in both cohorts (p = 0.002) with MYB and its target gene, GRP78 being higher in the relapse group (p = 0.001); no difference in pSTAT3 and p65 was observed. A mouse CRC (CT26) model was employed to evaluate the effect of MYB on GRP78 expression as well as T-cell infiltration. MYB over-expressing in CT26 cells increased GRP78 expression and the analysis of tumor-draining lymph nodes adjacent to tumors showed reduced T-cell activation. Furthermore, MYB over-expression reduced the efficacy of anti-PD-1 to modulate CT26 tumor growth. This high MYB and GRP78 show a reciprocal relationship with CD8+ TILs which may be useful refining the prediction of patient outcome. These data reveal a new immunomodulatory function for MYB suggesting a basis for further development of anti-GRP78 and/or anti-MYB therapies. PMID:27622014

  20. Enthalpic and entropic contributions mediate the role of disulfide bonds on the conformational stability of Interleukin-4

    PubMed Central

    Vaz, Daniela C.; Rodrigues, J. Rui; Sebald, Walter; Dobson, Christopher M.; Brito, Rui M.M.

    2006-01-01

    The role of disulfide bridges in the structure, stability, and folding pathways of proteins has been the subject of wide interest in the fields of protein design and engineering. However, the relative importance of entropic and enthalpic contributions for the stabilization of proteins provided by disulfides is not always clear. Here, we perform a detailed analysis of the role of disulfides in the conformational stability of human Interleukin-4 (IL4), a four-helix bundle protein. In order to evaluate the contribution of two out of the three disulfides to the structure and stability of IL4, two IL4 mutants, C3T-IL4 and C24T-IL4, were used. NMR and ANS binding experiments were compatible with altered dynamics and an increase of the nonpolar solvent-accessible surface area of the folded state of the mutant proteins. Chemical and thermal unfolding experiments followed by fluorescence and circular dichroism revealed that both mutant proteins have lower conformational stability than the wild-type protein. Transition temperatures of unfolding decreased 14°C for C3T-IL4 and 10°C for C24T-IL4, when compared to WT-IL4, and the conformational stability, at 25°C, decreased 4.9 kcal/mol for C3T-IL4 and 3.2 kcal/mol for C24T-IL4. Interestingly, both the enthalpy and the entropy of unfolding, at the transition temperature, decreased in the mutant proteins. Moreover, a smaller change in heat capacity of unfolding was also observed for the mutants. Thus, disulfide bridges in IL4 play a critical role in maintaining the thermodynamic stability and core packing of the helix bundle. PMID:16373475

  1. The expressions of MIF and CXCR4 protein in tumor microenvironment are adverse prognostic factors in patients with esophageal squamous cell carcinoma

    PubMed Central

    2013-01-01

    Background Tumor-derived cytokines and their receptors usually take important roles in the disease progression and prognosis of cancer patients. In this survey, we aimed to detect the expression levels of MIF and CXCR4 in different cell populations of tumor microenvironments and their association with survivals of patients with esophageal squamous cell carcinoma (ESCC). Methods MIF and CXCR4 levels were measured by immunochemistry in tumor specimens from 136 resected ESCC. Correlation analyses and independent prognostic outcomes were determined using Pearson’s chi-square test and Cox regression analysis. Results The expression of CXCR4 in tumor cells was positively associated with tumor status (P = 0.045) and clinical stage (P = 0.044); whereas the expression of CXCR4 in tumor-infiltrating lymphocytes (TILs) and the expression of MIF in tumor cells and in TILs were not associated with clinical parameters of ESCC patients. High MIF expression in tumor cells or in TILs or high CXCR4 expression in tumor cells was significantly related to poor survival of ESCC patients (P < 0.05). Multivariate analysis showed that the expression of MIF or CXCR4 in tumor cells and the expression of MIF in TILs were adverse independent factors for disease-free survival (DFS) and overall survival (OS) in the whole cohort of patients (P < 0.05). Furthermore, the expression of MIF and CXCR4 in tumor cells were independent factors for reduced DFS and OS in metastatic/recurrent ESCC patients (P < 0.05). Interestingly, the expressions of MIF and CXCR4 in tumor cells and in TILs were significantly positively correlated (P < 0.05), and the combined MIF and CXCR4 expression in tumor cells was an independent adverse predictive factor for DFS and OS (P < 0.05). Conclusion The expressions of MIF and CXCR4 proteins in tumor cells and TILs have different clinically predictive values in ESCC. PMID:23497377

  2. PD-1 identifies the patient-specific CD8⁺ tumor-reactive repertoire infiltrating human tumors.

    PubMed

    Gros, Alena; Robbins, Paul F; Yao, Xin; Li, Yong F; Turcotte, Simon; Tran, Eric; Wunderlich, John R; Mixon, Arnold; Farid, Shawn; Dudley, Mark E; Hanada, Ken-Ichi; Almeida, Jorge R; Darko, Sam; Douek, Daniel C; Yang, James C; Rosenberg, Steven A

    2014-05-01

    Adoptive transfer of tumor-infiltrating lymphocytes (TILs) can mediate regression of metastatic melanoma; however, TILs are a heterogeneous population, and there are no effective markers to specifically identify and select the repertoire of tumor-reactive and mutation-specific CD8⁺ lymphocytes. The lack of biomarkers limits the ability to study these cells and develop strategies to enhance clinical efficacy and extend this therapy to other malignancies. Here, we evaluated unique phenotypic traits of CD8⁺ TILs and TCR β chain (TCRβ) clonotypic frequency in melanoma tumors to identify patient-specific repertoires of tumor-reactive CD8⁺ lymphocytes. In all 6 tumors studied, expression of the inhibitory receptors programmed cell death 1 (PD-1; also known as CD279), lymphocyte-activation gene 3 (LAG-3; also known as CD223), and T cell immunoglobulin and mucin domain 3 (TIM-3) on CD8⁺ TILs identified the autologous tumor-reactive repertoire, including mutated neoantigen-specific CD8⁺ lymphocytes, whereas only a fraction of the tumor-reactive population expressed the costimulatory receptor 4-1BB (also known as CD137). TCRβ deep sequencing revealed oligoclonal expansion of specific TCRβ clonotypes in CD8⁺PD-1⁺ compared with CD8⁺PD-1- TIL populations. Furthermore, the most highly expanded TCRβ clonotypes in the CD8⁺ and the CD8⁺PD-1⁺ populations recognized the autologous tumor and included clonotypes targeting mutated antigens. Thus, in addition to the well-documented negative regulatory role of PD-1 in T cells, our findings demonstrate that PD-1 expression on CD8⁺ TILs also accurately identifies the repertoire of clonally expanded tumor-reactive cells and reveal a dual importance of PD-1 expression in the tumor microenvironment.

  3. Campus Single Sign-On und hochschulübergreifendes Identity Management

    NASA Astrophysics Data System (ADS)

    Hommel, Wolfgang

    Das im Rahmen von IntegraTUM für die TUM geschaffene Identity & Access Management System setzt das Paradigma unified login um, d. h. ein Benutzer kann alle für ihn relevanten Dienste innerhalb der Hochschule mit derselben Loginname-/Passwortkombination nutzen. Dieser Artikel zeigt, wie auf Basis der Software Shibboleth und der deutschlandweiten Hochschulföderation DFN-AAI als weitere Mehrwerte das campusweite web single sign-on und die nahtlose Nutzung zahlreicher externer Web-Anwendungen erreicht werden. Als Beispiel für die Abläufe bei der Erschließung neuer Dienste für die hochschulübergreifende Nutzung wird die Anbindung von Learning Management Systemen auf Basis des DFN-AAI E-Learning-Profils diskutiert. Den umfassenden Vorteilen werden schließlich die aktuellen technischen Grenzen bei der Umsetzung des hochschulübergreifenden Identity Management gegenübergestellt.

  4. National Dam Safety Program. Alexander Lake Dam (Inventory Number N.Y. 936), Susquehanna River Basin, Tioga County, New York. Phase I Inspection Report,

    DTIC Science & Technology

    1981-07-01

    sasi xwmwv*Aa "a onomwvf -t 4,tLft "S蓸 0 doll?1 wi #m$rooa w WOW I -stm * r &4 gwff t(IU 14640" asa.u a4 . n.vunu .3 NOWc M f IANO V" 4’IV w- domin C...81 L D ANDERSEN DACWSI-81-C-O01D UNCLASSIFIED NL *NNnnnnnlInSIhhhhhIEE BlIIIIIIIIIIIl I IIIIhhhhhhh 111111-0 13 1w 111140 MICROCO~PY R [SOLUION IS...NEW YORK DISTRICT CORPS OF ENGINEERS__J JULY 1981 APPROVED FOR PUBLIC RELE.’ I DISTR!BUTION UNgM.rED 0 19 82052 St 7 e r (:&. A f 136olf T N1 S ft

  5. [Educational competence of parents with children participating in youth welfare measures].

    PubMed

    Rücker, Stefan; Büttner, Peter; Petermann, Ulrike; Petermann, Franz

    2013-07-01

    Fragestellung: In der vorliegenden Studie wird der Einfluss ausgewählter Risiken auf das Erziehungsverhalten von Eltern am Beginn einer Jugendhilfe-Maßnahme analysiert. Methodik: Familiäre Belastungen sowie Erziehungsdimensionen wurden anhand standardisierter Erhebungsverfahren bei N = 74 Eltern erfasst. Ergebnisse: In Abhängigkeit des Belastungsausmaßes konnten drei Gruppen gebildet werden. Die Befragungsergebnisse wurden im Querschnitt varianzanalytisch ausgewertet. Familien mit spezifischen Risiken, wie materielle Belastungen, psychisch erkrankte Elternteile oder einen Alleinerziehendenstatus weisen signifikant ungünstigere Werte bei Erziehungsmerkmalen auf. In negativen Erziehungsmerkmalen kann der Einfluss spezifischer Risiken statistisch nicht abgesichert werden. Schlussfolgerungen: Spezifisch belastete Eltern in Jugendhilfe-Maßnahmen benötigen eventuell ein spezielles Training zur Steigerung ihrer Erziehungskompetenz. Dies gilt vor allem dann, wenn Prä-Post-Vergleiche am Ende der Maßnahme ein geringeres Erziehungs-Outcome in der spezifisch belasteten Gruppe offenbaren.

  6. [Acanthamoeba keratitis after use of soft contact lenses--case report].

    PubMed

    Ziak, P; Ondriska, F; Mrva, M

    2003-09-01

    The case history of a 39-year patient suffering from a deep inflammation of cornea and not responding to conventional antibiotic treatment is presented. The patient was using soft contact lenses during the period of initial symptoms; moreover, he was bathing in thermal bathing pool. A cultivation examination of smears from the area of corneal defect revealed the presence of Acanthamoeba lugdunensis in combination with bacterial infection by Pseudomonas aeruginosa. The available data indicate that it is the first case of acanthamoeba karatitis (AK) after the application of contact lenses in Slovakia. A long-term local treatment with propamidin isethionate (Brolene gtt, ung.) resulted in healing up. The subsequent vision after 16 months since the initial symptoms proved to be 6/12 (0.5). The healing of the centrally localized defect changed the curvature of cornea with consequent hypermetropic shift. The defect completely corrected the patient's myopia (-8.5). The paper describes present possibilities of AK therapy.

  7. Guidelines and Data to Support Plans for Reallocating Food during Crisis Relocation. Regional Appendix. FEMA Region IV,

    DTIC Science & Technology

    1982-09-01

    distri- butors serving the region, arid a list of counties served by each distributor is included for reference. U TOP TEN FOOD DISTRIBUTORSco D SERVING...SUPER VALU STORES INC XENIA Oil 122,750 28 4,:84 HOPKINS MN 10,000 1 10,000 SUPER VALU STORES HOPKINS MN 5,000 1 5,000 r p p C-3 LIST OF COUNTIES SERVED...BY EACH DISTRIBUTOR IN FEMA REGION IV 𔃻. 9 COUNTY SUPPLIERS TOTAL 500K- 1-2 2-4 4-8 6+ EST ACV STORES I MIL tIL MIL MIL tIL ($1000) REGION 041! STATE

  8. PD-1, PD-L1 Protein Expression in Non-Small Cell Lung Cancer and Their Relationship with Tumor-Infiltrating Lymphocytes

    PubMed Central

    He, Yayi; Rozeboom, Leslie; Rivard, Christopher J.; Ellison, Kim; Dziadziuszko, Rafał; Yu, Hui; Zhou, Caicun; Hirsch, Fred R.

    2017-01-01

    Background Immunotherapy targeting the programmed death-1 (PD-1)/programmed death ligand-1 (PD-L1) checkpoint has shown the good outcomes in non-small cell lung cancer (NSCLC). We investigated PD-1 and PD-L1 protein expression and their correlation with tumor-infiltrating lymphocytes (TILs), and association with survival in NSCLC. Material/Methods The expression of PD-1 (NAT105, Cell Marque) and PD-L1 (28-8, Dako) protein was assessed in 55 NSCLC cell lines by immunohistochemistry (IHC). PD-1 (NAT105, Cell Marque) and PD-L1 (22C3, Dako) protein expression was evaluated by IHC, and TIL percentage was scored, in 139 surgically resected specimens from patients with NSCLC. Results PD-1 was not expressed on NSCLC cell lines. PD-L1 was expressed on 20 NSCLC cell lines (36.4%). A total of 60 patient samples (43.2%) were positive for PD-1 on the TILs, and 25 (18.0%) were positive for PD-L1 on tumor cells. High expression of PD-1 on tumor cells was significantly correlated with higher expression of PD-L1 (P=0.026) and a higher percentage of TILs (P<0.001). In the Cox regression model, the odds ratio for PD-1 was 2.828 (95% CI: 1.325–11.165; P=0.013) and 8.579 (95% CI: 4.148–22.676; P<0.001) when PD-L1 and TILs were positive. Patients whose tumor cells were PD-L1 negative had a tendency for longer relapse-free survival (RFS) than patients who were PD-L1 positive (1.85 years, 95% CI: 0.77–2.93 vs. 0.97 years, 95% CI: 0.71–1.23; P=0.054). Conclusions PD-1 was expressed on TILs in tumor tissues in NSCLC patients. PD-L1 was expressed on both TILs and tumor tissues. PD-1 expression was correlated with PD-L1 on tumor cells and TILs. Patients who were PD-L1 positive tended to experience progression after surgery. PMID:28275222

  9. Phenotypic and functional analysis of tumor-infiltrating lymphocytes compared with tumor-associated lymphocytes from ascitic fluid and peripheral blood lymphocytes in patients with advanced ovarian cancer.

    PubMed

    Santin, A D; Hermonat, P L; Ravaggi, A; Bellone, S; Roman, J J; Smith, C V; Pecorelli, S; Radominska-Pandya, A; Cannon, M J; Parham, G P

    2001-01-01

    To investigate and compare the phenotype and function of lymphocytes collected from patients harboring advanced ovarian cancer, leukocytes from peripheral blood (n = 18), ascitic fluid (n = 13) and tumor tissues (n = 13) were evaluated for the relative proportions of lymphocyte subsets, including CD3+, CD4+, CD8+, CD19+, CD56 and the early (CD25) and late (HLA-DR) activation markers on CD3+ T cells. The ability to synthesize type 1 cytokines (IFN-gamma and IL-2) and a type 2 cytokine (IL-4) was assessed by flow cytometry. In all patients, T cells (CD3+) were the major leukocyte population detected in each tissue, with CD4+ T cells being dominant in peripheral blood lymphocytes (PBL) and tumor-associated lymphocytes (TAL) but not in tumor-infiltrating lymphocytes (TIL) (CD4:CD8 ratios: 3.0 vs. 2.0 vs. 1.0, respectively). CD19+ lymphocytes (B cells) and CD56+ lymphocytes (NK cells) were significantly higher in PBL compared to TAL and TIL (p < 0.05). TAL and TIL had a higher proportion of T cells expressing the late activation marker HLA-DR compared to PBL. In contrast, no significant differences were detected in PBL, TAL and TIL in the expression of the early activation marker CD25. Type 1 cytokines were the dominant type produced by in vitro stimulated T cells for each population, with a greater proportion of IFN-gamma+ T cells in TAL and TIL compared to PBL (p < 0.01), and a higher proportion of IL-2+ T cells in PBL compared with TAL and TIL (p < 0.05). Low percentages of IL-4+ T cells (i.e. Th2) were detected in each tissue. Taken together, these data demonstrate the recruitment and accumulation of high concentrations of antigen-experienced T lymphocytes in TAL and TIL compared to PBL. However, low surface expression of IL-2 receptor (i.e. CD25), as well as depressed intracellular IL-2 production in chronically stimulated TAL and TIL suggests that the impaired antitumor function commonly detected in these lymphocyte populations may be secondary to an acquired

  10. Differential expression of APE1 and APE2 in germinal centers promotes error-prone repair and A:T mutations during somatic hypermutation.

    PubMed

    Stavnezer, Janet; Linehan, Erin K; Thompson, Mikayla R; Habboub, Ghaith; Ucher, Anna J; Kadungure, Tatenda; Tsuchimoto, Daisuke; Nakabeppu, Yusaku; Schrader, Carol E

    2014-06-24

    Somatic hypermutation (SHM) of antibody variable region genes is initiated in germinal center B cells during an immune response by activation-induced cytidine deaminase (AID), which converts cytosines to uracils. During accurate repair in nonmutating cells, uracil is excised by uracil DNA glycosylase (UNG), leaving abasic sites that are incised by AP endonuclease (APE) to create single-strand breaks, and the correct nucleotide is reinserted by DNA polymerase β. During SHM, for unknown reasons, repair is error prone. There are two APE homologs in mammals and, surprisingly, APE1, in contrast to its high expression in both resting and in vitro-activated splenic B cells, is expressed at very low levels in mouse germinal center B cells where SHM occurs, and APE1 haploinsufficiency has very little effect on SHM. In contrast, the less efficient homolog, APE2, is highly expressed and contributes not only to the frequency of mutations, but also to the generation of mutations at A:T base pair (bp), insertions, and deletions. In the absence of both UNG and APE2, mutations at A:T bp are dramatically reduced. Single-strand breaks generated by APE2 could provide entry points for exonuclease recruited by the mismatch repair proteins Msh2-Msh6, and the known association of APE2 with proliferating cell nuclear antigen could recruit translesion polymerases to create mutations at AID-induced lesions and also at A:T bp. Our data provide new insight into error-prone repair of AID-induced lesions, which we propose is facilitated by down-regulation of APE1 and up-regulation of APE2 expression in germinal center B cells.

  11. Molecular Characterization of Type-Specific Capsular Polysaccharide Biosynthesis Genes of Streptococcus agalactiae Type Ia

    PubMed Central

    Yamamoto, Shin; Miyake, Katsuhide; Koike, Yoichi; Watanabe, Masaki; Machida, Yuichi; Ohta, Michio; Iijima, Shinji

    1999-01-01

    The type-specific capsular polysaccharide (CP) of a group B streptococcus, Streptococcus agalactiae type Ia, is a high-molecular-weight polymer consisting of the pentasaccharide repeating unit 4)-[α-d-NeupNAc-(2→3)-β-d-Galp-(1→4)-β-d-GlcpNAc-(1→3)]-β-d-Galp-(1→4)-β-d-Glcp-(1. Here, cloning, sequencing, and transcription of the type Ia-specific capsular polysaccharide synthesis (cps) genes and functional analysis of these gene products are described. A 26-kb DNA fragment containing 18 complete open reading frames (ORFs) was cloned. These ORFs were designated cpsIaA to cpsIaL, neu (neuraminic acid synthesis gene) A to D, orf1 and ung (uracil DNA glycosylase). The cps gene products of S. agalactiae type Ia were homologous to proteins involved in CP synthesis of S. agalactiae type III and S. pneumoniae serotype 14. Unlike the cps gene cluster of S. pneumoniae serotype 14, transcription of this operon may start from cpsIaA, cpsIaE, and orf1 because putative promoter sequences were found in front of these genes. Northern hybridization, reverse transcription-PCR, and primer extension analyses supported this hypothesis. DNA sequence analysis showed that there were two transcriptional terminators in the 3′ end of this operon (downstream of orf1 and ung). The functions of CpsIaE, CpsIaG, CpsIaI, and CpsIaJ were examined by glycosyltransferase assay by using the gene products expressed in Escherichia coli JM109 harboring plasmids containing various S. agalactiae type Ia cps gene fragments. Enzyme assays suggested that the gene products of cpsIaE, cpsIaG, cpsIaI, and cpsIaJ are putative glucosyltransferase, β-1,4-galactosyltransferase, β-1,3-N-acetylglucosaminyltransferase, and β-1,4-galactosyltransferase, respectively. PMID:10464185

  12. A Regulatory Role for NBS1 in Strand-Specific Mutagenesis during Somatic Hypermutation

    PubMed Central

    Du, Likun; Dunn-Walters, Deborah K.; Chrzanowska, Krystyna H.; Stankovic, Tanja; Kotnis, Ashwin; Li, Xin; Lu, Jiayi; Eggertsen, Gösta; Brittain, Claire; Popov, Sergey W.; Gennery, Andrew R.; Taylor, A. Malcolm R.; Pan-Hammarström, Qiang

    2008-01-01

    Activation-induced cytidine deaminase (AID) is believed to initiate somatic hypermutation (SHM) by deamination of deoxycytidines to deoxyuridines within the immunoglobulin variable regions genes. The deaminated bases can subsequently be replicated over, processed by base excision repair or mismatch repair, leading to introduction of different types of point mutations (G/C transitions, G/C transversions and A/T mutations). It is evident that the base excision repair pathway is largely dependent on uracil-DNA glycosylase (UNG) through its uracil excision activity. It is not known, however, which endonuclease acts in the step immediately downstream of UNG, i.e. that cleaves at the abasic sites generated by the latter. Two candidates have been proposed, an apurinic/apyrimidinic endonuclease (APE) and the Mre11-Rad50-NBS1 complex. The latter is intriguing as this might explain how the mutagenic pathway is primed during SHM. We have investigated the latter possibility by studying the in vivo SHM pattern in B cells from ataxia-telangiectasia-like disorder (Mre11 deficient) and Nijmegen breakage syndrome (NBS1 deficient) patients. Our results show that, although the pattern of mutations in the variable heavy chain (VH) genes was altered in NBS1 deficient patients, with a significantly increased number of G (but not C) transversions occurring in the SHM and/or AID targeting hotspots, the general pattern of mutations in the VH genes in Mre11 deficient patients was only slightly altered, with an increased frequency of A to C transversions. The Mre11-Rad50-NBS1 complex is thus unlikely to be the major nuclease involved in cleavage of the abasic sites during SHM, whereas NBS1 might have a specific role in regulating the strand-biased repair during phase Ib mutagenesis. PMID:18575580

  13. 78 FR 35290 - Government-Owned Inventions; Availability for Licensing

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-06-12

    ... attacking TIL from a patient's tumor based on these markers prior to re-infusion, in vitro culture time is... specific cell surface markers will reduce the culture time for these T cells before re-infusion into the... for autologous tumor recognition before re- infusion into the patient Development Stage:...

  14. Estimating soil organic carbon using aerial imagery and soil surveys

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Widespread implementation of precision agriculture practices requires low-cost, high-quality, georeferenced soil organic carbon (SOC) maps, but currently these maps require expensive sample collection and analysis. Widely available aerial imagery is a low-cost source of georeferenced data. After til...

  15. Hydrogeological analysis of the upper Dupi Tila Aquifer, towards the implementation of a managed aquifer-recharge project in Dhaka City, Bangladesh

    NASA Astrophysics Data System (ADS)

    Rahman, Mohammad Azizur; Wiegand, Bettina A.; Badruzzaman, A. B. M.; Ptak, Thomas

    2013-08-01

    A preliminary feasibility assessment of managed aquifer-recharge (MAR) techniques was undertaken for Dhaka City, Bangladesh. Considering the top impermeable-layer (TIL) thickness and the land-use classification, four primary MAR techniques have been suggested: (1) soil-aquifer treatment (SAT) for TIL thickness 0-8 m, (2) cascade-type recharge trenches/pits for TIL thickness 9-30 m, (3) aquifer storage, transfer and recovery (ASR/ASTR) for TIL thickness 31-52 m, and (4) use of natural wetlands to recharge water collected from open spaces. The study suggests that recharge trenches and pits will be the most appropriate MAR techniques, which can be implemented in most parts of the recharge area (ca. 277 km2). In case of a recharge trench, the lower parts (15-20 m) that are in direct contact with the aquifer can be backfilled with biosand filters with a reactive layer containing metallic iron (Fe0) to offer pre-treatment of the infiltrated water. In addition to the suggested four techniques, the regional groundwater flow direction, from the northwest and northeast towards Dhaka City, may allow use of the aquifer as a natural treatment and transport medium for groundwater, if spreading basins are installed in the greater Dhaka area.

  16. US EPA, Pesticide Product Label, , 01/14/1994

    EPA Pesticide Factsheets

    2011-04-21

    ... t. (.-}iP ' . 1 , " . " . _1. :.!J::·'lt }'.'f· (r)) Cd[)ic's uf your final l:rinl,·,l }'Ji'clin" : efl 1" '.'-0 t· 1.~~JS;· t}lf:~ pr'Jducl for shipmet"!t. :·'f~l.~r to til('~ l\\-· ...

  17. Throw the Book at 'Em

    NASA Astrophysics Data System (ADS)

    Statler, Thomas S.

    1997-09-01

    Buchangst pervades college astronomy. Instructors fret that no textbook is quite right, authors and publishers get shot down when they try to innovate, students count the days 'til they can sell back their unfriendly tomes. But none of this is going to change unless we make it happen.

  18. Channel Protein Engineering: A Novel Approach towards the Molecular Dissection Determinants in Ligand-Regulated Channels

    DTIC Science & Technology

    1990-02-15

    Biochemical and Biophysical Research Communications 162...Soc. Neurosci. 15 (1):970a. (1989) Vol. 162, No. 1, 1989 BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS July 14, 1989 Pages 352-356 TIlE M25...Press. hic. All rights of reproduction in any form reserved. 352 Vol. 162, No. 1, 1989 BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS pli

  19. ASCD in Retrospect. Contributions to the History of the Association for Supervision and Curriculum Development.

    ERIC Educational Resources Information Center

    Van Til, William, Ed.

    Nine past presidents, the current president, and the executive director of the Association for Supervision and Curriculum Development (ASCD) contributed a chapter each to this history of ASCD and the fields it has represented since its founding in 1943. The book's editor, William Van Til, provides an introductory overview of the organization's…

  20. Remote sensing of soil tilage intensity in central Iowa

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Crop residues on the soil surface decrease soil erosion, increase soil organic matter, improve soil quality, and reduce the amount of nutrients and pesticides that reaches stream and rivers. Crop residue cover is often used to classify soil tillage intensity and assess the extent of conservation til...

  1. Closing the Student Achievement Gap in California's Elementary Schools: A Lead Teachers' Perspective on Transformational Instructional Leadership

    ERIC Educational Resources Information Center

    Hays, Kelli

    2010-01-01

    This study has tackled the thorny problem of closing the Student Achievement Gap (SAG) in California's elementary schools. To address that problem, an "Integrated" form of educational leadership called Transformational Instructional Leadership (TIL), a form grounded in "best practices" of Transformational and Instructional…

  2. Manipulations of soil microbiota for C sequestration and mitigation of greenhouse gas emissions in managed systems

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Soil microbes dominate processes that regulate soil trace gas emissions and soil C and N dynamics. Intensive management in agroecosystems provides unique opportunities to assess the effectiveness of microbial manipulations to enhance soil C retention and reduce trace gas emissions. While reduced til...

  3. US EPA, Pesticide Product Label, , 01/29/1996

    EPA Pesticide Factsheets

    2011-04-21

    ... tbt 11 Ut 1 :-, !lilt d i(aL~(;n " [lot con.'--) L<-;~l'Il·._ \\,1 til tit.' t f'l11l-i I)f f~ ~~ It let' (,- , md .j) Ill; ; ~:, J 1. tillS \\',n.itH t_ rll\\" III' j)-~ In vjrddtl"r1 . ...

  4. US EPA, Pesticide Product Label, FERTI-LOME TOMATO ...

    EPA Pesticide Factsheets

    2011-04-14

    ... {i{'Hii,=-~~ til :Y4 ~! t :fff":> (i1"ffi ~ rro:JT~ • ~ ill 111' foi ' I .' ' .. J' I'L"I"~ln,!J' \\ .f ."l.:JIl":U ~.H: ~wr) ::J;I:i~1 ;1if:i,;\\l)iJlmJ1!}.I~:\\ij1(ih~' 1 , + J II , . ...

  5. US EPA, Pesticide Product Label, METHYL BROMIDE 99.75% ...

    EPA Pesticide Factsheets

    2011-04-21

    ... ",alt.. Ind .. "t'l l.balaM., with ,robabla r.COyary .ft.r • parlod of na •• ,.,vr.. Ilood lIrO-l4. 1 • .".ls ... l~ ·Do "ot r ..... Ie .... _til ttl. -art.. T ••• t .... , MI'It".ra ... a1 ...

  6. US EPA, Pesticide Product Label, , 12/10/1986

    EPA Pesticide Factsheets

    2011-04-14

    ... (, I.; " " , e!',' ( fl II .~ \\~ . t .. te. ',!; I;!... t" .' t 1 i ., 'f. • :,1 t ; I'. I (., • I': l \\. , t ~ r 1 j. ,,-' tit '- I . l ti, l' j'(I..) .. I{l,!~!' c' ti:l !,. r{.;., t .. 1 • ...

  7. RAS/MAPK activation is associated with reduced tumor-infiltrating lymphocytes in triple-negative breast cancer: therapeutic cooperation between MEK and PD-1/PD-L1 immune checkpoint inhibitors

    PubMed Central

    Loi, Sherene; Dushyanthen, Sathana; Beavis, Paul A; Salgado, Roberto; Denkert, Carsten; Savas, Peter; Combs, Susan; Rimm, David L.; Giltnane, Jennifer M.; Estrada, Monica V.; Sánchez, Violeta; Sanders, Melinda E.; Cook, Rebecca S.; Pilkinton, Mark A.; Mallal, Simon A.; Wang, Kai; Miller, Vincent A.; Stephens, Phil J.; Yelensky, Roman; Doimi, Franco D.; Gómez, Henry; Ryzhov, Sergey V.; Darcy, Phillip K.; Arteaga, Carlos L.; Balko, Justin M.

    2015-01-01

    Purpose Tumor-infiltrating lymphocytes (TILs) in the residual disease (RD) of triple-negative breast cancers (TNBCs) after neoadjuvant chemotherapy (NAC) are associated with improved survival, but insight into tumor cell-autonomous molecular pathways affecting these features are lacking. Experimental Design We analyzed TILs in the RD of clinically and molecularly characterized TNBCs after NAC and explored therapeutic strategies targeting combinations of MEK inhibitors with PD-1/PD-L1-targeted immunotherapy in mouse models of breast cancer. Results Presence of TILs in the RD was significantly associated with improved prognosis. Genetic or transcriptomic alterations in Ras/MAPK signaling were significantly correlated with lower TILs. MEK inhibition up-regulated cell-surface major histocompatibility complex (MHC) expression and PD-L1 in TNBC cells both in vivo and in vitro. Moreover, combined MEK and PDL-1/PD-1 inhibition enhanced anti-tumor immune responses in mouse models of breast cancer. Conclusions These data suggest the possibility that Ras/MAPK pathway activation promotes immune-evasion in TNBC, and support clinical trials combining MEK- and PD-L1-targeted therapies. Furthermore, Ras/MAPK activation and MHC expression may be predictive biomarkers of response to immune checkpoint inhibitors. PMID:26515496

  8. Interlesional diversity of T cell receptors in melanoma with immune checkpoints enriched in tissue-resident memory T cells

    PubMed Central

    Boddupalli, Chandra Sekhar; Bar, Noffar; Kadaveru, Krishna; Krauthammer, Michael; Pornputtapong, Natopol; Ariyan, Stephan; Narayan, Deepak; Kluger, Harriet; Deng, Yanhong; Verma, Rakesh; Das, Rituparna; Bacchiocchi, Antonella; Halaban, Ruth; Sznol, Mario; Dhodapkar, Madhav V.; Dhodapkar, Kavita M.

    2016-01-01

    Heterogeneity of tumor cells and their microenvironment can affect outcome in cancer. Blockade of immune checkpoints (ICPs) expressed only on a subset of immune cells leads to durable responses in advanced melanoma. Tissue-resident memory T (TRM) cells have recently emerged as a distinct subset of memory T cells in nonlymphoid tissues. Here, we show that functional properties and expression of ICPs within tumor-infiltrating lymphocytes (TILs) differ from those of blood T cells. TILs secrete less IL-2, IFN-γ, and TNF-α compared with circulating counterparts, and expression of VEGF correlated with reduced TIL infiltration. Within tumors, ICPs are particularly enriched within T cells with phenotype and genomic features of TRM cells and the CD16+ subset of myeloid cells. Concurrent T cell receptor (TCR) and tumor exome sequencing of individual metastases in the same patient revealed that interlesional diversity of TCRs exceeded differences in mutation/neoantigen load in tumor cells. These findings suggest that the TRM subset of TILs may be the major target of ICP blockade and illustrate interlesional diversity of tissue-resident TCRs within individual metastases, which did not equilibrate between metastases and may differentially affect the outcome of immune therapy at each site. PMID:28018970

  9. US EPA, Pesticide Product Label, BIOBAN-C, 01/18/1971

    EPA Pesticide Factsheets

    2011-04-13

    ... HOB,\\1\\-S S'J(jiUfll PI'upi()natl' alld BI()BAl\\-C CalciLi:1 Pt'opiullate a 1'1' I •. , ( 'l' II l' II t asp I' I • S I' r vat ive ,.; tIl I' (' tar d ... Anti':lycl)tic al.!;l'llts als" "lJlJ ...

  10. The Desire to Learn as a Kind of Love: Gardening, Cooking, and Passion in Outdoor Education

    ERIC Educational Resources Information Center

    Wistoft, Karen

    2013-01-01

    "Gardens for Bellies" ["Haver til Maver"] is an organic school gardens project at Krogerup farm in Northern Sealand, Denmark, which provides children with first-hand experiences in a natural, outdoor environment. The general intention of the project is to expand children's competences and their knowledge of nature, farming and…

  11. Algunos Resumenes

    NASA Astrophysics Data System (ADS)

    1980-12-01

    EI "Palomar Observatory Sky Survey" es un medio auxiliar bien conocido y ütil para los astronomos. Todo el cielo dei hemisferio Norte esta captado en lotografias, cuyas reproducciones se encuentran archivadas en las bibliotecas de casi todos los observatorios importantes dei mundo.

  12. US EPA, Pesticide Product Label, METHYL BROMIDE 99.5% ...

    EPA Pesticide Factsheets

    2011-04-21

    ... to I.Rl In.t.ruc:tlo ..... co"ta(;t YOUif st.te ' •• Uetn <;til' r;n ... lra,..".l Control ""' • . or th. ... ... I ••••• control. cov.r ,., .. dl.tl, .flth ••••• 11.ht 1.,.,.ul1n b, ...

  13. Phenotypic gain from introgression of two QTLs, qSB9-2 and qSB12-1, for rice sheath blight resistance

    Technology Transfer Automated Retrieval System (TEKTRAN)

    F2:3 families from crosses between three indica introgression lines and their common japonica recurrent parent were used to evaluate two quantitative trait loci (QTLs) for sheath blight (SB) resistance. Three selected ‘TeQing’-into-‘Lemont’ backcross introgression lines (TILs) were more resistant th...

  14. Results of the Second U.S. Manned Suborbital Space Flight, July 21, 1961

    DTIC Science & Technology

    1961-07-21

    SPACECRAFT AND FLIGHT PLAN FOR THE MERCURY -REDSTONE 4 FLIGHT .............................................................. 3 By Jercme B. Haammack... Mercury -Redstone Project Engineer, NASA Manned Spacecraft Center. 3. RESULTS OF TIlE MR-4 PREFLIGHT AND POSTFLIGHT MEDICAL EXAMI- NATION CONDUCTED ON... MERCURY -REDSTONF MISSIONS 3 AND 4 ................................................................... 23 By William K. Douglas, M.D., Astronaut Fi

  15. Programming for Adolescents with Behavioral Disorders.

    ERIC Educational Resources Information Center

    Braaten, Sheldon, Ed.; And Others

    This book presents 17 papers from a 1982 national multidisciplinary conference on services for behaviorally disordered adolescents. The following papers are included: "Programming for Youth in Secondary Schools and the Community," (W. Van Til); "Who's Crazy? II" (C. Michael Nelson); "Correlates of Successful Adaptive Behavior: Comparative Studies…

  16. Predictive and Prognostic Role of Tumor-Infiltrating Lymphocytes for Early Breast Cancer According to Disease Subtypes: Sensitivity Analysis of Randomized Trials in Adjuvant and Neoadjuvant Setting

    PubMed Central

    Carbognin, Luisa; Pilotto, Sara; Nortilli, Rolando; Brunelli, Matteo; Nottegar, Alessia; Sperduti, Isabella; Giannarelli, Diana; Tortora, Giampaolo

    2016-01-01

    Background. The role of tumor-infiltrating lymphocytes (TILs) in breast cancer (BC) is still an issue for clinical research. Toward this end, a sensitivity analysis of neoadjuvant and adjuvant randomized clinical trials was performed according to disease subtypes. Methods. Pathological complete responses (pCRs) after neoadjuvant treatment according to the presence or absence of lymphocyte-predominant BC (LPBC) were extracted and cumulated as odds ratios (ORs) by adopting a random-effects model by subtype. Overall survival hazard ratios as a function of 10% incremental values of stromal TILs (sTILs) in adjuvant trials were extracted. The interaction test was adopted to determine the differential effect according to the subtype. Results. Eight trials (5,514 patients) were identified. With regard to neoadjuvant setting (4 studies), a significant interaction (p < .0001) according to LPBC was found. The presence of LPBC was associated with a 29.5% increase in pCR rate compared with non-LPBC (p < .0001). The pCR rate was significantly higher in patients with LPBC in triple-negative BC (TNBC) and HER2-positive BC settings, with an absolute difference of 15.7% (95% confidence interval [CI], 4.9%–26.2%) and 33.3% (95% CI, 23.6%–42.7%), respectively. With respect to the adjuvant setting (4 studies), a significant interaction (p < .0001) according to sTILs was found. A survival benefit was more likely to be determined for HER2-positive BC (p = .025) and TNBC (p < .0001), with no statistically significant difference for estrogen receptor-positive/HER2-negative disease. Conclusion. Despite the retrospective nature of this analysis, the presence of TILs may represent a robust predictive and prognostic marker for BC, particularly for TNBC and HER2-positive disease. Implications for Practice: This sensitivity analysis of neoadjuvant and adjuvant randomized clinical trials in breast cancer explores the potential predictive and prognostic role of tumor-infiltrating lymphocytes

  17. Effects of TP53 and PIK3CA mutations in early breast cancer: a matter of co-mutation and tumor-infiltrating lymphocytes.

    PubMed

    Kotoula, Vassiliki; Karavasilis, Vasilios; Zagouri, Flora; Kouvatseas, George; Giannoulatou, Eleni; Gogas, Helen; Lakis, Sotiris; Pentheroudakis, George; Bobos, Mattheos; Papadopoulou, Kyriaki; Tsolaki, Eleftheria; Pectasides, Dimitrios; Lazaridis, Georgios; Koutras, Angelos; Aravantinos, Gerasimos; Christodoulou, Christos; Papakostas, Pavlos; Markopoulos, Christos; Zografos, George; Papandreou, Christos; Fountzilas, George

    2016-07-01

    The purpose of this study is to investigate whether the outcome of breast cancer (BC) patients treated with adjuvant chemotherapy is affected by co-mutated TP53 and PIK3CA according to stromal tumor-infiltrating lymphocytes (TILs). Paraffin tumors of all clinical subtypes from 1661 patients with operable breast cancer who were treated within 4 adjuvant trials with anthracycline-taxanes chemotherapy were informative for TP53 and PIK3CA mutation status (semiconductor sequencing genotyping) and for stromal TILs density. Disease-free survival (DFS) was examined. TP53 mutations were associated with higher (p < 0.001) and PIK3CA with lower (p = 0.004) TILs in an ER /PgR-specific manner (p < 0.001). Mutations did not affect the favorable DFS of patients with lymphocyte-predominant (LP) BC. Within non-LPBC, PIK3CA-only mutations conferred best, while TP53-PIK3CA co-mutations (6 % of all tumors) conferred worst DFS (HR 0.59; 95 % CI 0.44-0.79; p = 0.001 for PIK3CA-only). TP53-only mutations were unfavorable in patients with lower TILs, while patients with lower TILs performed worse if their tumors carried TP53-only mutations (interaction p = 0.046). Multivariate analysis revealed favorable PIK3CA-only mutations in non-LPBC (HR 0.64; 95 % CI 0.47-0.88; p = 0.007), and unfavorable TP53 mutations in ER/PgRpos/HER2neg (HR 1.55; 95 % CI 1.07-2.24; p = 0.021). Mutations did not interact with TILs in non-LP triple-negative and HER2-positive patients. TP53 and PIK3CA mutations appear to have diverse effects on the outcome of early BC patients, according to whether these genes are co-mutated or not, and for TP53 according to TILs density and ER/PgR-status. These findings need to be considered when evaluating the effect of these two most frequently mutated genes in the context of large clinical trials.

  18. Association between Chemotherapy-Response Assays and Subsets of Tumor-Infiltrating Lymphocytes in Gastric Cancer: A Pilot Study

    PubMed Central

    Lee, Jee Youn; Son, Taeil; Cheong, Jae-Ho; Hyung, Woo Jin; Noh, Sung Hoon; Kim, Choong-Bai; Park, Chung-Gyu

    2015-01-01

    Purpose The purpose of this pilot study was to evaluate the association between adenosine triphosphate-based chemotherapy response assays (ATP-CRAs) and subsets of tumor infiltrating lymphocytes (TILs) in gastric cancer. Materials and Methods In total, 15 gastric cancer tissue samples were obtained from gastrectomies performed between February 2007 and January 2011. Chemotherapy response assays were performed on tumor cells from these samples using 11 chemotherapeutic agents, including etoposide, doxorubicin, epirubicin, mitomycin, 5-fluorouracil (5-FU), oxaliplatin, irinotecan, docetaxel, paclitaxel, methotrexate, and cisplatin. TILs in the tissue samples were evaluated using antibodies specific for CD3, CD4, CD8, Foxp3, and Granzyme B. Results The highest cancer cell death rates were induced by etoposide (44.8%), 5-FU (43.1%), and mitomycin (39.9%). Samples from 10 patients who were treated with 5-FU were divided into 5-FU-sensitive and -insensitive groups according to median cell death rate. No difference was observed in survival between the two groups (P=0.216). Only two patients were treated with a chemotherapeutic agent determined by an ATP-CRA and there was no significant difference in overall survival compared with that of patients treated with their physician's choice of chemotherapeutic agent (P=0.105). However, a high number of CD3 TILs was a favorable prognostic factor (P=0.008). Pearson's correlation analyses showed no association between cancer cell death rates in response to chemotherapeutic agents and subsets of TILs. Conclusions Cancer cell death rates in response to specific chemotherapeutic agents were not significantly associated with the distribution of TIL subsets. PMID:26819801

  19. OX40, PD-1 and CTLA-4 are selectively expressed on tumor-infiltrating T cells in head and neck cancer

    PubMed Central

    Montler, Ryan; Bell, R Bryan; Thalhofer, Colin; Leidner, Rom; Feng, Zipei; Fox, Bernard A; Cheng, Allen C; Bui, Tuan G; Tucker, Christopher; Hoen, Helena; Weinberg, Andrew

    2016-01-01

    The tumor microenvironment of squamous cell carcinoma of the head and neck (SCCHN) has been shown to be immune suppressive. Therefore, strategies aimed at overcoming this issue could have a positive therapeutic impact. Hence, we investigated the expression of the known immune-modulatory proteins OX40, programmed cell death protein 1 (PD-1) and cytotoxic T-lymphocyte-associated protein 4 (CTLA-4) in SCCHN on different T-cell subsets of tumor-infiltrating lymphocytes (TIL) to ascertain whether these proteins could potentially be targeted alone or in combination for future clinical trials. T cells from peripheral blood (PBL) and tumor were analyzed for the expression of OX40, PD-1 and CTLA-4 in 29 patients undergoing surgery. These proteins were all expressed significantly higher in T-cell subsets isolated from tumors compared with PBL of the same patient. OX40 expression was significantly greater in the TIL regulatory T-cell (Treg) population relative to conventional CD4 and CD8 TIL or the Treg isolated from PBL. PD-1 expression was increased in all T-cell subsets relative to PBL. CTLA-4 was also increased in all TIL subsets relative to blood, and similar to OX40, its highest level of expression was observed in the Treg TIL. The highest frequency of PD-1, CTLA-4 and OX40 triple-positive cells were found in the Treg population isolated from the tumor. We analyzed both human papilloma virus-positive and -negative patients and found similar levels and expression patterns of these two patient populations for all three proteins. These data suggest that there may be therapeutic advantages of targeting these pathways independently or in combination for patients with this disease. PMID:27195113

  20. TIGIT and PD-1 impair tumor antigen–specific CD8+ T cells in melanoma patients

    PubMed Central

    Chauvin, Joe-Marc; Pagliano, Ornella; Fourcade, Julien; Sun, Zhaojun; Wang, Hong; Sander, Cindy; Kirkwood, John M.; Chen, Tseng-hui Timothy; Maurer, Mark; Korman, Alan J.; Zarour, Hassane M.

    2015-01-01

    T cell Ig and ITIM domain (TIGIT) is an inhibitory receptor expressed by activated T cells, Tregs, and NK cells. Here, we determined that TIGIT is upregulated on tumor antigen–specific (TA-specific) CD8+ T cells and CD8+ tumor-infiltrating lymphocytes (TILs) from patients with melanoma, and these TIGIT-expressing CD8+ T cells often coexpress the inhibitory receptor PD-1. Moreover, CD8+ TILs from patients exhibited downregulation of the costimulatory molecule CD226, which competes with TIGIT for the same ligand, supporting a TIGIT/CD226 imbalance in metastatic melanoma. TIGIT marked early T cell activation and was further upregulated by T cells upon PD-1 blockade and in dysfunctional PD-1+TIM-3+ TA-specific CD8+ T cells. PD-1+TIGIT+, PD-1–TIGIT+, and PD-1+TIGIT– CD8+ TILs had similar functional capacities ex vivo, suggesting that TIGIT alone, or together with PD-1, is not indicative of T cell dysfunction. However, in the presence of TIGIT ligand–expressing cells, TIGIT and PD-1 blockade additively increased proliferation, cytokine production, and degranulation of both TA-specific CD8+ T cells and CD8+ TILs. Collectively, our results show that TIGIT and PD-1 regulate the expansion and function of TA-specific CD8+ T cells and CD8+ TILs in melanoma patients and suggest that dual TIGIT and PD-1 blockade should be further explored to elicit potent antitumor CD8+ T cell responses in patients with advanced melanoma. PMID:25866972

  1. Programmed death-1 ligand 1 and 2 are highly expressed in pleomorphic carcinomas of the lung: Comparison of sarcomatous and carcinomatous areas.

    PubMed

    Kim, Sehui; Kim, Moon-Young; Koh, Jaemoon; Go, Heounjeong; Lee, Dong Soo; Jeon, Yoon Kyung; Chung, Doo Hyun

    2015-11-01

    Pleomorphic carcinoma (PC) of the lung is a rare type of poorly differentiated non-small cell lung carcinoma (NSCLC) that belongs to sarcomatoid carcinoma (SC). It exhibits aggressive behaviour and resistance to chemotherapy and radiotherapy. Recently, immunotherapy targeting the programmed death-1 (PD-1)/PD ligand 1 (PD-L1) pathway has demonstrated favourable clinical outcomes in NSCLC. However, the expression patterns of PD-1-related molecules in pulmonary PC remain elusive. PD-L1 and PD-L2 expression was estimated in 41 cases of PC using immunohistochemistry. CD8(+) and PD-1(+) tumour-infiltrating lymphocytes (TILs) were also evaluated. PD-L1 and PD-L2 were highly expressed in pulmonary PCs (90.2% [37/41)]; 87.8% [36/41]). The amount of CD8(+) or PD-1(+) TILs and the ratio of PD-1(+)/CD8(+) TILs in PC were higher in males, smokers and older patients. PD-L1-positive PCs were infiltrated by higher numbers of CD8(+) TILs compared to PD-L1-negative cases (P=0.006). Of note, PD-L1 expression in pulmonary PCs was significantly higher in sarcomatous areas than in the carcinomatous portion (P=0.006). PC patients with a high ratio of PD-1(+)/CD8(+) TILs showed a shorter progression-free survival (P=0.036), whereas PD-L1 and PD-L2 expression had no prognostic implications. Our study demonstrates that pulmonary PCs very frequently express PD-L1 and PD-L2. Moreover, their expression is higher in sarcomatous cells than in carcinomatous areas. Thus, targeting the PD-1/PD-L1 pathway may represent a potential therapeutic candidate for this aggressive tumour.

  2. Impact of chemokine receptor CXCR3 on tumor-infiltrating lymphocyte recruitment associated with favorable prognosis in advanced gastric cancer.

    PubMed

    Li, Kai; Zhu, Zhengpeng; Luo, Jin; Fang, Jingyi; Zhou, Huanhuan; Hu, Min; Maskey, Ninu; Yang, Guifang

    2015-01-01

    Chemokine receptor CXCR3 has been proved to play an important role in tumorigenesis and tumor progression in many malignancies, but its precise efficacy on gastric cancer (GC) has not been evaluated yet. The present study was aimed to explore the correlation of chemokine receptor CXCR3 with tumor-infiltrating lymphocytes (TILs) and prognosis in advanced gastric cancer (GC). Expression of CXCR3 and CD4+, CD8+ TILs was conducted in 192 advanced GC specimens and 48 corresponding paracancerous tissues by immunohistochemical (IHC) analysis. CXCR3 expression in GC tissues was significantly higher than that in paracancerous tissues (P<0.001) and CD8+, CD4+ TILs infiltration increased with high CXCR3 expression (P=0.032 and P<0.001, respectively). Our study showed significantly lower CXCR3 expression in patients with greater tumor invasion depth and lymph node metastasis compared with patients with lesser tumor invasion depth and without lymph node metastasis (P=0.002 and P=0.001, respectively). Univariate analysis indicated that patients with high CXCR3 expression and high CD8+ TILs infiltration had longer overall survival (OS) (log-rank test, P<0.001 and P=0.002, respectively). Univariate and multivariate analyses indicated that CXCR3 expression was an independent prognostic factor for OS (P=0.002). The present study suggested that CXCR3 expression was upregulated in advanced GC and was associated with increased CD4+, CD8+ TILs infiltration and improved OS. Therefore, CXCR3 overexpression is implicated as a favorable prognostic biomarker in human advanced GC.

  3. Identification of a human melanoma antigen recognized by tumor-infiltrating lymphocytes associated with in vivo tumor rejection.

    PubMed Central

    Kawakami, Y; Eliyahu, S; Delgado, C H; Robbins, P F; Sakaguchi, K; Appella, E; Yannelli, J R; Adema, G J; Miki, T; Rosenberg, S A

    1994-01-01

    The cultured T-cell line TIL1200, established from the tumor-infiltrating lymphocytes (TILs) of a patient with advanced metastatic melanoma, recognized an antigen on most HLA-A2+ melanomas and on all HLA-A2+ cultured neonatal melanocytes in an HLA-A2 restricted manner but not on other types of tissues or cell lines tested. A cDNA encoding an antigen recognized by TIL1200 was isolated by screening an HLA-A2+ breast cancer cell line transfected with an expression cDNA library prepared from an HLA-A2+ melanoma cell line. The nucleotide and amino acid sequences of this cDNA were almost identical to the genes encoding glycoprotein gp100 or Pmel17 previously registered in the GenBank. Expression of this gene was restricted to melanoma and melanocyte cell lines and retina but was not expressed on other fresh or cultured normal tissues or other types of tumor tested. The cell line transfected with this cDNA also expressed antigen recognized by the melanoma-specific antibody HMB45 that bound to gp100. A synthetic 10-amino acid peptide derived from gp100 was recognized by TIL1200 in the context of HLA-A2.1. Since the administration of TIL1200 plus interleukin 2 resulted in regression of metastatic cancer in the autologous patient, gp100 is a possible tumor rejection antigen and may be useful for the development of immunotherapies for patients with melanoma. Images PMID:8022805

  4. A mechanism to explain the variations of tropopause and tropopause inversion layer in the Arctic region during a sudden stratospheric warming in 2009

    NASA Astrophysics Data System (ADS)

    Wang, Rui; Tomikawa, Yoshihiro; Nakamura, Takuji; Huang, Kaiming; Zhang, Shaodong; Zhang, Yehui; Yang, Huigen; Hu, Hongqiao

    2016-10-01

    The mechanism to explain the variations of tropopause and tropopause inversion layer (TIL) in the Arctic region during a sudden stratospheric warming (SSW) in 2009 was studied with the Modern-Era Retrospective analysis for Research and Applications reanalysis data and GPS/Constellation Observing system for Meteorology, Ionosphere, and Climate (COSMIC) temperature data. During the prominent SSW in 2009, the cyclonic system changed to the anticyclonic system due to the planetary wave with wave number 2 (wave2). The GPS/COSMIC temperature data showed that during the SSW in 2009, the tropopause height in the Arctic decreased accompanied with the tropopause temperature increase and the TIL enhancement. The variations of the tropopause and TIL were larger in higher latitudes. A static stability analysis showed that the variations of the tropopause and TIL were associated with the variations of the residual circulation and the static stability due to the SSW. Larger static stability appeared in the upper stratosphere and moved downward to the narrow region just above the tropopause. The descent of strong downward flow was faster in higher latitudes. The static stability tendency analysis showed that the strong downward residual flow induced the static stability change in the stratosphere and around the tropopause. The strong downwelling in the stratosphere was mainly induced by wave2, which led to the tropopause height and temperature changes due to the adiabatic heating. Around the tropopause, a pair of downwelling above the tropopause and upwelling below the tropopause due to wave2 contributed to the enhancement of static stability in the TIL immediately after the SSW.

  5. High numbers of granzyme B/CD8-positive tumour-infiltrating lymphocytes in nasopharyngeal carcinoma biopsies predict rapid fatal outcome in patients treated with curative intent.

    PubMed

    Oudejans, Joost J; Harijadi, Hari; Kummer, J Alain; Tan, I Bing; Bloemena, Elizabeth; Middeldorp, Jaap M; Bladergroen, Belinda; Dukers, Danny F; Vos, Wim; Meijer, Chris J L M

    2002-12-01

    This study determined whether tumour-infiltrating lymphocytes (TILs) in nasopharyngeal carcinomas (NPCs) include activated cytotoxic T lymphocytes (CTLs) and whether the numbers of activated CTLs in these biopsies are related to clinical outcome. Moreover, the study investigated whether the numbers of activated CTLs are associated with the expression of MHC class I proteins and the granzyme B antagonist PI-9 in the tumour cells. Forty-three Indonesian NPC patients (T(1-3), N(1-3), M(0)), who were treated with curative intent by radiotherapy only, were studied. Tumour-infiltrating activated CTLs were detected using antibodies against granzyme B, CD8, and CD56. Expression of MHC class I proteins and PI-9 was also determined by immunohistochemistry. Granzyme B-positive TILs were detected in all NPC biopsies. The presence of a high percentage (>25%) of granzyme B-positive TILs appeared to be a very strong predictor of a rapid fatal clinical outcome, independent of stage. Complete absence of MHC class I heavy chain expression in tumour cells was observed in 11 of 31 evaluable cases and low levels were observed in seven additional cases. No association between MHC class I expression and the numbers of granzyme B-positive TILs was observed. Expression of the granzyme B antagonist PI-9 in tumour cells was detected in three cases. It is concluded that the presence of many granzyme B-positive TILs in a selected group of Indonesian NPC patients is a strong and stage-independent marker for a rapid fatal clinical outcome.

  6. CD45RO+ Memory T Lymphocytes — a Candidate Marker for TNM-Immunoscore in Squamous Non–Small Cell Lung Cancer1

    PubMed Central

    Paulsen, Erna-Elise; Kilvaer, Thomas; Khanehkenari, Mehrdad Rakaee; Maurseth, Ramona Johansen; Al-Saad, Samer; Hald, Sigurd M.; Al-Shibli, Khalid; Andersen, Sigve; Richardsen, Elin; Busund, Lill-Tove; Bremnes, Roy; Donnem, Tom

    2015-01-01

    Tumor-infiltrating lymphocytes (TILs) are vital in limiting cancer progression and may supplement the TNM classification. CD45RO+ memory TILs show major prognostic impact in various malignancies but have not been extensively explored in non–small cell lung cancer (NSCLC). In this study, we aimed to evaluate their potential in a NSCLC TNM-Immunoscore. Tissue microarrays were constructed from tumor tissue samples from two cohorts including in total 536 patients (University Hospital of North Norway, n = 285; Nordland Hospital, n = 251) with primary resected stage I to IIIA NSCLC. The density of CD45RO+ and CD8+ TILs in tumor epithelial and stromal compartments of the tumors was evaluated by immunohistochemistry. In univariate analyses, intraepithelial CD45RO+ TIL density (T-CD45RO) was a significant prognostic factor for disease-specific survival (P = .007), limited to the squamous cell carcinoma (SCC) histology subgroup (P < .001), where it was significant in both cohorts (University Hospital of North Norway, P = .003; Nordland Hospital, P = .022). Combining T-CD45RO and stromal CD8+ TIL density (S-CD8) increased the prognostic impact in SCC (P < .001) and showed a significant impact within all pathological stages (I, P = .025; II, P < .001; III, P = .001). In the multivariate analysis, T-CD45RO was an independent positive prognostic factor for SCC (hazard ratio 2.65, 95% confidence interval 1.64-4.28, P < .001), and in combination with S-CD8, the prognostic impact increased vastly (high + high versus low + low: hazard ratio 6.50, 95% confidence interval 3.54-11.91, P < .001). In conclusion, T-CD45RO was an independent prognostic factor for SCC NSCLC. When combined with S-CD8, the prognostic impact increased and was significant within each pathological stage. We propose CD45RO as a candidate marker for TNM-Immunoscore in SCC NSCLC. PMID:26678911

  7. Prognostic impact of programmed cell death-1 (PD-1) and PD-ligand 1 (PD-L1) expression in cancer cells and tumor-infiltrating lymphocytes in ovarian high grade serous carcinoma

    PubMed Central

    Kulbe, Hagen; Sehouli, Jalid; Wienert, Stephan; Lindner, Judith; Budczies, Jan; Bockmayr, Michael; Dietel, Manfred; Denkert, Carsten; Braicu, Ioana; Jöhrens, Korinna

    2016-01-01

    Aims Antibodies targeting the checkpoint molecules programmed cell death 1 (PD-1) and its ligand PD-L1 are emerging cancer therapeutics. We systematically investigated PD-1 and PD-L1 expression patterns in the poor-prognosis tumor entity high-grade serous ovarian carcinoma. Methods PD-1 and PD-L1 protein expression was determined by immunohistochemistry on tissue microarrays from 215 primary cancers both in cancer cells and in tumor-infiltrating lymphocytes (TILs). mRNA expression was measured by quantitative reverse transcription PCR. An in silico validation of mRNA data was performed in The Cancer Genome Atlas (TCGA) dataset. Results PD-1 and PD-L1 expression in cancer cells, CD3+, PD-1+, and PD-L1+ TILs densities as well as PD-1 and PD-L1 mRNA levels were positive prognostic factors for progression-free (PFS) and overall survival (OS), with all factors being significant for PFS (p < 0.035 each), and most being significant for OS. Most factors also had prognostic value that was independent from age, stage, and residual tumor. Moreover, high PD-1+ TILs as well as PD-L1+ TILs densities added prognostic value to CD3+TILs (PD-1+: p = 0.002,; PD-L1+: p = 0.002). The significant positive prognostic impact of PD-1 and PD-L1 mRNA expression could be reproduced in the TCGA gene expression datasets (p = 0.02 and p < 0.0001, respectively). Conclusions Despite their reported immune-modulatory function, high PD-1 and PD-L1 levels are indicators of a favorable prognosis in ovarian cancer. Our data indicate that PD-1 and PD-L1 molecules are biologically relevant regulators of the immune response in high-grade serous ovarian carcinoma, which is an argument for the evaluation of immune checkpoint inhibiting drugs in this tumor entity. PMID:26625204

  8. The microenvironment in primary cutaneous melanoma with associated spontaneous tumor regression: evaluation for T-regulatory cells and the presence of an immunosuppressive microenvironment.

    PubMed

    Gray, Arielle; Grushchak, Solomiya; Mudaliar, Kumaran; Kliethermes, Stephanie; Carey, Kyle; Hutchens, Kelli A

    2017-04-01

    Spontaneous tumor regression, regression in the absence of therapeutic intervention, can be identified histologically in over 25% of primary cutaneous melanomas at initial diagnosis. A unique subset of T lymphocytes found in areas of regression can be histologically distinguished from tumor-infiltrating T lymphocytes (TIL) found in areas of tumor progression. We call this unique subset of T lymphocytes regression-associated T lymphocytes (RATs). The aim of this study is to determine the phenotype of lymphocytes and the density of specific cell types linked to immunosuppression in areas of tumor progression compared with areas of tumor regression. These specific cell types include T-regulatory cells (Tregs) and S100A9 cells. A total of 14 primary cutaneous melanomas with areas of progression and regression were used. Immunohistochemistry staining was used to identify CD4 cells, CD8 cells, Tregs, and S100A9 cells. Two independent observers manually counted three high-powered ×40 fields. There was no predominance of CD4 or CD8 T lymphocytes in either RATs or TIL. We identified a lower density of Tregs in RATs compared with TIL when using the FOXP3/CD4 Treg marker (P=0.04) and a marginal difference when using our second, confirmatory Treg marker, FOXP3/CD25 (P=0.11). We observed a lower density of S100A9 cells in RATs compared with TIL (P=0.002). There was an observable difference in the tumor microenvironments of RATs and TIL, with RATs having a significantly lower density of Tregs and S100A9 cells. We deduce that the absence of immunosuppression in areas of regression allows for a more robust immune response and thus effective eradication of tumor cells.

  9. TIGIT and PD-1 impair tumor antigen-specific CD8⁺ T cells in melanoma patients.

    PubMed

    Chauvin, Joe-Marc; Pagliano, Ornella; Fourcade, Julien; Sun, Zhaojun; Wang, Hong; Sander, Cindy; Kirkwood, John M; Chen, Tseng-hui Timothy; Maurer, Mark; Korman, Alan J; Zarour, Hassane M

    2015-05-01

    T cell Ig and ITIM domain (TIGIT) is an inhibitory receptor expressed by activated T cells, Tregs, and NK cells. Here, we determined that TIGIT is upregulated on tumor antigen-specific (TA-specific) CD8⁺ T cells and CD8⁺ tumor-infiltrating lymphocytes (TILs) from patients with melanoma, and these TIGIT-expressing CD8⁺ T cells often coexpress the inhibitory receptor PD-1. Moreover, CD8⁺ TILs from patients exhibited downregulation of the costimulatory molecule CD226, which competes with TIGIT for the same ligand, supporting a TIGIT/CD226 imbalance in metastatic melanoma. TIGIT marked early T cell activation and was further upregulated by T cells upon PD-1 blockade and in dysfunctional PD-1⁺TIM-3⁺ TA-specific CD8⁺ T cells. PD-1⁺TIGIT⁺, PD-1⁻TIGIT⁺, and PD-1⁺TIGIT⁻ CD8⁺ TILs had similar functional capacities ex vivo, suggesting that TIGIT alone, or together with PD-1, is not indicative of T cell dysfunction. However, in the presence of TIGIT ligand-expressing cells, TIGIT and PD-1 blockade additively increased proliferation, cytokine production, and degranulation of both TA-specific CD8⁺ T cells and CD8⁺ TILs. Collectively, our results show that TIGIT and PD-1 regulate the expansion and function of TA-specific CD8⁺ T cells and CD8⁺ TILs in melanoma patients and suggest that dual TIGIT and PD-1 blockade should be further explored to elicit potent antitumor CD8⁺ T cell responses in patients with advanced melanoma.

  10. Endonuclease G plays a role in immunoglobulin class switch DNA recombination by introducing double-strand breaks in switch regions.

    PubMed

    Zan, Hong; Zhang, Jinsong; Al-Qahtani, Ahmed; Pone, Egest J; White, Clayton A; Lee, Derrik; Yel, Leman; Mai, Thach; Casali, Paolo

    2011-01-01

    Immunoglobulin (Ig) class switch DNA recombination (CSR) is the crucial mechanism diversifying the biological effector functions of antibodies. Generation of double-strand DNA breaks (DSBs), particularly staggered DSBs, in switch (S) regions of the upstream and downstream CH genes involved in the specific recombination process is an absolute requirement for CSR. Staggered DSBs would be generated through deamination of dCs on opposite DNA strands by activation-induced cytidine deaminase (AID), subsequent dU deglycosylation by uracil DNA glycosylase (Ung) and abasic site nicking by apurinic/apyrimidic endonuclease. However, consistent with the findings that significant amounts of DSBs can be detected in the IgH locus in the absence of AID or Ung, we have shown in human and mouse B cells that AID generates staggered DSBs not only by cleaving intact double-strand DNA, but also by processing blunt DSB ends generated in an AID-independent fashion. How these AID-independent DSBs are generated is still unclear. It is possible that S region DNA may undergo AID-independent cleavage by structure-specific nucleases, such as endonuclease G (EndoG). EndoG is an abundant nuclease in eukaryotic cells. It cleaves single and double-strand DNA, primarily at dG/dC residues, the preferential sites of DSBs in S region DNA. We show here that EndoG can localize to the nucleus of B cells undergoing CSR and binds to S region DNA, as shown by specific chromatin immunoprecipitation assays. Using knockout EndoG(-/-) mice and EndoG(-/-) B cells, we found that EndoG deficiency resulted in a two-fold reduction in CSR in vivo and in vitro, as demonstrated by reduced cell surface IgG1, IgG2a, IgG3 and IgA, reduced secreted IgG1, reduced circle Iγ1-Cμ, Iγ3-Cμ, Iɛ-Cμ, Iα-Cμ transcripts, post-recombination Iμ-Cγ1, Iμ-Cγ3, Iμ-Cɛ and Iμ-Cα transcripts. In addition to reduced CSR, EndoG(-/-) mice showed a significantly altered spectrum of mutations in IgH J(H)-iEμ DNA. Impaired CSR in

  11. Charge sensing and real-time electron counting in quantum dots

    NASA Astrophysics Data System (ADS)

    Ihn, Thomas

    2012-02-01

    The charge state of a Coulomb blockaded quantum dot can be sensed at the single-electron level using charge sensors integrated on-chip [1]. Quantum point contacts or quantum dots have proven to work as reliable charge sensors. Coupling between the quantum dot's charge state and the sensor is provided by Coulomb interactions between electrons in the two systems. The technique is therefore independent of the material of quantum dot and sensor, and works for diverse systems such as GaAs [1] and graphene [2]. Different materials, such as InAs and GaAs have even been combined. Time-resolved charge sensing is of fundamental interest for studying the statistical properties of charge flow, like the full counting statistics. Single-shot charge and spin read-out schemes are also needed for the measurement of qubits. Some of our recent work has focused on the exploration and optimization of the read-out bandwidth limits. Towards this goal, we learned how to influence the dot-detector coupling [3], and how to employ high-frequency techniques in the range of a few hundred megahertz [4], or even up to a few gigahertz for improved performance. New experiments using the slower conventional sensing schemes have allowed us to explore non-equilibrium statistics and the fluctuation theorem. In our measurements, rare events can be detected, where electrons flow against the applied source-drain bias direction thereby consuming entropy in agreement with theoretical predictions. [4pt] [1] T. Ihn, S. Gustavsson, U. Gasser, R. Leturcq, I. Shorubalko, and K. Ensslin, Physica E: Low-dimensional Systems and Nanostructures 42, 803-808 (2010).[0pt] [2] J. G"uttinger, J. Seif, C. Stampfer, A. Capelli, K. Ensslin, and T. Ihn, Phys. Rev. B 83, 165445 (2011).[0pt] [3] C. R"ossler, B. K"ung, S. Dr"oscher, T. Choi, T. Ihn, K. Ensslin, and M. Beck, Appl. Phys. Lett. 97, 152109 (2010).[0pt] [4] T. M"uller, B. K"ung, S. Hellm"uller, P. Studerus, K. Ensslin, T. Ihn, M. Reinwald, and W. Wegscheider, Appl

  12. Experimental demonstration of coherent beam combining over a 7 km propagation path.

    PubMed

    Weyrauch, Thomas; Vorontsov, Mikhail A; Carhart, Gary W; Beresnev, Leonid A; Rostov, Andrey P; Polnau, Ernst E; Liu, Jony Jiang

    2011-11-15

    We demonstrate coherent combining (phase locking) of seven laser beams emerging from an adaptive fiber-collimator array over a 7 km atmospheric propagation path using a target-in-the-loop (TIL) setting. Adaptive control of the piston and the tip and tilt wavefront phase at each fiber-collimator subaperture resulted in automatic focusing of the combined beam onto an unresolved retroreflector target (corner cube) with precompensation of quasi-static and atmospheric turbulence-induced phase aberrations. Both phase locking (piston) and tip-tilt control were performed by maximizing the target-return optical power using iterative stochastic parallel gradient descent (SPGD) techniques. The performance of TIL coherent beam combining and atmospheric mitigation was significantly increased by using an SPGD control variation that accounts for the round-trip propagation delay (delayed SPGD).

  13. Target-in-the-loop remote sensing of laser beam and atmospheric turbulence characteristics.

    PubMed

    Vorontsov, Mikhail A; Lachinova, Svetlana L; Majumdar, Arun K

    2016-07-01

    A new target-in-the-loop (TIL) atmospheric sensing concept for in situ remote measurements of major laser beam characteristics and atmospheric turbulence parameters is proposed and analyzed numerically. The technique is based on utilization of an integral relationship between complex amplitudes of the counterpropagating optical waves known as overlapping integral or interference metric, whose value is preserved along the propagation path. It is shown that the interference metric can be directly measured using the proposed TIL sensing system composed of a single-mode fiber-based optical transceiver and a remotely located retro-target. The measured signal allows retrieval of key beam and atmospheric turbulence characteristics including scintillation index and the path-integrated refractive index structure parameter.

  14. Expression of immune checkpoints in T cells of esophageal cancer patients

    PubMed Central

    Xie, Jinhua; Wang, Ji; Cheng, Shouliang; Zheng, Liangfeng; Ji, Feiyue; Yang, Lin; Zhang, Yan; Ji, Haoming

    2016-01-01

    Inhibition of immune checkpoint proteins (checkpoints) has become a promising anti-esophageal cancer strategy. We here tested expressions of immune checkpoints in human esophageal cancers. Our results showed the expressions of many immune checkpoints, including CD28, CD27, CD137L, programmed death 1 (PD-1), T cell immunoglobulin mucin-3 (TIM-3), T cell Ig and ITIM domain (TIGIT), CD160, cytotoxic T lymphocyte antigen 4 (CTLA-4), CD200, CD137 and CD158, were dysregulated in peripheral T cells of esophageal cancer patients. Further, the expressions of PD-1, TIM-3 and TIGIT were upregulated in tumor infiltrating lymphocytes (TILs), which might be associated with TILs exhaustion. Meanwhile, the expressions of PD-1 and TIM-3 on CD4+ T cells were closely associated with clinic pathological features of esophageal cancer patients. These results indicate that co-inhibitory receptors PD-1, TIM-3 and TIGIT may be potential therapeutic oncotargets for esophageal cancer. PMID:27577071

  15. Habitat fragmentation influences gene structure and gene differentiation among the Loxoblemmus aomoriensis populations in the Thousand Island Lake.

    PubMed

    Lv, Kun; Zhou, Jing; Gu, Jian-Qiang; Zhou, Guo-Xing; Wang, Wei; Xu, Zhi-Hong

    2017-02-16

    Thousand Island Lake (TIL) is a fragmented landscape consisting of more than 1000 land-bridge islands isolated during reservoir formation. To evaluate the effects of fragmentation and island attributes on insect populations, we examined the genetic structure of Loxoblemmus aomoriensis, a species of cricket widely distributed in TIL, and compared genetic diversity between islands samples. Population genetic analyses was conducted based on mitochondrial DNA haplotype frequencies of 10 sample islands. By comparing three island attributes with population genetic diversity reveals that island area influenced population genetic diversity (r(2 )=( )0.5094, p = 0.00204). Using Pairwise Fst values, we also found that long-distance isolation increased the genetic differentiation, while short-distance isolation can be offset by dispersal. These results indicate that fragmentation can impact populations on a genetic level.

  16. Exploiting natural anti-tumor immunity for metastatic renal cell carcinoma

    PubMed Central

    Murphy, Katherine A; James, Britnie R; Guan, Yue; Torry, Donald S; Wilber, Andrew; Griffith, Thomas S

    2015-01-01

    Clinical observations of spontaneous disease regression in some renal cell carcinoma (RCC) patients implicate a role for tumor immunity in controlling this disease. Puzzling, however, are findings that high levels of tumor infiltrating lymphocytes (TIL) are common to RCC. Despite expression of activation markers by TILs, functional impairment of innate and adaptive immune cells has been consistently demonstrated contributing to the failure of the immune system to control RCC. Immunotherapy can overcome the immunosuppressive effects of the tumor and provide an opportunity for long-term disease free survival. Unfortunately, complete response rates remain sub-optimal indicating the effectiveness of immunotherapy remains limited by tumor-specific factors and/or cell types that inhibit antitumor immune responses. Here we discuss immunotherapies and the function of multiple immune system components to achieve an effective response. Understanding these complex interactions is essential to rationally develop novel therapies capable of renewing the immune system's ability to respond to these tumors. PMID:25996049

  17. No Further Action Decision Under CERCLA, Fort Devens Study Area 28 Waste Explosives Detonation Range (Training Area 14)

    DTIC Science & Technology

    1994-01-01

    Environmental Response, Compensation and Lability Act ( CERCLA ) as amended by the Superfund Amendments and Reauthorization Act. An Enhanced Preliminary...til3i NO FURT ACTION DECISION UNDER CERCLA FORT DEVENS STUDY AREA 28 WASTE EXPLOSIVES DETONATION RANGE (TRAINING AREA 14) DATA ITEM A009...UNDER CERCLA STUDY AREA 28 WASTE EXPLOSIVES DETONATION RANGE (TRAINING AREA 14) FORT DEVENS, MASSACHUSETTS Prepared for: U.S. Army Environmental

  18. Detection and Characterization of a Novel Subset of CD8+CD57+ T-cells in Metastatic Melanoma with an Incompletely-Differentiated Phenotype

    PubMed Central

    Wu, Richard C.; Liu, Shujuan; Chacon, Jessica A.; Wu, Sheng; Li, Yufeng; Sukhumalchandra, Pariya; Murray, James L.; Molldrem, Jeffrey J.; Hwu, Patrick; Pircher, Hanspeter; Lizée, Gregory; Radvanyi, Laszlo G.

    2012-01-01

    PURPOSE Tumor-specific T-cells are frequently induced naturally in melanoma patients and infiltrate tumors. It is enigmatic why these patients fail to experience tumor regression. Given that CD8+ T cells mediate antigen-specific killing of tumor cells, the focus of this study was to identify alterations in the differentiation of CD8+ residing at the tumor site, with emphasis on a population expressing CD57, a marker for terminal differentiation. EXPERIMENTAL DESIGN We performed flow cytometric analysis of CD8+ tumor-infiltrating lymphocytes (TIL) isolated from 44 resected melanoma metastases using known T-cell differentiation markers. For comparison, PBMC were isolated from matched melanoma patients. We sorted different CD8+ subsets found in TIL and determined their effector functions. In addition, we performed Vβ spectratyping of T-cell receptors to determine lineage relationship between the CD8+ TIL subsets. RESULTS The majority of CD8+ TIL were in the early effector-memory stage of differentiation. A significant population consisted of an oligoclonal subset of cells co-expressing early effector-memory markers and end-stage CTL marker, CD57, yet having low to absent perforin expression. These cells could be induced to proliferate, produce a high level of IFN-γ, and differentiate into CD27−CD57+, perforinhigh mature CTL in vitro. Addition of TGF-β1 prevented this further differentiation. CONCLUSIONS Our studies identified a novel subset of incompletely differentiated CD8+ CTL co-expressing early effector-memory and late CTL markers. This population resembles that found by in patients with uncontrolled chronic viral infections. TGF-β1, frequently produced by melanoma tumors, may be a key cytokine inhibiting the further maturation of this subset. PMID:22307139

  19. Targeting PD-1 and Tim-3 Pathways to Reverse CD8 T-Cell Exhaustion and Enhance Ex Vivo T-Cell Responses to Autologous Dendritic/Tumor Vaccines.

    PubMed

    Liu, Jingwei; Zhang, Shurong; Hu, Yuefeng; Yang, Zhaomin; Li, Jingpo; Liu, Xuesong; Deng, Lijuan; Wang, Yue; Zhang, Xiaoyan; Jiang, Ting; Lu, Xu

    2016-05-01

    The paradoxical coexistence of spontaneous tumor antigen-specific immune response with progressive disease in cancer patients need to dissect the molecular pathways involved in tumor-induced T-cell dysfunction or exhaustion. Programmed cell death 1 (PD-1) has been identified as a marker of exhausted T cells in chronic disease states, and blockade of PD-1-PD-L1 interactions has been shown to partially restore T-cell function. We have found that T-cell immunoglobulin mucin (Tim) 3 is expressed on CD8+ tumor-infiltrating lymphocytes (TILs) isolated from patients with colorectal cancer. All T-cell immunoglobulin mucin 3 (Tim-3+) TILs coexpress PD-1, and Tim-3+ PD-1+ CD8+ TILs represent the predominant fraction of Tcells infiltrating tumors. Tim-3+PD-1+ CD8+ TILs exhibit the most severe exhausted phenotype as defined by failure to produce cytokines, such as interferon-γ, tumor necrosis factor-α, and interleukin-2. We further find that combined targeting of the Tim-3 and PD-1 pathways increased the frequencies of not only interferon-γ and tumor necrosis factor-α but also frequencies of proliferating tumor antigen-specific CD8+ T cells than targeting either pathway alone. A concomitant decrease in regulatory T cells and enhanced killing in a cytotoxicity assay was observed. Collectively, our findings support the use of Tim-3-Tim-3L blockade together with PD-1-PD-L1 blockade to reverse tumor-induced T-cell exhaustion/dysfunction in patients with colorectal cancer.

  20. United States Air Force Statistical Digest 1948, Third Annual Edition, Volume 1

    DTIC Science & Technology

    1948-12-31

    The attached material, described below, is forwarded for security and policy review in accordance with AFI 35-101, Chapter 15: TITLE: USAF Summaries...21 jail 2011 Office of the Secretary MEMORANDUM fOR AFIHO FROM: SAFIPA (Security and Policy Review) SUBJECT: Public Release Coordination (PAIRS...SAFJPA (Security and Policy Review) to objection _No objection, subject to recommendation DEV ALEE PRlDGEN-GA TIlSON Security Review Specialist

  1. Multiple-Input Transfer Function Model of Heat Transfer from Square Slab Floors

    DTIC Science & Technology

    1990-01-01

    this study. Further work to develop a definition of the network parameters based on characteristic length could expand the use of the model to non...Validation 57 6 NETWORK PARAMETERS BASED ON CHARACTERISTIC LENGTH .... 78 7 UTILIZATION OF TIlE GTF MODEL FOR ENERGY ANALYSIS ....... 87 8 CONCLUSIONS...BASIC PROGRAM GTF 96 APPENDIX C: TRUBASIC PROGRAM QCALC 108 DISTRIBUTION vi LIST OF FIGURES Figure Number Page 1 7-Node Network Model

  2. Analysis of the hippo transducers TAZ and YAP in cervical cancer and its microenvironment

    PubMed Central

    Buglioni, Simonetta; Vici, Patrizia; Sergi, Domenico; Pizzuti, Laura; Di Lauro, Luigi; Antoniani, Barbara; Sperati, Francesca; Terrenato, Irene; Carosi, Mariantonia; Gamucci, Teresa; Vincenzoni, Cristina; Mariani, Luciano; Vizza, Enrico; Venuti, Aldo; Sanguineti, Giuseppe; Gadducci, Angiolo; Barba, Maddalena; Natoli, Clara; Vitale, Ilio; Mottolese, Marcella; De Maria, Ruggero; Maugeri-Saccà, Marcello

    2016-01-01

    ABSTRACT Hippo is a tumor-suppressor pathway that negatively regulates the oncoproteins TAZ and YAP. Moreover, Hippo affects the biology of a variety of non-neoplastic cells in the tumor microenvironment, even including immune cells. We herein assessed the predictive role of TAZ and YAP, assessed by immunohistochemistry, in 50 cervical cancer patients prevalently treated with neoadjuvant chemotherapy. Tumors were classified as positive or negative according to the percentage of tumor-expressing cells and cellular localization. TAZ/YAP were also evaluated in non-neoplastic cells, namely endothelial cells, non-lymphocytic stromal cells and tumor-infiltrating lymphocytes (TILs). TAZ expression in cancer cells (TAZpos) was associated with a reduced pathological complete response (pCR) rate (p = 0.041). Conversely, the expression of TAZ and YAP in TILs (TAZTIL+ and YAPTIL+) seemed to be associated with increased pCRs (p = 0.083 and p = 0.018, respectively). When testing the predictive significance of the concomitant expression of TAZ in cancer cells and its absence in TILs (TAZpos/TAZTIL-), patients with TAZpos/TAZTIL- showed lower pCR rate (p = 0.001), as confirmed in multivariate analysis (TAZpos/TAZTIL-: OR 8.67, 95% CI: 2.31–32.52, p = 0.001). Sensitivity analysis carried out in the 41 patients treated with neoadjuvant chemotherapy yielded comparable results (TAZpos/TAZTIL-: OR 11.0, 95% CI: 2.42–49.91, p = 0.002). Internal validation carried out with two different procedures confirmed the robustness of this model. Overall, we found evidence on the association between TAZ expression in cervical cancer cells and reduced pCR rate. Conversely, the expression of the Hippo transducers in TILs may predict increased treatment efficacy, possibly mirroring the activation of a non-canonical Hippo/MST pathway necessary for T-cells activation and survival. PMID:27471633

  3. The prognostic value of systemic and local inflammation in patients with laryngeal squamous cell carcinoma

    PubMed Central

    Wang, Jie; Wang, Shengzi; Song, Xinmao; Zeng, Wenjiao; Wang, Shuyi; Chen, Fu; Ding, Hao

    2016-01-01

    Background Cancer-related systemic inflammation has been demonstrated to be associated with poor outcome in multiple types of cancers. Meanwhile, the local inflammation, which is characterized by dense intratumoral immune infiltrate, is a favorable predictor of survival outcome. Purpose To evaluate the role of systemic and local inflammation in predicting outcome in patients with laryngeal squamous cell carcinoma. Patients and methods In this retrospective study, 120 patients who had undergone postoperative radiotherapy were enrolled. Neutrophil-to-lymphocyte ratio (NLR) and platelet-to-lymphocyte ratio (PLR), as calculated from pretreatment whole blood counts, were used to indicate systemic inflammation. The optimal cutoff values of NLR and PLR were determined using receiver operating characteristic curve analysis. Tumor infiltrating lymphocytes (TILs) density, as assessed by pathologist review of hematoxylin and eosin-stained slides, was used to represent local inflammation. Overall survival (OS) and recurrence-free survival (RFS) were assessed using the Kaplan–Meier method and multivariate Cox regression analysis. Results The best cutoff was 2.79 for NLR and 112 for PLR. Kaplan–Meier analysis revealed that high NLR, high PLR, and low TILs density were significantly correlated with inferior OS and RFS, respectively (all P<0.05). The Cox proportional multivariate hazard model showed that a high pretreatment PLR and a low TILs density were both independently correlated with poor OS and RFS, respectively (all P<0.05). Conclusion Markers of systemic and local inflammation, especially PLR and TILs density, are reliable prognostic factors in patients with laryngeal squamous cell carcinoma. PMID:27920556

  4. OPSATCOM Field Measurements. Volume 1

    DTIC Science & Technology

    1976-06-01

    1-2 S2 FORMULATION OF TIlE EXPERIMENTAL MODEL ........ 2-i 3 EXPERIMENTAL METHODS AND APPARATUS ........... 3-I 3.1 Methods Utilized...lfactors for investigating the optical clharactcr ol’ the seab, For the earliest work, which dates back to 1885, only photographic methods were...I 2-3/2-4 blank I I SECTION 3 I EXPERIMENTAL METHODS AND APPARATUS 3.1 METHODS UTILIZED In general, the atmospheric radiance pattern of’ an underwater

  5. Spacecraft Charging Standard Report.

    DTIC Science & Technology

    1980-09-30

    30.1.3 Backscattering. The deflection of particles by scattering processes in matter such that particles emerge through the same planar surface as they...with filters to provide protection against high frequency (up to 100 MHz), large | amplitude (TBD amperes), fast rise time (< 10 nanoseconds ) pulses of...A discharne con- ment is comprehensive, le vacuum - 100 nsec to I lisec pulse cepts acceptable, tIL -STD-I541, 461. edlations width fiber optics

  6. Net Shape Technology in Aerospace Structures. Volume 1.

    DTIC Science & Technology

    1986-11-01

    maintained at operating temperature b\\ induct ion, rc,,istancc, or gas-fired infrared heating . Induction heating is most Colmon for t it:1 i tIl forgings and...forging titanium at higher than .1750 0 F. 8. Research and dcclopment on die heating systems employing combinations of induction , resistance, and gas...methods being c aluated includc usc of graphite reinforc:ement for resistance hcating a portable induction heating unit dec loped at N -\\SA-Langle

  7. An Analysis of Transmittances Measured through Battlefield Dust Clouds

    DTIC Science & Technology

    1980-02-01

    110 ’N I-000000000 ~~c 0 00 00 Best Available Copy .99 .99 Ř. U ""•i~ lt)Il*) • , -4 I * I 49 9,.99 ,*.t~*Il . .99. . , ,.,’, ., . . * ,t • , , ,j It...599,August 1978. 36.Ducan Lui D. "trtosheicWind Shear Coptdfrom Satellite Thermal SouderMeaureent,"ECOM .8800. September 1978. 37. Tayor F. P.MoanP

  8. The Coast Artillery Journal. Volume 64, Number 6, June 1926

    DTIC Science & Technology

    1926-06-01

    continue its mission of covering the left flank. The 69th Reserve Division was directed upon Solre -Ie-Chateau, with a ,-iew to moving toward Beaumont or...CTURED BY BARTO~ l\\lAl\\’UF ACTURING CO;\\IPANY TilE AWE PAIn. A ~hort time a~o the aide to onc of onT g(’lwral OffiCf’TlS, driving at night, was

  9. Dielectric Constant and Loss Data, Part 3

    DTIC Science & Technology

    1977-05-01

    to the general public, including foreign nations. This technical report has been reviewed and is approved. JOHN C. OLSON Project Engineer FOR TIlE...PERFORIIINO ORGANIZATION NA E AND ADDRESS 10. ROORAM ELEMENT PROJECT , TASKC It. CONTROLLING OFFICE NAME AND ADDRESS Laboratory for Insulation Research...Massachusetts, by W. B. Westphal. This work was performed between I July 1974 and 31 December 1976 under Contract F33615-75-C-5020, Project No. 7371, Task No

  10. US EPA, Pesticide Product Label, , 03/23/1983

    EPA Pesticide Factsheets

    2011-04-14

    ... flor/eflll9 tn·fH., 1 dnd (>'."I'!'(J~f'L'''<' f(lund ~o be inf(,~t."d ,·:ith 1,(,,·t5 1i"t(·rj iii til" fr-ll',;Il'l t,Ll". lJi lute DIJRSEf,ti ~'):: Ii th I".! ((r dl'.wni Il'J to di fee. ...

  11. Task Force Delay Study, Miami International Airport.

    DTIC Science & Technology

    1981-07-01

    13 . .Pe of Pepo., ond P.,iod Co-e,ed 12. Spo’s-: Agency No-.. and Ad. ness U.S. Department of Transportation Fejr ! Aviation Administration...W, 66 U : ~ Ssz tz - c~7j ~~nz Ll ss til-= - -j E E~ -- n -3 .)- m -Q - v c. ~c E c- ’Z -. E- --. It C. ,: -E mo ;. .- - -VC c-&-,J.tv --- -t -. be

  12. Thermal Conductivity of Soils

    DTIC Science & Technology

    1977-07-01

    Rapport sendt til NTNF. Geologisk Institutt, NTH, Trondheim 1961, p. 29 f.f.. R. Selmer-Olsen: IngeniOrgeologi. Del 1. Generell geologi. Tapir forlag...Ingeniorgeologi, Del 1, Generell geologi, Tapir forlag, Troniheim 1971, p. 175. 5. H. Sveian2 Norske jordarters kvartsinnhold. Hoveioppgave i Ingeni~orgeologi...I 16. R. Selmer-Olsen: Ingeni~rgeologi. Del I. Generell geologi. Tapir forlag. Tronheim 1971, p. 8. 17. R. Selmer-Olsen: Ingeni~rgeologi. Del I

  13. Surface Observation Climatic Summaries (SOCS) for Hanau AAF, Germany

    DTIC Science & Technology

    1992-03-01

    oI L ...............- A V EtTIONH NAI6s mADAM AAF DL FIELD ELI’Y: 363 Fr LATITIUDE/LECSITUDEs 50 10 N 008 58 E STATIOC N C: 106420...r AVAILAULE FOR WMER REPO~nTil SITES. AIRIIEAR SEITIG. 3INCER OF CURY (80), TABLES GIVE Hk, W!nffARDA DEVIATIONS, AND SINTON VIOUM.YN IF INESS( MN CW

  14. Serum ferritin level changes in children with sickle cell disease on chronic blood transfusion are nonlinear and are associated with iron load and liver injury

    PubMed Central

    Abboud, Miguel R.; Paley, Carole; Olivieri, Nancy; Kirby-Allen, Melanie; Vichinsky, Elliott; Casella, James F.; Alvarez, Ofelia A.; Barredo, Julio C.; Lee, Margaret T.; Iyer, Rathi V.; Kutlar, Abdullah; McKie, Kathleen M.; McKie, Virgil; Odo, Nadine; Gee, Beatrice; Kwiatkowski, Janet L.; Woods, Gerald M.; Coates, Thomas; Wang, Winfred; Adams, Robert J.

    2009-01-01

    Chronic blood transfusion is increasingly indicated in patients with sickle cell disease. Measuring resulting iron overload remains a challenge. Children without viral hepatitis enrolled in 2 trials for stroke prevention were examined for iron overload (STOP and STOP2; n = 271). Most received desferrioxamine chelation. Serum ferritin (SF) changes appeared nonlinear compared with prechelation estimated transfusion iron load (TIL) or with liver iron concentrations (LICs). Averaged correlation coefficient between SF and TIL (patients/observations, 26 of 164) was r = 0.70; between SF and LIC (patients/observations, 33 of 47) was r = 0.55. In mixed models, SF was associated with LIC (P = .006), alanine transaminase (P = .025), and weight (P = .026). Most patients with SF between 750 and 1500 ng/mL had a TIL between 25 and 100 mg/kg (72.8% ± 5.9%; patients/observations, 24 of 50) or an LIC between 2.5 and 10 mg/g dry liver weight (75% ± 0%; patients/observations, 8 of 9). Most patients with SF of 3000 ng/mL or greater had a TIL of 100 mg/kg or greater (95.3% ± 6.7%; patients/observations, 7 of 16) or an LIC of 10 mg/g dry liver weight or greater (87.7% ± 4.3%; patients/observations, 11 of 18). Although SF changes are nonlinear, levels less than 1500 ng/mL indicated mostly acceptable iron overload; levels of 3000 ng/mL or greater were specific for significant iron overload and were associated with liver injury. However, to determine accurately iron overload in patients with intermediately elevated SF levels, other methods are required. These trials are registered at www.clinicaltrials.gov as #NCT00000592 and #NCT00006182. PMID:19721013

  15. US EPA, Pesticide Product Label, NOVA CONTACT SPRAY ...

    EPA Pesticide Factsheets

    2011-04-21

    I , I " ! '.'f '( i .' l' \\. : ' t ' ~. :' I .1: t . ( . \\ J r )I i It, () I H'! J Il {f ve n t II d t (} f (, ''/ tIl! (l. S l' i! "I; t q ~ p r ; V"I)t" C(),'PI ,.Ii f(lt)d ild!llllll;tj :;\\11 1(1':"', diHi l'(),·1 ...

  16. Potential Prognostic Markers for Human Prostate Cancer

    DTIC Science & Technology

    2001-03-01

    such as proliferation, differentiation and assay [15]. TGF[3 may be produced by metastatic motility. Upregulation of growth factor synthesis or prostate...via mod- ronment include neuroendocrine cells. These cells ulation of tumor cell receptor expression. Control of can secrete bombesin, a gastrin ...controls, indicating that addition ofT015 accelerates tumor expansion. TIl15 synthesis triggered a dramatic increase in cell motility, boosting serum

  17. Immunophenotyping of patients with oral squamous cell carcinoma in peripheral blood and associated tumor tissue.

    PubMed

    Grimm, Martin; Feyen, Oliver; Hofmann, Heiko; Teriete, Peter; Biegner, Thorsten; Munz, Adelheid; Reinert, Siegmar

    2016-03-01

    The immune system is important for elimination of cancer cells. Tumors including oral squamous cell carcinoma (OSCC) are capable of escaping detection by host immune cells through apoptotic depletion of tumor-infiltrating lymphocytes (TILs). Circulating peripheral blood lymphocytes (PBLs) and corresponding TILs of tumor specimen were evaluated before and after curative tumor resection (n = 30) compared with PBLs of controls (n = 87). PBLs were characterized for the total number of T cells (CD3(+)), T helper cells (Th, CD3(+)/CD4(+)), regulatory T cells (Treg, CD4(+)/CD25(+)/CD127(low)), cytotoxic T cells (Tc, CD3(+)/CD8(+)), activated T cells (CD3(+)/HLA-DR(+)), and natural killer (NK) cells (CD3(-)/CD16(+)/CD56(+)). In tumor tissue, the prevalence of CD3(+), CD4(+), and CD8(+) TILs was assessed using immunohistochemistry, whereas the incidence of apoptosis was assessed using terminal deoxynucleotidyl transferase deoxyuridinetriphosphate nick-end labeling (TUNEL) assay. In PBLs of pretreated OSCC patients, a highly significant decrease in total number of T cells (p = 0.0001), Th cells (p < 0.0001), Treg cells (p < 0.0001), Tc cells (p < 0.0001), and NK cells (p = 0.0037) were found compared with controls. Decreased PBLs of OSCC patients were correlated with decreased numbers of corresponding TILs, which were associated with increased detection of apoptosis in the tumor tissue. Compared with the controls, the total number of T cells remained unchanged after surgery but the total number of NK cells significantly increased. Standardized immunophenotyping of OSCC may help to identify patients likely to benefit from cancer immunotherapy strategies and/or chemoradiation. Finally, future attempts to enhance an effective tumor-reactive immune response by immunotherapy or vaccination should be made by promoting tumor-specific Th and/or Tc cell/NK cell responses.

  18. Regulation of the Prostate Cancer Tumor Microenvironment

    DTIC Science & Technology

    2012-04-01

    infiltrating macrophage lineage in the absence of MyD88. We are in the process of understanding the activation of signaling pathways , local and...development, growth, and metastasis is unclear. We are interested in understanding the mechanisms for development of TILs and how they modulate prostate...chromatin component HMG-B1. Activation of these receptors leads to induction of multiple inflammatory pathways , including nuclear factor-kappa B (NF

  19. US EPA, Pesticide Product Label, EMULSAMINE BK WOODY ...

    EPA Pesticide Factsheets

    2011-04-13

    ... T ".., 11I1'Ih,l' ('I" tit' u'>f'd til a", I,""'f' 01 )"'d' lIt .. t fl'lp t">dSP vI d,' "It'",," Un!lI s,"a, Pdddlt"~ '" ,t1(olit>Cla l.j'l(l~"f'f;·"""dfit·Q'(·LJ·'~ 'It' li< .• , I .. ", ...

  20. AGARD/SMP Review Damage Tolerance for Engine Structures. 2. Defects and Quantitative Materials Behaviour

    DTIC Science & Technology

    1988-10-01

    temperature range, moderate strength - two broad lines of developments have been conducted : . alloys with high specific strength for medium... temperatures applications (^♦00 to ^500C) : Ti 6.2.4.6. , Til? , IMI 550 , ... . alloys for high temperature applications : Ti 6.2.4.2.3. , IMI 685 , IMI 829...cation of Titanium alloys established upon the relative stability of the low temperature (hep) and high temperature (bec) phase : this classifi- cation

  1. Tumor-infiltrating Tim-3(+) T cells proliferate avidly except when PD-1 is co-expressed: Evidence for intracellular cross talk.

    PubMed

    Li, Jing; Shayan, Gulidanna; Avery, Lyndsay; Jie, Hyun-Bae; Gildener-Leapman, Neil; Schmitt, Nicole; Lu, Bin Feng; Kane, Lawrence P; Ferris, Robert L

    2016-01-01

    Programmed Death 1 (PD-1) and T cell Ig and mucin domain-3 protein (Tim-3) are immune checkpoint receptors highly expressed on tumor infiltrating T lymphocytes (TIL). PD-1 inhibits T cell activation and type-1 T cell responses, while Tim-3 is proposed to mark more extensively exhausted cells, although the mechanisms underlying Tim-3 function are not clear. Trials of anti-PD-1 therapy have identified a large subset of non-responder patients, likely due to expression of alternative checkpoint molecules like Tim-3. We investigated the phenotypic and functional characteristics of T cells with differential expression of PD-1 (high/low) and Tim-3 (positive/negative), using TIL directly isolated from head and neck squamous cell carcinomas (HNSCC). Unexpectedly, we found that expression of Tim-3 alone does not necessarily mark TIL as dysfunctional/exhausted. In Tim-3-TIL, PD-1 levels correlate with T cell dysfunction, with a PD-1(low/intermed) phenotype identifying recently activated and still functional cells, whereas PD-1(hi)Tim-3(-) T cells are actually exhausted. Nonetheless, PD-1(intermed) cells are still potently suppressed by PD-L1. PD-1 expression was associated with reduced phosphorylation of ribosomal protein S6 (pS6), whereas Tim-3 expression was associated with increased pS6. Using a novel mouse model for inducible Tim-3 expression, we confirmed that expression of Tim-3 does not necessarily render T cells refractory to further activation. These results suggest the existence of PD-1 and Tim-3 crosstalk in regulating antitumor T cell responses, with important implications for anti-PD-1 immunotherapy.

  2. Concordance of immune checkpoints within tumor immune contexture and their prognostic significance in gastric cancer.

    PubMed

    Dai, Congqi; Geng, Ruixuan; Wang, Chenchen; Wong, Angela; Qing, Min; Hu, Jianjun; Sun, Yu; Lo, A W I; Li, Jin

    2016-12-01

    Checkpoint blockade therapy has emerged as a novel approach for cancer immunotherapy in several malignancies. However, patient prognosis and disease progression relevant to immune checkpoints in gastric tumor microenvironment are not defined. This study aims to investigate the expression and prognostic significance of immune checkpoints within gastric cancer. In the study, a cohort of 398 cancer tissues from stage I to IV gastric cancer patients were assessed for programmed cell death 1 ligand 1 (PD-L1) expression and tumor-infiltrating lymphocyte (TIL) infiltration using immunohistochemistry to ascertain their survival correlation. The data revealed that higher TIL density correlated with less risk of disease progression, and exhibited survival benefits in gastric cancer patients, and PD-L1 positivity showed a significant association with the presence of high TIL infiltration. Furthermore, real-time quantitative polymerase chain reaction was performed to detect expression of multiple immune checkpoints with the relation to clinical outcome in 139 samples randomly selected from the same cohort, and higher messenger RNA levels of most immune checkpoints were associated with favorable outcome, while consistently showing a positive correlation with interferon gamma levels. In situ hybridization was used to determine the localization of Epstein-Barr virus (EBV) in 97 specimens, and showed EBV-positive gastric cancer samples correlated with PD-L1 expression and increased TIL density. These results suggest that induction of immune checkpoint within gastric cancer patients reflects a high immune infiltration density, especially in those with EBV-associated gastric cancer, which may direct patient selection for checkpoint blockade therapy.

  3. A nonimmunogenic sarcoma transduced with the cDNA for interferon gamma elicits CD8+ T cells against the wild-type tumor: correlation with antigen presentation capability

    PubMed Central

    1992-01-01

    To be recognized by CD8+ T lymphocytes, target cells must process and present peptide antigens in the context of major histocompatibility complex (MHC) class I molecules. The nonimmunogenic, low class I- expressing, methylcholanthrene (MCA)-induced murine sarcoma cell line, MCA 101, is a poor presenter of endogenously generated viral antigens to specific CD8+ T lymphocytes and cannot be used to generate tumor infiltrating lymphocytes (TIL). Since interferon gamma (IFN-gamma) has been shown to upregulate three sets of molecules important for antigen processing and presentation, we retrovirally transduced wild-type MCA 101 (101.WT) tumor with the mIFN-gamma cDNA to create the 101.NAT cell line. Unlike 101.WT, some clones of retrovirally transduced 101.NAT tumor expressed high levels of class I, and could be used to generate CD8+ TIL. More importantly, these TIL were therapeutic in vivo against established pulmonary metastases from the wild-type tumor. Although not uniformly cytotoxic amongst several separate cultures, these TIL did specifically release cytokines (IFN-gamma and tumor necrosis factor- alpha) in response to 101.WT targets. 101.WT's antigen presentation deficit was also reversed by gene modification with mIFN-gamma cDNA. 101.NAT had a greatly improved capacity to present viral antigens to CD8+ cytotoxic T lymphocytes. These findings show that a nonimmunogenic tumor, incapable of generating a CD8+ T cell immune response, could be gene-modified to generate a therapeutically useful immune response against the wild-type tumor. This strategy may be useful in developing treatments for tumor histologies not thought to be susceptible to T cell-based immunotherapy. PMID:1588273

  4. PD1 and PDL1 expression in primary central nervous system diffuse large B-cell lymphoma are frequent and expression of PD1 predicts poor survival.

    PubMed

    Four, Marion; Cacheux, Valère; Tempier, Ariane; Platero, Dolorès; Fabbro, Michel; Marin, Grégory; Leventoux, Nicolas; Rigau, Valérie; Costes-Martineau, Valérie; Szablewski, Vanessa

    2016-12-13

    Primary central nervous system diffuse large B-cell lymphoma (PCNS-DLBCL) is a rare and aggressive type of diffuse large B-cell lymphoma (DLBCL) whit poorly understood pathogenesis. Finding biomarkers associated with patient survival may be important for understanding its physiopathology and to develop new therapeutic approaches. We investigated 32 PCNS-DLBCL from immunocompetent patients for BCL2, CMYC, LMO2, and P53 expression and for cytogenetic aberrations of BCL2, BCL6, and MYC genes, all known for their prognostic value in systemic DLBCL (s-DLBCL). We analyzed PD1 and PDL1 protein expression in both tumor infiltrating lymphocytes (TILs) and tumor cells. Finally, we searched for correlation between biological data and clinical course. The PCNS-DLBCL expressed BCL2, CMYC, LMO2, and P53 at similar frequency than s-DLBCL but without significant prognostic on survival. None cases harbored aberrations involving BCL2 and MYC gene whereas BCL6 abnormalities were present in 20.7% of cases but without value on survival. Expression of PD1 in TILs and PDL1 in tumor cells was observed at higher rates than in s-DLBCL (58% and 37%, respectively). The PD1 expression in TILs correlated with PDL1 expression in tumor cells (P = .001). Presence of PD1 positive TILs was associated with poorer overall survival (P = .011). Patients with PDL1 overexpression tended to better response to chemotherapy (P = .23). In conclusion PCNS-DLBCL pathogenesis differs from s-DLBCL without prognostic value of the phenotypic and cytogenetic parameters known for their pejorative impact in the latter. The PD1/PDL1 pathway plays a strong role in PCNS-DLBCL and represents an attractive target for this aggressive lymphoma.

  5. Programmed cell death ligand 1 expression in osteosarcoma.

    PubMed

    Shen, Jacson K; Cote, Gregory M; Choy, Edwin; Yang, Pei; Harmon, David; Schwab, Joseph; Nielsen, G Petur; Chebib, Ivan; Ferrone, Soldano; Wang, Xinhui; Wang, Yangyang; Mankin, Henry; Hornicek, Francis J; Duan, Zhenfeng

    2014-07-01

    Programmed cell death ligand 1 (PDL1, also known as B7H1) is a cell-surface protein that suppresses the cytotoxic CD8(+) T-cell-mediated immune response. PDL1 expression and its clinical relevance in sarcomas are not well understood. Therefore, we sought to measure RNA expression levels for PDL1 in 38 clinically annotated osteosarcoma tumor samples and aimed to determine if PDL1 expression correlates with clinical features and tumor-infiltrating lymphocytes (TIL). Quantitative real-time RT-PCR for PDL1 was optimized in 18 cell lines, of which 5 were osteosarcoma derived. qRT-PCR results were validated via flow cytometry and immunohistochemistry (IHC) in select cell lines. Total RNA was isolated from 38 human osteosarcoma samples for qRT-PCR analysis. Clinical data were sorted, and significance was determined by the Student t test. TILs were examined in patient samples by tissue microarray hematoxylin-eosin staining. We confirmed the constitutive PDL1 mRNA expression in cell lines by qRT-PCR, flow cytometry, and IHC. Across human osteosarcoma samples, PDL1 mRNA gene expression ranged over 4 log (>5,000-fold difference). Relative expression levels were evaluated against clinical factors such as age/gender, metastasis, recurrence, chemotherapy, percentage of necrosis, and survival; no significant associations were identified. The presence of TILs was associated with high PDL1 expression (R(2) = 0.37; P = 0.01). In summary, we developed an RNA-based assay to determine PDL1 expression levels, and we show, for the first time, that high levels of PDL1 are expressed in a subset of osteosarcoma, and PDL1 expression is positively correlated with TILs. Multiple agents targeting PD1/PDL1 are in clinical development, and this may be a novel immunotherapeutic strategy for osteosarcoma clinical trials.

  6. Expression of inhibitory receptors on intratumoral T cells modulates the activity of a T cell-bispecific antibody targeting folate receptor.

    PubMed

    Schreiner, Jens; Thommen, Daniela S; Herzig, Petra; Bacac, Marina; Klein, Christian; Roller, Andreas; Belousov, Anton; Levitsky, Victor; Savic, Spasenija; Moersig, Wolfgang; Uhlenbrock, Franziska; Heinzelmann-Schwarz, Viola A; Umana, Pablo; Pisa, Pavel; von Bergwelt-Baildon, M; Lardinois, Didier; Müller, Philipp; Karanikas, Vaios; Zippelius, Alfred

    2016-02-01

    T-cell bispecific antibodies (TCBs) are a novel therapeutic tool designed to selectively recruit T-cells to tumor cells and simultaneously activate them. However, it is currently unknown whether the dysfunctional state of T-cells, embedded into the tumor microenvironment, imprints on the therapeutic activity of TCBs. We performed a comprehensive analysis of activation and effector functions of tumor-infiltrating T-cells (TILs) in different tumor types, upon stimulation by a TCB targeting folate receptor 1 and CD3 (FolR1-TCB). We observed a considerable heterogeneity in T-cell activation, cytokine production and tumor cell killing upon exposure to FolR1-TCB among different FolR1-expressing tumors. Of note, tumors presenting with a high frequency of PD-1(hi) TILs displayed significantly impaired tumor cell killing and T-cell function. Further characterization of additional T-cell inhibitory receptors revealed that PD-1(hi) TILs defined a T-cell subset with particularly high levels of multiple inhibitory receptors compared with PD-1(int) and PD-1(neg) T-cells. PD-1 blockade could restore cytokine secretion but not cytotoxicity of TILs in a subset of patients with scarce PD-1(hi) expressing cells; in contrast, patients with abundance of PD-1(hi) expressing T-cells did not benefit from PD-1 blockade. Our data highlight that FolR1-TCB is a promising novel immunotherapeutic treatment option which is capable of activating intratumoral T-cells in different carcinomas. However, its therapeutic efficacy may be substantially hampered by a pre-existing dysfunctional state of T-cells, reflected by abundance of intratumoral PD-1(hi) T-cells. These findings present a rationale for combinatorial approaches of TCBs with other therapeutic strategies targeting T-cell dysfunction.

  7. Sustainable Contingency Base Camp Operations

    DTIC Science & Technology

    2010-06-17

    L , IJU;TAL 6:1 6>::’JI&O.’ID’ C unums - IVA’TU.Al.V.$ - -~ D Tli,Oil!’tJ Mi’(’(n ~p t::il A•ilt’UJI -~· G_jJ "’"n - AJO ?Solti’U.E - AtT""llq

  8. Physiochemical/Rheological Control of Nonmetallic Materials.

    DTIC Science & Technology

    1982-08-01

    Temperature Tge Temperature at Gel T Glass Transition Temperature rl Viscosity t gel Time to Gel G’ Storage Modulus G" Loss Modulus til Enthalpy...standards were obtained and the amounts of the volatiles were determined by gas chromotography . Where applicable, the thermal and rheological properties...aluminum powder and the fillers. The filtrate , which contained th two epoxies and DICY, cured without any foaming. Attempts to separate the exact blowing

  9. Lymphocyte imprinting with melanoma antigens acquired by trogocytosis facilitates identification of tumor-reactive T cells

    PubMed Central

    Eisenberg, Galit; Uzana, Ronny; Pato, Aviad; Frankenburg, Shoshana; Merims, Sharon; Yefenof, Eitan; Ferrone, Soldano; Peretz, Tamar; Machlenkin, Arthur; Lotem, Michal

    2013-01-01

    Trogocytosis is a contact-dependent inter-cellular transfer of membrane fragments and associated molecules from antigen presenting cells to effector lymphocytes. We previously demonstrated that trogocytosis also occurs between tumor target and cognate melanoma antigen-specific cytotoxic T cells (CTL). Here we show that, following trogocytosis, immune effector cells acquire molecular components of the tumor, including surface antigens, which are detectable by specific monoclonal antibodies. We demonstrate that CD8+ and CD4+ T cells from melanoma patients’ PBMC and tumor infiltrating lymphocytes (TIL) capture melanoma antigens, enabling identification of trogocytosing lymphocytes by staining with antigen-specific antibodies. This finding circumvents the necessity of tumor pre-labeling, which in the past was mandatory to detect membrane-capturing T cells. Through the detection of melanoma antigens on TIL, we sorted trogocytosing T cells and verified their preferential reactivity and cytotoxicity. Furthermore, tumor-antigen imprinted T cells were detected at low frequency in fresh TIL cultures shortly after extraction from the tumor. Thus, T cell imprinting by tumor antigens may allow the enrichment of melanoma antigen-specific T cells for research and potentially even for the adoptive immunotherapy of patients with cancer. PMID:23626012

  10. PD-L1 expression on immune cells, but not on tumor cells, is a favorable prognostic factor for head and neck cancer patients

    PubMed Central

    Kim, Hye Ryun; Ha, Sang-Jun; Hong, Min Hee; Heo, Su Jin; Koh, Yoon Woo; Choi, Eun Chang; Kim, Eun Kyung; Pyo, Kyoung Ho; Jung, Inkyung; Seo, Daekwan; Choi, Jaewoo; Cho, Byoung Chul; Yoon, Sun Och

    2016-01-01

    To investigate the expression of programmed death-ligand 1 (PD-L1) and immune checkpoints and their prognostic value for resected head and neck squamous cell cancer (HNSCC). PD-L1 expression on tumor cells (TC) and tumor-infiltrating immune cells (IC), abundance of tumor-infiltrating lymphocytes (TILs), and expression of the immune checkpoints were investigated in 402 HNSCC patients. PD-L1 expression on TC and IC was categorized into four groups according to the percentage of PD-L1-positive cells. PD-L1 positivity was defined as ≥5% of cells based on immunohistochemistry. High PD-L1 expression on IC, but not TC, was an independent favorable prognostic factor for RFS and OS adjusted for age, gender, smoking, stage, and HPV. High frequencies of CD3+ or CD8+ TILs, Foxp3+ Tregs, and PD-1+ TILs were strongly associated with favorable prognosis. PD-L1 was exclusively expressed on either TC or IC. Transcriptome analysis demonstrated that IC3 expressed higher levels of the effector T cell markers than TC3, suggesting that PD-L1 expression is regulated via an adaptive IFNγ-mediated mechanism. High PD-L1 expression on IC, but not TC, and high abundance of PD-1+ T cells and Foxp3+ Tregs are favorable prognostic factors for resected HNSCC. This study highlights the importance of comprehensive assessment of both TC and IC. PMID:27841362

  11. Diversity and divergence of the glioma-infiltrating T-cell receptor repertoire

    PubMed Central

    Sims, Jennifer S.; Grinshpun, Boris; Feng, Yaping; Ung, Timothy H.; Neira, Justin A.; Samanamud, Jorge L.; Canoll, Peter; Shen, Yufeng; Sims, Peter A.; Bruce, Jeffrey N.

    2016-01-01

    Although immune signaling has emerged as a defining feature of the glioma microenvironment, how the underlying structure of the glioma-infiltrating T-cell population differs from that of the blood from which it originates has been difficult to measure directly in patients. High-throughput sequencing of T-cell receptor (TCR) repertoires (TCRseq) provides a population-wide statistical description of how T cells respond to disease. We have defined immunophenotypes of whole repertoires based on TCRseq of the α- and β-chains from glioma tissue, nonneoplastic brain tissue, and peripheral blood from patients. Using information theory, we partitioned the diversity of these TCR repertoires into that from the distribution of VJ cassette combinations and diversity due to VJ-independent factors, such as selection due to antigen binding. Tumor-infiltrating lymphocytes (TILs) possessed higher VJ-independent diversity than nonneoplastic tissue, stratifying patients according to tumor grade. We found that the VJ-independent components of tumor-associated repertoires diverge more from their corresponding peripheral repertoires than T-cell populations in nonneoplastic brain tissue, particularly for low-grade gliomas. Finally, we identified a “signature” set of TCRs whose use in peripheral blood is associated with patients exhibiting low TIL divergence and is depleted in patients with highly divergent TIL repertoires. This signature is detectable in peripheral blood, and therefore accessible noninvasively. We anticipate that these immunophenotypes will be foundational to monitoring and predicting response to antiglioma vaccines and immunotherapy. PMID:27261081

  12. Feature-based registration of historical aerial images by Area Minimization

    NASA Astrophysics Data System (ADS)

    Nagarajan, Sudhagar; Schenk, Toni

    2016-06-01

    The registration of historical images plays a significant role in assessing changes in land topography over time. By comparing historical aerial images with recent data, geometric changes that have taken place over the years can be quantified. However, the lack of ground control information and precise camera parameters has limited scientists' ability to reliably incorporate historical images into change detection studies. Other limitations include the methods of determining identical points between recent and historical images, which has proven to be a cumbersome task due to continuous land cover changes. Our research demonstrates a method of registering historical images using Time Invariant Line (TIL) features. TIL features are different representations of the same line features in multi-temporal data without explicit point-to-point or straight line-to-straight line correspondence. We successfully determined the exterior orientation of historical images by minimizing the area formed between corresponding TIL features in recent and historical images. We then tested the feasibility of the approach with synthetic and real data and analyzed the results. Based on our analysis, this method shows promise for long-term 3D change detection studies.

  13. Very Late Antigen-1 Marks Functional Tumor-Resident CD8 T Cells and Correlates with Survival of Melanoma Patients

    PubMed Central

    Murray, Timothy; Fuertes Marraco, Silvia A.; Baumgaertner, Petra; Bordry, Natacha; Cagnon, Laurène; Donda, Alena; Romero, Pedro; Verdeil, Grégory; Speiser, Daniel E.

    2016-01-01

    A major limiting factor in the success of immunotherapy is tumor infiltration by CD8+ T cells, a process that remains poorly understood. In the present study, we characterized homing receptors expressed by human melanoma-specific CD8+ T cells. Our data reveal that P-selectin binding and expression of the retention integrin, very late antigen (VLA)-1, by vaccine-induced T cells correlate with longer patient survival. Furthermore, we demonstrate that CD8+VLA-1+ tumor-infiltrating lymphocytes (TILs) are highly enriched in melanoma metastases in diverse tissues. VLA-1-expressing TIL frequently co-express CD69 and CD103, indicating tissue-resident memory T cells (TRM) differentiation. We employed a mouse model of melanoma to further characterize VLA-1-expressing TIL. Our data show that VLA-1+ TRM develop in murine tumors within 2 weeks, where they exhibit increased activation status, as well as superior effector functions. In addition, in vivo blockade of either VLA-1 or CD103 significantly impaired control of subcutaneous tumors. Together, our data indicate that VLA-1+ TRM develop in tumors and play an important role in tumor immunity, presenting novel targets for the optimization of cancer immunotherapy. PMID:28018343

  14. Transport Mechanism of Nicotine in Primary Cultured Alveolar Epithelial Cells.

    PubMed

    Takano, Mikihisa; Nagahiro, Machi; Yumoto, Ryoko

    2016-02-01

    Nicotine is absorbed from the lungs into the systemic circulation during cigarette smoking. However, there is little information concerning the transport mechanism of nicotine in alveolar epithelial cells. In this study, we characterized the uptake of nicotine in rat primary cultured type II (TII) and transdifferentiated type I-like (TIL) epithelial cells. In both TIL and TII cells, [(3)H]nicotine uptake was time and temperature-dependent, and showed saturation kinetics. [(3)H]Nicotine uptake in these cells was not affected by Na(+), but was sensitive to extracellular and intracellular pH, suggesting the involvement of a nicotine/proton antiport system. The uptake of [(3)H]nicotine in these cells was potently inhibited by organic cations such as clonidine, diphenhydramine, and pyrilamine, but was not affected by substrates and/or inhibitors of known organic cation transporters such as carnitine, 1-methyl-4-phenylpyridinium, and tetraethylammonium. In addition, the uptake of [(3)H]nicotine in TIL cells was stimulated by preloading the cells with unlabeled nicotine, pyrilamine, and diphenhydramine, but not with tetraethylammonium. These results suggest that a novel proton-coupled antiporter is involved in the uptake of nicotine in alveolar epithelial cells and its absorption from the lungs into the systemic circulation.

  15. High incidence of interleukin 10 mRNA but not interleukin 2 mRNA detected in human breast tumours.

    PubMed Central

    Venetsanakos, E.; Beckman, I.; Bradley, J.; Skinner, J. M.

    1997-01-01

    Despite the presence of a lymphocytic infiltrate in solid cancers, the failure for tumour growth to be contained suggests an inadequate immune response to the tumour. Poor cytotoxicity exerted by tumour-infiltrating lymphocytes (TILs) against tumour cells in vitro, combined with continued tumour growth in vivo, suggests deficiencies in TIL function or numbers. Various theories have been postulated to explain how tumour cells may escape immunosurveillance and control. One of the many hypotheses is the failure of production of cytokines, which are necessary for T cells to mediate their function. Thus, the expression of cytokine mRNA in human breast tumour sections was investigated by reverse transcriptase polymerase chain reaction (RT-PCR) with cytokine-specific primers. A relatively consistent finding was detection of interleukin (IL) 10 mRNA among the tumours. No IL-2 and little IL-4 mRNA was detected in the tumours. IL-6 and IL-10 mRNA was detected in only one and two of the normal breast tissues respectively. IL-2, IL-4 and tumour necrosis factor (TNF)-alpha mRNA was not detected in any of the normal breast tissues. The reduced function of TILs may be related to IL-10, which has known inhibitory effects on T-cell activation. Images Figure 1 PMID:9192989

  16. Kupffer cells of cirrhotic rat livers sensitize colon cancer cells to Fas-mediated apoptosis.

    PubMed

    Song, E; Chen, J; Ouyang, N; Wang, M; Exton, M S; Heemann, U

    2001-05-04

    Metastasis of colorectal carcinomas rarely occurs in cirrhotic livers. Our study investigated the influence of activated Kupffer cells from cirrhotic rat livers on hepatic colonization and FasR-mediated apoptosis of colon cancer cells. A rat colon cancer cell line, RCN-9, was used to inoculate rat livers. Treatment with conditioned media of Kupffer cells isolated from CCl(4)-induced cirrhotic rat livers (cirrhotic KCM) significantly reduced the incidence of hepatic colonization of RCN-9 cells. In vitro cytotoxicity of Kupffer cells and tumour infiltrating lymphocytes (TILs) on RCN-9 cells was evaluated using [(3)H]-release assay. RCN-9 cells were resistant to cytotoxicity mediated by cirrhotic Kupffer cells, but were sensitized to TIL-mediated killing after treatment with cirrhotic KCM. The specific killing induced by TILs was FasR-mediated, as it was inhibited by ZB4, an antagonistic anti-FasR antibody. In agreement, cirrhotic KCM increased recombinant Fas ligand-induced apoptosis of RCN-9 cells, and up-regulated FasR expression on RCN-9 cells as evaluated by RT-PCR and flow cytometry. These findings suggest that Kupffer cells in cirrhotic livers sensitize metastatic colon cancer cells to FasR-mediated apoptosis by up-regulating the receptors, which thus prepare them to be eliminated by infiltrating lymphocytes.

  17. Tumor infiltrating lymphocyte therapy for ovarian cancer and renal cell carcinoma

    PubMed Central

    Andersen, Rikke; Donia, Marco; Westergaard, Marie Christine Wulff; Pedersen, Magnus; Hansen, Morten; Svane, Inge Marie

    2015-01-01

    Personalized cancer immunotherapy based on infusion of T cells holds the promise to specifically target a patient’s individual tumor. Accumulating evidence indicates that the T cells mediating these tumor regressions after cancer immunotherapies may primarily target patient-specific mutations expressed by the patients’ tumors and that the presence of these “neo-antigen” specific T-cells may be related to a high number of mutations in the tumor. In melanoma, treatment with autologous tumor-infiltrating lymphocytes (TILs) can mediate durable complete responses. Previous trials investigating TIL therapy in solid tumors other than melanoma have shown limited success, however none of these early trials used current preparative chemotherapy regimens, and the methods for in vitro lymphocyte expansion have changed considerably. New advances and understandings in T cell based immunotherapies have stimulated the interest in developing this approach for other indications. Here, we summarize the early clinical data in the field of adoptive cell transfer therapy (ACT) using tumor-infiltrating lymphocytes for patients with renal cell carcinoma (RCC) and ovarian cancer (OC). In addition we describe the major advances in the characterization and application of TIL therapy for patients with RCC and OC. PMID:26308285

  18. Lymphocyte-to-monocyte ratio is associated with prognosis of diffuse large B-cell lymphoma: correlation with CD163 positive M2 type tumor-associated macrophages, not PD-1 positive tumor-infiltrating lymphocytes.

    PubMed

    Wang, Jingxuan; Gao, Kun; Lei, Wanting; Dong, Lina; Xuan, Qijia; Feng, Meiyan; Wang, Jinlu; Ye, Xiangnan; Jin, Tuan; Zhang, Zhongbai; Zhang, Qingyuan

    2017-01-17

    The research aims to examine the prognostic value of the lymphocyte-to-monocyte ratio (LMR), neutrophil-to- lymphocyte ratio (NLR) and platelet-to-lymphocyte ratio (PLR) in diffuse large B-cell lymphoma (DLBCL). The relation of these hematologic indicators to poor antitumor immunity and prognosis must be investigated. Clinicopathologic data and survival information of 355 patients with DLBCL was retrospectively analyzed. Univariate analysis revealed that lower LMR (<2.71), higher NLR (≥2.81), CD163+ M2 tumor-associated macrophages (TAM) content ≥9.5% and programmed cell death 1 (PD-1)+ tumor-infiltrating lymphocytes (TILs) content < 4.5 cells per high power field(HPF) were significantly related to unfavorable overall survival (OS) and progression free survival (PFS). When considering the prognostic indexes of IPI, multivariate analysis confirmed that LMR of <2.71 and CD163+ M2 TAM content ≥9.5% significantly affected the prognosis of DLBCL. Spearman correlation test showed LMR was negatively correlated with CD163+ M2 TAM content. However, there were no correlation was found between LMR and PD-1+ TIL as well as between NLR and PD-1+ TIL content. These results indicated that decreased LMR lead to a weak anti-tumor immunity and could be used as a bad prognosis biomarker of DLBCL.

  19. Quantification of image quality using information theory.

    PubMed

    Niimi, Takanaga; Maeda, Hisatoshi; Ikeda, Mitsuru; Imai, Kuniharu

    2011-12-01

    Aims of present study were to examine usefulness of information theory in visual assessment of image quality. We applied first order approximation of the Shannon's information theory to compute information losses (IL). Images of a contrast-detail mammography (CDMAM) phantom were acquired with computed radiographies for various radiation doses. Information content was defined as the entropy Σp( i )log(1/p ( i )), in which detection probabilities p ( i ) were calculated from distribution of detection rate of the CDMAM. IL was defined as the difference between information content and information obtained. IL decreased with increases in the disk diameters (P < 0.0001, ANOVA) and in the radiation doses (P < 0.002, F-test). Sums of IL, which we call total information losses (TIL), were closely correlated with the image quality figures (r = 0.985). TIL was dependent on the distribution of image reading ability of each examinee, even when average reading ratio was the same in the group. TIL was shown to be sensitive to the observers' distribution of image readings and was expected to improve the evaluation of image quality.

  20. Immune-mediated antitumor effect by type 2 diabetes drug, metformin

    PubMed Central

    Eikawa, Shingo; Nishida, Mikako; Mizukami, Shusaku; Yamazaki, Chihiro; Nakayama, Eiichi; Udono, Heiichiro

    2015-01-01

    Metformin, a prescribed drug for type 2 diabetes, has been reported to have anti-cancer effects; however, the underlying mechanism is poorly understood. Here we show that this mechanism may be immune-mediated. Metformin enabled normal but not T-cell–deficient SCID mice to reject solid tumors. In addition, it increased the number of CD8+ tumor-infiltrating lymphocytes (TILs) and protected them from apoptosis and exhaustion characterized by decreased production of IL-2, TNFα, and IFNγ. CD8+ TILs capable of producing multiple cytokines were mainly PD-1−Tim-3+, an effector memory subset responsible for tumor rejection. Combined use of metformin and cancer vaccine improved CD8+ TIL multifunctionality. The adoptive transfer of antigen-specific CD8+ T cells treated with metformin concentrations as low as 10 μM showed efficient migration into tumors while maintaining multifunctionality in a manner sensitive to the AMP-activated protein kinase (AMPK) inhibitor compound C. Therefore, a direct effect of metformin on CD8+ T cells is critical for protection against the inevitable functional exhaustion in the tumor microenvironment. PMID:25624476

  1. Tumor-infiltrating lymphocytes are dynamically desensitized to antigen but are maintained by homeostatic cytokine

    PubMed Central

    Boldajipour, Bijan; Nelson, Amanda; Krummel, Matthew F.

    2016-01-01

    T cells that enter tumors are largely tolerized, but how that process is choreographed and how the ensuing “dysfunctional” tumor-infiltrating lymphocytes (TILs) are maintained are poorly understood and are difficult to assess in spontaneous disease. We exploited an autochthonous model of breast cancer for high-resolution imaging of the early and later stages of tumor residence to understand the relationships between cellular behaviors and cellular phenotypes. “Dysfunctional” differentiation began within the first days of tumor residence with an initial phase in which T cells arrest, largely on tumor-associated macrophages. Within 10 days, cellular motility increased and resembled a random walk, suggesting a relative absence of TCR signaling. We then studied the concurrent and apparently contradictory phenomenon that many of these cells express molecular markers of activation and were visualized undergoing active cell division. We found that whereas proliferation did not require ongoing TCR/ZAP70 signaling, instead this is driven in part by intratumoral IL-15 cytokine. Thus, TILs undergo sequential reprogramming by the tumor microenvironment and are actively retained, even while being antigen insensitive. We conclude that this program effectively fills the niche with ineffective yet cytokine-dependent TILs, and we propose that these might compete with new clones, when they arise. PMID:27942588

  2. CD74 and intratumoral immune response in breast cancer.

    PubMed

    Wang, Zhi-Qiang; Milne, Katy; Webb, John R; Watson, Peter H

    2016-04-06

    CD74 (invariant chain) plays a role in MHC class II antigen presentation. We assessed CD74 and MHCII expression in tumor cells, as well as CD8, CD4, and CD68 tumor infiltrating leucocyte (TIL) density by immunohistochemistry in a cohort of 492 breast cancer patients. CD74 expression was associated with poor prognostic markers including patient age, tumor grade, ER status, non-Luminal A subtypes, and with MHCII expression and higher TIL densities, particularly in the Basal-like subgroup. Univariate analysis showed a favorable prognostic effect of CD74 (Hazard ratio = 0.46, 95% CI = 0.26-0.89, p = 0.022) and for combined CD74/MHCII (Hazard ratio = 0.26, 95% CI = 0.17-0.81, p = 0.014) positive status for overall survival that was only manifested in the Basal-like subgroup. CD74 and MHCII expression is associated with patient survival in Basal-like breast cancer, and the association with TIL may reflect an effective intratumoral immune response.

  3. Identification of genes and pathways associated with cytotoxic T lymphocyte infiltration of serous ovarian cancer

    PubMed Central

    Leffers, N; Fehrmann, R S N; Gooden, M J M; Schulze, U R J; ten Hoor, K A; Hollema, H; Boezen, H M; Daemen, T; de Jong, S; Nijman, H W; van der Zee, A G J

    2010-01-01

    Background: Tumour-infiltrating lymphocytes (TILs) are predictors of disease-specific survival (DSS) in ovarian cancer. It is largely unknown what factors contribute to lymphocyte recruitment. Our aim was to evaluate genes and pathways contributing to infiltration of cytotoxic T lymphocytes (CTLs) in advanced-stage serous ovarian cancer. Methods: For this study global gene expression was compared between low TIL (n=25) and high TIL tumours (n=24). The differences in gene expression were evaluated using parametric T-testing. Selectively enriched biological pathways were identified with gene set enrichment analysis. Prognostic influence was validated in 157 late-stage serous ovarian cancer patients. Using immunohistochemistry, association of selected genes from identified pathways with CTL was validated. Results: The presence of CTL was associated with 320 genes and 23 pathways (P<0.05). In addition, 54 genes and 8 pathways were also associated with DSS in our validation cohort. Immunohistochemical evaluation showed strong correlations between MHC class I and II membrane expression, parts of the antigen processing and presentation pathway, and CTL recruitment. Conclusion: Gene expression profiling and pathway analyses are valuable tools to obtain more understanding of tumour characteristics influencing lymphocyte recruitment in advanced-stage serous ovarian cancer. Identified genes and pathways need to be further investigated for suitability as therapeutic targets. PMID:20664601

  4. Effects of Short-Term Thermal Alteration on Organic Matter in Experimentally-Heated Tagish Lake Observed by Raman Spectroscopy

    NASA Technical Reports Server (NTRS)

    Chan, Q. H. S.; Nakato, A.; Zolensky, M. E.; Nakamura, T.; Kebukawa, Y.

    2007-01-01

    Carbonaceous chondrites exhibit a wide range of aqueous and thermal alteration characteristics. Examples of the thermally metamorphosed carbonaceous chondrites (TMCCs) include the C2-ung/CM2TIVs Belgica (B)-7904 and Yamato (Y) 86720. The alteration extent is the most complete in these meteorites and thus they are considered typical end-members of TMCCs exhibiting complete dehydration of matrix phyllosilicates [1, 2]. The estimated heating conditions are 10 to 10(sup 3) days at 700 C to 1 to 100 hours at 890 C, i.e. short-term heating induced by impact and/or solar radiation [3]. The chemical and bulk oxygen isotopic compositions of the matrix of the carbonate (CO3)-poor lithology of the Tagish Lake (hereafter Tag) meteorite bears similarities to these TMCCs [4]. We investigated the experimentally-heated Tag with the use of Raman spectroscopy to understand how short-term heating affects the maturity of insoluble organic matter (IOM) in aqueously altered meteorites.

  5. HIV-1 Vpr degrades the HLTF DNA translocase in T cells and macrophages

    PubMed Central

    Lahouassa, Hichem; Blondot, Marie-Lise; Chauveau, Lise; Chougui, Ghina; Morel, Marina; Leduc, Marjorie; Guillonneau, François; Ramirez, Bertha Cecilia; Schwartz, Olivier; Margottin-Goguet, Florence

    2016-01-01

    Viruses often interfere with the DNA damage response to better replicate in their hosts. The human immunodeficiency virus 1 (HIV-1) viral protein R (Vpr) protein has been reported to modulate the activity of the DNA repair structure-specific endonuclease subunit (SLX4) complex and to promote cell cycle arrest. Vpr also interferes with the base-excision repair pathway by antagonizing the uracil DNA glycosylase (Ung2) enzyme. Using an unbiased quantitative proteomic screen, we report that Vpr down-regulates helicase-like transcription factor (HLTF), a DNA translocase involved in the repair of damaged replication forks. Vpr subverts the DDB1–cullin4-associated-factor 1 (DCAF1) adaptor of the Cul4A ubiquitin ligase to trigger proteasomal degradation of HLTF. This event takes place rapidly after Vpr delivery to cells, before and independently of Vpr-mediated G2 arrest. HLTF is degraded in lymphocytic cells and macrophages infected with Vpr-expressing HIV-1. Our results reveal a previously unidentified strategy for HIV-1 to antagonize DNA repair in host cells. PMID:27114546

  6. Strand-biased cytosine deamination at the replication fork causes cytosine to thymine mutations in Escherichia coli.

    PubMed

    Bhagwat, Ashok S; Hao, Weilong; Townes, Jesse P; Lee, Heewook; Tang, Haixu; Foster, Patricia L

    2016-02-23

    The rate of cytosine deamination is much higher in single-stranded DNA (ssDNA) than in double-stranded DNA, and copying the resulting uracils causes C to T mutations. To study this phenomenon, the catalytic domain of APOBEC3G (A3G-CTD), an ssDNA-specific cytosine deaminase, was expressed in an Escherichia coli strain defective in uracil repair (ung mutant), and the mutations that accumulated over thousands of generations were determined by whole-genome sequencing. C:G to T:A transitions dominated, with significantly more cytosines mutated to thymine in the lagging-strand template (LGST) than in the leading-strand template (LDST). This strand bias was present in both repair-defective and repair-proficient cells and was strongest and highly significant in cells expressing A3G-CTD. These results show that the LGST is accessible to cellular cytosine deaminating agents, explains the well-known GC skew in microbial genomes, and suggests the APOBEC3 family of mutators may target the LGST in the human genome.

  7. A multiplex assay based on encoded microbeads conjugated to DNA NanoBeacons to monitor base excision repair activities by flow cytometry.

    PubMed

    Gines, Guillaume; Saint-Pierre, Christine; Gasparutto, Didier

    2014-08-15

    We reported here a new assay to detect base excision repair activities from purified enzymes, as well as in cell-free extracts. The multiplex format rests upon the encoding of magnetic beads with the fluorophore Alexa 488, thanks to a fast and original procedure. Fluorescently encoded microbeads are subsequently functionalized by lesion-containing DNA NanoBeacons labeled with the fluorophore/quencher pair Cyanine 5/BHQ2. Probes cleavage, induced by targeted enzymes leads to Cyanine 5 signal enhancement, which is finally quantified by flow cytometry. The multiplex assay was applied to the detection of restriction enzymes activities as well as base excision repair processes. Each test requires only 500fmol of DNA substrate, which constitutes great sensitivity compared to other BER functional assays. The present biosensor is able to detect both uracil DNA N-glycosylase (UNG) and AP-endonuclease 1 (APE1) within few nanograms of nuclear extract. Additionally, we demonstrated that the corresponding assay has potential application in DNA repair inhibitor search. Finally, the current multiplexed tool shows several advantages in comparison to other functional BER assays with no need of electrophoretic separation, straightforward, easy and reproducible functionalization of encoded microbeads and a high stability of DNA probes in cell-free extracts.

  8. Modellgetriebene Softwareentwicklung für Robotiksysteme

    NASA Astrophysics Data System (ADS)

    Steck, Andreas; Stampfer, Dennis; Schlegel, Christian

    Die Erschließung breiter Anwendungspotenziale für Serviceroboter erfordert den Schritt weg von manuell erstellten Einzelentwürfen hin zu baukastenartig zusammengesetzten Systemen. Grundlegend hierfür ist der Schritt weg von codezentrierten hin zu modellgetriebenen Systemen. In modellgetriebenen Ansätzen wird langlebiges Lösungswissen von kurzlebigen Implementierungstechnologien entkoppelt. Durch das Explizieren von Eigenschaften wie Ressourcenbedarf und Kommunikationsverhalten wird die Systemebene adressiert, so dass Zusammensetzbarkeit von ausgereiften Komponenten ebenso unterstützt werden kann wie der Nachweis von Systemeigenschaften. Die so möglichen ressourcenadäquaten Lösungen mit zugesicherten Eigenschaften und der Wiederverwendung ausgereifter Lösungen wird als grundlegend für die effiziente Umsetzung der geforderten Qualitätsmaßstäbe vieler zukünftiger Servicerobotikapplikationen gesehen. Diese Arbeit beschreibt die Umsetzung eines modellgetriebenen Softwareentwicklungsprozesses in OpenArchitectureWare auf der Basis eines Komponentenansatzes und eines Ausführungscontainers, der beispielsweise völlig unabhängig von der Middleware ist.

  9. Application of the Firefly and Luus-Jaakola algorithms in the calculation of a double reactive azeotrope

    NASA Astrophysics Data System (ADS)

    Mendes Platt, Gustavo; Pinheiro Domingos, Roberto; Oliveira de Andrade, Matheus

    2014-01-01

    The calculation of reactive azeotropes is an important task in the preliminary design and simulation of reactive distillation columns. Classically, homogeneous nonreactive azeotropes are vapor-liquid coexistence conditions where phase compositions are equal. For homogeneous reactive azeotropes, simultaneous phase and chemical equilibria occur concomitantly with equality of compositions (in the Ung-Doherty transformed space). The modeling of reactive azeotrope calculation is represented by a nonlinear algebraic system with phase equilibrium, chemical equilibrium and azeotropy equations. This nonlinear system can exhibit more than one solution, corresponding to a double reactive azeotrope. In a previous paper (Platt et al 2013 J. Phys.: Conf. Ser. 410 012020), we investigated some numerical aspects of the calculation of reactive azeotropes in the isobutene + methanol + methyl-tert-butyl-ether (with two reactive azeotropes) system using two metaheuristics: the Luus-Jaakola adaptive random search and the Firefly algorithm. Here, we use a hybrid structure (stochastic + deterministic) in order to produce accurate results for both azeotropes. After identifying the neighborhood of the reactive azeotrope, the nonlinear algebraic system is solved using Newton's method. The results indicate that using metaheuristics and some techniques devoted to the calculation of multiple minima allows both azeotropic coordinates in this reactive system to be obtains. In this sense, we provide a comprehensive analysis of a useful framework devoted to solving nonlinear systems, particularly in phase equilibrium problems.

  10. DNA Damage Responses Are Induced by tRNA Anticodon Nucleases and Hygromycin B.

    PubMed

    Wemhoff, Sabrina; Klassen, Roland; Beetz, Anja; Meinhardt, Friedhelm

    2016-01-01

    Previous studies revealed DNA damage to occur during the toxic action of PaT, a fungal anticodon ribonuclease (ACNase) targeting the translation machinery via tRNA cleavage. Here, we demonstrate that other translational stressors induce DNA damage-like responses in yeast as well: not only zymocin, another ACNase from the dairy yeast Kluyveromyces lactis, but also translational antibiotics, most pronouncedly hygromycin B (HygB). Specifically, DNA repair mechanisms BER (base excision repair), HR (homologous recombination) and PRR (post replication repair) provided protection, whereas NHEJ (non-homologous end-joining) aggravated toxicity of all translational inhibitors. Analysis of specific BER mutants disclosed a strong HygB, zymocin and PaT protective effect of the endonucleases acting on apurinic sites. In cells defective in AP endonucleases, inactivation of the DNA glycosylase Ung1 increased tolerance to ACNases and HygB. In addition, Mag1 specifically contributes to the repair of DNA lesions caused by HygB. Consistent with DNA damage provoked by translation inhibitors, mutation frequencies were elevated upon exposure to both fungal ACNases and HygB. Since polymerase ζ contributed to toxicity in all instances, error-prone lesion-bypass probably accounts for the mutagenic effects. The finding that differently acting inhibitors of protein biosynthesis induce alike cellular responses in DNA repair mutants is novel and suggests the dependency of genome stability on translational fidelity.

  11. DNA base excision repair of uracil residues in reconstituted nucleosome core particles

    PubMed Central

    Nilsen, Hilde; Lindahl, Tomas; Verreault, Alain

    2002-01-01

    The human base excision repair machinery must locate and repair DNA base damage present in chromatin, of which the nucleosome core particle is the basic repeating unit. Here, we have utilized fragments of the Lytechinus variegatus 5S rRNA gene containing site-specific U:A base pairs to investigate the base excision repair pathway in reconstituted nucleosome core particles in vitro. The human uracil-DNA glycosylases, UNG2 and SMUG1, were able to remove uracil from nucleosomes. Efficiency of uracil excision from nucleosomes was reduced 3- to 9-fold when compared with naked DNA, and was essentially uniform along the length of the DNA substrate irrespective of rotational position on the core particle. Furthermore, we demonstrate that the excision repair pathway of an abasic site can be reconstituted on core particles using the known repair enzymes, AP-endonuclease 1, DNA polymerase β and DNA ligase III. Thus, base excision repair can proceed in nucleosome core particles in vitro, but the repair efficiency is limited by the reduced activity of the uracil-DNA glycosylases and DNA polymerase β on nucleosome cores. PMID:12411511

  12. Measuring the global information society – explaining digital inequality by economic level and education standard

    NASA Astrophysics Data System (ADS)

    Ünver, H.

    2017-02-01

    A main focus of this research paper is to investigate on the explanation of the ‘digital inequality’ or ‘digital divide’ by economic level and education standard of about 150 countries worldwide. Inequality regarding GDP per capita, literacy and the so-called UN Education Index seem to be important factors affecting ICT usage, in particular Internet penetration, mobile phone usage and also mobile Internet services. Empirical methods and (multivariate) regression analysis with linear and non-linear functions are useful methods to measure some crucial factors of a country or culture towards becoming information and knowledge based society. Overall, the study concludes that the convergence regarding ICT usage proceeds worldwide faster than the convergence in terms of economic wealth and education in general. The results based on a large data analysis show that the digital divide is declining over more than a decade between 2000 and 2013, since more people worldwide use mobile phones and the Internet. But a high digital inequality explained to a significant extent by the functional relation between technology penetration rates, education level and average income still exists. Furthermore it supports the actions of countries at UN/G20/OECD level for providing ICT access to all people for a more balanced world in context of sustainable development by postulating that policymakers need to promote comprehensive education worldwide by means of using ICT.

  13. Correlated Mutation in the Evolution of Catalysis in Uracil DNA Glycosylase Superfamily

    PubMed Central

    Xia, Bo; Liu, Yinling; Guevara, Jose; Li, Jing; Jilich, Celeste; Yang, Ye; Wang, Liangjiang; Dominy, Brian N.; Cao, Weiguo

    2017-01-01

    Enzymes in Uracil DNA glycosylase (UDG) superfamily are essential for the removal of uracil. Family 4 UDGa is a robust uracil DNA glycosylase that only acts on double-stranded and single-stranded uracil-containing DNA. Based on mutational, kinetic and modeling analyses, a catalytic mechanism involving leaving group stabilization by H155 in motif 2 and water coordination by N89 in motif 3 is proposed. Mutual Information analysis identifies a complexed correlated mutation network including a strong correlation in the EG doublet in motif 1 of family 4 UDGa and in the QD doublet in motif 1 of family 1 UNG. Conversion of EG doublet in family 4 Thermus thermophilus UDGa to QD doublet increases the catalytic efficiency by over one hundred-fold and seventeen-fold over the E41Q and G42D single mutation, respectively, rectifying the strong correlation in the doublet. Molecular dynamics simulations suggest that the correlated mutations in the doublet in motif 1 position the catalytic H155 in motif 2 to stabilize the leaving uracilate anion. The integrated approach has important implications in studying enzyme evolution and protein structure and function.

  14. Posttraumatic responses to the July 22, 2011 Oslo Terror among Norwegian high school students.

    PubMed

    Nordanger, Dag Ø; Hysing, Mari; Posserud, Maj-Britt; Lundervold, Astri Johansen; Jakobsen, Reidar; Olff, Miranda; Stormark, Kjell Morten

    2013-12-01

    The July 22, 2011, Oslo Terror was defined as a national disaster. Former studies on terror attacks and mass shootings have shown elevated levels of posttraumatic complaints both in direct victims and in general populations. Little is known about how such extreme events in a generally safe society such as Norway would affect an adolescent population. This study examines posttraumatic stress reactions and changes in worldview in relationship to risk factors among 10,220 high school students using data from the ung@hordaland survey. One out of 5 respondents knew someone directly exposed, 55.7% experienced the events to some extent as threatening to their own or their close ones' lives, and 79.9% reported their worldview to be changed. For posttraumatic stress disorder (PTSD) DSM IV criteria, 0.8% reported substantial symptoms of reexperiencing (Criterion B), 4.9% of avoidance (Criterion C), and 1.1% of hyperarousal (Criterion D). Greater personal proximity to the events, higher levels of perceived life threat, and being a female or an immigrant predicted higher levels of PTSD symptom distress. Results indicate that the terror events made a deep impression on Norwegian adolescents, but without causing markedly elevated levels of PTSD symptomatology in the general young population.

  15. Putative modifier genes in mevalonate kinase deficiency.

    PubMed

    Marcuzzi, Annalisa; Vozzi, Diego; Girardelli, Martina; Tricarico, Paola Maura; Knowles, Alessandra; Crovella, Sergio; Vuch, Josef; Tommasini, Alberto; Piscianz, Elisa; Bianco, Anna Monica

    2016-04-01

    Mevalonate kinase deficiency (MKD) is an autosomal recessive auto‑inflammatory disease, caused by impairment of the mevalonate pathway. Although the molecular mechanism remains to be elucidated, there is clinical evidence suggesting that other regulatory genes may be involved in determining the phenotype. The identification of novel target genes may explain non‑homogeneous genotype‑phenotype correlations, and provide evidence in support of the hypothesis that novel regulatory genes predispose or amplify deregulation of the mevalonate pathway in this orphan disease. In the present study, DNA samples were obtained from five patients with MKD, which were then analyzed using whole exome sequencing. A missense variation in the PEX11γ gene was observed in homozygosis in P2, possibly correlating with visual blurring. The UNG rare gene variant was detected in homozygosis in P5, without correlating with a specific clinical phenotype. A number of other variants were found in the five analyzed DNA samples from the MKD patients, however no correlation with the phenotype was established. The results of the presents study suggested that further analysis, using next generation sequencing approaches, is required on a larger sample size of patients with MKD, who share the same MVK mutations and exhibit 'extreme' clinical phenotypes. As MVK mutations may be associated with MKD, the identification of specific modifier genes may assist in providing an earlier diagnosis.

  16. Th17-type cytokines, IL-6 and TNF-α synergistically activate STAT3 and NF-kB to promote colorectal cancer cell growth.

    PubMed

    De Simone, V; Franzè, E; Ronchetti, G; Colantoni, A; Fantini, M C; Di Fusco, D; Sica, G S; Sileri, P; MacDonald, T T; Pallone, F; Monteleone, G; Stolfi, C

    2015-07-01

    Colorectal cancers (CRCs) often show a dense infiltrate of cytokine-producing immune/inflammatory cells. The exact contribution of each immune cell subset and cytokine in the activation of the intracellular pathways sustaining CRC cell growth is not understood. Herein, we isolate tumor-infiltrating leukocytes (TILs) and lamina propria mononuclear cells (LPMCs) from the tumor area and the macroscopically unaffected, adjacent, colonic mucosa of patients who underwent resection for sporadic CRC and show that the culture supernatants of TILs, but not of LPMCs, potently enhance the growth of human CRC cell lines through the activation of the oncogenic transcription factors signal transducer and activator of transcription 3 (STAT3) and nuclear factor-kappa B (NF-kB). Characterization of immune cell complexity of TILs and LPMCs reveals no differences in the percentages of T cells, natural killer T cells, natural killer (NK) cells, macrophages and B cells. However, T cells from TILs show a functional switch compared with those from LPMCs to produce large amounts of T helper type 17 (Th17)-related cytokines (that is, interleukin-17A (IL-17A), IL-17F, IL-21 and IL-22), tumor necrosis factor-α (TNF-α) and IL-6. Individual neutralization of IL-17A, IL-17F, IL-21, IL-22, TNF-α or IL-6 does not change TIL-derived supernatant-driven STAT3 and NF-kB activation, as well as their proproliferative effect in CRC cells. In contrast, simultaneous neutralization of both IL-17A and TNF-α, which abrogates NF-kB signaling, and IL-22 and IL-6, which abrogates STAT3 signaling, reduces the mitogenic effect of supernatants in CRC cells. IL-17A, IL-21, IL-22, TNF-α and IL-6 are also produced in excess in the early colonic lesions in a mouse model of sporadic CRC, associated with enhanced STAT3/NF-kB activation. Mice therapeutically given BP-1-102, an orally bioavailable compound targeting STAT3/NF-kB activation and cross-talk, exhibit reduced colon tumorigenesis and diminished expression of

  17. Tumor-associated immune factors are associated with recurrence and metastasis in non-small cell lung cancer

    PubMed Central

    Yan, X; Jiao, S-C; Zhang, G-Q; Guan, Y; Wang, J-L

    2017-01-01

    Dynamic interaction between tumor cells and the microenvironment is critical for tumorigenesis, and cancer immunosurveillance plays an important role in the tumor evolution. In some tumors (such as esophageal cancer, pancreatic cancer and colorectal cancer), studies have shown that the number of tumor-infiltrating lymphocytes (TILs) has a significant relationship with the prognosis, but there is little research on the prognosis of TILs and non-small cell lung cancer (NSCLC) has been performed. Therefore, it is necessary to discover the relationship between the TILs and cytokines with NSCLC prognosis and metastasis in patients. Tumor samples were carefully examined for tissue preservation and complete follow-up. A total of 107 tumor samples from NSCLC patients with radical surgical resection were enrolled for the analysis. All samples were subjected to immunohistochemistry for detection of CD3, CD4, CD8, CD28, forkhead box protein P3 (Foxp3), cytotoxic T lymphocyte-associated protein-4, cyclooxygenase2 (COX-2), transforming growth factor β 1, interleukin-2 (IL-2), interleukin-6, interleukin-10, interleukin-12 receptor and hypoxia inducible factor 1a (HIF-1a). The number, function and location of the targets were analyzed to determine their correlation with disease-free survival (DFS) and overall survival (OS). Immunhistochemical results from 107 samples indicated that the FoxP3+ regulatory TIL (HR=1.336, P=0.031), IL-2 (HR=0.595, P=0.007) and HIF-1a (HR=1.510, P=0.002) levels in tumor cells closely correlated with DFS in a COX analysis model. FoxP3+ regulatory TILs (HR=1.566, P=0.002) significantly correlated with OS and tumor node metastasis staging. The patients were divided into two groups due to the coexpression pattern of the IL-2, FoxP3+ and HIF-1a. The high-risk group had an overall worse survival than those at low risk. We confirmed that Foxp3 expression in lymphocyte and IL-2 expression in tumor cells were associated with recurrence or transfer

  18. Squamous cell carcinomas escape immune surveillance via inducing chronic activation and exhaustion of CD8+ T Cells co-expressing PD-1 and LAG-3 inhibitory receptors

    PubMed Central

    Mishra, Ameet K.; Kadoishi, Tanya; Wang, Xiaoguang; Driver, Emily; Chen, Zhangguo; Wang, Xiao-Jing; Wang, Jing H.

    2016-01-01

    Squamous cell carcinoma (SCC) is the second commonest type of skin cancer. Moreover, about 90% of head and neck cancers are SCCs. SCCs develop at a significantly higher rate under chronic immunosuppressive conditions, implicating a role of immune surveillance in controlling SCCs. It remains largely unknown how SCCs evade immune recognition. Here, we established a mouse model by injecting tumor cells derived from primary SCCs harboring KrasG12D mutation and Smad4 deletion into wild-type (wt) or CD8−/− recipients. We found comparable tumor growth between wt and CD8−/− recipients, indicating a complete escape of CD8+ T cell-mediated anti-tumor responses by these SCCs. Mechanistically, CD8+ T cells apparently were not defective in infiltrating tumors given their relatively increased percentage among tumor infiltrating lymphocytes (TILs). CD8+ TILs exhibited phenotypes of chronic activation and exhaustion, including overexpression of activation markers, co-expression of programmed cell death 1 (PD-1) and lymphocyte activation gene-3 (LAG-3), as well as TCRβ downregulation. Among CD4+ TILs, T regulatory cells (Tregs) were preferentially expanded. Contradictory to prior findings in melanoma, Treg expansion was independent of CD8+ T cells in our SCC model. Unexpectedly, CD8+ T cells were required for promoting NK cell infiltration within SCCs. Furthermore, we uncovered AKT-dependent lymphocyte-induced PD-L1 upregulation on SCCs, which was contributed greatly by combinatorial effects of CD8+ T and NK cells. Lastly, dual blockade of PD-1 and LAG-3 inhibited the tumor growth of SCCs. Thus, our findings identify novel immune evasion mechanisms of SCCs and suggest that immunosuppressive mechanisms operate in a cancer-type specific and context-dependent manner. PMID:27835902

  19. PD-1/SHP-2 inhibits Tc1/Th1 phenotypic responses and the activation of T cells in the tumor microenvironment.

    PubMed

    Li, Jing; Jie, Hyun-Bae; Lei, Yu; Gildener-Leapman, Neil; Trivedi, Sumita; Green, Tony; Kane, Lawrence P; Ferris, Robert L

    2015-02-01

    Immune rejection of tumors is mediated by IFNγ production and T-cell cytolytic activity. These processes are impeded by PD-1, a coinhibitory molecule expressed on T cells that is elevated in tumor-infiltrating lymphocytes (TIL). PD-1 elevation may reflect T-cell exhaustion marked by decreased proliferation, production of type I cytokines, and poor cytolytic activity. Although anti-PD-1 antibodies enhance IFNγ secretion after stimulation of the T-cell receptor (TCR), the mechanistic link between PD-1 and its effects on T-cell help (Tc1/Th1 skewing) remains unclear. In prospectively collected cancer tissues, we found that TIL exhibited dampened Tc1/Th1 skewing and activation compared with peripheral blood lymphocytes (PBL). When PD-1 bound its ligand PD-L1, we observed a marked suppression of critical TCR target genes and Th1 cytokines. Conversely, PD-1 blockade reversed these suppressive effects of PD-1:PD-L1 ligation. We also found that the TCR-regulated phosphatase SHP-2 was expressed higher in TIL than in PBL, tightly correlating with PD-1 expression and negative regulation of TCR target genes. Overall, these results defined a PD-1/SHP-2/STAT1/T-bet signaling axis mediating the suppressive effects of PD-1 on Th1 immunity at tumor sites. Our findings argue that PD-1 or SHP-2 blockade will be sufficient to restore robust Th1 immunity and T-cell activation and thereby reverse immunosuppression in the tumor microenvironment.

  20. Fast SiPM Readout of the PANDA TOF Detector

    NASA Astrophysics Data System (ADS)

    Böhm, M.; Lehmann, A.; Motz, S.; Uhlig, F.

    2016-05-01

    For the identification of low momentum charged particles and for event timing purposes a barrel Time-of-Flight (TOF) detector surrounding the interaction point is planned for the PANDA experiment at FAIR . Since the boundary conditions in terms of available radial space and radiation length are quite strict the favored layout is a hodoscope composed of several thousand small scintillating tiles (SciTils) read out by silicon photomultipliers (SiPMs). A time resolution of well below 100 ps is aimed for. With the originally proposed 30 × 30 × 5 mm3 SciTils read out by two single 3 × 3 mm2 SiPMs at the rims of the scintillator the targeted time resolution can be just reached, but with a considerable position dependence across the scintillator surface. In this paper we discuss other design options to further improve the time resolution and its homogeneity. It will be shown that wide scintillating rods (SciRods) with a size of, e.g., 50 × 30 × 5 mm3 or longer and read out at opposite sides by a chain of four serially connected SiPMs a time resolution down to 50 ps can be reached without problems. In addition, the position dependence of the time resolution is negligible. These SciRods were tested in the laboratory with electrons of a 90Sr source and under real experimental conditions in a particle beam at CERN. The measured time resolutions using fast BC418 or BC420 plastic scintillators wrapped in aluminum foil were consistently between 45 and 75 ps dependent on the SciRod design. This is a significant improvement compared to the original SciTil layout.

  1. Immunoscore encompassing CD3+ and CD8+ T cell densities in distant metastasis is a robust prognostic marker for advanced colorectal cancer

    PubMed Central

    Kwak, Yoonjin; Koh, Jiwon; Kim, Duck-Woo; Kang, Sung-Bum; Kim, Woo Ho; Lee, Hye Seung

    2016-01-01

    Background The immunoscore (IS), an index based on the density of CD3+ and CD8+ tumor-infiltrating lymphocytes (TILs) in the tumor center (CT) and invasive margin (IM), has gained considerable attention as a prognostic marker. Tumor-associated macrophages (TAMs) have also been reported to have prognostic value. However, its clinical significance has not been fully clarified in patients with advanced CRC who present with distant metastases. Methods The density of CD3+, CD4+, CD8+, FOXP3+, CD68+, and CD163+ immune cells within CRC tissue procured from three sites–the primary CT, IM, and distant metastasis (DM)–was determined using immunohistochemistry and digital image analyzer (n=196). The IS was obtained by quantifying the densities of CD3+ and CD8+ TILs in the CT and IM. IS-metastatic and IS-macrophage–additional IS models designed in this study–were obtained by adding the score of CD3 and CD8 in DM and the score of CD163 in primary tumors (CT and IM), respectively, to the IS. Result Higher IS, IS-metastatic, and IS-macrophage values were significantly correlated with better prognosis (p=0.020, p≤0.001, and p=0.005, respectively). Multivariate analysis revealed that only IS-metastatic was an independent prognostic marker (p=0.012). No significant correlation was observed between KRAS mutation and three IS models. However, in the subgroup analysis, IS-metastatic showed a prognostic association regardless of the KRAS mutational status. Conclusion IS is a reproducible method for predicting the survival of patients with advanced CRC. Additionally, an IS including the CD3+ and CD8+ TIL densities at DM could be a strong prognostic marker for advanced CRC. PMID:27835889

  2. T cell receptor transgenic lymphocytes infiltrating murine tumors are not induced to express foxp3

    PubMed Central

    2011-01-01

    Regulatory T cells (Treg) that express the transcription factor Foxp3 are enriched within a broad range of murine and human solid tumors. The ontogeny of these Foxp3 Tregs - selective accumulation or proliferation of natural thymus-derived Treg (nTreg) or induced Treg (iTreg) converted in the periphery from naïve T cells - is not known. We used several strains of mice in which Foxp3 and EGFP are coordinately expressed to address this issue. We confirmed that Foxp3-positive CD4 T cells are enriched among tumor-infiltrating lymphocytes (TIL) and splenocytes (SPL) in B16 murine melanoma-bearing C57BL/6 Foxp3EGFP mice. OT-II Foxp3EGFP mice are essentially devoid of nTreg, having transgenic CD4 T cells that recognize a class II-restricted epitope derived from ovalbumin; Foxp3 expression could not be detected in TIL or SPL in these mice when implanted with ovalbumin-transfected B16 tumor (B16-OVA). Likewise, TIL isolated from B16 tumors implanted in Pmel-1 Foxp3EGFP mice, whose CD8 T cells recognize a class I-restricted gp100 epitope, were not induced to express Foxp3. All of these T cell populations - wild-type CD4, pmel CD8 and OTII CD4 - could be induced in vitro to express Foxp3 by engagement of their T cell receptor (TCR) and exposure to transforming growth factor β (TGFβ). B16 melanoma produces TGFβ and both pmel CD8 and OTII CD4 express TCR that should be engaged within B16 and B16-OVA respectively. Thus, CD8 and CD4 transgenic T cells in these animal models failed to undergo peripheral induction of Foxp3 in a tumor microenvironment. PMID:22112546

  3. Evaluation of γ-retroviral vectors that mediate the inducible expression of IL-12 for clinical application.

    PubMed

    Zhang, Ling; Feldman, Steven A; Zheng, Zhili; Chinnasamy, Nachimuthu; Xu, Hui; Nahvi, Azam V; Dudley, Mark E; Rosenberg, Steven A; Morgan, Richard A

    2012-06-01

    The clinical application of interleukin-12 (IL-12) has been hindered by the toxicity associated with its systemic administration. To potentially overcome this problem, we developed a promoter designed to direct IL-12 expression within the tumor environment using an inducible composite promoter containing binding motifs for the nuclear factor of activated T cells (NFAT) linked to a minimal IL-2 promoter. In this study, the NFAT promoter was coupled to a single-chain human IL-12 gene and inserted into 2 γ-retroviral self-inactivating vectors (SERS.NFAT.hIL12 and SERS.NFAT.hIL12.PA2) and 1 γ-retroviral vector (MSGV1.NFAT.hIL.12 PA2). Peripheral blood lymphocytes (PBLs) were double transduced with an antigen-specific T-cell receptor and the 3 NFAT.hIL12 vectors. Evaluation of inducible IL-12 expression, transduction efficiency, and vector production considerations led to the choice of the MSGV1.NFAT.hIL12.PA2 vector for clinical application. MSGV1.NFAT.hIL12.PA2 PG13 retroviral vector producer cell clones were screened by transduction of tumor antigen-specific PBLs. On the basis of expression studies in PBL, clone D3 was chosen to produce clinical-grade viral vector supernatant and was demonstrated to efficiently transduce young tumor-infiltrating lymphocytes (TIL). The vector-transduced young TIL with known tumor recognition demonstrated specific inducible IL-12 production after coculture with HLA-matched tumor targets and had augmented effector function as demonstrated by increased IFN-γ secretion. These results support the clinical application of adoptive transfer of young TIL engineered with the NFAT.hIL12 vector as a new approach for cancer immunotherapy.

  4. Squamous cell carcinomas escape immune surveillance via inducing chronic activation and exhaustion of CD8+ T Cells co-expressing PD-1 and LAG-3 inhibitory receptors.

    PubMed

    Mishra, Ameet K; Kadoishi, Tanya; Wang, Xiaoguang; Driver, Emily; Chen, Zhangguo; Wang, Xiao-Jing; Wang, Jing H

    2016-12-06

    Squamous cell carcinoma (SCC) is the second commonest type of skin cancer. Moreover, about 90% of head and neck cancers are SCCs. SCCs develop at a significantly higher rate under chronic immunosuppressive conditions, implicating a role of immune surveillance in controlling SCCs. It remains largely unknown how SCCs evade immune recognition. Here, we established a mouse model by injecting tumor cells derived from primary SCCs harboring KrasG12D mutation and Smad4 deletion into wild-type (wt) or CD8-/- recipients. We found comparable tumor growth between wt and CD8-/- recipients, indicating a complete escape of CD8+ T cell-mediated anti-tumor responses by these SCCs. Mechanistically, CD8+ T cells apparently were not defective in infiltrating tumors given their relatively increased percentage among tumor infiltrating lymphocytes (TILs). CD8+ TILs exhibited phenotypes of chronic activation and exhaustion, including overexpression of activation markers, co-expression of programmed cell death 1 (PD-1) and lymphocyte activation gene-3 (LAG-3), as well as TCRβ downregulation. Among CD4+ TILs, T regulatory cells (Tregs) were preferentially expanded. Contradictory to prior findings in melanoma, Treg expansion was independent of CD8+ T cells in our SCC model. Unexpectedly, CD8+ T cells were required for promoting NK cell infiltration within SCCs. Furthermore, we uncovered AKT-dependent lymphocyte-induced PD-L1 upregulation on SCCs, which was contributed greatly by combinatorial effects of CD8+ T and NK cells. Lastly, dual blockade of PD-1 and LAG-3 inhibited the tumor growth of SCCs. Thus, our findings identify novel immune evasion mechanisms of SCCs and suggest that immunosuppressive mechanisms operate in a cancer-type specific and context-dependent manner.

  5. Groundwater Productivity and Quality of The Quartzite Ridge of RÓdA~o and Their Vicinities (center of Portugal)

    NASA Astrophysics Data System (ADS)

    Duque, J.; Chambel, A.

    When the ESA Huygens Probe arrives at Titan in 2005, measurements taken during and after the descent through the atmosphere are likely to revolutionise our under- standing of SaturnSs most enigmatic moon. The accurate atmospheric profiling of Titan from these measurements will require knowledge of the probe descent trajectory and in some cases attitude history, whilst certain atmospheric information (e.g. wind speeds) may be inferred directly from the probe dynamics during descent. Two of the instruments identified as contributing valuable information for the reconstruction of the probeSs parachute descent dynamics are the Surface Science Package Tilt sensor (SSP-TIL) and the Huygens Atmospheric Structure Instrument servo accelerometer (HASI-ACC). This presentation provides an overview of these sensors and their static calibration before describing an investigation into their real-life dynamic performance under simulated Titan-gravity conditions via a low-cost parabolic flight opportunity. The combined use of SSP-TIL and HASI-ACC in characterising the aircraft dynam- ics is also demonstrated and some important challenges are highlighted. Results from some simple spin tests are also presented. Finally, having validated the performance of the sensors under simulated Titan conditions, estimates are made as to the output of SSP-TIL and HASI-ACC under a variety of probe dynamics, ranging from verti- cal descent with spin to a simple 3 degree-of-freedom parachute descent model with horizontal gusting. It is shown how careful consideration must be given to the instru- mentsS principles of operation in each case, and also the impact of the sampling rates and resolutions as selected for the Huygens mission. The presentation concludes with a discussion of ongoing work on more advanced descent modelling and surface dy- namics modelling, and also of a proposal for the testing of the sensors on a sea-surface.

  6. Reduced-gravity Testing of The Huygens Probe Ssp Tiltmeter and Hasi Accelerometer Sensors and Their Role In Reconstruction of The Probe Descent Dynamics

    NASA Astrophysics Data System (ADS)

    Ghafoor, N.; Zarnecki, J.

    When the ESA Huygens Probe arrives at Titan in 2005, measurements taken during and after the descent through the atmosphere are likely to revolutionise our under- standing of SaturnSs most enigmatic moon. The accurate atmospheric profiling of Titan from these measurements will require knowledge of the probe descent trajectory and in some cases attitude history, whilst certain atmospheric information (e.g. wind speeds) may be inferred directly from the probe dynamics during descent. Two of the instruments identified as contributing valuable information for the reconstruction of the probeSs parachute descent dynamics are the Surface Science Package Tilt sensor (SSP-TIL) and the Huygens Atmospheric Structure Instrument servo accelerometer (HASI-ACC). This presentation provides an overview of these sensors and their static calibration before describing an investigation into their real-life dynamic performance under simulated Titan-gravity conditions via a low-cost parabolic flight opportunity. The combined use of SSP-TIL and HASI-ACC in characterising the aircraft dynam- ics is also demonstrated and some important challenges are highlighted. Results from some simple spin tests are also presented. Finally, having validated the performance of the sensors under simulated Titan conditions, estimates are made as to the output of SSP-TIL and HASI-ACC under a variety of probe dynamics, ranging from verti- cal descent with spin to a simple 3 degree-of-freedom parachute descent model with horizontal gusting. It is shown how careful consideration must be given to the instru- mentsS principles of operation in each case, and also the impact of the sampling rates and resolutions as selected for the Huygens mission. The presentation concludes with a discussion of ongoing work on more advanced descent modelling and surface dy- namics modelling, and also of a proposal for the testing of the sensors on a sea-surface.

  7. Temporal and spatial discordance of programmed cell death-ligand 1 expression and lymphocyte tumor infiltration between paired primary lesions and brain metastases in lung cancer

    PubMed Central

    Mansfield, A. S.; Aubry, M. C.; Moser, J. C.; Harrington, S. M.; Dronca, R. S.; Park, S. S.; Dong, H.

    2016-01-01

    Background The dynamics of PD-L1 expression may limit its use as a tissue-based predictive biomarker. We sought to expand our understanding of the dynamics of PD-L1 expression and tumor-infiltrating lymphocytes (TILs) in patients with lung cancer-related brain metastases. Experimental design Paired primary lung cancers and brain metastases were identified and assessed for PD-L1 and CD3 expression by immunohistochemistry. Lesions with 5% or greater PD-L1 expression were considered positive. Agreement statistics and the χ2 or Fisher's exact test were used for analysis. Results We analyzed 146 paired lesions from 73 cases. There was disagreement of tumor cell PD-L1 expression in 10 cases (14%, κ = 0.71), and disagreement of TIL PD-L1 expression in 19 cases (26%, κ = 0.38). Most paired lesions with discordant tumor cell expression of PD-L1 were obtained 6 or more months apart. When specimens were categorized using a proposed tumor microenvironment categorization scheme based on PD-L1 expression and TILs, there were significant changes in the classifications because many of the brain metastases lacked either PD-L1 expression, tumor lymphocyte infiltration or both even when they were present in the primary lung cancer specimens (P = 0.009). Conclusions We identified that there are significant differences between the tumor microenvironment of paired primary lung cancers and brain metastases. When physicians decide to treat patients with lung cancer with a PD-1 or PD-L1 inhibitor, they must do so in the context of the spatial and temporal heterogeneity of the tumor microenvironment. PMID:27502709

  8. Ada Compiler Validation Summary Report: Certificate Number: 940325S1. 11344 DDC-I, DACS Sun SPARC/Solaris to 80186 Bare Ada Cross Compiler System with Rate Monotonic Scheduling, Version 4.6.4 Sun SPARCclassic => Intel iSBC 186/100 (Bare Machine)

    DTIC Science & Technology

    1994-03-25

    5*n" "i~m~ &4" W4000 Vmh~ . a=. 1=3-EOTTP AND KIE 4. TIL5N . FUNDING 940325SI.11344, AMF 94ddc500_iC DDC-I, DACS Sun SPARC/Solaris to 80186 Bare Ada...DISTRIBUTION Approved for Public Release; .. distribution unlimited 13. (Maxinxm 200 Host: Sun SPARCclassic (under Sokaris, Release 2.1) Target: Intel...COMPILER VALIDATION SUMMARY REPORT: Certificate Number: 940325S1.11344 DDC-I DACS Sun SPARC/Solaris to 80186 Bare Ada Cross Compiler System with Rate

  9. 1. Innovative Relaxor-Based PiezoCrystals: Phase Diagrams, Crystal Growth, Domain Structures and Electric Properties. 2. Piezo- and Ferroelectric Materials Based on Morphotropic Phase Boundary Synthesis, Characterization and Structure - Property Relations

    DTIC Science & Technology

    2006-03-31

    Aug. 2-8, 2003. [48] Z.-G. Ye, A. Bokov and M . Maglione , "Dielectric and Structural Properties of Relaxor Ferroelectrics", 55 Years of Ferroelectrics...Pacific Center for Advanced Materials and Microstructure (PCAMM), NRC / UBC, Dec. 4, 2004. [64] A. A. Bokov, M . Maglione and Z.-G. Ye, "Low-Frequency...Argentina, Sept. 5 - 9, 2005. [78] A. A. Bokov, M . Maglione and Z.-G. Ye, "Critical Dielectric Behaviour in Ba(Til-,Zr,)0 3 Solid Solution" (0), 1 1 1h

  10. Air Force Human Resources Laboratory Annual Report - Fiscal Year 1981.

    DTIC Science & Technology

    1982-06-01

    with its application of fiber-optics. technical reviews was based on a week-long meeting with helmet- mounted display. advanced (IG. and division...optics. training and combat conditions. The thrust consists of helmet- mounted display. advanced computer image four subthtrusts: (a) Air Combat ,’aining...formais of Mntissiort T rainier . N\\e% efforts inivolvi ig t[lie use of T \\C expjetiritenrtalI cotoli are- di fficulIt a ti(l % here resources toi BR A L

  11. Producibility and Serviceability of Kevlar-49 Structures Made on Hot Layup Tools

    DTIC Science & Technology

    1975-05-01

    34 roducibility rviceability evlar-4 ructures de t yup ols. " e poses s gram re: • • aluate duc bility licopter ctural ponent de m evlar-4...ore tensiv e pera ture urvey ver e LT urface rea . ut or is ro gram airings e r e bjected ur e e f pr oximate l y inutes nsur e at cured rea s...9 la inates 2 1 Go od Fairly clean edges c: ɘ Fu ller ton Fib e rg las s ::T ~ outer B it til :I: N outi ng Techno log y As ooc. 9 l aminate s

  12. Investigation and Assessment of Hardware for Guidance of Gliding Airdrop Systems

    DTIC Science & Technology

    1979-02-28

    i r- i" r- 1 u VI ?x U 41 IS u u ai 2 *J 41 u m " > 4i i— ** 4i a 4 til a i- c E «<— a. x U 41 lii u 41 ’F— ftoC IS 5N *« £5 V...and these range measurements are translated through tri-lateration into a position fix. The family trees of this configuration ai -e shown in...Configuration description Estimated cost System cost 4 airborne + 1 ground Ai rborne Ground I II III IV V _______ DME/Airborne DF DME/Ground ADF Mini

  13. Refractive Index of Silicon and Germanium and Its Wavelength and Temperature Derivatives.

    DTIC Science & Technology

    1979-03-01

    59. Tauc , J., Abraham, A., Pajasova, L., Grigorovici, R., and Vancu, A., "Optical Properties of Non-Crystalline Semiconductors," in Physics of Non...259 onm 1 84 9!. vojn: .p r us TlI II io L, rinc 11 1US I11L ni . 3 1.98’ ,iii til n i,:n ;Ivvi i Li o i Ili LtI loi ,I ,.kii d.i / T ’I 2 .1) 11 L...index measured using interference method. 40 (T-298 K) Amorphous germanium thin film prepared Tauc , J., Abraham, A., 159) 0.695 4.742 by evaporation

  14. Exploration and Production of Hydrocarbon Resources in Coastal Alabama and Mississippi. Executive Summary.

    DTIC Science & Technology

    1984-11-01

    service industry Increases at meeting more than l /2 Iold workecs; most coald maintenance. sioon In Tanlrs 2-, i region, so increases points. Potential...hydrocarbon Industry is developing In the study area. included in this minor impacts category are: 13-3 r I ct, c 1 til b l tv; rom .a A 11 u I and p...537-2o58 iv HAER 4 - MITIGATING MEASURES4- I NTdO DUCT ION 4-1 REGUL.AT RY RFUIREHENTrs 4-1 INDUSTRY PRACrIC] ci4-1 5V’[lcR P~OTENTIAL, MITIGATING

  15. Seal Technology in Gas Turbine Engines

    DTIC Science & Technology

    1978-08-01

    B) Specialists’ Meeting In London, United Klngdozn 6-7 April 1978. TIlE MISSION OF AGARD The mission of AGARD is to bring together the leading...consisting of officially appointed senior representatives from each member nation. The mi •ion of AGARD Is carried out through the Panels which are...teclanologio des Joints de turbines A giz. L 𔄀tude flat llmitde aux cia oil intervient un mouvement relatif entre let divergse partle des Joints, et couvrit i Ia

  16. National Hydroelectric Power Resources Study: Regional Assessment: Volume XXII: Western Systems Coordinating Council, (WSCC). Volume 22

    DTIC Science & Technology

    1981-09-01

    I 5 _ BINGHAM -* - -• I05NPWOOgS * HAKER CREEK - I 5 _ BLAINE * * -• _ 43 24.0 - SNAKE RIVER _ 112 Q.o_ • • - - - NW . - Til _ * * EASLEY HUT...0 * 7~S. o. NNYUVUN * • I 5 • HAKER BURNT RIVER * 117 39.9 * IS • 8100 * 7bO • 303b. 245.33 • • * NW • • b50 * 100.0. 80:9. 7&0 * 303b...IS • 403500 * 7811 * 2585 * 1805.3 * * * NW * * 1021 * 180.0* 339.0 * 7811 * 2585 * * UYVUNNNUN * * * HAKER 11250000

  17. Synthesis, Spectroscopic and Toxicity Studies of Titanocene Chelates of Isatin-3-Thiosemicarbazones

    PubMed Central

    Vatsa, Garima; Pandey, O. P.; Sengupta, S. K.

    2005-01-01

    The reactions of bis(cyclopentadienyl)titanium(IV) dichloride with a new class of thiosemicarbazone (LH2), derived by condensing isatin with different N(4)-substituted thiosemicarbazides, have been studied and products of type [Cp2Ti(L)] have been isolated. On the basis of various physico-chemical and spectral studies, five coordinate structures have been assigned to these derivatives. Toxicity studies of titanocene complexes at tbur different concentrations have been carried out against snail Lymnaea acuminata. The effect of most potent compounds on the activity of acetylcholinesterase enzyme, which inhibits the activity of enzyme, possibly by the formation of enzyme-inhibitor complex, was also studied. PMID:18365096

  18. Military Health Service System Ambulatory Work Unit (AWU).

    DTIC Science & Technology

    1988-04-01

    probability of a value outside 2.0 standard deviations ranges from .0456 for A-3 -Al" 122 NILITRY HEALTH SERVICE SYSTEM AMULRTORY MORKe UurNT CANdU )(U) ARMY...00 2 w w >4 &4~ ~z til 0n0 rz~ co00 P - xU) I : 0 ) ) %z E-4 z U) HHw H~~ ~ ~ 0 olf4 E1E- 004 H U) UDm 0 4 E- H wE- ~ H E-4 H 04 H Z U H Q z HO H % x z

  19. Military Schools for Prospective Officers of the Soviet Armed Forces

    DTIC Science & Technology

    1975-03-31

    SCHOOLS4FOR IROSPECTIVE O’FICERS, OF TilE SOVIET MED FORCES DDC(, 1. ie i E II n - CAPTAIN JA MES- - G RU S ID501UDUTION s81.’ . Approved jot l Divtiumion...Ma~or 2 Gonoral WK ’Kre 01amalycsakia komandirikie -navyki, Voonnyl etlk o (1973), Pp. 57-58. 4 14400 orel N. Ar4litov and~ Captain Y., Ptamolo0...cosmopolitan mision (frequent ’foreign contacts), also are doing well In this field. eCf., Naval Captain L. Ufimtsev and Commander A.- I)t~Aktual’nyt zaclach

  20. The Turbulent Boundary Layer: An Experimental Study of the Transport of Momentum and Heat with the Effect of Roughness

    DTIC Science & Technology

    1975-05-01

    COMMERCE TIlE TURBULENT BOUNDARY LAYER: AN EXPERIMENTAL STUDY OF THE TRANSPORT OF 1 M(IMENTUM AND HEAT W\\ ITHl THE EFFECT OF ROUGIINESS I • • 11. . 1 ...1975 NATIONAL TECHNICAL INFORMATION SERVICE US D.l’. 2 C....51.3’ tt.VA. 22t51 II Unclassified - SECURTY CLAISSIFICATION OF TUIS PAGE 1 --. 0- 1 ...EoI1*’*dl REPORT DOCUMENTATION4 PAGE BEOECMLEIGFR REPO T N] 1 :ER:.10VT ACCESSION NO. 3 RsECIPIENT Sl CATALOG NUMBER 4 TITLE (sI4S.btIIII.) 5. TYPE OF

  1. SANs and Large Scale Data Migration at the NASA Center for Computational Sciences

    NASA Technical Reports Server (NTRS)

    Salmon, Ellen M.

    2004-01-01

    Evolution and migration are a way of life for provisioners of high-performance mass storage systems that serve high-end computers used by climate and Earth and space science researchers: the compute engines come and go, but the data remains. At the NASA Center for Computational Sciences (NCCS), disk and tape SANs are deployed to provide high-speed I/O for the compute engines and the hierarchical storage management systems. Along with gigabit Ethernet, they also enable the NCCS's latest significant migration: the transparent transfer of 300 Til3 of legacy HSM data into the new Sun SAM-QFS cluster.

  2. Development of a Decompression Algorithm for Constant 0.7 ATA Oxygen Partial Pressure in Helium Diving.

    DTIC Science & Technology

    1985-04-01

    HVAL02 140/15 ( 1 ) Hips, costo -vertebral Joints, costo -chondral junctions. 4 Bin Hip pain gone by 5 min at 60 FSW. All other Symptoms gone upon...AD-A158 142 UNCLASSIFIED DEVELOPMENT OF A DECOMPRESSION ALGORITHM FOR CONSTANT 1 /Jt 97 ATA OXVGEN PARTI.. <U> NAVY EXPERIMENTAL DIVING UNIT...PANAMA CITV FL E D THALMANN APR 85 NEDU- 1 -85 F/G 6/19 NL T i i •• » 1.1.1« j*. .*••’ ’*• J ’ ’*"’" -f" ll til5 |M 2-5 •• 2

  3. The Influence of a Lower Heated Tube on Nucleate Pool Boiling from a Horizontal Tube

    DTIC Science & Technology

    1992-06-01

    AD-A256 833 NAVAL POSTGRADUATE SCHOOL Monterey, California (,-A So Ic THESIS TIlE INFLUENCE OF A LOWER IIEATED TUBE ON NUCLEATE POOL BOILING FROM A...HORIZONTAL TUBE by Lannic R. Lake June 1992 Thesis Advisor Paul J. Marto Co-Advisor Stephen B. Memory Approved for public rclcase; distribution is...day) 15 Page • nt Master’s Thesis From . To June 1992 16 Supplementary Notation The views expressed in this thesis are those of the author and do

  4. Classification Management. Journal. Volume X - 1974. Papers from the National Seminar (10th), 16-17-18 July 1974

    DTIC Science & Technology

    1974-01-01

    Contract. Even at the end of the three year period, if a continued need to 1 : - I dl i~ t Ivtop nttaa go sil tlisa Itraltec il~ t onf In ti-l 142. Fiartlivi...This is one of the problems dares, standards and criteria necessary to meet we are continuing to address. In this connection, changing conditions...inforhiation. Tlhe major problems arise diem satlaries. *and so, forth. It doesn ’ t matter when and It peoplIr feel a c tasuif icat ion is made

  5. Erratum to "Predicting sulphur and nitrogen deposition using a simple statistical method" [Atmos. Environ. 140 (2016) 456-468

    NASA Astrophysics Data System (ADS)

    Oulehle, Filip; Kopáček, Jiří; Chuman, Tomáš; Černohous, Vladimír; Hůnová, Iva; Hruška, Jakub; Krám, Pavel; Lachmanová, Zora; Navrátil, Tomáš; Štěpánek, Petr; Tesař, Miroslav; Evans, Christopher D.

    2016-10-01

    The Journal regrets that the author's names were tagged incorrectly resulting in author forenames appearing as surnames. The correct author names are: Filip Oulehle, Jiří Kopáček, Tomáš Chuman, Vladimír Černohous, Iva Hůnová, Jakub Hruška, Pavel Krám, Zora Lachmanová, Tomáš Navrátil, Petr Štěpánek, Miroslav Tesař, Christopher D. Evans. The Journal would like to apologise for any inconvenience caused.

  6. Association Between NRAS and BRAF Mutational Status and Melanoma-Specific Survival Among Patients With Higher Risk Primary Melanoma

    PubMed Central

    Thomas, Nancy E.; Edmiston, Sharon N.; Alexander, Audrey; Groben, Pamela A.; Parrish, Eloise; Kricker, Anne; Armstrong, Bruce K.; Anton-Culver, Hoda; Gruber, Stephen B.; From, Lynn; Busam, Klaus J.; Hao, Honglin; Orlow, Irene; Kanetsky, Peter A.; Luo, Li; Reiner, Anne S.; Paine, Susan; Frank, Jill S.; Bramson, Jennifer I.; Marrett, Lorraine D.; Gallagher, Richard P.; Zanetti, Roberto; Rosso, Stefano; Dwyer, Terence; Cust, Anne E.; Ollila, David W.; Begg, Colin B.; Berwick, Marianne; Conway, Kathleen

    2015-01-01

    Importance NRAS and BRAF mutations in melanoma inform current treatment paradigms but their role in survival from primary melanoma has not been established. Identification of patients at high risk of melanoma-related death based on their primary melanoma characteristics before evidence of recurrence could inform recommendations for patient follow-up and eligibility for adjuvant trials. Objective To determine tumor characteristics and survival from primary melanoma by somatic NRAS and BRAF status. Design, Setting, and Participants A population-based study with median follow-up of 7.6 years for 912 patients with first primary cutaneous melanoma analyzed for NRAS and BRAF mutations diagnosed in the year 2000 from the United States and Australia in the Genes, Environment and Melanoma Study and followed through 2007. Main Outcomes and Measures Tumor characteristics and melanoma-specific survival of primary melanoma by NRAS and BRAF mutational status. Results The melanomas were 13% NRAS+, 30% BRAF+, and 57% with neither NRAS nor BRAF mutation (wildtype). In a multivariable model including clinicopathologic characteristics, NRAS+ melanoma was associated (P<.05) with mitoses, lower tumor infiltrating lymphocyte (TIL) grade, and anatomic site other than scalp/neck and BRAF+ melanoma was associated with younger age, superficial spreading subtype, and mitoses, relative to wildtype melanoma. There was no significant difference in melanoma-specific survival for melanoma harboring mutations in NRAS (HR 1.7, 95% CI, 0.8–3.4) or BRAF (HR, 1.5, 95% CI, 0.8–2.9) compared to wildtype melanoma adjusted for age, sex, site, AJCC tumor stage, TIL grade, and study center. However, melanoma-specific survival was significantly poorer for higher risk (T2b or higher stage) tumors with NRAS (HR 2.9; 95% CI 1.1–7.7) or BRAF (HR 3.1; 95% CI 1.2–8.5) mutations but not for lower risk (T2a or lower) tumors (P=.65) adjusted for age, sex, site, AJCC tumor stage, TIL grade, and study center

  7. Highlights of the society for immunotherapy of cancer (SITC) 27th annual meeting

    PubMed Central

    2013-01-01

    The 27th annual meeting of the Society for Immunotherapy of Cancer (SITC) was held on October 26–28, 2012 in North Bethesda, Maryland and the highlights of the meeting are summarized. The topics covered at this meeting included advances in cancer treatment using adoptive cell therapy (ACT), oncolytic viruses, dendritic cells (DCs), immune check point modulators and combination therapies. Advances in immune editing of cancer, immune modulation by cancer and the tumor microenvironment were also discussed as were advances in single cell analysis and the manufacture and potency testing of tumor infiltrating lymphocytes (TIL).

  8. Cell Transfer Therapy for Cancer: Past, Present, and Future

    PubMed Central

    Qian, Xiaoling; Wang, Xian; Jin, Hongchuan

    2014-01-01

    Cell transfer therapy for cancer has made a rapid progress recently and the immunotherapy has been recognized as the fourth anticancer modality after operation, chemotherapy, and radiotherapy. Lymphocytes used for cell transfer therapy include dendritic cells, natural killer (NK) cells, and T lymphocytes such as tumor-infiltrating lymphocytes (TILs) and cytotoxic T lymphocytes (CTLs). In vitro activated or engineered immune cells can traffic to cancer tissues to elicit persistent antitumor immune response which is very important especially after immunosuppressive treatments such as chemotherapy. In this review, we overviewed recent advances in the exploration of dendritic cells, NK cells, and T cells for the treatment of human cancer cells. PMID:24741604

  9. Optimal Stationary Linear Control of the Wiener Process.

    DTIC Science & Technology

    1980-02-29

    OPTIMAL STATIONARY LINEAR CONTROL OF THE WIENER PROCESS. (U) FES 80 V E BENES. I KARATZAS AFOSR-76-3063 UNCLASSIFIED AFOSR -TR-8O-0338 NL...mElllllllllll mhmmmmmmmmm mmmmmmmr( EFOSR-TR- 2 0-0S8 0 3 OPTIMAL STATIONARY LINEAR CONTROL OF THE WIENER PROCESS by LEVEL CVACLAV E. BENES Bell Telephone...or " DIst. special D U L I. F UNCLASSIFIED _ _ _ , ,_I_ _ _ _ _ _. OPTIMAL STATIONARY LINEAR CONTROL OF TIlE WIENER PROCESS V’aclav E. Benes and

  10. How Quickly Can Attention Affect Form Perception?

    DTIC Science & Technology

    1987-11-01

    of targetA in1c’ntxi i,’jtio iretLrairilg triails giveni pri or to Th is exicr inmen t 4) run ii iei ia to lyj after Fxperim int I, uslnj t;]il same...cue would still require the retrieval of the to-be-attended location from memory . fut there is no longer jiuch reason to suspect that the time...dttention. The Journal of Neuroscience, 4, 1863-1874. Reeves, A., & Sperling, G. (in press). Attentiori rjati, it, sh1 r t term visual memory

  11. 3W Analysis of FY 78 Army Aircraft Accidents.

    DTIC Science & Technology

    1979-05-01

    Callen, J. R., "Pilot Error Accidents Aren’t All Pilot," United States Army Agency for Aviation Safety, May 1975. 25 ~ ~ - APPENDICES 27 til - -1P I If 2...LJSASC-TR-79- ML DIS RII3UMUN STATZLIA USASC TECHNICAL REPORT Approved fox public releaqw TR 79.3 Ana Iyss Of FY78 704Acidents. A.L:fM ;what happened...construed as an official Department of the Army position, policy , or decision, unless so designated by other official authority. The findings of this

  12. The Shock and Vibration Bulletin. Part 2. Ship Shock, Shock and Blast and Ground Shock

    DTIC Science & Technology

    1984-06-01

    1 ~S*ISie spoloo-s TSwSo SWot P1. * Iseel PS.... 0 A*6I 6 r.S 0 " .- 3 *(O f~*,,* m a se aa’atevw b te rd 2Plto oltv ~pems tu tu toe po00til...4--------4 are shown with the arrows indicating Icame go. WIII~jI I4I1~a (ACC) maxis : the direction of decreasing (y’)m x.. In I. Is .0 Its...corresponds to the maxi - is simply repeated using the other input in mum displacement response was also determined. place of Equation 4-1. The damping

  13. Protein Kinase C is a Mediator of Lipopolysaccharide-Induced Vascular Suppression in the Rat Aorta

    DTIC Science & Technology

    1994-01-01

    28). A possible require- 0 ,] ment for multiple LPS-stimulated mediators to full\\ induce300o •"NOS activity in VSM can explain the apparent paradox...A D - A 286 074 )AGE 0704-0pprove IIIo- 1 DC 199 AGENC U EPORT TYPE AND DATES COVERED 1994L N S-TIL Journal article 4. ITE ND UBITES. Fý;.:DlNG...ADDRESS(ES) 110. SAPOENSCOYRINEGO/RMONNITNOR~ING Naval Medical Research and Development Command AEC EOTNME National Naval Medical Center Building 1 , Towver

  14. Staffing Cyber Operations (Briefing Charts)

    DTIC Science & Technology

    2015-05-01

    w hi ch p os iti on s co ns tit ut e D ire ct P ar tic ip at io n in C yb er sp ac e H os til iti es (D P H ) = C yb er sp ac e C om ba ta...un ity  ID A a lte rn at iv e la rg el y su bs tit ut es c iv ili an s fo r e nl is te d pe rs on ne l a nd co nt ra ct or s  M ili ta ry a

  15. Investigations at Site 32 (41EP325), Keystone Dam Project. A Multicomponent Archeological Site in Western El Paso County, Texas.

    DTIC Science & Technology

    1983-03-01

    In studying thoc surface dlstrLft 151c, of Ire-crack’s( rocks, platn maps shoiwiii surf ici features and dispersed ruock densi I (is (by iwelo’.t ,ind...noteI i r tt til 1 -s:uI roucuiJT I I, t fc-. rocks, hut pi t 7, 1 1n:1 Cul no h,1r*1 e dfc lir it A, wi1thI Feat ure t, Feat ure( 14 is ltterjret’l...clear that all vessels represented in the collection are tempered with sand (each specimen was examined using a variable power binocular micro- scope

  16. Atmospheric Transmittance/Radiance: Computer Code LOWTRAN 6. Supplement: Program Listings

    DTIC Science & Technology

    1983-08-01

    Program Listings F. X. KNEIZYS E. P . SHETTLE W. 0. GALLERY J. HK CHETWYND, Jr. <L. W. ABRFU J,. E, A. SELBY Z S. A. CLOUGH R. W. FENN DTIC ELECT 1...Kneizys J. H. Chetwynd, Jr. E. P . Shettle L. W. Abreu , S. A. Clough W.O0. Galler JI.A Selby* R. W. Fenni S. P ;,.IkPORUING ORGANIZATO N;. r AN. ARES...PATH TO TIlE EXTIRATEFUIWSTRIAL SOURCE. 73 EXA B IN LOADIS A ROS I, EXt’IACTION AND P AlW SO •1 TION COEPFFICIFNTS I.FOR THE APPROPU SATE MODEIL AND

  17. Multiaperture Optics.

    DTIC Science & Technology

    1986-03-01

    is collected and gives rise to one point only. The con- sequence is that the location of the image point is determined by the - eye architecture and is...encountered con- sists only of a sequence of rectangles. The picture language used has only one word "rectangle." The information describing the...l i tt i t l 1t 11111 VI11 21 1 illill t 111111t ... . Ill tt Ill ttttttt Ill ii it itlilt I til liii 1 1111 111111 1 t 11111111illl its1 II till Iti

  18. Experiment in Water Dowsing

    DTIC Science & Technology

    1994-09-01

    0)0 "• •,• • -• )• .EEP C .7 7:i EXPERIMENT IN WATER DOWSING Til I"SIS EEDavid 1. (Gaisford. 1). E. AFIT/GEE/F.NS/94S-O I ~~m - DEPARTMENT... WATER DOWSING THESIS David I. Gaisford, P. E. AFIT/GEE/ENS/94S-01 Approved for Public Release; distribution unlimited =TC QUALITY pSP-ECTED 3 S0694-30622...Specils THESIS APPROVAL Student: David I. Gaisford Class: GEE 94S Thesis Title: Experiment in Water Dowsing Defense Date: 11 Aug 94 Committee Name

  19. Department of Defense, Office of Inspector General Semiannual Report to the Congress, 1 April - 30 September 1984.

    DTIC Science & Technology

    1984-11-29

    2i. • .2•.A . ’~ o ****~ " ’ *~~ " .... ............... -77 -7 - 71𔄁" 7.. ..7.. ......-. . x ’ - INTERNAL AUDIT FOLLOWUP ACCOMPUSHMENTS...rcl ioinit’ ti s il th i specifieatitoits and si atidards, interpret itng ti-st resulIts and - ti~l’ts: 40 involved proiilts with i litlenttig i- X ...Rooms in * steadl of* the 14-passe-nger aircraft being sotlicitecl. lu r- Hawaii. Thel( Nav- x rented hotel acc(Inm mdat ions for * hasing three leased

  20. Foulois and the U.S. Army Air Corps 1931-1935,

    DTIC Science & Technology

    1983-01-01

    anl inhibiting agent . "Fhe War 1)1,1111 heri 111 iiii19~ sI󈧞 JIknossledi!d at mutltitude ot’ tasks that military .1 ou ildl pciioini sonie tar...leasin tilIle post \\ iaatit usiti ! tile 1\\Il ii ecuses of ecotiotlnv and War lDepartnm t conisoiidationi. II oosci &J ot na me a ep cnntdmuine tile remli...briefly considered the wartime strength and composition of the GHQ Air Force. The War Plans Division was reconsidering the RED and ORANGE %sar plans

  1. Development and Evaluation of Reproductive and Developmental Toxicity Tests for Assessing the Hazards of Environmental Contaminants

    DTIC Science & Technology

    1997-08-01

    the report. D. METHODLOGY VI length data were colleced and analyzed E*e°*U^ Eglin AFB soil in sealed jars, exposure in Petn dishes q to...ft 0! as Ta- hn a in m *c O u u rn n. -M 43 u m s. n> ft O n o <u l-i U O r3 -M c U a> ’Si s T3 (0 til 43 « o ffl t^ u 01 ca ’’ ’! -M

  2. E15, Cost Analysis and Funding. User’s Manual Version 1.0

    DTIC Science & Technology

    1991-04-01

    C es Ava il amd/o - Dist special CONTRACT DAAA2I-86-D-0025 Ds PC~ for HO US AMCCOM INTEGRATED LOGISTIC SUPPORT OFFICE AMSMC-LSP ROCK ISLAND, IL by...provided by Barbara Boren and Denise Montanez . We gratefully acknowledge the significant contributions made to the quality of this product by Messrs. T...ILS ELEMENT E15COST ANALYSIS AND FUNDING DATA FLOW DIAGRAMS C .. i I, .. - I 4, - ., t .. •-! Ŗ 4 4 i -4- \\ o a _ __/ / ," I •i.\\ , Az!• "!-. * >Us

  3. High-Resolution Autoradiography

    DTIC Science & Technology

    1955-01-01

    Laboratory, Cleveland, Ohio WALTER C. WILLIAMS, B. S., Chief, High-Speed Flight Station, Edwards, Calif. HIItIU-ItE•,OL.I’TION Al’TIlT.AI) iIO (ltAIIII 3 Of )4r...comparison was made betw,,ia wvet-prociss autoraffio- eraluate this autoradiographic technique, several types of radio - graphs and autoradiographs...apart. heterogeneous system. The radiation emitted by the radio - Wet-process autoradiography, as developed in 1949 by Dr. active elements acts on a

  4. Application and Comparison of Stable Periodic Orbits in the Vicinity of Lagrangian Points L4 and L5 to a Four-Body Truth Model.

    DTIC Science & Technology

    1979-12-01

    IT/ /A/A79D-2 APPLICATION A-ND COMPARISON OF STABLE PERIODIC ORBITS IN THE VICXNyTy OF LAGRANGIAN POINTS L4 AND L5 TO A FOUR -BODY TRUTH MODEL THESISE...COMPARISON OF STABLE PERIODIC ORBITS IN TIlE ICINITY OF LAGRJ.NGIAN POINTS L4 AND LS TO A FOUR -BODY TRUTH MODEL. 11 ~ ’ THESIS PRESENTED TO THE FACULTY OF...15 Restricted Four -Body Equations of Motion .............. 18 III. Wheeler’s Orbit ......................................... 23 overview of Wheeler’s

  5. The PD1/PDL1 axis, a promising therapeutic target in aggressive breast cancers.

    PubMed

    Bertucci, François; Finetti, Pascal; Birnbaum, Daniel; Mamessier, Emilie

    2016-03-01

    Analysis of PDL1 mRNA expression in ∼5,500 breast cancers showed PDL1 upregulation in 38% of basal tumors and 38% of inflammatory breast cancers (IBC). Upregulation, associated with signs of strong cytotoxic local immune response, was associated with a better survival in the basal or triple-negative subtypes, and with a better pathological response to chemotherapy in these subtypes and IBC. Reactivation of dormant tumor-infiltrating lymphocytes (TILs) by PD1/PDL1-inhibitors represents a promising strategy in these aggressive tumors.

  6. Atomic and Molecular Physics Program

    DTIC Science & Technology

    2012-03-07

    i i å Sz i +1 -hzSz i -hxSx i hx =0: classical first order phase transition Finite hx: quantum phase transition, second order 13...transition H = J i å Sz iSz i +1 - (1- D) Sz i - 23/2 t Sx i D = E-U E: energy difference per lattice site, or lattice tilt U: onsite interaction...distribution is unlimited. Laser cooling Dysprosium \\.J ••• • ·=--·-Rtnes • Ce ., Til Periodic Table of the Elements [iO " I

  7. The PD1/PDL1 axis, a promising therapeutic target in aggressive breast cancers

    PubMed Central

    Bertucci, François; Finetti, Pascal; Birnbaum, Daniel; Mamessier, Emilie

    2016-01-01

    ABSTRACT Analysis of PDL1 mRNA expression in ∼5,500 breast cancers showed PDL1 upregulation in 38% of basal tumors and 38% of inflammatory breast cancers (IBC). Upregulation, associated with signs of strong cytotoxic local immune response, was associated with a better survival in the basal or triple-negative subtypes, and with a better pathological response to chemotherapy in these subtypes and IBC. Reactivation of dormant tumor-infiltrating lymphocytes (TILs) by PD1/PDL1-inhibitors represents a promising strategy in these aggressive tumors. PMID:27141340

  8. Manufacturing Methods and Technology for Digital Fault Isolation of Hybrid Microelectronic Assemblies.

    DTIC Science & Technology

    1982-03-01

    8217I I’ U F ~roef is 24 1lines by MI ) columns with video and inverse, vidleo d ispl ay mfodus. F~or the 111:1 protrun, 1(ijr ’-leirints of’ lIP) systemis...expansion, 2-15 ,%~~................. ............. . . . -. %% ." - .- Sect ion 2 - W ork< Aeooijshed Stibseet ionl I) - til *\\1t01 rohe Sy’.tomi...operating mode to automatic. • MI - Set AP operating mode to manual. * M2 - Set AP operating mode to test. • UP - Raise probe. e DN - Lower probe

  9. National Dam Safety Program. Watchung Lake Dam (NJ00767), Raritan River Basin, Stony Brook, Somerset County, New Jersey Phase I Inspection Report.

    DTIC Science & Technology

    1981-07-01

    ACCESSION NO. 3. RECIPIENT’S CATALOG NUMBER IDAEN/NAPi-53842/NJ00767-81/07 ~J.. ______________ 4. TIL E W-d ubtile)5. TYPE OF REPORT & PERIOD COVERED Phase...develop written operating procedures and a periodic maintenance plan to ensure the safety of Lhe dam within one year from the date of approval of this...written operating procedures and a periodic maintenance plan to ensure the safety of the dam within one year from the date of approval of this report

  10. Characterization of Anaerobic Chemical Processes in Reservoirs: Problem Description and Conceptual Model Formulation.

    DTIC Science & Technology

    1981-04-01

    CHARACTERIZATION OF ANAEROBIC CHEMICAL PROCESSES IN RESERVOIRS...ETC(U) APR al D GUNNISON, J M BRANNON UNCLASSIFIED WES/TR/E-8i-6 L 1111 1.01115 32...INISTRUCTIONS REPORT DOCUMENTATION PAGE RECINT. COMSPLETIG FORM 4. TIL adSb~)S. TYPE Of REPORT G PERIOD COySRED OIARACTERIZATION OFAAEROBIC QEMICAL...Engineers, U. S. Army Ap(J%8 Washington, D . C. 2031480r J 14. MONITORING AGENCY NAME & ADORESS(f different bass Conti.IIing Office) 1S. SECURITY CLASS

  11. Prognostic impact of the tumor-infiltrating regulatory T-cell (Foxp3+)/activated cytotoxic T lymphocyte (granzyme B+) ratio on resected left-sided pancreatic cancer

    PubMed Central

    Hwang, Ho Kyoung; Kim, Hyoung-Il; Kim, Se Hoon; Choi, Junjeong; Kang, Chang Moo; Kim, Kyung Sik; Lee, Woo Jung

    2016-01-01

    Among the subsets of tumor-infiltrating lymphocytes (TILs), activated cytotoxic T lymphocytes (granzyme B+) have an antitumor effect, while regulatory T lymphocytes [forkhead box P3 (Foxp3)+] suppress the antitumor immune response. The aim of the present study was to investigate the possible associations between TIL subsets and survival outcomes in patients with left-sided pancreatic ductal adenocarcinoma (PDAC). From January 2000 to December 2008, 30 patients who underwent curative distal pancreatectomy without neoadjuvant chemoradiotherapy due to left-sided PDAC were enrolled in the present study. TIL subsets were enumerated by immunohistochemical staining for cluster of differentiation (CD)3, CD4, CD8, Foxp3 and granzyme B in the intra-tumoral areas of tissue blocks. Patients were divided into two groups according to the median value of the absolute counts and relative ratios of TIL subsets. In the univariate analysis, age, gender, tumor size, nodal stage, tumor differentiation and lymphovascular/perineural invasion were not significantly associated with survival outcome. However, low levels of preoperative cancer antigen (CA) 19–9 were associated with a longer overall survival (OS), although the association was not significant (37 vs. 18 months; P=0.061). A high level of granzyme B+ was associated with enhanced disease-free survival (DFS) (25 vs. 10 months; P=0.023), and a low Foxp3+/granzyme B+ ratio was associated with a favorable prognosis in terms of DFS (25 vs. 8 months; P=0.008) and OS (47 vs. 17 months; P=0.003). In the multivariate analysis, the ratio of Foxp3+/granzyme B+ was an independent prognostic factor for determining DFS [Exp(B), 3.060; 95% confidence interval (CI), 1.259–47.436; P=0.014] and OS [Exp(B), 3.580; 95% CI, 1.460–8.780; P=0.005]. Among the clinicopathological factors, low levels of CA 19–9 were significantly associated with a low Foxp3+/granzyme B+ ratio (P=0.016). The results of the present study suggested that a low Foxp3

  12. On-the-Disk Development of the Halo Coronal Mass Ejection on 1998 May 2

    NASA Astrophysics Data System (ADS)

    Pohjolainen, S.; Maia, D.; Pick, M.; Vilmer, N.; Khan, J. I.; Otruba, W.; Warmuth, A.; Benz, A.; Alissandrakis, C.; Thompson, B. J.

    2001-07-01

    A halo coronal mass ejection (CME) was observed at 15:03 UT on 1998 May 2 by the Solar and Heliospheric Observatory Large-Angle Spectrometric Coronagraph. The observation of the CME was preceded by a major soft X-ray flare in NOAA Active Region 8210, characterized by a delta spot magnetic configuration and some activity in region 8214. A large transequatorial interconnecting loop (TIL) seen in the soft X-rays connected AR 8210 to a faint magnetic field region in the periphery of region 8214. Smaller loop systems were also connecting AR 8210 to other fainter bipolar magnetic structures, the interconnecting loop (IL) east of AR 8210 being one of the most visible. We present here a multiwavelength analysis of the large- and small-scale coronal structures associated with the development of the flare and of the CME, with emphasis placed on radio-imaging data. In the early phases of the flare, the radio emission sources traced the propagation paths of electrons along the TIL and the IL, which are accelerated in the vicinity of AR 8210. Furthermore, jetlike flows were observed in soft X-rays and in Hα in these directions. Significantly, the TIL and IL loop systems disappeared at least partially after the CME. An EUV Imaging Telescope (EIT) dimming region of similar size and shape to the soft X-ray TIL, but noticeably offset from it, was also observed. During the ``flash'' phase of the flare, new radio sources appeared, presenting signatures of destabilization and reconnection at discrete locations of the connecting loops. We interpret these as possible signatures of the CME liftoff on the disk. An Hα Moreton wave (blast wave) and an ``EIT wave'' were also observed, originating from the flaring AR 8210. The signatures in radio, after the wave propagated high into the corona, include type II-like emissions in the spectra. The radio images link these emissions to fast-moving sources, presumably formed at locations where the blast wave encounters magnetic structures. The

  13. First results of a search for double beta decay of {sup 100}Mo with the NEMO 2 detector

    SciTech Connect

    NEMO Collaboration

    1993-06-15

    Double beta decay of {sup 100}Mo (172g) is studied with the NEMO 2 detector in the Frejus Underground Laboratory. The experiment has now accumulated 2485 hours of data taking. A clear signal of 380 events for 2{beta}2{nu} decay has been obtained corresponding to a half-life of T{sub {1/2}} = 1.0 {plus_minus} 0.08 (syst.) 10{sup 19} y. Limits are presented for 2{beta}(0{nu}, {chi}), 2{beta}0{nu} (ground state and excited states 2{sub 1}{sup +} and 0{sub 1}{sup +}). The experiment will run til October 1993.

  14. Sleep problems and depression in adolescence: results from a large population-based study of Norwegian adolescents aged 16-18 years.

    PubMed

    Sivertsen, Børge; Harvey, Allison G; Lundervold, Astri J; Hysing, Mari

    2014-08-01

    Both sleep problems and depression are common problems in adolescence, but well-defined large epidemiological studies on the relationship are missing in this age group. The aim of this study was to examine the association between depression and several sleep parameters, including insomnia, in a population-based study of adolescents aged 16-18 years, and to explore potential gender differences. A large population-based study in Hordaland County in Norway conducted in 2012, the ung@hordaland study, surveyed 10,220 adolescents aged 16-18 years (54% girls) about sleep and depression. The sleep assessment included measures of the basic sleep parameters for weekdays and weekends. Depression was defined as scoring above the 90th percentile on the total score of Short Moods and Feelings Questionnaire (SMFQ). There was a large overlap between insomnia and depression in both genders and across depressive symptoms. Depressed adolescents exhibited significantly shorter sleep duration and time in bed as well as significantly longer sleep onset latency (SOL) and wake after sleep onset (WASO). Adolescents with insomnia had a 4- to 5-fold increased odds of depression compared to good sleepers. There was also a significant interaction between insomnia, sleep duration and depression, with a more than eightfold increase in odds of depression for those who met criteria for insomnia and who slept <6 h. These associations held for both genders, but were stronger in boys. To the best of our knowledge, this is the first population-based study to investigate sleep and insomnia in relation to depression among adolescents. The findings call for increased awareness of sleep problems and depression as a major public health issue.

  15. A Meta-Analysis: Identification of Common Mir-145 Target Genes that have Similar Behavior in Different GEO Datasets

    PubMed Central

    Ozgurses, Mete Emir; Demirel, Goksun; Aydin, Nizamettin; Ozen, Mustafa

    2016-01-01

    Background MicroRNAs, which are small regulatory RNAs, post-transcriptionally regulate gene expression by binding 3'-UTR of their mRNA targets. Their deregulation has been shown to cause increased proliferation, migration, invasion, and apoptosis. miR-145, an important tumor supressor microRNA, has shown to be downregulated in many cancer types and has crucial roles in tumor initiation, progression, metastasis, invasion, recurrence, and chemo-radioresistance. Our aim is to investigate potential common target genes of miR-145, and to help understanding the underlying molecular pathways of tumor pathogenesis in association with those common target genes. Methods Eight published microarray datasets, where targets of mir-145 were investigated in cell lines upon mir-145 over expression, were included into this study for meta-analysis. Inter group variabilities were assessed by box-plot analysis. Microarray datasets were analyzed using GEOquery package in Bioconducter 3.2 with R version 3.2.2 and two-way Hierarchical Clustering was used for gene expression data analysis. Results Meta-analysis of different GEO datasets showed that UNG, FUCA2, DERA, GMFB, TF, and SNX2 were commonly downregulated genes, whereas MYL9 and TAGLN were found to be commonly upregulated upon mir-145 over expression in prostate, breast, esophageal, bladder cancer, and head and neck squamous cell carcinoma. Biological process, molecular function, and pathway analysis of these potential targets of mir-145 through functional enrichments in PPI network demonstrated that those genes are significantly involved in telomere maintenance, DNA binding and repair mechanisms. Conclusion As a conclusion, our results indicated that mir-145, through targeting its common potential targets, may significantly contribute to tumor pathogenesis in distinct cancer types and might serve as an important target for cancer therapy. PMID:27655328

  16. Individual substitution mutations in the AID C terminus that ablate IgH class switch recombination.

    PubMed

    Kadungure, Tatenda; Ucher, Anna J; Linehan, Erin K; Schrader, Carol E; Stavnezer, Janet

    2015-01-01

    Activation-induced cytidine deaminase (AID) is essential for class switch recombination (CSR) and somatic hypermutation (SHM) of Ig genes. The C terminus of AID is required for CSR but not for SHM, but the reason for this is not entirely clear. By retroviral transduction of mutant AID proteins into aid-/- mouse splenic B cells, we show that 4 amino acids within the C terminus of mouse AID, when individually mutated to specific amino acids (R190K, A192K, L196S, F198S), reduce CSR about as much or more than deletion of the entire C terminal 10 amino acids. Similar to ΔAID, the substitutions reduce binding of UNG to Ig Sμ regions and some reduce binding of Msh2, both of which are important for introducing S region DNA breaks. Junctions between the IgH donor switch (S)μ and acceptor Sα regions from cells expressing ΔAID or the L196S mutant show increased microhomology compared to junctions in cells expressing wild-type AID, consistent with problems during CSR and the use of alternative end-joining, rather than non-homologous end-joining (NHEJ). Unlike deletion of the AID C terminus, 3 of the substitution mutants reduce DNA double-strand breaks (DSBs) detected within the Sμ region in splenic B cells undergoing CSR. Cells expressing these 3 substitution mutants also have greatly reduced mutations within unrearranged Sμ regions, and they decrease with time after activation. These results might be explained by increased error-free repair, but as the C terminus has been shown to be important for recruitment of NHEJ proteins, this appears unlikely. We hypothesize that Sμ DNA breaks in cells expressing these C terminus substitution mutants are poorly repaired, resulting in destruction of Sμ segments that are deaminated by these mutants. This could explain why these mutants cannot undergo CSR.

  17. Effects of Short-Term Thermal Alteration on Organic Matter in Experimentally-Heated Tagish Lake Observed by Raman Spectroscopy

    NASA Technical Reports Server (NTRS)

    Chan, Q. H. S.; Nakato, A.; Zolensky, M. E.; Nakamura, T.; Kebukawa, Y.; Maisano, J.; Colbert, M.; Martinez, J. E.

    2017-01-01

    Carbonaceous chondrites exhibit a wide range of aqueous and thermal alteration characteristics, while some are known to demonstrate mineralogical and petrologic evidence of having been thermally metamorphosed after aqueous alteration. This group of meteorites are commonly referred as thermally met-amorphosed carbonaceous chondrites (TMCCs), and their reflectance spectra show resemblances to that of C-type asteroids which typically have low albedos. This suggests that the surfaces of the C-type asteroids are also composed of both hydrous and dehydrated minerals, and thus TMCCs are among the best samples that can be studied in laboratory to reveal the true nature of the C-type asteroids. Although TMCCs are usually meteorites that were previously categorized as CI and CM chondrites, they are not strictly CI/CM because they exhibit isotopic and petrographic characteristics that significantly deviate from typical CI/CM. More appropriately, they are called CI-like and/or CM-like chondrites. Typical examples of TMCCs include the C2-ung/CM2TIV Belgica (B)-7904 and Yamato (Y) 86720. Thermal alteration is virtually complete in these meteorites and thus they are considered typical end-members of TMCCs exhibiting complete dehydration of matrix phyllosilicates. The estimated heating conditions are 10 to 103 days at 700 C to 1 to 100 hours at 890 C, i.e. short-term heating induced by impact and/or solar radiation. While the petrology and chemistry of TMCCs have only recently been extensively characterized, we have just begun to study in detail their organic contents. In order to understand how short-term heating affects the maturity of insoluble organic matter (IOM) in hydrous chondrites, we investigated experimentally-heated Tagish Lake meteorite using Raman spectroscopy, as the chemical and bulk oxygen isotopic compositions of the matrix of the carbonate (CO3)-poor lithology of the Tagish Lake (hereafter Tag) meteorite bears similarities to the TMCCs.

  18. NMR structure of the (1-51) N-terminal domain of the HIV-1 regulatory protein Vpr.

    PubMed

    Wecker, K; Roques, B P

    1999-12-01

    The human immunodeficiency virus type 1 (HIV-1) genome encodes a highly conserved 16 kDa regulatory gene product, Vpr (viral protein of regulation, 96 amino acid residues), which is incorporated into virions, in quantities equivalent to those of the viral Gag proteins. In the infected cells, Vpr is believed to function in the early phase of HIV-1 replication, including nuclear migration of preintegration complex, transcription of the provirus genome and viral multiplication by blocking cells in the G2 phase. Vpr has a critical role in long-term AIDS disease by inducing infection in nondividing cells such as monocytes and macrophages. Mutations have suggested that the N-terminal domain of Vpr encompassing the first 40 residues could be required for nuclear localization, packaging into virions and binding of transcription factor (TFIIB, Sp1), viral proteins (p6) and cellular proteins (RIP1, UNG, karyopherins). To gain insight into the structure-function relationship of Vpr, (1-51)Vpr was synthesized and its structure analyzed by circular dichroism and two-dimensional 1H NMR in aqueous trifluoroethanol (30%) solution and refined by restrained molecular dynamics. The structure is characterized by three turns around the first three prolines, Pro5, Pro10, Pro14, followed by a long amphipathic alpha helix-turn-alpha helix (Asp17-Ile46) motif ended by a turn extending from Tyr47 to Thr49. The alpha helix-turn-alpha helix motif and the amphipathic helix are well known for being implicated in protein-protein or protein-nucleic acid interaction. Therefore structural characteristics of the (1-51) N-terminal fragment of Vpr could explain why this region of Vpr plays a role in several biological functions of this protein.

  19. Cytosine-to-uracil deamination by SssI DNA methyltransferase.

    PubMed

    Stier, Ildikó; Kiss, Antal

    2013-01-01

    The prokaryotic DNA(cytosine-5)methyltransferase M.SssI shares the specificity of eukaryotic DNA methyltransferases (CG) and is an important model and experimental tool in the study of eukaryotic DNA methylation. Previously, M.SssI was shown to be able to catalyze deamination of the target cytosine to uracil if the methyl donor S-adenosyl-methionine (SAM) was missing from the reaction. To test whether this side-activity of the enzyme can be used to distinguish between unmethylated and C5-methylated cytosines in CG dinucleotides, we re-investigated, using a sensitive genetic reversion assay, the cytosine deaminase activity of M.SssI. Confirming previous results we showed that M.SssI can deaminate cytosine to uracil in a slow reaction in the absence of SAM and that the rate of this reaction can be increased by the SAM analogue 5'-amino-5'-deoxyadenosine. We could not detect M.SssI-catalyzed deamination of C5-methylcytosine ((m5)C). We found conditions where the rate of M.SssI mediated C-to-U deamination was at least 100-fold higher than the rate of (m5)C-to-T conversion. Although this difference in reactivities suggests that the enzyme could be used to identify C5-methylated cytosines in the epigenetically important CG dinucleotides, the rate of M.SssI mediated cytosine deamination is too low to become an enzymatic alternative to the bisulfite reaction. Amino acid replacements in the presumed SAM binding pocket of M.SssI (F17S and G19D) resulted in greatly reduced methyltransferase activity. The G19D variant showed cytosine deaminase activity in E. coli, at physiological SAM concentrations. Interestingly, the C-to-U deaminase activity was also detectable in an E. coli ung (+) host proficient in uracil excision repair.

  20. Datenkompression, Prozessoptimierung, Aerodynamik: Eine Tour Durch die Skalen

    NASA Astrophysics Data System (ADS)

    Dahmen, Wolfgang; Marquardt, Wolfgang

    Enorm gestiegene Rechnerkapazitäten ermutigen zu einer immer genaueren Erschließung komplexer Prozesse in Natur- und Ingenieurwissenschaften durch numerische Simulation. Die Mathematik als Schnittstelle zwischen realer und digitaler Welt liefert einerseits die Grundlagen zur Formulierung notwendigerweise vereinfachter Modelle der Realität. Andererseits bietet sie aber auch die methodischen Grundlagen für den Entwurf effizienter Algorithmen, die aus solchen Modellen quantitative Information gewinnen können. Eine zentrale Herausforderung liegt dabei in der Tatsache, dass reale Prozesse meist durch relevante Anteile auf einer riesigen Spannweite von Längenskalen getrieben werden. Die Entwicklung mathematischer Methoden zur Behandlung von Mehrskaligkeit ist daher von wesentlicher Bedeutung. In diesem Artikel wird dies anhand jüngster Entwicklungen in ganz unterschiedlichen Anwendungsbereichen erklärt und illustriert, auch um deutlich zu machen, wieviele "Fliegen“ man mit einer "mathematischen Klappe“ schlagen kann. Insbesondere werden grundlegende, auf Waveletzerlegung beruhende Prinzipien zunächst im Rahmen der Bildkompression und -kodierung auf weitgehend elementare Weise erklärt. Es wird dann aufgezeigt, dass diese Konzepte in ganz ähnlicher Weise bei der Datenanalyse im Zusammenhang mit verfahrenstechnischen Anwendungen, bei großskaligen Optimierungsproblemen der Prozessindustrie sowie bei komplexen Strömungsproblemen wie in aerodynamischen Anwendungen zum Tragen kommen. Stabile Zerlegungen in Anteile unterschiedlicher Längenskalen eröffnen dabei einen Zugang zu adaptiven Lösungskonzepten, die in der Lage sind, automatisch Rechnerresourcen dort zu plazieren, wo sie zu einer ökonomischen Realisierung der gewünschten Lösungsqualität, etwa in Form von Genauigkeitstoleranzen, benötigt werden.

  1. Changes in the distribution of the grey mangrove Avicennia marina (Forsk.) using large scale aerial color infrared photographs: are the changes related to habitat modification for mosquito control?

    NASA Astrophysics Data System (ADS)

    Jones, J.; Dale, P. E. R.; Chandica, A. L.; Breitfuss, M. J.

    2004-09-01

    Runnelling, a method of habitat modification used for mosquito management in intertidal saltmarshes in Australia, alters marsh hydrology. The objective of this research was to assess if runnelling had affected the distribution of the grey mangrove ( Avicennia marina (Forsk.)) at a study site in southeast Queensland. Since runnelling is carried out in diverse marshes a second aim was to assess differences in mangrove colonisation in the two main saltmarsh species in the area. These are marine couch [ Sporobolus virginicus (L.) Kunth.] and samphire [ Sarcocornia quinqueflora (Bunge ex Ung.-Stern.)]. Runnels at the study site were in an area dominated by Sporobolus. The mangrove area was measured by classifying digital color infrared (CIR) data obtained from aerial photographs acquired in 1982, which was 3 years before runnelling, and in 1987, 1991 and 1999, 2-14 years after. Changes in the spatial extent of A. marina were identified using difference images produced from post-classification change detection. The results showed that runnels did not significantly influence the distribution of A. marina at the study site. At a more detailed level differences in A. marina establishment in the Sporobolus and Sarcocornia areas were determined from counts of trees on the aerial photographs. There was a greater proportion of mangroves in Sarcocornia than Sporobolus and this increased over time. This may be related to differences in density between the plant species, to grapsid crab activity or to other edaphic conditions. There may be implications for runnelling in Sarcocornia marshes. The large increase observed in A. marina in the area generally is likely to be related to factors such as catchment modification or tidal/sea-level changes. It is concluded that runnelling has not led to mangrove establishment in the Sporobolus dominated saltmarsh.

  2. Altered Gene Expressions and Cytogenetic Repair Efficiency in Cells with Suppressed Expression of XPA after Proton Exposure

    NASA Technical Reports Server (NTRS)

    Zhang, Ye; Rohde, Larry H.; Gridley, Daila S.; Mehta, Satish K.; Pierson, Duane L.; Wu, Honglu

    2009-01-01

    Cellular responses to damages from ionizing radiation (IR) exposure are influenced not only by the genes involved in DNA double strand break (DSB) repair, but also by non- DSB repair genes. We demonstrated previously that suppressed expression of several non-DSB repair genes, such as XPA, elevated IR-induced cytogenetic damages. In the present study, we exposed human fibroblasts that were treated with control or XPA targeting siRNA to 250 MeV protons (0 to 4 Gy), and analyzed chromosome aberrations and expressions of genes involved in DNA repair. As expected, after proton irradiation, cells with suppressed expression of XPA showed a significantly elevated frequency of chromosome aberrations compared with control siRNA treated (CS) cells. Protons caused more severe DNA damages in XPA knock-down cells, as 36% cells contained multiple aberrations compared to 25% in CS cells after 4Gy proton irradiation. Comparison of gene expressions using the real-time PCR array technique revealed that expressions of p53 and its regulated genes in irradiated XPA suppressed cells were altered similarly as in CS cells, suggesting that the impairment of IR induced DNA repair in XPA suppressed cells is p53-independent. Except for XPA, which was more than 2 fold down regulated in XPA suppressed cells, several other DNA damage sensing and repair genes (GTSE1, RBBP8, RAD51, UNG and XRCC2) were shown a more than 1.5 fold difference between XPA knock-down cells and CS cells after proton exposure. The possible involvement of these genes in the impairment of DNA repair in XPA suppressed cells will be further investigated.

  3. Habitat characteristics and eggshell distribution of the salt marsh mosquito, Aedes vigilax, in marshes in subtropical Eastern Australia.

    PubMed

    Dale, Pat E R; Knight, Jon; Kay, Brian H; Chapman, Heather; Ritchie, Scott A; Brown, Michael D

    2008-01-01

    Research at 10 locations in coastal subtropical Queensland, Australia, has shown that salt marshes contained heterogeneous distributions of eggshells of the pest and vector mosquito Aedes vigilax (Skuse) (Diptera:Culicidae). The eggshell distribution was related to specific vegetation assemblages, with a mix of the grass, Sporobolus virginicus (L.) Kunth (Poales: Poaceae), and the beaded glasswort, Sarcocornia quinqueflora (Bunge ex (Ung.-Stern) A.J. Scott (Caryophyllales: Chenopodiaceae), as significantly higher in eggshells than any other vegetation. There were also high numbers in the mix of S. virginicus with the arrowgrass, Triglochin striata Ruiz & Pavón (Alismatales: Juncaginaceae). Both mixed types are found in relatively wetter areas, despite very few eggshells being found generally in the low marsh. Most sites contained S. virginicus and eggshell locations were variable for this species alone. This was probably related to its life form variability in response to salinity and location on the marsh. Location on the marsh was important for eggshell distribution with most eggshells around the edges of pools and depressions, followed by, but to a significantly lesser extent, the marsh surface. Eggshells were fewest in the low marsh. Partition analysis resulted in a tree that simplified and summarised the factors important for eggshell distribution confirming the individual analyses. The potential effects of climate, sea level and other change are also briefly discussed in the context of likely changes to land cover and relative location on the marsh. For example, increased sea level may lead to low marsh conditions extending into higher marsh area with implications for oviposition and numbers of eggshells.

  4. Base excision repair activities in organotypic hippocampal slice cultures exposed to oxygen and glucose deprivation.

    PubMed

    Rolseth, Veslemøy; Rundén-Pran, Elise; Neurauter, Christine Gran; Yndestad, Arne; Luna, Luisa; Aukrust, Pål; Ottersen, Ole Petter; Bjørås, Magnar

    2008-06-01

    The capacity for DNA repair is likely to be one of the factors that determine the vulnerability of neurons to ischemic stress and may influence the pathological outcome of stroke. In this report, initiation of base excision repair (BER) was assessed by analysis of enzyme activity and gene expression level of DNA glycosylases and AP-endonucleases in rat organotypic hippocampal slice cultures exposed to oxygen and glucose deprivation (OGD) - an in vitro model of stroke. Under basal conditions, AP-endonuclease activity and base removal of ethenoadenine and 8-oxoguanine (8-oxoG) were higher (by approximately 20-35 %) in CA3/fascia dentata (FD) than in CA1. Base removal of uracil did not differ between the two hippocampal regions, while removal of 5-hydroxyuracil (5-OHU) was slightly less efficient in CA3/FD than in CA1. Analyses performed immediately after 30 min of OGD revealed a decreased AP-endonuclease activity (by approximately 20%) in CA1 as well as CA3/FD, and an increased ethenoadenine activity (by approximately 25%) in CA1. Activities for 8-oxoG, 5-OHU and uracil showed no significant changes at this time point. At 8h after OGD, none of the enzyme activities differed from control values. Real-time RT-PCR showed that transcription of DNA glycosylases, including Ogg1, Nth1, Ung, Aag, Neil1 and Neil2 were not changed in response to OGD treatment (t=0 h). The hippocampal expression of Neil2 was low compared with the other DNA glycosylases. These data indicate that CA1 has a lower capacity than CA3/FD for removal of base lesions under basal conditions. The relatively low capacity for BER in basal conditions and the apparent failure to upregulate repair of oxidative damage after OGD might contribute to the high vulnerability of CA1 to ischemic injury.

  5. Non-small cell lung cancer exhibits transcript overexpression of genes associated with homologous recombination and DNA replication pathways.

    PubMed

    Saviozzi, Silvia; Ceppi, Paolo; Novello, Silvia; Ghio, Paolo; Lo Iacono, Marco; Borasio, Piero; Cambieri, Alberto; Volante, Marco; Papotti, Mauro; Calogero, Raffaele A; Scagliotti, Giorgio V

    2009-04-15

    Genes involved in DNA repair and replication have been recently investigated as predictive markers of response to chemotherapy in non-small cell lung cancer (NSCLC). However, few data on the expression of these genes in tumor compared with corresponding normal lung are available. The aim of this study was to evaluate differential mRNA levels of 22 DNA repair genes of five different DNA repair pathways: direct, base excision, nucleotide excision (NER), double-strand break (DSBR), and postreplicative repair. In addition, six genes involved in DNA replication (REP) and three telomere maintenance genes were investigated. Total RNAs extracted from fresh-frozen tumors and corresponding normal tissues of 50 consecutive chemo-naïve resected NSCLC patients were analyzed. Transcript levels were quantified by real-time PCR. A significant overexpression was detected in 20 of 30 (67%) genes, mostly belonging to DSBR pathways, whereas others (XPA, XPC, and UBE2N; 10%) were significantly underexpressed. For 7 of 30 (23%) genes, mostly belonging to NER pathway, no significant difference between paired tumor and normal samples was observed. Transcript overexpression of DSBR and REP genes was significantly higher in poorly differentiated carcinomas and DSBR levels were higher in men compared with women. The transcriptional overexpression of four genes (XRCC5, TOP3B, TYMS, and UNG) showed significant correlation with a shorter patients' outcome at the univariate, whereas only stage of disease appeared as an independent factor affecting prognosis, as assessed by multivariate analysis. In conclusion, genes belonging to DNA repair/replication pathways are overexpressed in NSCLC and are associated with a more aggressive phenotype.

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

    PubMed

    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.

  7. Co-expression of TIM-3 and CEACAM1 promotes T cell exhaustion in colorectal cancer patients.

    PubMed

    Zhang, Yang; Cai, Pengcheng; Li, Lei; Shi, Liang; Chang, Panpan; Liang, Tao; Yang, Qianqian; Liu, Yang; Wang, Lin; Hu, Lihua

    2017-02-01

    T-cell immunoglobulin domain and mucin domain-3(TIM-3) is an activation induced inhibitory molecule involved in immune tolerance and is recently reported to induce T cell exhaustion which is mediated by carcinoembryonic antigen cell adhesion molecule 1(CEACAM1), another well-known molecule expressed on activated T cells and involved in T cell inhibition. To investigate the expression of TIM-3 and CEACAM1 on circulating CD8(+) T cells and tumor infiltrating lymphocytes (TILs), 65 diagnosed colorectal cancer (CRC) patients and 38 healthy controls were enrolled in this study and the results showed that TIM-3 and CEACAM1 were both highly expressed on circulating CD8(+) T cells in CRC patients and elevated on TILs compared with paraneoplastic T cells. Furthermore, TIM-3(+)CEACAM1(+) CD8(+) T cells represented the most dysfunctional population with the least IFN-γ production. In addition, the expressions of TIM-3 and CEACAM1 were correlated with advanced stage and could be independent risk factors for CRC. We for the first time to our knowledge suggested that co-expression of TIM-3 and CEACAM1 can mediate T cell exhaustion and may be potential biomarkers for CRC prediction, highlighting the possibility of being immunotherapy targets.

  8. Use of HLA peptidomics and whole exome sequencing to identify human immunogenic neo-antigens

    PubMed Central

    Kalaora, Shelly; Qutob, Nouar; Teer, Jamie K.; Shimony, Nilly; Schachter, Jacob; Rosenberg, Steven A.; Samuels, Yardena

    2016-01-01

    The antigenicity of cells is demarcated by the peptides bound by their Human Leucocyte Antigen (HLA) molecules. Through this antigen presentation, T cell specificity response is controlled. As a fraction of the expressed mutated peptides is presented on the HLA, these neo-epitopes could be immunogenic. Such neoantigens have recently been identified through screening for predicted mutated peptides, using synthetic peptides or ones expressed from minigenes, combined with screening of patient tumor-infiltrating lymphocytes (TILs). Here we present a time and cost-effective method that combines whole-exome sequencing analysis with HLA peptidome mass spectrometry, to identify neo-antigens in a melanoma patient. Of the 1,019 amino acid changes identified through exome sequencing, two were confirmed by mass spectrometry to be presented by the cells. We then synthesized peptides and evaluated the two mutated neo-antigens for reactivity with autologous bulk TILs, and found that one yielded mutant-specific T-cell response. Our results demonstrate that this method can be used for immune response prediction and promise to provide an alternative approach for identifying immunogenic neo-epitopes in cancer. PMID:26819371

  9. Isolation of neoantigen-specific T cells from tumor and peripheral lymphocytes

    PubMed Central

    Cohen, Cyrille J.; Gartner, Jared J.; Horovitz-Fried, Miryam; Shamalov, Katerina; Trebska-McGowan, Kasia; Bliskovsky, Valery V.; Parkhurst, Maria R.; Ankri, Chen; Prickett, Todd. D.; Crystal, Jessica S.; Li, Yong F.; El-Gamil, Mona; Rosenberg, Steven A.; Robbins, Paul F.

    2015-01-01

    Adoptively transferred tumor-infiltrating T lymphocytes (TILs) that mediate complete regression of metastatic melanoma have been shown to recognize mutated epitopes expressed by autologous tumors. Here, in an attempt to develop a strategy for facilitating the isolation, expansion, and study of mutated antigen–specific T cells, we performed whole-exome sequencing on matched tumor and normal DNA isolated from 8 patients with metastatic melanoma. Candidate mutated epitopes were identified using a peptide-MHC–binding algorithm, and these epitopes were synthesized and used to generate panels of MHC tetramers that were evaluated for binding to tumor digests and cultured TILs used for the treatment of patients. This strategy resulted in the identification of 9 mutated epitopes from 5 of the 8 patients tested. Cells reactive with 8 of the 9 epitopes could be isolated from autologous peripheral blood, where they were detected at frequencies that were estimated to range between 0.4% and 0.002%. To the best of our knowledge, this represents the first demonstration of the successful isolation of mutation-reactive T cells from patients’ peripheral blood prior to immune therapy, potentially providing the basis for designing personalized immunotherapies to treat patients with advanced cancer. PMID:26389673

  10. Long-term prospects for the environmental profile of advanced sugar cane ethanol.

    PubMed

    da Silva, Cinthia R U; Franco, Henrique Coutinho Junqueira; Junqueira, Tassia Lopes; van Oers, Lauran; van der Voet, Ester; Seabra, Joaquim E A

    2014-10-21

    This work assessed the environmental impacts of the production and use of 1 MJ of hydrous ethanol (E100) in Brazil in prospective scenarios (2020-2030), considering the deployment of technologies currently under development and better agricultural practices. The life cycle assessment technique was employed using the CML method for the life cycle impact assessment and the Monte Carlo method for the uncertainty analysis. Abiotic depletion, global warming, human toxicity, ecotoxicity, photochemical oxidation, acidification, and eutrophication were the environmental impacts categories analyzed. Results indicate that the proposed improvements (especially no-til farming-scenarios s2 and s4) would lead to environmental benefits in prospective scenarios compared to the current ethanol production (scenario s0). Combined first and second generation ethanol production (scenarios s3 and s4) would require less agricultural land but would not perform better than the projected first generation ethanol, although the uncertainties are relatively high. The best use of 1 ha of sugar cane was also assessed, considering the displacement of the conventional products by ethanol and electricity. No-til practices combined with the production of first generation ethanol and electricity (scenario s2) would lead to the largest mitigation effects for global warming and abiotic depletion. For the remaining categories, emissions would not be mitigated with the utilization of the sugar cane products. However, this conclusion is sensitive to the displaced electricity sources.

  11. A Chimeric Switch-Receptor Targeting PD1 Augments the Efficacy of Second-Generation CAR T Cells in Advanced Solid Tumors.

    PubMed

    Liu, Xiaojun; Ranganathan, Raghuveer; Jiang, Shuguang; Fang, Chongyun; Sun, Jing; Kim, Soyeon; Newick, Kheng; Lo, Albert; June, Carl H; Zhao, Yangbing; Moon, Edmund K

    2016-03-15

    Chimeric antigen receptor (CAR)-modified adoptive T-cell therapy has been successfully applied to the treatment of hematologic malignancies, but faces many challenges in solid tumors. One major obstacle is the immune-suppressive effects induced in both naturally occurring and genetically modified tumor-infiltrating lymphocytes (TIL) by inhibitory receptors (IR), namely PD1. We hypothesized that interfering with PD1 signaling would augment CAR T-cell activity against solid tumors. To address this possibility, we introduced a genetically engineered switch receptor construct, comprising the truncated extracellular domain of PD1 and the transmembrane and cytoplasmic signaling domains of CD28, into CAR T cells. We tested the effect of this supplement, "PD1CD28," on human CAR T cells targeting aggressive models of human solid tumors expressing relevant tumor antigens. Treatment of mice bearing large, established solid tumors with PD1CD28 CAR T cells led to significant regression in tumor volume due to enhanced CAR TIL infiltrate, decreased susceptibility to tumor-induced hypofunction, and attenuation of IR expression compared with treatments with CAR T cells alone or PD1 antibodies. Taken together, our findings suggest that the application of PD1CD28 to boost CAR T-cell activity is efficacious against solid tumors via a variety of mechanisms, prompting clinical investigation of this potentially promising treatment modality.

  12. HAGE (DDX43) is a biomarker for poor prognosis and a predictor of chemotherapy response in breast cancer

    PubMed Central

    Abdel-Fatah, T M A; McArdle, S E B; Johnson, C; Moseley, P M; Ball, G R; Pockley, A G; Ellis, I O; Rees, R C; Chan, S Y T

    2014-01-01

    Background: HAGE protein is a known immunogenic cancer-specific antigen. Methods: The biological, prognostic and predictive values of HAGE expression was studied using immunohistochemistry in three cohorts of patients with BC (n=2147): early primary (EP-BC; n=1676); primary oestrogen receptor-negative (PER-BC; n=275) treated with adjuvant anthracycline-combination therapies (Adjuvant-ACT); and primary locally advanced disease (PLA-BC) who received neo-adjuvant anthracycline-combination therapies (Neo-adjuvant-ACT; n=196). The relationship between HAGE expression and the tumour-infiltrating lymphocytes (TILs) in matched prechemotherapy and postchemotherapy samples were investigated. Results: Eight percent of patients with EP-BC exhibited high HAGE expression (HAGE+) and was associated with aggressive clinico-pathological features (Ps<0.01). Furthermore, HAGE+expression was associated with poor prognosis in both univariate and multivariate analysis (Ps<0.001). Patients with HAGE+did not benefit from hormonal therapy in high-risk ER-positive disease. HAGE+and TILs were found to be independent predictors for pathological complete response to neoadjuvant-ACT; P<0.001. A statistically significant loss of HAGE expression following neoadjuvant-ACT was found (P=0.000001), and progression-free survival was worse in those patients who had HAGE+residual disease (P=0.0003). Conclusions: This is the first report to show HAGE to be a potential prognostic marker and a predictor of response to ACT in patients with BC. PMID:24755885

  13. Temperature-controlled ionic liquid-based ultrasound-assisted microextraction for preconcentration of trace quantity of cadmium and nickel by using organic ligand in artificial saliva extract of smokeless tobacco products

    NASA Astrophysics Data System (ADS)

    Arain, Sadaf Sadia; Kazi, Tasneem Gul; Arain, Asma Jabeen; Afridi, Hassan Imran; Baig, Jameel Ahmed; Brahman, Kapil Dev; Naeemullah; Arain, Salma Aslam

    2015-03-01

    A new approach was developed for the preconcentration of cadmium (Cd) and nickel (Ni) in artificial saliva extract of dry snuff (brown and black) products using temperature-controlled ionic liquid-based ultrasound-assisted dispersive liquid-liquid microextraction (TIL-UDLLμE) followed by electrothermal atomic absorption spectrometry (ETAAS). The Cd and Ni were complexed with ammonium pyrrolidinedithiocarbamate (APDC), extracted in ionic liquid drops, 1-butyl-3-methylimidazolium hexafluorophosphate [C4MIM][PF6]. The multivariate strategy was applied to estimate the optimum values of experimental variables influence the % recovery of analytes by TIL-UDLLμE method. At optimum experimental conditions, the limit of detection (3s) were 0.05 and 0.14 μg L-1 while relative standard deviations (% RSD) were 3.97 and 3.55 for Cd and Ni respectively. After extraction, the enhancement factors (EF) were 87 and 79 for Cd and Ni, respectively. The RSD for six replicates of 10 μg L-1 Cd and Ni were 3.97% and 3.55% respectively. To validate the proposed method, certified reference material (CRM) of Virginia tobacco leaves was analyzed, and the determined values of Cd and Ni were in good agreement with the certified values. The concentration of Cd and Ni in artificial saliva extracts corresponds to 39-52% and 21-32%, respectively, of the total contents of both elements in dry brown and black snuff products.

  14. Anatomical study of the posterior cruciate ligament with the knee flexed at 90°☆☆☆

    PubMed Central

    Cho, Daniel Kyubin; Rosa, Sthéphano Pellizzaro; Prestes, Guilherme Bello; da Cunha, Luiz Antônio Munhoz; de Moura, Márcio Fernando Aparecido; Stieven Filho, Edmar

    2014-01-01

    Objective To study the anatomy of the posterior cruciate ligament (PCL) and define anatomical parameters with the knee flexed at 90°. Methods Eight knees from cadavers were dissected in order to make measurements from the center of the anterolateral band to the roof (AL1), from the center of the anterolateral band to the anterior cartilage (AL2), from the center of the posteromedial band to the roof (PM1), from the center of the posteromedial band to the anterior cartilage (PM2), from the center of the tibial insertion to the medial region of the tibia (TIM), from the center of the tibial insertion to the lateral region of the tibia (TIL), from the center of the medial insertion to the medial meniscus (IMM) and the width of the origin of the PCL (WO). To obtain the results from each anatomical structure, the means and standard deviations of the measurements were calculated. Results The measurements in millimeters that were found were AL1, 6.2; AL2, 4.9; PM1, 11.7; PM2, 5.5; TIM, 32.5; TIL, 40.6; IMM, 9.4; and WO, 32.5. Conclusions The PCL has an extensive origin. The center of the anterolateral band is 6 mm from the roof and 5 mm from the anterior cartilage of the knee. The tibial insertion is slightly medial and 10 mm distal to the posterior cornu of the medial meniscus. PMID:26229851

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

    PubMed

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

    2015-05-01

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

  16. BmSI-7, a novel subtilisin inhibitor from Boophilus microplus, with activity toward Pr1 proteases from the fungus Metarhizium anisopliae.

    PubMed

    Sasaki, Sergio D; de Lima, Cássia A; Lovato, Diogo V; Juliano, Maria A; Torquato, Ricardo J S; Tanaka, Aparecida S

    2008-02-01

    BmSI-7 and BmSI-6, two Boophilus microplus subtilisin inhibitors (BmSI) were purified and characterized from eggs. The inhibitors isolated by classical purification methods presented molecular masses of 7408 and 7271Da, respectively, by MALDI-TOF-MS. Both BmSI-7 and BmSI-6 inhibited neutrophil elastase (K(i) 0.4 and 0.3nM) and subtilisin A (K(i) 1.4nM for both inhibitors). They also strongly inhibited Pr1 proteases from the fungus Metarhizium anisopliae; BmSI-7 (K(i) 50nM) and BmSI-6 (K(i) 2.2nM). The BmSI-7 full length cDNA was obtained using amino acid sequence information of BmSI-7 peptides generated by proteolytic digestion. BmSI-7 belongs to trypsin inhibitor like cysteine rich domain family (TIL), and it is transcribed in ovary, fat body, gut, salivary gland and haemocytes. BmSI-7 is the first TIL inhibitor described with inhibitory activity toward subtilisin A and Pr1 proteases of entomopathogenic fungi.

  17. Characteristics of nanocomposite ZrO2/Al2O3 films deposited by plasma-enhanced atomic layer deposition.

    PubMed

    Yun, Sun Jin; Lim, Jung Wook; Kim, Hyun-Tak

    2007-11-01

    Nanocomposite ZrO2/Al2O3 (ZAO) films were deposited on Si by plasma-enhanced atomic layer deposition and the film characteristics including interfacial oxide formation, dielectric constant (k), and electrical breakdown strength were investigated without post-annealing process. In both the mixed and nano-laminated ZAO films, the thickness of the interfacial oxide layer (T(IL)) was considerably reduced compared to ZrO2 and Al2O3 films. The T(IL) was 0.8 nm in nano-composite films prepared at a mixing ratio (ZrO2:Al2O3) of 1:1. The breakdown strength and the leakage current level were greatly improved by adding Al2O3 as little as 7.9% compared to that of ZrO2 and were enhanced more with increasing content of Al2O3. The k of ZrO2 and mixed ZAO (Al2O3 7.9%) films were 20.0 and 16.5, respectively. These results indicate that the addition of Al2O3 to ZrO2 greatly improves the electrical properties with less cost of k compared to the addition of SiO2.

  18. The effect of preoperative local interleukin-2 (IL-2) injections in patients with head and neck squamous cell carcinoma. An immunological study.

    PubMed

    Cortesina, G; De Stefani, A; Galeazzi, E; Bussi, M; Giordano, C; Cavallo, G P; Jemma, C; Vai, S; Forni, G; Valente, G

    1991-01-01

    Clinical, immunological, immunophenotypical, pathological and molecular biological studies were performed on tumor infiltrating lymphocytes (TIL) and lymph node lymphocytes (LN-ly) of 8 patients with squamous cell carcinoma of the oral cavity and oropharynx treated with 10 daily locoregional injections of low doses of IL-2 before surgery. No complications were seen during or after surgery. In 3 cases the LN-ly showed a moderate LAK activity, higher in the LN-ly omolateral to the tumor and near the IL-2 injection site; in 2 of these 3 patients a good LAK activity was induced after 6-day culture with IL-2. The LN-ly derived from nodes next to the tumor showed a decreased NK activity and proliferative ability both in basal conditions and after in vitro lymphokine challenging. LN-ly of 2 IL-2 treated patients showed high levels of mRNA encoding for IL-2-R, while it was absent in 2 untreated cases. Immunophenotypical studies on TIL showed statistically improved levels of CD25+ and LAK1+ cells in treated cases. Clusters of CD11c+ (macrophages) cells were seen close to the neoplastic sheets.

  19. Comparison of circulating and intratumoral regulatory T cells in patients with renal cell carcinoma.

    PubMed

    Asma, Gati; Amal, Gorrab; Raja, Marrakchi; Amine, Derouiche; Mohammed, Chebil; Amel, Ben Ammar Elgaaied

    2015-05-01

    The clear evidence that tumor-infiltrating lymphocytes (TIL) exists in the tumor microenvironment raises the question why renal cell carcinoma (RCC) progresses. Numerous studies support the implication of CD4(+)CD25(high) regulatory T (Treg) cells in RCC development. We aimed in this study to characterize the phenotype and function of circulating and intratumoral Treg cells of RCC patient in order to evaluate their implication in the inhibition of the local antitumor immune response. Our results demonstrate that the proportion of Treg in TIL was, in average, similar to that found in circulating CD4(+) T cells of patients or healthy donors. However, intratumoral Treg exhibit a marked different phenotype when compared with the autologous circulating Treg. A higher CD25 mean level, HLA-DR, Fas, and GITR, and a lower CD45RA expression were observed in intratumoral Treg, suggesting therefore that these cells are effector in the tumor microenvironment. Additionally, intratumoral Treg showed a higher inhibitory function on autologous CD4(+)CD25(-) T cells when compared with circulating Treg that may be explained by an overexpression of FoxP3 transcription factor. These findings suggest that intratumoral Treg could be major actors in the impairment of local antitumor immune response for RCC patients.

  20. DPP4 truncated GM-CSF & IL-3 manifest distinct receptor binding & regulatory functions compared to their full length forms.

    PubMed

    O'Leary, H A; Capitano, M; Cooper, S; Mantel, C; Boswell, H S; Kapur, R; Ramdas, B; Chan, R; Deng, L; Qu, C-K; Broxmeyer, H E

    2017-03-27

    Dipeptidylpeptidase 4 (DPP4/CD26) enzymatically cleaves select penultimate amino acids of proteins, including colony stimulating factors (CSFs), and has been implicated in cellular regulation. To better understand the role of DPP4 regulation of hematopoiesis, we analyzed the activity of DPP4 on the surface of immature blood cells and then comparatively assessed the interactions and functional effects of full-length (FL) and DPP4 truncated factors [(T)-GM-CSF and- IL-3] on both in vitro and in vivo models of normal and leukemic cells. T-GM-CSF and T-IL-3 had enhanced receptor binding, but decreased CSF activity, compared to their FL forms. Importantly, T-GM-CSF and T-IL-3 significantly, and reciprocally, blunted receptor binding and myeloid progenitor cell proliferation activity of both FL-GM-CSF and FL-IL-3 in vitro and in vivo. Similar effects were apparent in vitro using cluster forming cells from patients with Acute Myeloid Leukemia (AML) regardless of cytogenetic or molecular alterations and in vivo utilizing animal models of leukemia. This suggests that DPP4 T-molecules have modified binding and functions compared to their FL counterparts and may serve regulatory roles in normal and malignant hematopoiesis.Leukemia accepted article preview online, 27 March 2017. doi:10.1038/leu.2017.98.

  1. Correlation between anti-PD-L1 tumor concentrations and tumor-specific and nonspecific biomarkers in a melanoma mouse model

    PubMed Central

    Contreras, Ana M.; Merino, María; Vasquez, Marcos; Trocóniz, Iñaki F.

    2016-01-01

    Blockade of PD-L1 with specific monoclonal antibodies (anti-PD-L1) represents a therapeutic strategy to increase the capability of the immune system to modulate the tumor immune-resistance. The relationship between anti-PD-L1 tumor exposition and anti-tumor effect represents a challenge that has been addressed in this work through the identification of certain biomarkers implicated in the antibody's mechanism of action, using a syngeneic melanoma mouse model. The development of an in-vitro/in-vivo platform has allowed us to investigate the PD-L1 behavior after its blockage with anti-PD-L1 at cellular level and in animals. In-vitro studies showed that the complex PD-L1/anti-PD-L1 was retained mainly at the cell surface. The antibody concentration and time exposure affected directly the recycling or ligand turnover. In-vivo studies showed that anti-PD-L1 was therapeutically active at all stage of the disease, with a rapid onset, a low but durable efficacy and non-relevant toxic effect. This efficacy measured as tumor shrinkage correlated with tumor-specific infiltrating lymphocytes (TILs), which increased as antibody tumor concentrations increased. Both, TILS and antibody concentrations followed similar kinetic patterns, justifying the observed anti-PD-L1 rapid onset. Interestingly, peripheral lymphocytes (PBLs) behave as infiltrating lymphocytes, suggesting that these PBLs might be considered as a possible biomarker for antibody activity. PMID:27764774

  2. Barrel time-of-flight detector for the PANDA experiment at FAIR

    NASA Astrophysics Data System (ADS)

    Gruber, L.; Brunner, S. E.; Marton, J.; Orth, H.; Suzuki, K.

    2016-07-01

    The barrel time-of-flight detector for the PANDA experiment at FAIR is foreseen as a Scintillator Tile (SciTil) Hodoscope based on several thousand small plastic scintillator tiles read-out with directly attached Silicon Photomultipliers (SiPMs). The main tasks of the system are an accurate determination of the time origin of particle tracks to avoid event mixing at high collision rates, relative time-of-flight measurements as well as particle identification in the low momentum regime. The main requirements are the use of a minimum material amount and a time resolution of σ < 100 ps. We have performed extensive optimization studies and prototype tests to prove the feasibility of the SciTil design and finalize the R&D phase. In a 2.7 GeV/c proton beam at Forschungszentrum Jülich a time resolution of about 80 ps has been achieved using SiPMs from KETEK and Hamamatsu with an active area of 3 × 3mm2. Employing the Digital Photon Counter from Philips a time resolution of about 30 ps has been reached.

  3. Optimization of Stability, Encapsulation, Release, and Cross-Priming of Tumor Antigen-Containing PLGA Nanoparticles

    PubMed Central

    Prasad, Shashi; Cody, Virginia; Saucier-Sawyer, Jennifer K.; Fadel, Tarek R.; Edelson, Richard L.; Birchall, Martin A.

    2014-01-01

    Purpose In order to investigate Poly (lactic-co-glycolic acid) (PLGA) nanoparticles (NP) as potential vehicles for efficient tumor antigen (TA) delivery to dendritic cells (DC), this study aimed to optimize encapsulation/release kinetics before determining immunogenicity of antigen-containing NP. Methods Various techniques were used to liberate TA from cell lines. Single (gp100) and multiple (B16-tumor lysate containing gp100) antigens were encapsulated within differing molecular weight PLGA co-polymers. Differences in morphology, encapsulation/release and biologic potency were studied. Findings were adopted to encapsulate fresh tumor lysate from patients with advanced tumors and compare stimulation of tumor infiltrating lymphocytes (TIL) against that achieved by soluble lysate. Results Four cycles of freeze-thaw + 15 s sonication resulted in antigen-rich lysates without the need for toxic detergents or protease inhibitors. The 80KDa polymer resulted in maximal release of payload and favorable production of immunostimulatory IL-2 and IFN-γ. NP-mediated antigen delivery led to increased IFN-γ and decreased immunoinhibitory IL-10 synthesis when compared to soluble lysate. Conclusions Four cycles of freeze-thaw followed by 15 s sonication is the ideal technique to obtain complex TA for encapsulation. The 80KDa polymer has the most promising combination of release kinetics and biologic potency. Encapsulated antigens are immunogenic and evoke favorable TIL-mediated anti-tumor responses. PMID:22798259

  4. A comparative study on genetic effects of artificial and natural habitat fragmentation on Loropetalum chinense (Hamamelidaceae) in Southeast China

    PubMed Central

    Yuan, N; Comes, H P; Cao, Y N; Guo, R; Zhang, Y H; Qiu, Y X

    2015-01-01

    Elucidating the demographic and landscape features that determine the genetic effects of habitat fragmentation has become fundamental to research in conservation and evolutionary biology. Land-bridge islands provide ideal study areas for investigating the genetic effects of habitat fragmentation at different temporal and spatial scales. In this context, we compared patterns of nuclear microsatellite variation between insular populations of a shrub of evergreen broad-leaved forest, Loropetalum chinense, from the artificially created Thousand-Island Lake (TIL) and the Holocene-dated Zhoushan Archipelago of Southeast China. Populations from the TIL region harboured higher levels of genetic diversity than those from the Zhoushan Archipelago, but these differences were not significant. There was no correlation between genetic diversity and most island features, excepting a negative effect of mainland–island distance on allelic richness and expected heterozygosity in the Zhoushan Archipelago. In general, levels of gene flow among island populations were moderate to high, and tests of alternative models of population history strongly favoured a gene flow-drift model over a pure drift model in each region. In sum, our results showed no obvious genetic effects of habitat fragmentation due to recent (artificial) or past (natural) island formation. Rather, they highlight the importance of gene flow (most likely via seed) in maintaining genetic variation and preventing inter-population differentiation in the face of habitat ‘insularization' at different temporal and spatial scales. PMID:25515015

  5. Thermal safety characterization and explosion violence of energetic materials

    NASA Astrophysics Data System (ADS)

    Hsu, Peter; Hust, Gary; Pagoria, Philip; Fried, Larry

    2015-06-01

    Some energetic materials could thermally explode at fairly low temperatures (<100 C) and the violence from thermal explosion may cause a significant damage. Thus understanding the response of energetic material to thermal events is very important for the storage and handling of energetic materials. Over the last few decades, there has been considerable research effort on the thermal decomposition and thermal explosion violence of energetic materials at elevated temperatures in different sample geometries and confinements. Among them, the ODTX system is an interesting option due to its sample requirement and easiness for data modeling. It has been used since 1970s for cook-off study at LLNL. It generates 3 technical data: (1) lowest temperature at which thermal explosion would occur (threshold temperature, Til) , (2) times to thermal explosion at temperature above Til, for the calculation of activation energy and frequency factor; and (3) thermal explosion violence. In this paper, we will present some recent ODTX experimental data of several new energetic materials as well as gas pressure data at elevated temperature.

  6. A comparative study on genetic effects of artificial and natural habitat fragmentation on Loropetalum chinense (Hamamelidaceae) in Southeast China.

    PubMed

    Yuan, N; Comes, H P; Cao, Y N; Guo, R; Zhang, Y H; Qiu, Y X

    2015-06-01

    Elucidating the demographic and landscape features that determine the genetic effects of habitat fragmentation has become fundamental to research in conservation and evolutionary biology. Land-bridge islands provide ideal study areas for investigating the genetic effects of habitat fragmentation at different temporal and spatial scales. In this context, we compared patterns of nuclear microsatellite variation between insular populations of a shrub of evergreen broad-leaved forest, Loropetalum chinense, from the artificially created Thousand-Island Lake (TIL) and the Holocene-dated Zhoushan Archipelago of Southeast China. Populations from the TIL region harboured higher levels of genetic diversity than those from the Zhoushan Archipelago, but these differences were not significant. There was no correlation between genetic diversity and most island features, excepting a negative effect of mainland-island distance on allelic richness and expected heterozygosity in the Zhoushan Archipelago. In general, levels of gene flow among island populations were moderate to high, and tests of alternative models of population history strongly favoured a gene flow-drift model over a pure drift model in each region. In sum, our results showed no obvious genetic effects of habitat fragmentation due to recent (artificial) or past (natural) island formation. Rather, they highlight the importance of gene flow (most likely via seed) in maintaining genetic variation and preventing inter-population differentiation in the face of habitat 'insularization' at different temporal and spatial scales.

  7. Metastasis is regulated via microRNA-200/ZEB1 axis control of tumor cell PD-L1 expression and intratumoral immunosuppression

    PubMed Central

    Goswami, Sangeeta; Cortez, Maria Angelica; Ahn, Young-Ho; Byers, Lauren A.; Zhang, Xuejun; Yi, Xiaohui; Dwyer, David; Lin, Wei; Diao, Lixia; Wang, Jing; Roybal, Jonathon; Patel, Mayuri; Ungewiss, Christin; Peng, David; Antonia, Scott; Mediavilla-Varela, Melanie; Robertson, Gordon; Suraokar, Milind; Welsh, James W.; Erez, Baruch; Wistuba, Ignacio I.; Chen, Lieping; Peng, Di; Wang, Shanshan; Ullrich, Stephen E.; Heymach, John V.; Kurie, Jonathan M.; Qin, F. Xiao-Feng

    2014-01-01

    Immunosuppression of tumor-infiltrating lymphocytes (TIL) is a common feature of advanced cancer, but its biological basis has remained obscure. We demonstrate here a molecular link between epithelial-to-mesenchymal transition (EMT) and CD8+ TIL immunosuppression, two key drivers of cancer progression. We show that microRNA-200 (miR-200), a cell-autonomous suppressor of EMT and metastasis, targets PD-L1. Moreover, ZEB1, an EMT activator and transcriptional repressor of miR-200, relieves miR-200 repression of PD-L1 on tumor cells, leading to CD8+ T cell immunosuppression and metastasis. These findings are supported by robust correlations between the EMT score, miR-200 levels and PD-L1 expression in multiple human lung cancer datasets. In addition to revealing a link between EMT and T cell dysfunction, these findings also show that ZEB1 promotes metastasis through a heretofore unappreciated cell non-autonomous mechanism, and suggest that subgroups of patients in whom malignant progression is driven by EMT activators may respond to treatment with PD-L1 antagonists. PMID:25348003

  8. HHLA2, a member of the B7 family, is expressed in human osteosarcoma and is associated with metastases and worse survival

    PubMed Central

    Koirala, Pratistha; Roth, Michael E.; Gill, Jonathan; Chinai, Jordan M.; Ewart, Michelle R.; Piperdi, Sajida; Geller, David S.; Hoang, Bang H.; Fatakhova, Yekaterina V.; Ghorpade, Maya; Zang, Xingxing; Gorlick, Richard

    2016-01-01

    Over the past four decades there have been minimal improvements in outcomes for patients with osteosarcoma. New targets and novel therapies are needed to improve outcomes for these patients. We sought to evaluate the prevalence and clinical significance of the newest immune checkpoint, HHLA2, in osteosarcoma. HHLA2 protein expression was evaluated in primary tumor specimens and metastatic disease using an osteosarcoma tumor microarray (TMA) (n = 62). The association of HHLA2 with the presence of tumor infiltrating lymphocytes (TILs) and five-year-event-free-survival were examined. HHLA2 was expressed in 68% of osteosarcoma tumors. HHLA2 was expressed in almost all metastatic disease specimens and was more prevalent than in primary specimens without known metastases (93% vs 53%, p = 0.02). TILs were present in 75% of all osteosarcoma specimens. Patients whose tumors were ≥25% or ≥50% HHLA2 positive had significantly worse five-year event-free-survival (33% vs 64%, p = 0.03 and 14% vs 59%, p = 0.02). Overall, we have shown that HHLA2 is expressed in the majority of osteosarcoma tumors and its expression is associated with metastatic disease and poorer survival. Along with previously reported findings that HHLA2 is a T cell co-inhibitor, these results suggest that HHLA2 may be a novel immunosuppressive mechanism within the osteosarcoma tumor microenvironment. PMID:27531281

  9. Stereospecificity of isotopic exchange of C-α-protons of glycine catalyzed by three PLP-dependent lyases: the unusual case of tyrosine phenol-lyase.

    PubMed

    Koulikova, Vitalia V; Zakomirdina, Lyudmila N; Gogoleva, Olga I; Tsvetikova, Marina A; Morozova, Elena A; Komissarov, Vsevolod V; Tkachev, Yaroslav V; Timofeev, Vladimir P; Demidkina, Tatyana V; Faleev, Nicolai G

    2011-11-01

    A comparative study of the kinetics and stereospecificity of isotopic exchange of the pro-2R- and pro-2S protons of glycine in (2)H(2)O under the action of tyrosine phenol-lyase (TPL), tryptophan indole-lyase (TIL) and methionine γ-lyase (MGL) was undertaken. The kinetics of exchange was monitored using both (1)H- and (13)C-NMR. In the three compared lyases the stereospecificities of the main reactions with natural substrates dictate orthogonal orientation of the pro-2R proton of glycine with respect to the cofactor pyridoxal 5'-phosphate (PLP) plane. Consequently, according to Dunathan's postulate with all the three enzymes pro-2R proton should exchange faster than does the pro-2S one. In fact the found ratios of 2R:2S reactivities are 1:20 for TPL, 108:1 for TIL, and 1,440:1 for MGL. Thus, TPL displays an unprecedented inversion of stereospecificity. A probable mechanism of the observed phenomenon is suggested, which is based on the X-ray data for the quinonoid intermediate, formed in the reaction of TPL with L-alanine. The mechanism implies different conformational changes in the active site upon binding of glycine and alanine. These changes can lead to relative stabilization of either the neutral amino group, accepting the α-proton, or the respective ammonium group, which is formed after the proton abstraction.

  10. Regulatory T cells, especially ICOS+ FOXP3+ regulatory T cells, are increased in the hepatocellular carcinoma microenvironment and predict reduced survival

    PubMed Central

    Tu, Jian-Fei; Ding, Ya-Hui; Ying, Xi-Hui; Wu, Fa-Zong; Zhou, Xin-Mu; Zhang, Deng-Ke; Zou, Hai; Ji, Jian-Song

    2016-01-01

    Hepatocellular carcinoma (HCC) is a common malignant tumour, especially in Asia. Its prognosis is poor, and there are limited methods for predicting patient survival. This study was carried out to analyse the prognostic value of tumour-infiltrating lymphocytes (TILs), especially regulatory T cells (Tregs), in HCC patients. TILs were analysed in 57 randomly selected HCC patients. The prognostic effects of groups with high and low numbers were evaluated by the Kaplan-Meier and Cox model analyses. Although higher densities of CD3+, CD4+, and CD8+ cytotoxic lymphocytes (CTLs) as well as CD56+ NK cells and CD68+ macrophages were observed in peritumoural tissue, increased numbers of forkhead/winged helix transcription factor P3+ (FOXP3+) Tregs were found in intratumoural tissue. Additionally, regarding ICOS+ FOXP3+ Tregs, an increased prevalence in carcinoma was not only associated with the absolute number but also with the percentage of FOXP3+ cells. Higher Treg levels in tumour tissues indicated a worse prognosis, and the FOXP3+ Tregs/CD4+ T cells ratio was an independent prognostic factor for OS. Therefore, FOXP3+ Tregs, especially ICOS+ FOXP3+ Tregs, contribute to the immunosuppressive HCC microenvironment. High tumour-infiltrating Tregs are thought to be an unfavourable prognostic indicator of HCC. PMID:27725696

  11. Correlation between anti-PD-L1 tumor concentrations and tumor-specific and nonspecific biomarkers in a melanoma mouse model.

    PubMed

    Contreras-Sandoval, Ana M; Merino, María; Vasquez, Marcos; Trocóniz, Iñaki F; Berraondo, Pedro; Garrido, María J

    2016-11-22

    Blockade of PD-L1 with specific monoclonal antibodies (anti-PD-L1) represents a therapeutic strategy to increase the capability of the immune system to modulate the tumor immune-resistance. The relationship between anti-PD-L1 tumor exposition and anti-tumor effect represents a challenge that has been addressed in this work through the identification of certain biomarkers implicated in the antibody's mechanism of action, using a syngeneic melanoma mouse model. The development of an in-vitro/in-vivo platform has allowed us to investigate the PD-L1 behavior after its blockage with anti-PD-L1 at cellular level and in animals. In-vitro studies showed that the complex PD-L1/anti-PD-L1 was retained mainly at the cell surface. The antibody concentration and time exposure affected directly the recycling or ligand turnover. In-vivo studies showed that anti-PD-L1 was therapeutically active at all stage of the disease, with a rapid onset, a low but durable efficacy and non-relevant toxic effect. This efficacy measured as tumor shrinkage correlated with tumor-specific infiltrating lymphocytes (TILs), which increased as antibody tumor concentrations increased. Both, TILS and antibody concentrations followed similar kinetic patterns, justifying the observed anti-PD-L1 rapid onset. Interestingly, peripheral lymphocytes (PBLs) behave as infiltrating lymphocytes, suggesting that these PBLs might be considered as a possible biomarker for antibody activity.

  12. Crucial Contributions by T Lymphocytes (Effector, Regulatory, and Checkpoint Inhibitor) and Cytokines (TH1, TH2, and TH17) to a Pathological Complete Response Induced by Neoadjuvant Chemotherapy in Women with Breast Cancer

    PubMed Central

    Verma, Chandan; Eremin, Jennifer M.; Cowley, Gerard; Ilyas, Mohammed; Eremin, Oleg

    2016-01-01

    The tumour microenvironment consists of malignant cells, stroma, and immune cells. Prominent tumour-infiltrating lymphocytes (TILs) in breast cancer are associated with a good prognosis and are predictors of a pathological complete response (pCR) with neoadjuvant chemotherapy (NAC). The contribution of different T effector/regulatory cells and cytokines to tumour cell death with NAC requires further characterisation and was investigated in this study. Breast tumours from 33 women with large and locally advanced breast cancers undergoing NAC were immunohistochemically (intratumoural, stromal) assessed for T cell subsets and cytokine expression using labelled antibodies, employing established semiquantitative methods. Prominent levels of TILs and CD4+, CD8+, and CTLA-4+ (stromal) T cells and CD8+ : FOXP3+ ratios were associated with a significant pCR; no association was seen with FOXP3+, CTLA-4+ (intratumoural), and PD-1+ T cells. NAC significantly reduced CD4+, FOXP3+, CTLA-4+ (stromal) (concurrently blood FOXP3+, CTLA-4+ Tregs), and PD-1+ T cells; no reduction was seen with CD8+ and CTLA-4+ (intratumoural) T cells. High post-NAC tumour levels of FOXP3+ T cells, IL-10, and IL-17 were associated with a failed pCR. Our study has characterised further the contribution of T effector/regulatory cells and cytokines to tumour cell death with NAC. PMID:27777963

  13. Temperature-controlled ionic liquid-based ultrasound-assisted microextraction for preconcentration of trace quantity of cadmium and nickel by using organic ligand in artificial saliva extract of smokeless tobacco products.

    PubMed

    Arain, Sadaf Sadia; Kazi, Tasneem Gul; Arain, Asma Jabeen; Afridi, Hassan Imran; Baig, Jameel Ahmed; Brahman, Kapil Dev; Naeemullah; Arain, Salma Aslam

    2015-03-05

    A new approach was developed for the preconcentration of cadmium (Cd) and nickel (Ni) in artificial saliva extract of dry snuff (brown and black) products using temperature-controlled ionic liquid-based ultrasound-assisted dispersive liquid-liquid microextraction (TIL-UDLLμE) followed by electrothermal atomic absorption spectrometry (ETAAS). The Cd and Ni were complexed with ammonium pyrrolidinedithiocarbamate (APDC), extracted in ionic liquid drops, 1-butyl-3-methylimidazolium hexafluorophosphate [C4MIM][PF6]. The multivariate strategy was applied to estimate the optimum values of experimental variables influence the % recovery of analytes by TIL-UDLLμE method. At optimum experimental conditions, the limit of detection (3s) were 0.05 and 0.14μgL(-1) while relative standard deviations (% RSD) were 3.97 and 3.55 for Cd and Ni respectively. After extraction, the enhancement factors (EF) were 87 and 79 for Cd and Ni, respectively. The RSD for six replicates of 10μgL(-1) Cd and Ni were 3.97% and 3.55% respectively. To validate the proposed method, certified reference material (CRM) of Virginia tobacco leaves was analyzed, and the determined values of Cd and Ni were in good agreement with the certified values. The concentration of Cd and Ni in artificial saliva extracts corresponds to 39-52% and 21-32%, respectively, of the total contents of both elements in dry brown and black snuff products.

  14. Effects of Urechis unicinctus juveniles on chemical characteristics of organically contaminated coastal sediment

    NASA Astrophysics Data System (ADS)

    Kang, Kyoung Ho; Zhang, Zhifeng; Kim, Jae Min; Seon, Seung Chun; Shao, Mingyu

    2010-03-01

    Biological activities of marine benthos such as burrowing and feeding may change sediment characteristics. We conducted three experiments to examine the potential of using juveniles of a spoon worm Urechis unicinctus to improve the quality of organically contaminated coastal sediment. Sediment samples were collected from a site that was heavily contaminated with organic matter (Seonso) and two sites that were clean (Myo-do, Dolsan-do). Urechis juveniles, obtained by artificial fertilization and cultured in the laboratory, were introduced to the sediment (weight 3 kg, depth 10 cm) at a density of 500 individuals per aquarium (length 50 cm, width 35 cm, height 30 cm) (Experiment 1), or at densities ranging from 100 to 900 individuals per beaker (Experiment 2). To examine how sediment contamination can be modified by the effects of Urechis, 500 individuals (per aquarium) were exposed to the Seonso contaminated sediment that had been mixed with 0-100% clean sand (Experiment 3). Each experiment lasted two months and sediment samples were collected every 15 d to determine the several indexes of sediment quality, which included acid volatile sulfide (AVS), chemical oxygen demand (COD) and total ignition loss (TIL). In Experiment 1, the existence of Urechis did not result in significant changes in quality indexes in the sediments collected from Myo-do, Dolsan-do. However, AVS, COD and TIL of the Seonso sediment all decreased significantly after co-incubation with Urechis juveniles for 30 to 45 d. Experiment 2 showed that a density of at least 300 juveniles per beaker was necessary to significantly reduce all three quality indexes, and the magnitude of reduction was positively correlated with juvenile density. Experiment 3 revealed that Urechis juveniles were effective in reducing the AVS, COD and TIL of the Seonso sediment that had been mixed with 60%, 80%, and 80% of clean sand, respectively. The results of the present study therefore indicated that juveniles of this

  15. Distribution of immune cells in head and neck cancer: CD8+ T-cells and CD20+ B-cells in metastatic lymph nodes are associated with favourable outcome in patients with oro- and hypopharyngeal carcinoma

    PubMed Central

    2009-01-01

    Background Tumour infiltrating lymphocytes (TIL) are generally considered to represent a host immune response directed against tumour antigens. TIL are also increasingly recognised as possible prognostic parameters. However, the effects observed are variable indicating that results cannot be extrapolated from type of tumour to another. Moreover, it has been suggested that primary solid tumours may be ignored by the immune system and that a meaningful immune response is only mounted in regional lymph nodes. Methods We have examined the local distribution of immune cells in tumour-related compartments in head and neck squamous cell carcinomas (HNSCC). In a second step, the prognostic impact of these cells on disease-free survival (DFS) was analysed. A total of 198 tissue cores from 33 patients were evaluated using tissue mircroarray technique and immunohistochemistry. Tumour-infiltrating immune cells were identified using antibodies specific for CD3, CD8, GranzymeB, FoxP3, CD20 and CD68 and quantified using an image analysis system. Results We demonstrate a relative expansion of FoxP3+ regulatory T-cells (Treg) and of cytotoxic T-cells among tumour infitrating T-cells. We also show that intratumoural CD20+ B-cells are significantly more frequent in metastatic deposits than in primary tumours. Furthermore, we observed a reduced number of peritumoural CD8+ T-cells in metastatic lymph nodes as compared to univolved regional nodes suggesting a local down-modulation of cellular immunity. All other immune cells did not show significant alterations in distribution. We did not observe an association of tumour infiltrating immune cells at the primary site with outcome. However, increased numbers of intraepithelial CD8+ TIL in metastatic tumours as well as large numbers of peritumoural B-cells in lymph node metastases were associated with favourable outcome. Unexpectedly, no effect on patient outcome was observed for Treg in any compartment. Conclusion Our results suggest that

  16. Variants in autophagy-related genes and clinical characteristics in melanoma: a population-based study.

    PubMed

    White, Kirsten A M; Luo, Li; Thompson, Todd A; Torres, Salina; Hu, Chien-An Andy; Thomas, Nancy E; Lilyquist, Jenna; Anton-Culver, Hoda; Gruber, Stephen B; From, Lynn; Busam, Klaus J; Orlow, Irene; Kanetsky, Peter A; Marrett, Loraine D; Gallagher, Richard P; Sacchetto, Lidia; Rosso, Stefano; Dwyer, Terence; Cust, Anne E; Begg, Colin B; Berwick, Marianne

    2016-11-01

    Autophagy has been linked with melanoma risk and survival, but no polymorphisms in autophagy-related (ATG) genes have been investigated in relation to melanoma progression. We examined five single-nucleotide polymorphisms (SNPs) in three ATG genes (ATG5; ATG10; and ATG16L) with known or suspected impact on autophagic flux in an international population-based case-control study of melanoma. DNA from 911 melanoma patients was genotyped. An association was identified between (GG) (rs2241880) and earlier stage at diagnosis (OR 0.47; 95% Confidence Intervals (CI) = 0.27-0.81, P = 0.02) and a decrease in Breslow thickness (P = 0.03). The ATG16L heterozygous genotype (AG) (rs2241880) was associated with younger age at diagnosis (P = 0.02). Two SNPs in ATG5 were found to be associated with increased stage (rs2245214 CG, OR 1.47; 95% CI = 1.11-1.94, P = 0.03; rs510432 CC, OR 1.84; 95% CI = 1.12-3.02, P = 0.05). Finally, we identified inverse associations between ATG5 (GG rs2245214) and melanomas on the scalp or neck (OR 0.20, 95% CI = 0.05-0.86, P = 0.03); ATG10 (CC) (rs1864182) and brisk tumor infiltrating lymphocytes (TILs) (OR 0.42; 95% CI = 0.21-0.88, P = 0.02), and ATG5 (CC) (rs510432) with nonbrisk TILs (OR 0.55; 95% CI = 0.34-0.87, P = 0.01). Our data suggest that ATG SNPs might be differentially associated with specific host and tumor characteristics including age at diagnosis, TILs, and stage. These associations may be critical to understanding the role of autophagy in cancer, and further investigation will help characterize the contribution of these variants to melanoma progression.

  17. Assessing somatic hypermutation in Ramos B cells after overexpression or knockdown of specific genes.

    PubMed

    Upton, Dana C; Unniraman, Shyam

    2011-11-01

    B cells start their life with low affinity antibodies generated by V(D)J recombination. However, upon detecting a pathogen, the variable (V) region of an immunoglobulin (Ig) gene is mutated approximately 100,000-fold more than the rest of the genome through somatic hypermutation (SHM), resulting in high affinity antibodies. In addition, class switch recombination (CSR) produces antibodies with different effector functions depending on the kind of immune response that is needed for a particular pathogen. Both CSR and SHM are initiated by activation-induced cytidine deaminase (AID), which deaminates cytosine residues in DNA to produce uracils. These uracils are processed by error-prone forms of repair pathways, eventually leading to mutations and recombination. Our current understanding of the molecular details of SHM and CSR come from a combination of studies in mice, primary cells, cell lines, and cell-free experiments. Mouse models remain the gold standard with genetic knockouts showing critical roles for many repair factors (e.g. Ung, Msh2, Msh6, Exo1, and polymerase η). However, not all genes are amenable for knockout studies. For example, knockouts of several double-strand break repair proteins are embryonically lethal or impair B-cell development. Moreover, sometimes the specific function of a protein in SHM or CSR may be masked by more global defects caused by the knockout. In addition, since experiments in mice can be lengthy, altering expression of individual genes in cell lines has become an increasingly popular first step to identifying and characterizing candidate genes. Ramos - a Burkitt lymphoma cell line that constitutively undergoes SHM - has been a popular cell-line model to study SHM. One advantage of Ramos cells is that they have a built-in convenient semi-quantitative measure of SHM. Wild type cells express IgM and, as they pick up mutations, some of the mutations knock out IgM expression. Therefore, assaying IgM loss by fluorescence

  18. Delayed sleep phase syndrome in adolescents: prevalence and correlates in a large population based study

    PubMed Central

    2013-01-01

    Background The aims of this study were to estimate the prevalence of Delayed Sleep Phase Syndrome (DSPS) in adolescence, and to examine the association to insomnia and school non-attendance. Methods Data stem from a large population based study in Hordaland County in Norway conducted in 2012, the ung@hordaland study. In all, 10,220 adolescents aged 16–18 years (54% girls) provided self-reported data on a range of sleep parameters: DSPS was defined according to the International Classification of Sleep Disorders, Revised (ICSD-R) criteria, while insomnia was defined according to the Quantitative Criteria for Insomnia. Other sleep parameters included time in bed, sleep duration, sleep efficiency, oversleeping, sleep onset latency, wake after sleep onset, subjective sleep need, sleep deficiency, sleepiness and tiredness. Sleep data were calculated separately for weekdays and weekends. Data on school non-attendance were provided by official registers. Results The prevalence of DSPS was 3.3%, and significantly higher among girls (3.7%) than boys (2.7%). There was a strong overlap between DSPS and insomnia, with more than half of the adolescents with DSPS also meeting the criteria for insomnia (53.8% for boys and 57.1% for girls). Adolescents with DSPS had significantly higher odds ratios (OR) of non-attendance at school. After adjusting for sociodeographical factors, insomnia and depression, the adjusted ORs for days of non-attendance were OR = 3.22 (95% CI: 1.94-5.34) for boys and OR = 1.87 (95% CI: 1.25-2.80) for girls. A similar effect was found for hours of non-attendance for boys, with an adjusted OR = 3.05 (95% CI: 1.89-4.92). The effect for girls was no longer significant after full adjustment (OR =1.48 [95% CI: 0.94-2.32]). Conclusions This is one of the first studies to estimate the prevalence of DSPS in adolescents. The high prevalence of DSPS, and overlap with insomnia, in combination with the odds of school non-attendance, suggest that a broad

  19. HDAC Inhibitors Upregulate B Cell microRNAs that Silence AID and Blimp-1 Expression for Epigenetic Modulation of Antibody and Autoantibody Responses

    PubMed Central

    White, Clayton A; Pone, Egest J; Lam, Tonika; Tat, Connie; Hayama, Ken L; Li, Guideng; Zan, Hong; Casali, Paolo

    2014-01-01

    Class-switch DNA recombination (CSR) and somatic hypermutation (SHM), which require AID, and plasma cell differentiation, which requires Blimp-1, are critical for the generation of class-switched and hypermutated (mature) antibody and autoantibody responses. We showed here that the histone deacetylase (HDAC) inhibitors (HDI) valproic acid (VPA) and butyrate upregulated miR-155, miR-181b and miR-361, which silenced AICDA/Aicda (AID) mRNA, and miR-23b, miR-30a and miR-125b, which silenced PRDM1/Prdm1 (Blimp-1) mRNA, in human and mouse B cells. This led to downregulation of AID, Blimp-1 and Xbp-1 expression, thereby dampening CSR, SHM and plasma cell differentiation without altering B cell viability or proliferation. The selectivity of HDI-mediated silencing of AICDA/Aicda and PRDM1/Prdm1 was emphasized by unchanged expression of HoxC4 and Irf4 (important inducers/modulators of AICDA/Aicda), Rev1 and Ung (central elements for CSR/SHM), and Bcl6, Bach2 or Pax5 (repressors of PRDM1/Prdm1 expression), as well as unchanged expression of miR-19a/b, miR-20a and miR-25, which are not known to regulate AICDA/Aicda or PRDM1/Prdm1. Through these B cell intrinsic epigenetic mechanisms, VPA blunted class-switched and hypermutated T-dependent and T-independent antibody responses in C57BL/6 mice. In addition, it decreased class-switched and hypermutated autoantibodies, ameliorated disease and extended survival in lupus MRL/Faslpr/lpr mice. Our findings outline epigenetic mechanisms that modulate expression of an enzyme (AID) and transcription factors (Blimp-1 and Xbp-1) that critical to the B cell differentiation processes that underpin antibody and autoantibody responses. They also provide therapeutics proof-of-principle in autoantibody-mediated autoimmunity. PMID:25392531

  20. OSRD Reports: Bibliography and Index of Declassified Reports Having OSRD Numbers

    DTIC Science & Technology

    1947-06-01

    lillti. tD HI ,’.00 Ph ".00. OSRD 116 ’BL QI)f (I-SO. toftlllt. J. L, WId OIadoel1, "arrl. M. 11 .ortUl. phllta for til. ,roduetlOft of liquid 0...1071) H .... leo. Jr. ..,de.ta for , .. lIted Uk_ff of ,1,,1 ., • .; t"’l r: u .. .,d th. ,rtYllltion 0’ ,I .. t Injury tD the ,1,,1.,.. ’ro...34lty. I".Utul, of OpUc. ’ Ald. tD "IIIIIt vl.ton: EJt.ft.𔃺II of the _topic yl.lblllt1. ClInt to atSQA and to IOODA. Ita, 1M!. ’SO dr ... I

  1. NERF - A Computer Program for the Numerical Evaluation of Reliability Functions - Reliability Modelling, Numerical Methods and Program Documentation,

    DTIC Science & Technology

    1983-09-01

    no.t, tl J)’HS( ( (%-no)t/tl j +nO-,d )tij/ Ais (nO+(,&-no)tij/t). ...... (2.52) For given no, all structures with _no1-(td-no)t/til have been removed by...the reliability functions is that the inverse crack growth function is available, i.e., t - a’ 1 (a) (3.21) for at, a&af, where ai -a(Ti) and afb- a( t...r I ( t)ZI I P no¢ ni)P.C€ .in/f (noft)lRRn/#f(nott) nOet, not t) .,R -t I(R3 ~oet 3T3 mi n X ai ny"( Zt) ( .T 3 which leads to the expression for

  2. Word Frequency Analysis MOS: 71M. Skill Levels 1 & 2.

    DTIC Science & Technology

    1981-05-01

    2016 P»3E P> 9 O e D C o •j o 2 a^c ">n t ut-.li ro I Ul -A l...S032S 2016 *HGE cruNT« vc»a^ 3 EXTENSION 2 f’XTPC": 4 FACCS I* F..C1LITY 9 F,.IK 1 fCB 16 FICLJ • til il 1 • it Flies 1... 2016 PAGE CCJN*4 WCRD* I l«T’EP.SP 1 I’ll" ? I ’VIT1NG 1 UMK V IIW MS •) ll’rmr, 4 ItWGKH « IF’TNG JuV’iJ Hi i ill i I I • ’ •

  3. Storm Induced Changes of the Topside Ionosphere as Deduced from Incoherent Scatter Radars

    DTIC Science & Technology

    1990-01-01

    11484 Sep 18 05 16 to 1q84 Sep1, 9 05.16 UT Millstone Hill Ion teOMPO’dture (Til (K 2 4 6 8 10 12 14 16 18 20 22 24 LT 600 550 (b) j 500 450 350 b B 10 2...l 350 320 L/ 2 4 6 8 10 12 Id 16. )s 20 22 24 -1 1q84 Oct 17 00 08 o 19)84 Oct 18 00 08 UT Arecibo 22 24 2 4 6 8 0 12 14 16 8 20L 600 I (b) 450 400Y...22 24 UT 19B4 Sep iq 00.08 to 1q84 Sop 20 eZ.OB UT Arecibo Ion, tomp.o-atw-. IT1 K 1 22 24 2 4 6 8 10 12 14 16 18 20 LT 550 500J.50 (b) 45 400 350

  4. Mapping titanium and tin oxide phases using EELS: an application of independent component analysis.

    PubMed

    de la Peña, F; Berger, M-H; Hochepied, J-F; Dynys, F; Stephan, O; Walls, M

    2011-01-01

    We study materials that present challenges for conventional elemental mapping techniques and can in some cases be treated successfully using independent component analysis (ICA). In this case the material in question is obtained from a TiO₂-SiO₂ solid solution that is spinodally decomposed into TiO₂ rich-SnO₂ rich multilayers. Conventional elemental mapping is difficult because the edges most easily mapped for these elements (Ti-L, Sn-M and O-K) all have onsets within the same 80 eV range. ICA is used to separate entire spectral signals corresponding to particular material phases or molecular units rather than particular elements and is thus able to distinguish between TiO₂ and SnO₂. We show that quantification of oxide species can be performed by different methods that require extra assumptions, but nevertheless should be feasible in many cases.

  5. Intravital imaging of multicolor-labeled tumor immune microenvironment through skin-fold window chamber

    NASA Astrophysics Data System (ADS)

    Qi, Shuhong; Zhang, Zhihong

    2015-03-01

    Tumor immune microenvironment became very important for the tumor immunotherapy. There were several kinds of immune cells in tumor stromal, and they played very different roles in tumor growth. In order to observe the behaviors of multiple immune cells in tumor microenvironment and the interaction between immune cells and tumor cells at the same time, we generated a multicolor-labeled tumor immune microenvironment model. The tumor cells and immune cells were labeled by different fluorescent proteins. By using of skin-fold window chamber implanted into mice and intravital imaging technology, we could dynamically observe the different immune cells in tumor microenvironment. After data analysis from the video, we could know the behavior of TILs, DCs and Tregs in tumor immune microenvironment; furthermore, we could know these immune cells play different roles in the tumor microenvironment.

  6. Modeling of Shadows in Radar Clutter.

    DTIC Science & Technology

    1980-07-01

    0005 wAITE 46.20001 TITLE 060 WRITE 46.21331 N!.4V.NC.NTIN.N4TOUT.M4TIRL.1PAUtF.NETNGO 0007 N1.1 0006 4 2WNI*NCONV 0000 N43 .42 * PIC 0010 NA-43#HE 0011...PARTMtON PARAM7!E C EPpTI. SEP.) PCtNTS ARE %fLPCTEfW FQROS THE 0ATA UNITS. c PEA) TM" SEOU.INCF NUftERS POrN TeE CHOSEN 0A74 cjo ItSOPOPM A"OfS so2 IN...34 MCM WASIARLE FOR Iwo, CLU6Tal 00*?0 00 M.0TMI 006444 gu. 910V 006? so sOMITIN C TilPt PIC W e"RijiUO 0011 CPIOTRJI TOTALIJI 1/00141014JI 60S4Go o ca its

  7. Tall oil as additive in gas drive hydrocarbon oil recovery

    SciTech Connect

    Djabbarah, N.F.

    1988-04-12

    A miscible displacement process for recovering oil from a subterranean, oil-containing formation penetrated by at least one injection well and at least one spaced-apart production well and having fluid communication between the injection and the production wells is described comprising: (a) injecting a slug of til oil into the formation through the injection well; (b) injecting a slug of a displacing fluid into the formation through the injection well, the displacing fluid being selected from the group consisting of carbon monoxide, carbon dioxide, methane, nitrogen, air, flue gas, combustion gas and mixtures thereof, the injection of the tall oil lowering the minimum miscibility pressure of the displacing fluid in the formation oil; and (c) recovering the oil through the production well.

  8. [Genetic information from tumor-infiltrating B lymphocytes as a driver tool ("GPS") for anti-tumor T cell CARs].

    PubMed

    Kotlan, Beatrix; Csuka, Orsolya; Tóth, László; Farkas, Emil; Plótár, Vanda; Horváth, Szabolcs; Éles, Klára; Olasz, Judit; Tóth, József; Kásler, Miklós; Liszkay, Gabriella

    2016-03-02

    The rapidly growing field of gene therapy techniques to modify T cells with chimeric antigen receptors (CARs) for cancer care solutions, reached considerable achievements. However, there is an urgent need of reliable, well tolerable tumor-associated antigen specific antibodies. Tumor-infiltrating B (TIL-B) cell originated single chain Fv (scFv) gene regions could be selected with tumor specificity. DNA sequences of these antibody variable regions were subjects to get engineered into new CAR constructs. Our novel strategy harnesses tumor-infiltrating B cells' unique capacity to reveal highly tumor-associated disialylated glycosphingolipids (GD3 gangliosides). We used these human antibody fragments for generating GD3 ganglioside specific CAR gene constructs for potential usage in solid tumors.

  9. U.S. Atlantic Command Exercise Cooperative Safeguard 1997 (CS-97) After Action Report

    DTIC Science & Technology

    1997-10-01

    m t^ t^ CO CO CO a o O o o s in m (O r~ i~- CM CM CM CM CM 01 X <o m E3ro 1A-: til:: lO- H ifl> Lfl:- H tn: WMwWwM i in IO m in in fi.i\\ tu...tn ct in tn :° |^ = H :0 |cn io :*H : 1 iO ■o ■o Ct o m in in ._ o CD in •-4 m •-I v4 i c ■ « ■ fc JCQ i c ■•H...Thomas Hall Dir. Naval Reserve Association Ms Nan Borton Dir. Office of Foreign Disaster Assistance Dr. Clair Blong Dir. Of International Programs

  10. Ammonium Perchlorate Transportation Hazards Testing

    DTIC Science & Technology

    1989-04-01

    n1 t- TI T1W- r- T’ T II 0 0L 10 T. J II )0 In L11 n n i) ) C, (71 T-. d n 𔃺 ’ O 4j ’G134j L) ) 0 4j .1-10. 0 Il I-I 60 In 0 InI ni ni nini r , - r...n Q TIT a tf M L) QtQ0 O 4 4 0.14 3 41,31)-1 17 �’tIl 014 l’ 01 i. 0 - a. OC t. L 0I n nI nini n0ni nu n- n-r- rd Ifl 0n - 44 4 0o 3-00- 300- -TI

  11. Computer Graphics Research Laboratory Quarterly Progress Report Number 49, July-September 1993

    DTIC Science & Technology

    1993-11-22

    foj’ e f ",0, " Wl oIMOn of 6flnOmatl.0. q~ mttmatel(a tO fa d 1q no 0.* -Ae i.o’ ncluding thN t.me w NYll.1O’ Wae.ctuni e .4.inq "ato ’.Okdt" qaute...Ad na n* nt.flq I-P Gala -"eede and co4Iwelatqn ade r@...*nq thpoec.ti~lon of ,MOUtft.Ofl %end tooWnkM’ti tidatnq in"i awler",f P"t’~. e a’ 4ny...other aWed of tk.n coiloct Onal o1nlfl,.nat,0P fl..(o.afq ’.uqqe..on’. to, eaucintq til’. Guidoe- to *44.n qton .oeaom.alite.%’ . e Dfeedle. for IntOIAt’O

  12. Word Criticality Analysis MOS: 13F. Skill Level 1 and 2.

    DTIC Science & Technology

    1981-09-01

    W.’ s PP060. 12’?p PFGE 3 13#AMSW41SSY(Rs 7~ ,F 2-182.1I 2- 1241*I TSKIKY- P1231/6 -2-131I v &tC. 2-309,2 2-308,1 2-107,1 2-222.2 2-14.9,1 2- 11.5 2...0000 0 ON IfCFILE COPY HEADQUARTERS DATA CONTflL 11 UNITED STATES ARMY TRAINING AND DOCTRINE COMMAND FORT MONROE, VIRGINIA 23651 " S \\, / UA 4 E.U,~ I...I 77 = FTM-,~ ’T M777 i_ - ’. UM9CLW~IFIED SECUMTT CLASSIFICATION OF THIS PACE (1W.. 001. Bntwnq REPORT DOCUMENTATION PAGE 83I OM~m 4TIL(adA~daa S

  13. New Insights into the Role of the Immune Microenvironment in Breast Carcinoma

    PubMed Central

    de la Cruz-Merino, Luis; Barco-Sánchez, Antonio; Henao Carrasco, Fernando; Nogales Fernández, Esteban; Vallejo Benítez, Ana; Brugal Molina, Javier; Martínez Peinado, Antonio; Grueso López, Ana; Ruiz Borrego, Manuel; Codes Manuel de Villena, Manuel; Sánchez-Margalet, Víctor; Nieto-García, Adoración; Alba Conejo, Emilio; Casares Lagar, Noelia; Ibáñez Martínez, José

    2013-01-01

    Recently, immune edition has been recognized as a new hallmark of cancer. In this respect, some clinical trials in breast cancer have reported imppressive outcomes related to laboratory immune findings, especially in the neoadjuvant and metastatic setting. Infiltration by tumor infiltrating lymphocytes (TIL) and their subtypes, tumor-associated macrophages (TAM) and myeloid-derived suppressive cells (MDSC) seem bona fide prognostic and even predictive biomarkers, that will eventually be incorporated into diagnostic and therapeutic algorithms of breast cancer. In addition, the complex interaction of costimulatory and coinhibitory molecules on the immune synapse and the different signals that they may exert represent another exciting field to explore. In this review we try to summarize and elucidate these new concepts and knowledge from a translational perspective focusing on breast cancer, paying special attention to those aspects that might have more significance in clinical practice and could be useful to design successful therapeutic strategies in the future. PMID:23861693

  14. Liquid Nitrogen (Oxygen Simulant) Thermodynamic Vent System Test Data Analysis

    NASA Technical Reports Server (NTRS)

    Hedayat, A.; Nelson, S. L.; Hastings, L. J.; Flachbart, R. H.; Tucker, S. P.

    2005-01-01

    In designing systems for the long-term storage of cryogens in low-gravity (space) environments, one must consider the effects of thermal stratification on tank pressure that will occur due to environmental heat leaks. During low-gravity operations, a Thermodynamic Vent System (TVS) concept is expected to maintain tank pressure without propellant resettling. A series of TVS tests was conducted at NASA Marshall Space Flight Center (MSFC) using liquid nitrogen (LN2) as a liquid oxygen (LO2) simulant. The tests were performed at tank til1 levels of 90%, 50%, and 25%, and with a specified tank pressure control band. A transient one-dimensional TVS performance program is used to analyze and correlate the test data for all three fill levels. Predictions and comparisons of ullage pressure and temperature and bulk liquid saturation pressure and temperature with test data are presented.

  15. Integrated Data Analysis (IDCA) Program - PETN Class 4 Standard

    SciTech Connect

    Sandstrom, Mary M.; Brown, Geoffrey W.; Preston, Daniel N.; Pollard, Colin J.; Warner, Kirstin F.; Sorensen, Daniel N.; Remmers, Daniel L.; Shelley, Timothy J.; Reyes, Jose A.; Phillips, Jason J.; Hsu, Peter C.; Reynolds, John G.

    2012-08-01

    The Integrated Data Collection Analysis (IDCA) program is conducting a proficiency study for Small- Scale Safety and Thermal (SSST) testing of homemade explosives (HMEs). Described here are the results for impact, friction, electrostatic discharge, and differential scanning calorimetry analysis of PETN Class 4. The PETN was found to have: 1) an impact sensitivity (DH50) range of 6 to 12 cm, 2) a BAM friction sensitivity (F50) range 7 to 11 kg, TIL (0/10) of 3.7 to 7.2 kg, 3) a ABL friction sensitivity threshold of 5 or less psig at 8 fps, 4) an ABL ESD sensitivity threshold of 0.031 to 0.326 j/g, and 5) a thermal sensitivity of an endothermic feature with Tmin = ~ 141 °C, and a exothermic feature with a Tmax = ~205°C.

  16. Fasting-Mimicking Diet Reduces HO-1 to Promote T Cell-Mediated Tumor Cytotoxicity.

    PubMed

    Di Biase, Stefano; Lee, Changhan; Brandhorst, Sebastian; Manes, Brianna; Buono, Roberta; Cheng, Chia-Wei; Cacciottolo, Mafalda; Martin-Montalvo, Alejandro; de Cabo, Rafael; Wei, Min; Morgan, Todd E; Longo, Valter D

    2016-07-11

    Immune-based interventions are promising strategies to achieve long-term cancer-free survival. Fasting was previously shown to differentially sensitize tumors to chemotherapy while protecting normal cells, including hematopoietic stem and immune cells, from its toxic side effects. Here, we show that the combination of chemotherapy and a fasting-mimicking diet (FMD) increases the levels of bone marrow common lymphoid progenitor cells and cytotoxic CD8(+) tumor-infiltrating lymphocytes (TILs), leading to a major delay in breast cancer and melanoma progression. In breast tumors, this effect is partially mediated by the downregulation of the stress-responsive enzyme heme oxygenase-1 (HO-1). These data indicate that FMD cycles combined with chemotherapy can enhance T cell-dependent targeted killing of cancer cells both by stimulating the hematopoietic system and by enhancing CD8(+)-dependent tumor cytotoxicity.

  17. PREFACE: XXII International Conference on Integrable Systems and Quantum Symmetries (ISQS-22)

    NASA Astrophysics Data System (ADS)

    2014-11-01

    The XXII International Conference on Integrable Systems and Quantum symmetries (ISQS-22), organized by the Department of Mathematics, Faculty of Nuclear Sciences and Physical Engineering, Czech Technical University Prague and the Bogoliubov Laboratory of Theoretical Physics of the Joint Institute for Nuclear Research, belongs to the successful series of conferences held at the Czech Technical University which began in 1992 and is devoted to problems of mathematical physics related to the theory of integrable systems, quantum groups and quantum symmetries. During the last 20 years, each of the conferences gathered around 60 scientists from all over the world. 33 papers of plenary lectures and contributions presented at ISQS-22 are published in the present issue of Journal of Physics: Conference Series. Čestmír Burdík, Ondřej Navrátil, Severin Pošta Editors

  18. FOREWORD: XXth International Conference on Integrable Systems and Quantum Symmetries (ISQS-20)

    NASA Astrophysics Data System (ADS)

    Burdík, Čestmír; Navrátil, Ondřej; Pošta, Severin

    2013-01-01

    The XXth International Conference on Integrable Systems and Quantum Symmetries (ISQS-20), organized by the Department of Mathematics, Faculty of Nuclear Sciences and Physical Engineering, Czech Technical University, Prague, and the Bogoliubov Laboratory of Theoretical Physics of the Joint Institute for Nuclear Research, is part of the successful series of conferences held at the Czech Technical University which began in 1992, and is devoted to problems of mathematical physics related to the theory of integrable systems, quantum groups and quantum symmetries. During the last 20 years each of these conferences has gathered around 60 scientists from all over the world. 31 plenary lectures and contributions presented at ISQS-20 are published in this volume of Journal of Physics: Conference Series. Guest Editors Čestmír Burdík, Ondřej Navrátil and Severin Pošta

  19. US Army Human Engineering Laboratory Communications Survey-A Pilot Study (HELCOMS-PS).

    DTIC Science & Technology

    1984-11-01

    AD-Ai5i 434 US ARMY HUMAN ENGINEERING LABORATORY COMMUNICATIONS i/I. SURVEY-A PILOT STUDY (HELCOMS- PS )(U) RRMAMENT SYSTEMS INC ABERDEEN ND R M PHELPS...P gITT)Y ( HIE1JY)i1- PS S VtW A 1 ,’ ii ’ M TT))’ jn kj I DTI itprov, f or ’ , 1 19𔃿 , ’t r ih),I t ni-til re m l " U. S. ARMY HUMAN ENGINEERING...Final % VE-A P {1,0T STUDY (HEI :OMS- PS ) Y PERFORMING ORG. REPORT N, MREIR 7 A 8 CONTRACT OR GRANT NUMHLFP-,|j1 ,I (ell 𔃻. Phelps ( eorfi- A. Kupets, Sr

  20. USAF/SCEEE Graduate Student Summer Support Program (1982). Management and Technical Report.

    DTIC Science & Technology

    1982-10-01

    and testing of a nitric oxide Raman cell wavelength shifter is described. This device is used in conjunction with a Q-switched Nd:Glass laser system, to...ignment~ , i-l r-tit’? -t a nvtric oxid ,- Rami-n 1. -.,.%o-t-,lth l -i ft er ; 4o r;c iI4. m qv!;*, t t l, ai vt 1, ;gb n e ak no’.or qI r irl pmht pTIL... odifiCations can be easily made since the model has captured the essence of the problem. With the val id~ited rl.,del thle sinalybt can Lreate enough

  1. Evaluation of Alumina Ceramic Housings for Deep Submergence Service. Fifth Generation Housings. Part 1

    DTIC Science & Technology

    1994-03-01

    consolida- assembly. Similarly, eight hemispheres were iso - tion process results in net shrinkage of the ceramic statically pressed to yield the two...I Cr)- * .5 ! C * 0 Cic I " 45I (ID II C . LC m Ir o 0 0 0 , Ire, II -j ILJ z . U I - 4 46 I IM I CA * 09 .4 m Iso Po I I C -~LL iii II w, HLn0 00...013369 0 1X1279 0 19-1,. 0 ( 13333 0.Ŕ:38 0119I5 W 11(61527 0 (IM-19 0 (100899 (OX)40, It19(9tI-l, 11 I4561o - - )1110N4115 - - 111)(2710 - - I 11 L3

  2. Strategies to modulate the immune system in breast cancer: checkpoint inhibitors and beyond

    PubMed Central

    Migali, Cristina; Milano, Monica; Trapani, Dario; Criscitiello, Carmen; Esposito, Angela; Locatelli, Marzia; Minchella, Ida; Curigliano, Giuseppe

    2016-01-01

    Is breast cancer (BC) immunogenic? Many data suggest that it is. Many observations demonstrated the prognostic role of tumor-infiltrating lymphocytes (TILs) in triple negative (TN) and human epidermal growth factor receptor 2 (HER-2)-positive BC. TNBCs are poorly differentiated tumors with high genetic instability and very high heterogeneity. This heterogeneity enhances the ‘danger signals’ and select clone variants that could be more antigenic or, in other words, that could more strongly stimulate a host immune antitumor response. The response to chemotherapy is at least partly dependent on an immunological reaction against those tumor cells that are dying during the chemotherapy. One of the mechanisms whereby chemotherapy can stimulate the immune system to recognize and destroy malignant cells is commonly known as immunogenic cell death (ICD). ICD elicits an adaptive immune response. Which are the clinical implications of all ‘immunome’ data produced in the last years? First, validate prognostic or predictive role of TILs. Second, validate immune genomic signatures that may be predictive and prognostic in patients with TN disease. Third, incorporate an ‘immunoscore’ into traditional classification of BC, thus providing an essential prognostic and potentially predictive tool in the pathology report. Fourth, implement clinical trials for BC in the metastatic setting with drugs that target immune-cell–intrinsic checkpoints. Blockade of one of these checkpoints, cytotoxic T-lymphocyte-associated antigen-4 (CTLA-4) or the programmed cell death 1 (PD-1) receptor may provide proof of concepts for the activity of an immune-modulation approach in the treatment of a BC. PMID:27583028

  3. The Identification of Congeners and Aliens by Drosophila Larvae.

    PubMed

    Del Pino, Francisco; Jara, Claudia; Pino, Luis; Medina-Muñoz, María Cristina; Alvarez, Eduardo; Godoy-Herrera, Raúl

    2015-01-01

    We investigated the role of Drosophila larva olfactory system in identification of congeners and aliens. We discuss the importance of these activities in larva navigation across substrates, and the implications for allocation of space and food among species of similar ecologies. Wild type larvae of cosmopolitan D. melanogaster and endemic D. pavani, which cohabit the same breeding sites, used species-specific volatiles to identify conspecifics and aliens moving toward larvae of their species. D. gaucha larvae, a sibling species of D. pavani that is ecologically isolated from D. melanogaster, did not respond to melanogaster odor cues. Similar to D. pavani larvae, the navigation of pavani female x gaucha male hybrids was influenced by conspecific and alien odors, whereas gaucha female x pavani male hybrid larvae exhibited behavior similar to the D. gaucha parent. The two sibling species exhibited substantial evolutionary divergence in processing the odor inputs necessary to identify conspecifics. Orco (Or83b) mutant larvae of D. melanogaster, which exhibit a loss of sense of smell, did not distinguish conspecific from alien larvae, instead moving across the substrate. Syn97CS and rut larvae of D. melanogaster, which are unable to learn but can smell, moved across the substrate as well. The Orco (Or83b), Syn97CS and rut loci are necessary to orient navigation by D. melanogaster larvae. Individuals of the Trana strain of D. melanogaster did not respond to conspecific and alien larval volatiles and therefore navigated randomly across the substrate. By contrast, larvae of the Til-Til strain used larval volatiles to orient their movement. Natural populations of D. melanogaster may exhibit differences in identification of conspecific and alien larvae. Larval locomotion was not affected by the volatiles.

  4. Tumor-infiltrating NY-ESO-1-specific CD8+ T cells are negatively regulated by LAG-3 and PD-1 in human ovarian cancer.

    PubMed

    Matsuzaki, Junko; Gnjatic, Sacha; Mhawech-Fauceglia, Paulette; Beck, Amy; Miller, Austin; Tsuji, Takemasa; Eppolito, Cheryl; Qian, Feng; Lele, Shashikant; Shrikant, Protul; Old, Lloyd J; Odunsi, Kunle

    2010-04-27

    NY-ESO-1 is a "cancer-testis" antigen frequently expressed in epithelial ovarian cancer (EOC) and is among the most immunogenic tumor antigens defined to date. In an effort to understand in vivo tolerance mechanisms, we assessed the phenotype and function of NY-ESO-1-specific CD8(+) T cells derived from peripheral blood lymphocytes (PBLs), tumor-infiltrating lymphocytes (TILs), and tumor-associated lymphocytes (TALs) of EOC patients with NY-ESO-1-expressing tumors, with or without humoral immunity to NY-ESO-1. Whereas NY-ESO-1-specific CD8(+) T cells were readily detectable ex vivo with tetramers in TILs and TALs of seropositive patients, they were only detectable in PBLs following in vitro stimulation. Compared with PBLs, tumor-derived NY-ESO-1-specific CD8(+) T cells demonstrated impaired effector function, preferential usage of dominant T-cell receptor, and enriched coexpression of inhibitory molecules LAG-3 and PD-1. Expression of LAG-3 and PD-1 on CD8(+) T cells was up-regulated by IL-10, IL-6 (cytokines found in tumor ascites), and tumor-derived antigen-presenting cells. Functionally, CD8(+)LAG-3(+)PD-1(+) T cells were more impaired in IFN-gamma/TNF-alpha production compared with LAG-3(+)PD-1(-) or LAG-3(-)PD-1(-) subsets. Dual blockade of LAG-3 and PD-1 during T-cell priming efficiently augmented proliferation and cytokine production by NY-ESO-1-specific CD8(+) T cells, indicating that antitumor function of NY-ESO-1-specific CD8(+) T cells could potentially be improved by therapeutic targeting of these inhibitory receptors.

  5. Plastid Proteomic Analysis in Tomato Fruit Development

    PubMed Central

    Kondo, Takanori; Dohra, Hideo; Ito, Yumihiko; Kiriiwa, Yoshikazu; Hayashi, Marina; Kamiya, Shiori; Kato, Masaya; Fujiwara, Masayuki; Fukao, Yoichiro; Kobayashi, Megumi; Nagata, Noriko; Motohashi, Reiko

    2015-01-01

    To better understand the mechanism of plastid differentiation from chloroplast to chromoplast, we examined proteome and plastid changes over four distinct developmental stages of ‘Micro-Tom’ fruit. Additionally, to discover more about the relationship between fruit color and plastid differentiation, we also analyzed and compared ‘Micro-Tom’ results with those from two other varieties, ‘Black’ and ‘White Beauty’. We confirmed that proteins related to photosynthesis remain through the orange maturity stage of ‘Micro-Tom’, and also learned that thylakoids no longer exist at this stage. These results suggest that at a minimum there are changes in plastid morphology occurring before all related proteins change. We also compared ‘Micro-Tom’ fruits with ‘Black’ and ‘White Beauty’ using two-dimensional gel electrophoresis. We found a decrease of CHRC (plastid-lipid-associated protein) and HrBP1 (harpin binding protein-1) in the ‘Black’ and ‘White Beauty’ varieties. CHRC is involved in carotenoid accumulation and stabilization. HrBP1 in Arabidopsis has a sequence similar to proteins in the PAP/fibrillin family. These proteins have characteristics and functions similar to lipocalin, an example of which is the transport of hydrophobic molecules. We detected spots of TIL (temperature-induced lipocalin) in 2D-PAGE results, however the number of spots and their isoelectric points differed between ‘Micro-Tom’ and ‘Black’/‘White Beauty’. Lipocalin has various functions including those related to environmental stress response, apoptosis induction, membrane formation and fixation, regulation of immune response, cell growth, and metabolism adjustment. Lipocalin related proteins such as TIL and HrBP1 could be related to the accumulation of carotenoids, fruit color and the differentiation of chromoplast. PMID:26371478

  6. Generation of autologous tumor-specific T cells for adoptive transfer based on vaccination, in vitro restimulation and CD3/CD28 dynabead-induced T cell expansion.

    PubMed

    Brimnes, Marie Klinge; Gang, Anne Ortved; Donia, Marco; Thor Straten, Per; Svane, Inge Marie; Hadrup, Sine Reker

    2012-08-01

    Adoptive cell transfer (ACT) of in vitro expanded autologous tumor-infiltrating lymphocytes (TIL) has been shown to exert therapeutic efficacy in melanoma patients. We aimed to develop an ACT protocol based on tumor-specific T cells isolated from peripheral blood and in vitro expanded by Dynabeads® ClinExVivo™CD3/CD28. We show here that the addition of an in vitro restimulation step with relevant peptides prior to bead expansion dramatically increased the proportion of tumor-specific T cells in PBMC-cultures. Importantly, peptide-pulsed dendritic cells (DCs) as well as allogeneic tumor lysate-pulsed DCs from the DC vaccine preparation could be used with comparable efficiency to peptides for in vitro restimulation, to increase the tumor-specific T-cell response. Furthermore, we tested the use of different ratios and different types of Dynabeads® CD3/CD28 and CD3/CD28/CD137 T-cell expander, for optimized expansion of tumor-specific T cells. A ratio of 1:3 of Dynabeads® CD3/CD28 T-cell expander to T cells resulted in the maximum number of tumor-specific T cells. The addition of CD137 did not improve functionality or fold expansion. Both T-cell expansion systems could generate tumor-specific T cells that were both cytotoxic and effective cytokine producers upon antigen recognition. Dynabeads®-expanded T-cell cultures shows phenotypical characteristics of memory T cells with potential to migrate and expand in vivo. In addition, they possess longer telomeres compared to TIL cultures. Taken together, we demonstrate that in vitro restimulation of tumor-specific T cells prior to bead expansion is necessary to achieve high numbers of tumor-specific T cells. This is effective and easily applicable in combination with DC vaccination, by use of vaccine-generated DCs, either pulsed with peptide or tumor-lysate.

  7. Temperature controlled ionic liquid-based dispersive micro-extraction using two ligands, for determination of aluminium in scalp hair samples of Alzheimer's patients: A multivariate study

    NASA Astrophysics Data System (ADS)

    Arain, Mariam S.; Arain, Salma A.; Kazi, Tasneem G.; Afridi, Hassan I.; Ali, Jamshaid; Naeemulllah; Arain, Sadaf S.; Brahman, Kapil Dev; Mughal, Moina Akhtar

    2015-02-01

    A green and sensitive temperature controlled dispersive liquid-liquid microextraction (TIL-DLLME) methodology based on the application of ionic liquid 1-butyl-3-methylimidazolium hexafluorophosphate, [C4mim][PF6], as an extractant solvent was proposed for the preconcentration of trace levels of aluminium (Al3+) in scalp hair samples of Alzheimer's (AD) patients, prior to analyzing by flame atomic absorption spectrometry (FAAS). The Al3+ was complexed with 8-hydrooxyquinoline (oxine) (L1) and 3,5,7,2‧-4‧ pentahydroxy flavone (morin) (L2) separately and then extracted by IL at temperature (50 ± 2.0 °C). Some effective factors that influence the TIL-DLLME efficiency such as pH, ligands concentrations, volume of IL, ionic strength, and incubation time were investigated and optimized by multivariate analysis. In the optimum experimental conditions, the limit of detection (3 s) and enhancement factor were 0.56 μg L-1, 0.64 μg L-1 and 85, 73 for both ligands, respectively. The relative standard deviation (RSD) for six replicate determinations of 100 μg L-1 Al3+ complexed with oxine and morin were found to be 3.88% and 4.74%, respectively. The developed method was validated by the analysis of certified reference material of human hair (NCSZC81002).and applied satisfactorily to the determination of Al3+ in acid digested scalp hair samples of AD patients and healthy controls. The resulted data shows significant higher level in scalp hair samples of AD male patients with related to referents of same age and socioeconomic status.

  8. The sialotranscriptome of Antricola delacruzi female ticks is compatible with non-hematophagous behavior and an alternative source of food

    PubMed Central

    Ribeiro, José Marcos C.; Labruna, Marcelo B.; Mans, Ben J.; Maruyama, Sandra Regina; Francischetti, Ivo M. B.; Barizon, Gustavo Canavaci; de Miranda Santos, Isabel K. F.

    2012-01-01

    The hosts for Antricola delacruzi ticks are insectivorous, cave-dwelling bats on which only larvae are found. The mouthparts of nymphal and adult A. delacruzi are compatible with scavenging feeding because the hypostome is small and toothless. How a single blood meal of a larva provides energy for several molts as well as for oviposition by females is not known. Adults of A. delacruzi possibly feed upon an unknown food source in bat guano, a substrate on which nymphal and adult stages are always found. Guano produced by insectivorous bats contains twice the amount of protein and 60 times the amount of iron as beef. In addition, bacteria and chitin-rich fungi proliferate on guano. Comparative data on the transcriptome of the salivary glands of A. delacruzi is nonexistent and would help to understand the physiological adaptations of salivary glands that accompany different sources of food as well as the steps taken by the Acari towards haematophagy, believed to have evolved from scavenging dead animals. Annotation of the transcriptome of salivary glands from female instars of A. delacruzi collected on guano categorized 5.7% of the clusters of expressed genes as putative secreted proteins. They included abundantly expressed TIL domain-containing proteins (possible anti-microbials), an abundantly expressed protein similar to a serum amyloid found in the sialotranscriptomes of Ornithodoros spp., a savignygrin, a family of mucin/peritrophin/cuticle-like proteins, antimicrobials and an HIV envelope-like glycoprotein also found in soft ticks. When comparing the transcriptome of A. delacruzi with those of blood-feeding female soft and hard ticks some notable differences were observed; they consisted of the following transcripts over- or under-represented or absent in the sialotranscriptome of A. delacuzi that may reflect its source of food: ferritin, mucins with chitin-binding domains and TIL domain-containing proteins versus lipocalins, basic tail proteins

  9. Induced Pluripotent Stem Cells: Challenges and Opportunities for Cancer Immunotherapy

    PubMed Central

    Sachamitr, Patty; Hackett, Simon; Fairchild, Paul Jonathan

    2014-01-01

    Despite recent advances in cancer treatment over the past 30 years, therapeutic options remain limited and do not always offer a cure for malignancy. Given that tumor-associated antigens (TAA) are, by definition, self-proteins, the need to productively engage autoreactive T cells remains at the heart of strategies for cancer immunotherapy. These have traditionally focused on the administration of autologous monocyte-derived dendritic cells (moDC) pulsed with TAA, or the ex vivo expansion and adoptive transfer of tumor-infiltrating lymphocytes (TIL) as a source of TAA-specific cytotoxic T cells (CTL). Although such approaches have shown some efficacy, success has been limited by the poor capacity of moDC to cross present exogenous TAA to the CD8+ T-cell repertoire and the potential for exhaustion of CTL expanded ex vivo. Recent advances in induced pluripotency offer opportunities to generate patient-specific stem cell lines with the potential to differentiate in vitro into cell types whose properties may help address these issues. Here, we review recent success in the differentiation of NK cells from human induced pluripotent stem (iPS) cells as well as minor subsets of dendritic cells (DCs) with therapeutic potential, including CD141+XCR1+ DC, capable of cross presenting TAA to naïve CD8+ T cells. Furthermore, we review recent progress in the use of TIL as the starting material for the derivation of iPSC lines, thereby capturing their antigen specificity in a self-renewing stem cell line, from which potentially unlimited numbers of naïve TAA-specific T cells may be differentiated, free of the risks of exhaustion. PMID:24860566

  10. Characterizing the immune microenvironment of malignant peripheral nerve sheath tumor by PD-L1 expression and presence of CD8+ tumor infiltrating lymphocytes

    PubMed Central

    Shurell, Elizabeth; Singh, Arun S.; Crompton, Joseph G.; Jensen, Sarah; Li, Yunfeng; Dry, Sarah; Nelson, Scott; Chmielowski, Bartosz; Bernthal, Nicholas; Federman, Noah; Tumeh, Paul; Eilber, Fritz C.

    2016-01-01

    Background Malignant peripheral nerve sheath tumor (MPNST) is an aggressive sarcoma with few treatment options. Tumor immune state has not been characterized in MPNST, and is important in determining response to immune checkpoint blockade. Our aim was to evaluate the expression of programmed death-ligand 1 (PD-L1), programmed cell death protein 1 (PD-1), and presence of CD8+ tumor infiltrating lymphocytes (TILs) in MPNST, and correlate these findings with clinical behavior and outcome. Results PD-L1 staining of at least 1% was seen in 0/20 nerves, 2/68 benign lesions and 9/53 MPNST. Two of 68 benign lesions and 7/53 (13%) MPNST had at least 5% PD-L1 staining. CD8 staining of at least 5% was seen in 1/20 (5%) nerves, 45/68 (66%) benign lesions and 30/53 (57%) MPNST. PD-L1 was statistically more prevalent in MPNST than both nerves and benign lesions (p=0.049 and p=0.008, respectively). Expression of PD-1 was absent in all tissue specimens. There was no correlation of PD-L1 or CD8 expression with disease state (primary versus metastatic) or patient survival. Methods A comprehensive PNST tissue microarray was created from 141 surgical specimens including primary, recurrent, and metastatic MPNST (n=53), neurofibromas (n=57), schwannoma (n=11), and normal nerve (n=20). Cores were stained in triplicate for PD-L1, PD-1, and CD8, and expression compared between tumor types. These data were then examined for survival correlates in 35 patients with primary MPNST. Conclusions MPNST is characterized by low PD-L1 and absent PD-1 expression with significant CD8+ TIL presence. MPNST immune microenvironment does not correlate with patient outcome. PMID:27588404

  11. PD-1 expression conditions T cell avidity within an antigen-specific repertoire

    PubMed Central

    Simon, Sylvain; Vignard, Virginie; Florenceau, Laetitia; Dreno, B.; Khammari, A.; Lang, F.; Labarriere, N.

    2016-01-01

    ABSTRACT Despite its negative regulatory role on tumor-specific T cells, Programmed cell death 1 (PD-1) is also a marker of activated tumor-infiltrating T cells. In cancer, PD-1 blockade partially reverses T cell dysfunction allowing the amplification of tumor reactive T cells. Here, we investigated the role of PD-1 signaling on effector/memory human T cells specific for shared melanoma antigens, derived from blood. We documented for the first time the existence of melanoma-specific T cell clones unable to express PD-1. This stable feature was due to the persistent methylation of the PDCD1 promoter. These PD-1neg clones were of lower avidity than their PD-1pos counterparts, suggesting that high-affinity-specific T cell clones unable to express PD-1 are not or rarely present in peripheral blood, as they are probably eliminated by negative selection, due to their high reactivity. We also documented the existence of such PD-1neg T cell clones in melanoma tumor-infiltrating lymphocytes (TIL), which also exhibited a lower functional avidity than PD-1pos TIL clones. This clearly shows that PD-1 expression identifies antigen-specific T cell clonotypes of high functional avidity. Finally, we demonstrated that PD-1 blockade during the in vitro selection process of Melan-A-specific T cells favored the amplification of higher avidity T cell clonotypes. This preferential amplification of high-avidity memory T cells upon PD-1 blockade resonates with the expansion of reactive T cells, including neo-antigen-specific T cells observed in anti-PD-1-treated patients. This feature should also be a useful biomarker of clinical efficiency, while providing new insights for adoptive transfer treatments. PMID:26942093

  12. Importance of CD200 expression by tumor or host cells to regulation of immunotherapy in a mouse breast cancer model.

    PubMed

    Curry, Anna; Khatri, Ismat; Kos, Olha; Zhu, Fang; Gorczynski, Reginald

    2017-01-01

    Cell-surface CD200 expression by mouse EMT6 breast tumor cells increased primary tumor growth and metastasis to the draining lymph nodes (DLN) in normal (WT) BALB/c female recipients, while lack of CD200R1 expression in a CD200R1-/- host negated this effect. Silencing CD200 expression in EMT6siCD200 tumor cells also reduced their ability to grow and metastasize in WT animals. The cellular mechanisms responsible for these effects have not been studied in detail. We report characterization of tumor infiltrating (TILs) and draining lymph node (DLN) cells in WT and CD200-/- BALB/c mice, receiving WT tumor cells, or EMT6 lacking CD200 expression (EMT6siCD200 cells). Our data show an important correlation with augmented CD8+ cytotoxic T cells and resistance to tumor growth in mice lacking exposure (on either host cells or tumor) to the immunoregulatory molecule CD200. Confirmation of the importance of such CD8+ cells came from monitoring tumor growth and characterization of the TILs and DLN cells in WT mice challenged with EMT6 and EMT6siCD200 tumors and treated with CD8 and CD4 depleting antibodies. Finally, we have assessed the mechanisms(s) whereby addition of metformin as an augmenting chemotherapeutic agent in CD200-/- animals given EMT6 tumors and treated with a previously established immunotherapy regime can increase host resistance. Our data support the hypothesis that increased autophagy in the presence of metformin increases CD8+ responses and tumor resistance, an effect attenuated by the autophagy inhibitor verteporfin.

  13. Clonal expansion of antitumor T cells in breast cancer correlates with response to neoadjuvant chemotherapy

    PubMed Central

    Park, Jae-Hyun; Jang, Miran; Tarhan, Yunus Emre; Katagiri, Toyomasa; Sasa, Mitsunori; Miyoshi, Yasuo; Kalari, Krishna R.; Suman, Vera J.; Weinshilboum, Richard; Wang, Liewei; Boughey, Judy C.; Goetz, Matthew P.; Nakamura, Yusuke

    2016-01-01

    The immune microenvironment of tumor plays a critical role in therapeutic responses to chemotherapy. Cancer tissues are composed of a complex network between anti-tumor and pro-tumor immune cells and molecules; therefore a comprehensive analysis of the tumor immune condition is imperative for better understanding of the roles of the immune microenvironment in anticancer treatment response. In this study, we performed T cell receptor (TCR) repertoire analysis of tumor infiltrating T cells (TILs) in cancer tissues of pre- and post-neoadjuvant chemotherapy (NAC) from 19 breast cancer patients; five cases showed CR (complete response), ten showed PR (partial response), and four showed SD/PD (stable disease/progressive disease) to the treatment. From the TCR sequencing results, we calculated the diversity index of the TCRβ chain and found that clonal expansion of TILs could be detected in patients who showed CR or PR to NAC. Noteworthy, the diversity of TCR was further reduced in the post-NAC tumors of CR patients. Our quantitative RT-PCR also showed that expression ratio of CD8/Foxp3 was significantly elevated in the post-NAC tumors of CR cases (p=0.0032), indicating that antitumor T cells were activated and enriched in these tumors. Collectively, our findings suggest that the clonal expansion of antitumor T cells may be a critical factor associated with response to chemotherapy and that their TCR sequences might be applicable for the development of TCR-engineered T cells treatment for individual breast cancer patients when their tumors relapse. PMID:27278091

  14. Genome-wide identification and immune response analysis of serine protease inhibitor genes in the silkworm, Bombyx mori.

    PubMed

    Zhao, Ping; Dong, Zhaoming; Duan, Jun; Wang, Genhong; Wang, Lingyan; Li, Youshan; Xiang, Zhonghuai; Xia, Qingyou

    2012-01-01

    In most insect species, a variety of serine protease inhibitors (SPIs) have been found in multiple tissues, including integument, gonad, salivary gland, and hemolymph, and are required for preventing unwanted proteolysis. These SPIs belong to different families and have distinct inhibitory mechanisms. Herein, we predicted and characterized potential SPI genes based on the genome sequences of silkworm, Bombyx mori. As a result, a total of eighty SPI genes were identified in B. mori. These SPI genes contain 10 kinds of SPI domains, including serpin, Kunitz_BPTI, Kazal, TIL, amfpi, Bowman-Birk, Antistasin, WAP, Pacifastin, and alpha-macroglobulin. Sixty-three SPIs contain single SPI domain while the others have at least two inhibitor units. Some SPIs also contain non-inhibitor domains for protein-protein interactions, including EGF, ADAM_spacer, spondin_N, reeler, TSP_1 and other modules. Microarray analysis showed that fourteen SPI genes from lineage-specific TIL family and Group F of serpin family had enriched expression in the silk gland. The roles of SPIs in resisting pathogens were investigated in silkworms when they were infected by four pathogens. Microarray and qRT-PCR experiments revealed obvious up-regulation of 8, 4, 3 and 3 SPI genes after infection with Escherichia coli, Bacillus bombysepticus, Beauveria bassiana or B. mori nuclear polyhedrosis virus (BmNPV), respectively. On the contrary, 4, 11, 7 and 9 SPI genes were down-regulated after infection with E. coli, B. bombysepticus, B. bassiana or BmNPV, respectively. These results suggested that these SPI genes may be involved in resistance to pathogenic microorganisms. These findings may provide valuable information for further clarifying the roles of SPIs in the development, immune defence, and efficient synthesis of silk gland protein.

  15. Compensatory upregulation of PD-1, LAG-3, and CTLA-4 limits the efficacy of single-agent checkpoint blockade in metastatic ovarian cancer.

    PubMed

    Huang, Ruea-Yea; Francois, Ariel; McGray, Aj Robert; Miliotto, Anthony; Odunsi, Kunle

    2017-01-01

    Tumor-associated or -infiltrating lymphocytes (TALs or TILs) co-express multiple immune inhibitory receptors that contribute to immune suppression in the ovarian tumor microenvironment (TME). Dual blockade of PD-1 along with LAG-3 or CTLA-4 has been shown to synergistically enhance T-cell effector function, resulting in a delay in murine ovarian tumor growth. However, the mechanisms underlying this synergy and the relative contribution of other inhibitory receptors to immune suppression in the ovarian TME are unknown. Here, we report that multiple immune checkpoints are expressed in TALs and TILs isolated from ovarian tumor-bearing mice. Importantly, blockade of PD-1, LAG-3, or CTLA-4 alone using genetic ablation or blocking antibodies conferred a compensatory upregulation of the other checkpoint pathways, potentiating their capacity for local T-cell suppression that, in turn, could be overcome through combinatorial blockade strategies. Whereas single-agent blockade led to tumor outgrowth in all animals, dual antibody blockade against PD-1/CTLA-4 or triple blockade against PD-1/LAG-3/CTLA-4 resulted in tumor-free survival in 20% of treated mice. In contrast, dual blockade of LAG-3 and CTLA-4 pathways using PD-1 knockout mice led to tumor-free survival in 40% of treated mice, suggesting a hierarchical ordering of checkpoint function. Durable antitumor immunity was most strongly associated with increased numbers of CD8(+) T cells, the frequency of cytokine-producing effector T cells, reduced frequency of Tregs and arginine-expressing monocytic myeloid-derived suppressor cells in the peritoneal TME. These data provide a basis for combinatorial checkpoint blockade in clinical intervention for ovarian cancer.

  16. Cytokine profiling of tumor interstitial fluid of the breast and its relationship with lymphocyte infiltration and clinicopathological characteristics.

    PubMed

    Espinoza, Jaime A; Jabeen, Shakila; Batra, Richa; Papaleo, Elena; Haakensen, Vilde; Timmermans Wielenga, Vera; Møller Talman, Maj-Lis; Brunner, Nils; Børresen-Dale, Anne-Lise; Gromov, Pavel; Helland, Åslaug; Kristensen, Vessela N; Gromova, Irina

    2016-01-01

    The tumor microenvironment is composed of many immune cell subpopulations and is an important factor in the malignant progression of neoplasms, particularly breast cancer (BC). However, the cytokine networks that coordinate various regulatory events within the BC interstitium remain largely uncharacterized. Moreover, the data obtained regarding the origin of cytokine secretions, the levels of secretion associated with tumor development, and the possible clinical relevance of cytokines remain controversial. Therefore, we profiled 27 cytokines in 78 breast tumor interstitial fluid (TIF) samples, 43 normal interstitial fluid (NIF) samples, and 25 matched serum samples obtained from BC patients with Luminex xMAP multiplex technology. Eleven cytokines exhibited significantly higher levels in the TIF samples compared with the NIF samples: interleukin (IL)-7, IL-10, fibroblast growth factor-2, IL-13, interferon (IFN)γ-inducible protein (IP-10), IL-1 receptor antagonist (IL-1RA), platelet-derived growth factor (PDGF)-β, IL-1β, chemokine ligand 5 (RANTES), vascular endothelial growth factor, and IL-12. An immunohistochemical analysis further demonstrated that IL-1RA, IP-10, IL-10, PDGF-β, RANTES, and VEGF are widely expressed by both cancer cells and tumor-infiltrating lymphocytes (TILs), whereas IP-10 and RANTES were preferentially abundant in triple-negative breast cancers (TNBCs) compared to Luminal A subtype cancers. The latter observation corresponds with the high level of TILs in the TNBC samples. IL-1β, IL-7, IL-10, and PDGFβ also exhibited a correlation between the TIF samples and matched sera. In a survival analysis, high levels of IL-5, a hallmark TH2 cytokine, in the TIF samples were associated with a worse prognosis. These findings have important implications for BC immunotherapy research.

  17. The Identification of Congeners and Aliens by Drosophila Larvae

    PubMed Central

    Del Pino, Francisco; Jara, Claudia; Pino, Luis; Medina-Muñoz, María Cristina; Alvarez, Eduardo; Godoy-Herrera, Raúl

    2015-01-01

    We investigated the role of Drosophila larva olfactory system in identification of congeners and aliens. We discuss the importance of these activities in larva navigation across substrates, and the implications for allocation of space and food among species of similar ecologies. Wild type larvae of cosmopolitan D. melanogaster and endemic D. pavani, which cohabit the same breeding sites, used species-specific volatiles to identify conspecifics and aliens moving toward larvae of their species. D. gaucha larvae, a sibling species of D. pavani that is ecologically isolated from D. melanogaster, did not respond to melanogaster odor cues. Similar to D. pavani larvae, the navigation of pavani female x gaucha male hybrids was influenced by conspecific and alien odors, whereas gaucha female x pavani male hybrid larvae exhibited behavior similar to the D. gaucha parent. The two sibling species exhibited substantial evolutionary divergence in processing the odor inputs necessary to identify conspecifics. Orco (Or83b) mutant larvae of D. melanogaster, which exhibit a loss of sense of smell, did not distinguish conspecific from alien larvae, instead moving across the substrate. Syn97CS and rut larvae of D. melanogaster, which are unable to learn but can smell, moved across the substrate as well. The Orco (Or83b), Syn97CS and rut loci are necessary to orient navigation by D. melanogaster larvae. Individuals of the Trana strain of D. melanogaster did not respond to conspecific and alien larval volatiles and therefore navigated randomly across the substrate. By contrast, larvae of the Til-Til strain used larval volatiles to orient their movement. Natural populations of D. melanogaster may exhibit differences in identification of conspecific and alien larvae. Larval locomotion was not affected by the volatiles. PMID:26313007

  18. Unraveling the Role of Surface Mucus-Binding Protein and Pili in Muco-Adhesion of Lactococcus lactis

    PubMed Central

    Duviau, Marie-Pierre; Meyrand, Mickael; Guérardel, Yann; Castelain, Mickaël; Loubière, Pascal; Chapot-Chartier, Marie-Pierre; Dague, Etienne; Mercier-Bonin, Muriel

    2013-01-01

    Adhesion of bacteria to mucus may favor their persistence within the gut and their beneficial effects to the host. Interactions between pig gastric mucin (PGM) and a natural isolate of Lactococcus lactis (TIL448) were measured at the single-cell scale and under static conditions, using atomic force microscopy (AFM). In parallel, these interactions were monitored at the bacterial population level and under shear flow. AFM experiments with a L. lactis cell-probe and a PGM-coated surface revealed a high proportion of specific adhesive events (60%) and a low level of non-adhesive ones (2%). The strain muco-adhesive properties were confirmed by the weak detachment of bacteria from the PGM-coated surface under shear flow. In AFM, rupture events were detected at short (100−200 nm) and long distances (up to 600−800 nm). AFM measurements on pili and mucus-binding protein defective mutants demonstrated the comparable role played by these two surface proteinaceous components in adhesion to PGM under static conditions. Under shear flow, a more important contribution of the mucus-binding protein than the pili one was observed. Both methods differ by the way of probing the adhesion force, i.e. negative force contact vs. sedimentation and normal-to-substratum retraction vs. tangential detachment conditions, using AFM and flow chamber, respectively. AFM blocking assays with free PGM or O-glycan fractions purified from PGM demonstrated that neutral oligosaccharides played a major role in adhesion of L. lactis TIL448 to PGM. This study dissects L. lactis muco-adhesive phenotype, in relation with the nature of the bacterial surface determinants. PMID:24260308

  19. Anti-PD-L1 treatment enhances antitumor effect of everolimus in a mouse model of renal cell carcinoma.

    PubMed

    Hirayama, Yukiyoshi; Gi, Min; Yamano, Shotaro; Tachibana, Hirokazu; Okuno, Takahiro; Tamada, Satoshi; Nakatani, Tatsuya; Wanibuchi, Hideki

    2016-12-01

    Immunotherapy based on blockade of the programmed death-1 (PD-1)/programmed death-ligand 1 (PD-L1) axis has shown promising clinical activity for renal cell carcinoma (RCC) patients; however, the most effective use of these agents in combination with conventional targeted therapy remains to be resolved. Here we evaluated the therapeutic efficacy of the combination of the mTOR inhibitor everolimus (EVE) and anti-PD-L1 using an immunocompetent mouse model of RCC. We first assessed the in vitro effect of EVE on PD-L1 expression in the human 786-O and mouse RENCA RCC cell lines and found that EVE upregulated PD-L1 expression in these RCC cell lines. We then treated RENCA tumor-bearing mice with EVE and found that PD-L1 expression was also increased in tumor cells after EVE treatment. To determine the antitumor effects of EVE alone, anti-PD-L1 alone, and EVE in combination with anti-PD-L1, we evaluated their antitumor effects on RENCA tumor-bearing mice. A significant decrease in the tumor burden was observed in the EVE alone but not in the anti-PD-L1 alone treatment group compared with the control group. Importantly, the combination of EVE with anti-PD-L1 significantly reduced tumor burden compared with the EVE alone treatment, increasing tumor infiltrating lymphocytes (TILs) and the ratio of cytotoxic CD8(+) T cells to TILs. The results of the present study demonstrated that anti-PD-L1 treatment enhanced the antitumor effect of EVE in a mouse model, supporting a direct translation of this combination strategy to the clinic for the treatment of RCC.

  20. Compensatory upregulation of PD-1, LAG-3, and CTLA-4 limits the efficacy of single-agent checkpoint blockade in metastatic ovarian cancer

    PubMed Central

    Francois, Ariel; McGray, AJ Robert; Miliotto, Anthony; Odunsi, Kunle

    2017-01-01

    ABSTRACT Tumor-associated or -infiltrating lymphocytes (TALs or TILs) co-express multiple immune inhibitory receptors that contribute to immune suppression in the ovarian tumor microenvironment (TME). Dual blockade of PD-1 along with LAG-3 or CTLA-4 has been shown to synergistically enhance T-cell effector function, resulting in a delay in murine ovarian tumor growth. However, the mechanisms underlying this synergy and the relative contribution of other inhibitory receptors to immune suppression in the ovarian TME are unknown. Here, we report that multiple immune checkpoints are expressed in TALs and TILs isolated from ovarian tumor-bearing mice. Importantly, blockade of PD-1, LAG-3, or CTLA-4 alone using genetic ablation or blocking antibodies conferred a compensatory upregulation of the other checkpoint pathways, potentiating their capacity for local T-cell suppression that, in turn, could be overcome through combinatorial blockade strategies. Whereas single-agent blockade led to tumor outgrowth in all animals, dual antibody blockade against PD-1/CTLA-4 or triple blockade against PD-1/LAG-3/CTLA-4 resulted in tumor-free survival in 20% of treated mice. In contrast, dual blockade of LAG-3 and CTLA-4 pathways using PD-1 knockout mice led to tumor-free survival in 40% of treated mice, suggesting a hierarchical ordering of checkpoint function. Durable antitumor immunity was most strongly associated with increased numbers of CD8+ T cells, the frequency of cytokine-producing effector T cells, reduced frequency of Tregs and arginine-expressing monocytic myeloid-derived suppressor cells in the peritoneal TME. These data provide a basis for combinatorial checkpoint blockade in clinical intervention for ovarian cancer. PMID:28197366

  1. Temperature controlled ionic liquid-based dispersive micro-extraction using two ligands, for determination of aluminium in scalp hair samples of Alzheimer's patients: a multivariate study.

    PubMed

    Arain, Mariam S; Arain, Salma A; Kazi, Tasneem G; Afridi, Hassan I; Ali, Jamshaid; Naeemulllah; Arain, Sadaf S; Brahman, Kapil Dev; Mughal, Moina Akhtar

    2015-02-25

    A green and sensitive temperature controlled dispersive liquid-liquid microextraction (TIL-DLLME) methodology based on the application of ionic liquid 1-butyl-3-methylimidazolium hexafluorophosphate, [C4mim][PF6], as an extractant solvent was proposed for the preconcentration of trace levels of aluminium (Al(3+)) in scalp hair samples of Alzheimer's (AD) patients, prior to analyzing by flame atomic absorption spectrometry (FAAS). The Al(3+) was complexed with 8-hydrooxyquinoline (oxine) (L1) and 3,5,7,2'-4' pentahydroxy flavone (morin) (L2) separately and then extracted by IL at temperature (50±2.0°C). Some effective factors that influence the TIL-DLLME efficiency such as pH, ligands concentrations, volume of IL, ionic strength, and incubation time were investigated and optimized by multivariate analysis. In the optimum experimental conditions, the limit of detection (3s) and enhancement factor were 0.56 μg L(-1), 0.64 μg L(-1) and 85, 73 for both ligands, respectively. The relative standard deviation (RSD) for six replicate determinations of 100 μg L(-1) Al(3+) complexed with oxine and morin were found to be 3.88% and 4.74%, respectively. The developed method was validated by the analysis of certified reference material of human hair (NCSZC81002).and applied satisfactorily to the determination of Al(3+) in acid digested scalp hair samples of AD patients and healthy controls. The resulted data shows significant higher level in scalp hair samples of AD male patients with related to referents of same age and socioeconomic status.

  2. T-lymphocyte homing: an underappreciated yet critical hurdle for successful cancer immunotherapy.

    PubMed

    Sackstein, Robert; Schatton, Tobias; Barthel, Steven R

    2017-03-27

    Advances in cancer immunotherapy have offered new hope for patients with metastatic disease. This unfolding success story has been exemplified by a growing arsenal of novel immunotherapeutics, including blocking antibodies targeting immune checkpoint pathways, cancer vaccines, and adoptive cell therapy (ACT). Nonetheless, clinical benefit remains highly variable and patient-specific, in part, because all immunotherapeutic regimens vitally hinge on the capacity of endogenous and/or adoptively transferred T-effector (Teff) cells, including chimeric antigen receptor (CAR) T cells, to home efficiently into tumor target tissue. Thus, defects intrinsic to the multi-step T-cell homing cascade have become an obvious, though significantly underappreciated contributor to immunotherapy resistance. Conspicuous have been low intralesional frequencies of tumor-infiltrating T-lymphocytes (TILs) below clinically beneficial threshold levels, and peripheral rather than deep lesional TIL infiltration. Therefore, a Teff cell 'homing deficit' may arguably represent a dominant factor responsible for ineffective immunotherapeutic outcomes, as tumors resistant to immune-targeted killing thrive in such permissive, immune-vacuous microenvironments. Fortunately, emerging data is shedding light into the diverse mechanisms of immune escape by which tumors restrict Teff cell trafficking and lesional penetrance. In this review, we scrutinize evolving knowledge on the molecular determinants of Teff cell navigation into tumors. By integrating recently described, though sporadic information of pivotal adhesive and chemokine homing signatures within the tumor microenvironment with better established paradigms of T-cell trafficking under homeostatic or infectious disease scenarios, we seek to refine currently incomplete models of Teff cell entry into tumor tissue. We further summarize how cancers thwart homing to escape immune-mediated destruction and raise awareness of the potential impact of immune

  3. Activation of aryl hydrocarbon receptor promotes invasion of clear cell renal cell carcinoma and is associated with poor prognosis and cigarette smoke.

    PubMed

    Ishida, Masaru; Mikami, Shuji; Shinojima, Toshiaki; Kosaka, Takeo; Mizuno, Ryuichi; Kikuchi, Eiji; Miyajima, Akira; Okada, Yasunori; Oya, Mototsugu

    2015-07-15

    Although exposure to environmental pollutants is one of the risk factors for renal cell carcinoma (RCC), its relationship with carcinogenesis and the progression of RCC remains unknown. The present study was designed to elucidate the role of the aryl hydrocarbon receptor (AhR), a major mediator of carcinogenesis caused by environmental pollutants, in the progression of RCC. The expression of AhR was investigated in 120 patients with RCC using immunohistochemistry, and its relationship with clinicopathological parameters and prognoses was statistically analyzed. RCC cell lines were exposed to indirubin or 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), AhR ligands, to activate the AhR pathway, or were transfected with small interfering RNA (siRNA) for AhR. The expression of the AhR target genes CYP1A1 and CYP1B1, matrix metalloproteinases (MMPs), and invasion through Matrigel(TM) were then examined. AhR was predominantly expressed in the nuclei of high-grade clear cell RCC (ccRCC) and tumor-infiltrating lymphocytes (TILs), and its expression levels in cancer cells and TILs correlated with the pathological tumor stage and histological grade. A multivariate Cox analysis revealed that the strong expression of AhR in cancer cells was a significant and independent predictor of disease-specific survival. AhR ligands up-regulated the expression of AhR and CYPs and promoted invasion by up-regulating MMPs. Furthermore, siRNA for AhR down-regulated CYPs, and inhibited cancer cell invasion together with the down-regulation of MMPs. These results suggest that AhR regulates the invasion of ccRCC and may be involved in tumor immunity. Therefore, inhibiting the activation of AhR may represent a potentially attractive therapeutic target for ccRCC patients.

  4. Importance of CD200 expression by tumor or host cells to regulation of immunotherapy in a mouse breast cancer model

    PubMed Central

    Curry, Anna; Khatri, Ismat; Kos, Olha; Zhu, Fang; Gorczynski, Reginald

    2017-01-01

    Cell-surface CD200 expression by mouse EMT6 breast tumor cells increased primary tumor growth and metastasis to the draining lymph nodes (DLN) in normal (WT) BALB/c female recipients, while lack of CD200R1 expression in a CD200R1-/- host negated this effect. Silencing CD200 expression in EMT6siCD200 tumor cells also reduced their ability to grow and metastasize in WT animals. The cellular mechanisms responsible for these effects have not been studied in detail. We report characterization of tumor infiltrating (TILs) and draining lymph node (DLN) cells in WT and CD200-/- BALB/c mice, receiving WT tumor cells, or EMT6 lacking CD200 expression (EMT6siCD200 cells). Our data show an important correlation with augmented CD8+ cytotoxic T cells and resistance to tumor growth in mice lacking exposure (on either host cells or tumor) to the immunoregulatory molecule CD200. Confirmation of the importance of such CD8+ cells came from monitoring tumor growth and characterization of the TILs and DLN cells in WT mice challenged with EMT6 and EMT6siCD200 tumors and treated with CD8 and CD4 depleting antibodies. Finally, we have assessed the mechanisms(s) whereby addition of metformin as an augmenting chemotherapeutic agent in CD200-/- animals given EMT6 tumors and treated with a previously established immunotherapy regime can increase host resistance. Our data support the hypothesis that increased autophagy in the presence of metformin increases CD8+ responses and tumor resistance, an effect attenuated by the autophagy inhibitor verteporfin. PMID:28234914

  5. Local morphologic scale: application to segmenting tumor infiltrating lymphocytes in ovarian cancer TMAs

    NASA Astrophysics Data System (ADS)

    Janowczyk, Andrew; Chandran, Sharat; Feldman, Michael; Madabhushi, Anant

    2011-03-01

    classes based on local structural properties. In this paper, we apply LMS to the specific problem of classifying regions of interest in Ovarian Cancer (OCa) histology images as either tumor or stroma. This approach is used to classify lymphocytes as either tumor infiltrating lymphocytes (TILs) or non-TILs; the presence of TILs having been identified as an important prognostic indicator for disease outcome in patients with OCa. We present preliminary results on the tumor/stroma classification of 11,000 randomly selected locations of interest, across 11 images obtained from 6 patient studies. Using a Probabilistic Boosting Tree (PBT), our supervised classifier yielded an area under the receiver operation characteristic curve (AUC) of 0.8341 +/-0.0059 over 5 runs of randomized cross validation. The average LMS computation time at every spatial location for an image patch comprising 2000 pixels with 24 particles at every location was only 18s.

  6. Position paper on the importance of psychosocial factors in cardiology: Update 2013.

    PubMed

    Ladwig, Karl-Heinz; Lederbogen, Florian; Albus, Christian; Angermann, Christiane; Borggrefe, Martin; Fischer, Denise; Fritzsche, Kurt; Haass, Markus; Jordan, Jochen; Jünger, Jana; Kindermann, Ingrid; Köllner, Volker; Kuhn, Bernhard; Scherer, Martin; Seyfarth, Melchior; Völler, Heinz; Waller, Christiane; Herrmann-Lingen, Christoph

    2014-01-01

    Hintergrund: Die rasche Weiterentwicklung der psychokardiologischen Forschung, aber auch die wachsende Verankerung psychosozialer Fragestellungen im klinischen Alltag haben die Klinische Kommission der DGK bewogen, einer Aktualisierung und Weiterentwicklung des 2008 erstmals publizierten Positionspapiers zur Bedeutung psychosozialer Faktoren in der Kardiologie zuzustimmen.Methoden: Der Kreis der Autoren wurde vergrößert, allgemeine Aspekte eingefügt und das Wissen in allen Abschnitten auf den heutigen Stand gebracht. Schwerpunkte der Empfehlungen sind die koronare Herzerkrankung, Herzrhythmusstörungen und die Herzinsuffizienz, da hier der Stand der empirischen Evidenz und des klinisches Wissens zu psychosozialen Fragestellungen am weitesten entwickelt ist. Berücksichtigt wurden bei den Empfehlungen Besonderheiten von Frauen und Männern, Unterschiede bzgl. der Lebensspanne, Einflüsse auf die kognitive Leistungsfähigkeit und die interaktive synergistische Bedeutung klassischer Risikofaktoren bei affektiver Komorbidität.Ergebnisse: Eine I-A-Empfehlung (Empfehlungsgrad I, Evidenzgrad A) wurde vergeben für die Aufforderung, psychosoziale Risikofaktoren bei der Einschätzung des KHK-Risikos zu berücksichtigen, die als unabhängige ätiologische und prognostische Risikofaktoren für das Auftreten der koronaren Herzerkrankung (KHK) und für Komplikationen im Behandlungsverlauf der KHK bedeutsam sind. Ferner für die Empfehlung, Patienten mit Herzoperationen von einem interdisziplinären Team zu betreuten, in dem die Möglichkeit besteht, auf psychosoziale Aspekte einzugehen, da bei diesen Patienten komorbide psychische Störungen wie Depressivität, Angst und posttraumatische Belastungsstörung häufig und prognostisch ungünstig sind. Eine I-B-Empfehlung wurde vergeben für die Behandlung psychosozialer Risikofaktoren mit dem Ziel einer Primärprävention der KHK, wenn das Risikomerkmal an sich Krankheitswert hat (z. B. Depression) oder die Behandlung

  7. Preface: SQM2006

    NASA Astrophysics Data System (ADS)

    Barish, Kenneth; Zhong Huang, Huan; Kapusta, Joseph; Odyniec, Grazyna; Rafelski, Johann; Whitten, Charles A., Jr.

    2006-12-01

    served as conference coordinator. She and her crew organized all the conference activities and she has been essential for the success of the conference. Staff members from UCLA, Vahe Ghazikhanian, Stephen Trentalange, Mauro Leonardo, Josephine Morrell, Friedel Adler, Tuyet-Hong Truong-Ung, Leticia Cabeza, Karin Nachtigal, Martin Simon, Dylan Thein, Lingyu Xu and Craig Reaves, and graduate students Jingguo Ma, Johan Gonzalez, Xiaoyan Lin, Priscilla Kurnadi and David Staszak, also provided essential support for the conference. We wish to thank Tony Chan, Dean of Physical Sciences at UCLA, Lawrence Berkeley National Laboratory, Brookhaven National Laboratory and the UCLA Department of Physics and Astronomy for financial support of the conference. Their sponsorship allowed many graduate students and junior physicists to attend the conference. We also thank the SQM2006 International Advisory Committee for their valuable input to the scientific programme and conference arrangement.

  8. Analysis of Contaminant Transport through the Vadose and Saturated Zones for Source Screening

    NASA Astrophysics Data System (ADS)

    Bedekar, V.; Neville, C. J.; Tonkin, M. J.

    2010-12-01

    result, the screening tool enables rapid, conservative (i.e., protective) screening-level calculations and is also ideal for providing instruction on the underlying principles. References [1] Cleary, R.W., and M.J. Ungs. 1978. Analytical models for Groundwater Pollution and Hydrology. Report 78-WR-15, Department of Civil Engineering, Princeton University, Princeton, New Jersey. [2] Neville, C.J., 2005: ATRANS: Analytical Solutions for Three Dimensional Solute Transport from a Patch Source, Version 2, S.S. Papadopulos & Associates, Inc., Waterloo, Ontario. [http://www.sspa.com/Software/atrans.shtml] [3] Ogata, A., and R.B. Banks, 1961: A Solution of the Differential Equation of Longitudinal Dispersion in Porous Media, USGS Professional Paper 411 A. [4] Wexler, E., 1992. Analytical Solutions for One-, Two, and Three-Dimensional Solute Transport in Groundwater Systems with Uniform Flow, Techniques of Water Resources Investigations of the United States Geological Survey, Chapter B-7, Book 3, Applications of Hydraulics. USGS, Washington, DC.

  9. New Experiences With Steam Injection From The MÜhlacker Field Site

    NASA Astrophysics Data System (ADS)

    Theurer, T.; Koschitzky, H.-P.; Färber, A.

    water lead to a "clog- ging" effect in the soil, displacing the steam travelling through the pore space. The use of steam with low water content was important to avoid displacement of the heat front. It also was found that conductive heat transport during breaks in the steam injec- tion process significantly warmed soil regions with low temperature, while convective heat transport brought energy to the steam front. Weak temperature gradients result- ing from a slow heating process did appear to avoid excessive saturation of the soil, thereby avoiding mobilization of the TCE. 1 Despite the low permeability of the subsurface, steam injection was found to be suc- cessful at this site. A cost of approximately 175/kg of removed chlorinated hydro- carbon through use of steam injection, compared to an estimated cost of 250/kg for cold SVE (assuming SVE would be able to remove the same amount of contaminant), was determined. Additionally, employment of steam injection has been estimated to achieve an 8.5-year time saving over the use of SVE. References: FÄRBER, A. (1997): Wärmetransport in der ungesättigten Bodenzone: Entwicklung einer ther- mischen in-situ Sanierungstechnologie. Mitteilungen des Instituts für Wasserbau, H. 96, Uni- versität Stuttgart 1997 KOSCHITZKY, H.-P., THEURER, T. (2001): Steam Injection used in unsaturated zone at german landfill. TechTrends, No. 42, Sep. 2001, United States Environmental Protection Agency (EPA), http://clu-in.org KOSCHITZKY, H.-P., THEURER, T., FÄRBER, A. (2001): Einsatz des thermischen In-situ- Sanierungsverfahrens TUBA unter schwierigen geologischen Bedingungen. Boden und Altlas- ten Symposium 2001, Erich Schmidt Verlag, 323-347 2

  10. Upscaling of lysimeter measurements to regional groundwater nitrate distribution

    NASA Astrophysics Data System (ADS)

    Klammler, Gernot; Fank, Johann; Kupfersberger, Hans; Rock, Gerhard

    2015-04-01

    generates sequences of crop rotations derived from municipal statistical data. Required retention and unsaturated hydraulic conductivity curves are derived from information out of the Austrian Soil Mapping (BMLF, 1974) by pedotransferfunctions. In summary, we present a method to quantify the temporal and spatial nitrate distribution at the aquifer scale with focus on the importance of lysimeter measurements also for modelling issues. BMLF (1974) Österreichische Bodenkartierung - Erläuterungen zur Bodenkarte 1:25.000 (Kartierungsbereich Leibnitz, Steiermark). Bundesministerium für Land- und Forstwirtschaft, Vienna, Austria Diersch HJG (2009) FEFLOW Reference Manual, DHI-WASY GmbH, Berlin Feichtinger, F. (1998). STOTRASIM - Ein Modell zur Simulation der Stickstoffdynamik in der ungesättigten Zone eines Ackerstandortes. Schriftenreihe des Bundesamtes für Wasserwirtschaft, Bd. 7, 14-41. Groenendijk, P., M. Heinen, G. Klammler, J. Fank, H. Kupfersberger, V. Pisinaras, A. Gemitzi, S. Peña-Haro, A. García-Prats, M. Pulido-Velazquez, A. Perego, M. Acutis, M. Trevisan (2014): Performance assessment of nitrate leaching models for highly vulnerable soils used in low-input farming based on lysimeter data. Sci. Tot. Environ. 499:463-480. Klammler, G., Rock, G., Fank, J. & H. Kupfersberger, H. (2011): Generating land use information to derive diffuse water and nitrate transfer as input for groundwater modelling at the aquifer scale, Proc of MODELCARE 2011 Models - Repository of Knowledge, Leipzig. Stenitzer, E. (1988). SIMWASER - Ein numerisches Modell zur Simulation des Bodenwasserhaushaltes und des Pflanzenertrages eines Standortes. Mitteilung Nr. 31, Bundesanstalt für Kulturtechnik und Bodenwasserhaushalt, A-3252 Petzenkirchen.

  11. [Transylvanian refugees and the plague in 1708-1709].

    PubMed

    Kis, D

    1993-01-01

    Owing to the overwhelming military power of the Habsburg forces Transylvanian sympathizers fled twice to Hungary during the Rákóczi uprising (1704-1711): first in 1704-1706 and then in 1707-1711. In the autumn of 1707 they numbered as much as ten thousand people, and according to the decrees of the diet at Kisvárda, they were settled down in smaller units in around Szabolcs, Szatmár, Bereg, Ung, Ugocsa and Máramaros counties. Though always short of money, the leaders of the ukprising created a system that satisfied the basic needs of these refugees. By the end of the rebellion, nevertheless, as the territory controlled by Rákóczi's armies decreased considerably, the refugees were forced to move on and on, which certainly led to a corruption of their food supplies, accommodation and hygienic conditions. The worst among all came with the plague. The author examines the effects of the epidemic and the counter-measures taken by the individual and the authorities of the uprising. Kis has consulted the main Hungarian books of that age that referred to black death (among others Anna Zay's Herbarium [1719], Samuel Köleséri's Pestis Daicae, György Komáromi Csipkés's Pestis pestise, Ferenc Pápai Páriz's Pax Corporis and A [estos betegség etc., and Máté Tsanaki's A Döghalálról, etc.), as well as many archival papers (the correspondence of Count Sándor Károlyi with his wife Krisztina Barkóczy, thos of General Bercsényi to his wife and Prince Rákóczi, and some doctors' reports, etc.). His main source, however, is Zsigmond Szaniszló's diary. Szaniszló was a former fobiró (chief-sheriff) of the Transsylvanian Torda city, an Anti-Trinitarian stronghold, and remained a leader of his people during the emigration. According to his notes, which the author has compared with the data given by the others, there were hardly any measures taken against plague in this community. Although Szaniszló gives detailed descriptions about the everyday life of the

  12. The sialotranscriptome of Antricola delacruzi female ticks is compatible with non-hematophagous behavior and an alternative source of food.

    PubMed

    Ribeiro, José Marcos C; Labruna, Marcelo B; Mans, Ben J; Maruyama, Sandra Regina; Francischetti, Ivo M B; Barizon, Gustavo Canavaci; de Miranda Santos, Isabel K F

    2012-05-01

    The hosts for Antricola delacruzi ticks are insectivorous, cave-dwelling bats on which only larvae are found. The mouthparts of nymphal and adult A. delacruzi are compatible with scavenging feeding because the hypostome is small and toothless. How a single blood meal of a larva provides energy for several molts as well as for oviposition by females is not known. Adults of A. delacruzi possibly feed upon an unknown food source in bat guano, a substrate on which nymphal and adult stages are always found. Guano produced by insectivorous bats contains twice the amount of protein and 60 times the amount of iron as beef. In addition, bacteria and chitin-rich fungi proliferate on guano. Comparative data on the transcriptome of the salivary glands of A. delacruzi is nonexistent and would help to understand the physiological adaptations of salivary glands that accompany different sources of food as well as the steps taken by the Acari toward haematophagy, believed to have evolved from scavenging dead animals. Annotation of the transcriptome of salivary glands from female instars of A. delacruzi collected on guano categorized 5.7% of the clusters of expressed genes as putative secreted proteins. They included abundantly expressed TIL-domain-containing proteins (possible anti-microbials), an abundantly expressed protein similar to a serum amyloid found in the sialotranscriptomes of Ornithodoros spp., a savignygrin, a family of mucin/peritrophin/cuticle-like proteins, anti-microbials and an HIV envelope-like glycoprotein also found in soft ticks. When comparing the transcriptome of A. delacruzi with those of blood-feeding female soft and hard ticks some notable differences were observed; they consisted of the following transcripts over- or under-represented or absent in the sialotranscriptome of A. delacruzi that may reflect its source of food: ferritin, mucins with chitin-binding domains and TIL-domain-containing proteins versus lipocalins, basic tail proteins

  13. Timescales of transport from the troposphere into the lowermost stratosphere

    NASA Astrophysics Data System (ADS)

    Boenisch, Harald; Hoor, Peter; Wernli, Heini

    2010-05-01

    The lowermost stratosphere (LMS) as part of the extratropical UTLS can be divided into dynamically and chemically distinct regions. A layer of mixed tropospheric and stratospheric tracer characteristics in the proximity of the extratropical tropopause: the extratropical tropopause transition layer (ExTL). This chemically distinct layer roughly coincides with a layer of strongly enhanced thermal stratification: the tropopause inversion layer (TIL) (Birner, 2006). The LMS above the ExTL, also named the free LMS (Bönisch et al., 2009), is less coupled to the local extratropical troposphere. Simultaneous in-situ measurements of CO2 and SF6 have been used to calculate mean transport time from the troposphere to the measurement location in the free LMS (Bönisch et al., 2009) which is on the order of months. In this study, we will use backward trajectories driven by operational ECMWF analyses wind fields to investigate the TST timescales into the LMS using the LAGRANTO scheme (Wernli and Davies, 1997). We applied a statistical data set of trajectories, which were initialized on isentropes above the 2 PVU surface up to 450K and calculated backward over 270 days (9 month) for our analysis. The results will be compared with the results from mass balance studies based on in-situ observations (Hoor et al., 2005; Bönisch et al., 2009). Furthermore, a focus is on the role of timescales of TIL formation in the LMS. Birner, T.: Fine-scale structure of the extratropical tropopause region, Journal of Geophysical Research-Atmospheres, 111, Doi 10.1029/2005jd006301, 2006. Bönisch, H., Engel, A., Curtius, J., Birner, T., and Hoor, P.: Quantifying transport into the lowermost stratosphere using simultaneous in-situ measurements of sf6 and co2, Atmospheric Chemistry and Physics, 9, 5905-5919, 2009. Hoor, P., Gurk, C., Brunner, D., Hegglin, M. I., Wernli, H., and Fischer, H.: Seasonality and extent of extratropical tst derived from in-situ co measurements during spurt, Atmospheric

  14. Successful application of spatial difference technique to electron energy-loss spectroscopy studies of Mo/SrTiO3 interfaces.

    PubMed

    Gao, M; Scheu, C; Tchernychova, E; Rühle, M

    2003-04-01

    The electron energy-loss near-edge structure (ELNES) of Mo/SrTiO3 interfaces has been studied using high spatial resolution electron energy-loss spectroscopy (EELS) in a dedicated scanning transmission electron microscope. Thin films of Mo with a thickness of 50 nm were grown on (001)-orientated SrTiO3 surfaces by molecular beam epitaxy at 600 degrees C. High-resolution transmission electron microscopy revealed that the interfaces were atomically abrupt with the (110)Mo plane parallel to the substrate surface. Ti-L2,3 ( approximately 460 eV), O-K ( approximately 530 eV), Sr-L2,3 ( approximately 1950 eV) and Mo-L2,3 ( approximately 2500 eV) absorption edges were acquired by using the Gatan Enfina parallel EELS system with a CCD detector. The interface-specific components of the ELNES were extracted by employing the spatial difference method. The interfacial Ti-L2,3 edge shifted to lower energy values and the splitting due to crystal field became less pronounced compared to bulk SrTiO3, which indicated that the Ti atoms at the interface were in a reduced oxidation state and that the symmetry of the TiO6 octahedra was disturbed. No interfacial Sr-L2,3 edge was observed, which may demonstrate that Sr atoms do not participate in the interfacial bonding. An evident interface-specific O-K edge was found, which differs from that of the bulk in both position (0.8 +/- 0.2 eV positive shift) and shape. In addition, a positive shift (0.9 +/- 0.3 eV) occurred for the interfacial Mo-L2,3, revealing an oxidized state of Mo at the interface. Our results indicated that at the interface SrTiO3 was terminated with TiO2. The validity of the spatial difference technique is discussed and examined by introducing subchannel drift intentionally.

  15. Recombinant Salmonella-based CEACAM6 and 4-1BBL vaccine enhances T-cell immunity and inhibits the development of colorectal cancer in rats: In vivo effects of vaccine containing 4-1BBL and CEACAM6.

    PubMed

    Jin, Chunhui; Liu, Yingying; Zhu, Jianhong; Xia, Tingting; Zhang, Baonan; Fei, Yajun; Ma, Bo; Ye, Jianxin; Chen, Weichang

    2015-06-01

    The present study aimed to determine the effect of recombinant Salmonella (SL3261)-based CEACAM6 and 4-1BB ligand (4-1BBL) vaccine on the development of colorectal cancer in rats and the potential immune mechanisms involved. Attenuated Salmonella typhimurium (vaccine strain)‑carrying plasmids pIRES-CEACAM6, pIRES‑4‑1BBL and pIRES-CEACAM6-4-1BBL were constructed. The rats were administered subcutaneous injections of 1,2-dimethyl-hydrazine (DMH) once a week for 18 weeks. Eight weeks after the first injection, the rats were divided into the pIRES/SL3261, pIRES-4-1BBL/SL3261, pIRES-CEACAM6/SL3261 and pIRES-CEACAM6-4-1BBL/SL3261 groups, and fed with corresponding vaccine strains. The rats were then sacrificed, the number of colon tumors were recorded, and the Dukes' stage were evaluated. CD3, CD4, CD8, CD56, FOXP3 and CEACAM6 expression in tumor tissues was determined by immunohistochemical staining. Compared with the expression levels in the pIRES/SL3261 group, similar levels of CD3+, CD8+ and CD56+ expression were identified for the pIRES-CEACAM6/SL3261 group of rats. Additionally, a comparable number of tumors was detected in the pIRES-4-1BBL/SL3261 and pIRES-CEACAM6/SL3261 groups. By contrast, a significantly fewer number of tumors, albeit with a higher density of CD3+CD8+, CD56+ and a lower density of Foxp3+ tumor-infiltrating lymphocyte (TIL) cells was detected in the pIRES-CEACAM6-4-1BBL/SL3261 group of rats. The results indicated that vaccination with recombinant attenuated Salmonella harboring the CEACAM6 and 4-1BBL gene efficiently increased the number of CD3+CD8+ TIL and NK cells, decreased the number of FOXP3 cells and inhibited the development of DMH-induced colorectal cancer in rats.

  16. Expression of programmed cell death ligand 1 is associated with poor overall survival in patients with diffuse large B-cell lymphoma

    PubMed Central

    Kiyasu, Junichi; Miyoshi, Hiroaki; Hirata, Akie; Arakawa, Fumiko; Ichikawa, Ayako; Niino, Daisuke; Sugita, Yasuo; Yufu, Yuji; Choi, Ilseung; Abe, Yasunobu; Uike, Naokuni; Nagafuji, Koji; Okamura, Takashi; Akashi, Koichi; Takayanagi, Ryoichi; Shiratsuchi, Motoaki

    2015-01-01

    Programmed cell death ligand 1 (PD-L1) is expressed on both select diffuse large B-cell lymphoma (DLBCL) tumor cells and on tumor-infiltrating nonmalignant cells. The programmed cell death 1 (PD-1)/PD-L1 pathway inhibits host antitumor responses; however, little is known about how this pathway functions in the tumor microenvironment. The aim of this study was to determine the clinicopathological impact of PD-L1+ DLBCL. We performed PD-L1/PAX5 double immunostaining in 1253 DLBCL biopsy samples and established a new definition of PD-L1+ DLBCL. We also defined the criteria for microenvironmental PD-L1+ (mPD-L1+) DLBCL (ie, PD-L1– DLBCL in which PD-L1+ nonmalignant cells are abundant in the tumor microenvironment). Of the 273 patients whose clinical information was available, quantitative analysis of PD-1+ tumor-infiltrating lymphocytes (TILs) was performed. The prevalence rates of PD-L1+ and mPD-L1+ DLBCL were 11% and 15.3%, respectively. Both PD-L1+ and mPD-L1+ DLBCL were significantly associated with non–germinal center B-cell (GCB) type and Epstein-Barr virus positivity. The number of PD-1+ TILs was significantly higher in GCB-type tumors and lower in mPD-L1– and PD-L1+ DLBCL. Patients with PD-L1+ DLBCL had inferior overall survival (OS) compared with that in patients with PD-L1– DLBCL (P = .0009). In contrast, there was no significant difference in OS between mPD-L1+ and mPD-L1– DLBCL (P = .31). The expression of PD-L1 maintained prognostic value for OS in multivariate analysis (P = .0323). This is the first report describing the clinicopathological features and outcomes of PD-L1+ DLBCL. Immunotherapy targeting the PD-1/PD-L1 pathway should be considered in this distinct DLBCL subgroup. PMID:26239088

  17. Synthesis, characterization, and reactivity of Ti(IV)-monosubstituted Keggin polyoxometalates.

    PubMed

    Kholdeeva, Oxana A; Trubitsina, Tatiana A; Maksimov, Gennadii M; Golovin, Anatolii V; Maksimovskaya, Raisa I

    2005-03-07

    Ti(IV)-monosubstituted Keggin-type polyoxometalates (Ti-POMs), mu-oxo dimer [Bu4N]8[(PTiW11O39)2O] (1), and three monomers [Bu4N]4[PTi(L)W11O39], where L = OH (2), OMe (3), and OAr (4, ArOH = 2,3,6-trimethylphenol (TMP)), have been prepared starting from mu-hydroxo dimer [Bu4N]7[(PTiW11O39)2OH] (5) or heteropolyacid H5PW11TiO40 or both. The compounds have been characterized by elemental analysis, IR, UV-vis, and multinuclear (31P, 1H, 183W) NMR. The interaction of 1 and 3-5 with H2O in MeCN produces 2. The hydrolysis constants, estimated from 31P and 1H NMR data, are 0.006 and 0.04 for 1 and 3, respectively. Studies by 31P NMR, IR, potentiometric titration, and cyclic voltammetry revealed that 1-3 and 5 afford the same protonated titanium peroxo complex [Bu4N]4[HPTi(O2)W11O39] (I) upon interaction with aqueous H2O2 in MeCN. The rates of formation of I correlate with the rates of hydrolysis of the Ti-POMs and follow the order of 5 > 1 > 3. A two-step mechanism of the reaction of Ti-POMs with H2O2, which involves hydrolysis of the Ti-L bonds to yield 2 followed by fast interaction of 2 with hydrogen peroxide producing I, is suggested. The equilibrium constant for the reaction of 2 with H2O2 to yield I and H2O, estimated using 31P NMR, is 10. The interaction of the Ti-POMs with TMP follows the trends similar to their interaction with H2O) and requires preliminary hydrolysis of the Ti-L bonds. All of the Ti-POMs catalyze the oxidation of TMP with H2O2 in MeCN to give 2,3,5-trimethyl-p-benzoquinone and 2,2',3,3',5,5'-hexamethyl-4,4'-biphenol. The product distribution is similar for all of the Ti-POMs. The catalytic activities of the Ti-POMs correlate with the rates of formation of I and follow the order of 2 > 5 > 1 > 3. The findings lay a basis for a better understanding of the nature of the reactivity of titanium in Ti-catalyzed oxidations.

  18. The promise of PD-1 inhibitors in gastro-esophageal cancers: microsatellite instability vs. PD-L1

    PubMed Central

    Jin, Zhaohui

    2016-01-01

    Preliminary clinical studies of anti-programmed cell death-1 (anti-PD-1) therapy in gastro-esophageal cancers have suggested promising single-agent activity. In patients who received prior treatment for advanced disease, pembrolizumab has been associated with a response rate of 20% in programmed cell death-1 ligand 1 (PD-L1)-positive tumors, and nivolumab with a response rate of 12% in unselected tumors. Both agents yielded a median duration of response lasting ~6–7 months. PD-L1 expression and microsatellite instability (MSI) have emerged as potential predictive markers for PD-1/PD-L1 blockade. PD-L1 expression in tumor cells and in immune cells within the tumor microenvironment has been detected in 14–24% and ~35% of patients with gastro-esophageal cancer, respectively. PD-L1 tumor cell expression appears to be more common in Epstein-Barr virus (EBV)-positive gastric cancers (GCs) and has been associated with an increased density of tumor-infiltrating lymphocytes (TIL). To date, data are too sparse to determine whether PD-L1 expression predicts efficacy of anti-PD-1 therapy in gastro-esophageal cancer, but data from other tumor types have not been consistent regarding its predictive value. MSI occurs in 10–20% of gastro-esophageal cancers and arises from deficient mismatch repair (MMR). MSI is highly correlated with non-synonymous mutation burden, as well as a dense accumulation of TILs. MSI has been associated with improved response to anti-PD-1 therapy in gastrointestinal cancers. Multiple studies are ongoing which examine therapeutic blockade of the PD-1/PD-L1 axis in unselected patients with gastro-esophageal cancer, as well as patients whose tumors express PD-L1 or exhibit MSI. These studies will clarify their activity in this disease and potentially can determine whether identify a strong predictive biomarker can be identified. Checkpoint inhibition is also being studied in combination with curative-intent chemo (radio) therapy and surgery. PMID

  19. [Implications of TCGA Network Data on 2nd Generation Immunotherapy Concepts Based on PD-L1 and PD-1 Target Structures].

    PubMed

    Peters, I; Tezval, H; Kramer, M W; Wolters, M; Grünwald, V; Kuczyk, M A; Serth, J

    2015-11-01

    The era of cytokines, given to patients with metastatic renal cell carcinoma (mRCC) as part of an unspecific immunomodulatory treatment concept, seems to have ended with the introduction of targeted therapies. However, preliminary data from studies on treatment with checkpoint inhibitors (e. g. anti-PD-1 and anti-PD-L1) may point the way to second-generation immunotherapy. The rationale of such immunomodulatory treatment is to stop or interrupt the tumour from "escaping" the body's immune defence. Thompson et al. report that increased protein expression of PD-L1 (CD274/ B7-H1) in tumour cells and tumour-infiltrating immune cells (TILs; lymphocytes and histiocytes) is associated with unfavourable clinical pathological parameters as well as poor survival. In small pilot groups of mRCC patients it was found that increased PD-L1 protein expression in tumours and TILs may be correlated with the objective response to anti-PD-1 treatment. Sometimes, however, a very wide variety of response rates was observed, which raises the question if this can be explained by individual expression levels of PD-L1 (CD 274) or PD-1 (PDCD1).Recently published data from the Cancer Genome Atlas (TCGA) Kidney Renal Clear Cell Carcinoma (KIRC) Network now provide a genome-wide data base that allows us to review or validate the molecular results obtained in clear cell renal cell carcinomas (ccRCC) to date.In this study, we analysed the TCGA KIRC mRNA expression data for PD-L1 and PD-1 for a possible association with clinical pathological parameters and the survival of 417 ccRCC patients.The mRNA expression of PD-L1 in primary nephrectomy specimens revealed no significant association with unfavourable clinical parameters. Interestingly, though, a positive correlation with patient survival was found (HR=0,59, p=0,006).These results, which partly contradict the concept applied to date, point out the necessity to ascertain the characteristics of PD-L1 and PD-1 expression at mRNA and protein

  20. The promise of PD-1 inhibitors in gastro-esophageal cancers: microsatellite instability vs. PD-L1.

    PubMed

    Jin, Zhaohui; Yoon, Harry H

    2016-10-01

    Preliminary clinical studies of anti-programmed cell death-1 (anti-PD-1) therapy in gastro-esophageal cancers have suggested promising single-agent activity. In patients who received prior treatment for advanced disease, pembrolizumab has been associated with a response rate of 20% in programmed cell death-1 ligand 1 (PD-L1)-positive tumors, and nivolumab with a response rate of 12% in unselected tumors. Both agents yielded a median duration of response lasting ~6-7 months. PD-L1 expression and microsatellite instability (MSI) have emerged as potential predictive markers for PD-1/PD-L1 blockade. PD-L1 expression in tumor cells and in immune cells within the tumor microenvironment has been detected in 14-24% and ~35% of patients with gastro-esophageal cancer, respectively. PD-L1 tumor cell expression appears to be more common in Epstein-Barr virus (EBV)-positive gastric cancers (GCs) and has been associated with an increased density of tumor-infiltrating lymphocytes (TIL). To date, data are too sparse to determine whether PD-L1 expression predicts efficacy of anti-PD-1 therapy in gastro-esophageal cancer, but data from other tumor types have not been consistent regarding its predictive value. MSI occurs in 10-20% of gastro-esophageal cancers and arises from deficient mismatch repair (MMR). MSI is highly correlated with non-synonymous mutation burden, as well as a dense accumulation of TILs. MSI has been associated with improved response to anti-PD-1 therapy in gastrointestinal cancers. Multiple studies are ongoing which examine therapeutic blockade of the PD-1/PD-L1 axis in unselected patients with gastro-esophageal cancer, as well as patients whose tumors express PD-L1 or exhibit MSI. These studies will clarify their activity in this disease and potentially can determine whether identify a strong predictive biomarker can be identified. Checkpoint inhibition is also being studied in combination with curative-intent chemo (radio) therapy and surgery.

  1. Genetic Mapping Reveals Broader Role of Vrn-H3 Gene in Root and Shoot Development beyond Heading in Barley

    PubMed Central

    Bungartz, Annemarie; Muzammil, Shumaila; P. Afsharyan, Nazanin; Léon, Jens; Naz, Ali Ahmad

    2016-01-01

    The aim of the present study was to dissect the genetic inheritance and interplay of root, shoot and heading attributes for a better understanding of these traits in crop production. For this, we utilized quantitative trait loci (QTL) and candidate gene analysis approach using a second filial (F2) population originated from a cross between spring cultivar Cheri and wild barley accession ICB181160. The F2 population comprising 182 plants was phenotyped for root dry weight (RDW), root volume (RV), root length (RL) and shoot dry weight (SDW), tiller number per plant (TIL) and days to heading (HEA). In parallel, this population was genotyped using polymerase chain reaction (PCR) based cleaved amplified polymorphic sequence (CAPS) markers distributed across the whole genome. Marker by trait analysis revealed 16 QTL for root and shoot traits localized on chromosomes 1H, 3H, 4H, 5H and 7H. The strongest and a common QTL effect for root, shoot and heading traits was identified on chromosome 7H at the putative region of Vrn-H3 gene. Later, we have established PCR based gene specific marker HvVrnH3 revealing polymorphism for early heading Vrn-H3 allele in Cheri and late heading allele vrn-H3 in ICB181160. Genotyping of these alleles revealed a clear co-segregation of early heading Vrn-H3 allele with lower root and shoot attributes, while late heading vrn-H3 allele with more TIL and higher root biomass suggesting a primary insight on the function of Vrn-H3 gene beyond flowering. Genetic interactions of vernalization genes Vrn-H3 with Vrn-H2 and Vrn-H1 also suggested the major role of Vrn-H3 alleles in determining root and shoot trait variations in barley. We believe, these data provide an opportunity for further research to test a precise significance of early heading on yield components and root associated sustainability in crops like barley and wheat. PMID:27442506

  2. Dollar Summary of Federal Supply Classification and Service Category by Company. Part 7. (1005-2845)

    DTIC Science & Technology

    1989-01-01

    0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 wi 0000 000000000000000001 - U In ti-I Ut IA U tilE Ut Ut tD Ut Ut Ut U Ut Ut Ut Qt ut IA Q z )- l 1 .,4 -4 .. 4...O O O-4 4)ou o L L.0h.4 - 4 -44 𔃾 - 4 - -4 -P4 -4 -4 L. td -4 t4 -4 -4 -4 -4 L. 1 Ix 1tt 0 0 0 0 0 0 0 0 0 ’D 0 E o 0 o 0 0 0m 0 W 4 0 (0 I I 4- %- 4...m )0 z 0 0 I 1 I ) 4- 4- I.- %- 0 D C %- =---4 L1 I ZI C 4 0 >, . 4 N 0 ., 4 0 0 1 H - I =)I C 44 L - -- 4- M 4 0 4-L - 4 3 ex I o 0 4 0 td . go m0 4

  3. In vitro stimulation of stage-specific deoxyribonucleic acid synthesis in rat seminiferous tubule segments by interleukin-1. alpha

    SciTech Connect

    Parvinen, M.; Soeder, O.M.; Mali, P.; Froeysa, B.R.; Ritzen, E.M. )

    1991-09-01

    Levels of rat testicular interleukin-1-like factor (tIL-1) have been shown to correlate with DNA synthetic activity during the cycle of the rat seminiferous epithelium, suggesting its role as a spermatogonial or meiotic growth factor. To explore this further, a new in vitro model system was developed. Rat seminiferous tubule segments from stages I, V, VIIa, and VIII-IX of the cycle were isolated by transillumination-assisted microdissection, cultured in chemically defined serum-free medium supplemented with human recombinant IL-1 {alpha}, and labeled with (3H)thymidine. During incubation, spontaneous progression of spermatogenesis was noted. Inactive stage VIIa tubule segments differentiated to stage VIII and initiated DNA synthesis, and concomitantly started to secrete IL-1-like factor. DNA synthesis of stages VIII-IX ceased through differentiation of spermatocytes to leptotene-zygotene (stages XII-XIII of the cycle). IL-1 {alpha} stimulated DNA synthesis significantly in spermatogonia of stage I. Meiotic DNA synthesis at stage VIIa was stimulated (48 h/34 C) and maintained at stages VIII-IX (48 h/34 C). IL-1 {alpha} seems to act as a regulator of spermatogenic DNA synthesis in both mitotic and meiotic phases. It has mainly stimulating and maintaining effects, but it may also be inhibitory under certain conditions.

  4. Reasoning about real-time systems with temporal interval logic constraints on multi-state automata

    NASA Technical Reports Server (NTRS)

    Gabrielian, Armen

    1991-01-01

    Models of real-time systems using a single paradigm often turn out to be inadequate, whether the paradigm is based on states, rules, event sequences, or logic. A model-based approach to reasoning about real-time systems is presented in which a temporal interval logic called TIL is employed to define constraints on a new type of high level automata. The combination, called hierarchical multi-state (HMS) machines, can be used to model formally a real-time system, a dynamic set of requirements, the environment, heuristic knowledge about planning-related problem solving, and the computational states of the reasoning mechanism. In this framework, mathematical techniques were developed for: (1) proving the correctness of a representation; (2) planning of concurrent tasks to achieve goals; and (3) scheduling of plans to satisfy complex temporal constraints. HMS machines allow reasoning about a real-time system from a model of how truth arises instead of merely depending of what is true in a system.

  5. [Treatment of renal anemia in hemodialysis patients in General Hospital Bjelovar from 2007 to 2010].

    PubMed

    Lovcić, Vesna; Vujić, Jovana; Basić-Jukić, Nikolina; Kurtović, Ivana; Janković, Renata Ivanac; Lovcić, Petra; Dzapo, Marko

    2011-10-01

    Renal anemia is complication of chronic kidney disease. It is caused by crythropoietin deficency and is associated with adverse outcomes in CKD patients. Renal anemia should be treated with erythropoesis-stimulating agents (ESAs), supplementary iron, adequate dialysis, and if necessary with red blood cells transfusions. The main problem of treatment is how to determine target hemoglobin value and keep it within the constant range. Current guidelines recommend target hemoglobin level 110 - 120 g/l, but optimal value need to be adjusted for every patient individualy keeping in mind primary kidney disease, age, gender and comorbidities. In Department of Dialysis of General Hospital Bjelovar we carried out a retrospective study about treating renal anemia in 67 patients on chronic hemodialysis program during 36 months in a period from 2007. til 2010. We monitored hemoglobin, feritin, saturation of transferin (TSAT), dose of LSE, number of change in dosage and number of transfusion. Mean hemoglobin level was 107.8 g/l, feritin level 196.8 mcg/l, TSAT 24.16%, weekly dose of ESAs 5951.9 IU. in 53.7% patiens dose was changed 11 - 20 times during that period, and 34% of patiens was treated with at least 1 dose of transfusion of red blood cells. We conclude that better iron supplementation and moderately higher doses of FSAs correlate with higher hemoglobin value, and hemoglobin variations is still big problem in renal anemia treatment.

  6. Whole-Transcriptome Selection and Evaluation of Internal Reference Genes for Expression Analysis in Protocorm Development of Dendrobium officinale Kimura et Migo

    PubMed Central

    An, Hongqiang; Zhu, Qiankun; Pei, Wei; Fan, Jing; Liang, Yi; Cui, Yihui; Lv, Nan; Wang, Wanjun

    2016-01-01

    Dendrobium officinale Kimu et Migo has increased many researchers’ interest for its high medical and horticultural values and the molecular mechanism of its protocorm development remains unclear. In this study, 19 genes from 26 most stably expressed genes in whole transcriptome of protocorms and 5 housekeeping genes were used as candidate reference genes and screened with 4 application softwares (geNorm, NormFinder, BestKeeper and RefFinder). The results showed that a few reference genes could effectively normalize expression level of specific genes in protocorm development and the optimal top 2 reference genes were ASS and APH1L. Meanwhile, validation of GNOM, AP2 and temperature induced gene (TIL) for normalization demonstrates the usefulness of the validated candidate reference genes. The expression profiles of these genes varied under protocorms and temperature stress according to the stablest and unstablest reference genes, which proved the importance of the choice of appropriate reference genes. The first systematic evaluation of stably expressed genes will be very useful in the future analysis of specific genes expression in D. officinale. PMID:27814359

  7. Primitive robotic procedures: automotions for medical liquids in 12th century Asia minor.

    PubMed

    Penbegul, Necmettin; Atar, Murat; Kendirci, Muammer; Bozkurt, Yasar; Hatipoglu, Namık Kemal; Verit, Ayhan; Kadıoglu, Ates

    2014-12-30

    In recent years, day by day, robotic surgery applications have increase their role in our medical life. In this article, we reported the discovery of the first primitive robotic applications as automatic machines for the sensitive calculation of liquids such as blood in the literature. Al-Jazari who wrote the book "Elcâmi 'Beyne'l - 'ilm ve'l - 'amel en-nâfi 'fi es-sınaâ 'ti'l - hiyel", lived in Anatolian territory between 1136 and 1206. In this book that was written in the twelfth century, Al-Jazari described nearly fifty graphics of robotic machines and six of them that were designed for medical purposes. We found that some of the robots mentioned in this book are related to medical applications. This book reviews approximately 50 devices, including water clocks, candle clocks, ewers, various automata used for amusement in drink assemblies, automata used for ablution, blood collection tanks, fountains, music devices, devices for water lifting, locks, a protractor, a boat-shaped water clock, and the gate of Diyarbakir City in south-east of Turkey, actually in northern Mesopotamia. We found that automata used for ablution and blood collection tanks were related with medical applications; therefore, we will describe these robots.

  8. Evolution and classification of acapulcoites and lodranites from a chemical point of view

    NASA Astrophysics Data System (ADS)

    Patzer, Andrea; Hill, Dolores H.; Boynton, William V.

    2004-01-01

    We examined 15 bulk samples of the acapulcoite-lodranite clan for their major, minor, and trace element concentrations using INAA techniques. Among the analyzed meteorites are 2 new acapulcoites (Dhofar [Dho] 290, Thiel Mountains [TIL] 99002) as well as an additional acapulcoite that has been described previously only in very brief form (Graves Nunataks [GRA] 98028). The petrographic attributes of these 3 samples are addressed thoroughly. We also include petrographic information on 2 acapulcoites from Africa: Northwest Africa (NWA) 725 and NWA 1058. In general, our study strongly supports the widely accepted idea that acapulcoites and lodranites evolved through partial melting and melt migration of metal/sulfide phases and plagioclase. Furthermore, we concur with previous researchers that the original bimodal classification scheme for acapulcoites and lodranites proves to be too simple. Based on our data set, we introduce an alternative, extended scheme. With respect to their elemental distribution patterns, we distinguish 5 subtypes comprising primitive, typical, transitional, and enriched acapulcoites on one hand and lodranites on the other. The chemical distinction between the primitive, typical, and transitional acapulcoites is rather subtle and gradual. It stands in contrast to the clear modifications observed for the signatures of the enriched acapulcoites and the lodranites. The definition of subcategories basically reflects the concentrations of 2 key elements: K and Se. We note, however, that the assignment of subgroups may not be exclusively inferred from elemental abundances but should also consider additional petrographic information.

  9. Multispectral imaging and image processing

    NASA Astrophysics Data System (ADS)

    Klein, Julie

    2014-02-01

    The color accuracy of conventional RGB cameras is not sufficient for many color-critical applications. One of these applications, namely the measurement of color defects in yarns, is why Prof. Til Aach and the Institute of Image Processing and Computer Vision (RWTH Aachen University, Germany) started off with multispectral imaging. The first acquisition device was a camera using a monochrome sensor and seven bandpass color filters positioned sequentially in front of it. The camera allowed sampling the visible wavelength range more accurately and reconstructing the spectra for each acquired image position. An overview will be given over several optical and imaging aspects of the multispectral camera that have been investigated. For instance, optical aberrations caused by filters and camera lens deteriorate the quality of captured multispectral images. The different aberrations were analyzed thoroughly and compensated based on models for the optical elements and the imaging chain by utilizing image processing. With this compensation, geometrical distortions disappear and sharpness is enhanced, without reducing the color accuracy of multispectral images. Strong foundations in multispectral imaging were laid and a fruitful cooperation was initiated with Prof. Bernhard Hill. Current research topics like stereo multispectral imaging and goniometric multispectral measure- ments that are further explored with his expertise will also be presented in this work.

  10. Bone formation in vivo induced by Cbfa1-carrying adenoviral vectors released from a biodegradable porous β-tricalcium phosphate (β-TCP) material

    NASA Astrophysics Data System (ADS)

    Uemura, Toshimasa; Kojima, Hiroko

    2011-06-01

    Overexpression of Cbfa1 (a transcription factor indispensable for osteoblastic differentiation) is expected to induce the formation of bone directly and indirectly in vivo by accelerating osteoblastic differentiation. Adenoviral vectors carrying the cDNA of Cbfa1/til-1(Adv-Cbf1) were allowed to be adsorbed onto porous blocks of β-tricalcium phosphate (β-TCP), a biodegradable ceramic, which were then implanted subcutaneously and orthotopically into bone defects. The adenoviral vectors were released sustainingly by biodegradation, providing long-term expression of the genes. Results of the subcutaneous implantation of Adv-Cbfa1-adsorbed β-TCP/osteoprogenitor cells suggest that a larger amount of bone formed in the pores of the implant than in the control material. Regarding orthotopic implantation into bone defects, the released Adv-Cbfa1 accelerated regeneration in the cortical bone, whereas it induced bone resorption in the marrow cavity. A safer gene transfer using a smaller amount of the vector was achieved using biodegradable porous β-TCP as a carrier.

  11. An insight into the salivary transcriptome and proteome of the soft tick and vector of epizootic bovine abortion, Ornithodoros coriaceus

    PubMed Central

    Francischetti, Ivo M. B.; Meng, Zhaojing; Mans, Ben J.; Gudderra, Nanda; Hall, Mark; Veenstra, Timothy D.; Pham, Van M.; Kotsyfakis, Michail; Ribeiro, José M. C.

    2008-01-01

    The salivary glands of blood sucking arthropods contain a redundant ‘magic potion’ that counteracts their vertebrate host’s hemostasis, inflammation, and immunity. We here describe the salivary transcriptome and proteomics (sialome) of the soft tick Ornithodoros coriaceus. The resulting analysis helps to consolidate the classification of common proteins found in both soft and hard ticks, such as the lipocalins, Kunitz, cystatin, basic tail, hebraein, defensin, TIL domain, metalloprotease, 5′-nucleotidase/apyrase, and phospholipase families, and also to identify protein families uniquely found in the Argasidae, such as the adrenomedullin/CGRP peptides, 7DB, 7 kDa, and the RGD containing single Kunitz proteins. Additionally, we found a protein belonging to the cytotoxin protein family that has so far only been identified in hard ticks. Three other unique families common only to the Ornithodoros genus were discovered. Edman degradation, 2D and 1D PAGE of salivary gland homogenates followed by tryptic digestion and HPLC MS/MS of results confirms the presence of several proteins. These results indicate that each genus of hematophagous arthropods studied to date evolved unique protein families that assist blood feeding, thus characterizing potentially new pharmacologically active components or antimicrobial agents. PMID:18725333

  12. Size scales over which ordinary chondrites and their parent asteroids are homogeneous in oxidation state and oxygen-isotopic composition

    NASA Astrophysics Data System (ADS)

    Rubin, Alan E.; Ziegler, Karen; Young, Edward D.

    2008-02-01

    Literature data demonstrate that on a global, asteroid-wide scale (plausibly on the order of 100 km), ordinary chondrites (OC) have heterogeneous oxidation states and O-isotopic compositions (represented, respectively, by the mean olivine Fa and bulk Δ 17O compositions of equilibrated samples). Samples analyzed here include: (a) two H5 chondrite Antarctic finds (ALHA79046 and TIL 82415) that have the same cosmic-ray exposure age (7.6 Ma) and were probably within ˜1 km of each other when they were excavated from the H-chondrite parent body, (b) different individual stones from the Holbrook L/LL6 fall that were probably within ˜1 m of each other when their parent meteoroid penetrated the Earth's atmosphere, and (c) drill cores from a large slab of the Estacado H6 find located within a few tens of centimeters of each other. Our results indicate that OC are heterogeneous in their bulk oxidation state and O-isotopic composition on 100-km-size scales, but homogeneous on meter-, decimeter- and centimeter-size scales. (On kilometer size scales, oxidation state is heterogeneous, but O isotopes appear to be homogeneous.) The asteroid-wide heterogeneity in oxidation state and O-isotopic composition was inherited from the solar nebula. The homogeneity on small size scales was probably caused in part by fluid-assisted metamorphism and mainly by impact-gardening processes (which are most effective at mixing target materials on scales of ⩽1 m).

  13. Targeting solid tumors via T cell receptor complementarity-determining region 3delta in an engineered antibody.

    PubMed

    Wang, Zhun; Zhang, Tie; Hu, Hongbo; Zhang, Huiyuan; Yang, Zhi; Cui, Lianxian; He, Wei

    2008-12-18

    Human Vdelta2 gammadelta T lymphocytes killed multiple solid tumors, even displaying comparable therapeutic efficacy with anti-tumor chemical-cis-platinum in an adoptive experiment in both nude and SCID murine model shown in present study. We previously found that T cell receptor (TCR) gammadelta recognize tumors via complementarity-determining region 3 (CDR3), briefly named as CDR3delta. Based on characteristics of specific binding of CDR3delta to tumor targets, we developed a novel tumor-targeting antibody, whose CDR3 in heavy chain is replaced by CDR3delta sequence derived from human ovarian carcinoma (OEC) infiltrating gammadelta T cells (gammadeltaTILs). This CDR3delta-grafted antibody OT3 exhibited specific binding activities to OEC line SKOV3 both in vitro and in vivo, which included specific binding to several tumor cell lines, interacting with heat shock protein (HSP) 60 and triggering ADCC against tumors in vitro, as well as displaying tumor imaging by radioisotope 99mTc-labeled antibody OT3 in vivo. Moreover, immunotoxin OT3-DT, CDR3delta-grafted antibody OT3 chemically conjugated with diphtheria toxin (DT) showed the anti-tumor effect on the growth of several solid tumors including OEC, cervix adenocarcinoma, hepatocellular carcinoma, and rectum adenocarcinoma to various extents in nude mice. Therefore, we have found and confirmed a novel therapeutic strategy for targeting solid tumors, making use of immune recognition characteristics of gammadelta T cells.

  14. [Edge effect of the plant community structure on land-bridge islands in the Thousand Island Lake].

    PubMed

    Su, Xiao-Fei; Yuan, Jin-Feng; Hu, Guang; Xu, Gao-Fu; Yu, Ming-Jian

    2014-01-01

    The research was conducted on 29 land-bridge islands in the Thousand Island Lake (TIL), where long-term monitoring plots were set up during 2009-2010. The community attributes including species richness, Shannon index, plant mean height, plant mean diameter at breast height (DBH) and plant density along the edge-interior gradient from edge to interior forest were calculated to investigate the edge effect. The results showed that the species richness and Shannon index were affected through the whole gradient (larger than 50 m), while the range of edge effect was 20-30 m on mean plant height, and 10 m on plant density and mean DBH. Community attributes differed significantly among the edge gradients. The species richness and Shannon index peaked at the intermediate edge gradient. Plant density decreased and plant mean height increased along the edge to interior gradient. All five community attributes were significantly associated with the edge gradient, also different functional groups, evergreen or deciduous species, trees or shrubs, shade tolerant or shade intolerant species, were differentially influenced by the edge effect. It was demonstrated the influence of edge effect on the fragmented forest community varied with community attributes and functional groups.

  15. Adoptive immunotherapy for hematological malignancies using T cells gene-modified to express tumor antigen-specific receptors.

    PubMed

    Fujiwara, Hiroshi

    2014-12-15

    Accumulating clinical evidence suggests that adoptive T-cell immunotherapy could be a promising option for control of cancer; evident examples include the graft-vs-leukemia effect mediated by donor lymphocyte infusion (DLI) and therapeutic infusion of ex vivo-expanded tumor-infiltrating lymphocytes (TIL) for melanoma. Currently, along with advances in synthetic immunology, gene-modified T cells retargeted to defined tumor antigens have been introduced as "cellular drugs". As the functional properties of the adoptive immune response mediated by T lymphocytes are decisively regulated by their T-cell receptors (TCRs), transfer of genes encoding target antigen-specific receptors should enable polyclonal T cells to be uniformly redirected toward cancer cells. Clinically, anticancer adoptive immunotherapy using genetically engineered T cells has an impressive track record. Notable examples include the dramatic benefit of chimeric antigen receptor (CAR) gene-modified T cells redirected towards CD19 in patients with B-cell malignancy, and the encouraging results obtained with TCR gene-modified T cells redirected towards NY-ESO-1, a cancer-testis antigen, in patients with advanced melanoma and synovial cell sarcoma. This article overviews the current status of this treatment option, and discusses challenging issues that still restrain the full effectiveness of this strategy, especially in the context of hematological malignancy.

  16. Organization of cytoskeletal elements and organelles preceding growth cone emergence from an identified neuron in situ

    PubMed Central

    1989-01-01

    The purpose of this study was to investigate the arrangement of cytoskeletal elements and organelles in an identified neuron in situ at the site of emergence of its growth cone just before and concurrent with the onset of axonogenesis. The Ti1 pioneer neurons are the first pair of afferent neurons to differentiate in embryonic grasshopper limbs. They arise at the distal tip of the limb bud epithelium, the daughter cells of a single precursor cell, the Pioneer Mother Cell (PMC). Using immunohistochemical markers, we characterized the organization of microtubules, centrosomes, Golgi apparatus, midbody, actin filaments, and chromatin from mitosis in the PMC through axonogenesis in the Tils. Just before and concurrent with the onset of axonogenesis, a characteristic arrangement of tubulin, actin filaments, and Golgi apparatus is localized at the proximal pole of the proximal pioneer neuron. The growth cone of the proximal cell stereotypically arises from this site. Although the distal cell's axon generally grows proximally, occasionally it arises from its distal pole; in such limbs, the axons from the sister cells extend from mirror symmetric locations on their somata. In the presence of cytochalasin D, the PMC undergoes nuclear division but not cytokinesis and although other neuronal phenotypes are expressed, axongenesis is inhibited. Our data suggest that intrinsic information determines the site of growth cone emergence of an identified neuron in situ. PMID:2654140

  17. The “Vampirome”: Transcriptome and proteome analysis of the principal and accessory submaxillary glands of the vampire bat Desmodus rotundus, a vector of human rabies

    PubMed Central

    Francischetti, Ivo M. B.; Assumpção, Teresa C. F.; Ma, Dongying; Li, Yuan; Vicente, Eliane C.; Uieda, Wilson; Ribeiro, José M.C.

    2013-01-01

    Vampire bats are notorious for being the sole mammals that strictly feed on fresh blood for their survival. While their saliva has been historically associated with anticoagulants, only one antihemostatic (plasminogen activator) has been molecularly and functionally characterized. Here, RNAs from both principal submandibular and accessory glands of Desmodus rotundus were extracted, and ~ 200 million reads were sequenced by Illumina. The principal gland was enriched with plasminogen activators with fibrinolytic properties, members of lipocalin and secretoglobin families, which bind prohemostatic prostaglandins, and endonucleases, which cleave neutrophil-derived procoagulant NETs. Anticoagulant (tissue factor pathway inhibitor, TFPI), vasodilators (PACAP and C-natriuretic peptide), and metalloproteases (ADAMTS-1) were also abundantly expressed. Members of the TSG-6 (anti-inflammatory), antigen 5/CRISP, and CCL28-like (antimicrobial) protein families were also sequenced. Apyrases (which remove platelet agonist ADP), phosphatases (which degrade procoagulant polyphosphates), and sphingomyelinase were found at lower transcriptional levels. Accessory glands were enriched with antimicrobials (lysozyme, defensin, lactotransferrin) and protease inhibitors (TIL-domain, cystatin, Kazal). Mucins, heme-oxygenase, and IgG chains were present in both glands. Proteome analysis by nano LC-MS/MS confirmed that several transcripts are expressed in the glands. The database presented herein is accessible online at http://exon.niaid.nih.gov/transcriptome/D_rotundus/Supplemental-web.xlsx. These results reveal that bat saliva emerges as a novel source of modulators of vascular biology. PMID:23411029

  18. Micromechanics of Drosophila Audition

    NASA Astrophysics Data System (ADS)

    Göpfert, M. C.; Robert, D.

    2003-02-01

    An analysis is presented of the auditory micromechanics of the fruit fly Drosophila melanogaster. In this animal, the distal part of the antenna constitutes a resonantly tuned sound receiver, the vibrations of which are transduced by a chordotonal sense organ in the antenna's base. Analyzing the mechanical behavior of the antennal receiver by means of microscanning laser Doppler vibrometry, we show that the auditory system of wild-type flies exhibits a hardening stiffness nonlinearity and spontaneously generates oscillations in the absence of external stimuli. According to the deprivation of these mechanical properties in mechanosensory mutants, the receiver's nonlinearity and oscillation activity are introduced by chordotonal auditory neurons. Requiring the mechanoreceptor-specific extracellular linker protein No-mechanoreceptor-potential-A (NompA), NompC mechanosensory transduction channels, Beethoven (Btv), and Touch-insensitive-larva-B (TilB), nonlinearity and oscillation activity of the fly's antennal receiver depend on prominent components of the auditory transduction machinery and seem to originate from motility of auditory receptor cilia.

  19. Yakir Aharonov: From A to B

    NASA Astrophysics Data System (ADS)

    Pines, A.

    Twenty years ago, I was accorded the privilege and pleasure of presenting the after banquet speech—reprinted below—at a conference celebrating the sixtieth year of my dear friend Yakir Aharonov. Does what I said then still hold today? You bet it does …the now adult Yakir has maintained the same burning curiosity, fiery enthusiasm and explosive genius for physics that characterized the life of the adolescent sixty year old. There may be no free lunch, but Yakir is living proof that there is free will, and he has exercised his free will to make ever more creative contributions to quantum physics, future and present. My admiration of Yakir as a scientist and mensch has only grown over the years. Sorry I can't be with you all at Chapman to participate in this marvelous eightieth year celebration. Dear Yakir, Ditsa and I wish you, Nilli and your family many more joyous years of health, science and friendship…' til 120.

  20. Location and Density of Immune Cells in Precursor Lesions and Cervical Cancer.

    PubMed

    Bedoya, Astrid M; Jaramillo, Roberto; Baena, Armando; Castaño, Jorge; Olaya, Natalia; Zea, Arnold H; Herrero, Rolando; Sanchez, Gloria I

    2013-04-01

    Only a small proportion of women infected with Human Papillomavirus (HPV) develop cervical cancer. Host immune response seems to play a role eliminating the viral infection and preventing progression to cancer. Characterization of tumor infiltrating lymphocytes (TILs) in cervical pre-neoplastic lesions and cervical cancer may be helpful to understand the mechanisms that mediate this protection. The aim of this study was to determine if there are differences in the localization and density (cells/mm(2)) of CD8+ T-cells, CD4+ T-cells and Tregs (CD25 + Foxp3+) in cervical pre-neoplastic lesions and cervical cancer. Immunohistochemical analysis of sections of 96 (26 CIN1, 21 CIN2, 25 CIN3, and 24 SCC) samples revealed that regardless of CIN grades, CD8+ T-cells are more abundant than CD4+, CD25+ and Foxp3+ cells in both the stroma and epithelium. There was a higher density of CD8+ cells in the stroma of cervical cancer compared to CIN3 (OR = 4.20, 95% CI 1.2-15), CIN2 (OR = 7.86, 95% CI 1.7-36.4) and CIN1 (OR = 4.25, 95% CI 1.1-17). Studies evaluating whether these cells are recruited before or after cancer progression will be helpful to understand the role of these cells in the natural history of HPV-induced lesions.

  1. Popocatepetl Erupts

    NASA Technical Reports Server (NTRS)

    2002-01-01

    The Popocatepetl Volcano, almost 30 miles south of Mexico City, erupted yesterday (December 18, 2000) in what authorities are calling its most spectacular eruption since 800 A.D. This morning, Popocatepetl (pronounced poh-poh-kah-TEH-peh-til) continued spewing red-hot rocks as well as a column of smoke and ash about 2.5 miles high into the atmosphere. This true-color image of the volcano was acquired today by the Sea-viewing Wide Field-of-view Sensor (SeaWiFS) flying aboard the OrbView-2 satellite. In this image, Popocatepetl's plume (greyish pixels) can be seen blowing southward, away from Mexico City. There is a large cloud bank (bright white pixels) just to the east of the volcanic plume. Although Popocatepetl has been active since 1994-when it awoke from a 70-year slumber-this most recent eruption is most concerning to the greater Mexico City region's 20 million residents. The volcano demonstrated what it can do in 800 A.D. when it belched forth enough lava to fill many of the valleys in the surrounding region. Earlier, scientists warned the citizens of Mexico that there is a dome of lava at the base of the volcano that is causing pressure to build inside. They are concerned that, if it continues to build unabated, this pressure could cause even larger eruptions in the future. Image provided by the SeaWiFS Project, NASA/Goddard Space Flight Center, and ORBIMAGE

  2. PD-1 and its ligands are important immune checkpoints in cancer.

    PubMed

    Dong, Yinan; Sun, Qian; Zhang, Xinwei

    2017-01-10

    Checkpoint programmed death-1 (PD-1)/programmed cell death ligands (PD-Ls) have been identified as negative immunoregulatory molecules that promote immune evasion of tumor cells. The interaction of PD-1 and PD-Ls inhibits the function of T cells and tumor-infiltrating lymphocytes (TIL) while increasing the function of immunosuppressive regulatory T cells (Tregs). This condition causes the tumor cells to evade immune response. Thus, the blockade of PD-1/PD-L1 enhances anti-tumor immunity by reducing the number and/or the suppressive activity of Tregs and by restoring the activity of effector T cells. Furthermore, some monoclonal antibodies blockading PD-1/PD-Ls axis have achieved good effect and received Food and Drug Administration approval. The role of PD-1/PD-Ls in tumors has been well studied, but little is known on the mechanism by which PD-1 blocks T-cell activation. In this study, we provide a brief overview on the discovery and regulatory mechanism of PD-1 and PD-L1 dysregulation in tumors, as well as the function and signaling pathway of PD-1 and its ligands; their roles in tumor evasion and clinical treatment were also studied.

  3. Tumor-Induced CD8+ T-Cell Dysfunction in Lung Cancer Patients

    PubMed Central

    Prado-Garcia, Heriberto; Romero-Garcia, Susana; Aguilar-Cazares, Dolores; Meneses-Flores, Manuel; Lopez-Gonzalez, Jose Sullivan

    2012-01-01

    Lung cancer is the leading cause of cancer deaths worldwide and one of the most common types of cancers. The limited success of chemotherapy and radiotherapy regimes have highlighted the need to develop new therapies like antitumor immunotherapy. CD8+ T-cells represent a major arm of the cell-mediated anti-tumor response and a promising target for developing T-cell-based immunotherapies against lung cancer. Lung tumors, however, have been considered to possess poor immunogenicity; even so, lung tumor-specific CD8+ T-cell clones can be established that possess cytotoxicity against autologous tumor cells. This paper will focus on the alterations induced in CD8+ T-cells by lung cancer. Although memory CD8+ T-cells infiltrate lung tumors, in both tumor-infiltrating lymphocytes (TILs) and malignant pleural effusions, these cells are dysfunctional and the effector subset is reduced. We propose that chronic presence of lung tumors induces dysfunctions in CD8+ T-cells and sensitizes them to activation-induced cell death, which may be associated with the poor clinical responses observed in immunotherapeutic trials. Getting a deeper knowledge of the evasion mechanisms lung cancer induce in CD8+ T-cells should lead to further understanding of lung cancer biology, overcome tumor evasion mechanisms, and design improved immunotherapeutic treatments for lung cancer. PMID:23118782

  4. A pictorial presentation of 3.0 Chicago Classification for esophageal motility disorders.

    PubMed

    Herbella, Fernando Augusto; Armijo, Priscila Rodrigues; Patti, Marco Giuseppe

    2016-01-01

    High resolution manometry changed several esophageal motility paradigms. The 3.0 Chicago Classification defined manometric criteria for named esophageal motility disorders. We present a pictorial atlas of motility disorders. Achalasia types, esophagogastric junction obstruction, absent contractility, distal esophageal spasm, hypercontractile esophagus (jackhammer), ineffective esophageal motility, and fragmented peristalsis are depicted with high-resolution manometry plots. RESUMO A manometria de alta resolução mudou vários paradigmas da motilidade digestiva. A Classificação de Chicago, na versão 3.0, definiu critérios manométricos para as doenças da motilidade esofagiana. O presente artigo é um atlas das dismotilidades descritas. Tipos de acalásia, obstrução ao nível da junção esofagogástrica, contrações ausentes, espasmo esofagiano distal, esôfago hipercontrátil, motilidade esofagiana ineficaz e peristalse fragmentada são mostradas em traçados de manometria de alta resolução.

  5. Nanoparticle size and combined toxicity of TiO2 and DSLS (surfactant) contribute to lysosomal responses in digestive cells of mussels exposed to TiO2 nanoparticles.

    PubMed

    Jimeno-Romero, A; Oron, M; Cajaraville, M P; Soto, M; Marigómez, I

    2016-10-01

    The aim of this investigation was to understand the bioaccumulation, cell and tissue distribution and biological effects of disodium laureth sulfosuccinate (DSLS)-stabilised TiO2 nanoparticles (NPs) in marine mussels, Mytilus galloprovincialis. Mussels were exposed in vivo to 0.1, 1 and 10 mg Ti/L either as TiO2 NPs (60 and 180 nm) or bulk TiO2, as well as to DSLS alone. A significant Ti accumulation was observed in mussels exposed to TiO2 NPs, which were localised in endosomes, lysosomes and residual bodies of digestive cells, and in the lumen of digestive tubules, as demonstrated by ultrastructural observations and electron probe X-ray microanalysis. TiO2 NPs of 60 nm were internalised within digestive cell lysosomes to a higher extent than TiO2 NPs of 180 nm, as confirmed by the quantification of black silver deposits after autometallography. The latter were localised mainly forming large aggregates in the lumen of the gut. Consequently, lysosomal membrane stability (LMS) was significantly reduced upon exposure to both TiO2 NPs although more markedly after exposure to TiO2-60 NPs. Exposure to bulk TiO2 and to DSLS also affected the stability of the lysosomal membrane. Thus, effects on the lysosomal membrane depended on the nanoparticle size and on the combined biological effects of TiO2 and DSLS.

  6. Engineering design of the Regolith X-ray Imaging Spectrometer (REXIS) instrument: an OSIRIS-REx student collaboration

    NASA Astrophysics Data System (ADS)

    Jones, Michael; Chodas, Mark; Smith, Matthew J.; Masterson, Rebecca A.

    2014-07-01

    OSIRIS-REx is a NASA New Frontiers mission scheduled for launch in 2016 that will travel to the asteroid Bennu and return a pristine sample of the asteroid to Earth. The REgolith X-ray Imaging Spectrometer (REXIS) is a student collaboration instrument on-board the OSIRIS-REx spacecraft. REXIS is a NASA risk Class D instrument, and its design and development is largely student led. The engineering team consists of MIT graduate and undergraduate students and staff at the MIT Space Systems Laboratory. The primary goal of REXIS is the education of science and engineering students through participation in the development of light hardware. In light, REXIS will contribute to the mission by providing an elemental abundance map of the asteroid and by characterizing Bennu among the known meteorite groups. REXIS is sensitive to X-rays between 0.5 and 7 keV, and uses coded aperture imaging to map the distribution of iron with 50 m spatial resolution. This paper describes the science goals, concept of operations, and overall engineering design of the REXIS instrument. Each subsystem of the instrument is addressed with a high-level description of the design. Critical design elements such as the Thermal Isolation Layer (TIL), radiation cover, coded-aperture mask, and Detector Assembly Mount (DAM) are discussed in further detail.

  7. Tumor-induced CD8+ T-cell dysfunction in lung cancer patients.

    PubMed

    Prado-Garcia, Heriberto; Romero-Garcia, Susana; Aguilar-Cazares, Dolores; Meneses-Flores, Manuel; Lopez-Gonzalez, Jose Sullivan

    2012-01-01

    Lung cancer is the leading cause of cancer deaths worldwide and one of the most common types of cancers. The limited success of chemotherapy and radiotherapy regimes have highlighted the need to develop new therapies like antitumor immunotherapy. CD8+ T-cells represent a major arm of the cell-mediated anti-tumor response and a promising target for developing T-cell-based immunotherapies against lung cancer. Lung tumors, however, have been considered to possess poor immunogenicity; even so, lung tumor-specific CD8+ T-cell clones can be established that possess cytotoxicity against autologous tumor cells. This paper will focus on the alterations induced in CD8+ T-cells by lung cancer. Although memory CD8+ T-cells infiltrate lung tumors, in both tumor-infiltrating lymphocytes (TILs) and malignant pleural effusions, these cells are dysfunctional and the effector subset is reduced. We propose that chronic presence of lung tumors induces dysfunctions in CD8+ T-cells and sensitizes them to activation-induced cell death, which may be associated with the poor clinical responses observed in immunotherapeutic trials. Getting a deeper knowledge of the evasion mechanisms lung cancer induce in CD8+ T-cells should lead to further understanding of lung cancer biology, overcome tumor evasion mechanisms, and design improved immunotherapeutic treatments for lung cancer.

  8. Evidence for a role of the PD-1:PD-L1 pathway in immune resistance of HPV-associated head and neck squamous cell carcinoma

    PubMed Central

    Lyford-Pike, Sofia; Peng, Shiwen; Young, Geoffrey D.; Taube, Janis M.; Westra, William H.; Akpeng, Belinda; Bruno, Tullia C.; Richmon, Jeremy D.; Wang, Hao; Bishop, Justin A.; Chen, Lieping; Drake, Charles G.; Topalian, Suzanne L.; Pardoll, Drew M.; Pai, Sara I.

    2013-01-01

    Human papillomavirus-associated head and neck squamous cell carcinomas (HPV-HNSCC) originate in the tonsils, the major lymphoid organ that orchestrates immunity to oral infections. Despite its location, the virus escapes immune elimination during malignant transformation and progression. Here, we provide evidence for the role of the PD-1:PD-L1 pathway in HPV-HNSCC immune resistance. We demonstrate membranous expression of PD-L1 in the tonsillar crypts, the site of initial HPV infection. In HPV-HNSCCs that are highly infiltrated with lymphocytes, PD-L1 expression on both tumor cells and CD68+ tumor associated macrophages (TAMs) is geographically localized to sites of lymphocyte fronts, while the majority of CD8+ tumor infiltrating lymphocytes (TILs) express high levels of PD-1, the inhibitory PD-L1 receptor. Significant levels of mRNA for interferon-γ (IFN-γ), a major cytokine inducer of PD-L1 expression, were found in HPV+ PD-L1(+) tumors. Our findings support the role of the PD-1:PD-L1 interaction in creating an “immune-privileged” site for initial viral infection and subsequent adaptive immune resistance once tumors are established and suggest a rationale for therapeutic blockade of this pathway in patients with HPV-HNSCC. PMID:23288508

  9. Propagation of gravity waves across the tropopause

    NASA Astrophysics Data System (ADS)

    Bense, Vera; Spichtinger, Peter

    2015-04-01

    The tropopause region is characterised by strong gradients in various atmospheric quantities that exhibit different properties in the troposphere compared to the stratosphere. The temperature lapse rate typically changes from negative to near-zero values resulting in a strong increase in stability. Accordingly, the buoyancy frequency often undergoes a jump at the tropopause. Analysis of radiosounding data also shows the existence of a strong inversion layer (tropopause inversion layer, TIL) characterised by a strong maximum in buoyancy frequency just above the tropopause, see e.g. Birner et al. (2002). Additionally, the magnitude of the vertical wind shear of the horizontal wind maximizes at the tropopause and the region also exhibits characteristical gradients of trace gases. Vertically propagating gravity waves can be excited in the troposphere by several mechanisms, e.g. by flow over topography (e.g. Durran, 1990), by jets and fronts (for a recent review: Plougonven and Zhang, 1990) or by convection (e.g. Clark et al., 1986). When these waves enter the tropopause region, their properties can be changed drastically by the changing stratification and strong wind shear. Within this work, the EULAG (Eulerian/semi-Lagrangian fluid solver, see e.g. Smolarkiewicz and Margolin, 1997) model is used to investigate the impact of the tropopause on vertically propagating gravity waves excited by flows over topography. The choice of topography (sine-shaped mountains, bell-shaped mountain) along with horizontal wind speed and tropospheric value of buoyancy frequency determine the spectrum of waves (horizontal and vertical wavelengths) that is excited in the tropsphere. In order to analyse how these spectra change for several topographies when a tropopause is present, we investigate different idealized cases in a two-dimensional domain. By varying the vertical profiles of buoyancy frequency (step-wise vs. continuos change, including TIL) and wind shear, the tropopause

  10. Subpicosecond Carrier Dynamics in Semiconductor Lasers and Lasers Based on Intersubband Transition.

    NASA Astrophysics Data System (ADS)

    Sung, Chun-Yung

    Recently, research for semiconductor lasers has proceeded in two directions. One direction is towards faster modulation speed for greater optical communication capacity. The second direction is towards long wavelength (10-100 mun) coherent radiation from the novel quantum well laser structures. In both cases, a fundamental understanding of carrier dynamics in the subpicosecond time scale is extremely important. This thesis explores the ultrafast carrier relaxation dynamics in several important optoelectronic semiconductor structures, with an emphasis on intersubband relaxation dynamics. Amplified femtosecond pulses are used for the generation of a femtosecond white light continuum, which has been used for the femtosecond differential transmission spectroscopy. We extensively studied the carrier relaxation for both low and high density excitation. The carrier capture, which is the intrinsic limit to the highest modulation bandwidth for SCH quantum well semiconductor lasers, has been time-resolved via femtosecond differential transmission spectroscopy. Strong capture resonances have been observed. Since the capture experiments reported to date do not directly measure the capture under conditions of laser operation, we have extended a three-pulse technique to remove carriers from the gain region of an inverted quantum well and monitor the gain recovery. This yields capture and relaxation times more appropriate for comparison with modulated laser operation. The gain recovery in a separate-confinement quantum-well structure limited by the carrier capture is directly time-resolved as well as spectral hole burning. Long carrier capture times are observed in the case of high density excitation due to Pauli-exclusion principle. The carrier tunneling dynamics in the new developed high speed tunneling injection laser (TIL) are also spectrally and temporally resolved. The measured ultrafast tunneling times confirm the TIL's potential for high speed applications. Ultrafast

  11. Halo-independent direct detection analyses without mass assumptions

    SciTech Connect

    Anderson, Adam J.; Fox, Patrick J.; Kahn, Yonatan; McCullough, Matthew

    2015-10-06

    Results from direct detection experiments are typically interpreted by employing an assumption about the dark matter velocity distribution, with results presented in the m{sub χ}−σ{sub n} plane. Recently methods which are independent of the DM halo velocity distribution have been developed which present results in the v{sub min}−g-tilde plane, but these in turn require an assumption on the dark matter mass. Here we present an extension of these halo-independent methods for dark matter direct detection which does not require a fiducial choice of the dark matter mass. With a change of variables from v{sub min} to nuclear recoil momentum (p{sub R}), the full halo-independent content of an experimental result for any dark matter mass can be condensed into a single plot as a function of a new halo integral variable, which we call h-til-tilde(p{sub R}). The entire family of conventional halo-independent g-tilde(v{sub min}) plots for all DM masses are directly found from the single h-tilde(p{sub R}) plot through a simple rescaling of axes. By considering results in h-tilde(p{sub R}) space, one can determine if two experiments are inconsistent for all masses and all physically possible halos, or for what range of dark matter masses the results are inconsistent for all halos, without the necessity of multiple g-tilde(v{sub min}) plots for different DM masses. We conduct a sample analysis comparing the CDMS II Si events to the null results from LUX, XENON10, and SuperCDMS using our method and discuss how the results can be strengthened by imposing the physically reasonable requirement of a finite halo escape velocity.

  12. [Immunotherapy of tumors of the central nervous system].

    PubMed

    Monod, L; Sawamura, Y; De Tribolet, N

    1992-01-01

    Malignant brain tumors are rapidly fatal and adjunction of chemo- and radio-therapy to surgical treatment has little changed their poor prognosis. The development of basic and clinical research in immunology of brain tumors has lead to new therapeutic strategies: 1) Humoral factors: Monoclonal antibodies (MAbs): alone or conjugated (to toxins, radionucleides or chemotherapeutic agents), MAbs can theoretically recognize antigens expressed by tumor cells and reach this target with high specificity. Large amounts of tumoricidal agents could be given to the tumor with low toxic effects to normal tissues. 2) Cellular factors: LAK cells: (lymphokine-activated-killer-cells) and activated TILs (tumor-infiltrating-lymphocytes) are autologous cytotoxic cells which can be produced by ex-vivo culture techniques and infused to the patient. These are very potent tumor-killer cells in vitro, however their in vivo effect is far less dramatic at the moment. 3) Cytokines: interferons, interleukins and other biological modifiers can act either directly on the tumor cells (cytotoxic effect) or indirectly through the modulation of the host-to-tumor response. 4) Combination of humoral and cellular factors: bispecific monoclonal antibodies are hybrid-molecules built from two different MAbs which can recognize two different targets, usually a tumor antigen on one hand and the T-cell-receptor (T 3) on the other hand. This combination of humoral and cellular effectors can theoretically lead to a preferential binding of effector cells to the tumor. All these new techniques are extensively studied in research laboratories and clinical trials without clear therapeutical benefit for the moment.

  13. Molecular cloning, tissue expression of gene Muc2 in blunt snout bream Megalobrama amblycephala and regulation after re-feeding

    NASA Astrophysics Data System (ADS)

    Xue, Chunyu; Xi, Bingwen; Ren, Mingchun; Dong, Jingjing; Xie, Jun; Xu, Pao

    2015-03-01

    Mucins are important components of mucus, which form a natural, physical, biochemical and semipermeable mucosal layer on the epidermis of fish gills, skin, and the gastrointestinal tract. As the first step towards characterizing the function of Muc2, we cloned a partial Megalobrama amblycephala Muc2 cDNA of 2 175 bp, and analyzed its tissue-specific expression pattern by quantitative real-time PCR (qPCR). The obtained sequence comprised 41 bp 5'-untranslated region (5'-UTR), 2 134 bp open reading frame encoding a protein of 711 amino acids. BLAST searching and phylogenetic analysis showed that the predicted protein contained several common secreted mucin-module domains (VWD-C8-TIL-VWD-C8) and had high homology with mucins from other vertebrates. Among four candidate reference genes ( β- Actin, RPI13α, RPII, 18S) for the qPCR, RPII was chosen as an appropriate reference gene because of its lowest variation in different tissues. M. amblycephala Muc2 was mainly expressed in the intestine, in the order (highest to lowest) middle-intestine > fore-intestine > hind-intestine. Muc2 was expressed relatively poorly in other organs (brain, liver, kidney, spleen, skin and gill). Furthermore, after 20-days of starvation, M. amblycephala Muc2 expressions after refeeding for 0 h, 3 h, 16 h, 3 d, and 10 d were significantly decreased in the three intestinal segments ( P<0.05) at 16 h, and were then upregulated to near the initial level at 10 d.

  14. Differential expression of immune-regulatory genes associated with PD-L1 display in melanoma: implications for PD-1 pathway blockade

    PubMed Central

    Taube, Janis M.; Young, Geoffrey D.; McMiller, Tracee L.; Chen, Shuming; Salas, January T.; Pritchard, Theresa S.; Xu, Haiying; Meeker, Alan K.; Fan, Jinshui; Cheadle, Chris; Berger, Alan E.; Pardoll, Drew M.; Topalian, Suzanne L.

    2015-01-01

    Purpose Blocking the immunosuppressive PD-1/PD-L1 pathway has anti-tumor activity in multiple cancer types, and PD-L1 expression on tumor cells and infiltrating myeloid cells correlates with the likelihood of response. We previously found that IFNG (interferon-gamma) was over-expressed by TILs in PD-L1+ vs. PD-L1(−) melanomas, creating adaptive immune resistance by promoting PD-L1 display. The current study was undertaken to identify additional factors in the PD-L1+ melanoma microenvironment coordinately contributing to immunosuppression. Experimental design Archived, formalin-fixed paraffin-embedded melanoma specimens were assessed for PD-L1 protein expression at the tumor cell surface with immunohistochemistry (IHC). Whole genome expression analysis, quantitative (q)RT-PCR, immunohistochemistry, and functional in vitro validation studies were employed to assess factors differentially expressed in PD-L1+ versus PD-L1(−) melanomas. Results Functional annotation clustering based on whole genome expression profiling revealed pathways up-regulated in PD-L1+ melanomas, involving immune cell activation, inflammation, and antigen processing and presentation. Analysis by qRT-PCR demonstrated over-expression of functionally related genes in PD-L1+ melanomas, involved in CD8+ T cell activation (CD8A, IFNG, PRF1, CCL5), antigen presentation (CD163, TLR3, CXCL1, LYZ), and immunosuppression [PDCD1 (PD-1), CD274(PD-L1), LAG3, IL10]. Functional studies demonstrated that some factors, including IL-10 and IL-32-gamma, induced PD-L1 expression on monocytes but not tumor cells. Conclusions These studies elucidate the complexity of immune checkpoint regulation in the tumor microenvironment, identifying multiple factors likely contributing to coordinated immunosuppression. These factors may provide tumor escape mechanisms from anti-PD-1/PD-L1 therapy, and should be considered for co-targeting in combinatorial immunomodulation treatment strategies. PMID:25944800

  15. Combined treatment with dabrafenib and trametinib with immune-stimulating antibodies for BRAF mutant melanoma

    PubMed Central

    Homet Moreno, Blanca; Mok, Stephen; Comin-Anduix, Begonya; Hu-Lieskovan, Siwen; Ribas, Antoni

    2016-01-01

    The combination of targeted therapy with BRAF and MEK inhibitors has become the standard of care in patients with BRAFV600E mutant melanoma, but responses are not durable. In addition, the impressive clinical benefits with anti-PD-1 and anti-PD-L1 antibodies (Ab) in patients with heavily pretreated metastatic melanoma and the synergistic effect of dabrafenib, trametinib and anti-PD-1 compared with single therapy alone groups support the idea that combining dabrafenib, trametinib and immunotherapy based on PD-1 blockade could be an interesting approach in the treatment of metastatic melanoma. With our mouse model of syngeneic BRAFV600E driven melanoma (SM1), we tested whether the addition of an immunostimulatory Ab targeting CD137 (4-1BB) and/or CD134 (OX40) would enhance the antitumor effect of dabrafenib, trametinib and anti-PD-1 or anti-PD-L1 therapy. In vitro studies showed that the combination group of dabrafenib, trametinib and anti-PD-1 increases CD8+ tumor infiltrating lymphocytes (TILs), as well as CD4+ T cells and tumor-associated macrophages (TAMs). An upregulation of PD-L1 was observed in the combination of dabrafenib, trametinib and anti-PD-1 therapy. Combination of dabrafenib, trametinib and anti-PD-1, with either anti-CD137 or anti-CD134, showed a superior antitumor effect, but the five-agent combination was not superior to the four-agent combinations. In conclusion, the combination of dabrafenib, trametinib, anti-PD1 or anti-PD-L1 therapy results in robust antitumor activity, which is further improved by adding the immune-stimulating Ab anti-CD137 or anti-CD134. Our findings support the testing of these combinations in patients with BRAFV600E mutant metastatic melanoma. PMID:27622011

  16. The classification and complex thermal history of the enstatite chondrites

    NASA Technical Reports Server (NTRS)

    Zhang, Yanhong; Benoit, Paul H.; Sears, Derek W. G.

    1995-01-01

    We have carried out instrumental neutron activation analysis of 11 enstatite chondrites and electron microprobe analyses of 17 enstatite chondrites, most of which were previously little described. We report here the third known EH5 chondrite (LEW 88180) and an unusual EL6 chondrite (LEW 87119), new data on four EL3 chondrites (ALH 85119, EET 90299, PCA 91020, and MAC 88136, which is paired with MAC 88180 and MAC 88184), the second EL5 chondrite (TIL 91714), and an unusual metal-rich and sulfide-poor EL3 chondrite (LEW 87223). The often discussed differences in mineral composition displayed by the EH and EL chondrites are not as marked after the inclusion of the new samples in the database, and the two classes apparently experienced a similar range of equilibrium temperatures. However, texturally the EL chondrites appear to have experienced much higher levels of metamorphic alteration than EH chondrites of similar equilibration temperatures. Most of the petrologic type criteria are not applicable to enstatite chondrites and, unlike the ordinary chondrites, texture and mineralogy reflect different aspects of the meteorite history. We therefore propose that the existing petrologic type scheme not be used for enstatite chondrites. We suggest that while 'textural type' reflects peak metamorphic temperatures, the 'mineralogical type' reflects equilibration during postmetamorphic (probably regolith) processes. Unlike the ordinary chondrites and EH chondrites, EL chondrites experienced an extensive low-temperature metamorphic episode. There are now a large number of enstatite meteorite breccias and impact melts, and apparently surface processes were important in determining the present nature of the enstatite chondrites.

  17. Patterns of bird functional diversity on land-bridge island fragments.

    PubMed

    Ding, Zhifeng; Feeley, Kenneth J; Wang, Yanping; Pakeman, Robin J; Ding, Ping

    2013-07-01

    The loss of species diversity due to habitat fragmentation has been extensively studied. In contrast, the impacts of habitat fragmentation on functional diversity remains relatively poorly understood. We conducted bird functional diversity studies on a set of 41 recently isolated land-bridge islands in the Thousand Island Lake, China. We analysed differences in bird species richness and a recently developed suite of complementary functional diversity indices (FRic, volume of functional space occupied; FEve, evenness of abundance distribution in the functional trait space; FDiv, divergence in the distribution of abundance in the trait volume) across different gradients (island area and isolation). We found no correlations between FRic and FEve or FEve and FDiv, but negative correlations between FRic and FDiv. As predicted, island area accounted for most of the variation in bird species richness, whereas isolation explained most of the variation in species evenness (decreasing species evenness with increasing isolation). Functional diversity appears to be more strongly influenced by habitat filtering as opposed to limiting similarity. More specifically, across all islands, both FRic and FEve were significantly lower than expected for randomly assembled communities, but FDiv showed no clear patterns. FRic increased with island area, FEve decreased with island area and FDiv showed no clear patterns. Our finding that FEve decreases with island area at TIL may indicate low functional stability on such islands, and as such large islands and habitat patches may deserve extra attention and/or protection. These results help to demonstrate the importance of considering the effects of fragmentation on functional diversity in habitat management and reserve design plans.

  18. Recombinant erythropoietin in humans has a prolonged effect on circulating erythropoietin isoform distribution.

    PubMed

    Aachmann-Andersen, Niels Jacob; Just Christensen, Søren; Lisbjerg, Kristian; Oturai, Peter; Meinild-Lundby, Anne-Kristine; Holstein-Rathlou, Niels-Henrik; Lundby, Carsten; Vidiendal Olsen, Niels

    2014-01-01

    The membrane-assisted isoform immunoassay (MAIIA) quantitates erythropoietin (EPO) isoforms as percentages of migrated isoforms (PMI). We evaluated the effect of recombinant human EPO (rhEPO) on the distribution of EPO isoforms in plasma in a randomized, placebo-controlled, double-blinded, cross-over study. 16 healthy subjects received either low-dose Epoetin beta (5000 IU on days 1, 3, 5, 7, 9, 11 and 13); high-dose Epoetin beta (30.000 IU on days 1, 2 and 3 and placebo on days 5, 7, 9, 11 and 13); or placebo on all days. PMI on days 4, 11 and 25 was determined by interaction of N-acetyl glucosamine with the glycosylation dependent desorption of EPO isoforms. At day 25, plasma-EPO in both rhEPO groups had returned to values not different from the placebo group. PMI with placebo, reflecting the endogenous EPO isoforms, averaged 82.5 (10.3) % (mean (SD)). High-dose Epoetin beta decreased PMI on days 4 and 11 to 31.0 (4.2)% (p<0.00001) and 45.2 (7.3)% (p<0.00001). Low-dose Epoetin beta decreased PMI on days 4 and 11 to 46.0 (12.8)% (p<0.00001) and 46.1 (10.4)% (p<0.00001). In both rhEPO groups, PMI on day 25 was still decreased (high-dose Epoetin beta: 72.9 (19.4)% (p=0.029); low-dose Epoetin beta: 73.1 (17.8)% (p=0.039)). In conclusion, Epoetin beta leaves a footprint in the plasma-EPO isoform pattern. MAIIA can detect changes in EPO isoform distribution up til at least three weeks after administration of Epoetin beta even though the total EPO concentration has returned to normal.

  19. New insights into the chemical structure of Y2Ti2O7-δ nanoparticles in oxide dispersion-strengthened steels designed for sodium fast reactors by electron energy-loss spectroscopy

    NASA Astrophysics Data System (ADS)

    Badjeck, V.; Walls, M. G.; Chaffron, L.; Malaplate, J.; March, K.

    2015-01-01

    In this paper we study by high resolution scanning transmission electron microscopy coupled with electron energy-loss spectroscopy (STEM-EELS) an oxide dispersion-strengthened (ODS) steel with the nominal composition Fe-14Cr-1W-0.3TiH2-0.3Y2O3 (wt.%) designed to withstand the extreme conditions met in Gen. IV nuclear reactors. After denoising via principal component analysis (PCA) the data are analyzed using independent component analysis (ICA) which is useful in the investigation of the physical properties and chemical structure of the material by separating the individual spectral responses. The Y-Ti-O nanoparticles are found to be homogeneously distributed in the ferritic matrix, sized from 1 to 20 nm and match a non-stoichiometric pyrochlore-Y2Ti2O7-δ structure for sizes greater than 5 nm. We show that they adopt a (Y-Ti-O)-Cr core-shell structure and that Cr also segregates at the matrix grain boundaries, which may slightly modify the corrosion properties of the steel. Using Ti-L2,3 and O-K fine structure (ELNES) the Ti oxidation state is shown to vary from the center of the nanoparticles to their periphery, from Ti4+ in distorted Oh symmetry to a valency often lower than 3+. The sensitivity of the Ti "white lines" ELNES to local symmetry distortions is also shown to be useful when investigating the strain induced in the nanoparticles by the surrounding matrix. The Cr-shell and the variation of the Ti valence state highlight a complex nanoparticle-matrix interface.

  20. Targeting metastatic cancer from the inside: a new generation of targeted gene delivery vectors enables personalized cancer vaccination in situ.

    PubMed

    Gordon, Erlinda M; Levy, John P; Reed, Rebecca A; Petchpud, W Nina; Liu, Liqiong; Wendler, Carlan B; Hall, Frederick L

    2008-10-01

    The advent of pathotropic (disease-seeking) targeting technologies, combined with advanced gene delivery vectors, provides a unique opportunity for the systemic delivery of immunomodulatory cytokine genes to remote sites of cancer metastasis. When injected intravenously, such pathotropic nanoparticles seek out and accumulate selectively at sites of tumor invasion and neo-angiogenesis, resulting in enhanced gene delivery, and thus cytokine production, within the tumor nodules. Used in conjunction with a primary tumoricidal agent (e.g., Rexin-G) that exposes tumor neoantigens, the tumor-targeted immunotherapy vector is intended to promote the recruitment and activation of host immune cells into the metastastic site(s), thereby initiating cancer immunization in situ. In this study, we examine the feasibility of cytokine gene delivery to cancerous lesions in vivo using intravenously administered pathotropically targeted nanoparticles bearing the gene encoding granulocyte/macrophage colony-stimulating factor (GM-CSF; i.e., Reximmune-C). In vitro, transduction of target cancer cells with Reximmune-C resulted in the quantitative production of bioactive and immunoreactive GM-CSF protein. In tumor-bearing nude mice, intravenous infusions of Reximmune-C-induced GM-CSF production by transduced cancer cells and paracrine secretion of the cytokine within the tumor nodules, which promoted the recruitment of host mononuclear cells, including CD40+ B cells and CD86+ dendritic cells, into the tumors. With the first proofs of principle established in preclinical studies, we generated an optimized vector configuration for use in advanced clinical trial designs, and extended the feasibility studies to the clinic. Targeted delivery and localized expression of the GM-CSF transgene was confirmed in a patient with metastatic cancer, as was the recruitment of significant tumor-infiltrating lymphocytes (TILs). Taken together, these studies provide the first demonstrations of cytokine gene