A radar data processing and enhancement system

This report describes the space position data processing system of the NASA Western Aeronautical Test Range. The system is installed at the Dryden Flight Research Facility of NASA Ames Research Center. This operational radar data system (RADATS) provides simultaneous data processing for multiple data inputs and tracking and antenna pointing outputs while performing real-time monitoring, control, and data enhancement functions. Experience in support of the space shuttle and aeronautical flight research missions is described, as well as the automated calibration and configuration functions of the system

Phenotypic hypersusceptibility to multiple protease inhibitors and low replicative capacity in patients who are chronically infected with human immunodeficiency virus type 1

Increased susceptibility to the protease inhibitors saquinavir and amprenavir has been observed in human immunodeficiency virus type 1 (HIV-1) with specific mutations in protease (V82T and N88S). Increased susceptibility to ritonavir has also been described in some viruses from antiretroviral agent-naïve patients with primary HIV-1 infection in association with combinations of amino acid changes at polymorphic sites in the protease. Many of the viruses displaying increased susceptibility to protease inhibitors also had low replication capacity. In this retrospective study, we analyze the drug susceptibility phenotype and the replication capacity of virus isolates obtained at the peaks of viremia during five consecutive structured treatment interruptions in 12 chronically HIV-1-infected patients. Ten out of 12 patients had at least one sample with protease inhibitor hypersusceptibility (change ≤0.4-fold) to one or more protease inhibitor. Hypersusceptibility to different protease inhibitors was observed at variable frequency, ranging from 38% to amprenavir to 11% to nelfinavir. Pairwise comparisons between susceptibilities for the protease inhibitors showed a consistent correlation among all pairs. There was also a significant relationship between susceptibility to protease inhibitors and replication capacity in all patients. Replication capacity remained stable over the course of repetitive cycles of structured treatment interruptions. We could find no association between in vitro replication capacity and in vivo plasma viral load doubling time and CD4(+) and CD8(+) T-cell counts at each treatment interruption. Several mutations were associated with hypersusceptibility to each protease inhibitor in a univariate analysis. This study extends the association between hypersusceptibility to protease inhibitors and low replication capacity to virus isolated from chronically infected patients and highlights the complexity of determining the genetic basis of this phenomenon. The potential clinical relevance of protease inhibitor hypersusceptibility and low replication capacity to virologic response to protease inhibitor-based therapies deserves to be investigated further

PEMBELAJARAN TEKNIK JUMPUTAN DI KELAS X IPA 1 SMAN 1 SEWON

Tujuan penelitian ini adalah: 1) mengetahui persiapan pembelajaran teknik jumputan; 2) mengetahui pelaksanaan pembelajaran teknik jumputan; 3) mengetahui hasil pembelajaran jumputan. Penelitian ini merupakan penelitian deskriptif kualitatif. Data dalam penelitian ini berupa kata-kata dan tindakan yang diperoleh dengan observasi, wawancara dan dokumentasi. Instrumen utama dalam penelitian ini yaitu peneliti sendiri dibantu dengan pedoman observasi, pedoman wawancara, pedoman dokumentasi dan menggunakan alat bantu kamera serta alat tulis. Teknik analisis data menggunakan deskriptif kualitatif yang dilakukan dengan tahapan reduksi data, penyajian data dan penarikan kesimpulan. Untuk mengecek keabsahan data menggunakan teknik ketekunan pengamatan dan perpanjangan pengamatan. Hasil penelitian adalah sebagai berikut. 1) Proses pembelajaran teknik jumputan di SMAN 1 Sewon dimulai dengan melakukan beberapa persiapan diantaranya membuat silabus, RPP, dan bahan ajar. 2) Proses pembelajaran terdiri dari kegiatan pendahuluan, inti dan penutup. Kegiatan pendahuluan merupakan kegiatan yang dilakukan sebelum memasuki materi pembelajaran. Kegiatan inti adalah kegiatan pokok pembelajaran yang berisi materi yang diajarkan. Dalam kegiatan penutup guru memberikan kesimpulan yang diperoleh dari kegiatan pembelajaran yang telah dilakukan serta menanamkan nilai-nilai seperti sikap tanggung jawab, jujur, kreatif dan percaya diri. 3) Hasil penilaian karya siswa menunjukkan bahwa pembelajaran teknik jumputan di kelas X IPA 1 dikatakan berhasil. Hal tersebut dapat dilihat dari nilai 30 siswa yang berada di atas KSM sekolah sebesar 76. 4) Hasil pembelajaran teknik jumputan di kelas X IPA 1 adalah pola, motif jumputan, dan produk serta nilai hasil belajar teknik jumputan

Development and Evaluation of Mouse Monoclonal Antibodies Against Human C1q

The complement protein C1q plays an important role in breast cancer susceptibility, development, and progression. Mice genetically deficient in C1qA exhibit an eighty-five percent decrease in mammary tumour incidence when administered the chemical carcinogen DMBA. Similarly, in the aggressive MMTV-PyMT model of mouse mammary cancer, C1qA null mutant mice exhibit two thirds of the tumour burden seen in wild type mice and a third of the late-stage carcinoma at eighteen weeks of age. There may also be a role for C1q in human breast cancer as women genetically deficient in C1q have a decreased incidence of breast cancer. C1q plays a key role in macrophage-mediated efferocytic uptake of dying mammary epithelial cells, thereby preventing apoptotic cells from becoming necrotic and releasing pro-inflammatory cytoplasmic components. Persistence of necrotic cells alters the immune response during cancer initiation, as evidenced by an increase in cytotoxic T cells mobilised to the mammary gland of C1q null mutant mice in response to carcinogen DMBA. The over-arching goal of this research project was to generate and characterise an inhibitor of C1q-mediated efferocytosis. A monoclonal antibody was chosen as the C1q inhibitor, to further explore the role of C1q in breast cancer and as a potential first step in development of a new breast cancer therapeutic. Human C1q was inoculated into C1qA null mutant mice to generate an antibody-mediated immune response to C1q. Three fusions of immune splenocytes from these mice yielded a total of 2,776 cultures of which 2,017 contained viable hybridomas. Antibody binding by enzyme-linked immunosorbent assay (ELISA) included both linear epitopes (contiguous amino acids) and conformational epitopes (binding to a three-dimensional structure), making this an ideal screening strategy. Screening of these cultures by ELISA identified eight hybridomas that bound C1q. Of these, four were successfully expanded and cultures established from single cells. Thus, four candidate monoclonal antibodies were generated: BHI1-1G4, BHI1-4D3, BHI3-3F6, and BHI3-8B9. Characterisation of these antibodies was performed to determine the specificity of their binding conditions. Binding of the monoclonal antibodies in assays that involve denaturation of proteins and presentation of linear epitopes was not observed. These assays included western blotting, immunohistochemistry on normal breast sections, and immunocytochemistry on a fixed macrophage cell line. An assay with potential to display conformational epitopes, immunocytochemistry on unfixed macrophages, also did not demonstrate monoclonal binding. The antibodies were also tested for binding to C1q by immunoprecipitation. This assay can detect conformational epitopes, and all four monoclonal antibodies were demonstrated to bind soluble C1q by this method. Combined, these studies suggested that all four candidate monocloncal antibodies bind only to intact native C1q and do not bind to denatured antigen. Identification of a monoclonal antibody that inhibits C1q-mediated efferocytosis required development of a bioassay that quantifies the functional activity of candidate antibodies. An in vitro efferocytosis assay was developed involving fluorescent green-labelled macrophages co-cultured with fluorescent red-labelled MDA-MB-231 breast cancer cells induced to undergo apoptosis by cross-linking the TRAIL receptor 2. Co-localisation of red and green fluorescence as an indicator of efferocytosis was investigated in bone marrow-derived macrophages from C1qA replete and null mice. Reduced efferocytosis was observed in macrophages where C1q was absent and was quantified using ImageJ software involving digital masking of images and Boolean calculation of overlap. Hybridoma supernatants from candidate antibodies BHI1-1G4 and BHI1-4D3 were tested in the assay using the mouse macrophage cell line RAW 264.7 labelled green and dying breast cancer cells labelled red. Commercially available anti-C1q antibody 9A7 and hybridoma supernatant from non-C1q binding cell line BHI3-2C12 were also assessed. The antibodies exhibited variable capacity to affect C1q-mediated phagocytosis however whether a specific candidate monoclonal antibody could effectively inhibit C1q was inconclusive due to a high degree of variability in the timing of apoptotic cell uptake. This research led to the generation of four candidate monoclonal antibodies with potential for further pre-clinical research. Future work should concentrate on purification of the antibodies in order to improve investigation of their inhibitory capacity in C1q-mediated efferocytosis bioassays. Studies in mouse mammary cancer models would also provide valuable data on the potential of these candidate antibodies for downstream clinical applications in breast cancer patients.Thesis (Ph.D.) -- University of Adelaide, Adelaide Medical School, 202

Susceptibility Provision Enhances Effective De-escalation (SPEED): utilizing rapid phenotypic susceptibility testing in Gram-negative bloodstream infections and its potential clinical impact

Abstract Objectives We evaluated the performance and time to result for pathogen identification (ID) and antimicrobial susceptibility testing (AST) of the Accelerate Pheno™ system (AXDX) compared with standard of care (SOC) methods. We also assessed the hypothetical improvement in antibiotic utilization if AXDX had been implemented. Methods Clinical samples from patients with monomicrobial Gram-negative bacteraemia were tested and compared between AXDX and the SOC methods of the VERIGENE® and Bruker MALDI Biotyper® systems for ID and the VITEK® 2 system for AST. Additionally, charts were reviewed to calculate theoretical times to antibiotic de-escalation, escalation and active and optimal therapy Results ID mean time was 21 h for MALDI-TOF MS, 4.4 h for VERIGENE® and 3.7 h for AXDX. AST mean time was 35 h for VITEK® 2 and 9.0 h for AXDX. For ID, positive percentage agreement was 95.9% and negative percentage agreement was 99.9%. For AST, essential agreement was 94.5% and categorical agreement was 93.5%. If AXDX results had been available to inform patient care, 25% of patients could have been put on active therapy sooner, while 78% of patients who had therapy optimized during hospitalization could have had therapy optimized sooner. Additionally, AXDX could have reduced time to de-escalation (16 versus 31 h) and escalation (19 versus 31 h) compared with SOC. Conclusions By providing fast and reliable ID and AST results, AXDX has the potential to improve antimicrobial utilization and enhance antimicrobial stewardship

Identification of a Cell Surface Protein with a Role in Stimulating Human Keratinocyte Proliferation, Expressed During Development and Carcinogenesis

In an attempt to define cell surface molecules with an important role in the development of squamous cell carcinomas (SCCs), we generated monoclonal antibodies (MoAbs) to a human keratinocyte cell line (FEP18-11-T1) capable of giving rise to SCCs in nude mice. MoAb 10G7 was selected for further study because it bound to a cell surface component preferentially expressed by this cell line as compared with normal human foreskin keratinocytes. This MoAb recognizes a cell surface protein (10G7 antigen) that is not detectable on normal keratinocytes in the foreskin in vivo, but whose expression is induced when the keratinocytes are dissociated from this tissue and placed in culture. Interestingly, the 10G7 antigen is downregulated upon keratinocyte differentiation in vitro. Consistent with its expression in hyper-proliferative epithella in vitro, 10G7 antigen exhibited a classic oncofetal pattern of expression in vivo. Thus, although no reactivity was obtained with MoAb 10G7 in the epithelia of normal foreskin or cervical tissue, strong reactivity was detected in epithelia from genital lesions ranging from benign warts to invasive SCCs. Epidermis from developing fetal tissue also exhibited strong reactivity with MoAb 10G7. We have been able to demonstrate that this MoAb is capable of stimulating FEP18–11-T1 keratinocyte proliferation in vitro in a concentration-dependent manner in the absence of growth factors, suggesting that the 10G7 antigen may play an important role in regulating cellular proliferation during development and in carcinogenesis in epithelial tissues