Nutrient requirements and other factors involved in the culture of human kidney cells on microcarrier beads

The culture of human kidney cells on microcarrier beads in the Bioprocessing Laboratory at the Johnson Space Center is described. These were the first series of studies performed before and during 1983 to determine optimum conditions, including medium type, bead type and density. The composition of several medium types and the molecular weights of some common culture medium supplements and cellular proteins are included. The microgravity cell-to-bead attachment experiment performed on Space Transportation System Flight 8 is described

Margaret Chase Smith’s 1950 Declaration of Conscience Speech

In 1948 Margaret Chase Smith of Maine became the first woman elected to the Senate entirely on her own merit. She went on to enjoy a long and distinguished career in Congress. The highlight of this career was Smith’s 1950 speech against foe McCarthy, known as her Declaration of Conscience. In the following article, Dennis Morrison analyzes the speech and traces its origins to Smith’s early life in Maine

Appropriate Selection Processes Are Available For Choosing Hospitality texts

First Student: I know what you mean; I fall asleep every time I try to read this stuff. (Meanwhile, in the faculty lounge) First Professor: I do not understand what\u27s wrong with my students; they expect me to teach textbook information from the podium. Second Professor: I\u27ve noticed the same thing. They do not want to be responsible for their readings

Biochemical assays on plasminogen activators and hormones from kidney sources

Investigations were established for the purpose of analyzing the conditioned media from human embryonic kidney cell subpopulations separated in space by electrophoresis. This data is based on the experiments performed on STS-8 on the continuous flow electrophoresis system. The primary biological activity that was analyzed was plasminogen activator activity, but some assays for erythropoeitin and human granulocyte colony stimulating activity were also performed. It is concluded that a battery of assays are required to completely define the plasminogen activator profile of a conditioned media from cell culture. Each type of assay measures different parts of the mixture and are influenced by different parameters. The functional role of each assay is given along with an indication of which combination of assays are required to answer specific questions. With this type of information it is possible by combinations of assays with mathematical analysis to pinpoint a specific component of the system

Geologic factors in the evaluation of water pollution potential at mountain dwelling sites

December, 1973.Bibliography: pages [133]-135.In order to establish the relationship between the geologic setting and the occurrence of water pollution in mountain home developments containing individual sewage disposal systems, three areas in Colorado's Front Range were studied. Two of the areas were known to have biological contamination as confirmed by microbiological tests. Also, each area had adequate rock exposures to allow for detailed geologic study, the absence of thick soil profiles above bedrock and current development for mountain home location. In addition, each of the selected study areas differed in geologic setting, age of development and home density in the development. The extent of water pollution was established by a program of well and surface water testing for total coliform. Fecal coliform and inorganic contaminants were tested in selected wells. Detailed geologic maps were made of each area to locate features such as dikes or shear zones which might act as either barriers or conduits to ground water movement. Slope maps were prepared for the area by computer plots of digitized data of elevations taken from U.S. Geological Survey topographic maps. Determination of the water table profile and extent of alluvial fill in valley bottoms was accomplished using driller's well logs for each well in the area. In addition, soils were tested to determine their effective grain size, a joint and foliation study was conducted to determine the direction of pollutant travel should effluent enter these openings, driller's well logs were used to establish depth of soils and depth of weathering in bedrock and data from county health records were used to establish soil percolation rates. Data were compiled in the form of overlays on base maps of the areas involved. A topographic map with the geologic overlay was used in conjunction with various combinations of the field derivative overlays to indicate the pollution potential for specific areas. The overlays used in this procedure were compiled from. the following parameters: 1) slope, 2) depth of soil, 3) depth of intensely weathered bedrock, 4) local water table profile, and 5) soil percolation rates. These overlays indicated that the Glen Haven area is unsuitable for soil absorption sewage systems because of steep slopes, soil depth and depth of the water table. Most of the Tall Timbers area was indicated as unsuitable for soil absorption systems because of slope, soil depth and local geology. The Crescent Park area was categorized as safe (in part) for soil absorption systems, however, local areas within the subdivision were categorized as hazardous. From the results it was suggested that a procedure such as the one used in this study could be used for each subdivision proposed in the mountainous regions of Colorado. Thus more effective use of mountainous areas might be passable while maintaining a low probability of ground water contamination. Areas within each proposed subdivision would be classified as safe, hazardous, or unsuitable for soil absorption sewage systems. Unsuitable areas could be used as parks or greenbelts, hazardous areas would have low population densities and safe areas would be allowed to have higher population densities as long as other factors were favorable. In addition, procedures such as the one used in this investigation could be used to indicate mountain areas which should require a municipal sewage disposal system before development to ensure that the ground water system was not polluted.Supported by the Colorado Water Quality Control Commission and the United States Department of Interior, Office of Water Resources Research, no.87-194-2. (s) 14-01-001-1882

Kidney cell electrophoresis in space flight: Rationale, methods, results and flow cytometry applications

Cultures of human embryonic kidney cells consistently contain an electrophoretically separable subpopulation of cells that produce high levels of urokinase and have an electrophoretic mobility about 85 percent as high as that of the most mobile human embryonic kidney cells. This subpopulation is rich in large epithelioid cells that have relatively little internal structure. When resolution and throughput are adequate, free fluid electrophoresis can be used to isolate a broad band of low mobility cells which also produces high levels of plasminogen activators (PAs). In the course of performing this, it was discovered that all electrophoretic subpopulations of cultured human embryonic kidney cells produce some PAs and that separate subpopulations produce high quantities of different types of PA's. This information and the development of sensitive assays for this project have provided new insights into cell secretion mechanisms related to fibrinolysis. These advances would probably not have been made without the NASA program to explore fundamental questions of free fluid electrophoresis in space

Activation of Penile Proadipogenic Peroxisome Proliferator-Activated Receptor γ with an Estrogen: Interaction with Estrogen Receptor Alpha during Postnatal Development

Exposure to the estrogen receptor alpha (ERα) ligand diethylstilbesterol (DES) between neonatal days 2 to 12 induces penile adipogenesis and adult infertility in rats. The objective of this study was to investigate the in vivo interaction between DES-activated ERα and the proadipogenic transcription factor peroxisome proliferator-activated receptor gamma (PPARγ). Transcripts for PPARs α, β, and γ and γ1a splice variant were detected in Sprague-Dawley normal rat penis with PPARγ predominating. In addition, PPARγ1b and PPARγ2 were newly induced by DES. The PPARγ transcripts were significantly upregulated with DES and reduced by antiestrogen ICI 182, 780. At the cellular level, PPARγ protein was detected in urethral transitional epithelium and stromal, endothelial, neuronal, and smooth muscular cells. Treatment with DES activated ERα and induced adipocyte differentiation in corpus cavernosum penis. Those adipocytes exhibited strong nuclear PPARγ expression. These results suggest a biological overlap between PPARγ and ERα and highlight a mechanism for endocrine disruption

MTBVAC vaccination protects rhesus macaques against aerosol challenge with M. tuberculosis and induces immune signatures analogous to those observed in clinical studies

A single intradermal vaccination with MTBVAC given to adult rhesus macaques was well tolerated and conferred a significant improvement in outcome following aerosol exposure to M. tuberculosis compared to that provided by a single BCG vaccination. Vaccination with MTBVAC resulted in a significant reduction in M. tuberculosis infection-induced disease pathology measured using in vivo medical imaging, in gross pathology lesion counts and pathology scores recorded at necropsy, the frequency and severity of pulmonary granulomas and the frequency of recovery of viable M. tuberculosis from extrapulmonary tissues following challenge. The immune profiles induced following immunisation with MTBVAC reflect those identified in human clinical trials of MTBVAC. Evaluation of MTBVAC- and TB peptide-pool-specific T-cell cytokine production revealed a predominantly Th1 response from poly- (IFN-¿+TNF-a+IL2+) and multi-(IFN-¿+TNF-a+) functional CD4 T cells, while only low levels of Th22, Th17 and cytokine-producing CD8 T-cell populations were detected together with low-level, but significant, increases in CFP10-specific IFN-¿ secreting cells. In this report, we describe concordance between immune profiles measured in clinical trials and a macaque pre-clinical study demonstrating significantly improved outcome after M. tuberculosis challenge as evidence to support the continued development of MTBVAC as an effective prophylactic vaccine for TB vaccination campaigns

IV BCG Vaccination and Aerosol BCG Revaccination Induce Mycobacteria-Responsive γδ T Cells Associated with Protective Efficacy against M. tb Challenge.

Intravenously (IV) delivered BCG provides superior tuberculosis (TB) protection compared with the intradermal (ID) route in non-human primates (NHPs). We examined how γδ T cell responses changed in vivo after IV BCG vaccination of NHPs, and whether these correlated with protection against aerosol M. tuberculosis challenge. In the circulation, Vδ2 T cell populations expanded after IV BCG vaccination, from a median of 1.5% (range: 0.8-2.3) of the CD3+ population at baseline, to 5.3% (range: 1.4-29.5) 4 weeks after M. tb, and were associated with TB protection. This protection was related to effector and central memory profiles; homing markers; and production of IFN-γ, TNF-α and granulysin. In comparison, Vδ2 cells did not expand after ID BCG, but underwent phenotypic and functional changes. When Vδ2 responses in bronchoalveolar lavage (BAL) samples were compared between routes, IV BCG vaccination resulted in highly functional mucosal Vδ2 cells, whereas ID BCG did not. We sought to explore whether an aerosol BCG boost following ID BCG vaccination could induce a γδ profile comparable to that induced with IV BCG. We found evidence that the aerosol BCG boost induced significant changes in the Vδ2 phenotype and function in cells isolated from the BAL. These results indicate that Vδ2 population frequency, activation and function are characteristic features of responses induced with IV BCG, and the translation of responses from the circulation to the site of infection could be a limiting factor in the response induced following ID BCG. An aerosol boost was able to localise activated Vδ2 populations at the mucosal surfaces of the lung. This vaccine strategy warrants further investigation to boost the waning human ID BCG response