Bobby L. Wilson

Analysis of stress responsive genes induced by single-walled carbon nanotubes in BJ Foreskin cells.

It is known that the mechanical properties of clay-reinforced nanocomposites are significantly affected by the dispersion of clay particles in the matrix. In this study, the effect of surface-treatment of Montmorillonite (MMT) on the fracture behavior of MMT/epoxy nanocomposite was investigated. For this purpose, fracture tests were performed using samples with three different clay concentration level. After fracture tests, SEM analysis was made on the fracture surfaces to examine the fracture mechanism. It was found that the MMT treatment using 3-aminopropyltriethoxysilane enhanced the fracture toughness increased of the MMT/epoxy nanocomposite. This is due to the improved intercalation effect and interfacial strength between MMT and epoxy matrix.

ACTIVATION OF NUCLEAR TRANSCRIPTION FACTOR–κB IN MOUSE BRAIN INDUCED BY A SIMULATED MICROGRAVITY ENVIRONMENT

SummaryMicrogravity induces inflammatory responses and modulates immune functions that may increase oxidative stress. Exposure to a microgravity environment induces adverse neurological effects; however, there is little research exploring the etiology of these effects resulting from exposure to such an environment. It is also known that spaceflight is associated with increase in oxidative stress; however, this phenomenon has not been reproduced in land-based simulated microgravity models. In this study, an attempt has been made to show the induction of reactive oxygen species (ROS) in mice brain, using ground-based microgravity simulator. Increased ROS was observed in brain stem and frontal cortex with concomitant decrease in glutathione, on exposing mice to simulated microgravity for 7 d. Oxidative stress-induced activation of nuclear factor-kappaB was observed in all the regions of the brain. Moreover, mitogen-activated protein kinase kinase was phosphorylated equally in all regions of the brain exposed to simulated microgravity. These results suggest that exposure of brain to simulated microgravity can induce expression of certain transcription factors, and these have been earlier argued to be oxidative stress dependent.

Pulmonary biocompatibility assessment of inhaled single-wall and multiwall carbon nanotubes in BALB/c mice.

With the widespread application of carbon nanotubes (CNTs) in diverse commercial processes, scientists are now concerned about the potential health risk of occupational exposures. In this study, CNT-induced pulmonary toxicity was investigated by exposing BALB/c mice to aerosolized single-wall (SW) CNT and multiwall (MW) CNT (5 μg/g of mice) for 7 consecutive days in a nose-only exposure system. Microscopic studies showed that inhaled CNTs were homogeneously distributed in the mouse lung. The total number of bronchoalveolar lavage polymorphonuclear leukocytes recovered from the mice exposed to SWCNT and MWCNT (1.2 × 106 ± 0.52 and 9.87 × 105 ± 1.45; respectively) was significantly greater than control mice (5.46 × 105 ± 0.78). Rapid development of pulmonary fibrosis in mice that inhaled CNT was also confirmed by significant increases in the collagen level. The lactate dehydrogenase levels were increased nearly 2- and 2.4-fold in mice that inhaled SWCNT and MWCNT, respectively, as compared with control mice. In addition, exposure of CNTs to mice showed a significant (p < 0.05) reduction of antioxidants (glutathione, superoxide dismutase, and catalase) and induction of oxidants (myloperoxidase, oxidative stress, and lipid peroxidation) compared with control. Apoptosis-related proteins such as caspase-3 and -8 activities were also significantly increased in mice that inhaled CNT than in control mice. Together, this study shows that inhaled CNTs induce inflammation, fibrosis, alteration of oxidant and antioxidant levels, and induction of apoptosis-related proteins in the lung tissues to trigger cell death.

Proteomic analysis of mouse hypothalamus under simulated microgravity.

Exposure to altered microgravity during space travel induces changes in the brain and these are reflected in many of the physical behavior seen in the astronauts. The vulnerability of the brain to microgravity stress has been reviewed and reported. Identifying microgravity-induced changes in the brain proteome may aid in understanding the impact of the microgravity environment on brain function. In our previous study we have reported changes in specific proteins under simulated microgravity in the hippocampus using proteomics approach. In the present study the profiling of the hypothalamus region in the brain was studied as a step towards exploring the effect of microgravity in this region of the brain. Hypothalamus is the critical region in the brain that strictly controls the pituitary gland that in turn is responsible for the secretion of important hormones. Here we report a 2-dimensional gel electrophoretic analysis of the mouse hypothalamus in response to simulated microgravity. Lowered glutathione and differences in abundance expression of seven proteins were detected in the hypothalamus of mice exposed to microgravity. These changes included decreased superoxide dismutase-2 (SOD-2) and increased malate dehydrogenase and peroxiredoxin-6, reflecting reduction of the antioxidant system in the hypothalamus. Taken together the results reported here indicate that oxidative imbalance occurred in the hypothalamus in response to simulated microgravity.

Altered cytokine expression in tissues of mice subjected to simulated microgravity.

Space flight is known to induce microgravity-associated immune dysfunction in humans, non-human primates and rodents. To understand the mechanism underlying these defects, several studies in rodents have been conducted in a ground-based antiorthostatic suspension (AOS) model that would mimic the effects of microgravity. In all these in vivo studies that showed the effects on cytokine profiles actually investigated the ex vivo production from culturing the cells isolated from whole organism that was exposed to space flight and/or microgravity. So, the purpose of the study was to examine the in vivo expression of cytokines in mice in immunologically important tissue environments of mice that were subjected to AOS. Cytokines such as Interleukin-1β (IL-1β), IL-2, IL-3, IL-6, Interferon-γ (IFN-γ) and Tumor Necrosis Factor-α (TNF-α) were measured by Enzyme Linked Immunosorbent Assay (ELISA) in the homogenates of spleen tissue, lymph nodes and also in serum of AOS mice and compared with that of control mice. AOS induced no change in the IL-3 levels, but IL-1β was increased significantly whereas IL-2 levels decreased in spleen, lymph nodes and serum. IL-6 levels did not differ in spleen but were significantly increased in lymph nodes and serum of AOS mice. IFN-γ levels in spleen did not change but showed nonsignificant reduction in lymph nodes and significant reduction in serum in response to AOS. TNF-α levels in spleen and serum were unchanged and increased in lymph nodes. This in vivo cytokine study confirms the earlier findings that microgravity-simulated conditions induce tissue-specific immune response (Mol Cell Biochem 266: 79–85, 2004)

Simulated microgravity activates apoptosis and NF-κB in mice testis

Microgravity is known to have significant effect on all aspects of reproductive function in animal models. Recent studies have also shown that microgravity induces changes at the cellular level, including apoptosis. Our effort here was to study the effect of simulated microgravity on caspase-8 and the caspase-3 activities, the effectors of the apoptotic pathway and on the transcription factor NF-κB a signaling molecule in mouse testis. Morey-Holton hind limb suspension model was used to simulate microgravity. Caspase-8 and 3 fluorometric assays were carried out and HLS mice testis exhibited a 51% increase in caspase-8 and caspase-3 compared to the controls. A sandwich ELISA-based immunoassay was carried out for detection of NF-κB which again significantly increased in the test mice. Testosterone levels were measured using an ELISA kit and in HLS mice the decrease was significant. There was also a significant decrease in testis weight in the test mice. Simulated microgravity activates caspase 8, 3 and NF-κB necessary to stimulate the apoptotic pathway in mice testis. This may account for the drop in testis weight and testosterone level further affecting testicular physiology and function.

Gene expression profiling of human epidermal keratinocytes in simulated microgravity and recovery cultures.

Simulated microgravity (SMG) bioreactors and DNA microarray technology are powerful tools to identify “space genes” that play key roles in cellular response to microgravity. We applied these biotechnology tools to investigate SMG and post-SMG recovery effects on human epidermal keratinocytes by exposing cells to SMG for 3, 4, 9, and 10 d using the high aspect ratio vessel bioreactor followed by recovery culturing for 15, 50, and 60 d in normal gravity. As a result, we identified 162 differentially expressed genes, 32 of which were “center genes” that were most consistently affected in the time course experiments. Eleven of the center genes were from the integrated stress response pathways and were coordinately down-regulated. Another seven of the center genes, which are all metallothionein MT-I and MT-II isoforms, were coordinately up-regulated. In addition, HLA-G, a key gene in cellular immune response suppression, was found to be significantly up-regulated during the recovery phase. Overall, more than 80% of the differentially expressed genes from the shorter exposures (≤4 d) recovered in 15 d; for longer (≥9 d) exposures, more than 50 d were needed to recover to the impact level of shorter exposures. The data indicated that shorter SMG exposure duration would lead to quicker and more complete recovery from the microgravity effect.

An Environmental Assessment of Mold Concentrations and Potential Mycotoxin Exposures in the Greater Southeast Texas Area

Abstract An assessment of indoor air quality in homes from the greater Houston and Southeast Texas area was sampled over a three-month period to ascertain the potential toxic exposure to molds and mycotoxins. In 2001 Houston experienced widespread flooding due to Tropical Storm Allison. The slow receding waters allowed molds to proliferate to great levels and affected many homes and businesses in and around the Houston area. Source (air) and surface (swab, tape, and bulk) samples were taken in fungal contaminated residences to measure the amounts of airborne and surface contaminants. Indoor airborne counts of fungal spores that were identified to be 80% or more of the outside counts of similar fungal genera were classified as indoor air contaminants. This study identified species of Aspergillus, Penicillium, and Cladosporium at levels ranging from 80% to 20 times greater than the corresponding outdoor readings in the greater Southeast Texas area. Aspergillus and Penicillium species are considered potentially toxic to humans at elevated levels due to their ability to produce mycotoxins. Cladosporium is considered nonpathogenic, but can still cause allergenic reactions from hyphal fragments. Surface sampling which is utilized for identification of visible mold contaminants, identified species of Aspergillus, Penicillium, Chaetomium, Cladosporium, and Stachybotrys ranging from 1 to 100 total colony counts (1–100% for tape and bulk noncultured assays) contingent upon the extent of visual contamination. The numerical values from surface sampling were used primarily to identify the degree of visual contamination, as well as, identify any molds that may have been too heavy to become aerosolized.

The determination of sulfide in the aqueous environment

Abstract Direct measurements were made by use of a sulfide ion-selective electrode in conjunction with a double-junction reference electrode and an Orion Model 407 A/F specific ion meter. This method enables simple and rapid determinations of the total sulfide ion concentration in water. Total sulfide was determined in samples collected from lakes in Fort Bend County. Texas, which ranged from 4 ppb to 18 ppb and from the Houston Ship Channel which ranged from 13 ppb to 42 ppb. A series of experiments shows that (1) the rate of oxidation of S 2− exposed to air is about three times slower for solutions containing the sulfide antioxidant buffer (SAOB II) and (2) the rate of oxidation of S 2 in tightly capped containers is about twice as fast for solutions not containing SAOB II than for solutions containing SAOB II. It was concluded that the use of SAOB II almost eliminates the problem of air oxidation of sulfide ions during direct measurements and that sulfide ion concentrations (as low as 4 ppb) can be determined directly in the laboratory and in the field by use ofthis electrode method.

Measurement of Volatile Organic Compounds in the Urban Atmosphere of Harris County, Texas

Volatile organic compounds (VOCs) are a major component of urban air pollution. It is well documented that exposure to certain types of VOCs can cause adverse health effects such as cancer, immune and neurological damage, and reproductive and endocrine disorders. Urban air samples were collected at five locations in Harris County, Texas to determine the measurement of VOCs in the ambient air of residential areas in close proximity to industrial facilities that emit toxic air pollutants into the air. Three locations used in this study were located along the Houston Ship Channel (HSC), in the heart of one of the largest petrochemical complexes in the nation. Two other sampling locations were located many miles away from the ship channel and any industrial facilities that are required to report toxic air emissions. Air samples were collected daily over an 8-h period from December 2002 to March 2003. The samples were collected in 6-L stainless steel Silonite-coated canisters and analyzed using a modified version of EPA Method TO-15. A total of 53 compounds was quantitated using a gas chromatograph mass spectrometer system coupled to a cryogenic preconcentrator. Eighteen alkanes and oxygenated compounds were identified, along with 7 alkenes and 5 aromatic compounds. Several alkanes such as butane, isobutane, 2-methyl butane, and pentane were detected at all five sites. The total VOC concentrations determined were highest at two of the industrial sites and lowest at the site farthest away from the ship channel and any industrial facilities. This study concluded that the atmosphere near Harris Countys industrial complex had higher concentrations of VOCs than the atmosphere in areas farther away from the HSC. The atmosphere of areas downwind from emission sources were found to be directly affected by toxic air emissions from industrial process but not at the levels seen in areas closer to the HSC.