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At least 433 records · Page 24

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NASA NTRS1995

Gamma-interferon causes a selective induction of the lysosomal proteases, cathepsins B and L, in macrophages

Previous studies have indicated that acid-optimal cysteine proteinase(s) in the endosomal-lysosomal compartments, cathepsins, play a critical role in the proteolytic processing of endocytosed proteins to generate the antigenic peptides presented to the immune system on major histocompatibility complex (MHC) class II molecules. The presentation of these peptides and the expression of MHC class II molecules by macrophages and lymphocytes are stimulated by gamma-interferon (gamma-IFN). We found that treatment of human U-937 monocytes with gamma-IFN increased the activities and the content of the two major lysosomal cysteine proteinases, cathepsins B and L. Assays of protease activity, enzyme-linked immunosorbant assays (ELISA) and immunoblotting showed that this cytokine increased the amount of cathepsin B 5-fold and cathepsin L 3-fold in the lysosomal fraction. By contrast, the aspartic proteinase, cathepsin D, in this fraction was not significantly altered by gamma-IFN treatment. An induction of cathepsins B and L was also observed in mouse macrophages, but not in HeLa cells. These results suggest coordinate regulation in monocytes of the expression of cathepsins B and L and MHC class II molecules. Presumably, this induction of cysteine proteases contributes to the enhancement of antigen presentation by gamma-IFN.

Non-NASA Center↗

NASA NTRS1994

Negative regulators of cell proliferation

Cell proliferation is governed by the influence of both mitogens and inhibitors. Although cell contact has long been thought to play a fundamental role in cell cycling regulation, and negative regulators have long been suspected to exist, their isolation and purification has been complicated by a variety of technical difficulties. Nevertheless, over recent years an ever-expanding list of putative negative regulators have emerged. In many cases, their biological inhibitory activities are consistent with density-dependent growth inhibition. Most likely their interactions with mitogenic agents, at an intracellular level, are responsible for either mitotic arrest or continued cell cycling. A review of naturally occurring cell growth inhibitors is presented with an emphasis on those factors shown to be residents of the cell surface membrane. Particular attention is focused on a cell surface sialoglycopeptide, isolated from intact bovine cerebral cortex cells, which has been shown to inhibit the proliferation of an unusually wide range of target cells. The glycopeptide arrest cells obtained from diverse species, both fibroblasts and epithelial cells, and a broad variety of transformed cells. Signal transduction events and a limited spectrum of cells that are refractory to the sialoglycopeptide have provided insight into the molecular events mediated by this cell surface inhibitor.

Review↗

NASA NTRS1993

Overview of spaceflight immunology studies

The effects of spaceflight and analogues of spaceflight are discussed here and in nine accompanying articles. In this summary we present spaceflight studies with human subjects, animal subjects, and cell cultures and we review ground-based systems used to model the observed effects of spaceflight on the immune system. Human paradigms include bed rest, academic or psychological stress, physical stress, hypobaric or high altitude stress, and confinement. Animal models include antiorthostatic and orthostatic suspension, hypobarism, and confinement. The ten manuscripts in this collection were selected to provide a summary that should give the reader an overview of the various activities of spaceflight immunology researchers throughout the history of space travel. This manuscript identifies the major contributors to the study of spaceflight immunology, explains what types of studies have been conducted, and how they have changed over the years. Also presented is a discussion of the unusual limitations associated with spaceflight research and the efforts to develop appropriate ground-based surrogate model systems. Specific details, data, and mechanistic speculations will be held to a minimum, because they will be discussed in depth in the other articles in the collection.

NASA Discipline Number 18-10↗

NASA NTRS1993

The identification of a naturally occurring cell surface growth inhibitor related to a previously described bovine sialoglycopeptide

A 66-kDa sialoglycoprotein has been identified as the parental membrane molecule of an earlier described sialoglycopeptide (SGP), an 18-kDa molecule released by protease treatment of intact bovine cerebral cortex cells that was shown to be a potent inhibitor of cellular proliferation. The 66-kDa parental sialoglycoprotein (p-SGP) was purified approximately 2,400-fold, to apparent homogeneity, from bovine cerebral cortex cell membranes by its release during incubation with 3 M NaCl, preparative isoelectric focusing and lectin affinity chromatography. Although a membrane-associated molecule, the p-SGP appeared to be tightly bound to the cell membrane, since it was not released during incubations in the absence of 3 M NaCl. Incubation of the membrane preparations with 3 M urea proved to be too harsh, and the antigenicity required to follow the purification of the p-SGP was abolished. Analyses by SDS-PAGE, under reducing and nonreducing conditions, suggested that the p-SGP membrane component was a single polypeptide without subunit structure. The p-SGP was shown to be structurally related to the SGP fragment by immunoblots with IgG raised to the SGP inhibitor, and functionally related to the SGP by its ability to inhibit Swiss 3T3 proliferation at concentrations strikingly similar to that previous measured with the SGP fragment.

Non-NASA Center↗

NASA NTRS1992

Immunological characterization of pulmonary intravascular macrophages

Pulmonary intravascular macrophages (PIMs) are lung macrophages found apposed to the endothelium of pulmonary capillaries. In many species, they are responsible for the clearance of blood-borne particulates and pathogens; however, little else is known about their roles as immunologic effector cells. We compared PIMs with pulmonary alveolar macrophages (PAMs) to determine the relative immunological activities of these two cell populations. Our results suggested that both populations possess similar phagocytic and bactericidal activities. In assays measuring cytotoxicity, PIMs were more cytotoxic than PAMs against virally infected target cells; however, differences between these macrophage populations were not as marked when noninfected targets were used. LPS-stimulated PIMs produced more T-cell proliferative cytokines than PAMs, and both populations of nonstimulated macrophages produced similar amounts of the cytokines. In contrast, PAMs produced more TNF alpha and NO2- than PIMs when both populations were stimulated with LPS; however, nonstimulated PAMs and PIMs produced similar amounts of TNF alpha and NO2. These data suggest that bovine PIMs are immunologically active. Differences between the degrees of activity of PIMs and PAMs indicate that these macrophage populations may have different roles in lung surveillance.

NASA Discipline Cell Biology↗

NASA NTRS2004

Gene Expression Profile Analysis as a Prognostic Indicator of Normal Tissue Response to Simulated Space Radiations

This project was funded as a pilot project to determine the feasibility of using gene expression profiles to characterize the response of human cells to exposure to particulate radiations such as those encountered in the spaceflight environment. We proposed to use microarray technology to examine the gene expression patterns of a bank of well-characterized human fibroblast cell cultures. These fibroblast cultures were derived from breast or head and neck cancer patients who exhibited normal, minimal, or severe normal tissue reactions following low LET radiation exposure via radiotherapy. Furthermore, determination of SF2 values from fibroblasts cultured from these individuals were predictive of risk for severe late reactions. We hypothesized that by determining the expression of thousands of genes we could identify gene expression patterns that reflect how normal tissues respond to high Z and energy (HZE) particles, that is, that there are molecular signatures for HZE exposures. We also hypothesized that individuals who are intrinsically radiosensitive may elicit a unique response. Because this was funded as a pilot project we focused our initial studies on logistics and appropriate experimental design, and then to test our hypothesis that there is a unique molecular response to specific particles, in this case C and Fe, for primary human skin fibroblasts.

Story, Michael↗

NASA NTRS2004

Flow of Fluid and Particle Assemblages in Rotating Systems

NASA-designed bioreactors have been highly successful in growing three-dimensional tissue structures in a low shear environment both on earth and in space. The goal of the present study is to characterize the fluid flow environment within the HFB-S bioreactor and determine the spatial distribution of particles that mimic cellular tissue structures. The results will be used to obtain optimal operating conditions of rotation rates and media perfusehnfuse rates which are required for cell culture growth protocols. Two types of experiments have been performed so far. First, we have performed laser florescent dye visualization of the perfusion loop to determine the mixing times within the chamber. The second type of experiments involved particles which represent cellular tissue to determine the spatial distribution with the chamber. From these experiments we established that mixing times were largely dependant on the speed ratio and sign of the difference between the spinner and the dome. The shortest mixing times occurred when the spinner rotates faster than the dome and longest mixing times occurs with no relative motion between the dome and spinner. Also, we have determined the spatial and temporal distribution of particle assemblages within the chamber.

Kizito, John↗

NASA NTRS2005

Traction in smooth muscle cells varies with cell spreading

Changes in cell shape regulate cell growth, differentiation, and apoptosis. It has been suggested that the regulation of cell function by the cell shape is a result of the tension in the cytoskeleton and the distortion of the cell. Here we explore the association between cell-generated mechanical forces and the cell morphology. We hypothesized that the cell contractile force is associated with the degree of cell spreading, in particular with the cell length. We measured traction fields of single human airway smooth muscle cells plated on a polyacrylamide gel, in which fluorescent microbeads were embedded to serve as markers of gel deformation. The traction exerted by the cells at the cell-substrate interface was determined from the measured deformation of the gel. The traction was measured before and after treatment with the contractile agonist histamine, or the relaxing agonist isoproterenol. The relative increase in traction induced by histamine was negatively correlated with the baseline traction. On the contrary, the relative decrease in traction due to isoproterenol was independent of the baseline traction, but it was associated with cell shape: traction decreased more in elongated than in round cells. Maximum cell width, mean cell width, and projected area of the cell were the parameters most tightly coupled to both baseline and histamine-induced traction in this study. Wide and well-spread cells exerted larger traction than slim cells. These results suggest that cell contractility is controlled by cell spreading.

Non-NASA Center↗

NASA NTRS2007

The Lunar Environment: Determining the Health Effects of Exposure to Moon Dusts

The Earth s moon presents a hostile environment in which to live and work. There is no atmosphere to protect its surface from the ravages of solar wind and micrometeorite impacts. As a result, the moon s surface is covered with a thin layer of fine, charged, reactive dust capable of entering habitats and vehicle compartments, where it can result in crewmember health problems. During the Apollo missions, lunar dusts were introduced into the crew vehicle, resulting in direct exposure and occasional reports of respiratory, dermal and ocular irritation. In order to study the toxicological effects of lunar dust, NASA formed the Lunar Airborne Dust Toxicity Advisory Group (LADTAG). This interdisciplinary group is comprised of leading experts in space toxicology, lunar geology, space medicine and biomedical research. LADTAG has demonstrated that lunar soil contains several types of reactive dusts, including an extremely fine respirable component. These dusts have highly reactive surfaces in the lunar environment; the grains contain surface coatings which are generated by vapor phases formed by hypervelocity impact of micrometeorites. This unique class of dusts has surface properties that are unlike any Earth based analog. These distinctive properties are why lunar dusts are of great toxicological interest. Understanding how these reactive components behave "biochemically" in a moisture-rich pulmonary environment will aid in determining how toxic these particles are to humans. The data obtained from toxicological examination of lunar dusts will determine the human risk criteria for lunar dust exposure and produce a lunar health standard. LADTAG s analysis of lunar dusts and lunar dust simulants will include detailed lunar particle characterizations, determining the properties of particle activation, reactivation of lunar dust, the process of dust passivation and discerning the pathology of lunar dust exposure via inhalation, intratracheal instillation, cell culture exposure, dermal exposure and ocular exposure. The resulting health standard will be time-based and will vary by the duration and type of exposure. It may also be necessary to set multiple standards for different types of lunar dust, as well as for dust in its activated form vs. aged & passivated dust. This standard, set to protect the health of our robust astronaut crews, will not only impact NASA medical operations, but engineering designs as well. The data from our multidisciplinary research are vital in developing remediation devices and environmental monitors. Ultimately, the engineering and safety groups will design and develop countermeasures for space vehicles, suits, rovers and habitats that will be sustained within the limits of the health standard.

Khan-Mayberry, Noreen↗

NASA NTRS2007

Use of Atomic Oxygen for Increased Water Contact Angles of Various Polymers for Biomedical Applications

In the low Earth orbit (LEO) space environment, spacecraft surfaces can be altered during atomic oxygen exposure through oxidation and erosion. There can be terrestrial benefits of such interactions, such as the modification of hydrophobic or hydrophilic properties of polymers due to chemical modification and texturing. Such modification of the surface may be useful for biomedical applications. For example, atomic oxygen texturing may increase the hydrophilicity of polymers, such as chlorotrifluoroethylene (Aclar), thus allowing increased adhesion and spreading of cells on textured Petri dishes. The purpose of this study was to determine the effect of atomic oxygen exposure on the hydrophilicity of nine different polymers. To determine whether hydrophilicity remains static after atomic oxygen exposure or changes with exposure, the contact angles between the polymer and a water droplet placed on the polymer s surface were measured. The polymers were exposed to atomic oxygen in a radio frequency (RF) plasma asher. Atomic oxygen plasma treatment was found to significantly alter the hydrophilicity of non-fluorinated polymers. Significant decreases in the water contact angle occurred with atomic oxygen exposure. Fluorinated polymers were found to be less sensitive to changes in hydrophilicity for equivalent atomic oxygen exposures, and two of the fluorinated polymers became more hydrophobic. The majority of change in water contact angle of the non-fluorinated polymers was found to occur with very low fluence exposures, indicating potential cell culturing benefit with short treatment time.

Beger, Lauren↗

NASA NTRS2006

Technological Innovations from NASA

The challenge of human space exploration places demands on technology that push concepts and development to the leading edge. In biotechnology and biomedical equipment development, NASA science has been the seed for numerous innovations, many of which are in the commercial arena. The biotechnology effort has led to rational drug design, analytical equipment, and cell culture and tissue engineering strategies. Biomedical research and development has resulted in medical devices that enable diagnosis and treatment advances. NASA Biomedical developments are exemplified in the new laser light scattering analysis for cataracts, the axial flow left ventricular-assist device, non contact electrocardiography, and the guidance system for LASIK surgery. Many more developments are in progress. NASA will continue to advance technologies, incorporating new approaches from basic and applied research, nanotechnology, computational modeling, and database analyses.

Pellis, Neal R.↗

NASA NTRS2008

Hydrocyclone/Filter for Concentrating Biomarkers from Soil

The hydrocyclone-filtration extractor (HFE), now undergoing development, is a simple, robust apparatus for processing large amounts of soil to extract trace amounts of microorganisms, soluble organic compounds, and other biomarkers from soil and to concentrate the extracts in amounts sufficient to enable such traditional assays as cell culturing, deoxyribonucleic acid (DNA) analysis, and isotope analysis. Originally intended for incorporation into a suite of instruments for detecting signs of life on Mars, the HFE could also be used on Earth for similar purposes, including detecting trace amounts of biomarkers or chemical wastes in soils.

Ponce, Adrian↗

NASA NTRS2009

Optoelectronic pH Meter: Further Details

A collection of documents provides further detailed information about an optoelectronic instrument that measures the pH of an aqueous cell-culture medium to within 0.1 unit in the range from 6.5 to 7.5. The instrument at an earlier stage of development was reported in Optoelectronic Instrument Monitors pH in a Culture Medium (MSC-23107), NASA Tech Briefs, Vol. 28, No. 9 (September 2004), page 4a. To recapitulate: The instrument includes a quartz cuvette through which the medium flows as it is circulated through a bioreactor. The medium contains some phenol red, which is an organic pH-indicator dye. The cuvette sits between a light source and a photodetector. [The light source in the earlier version comprised red (625 nm) and green (558 nm) light-emitting diodes (LEDs); the light source in the present version comprises a single green- (560 nm)-or-red (623 nm) LED.] The red and green are repeatedly flashed in alternation. The responses of the photodiode to the green and red are processed electronically to obtain the ratio between the amounts of green and red light transmitted through the medium. The optical absorbance of the phenol red in the green light varies as a known function of pH. Hence, the pH of the medium can be calculated from the aforesaid ratio.

Jeevarajan, Antony S.↗

NASA NTRS2009

Molecular-specific urokinase antibodies

Antibodies have been developed against the different molecular forms of urokinase using synthetic peptides as immunogens. The peptides were synthesized specifically to represent those regions of the urokinase molecules which are exposed in the three-dimensional configuration of the molecule and are uniquely homologous to urokinase. Antibodies are directed against the lysine 158-isoleucine 159 peptide bond which is cleaved during activation from the single-chain (ScuPA) form to the bioactive double chain (54 KDa and 33 KDa) forms of urokinase and against the lysine 135 lysine 136 bond that is cleaved in the process of removing the alpha-chain from the 54 KDa form to produce the 33 KDa form of urokinase. These antibodies enable the direct measurement of the different molecular forms of urokinase from small samples of conditioned medium harvested from cell cultures.

Atassi, M. Zouhair↗

NASA NTRS2010

A Rat Body Phantom for Radiation Analysis

To reduce the uncertainties associated with estimating the biological effects of ionizing radiation in tissue, researchers rely on laboratory experiments in which mono-energetic, single specie beams are applied to cell cultures, insects, and small animals. To estimate the radiation effects on astronauts in deep space or low Earth orbit, who are exposed to mixed field broad spectrum radiation, these experimental results are extrapolated and combined with other data to produce radiation quality factors, radiation weighting factors, and other risk related quantities for humans. One way to reduce the uncertainty associated with such extrapolations is to utilize analysis tools that are applicable to both laboratory and space environments. The use of physical and computational body phantoms to predict radiation exposure and its effects is well established and a wide range of human and non-human phantoms are in use today. In this paper, a computational rat phantom is presented, as well as a description of the process through which that phantom has been coupled to existing radiation analysis tools. Sample results are presented for two space radiation environments.

Qualls, Garry D.↗

NASA NTRS2006

Micro-Imagers for Spaceborne Cell-Growth Experiments

A document discusses selected aspects of a continuing effort to develop five micro-imagers for both still and video monitoring of cell cultures to be grown aboard the International Space Station. The approach taken in this effort is to modify and augment pre-existing electronic micro-cameras. Each such camera includes an image-detector integrated-circuit chip, signal-conditioning and image-compression circuitry, and connections for receiving power from, and exchanging data with, external electronic equipment. Four white and four multicolor light-emitting diodes are to be added to each camera for illuminating the specimens to be monitored. The lens used in the original version of each camera is to be replaced with a shorter-focal-length, more-compact singlet lens to make it possible to fit the camera into the limited space allocated to it. Initially, the lenses in the five cameras are to have different focal lengths: the focal lengths are to be 1, 1.5, 2, 2.5, and 3 cm. Once one of the focal lengths is determined to be the most nearly optimum, the remaining four cameras are to be fitted with lenses of that focal length.

Behar, Alberto↗

NASA NTRS2000

Modeled Microgravity Inhibits Apoptosis in Peripheral Blood Lymphocytes

Microgravity interferes with numerous lymphocyte functions (expression of cell surface molecules, locomotion, polyclonal and antigen-specific activation, and the protein kinase C activity in signal transduction). The latter suggests that gravity may also affect programmed cell death (PCD) in lymphocyte populations. To test this hypothesis, we investigated spontaneous, activation- and radiation-induced PCD in peripheral blood mononuclear cells (PBMC) exposed to modeled microgravity using a rotating cell culture system. The results showed significant inhibition of radiation- and activation-induced apoptosis in modeled microgravity and provide insights into the potential mechanisms of this phenomenon.

Risin, Diana↗

NASA NTRS2000

Cellular Responses to Mechanical Stress Selected Contribution: A Three-Dimensional Model for Assessment of in Vitro Toxicity in Balaena Mysticetus Renal Tissue

This study established two- and three-dimensional renal proximal tubular cell cultures of the endangered species bowhead whale (Balaena mysticetus), developed SV40-transfected cultures, and cloned the 61-amino acid open reading frame for the metallothionein protein, the primary binding site for heavy metal contamination in mammals. Microgravity research, modulations in mechanical culture conditions (modeled microgravity), and shear stress have spawned innovative approaches to understanding the dynamics of cellular interactions, gene expression, and differentiation in several cellular systems. These investigations have led to the creation of ex vivo tissue models capable of serving as physiological research analogs for three-dimensional cellular interactions. These models are enabling studies in immune function, tissue modeling for basic research, and neoplasia. Three-dimensional cellular models emulate aspects of in vivo cellular architecture and physiology and may facilitate environmental toxicological studies aimed at elucidating biological functions and responses at the cellular level. Marine mammals occupy a significant ecological niche (72% of the Earth's surface is water) in terms of the potential for information on bioaccumulation and transport of terrestrial and marine environmental toxins in high-order vertebrates. Few ex vivo models of marine mammal physiology exist in vitro to accomplish the aforementioned studies. Techniques developed in this investigation, based on previous tissue modeling successes, may serve to facilitate similar research in other marine mammals.

Goodwin, T. J.↗