Exposure to titanium dioxide and other metallic oxide nanoparticles induces cytotoxicity on human neural cells and fibroblasts

The use of titanium dioxide (TiO₂) in various industrial applications (eg, production of paper, plastics, cosmetics, and paints) has been expanding thereby increasing the occupational and other environmental exposure of these nanoparticles to humans and other species. However, the health effects of exposure to TiO₂ nanoparticles have not been systematically assessed even though recent studies suggest that such exposure induces inflammatory responses in lung tissue and cells. Because the effects of such nanoparticles on human neural cells are unknown, we have determined the putative cytotoxic effects of these nanoparticles on human astrocytes-like astrocytoma U87 cells and compared their effects on normal human fibroblasts. We found that TiO₂ micro- and nanoparticles induced cell death on both human cell types in a concentration-related manner. We further noted that zinc oxide (ZnO) nanoparticles were the most effective, TiO₂ nanoparticles the second most effective, and magnesium oxide (MgO) nanoparticles the least effective in inducing cell death in U87 cells. The cell death mechanisms underlying the effects of TiO₂ micro- and nanoparticles on U87 cells include apoptosis, necrosis, and possibly apoptosis-like and necrosis-like cell death types. Thus, our findings may have toxicological and other pathophysiological implications on exposure of humans and other mammalian species to metallic oxide nanoparticles.     

Oxidative degradation of model lipids representative for main paper pulp lipophilic extractives by the laccase–mediator system

Different model lipids-alkanes, fatty alcohols, fatty acids, resin acids, free sterols, sterol esters, and triglycerides-were treated with Pycnoporus cinnabarinus laccase in the presence of 1-hydroxybenzotriazole as mediator, and the products were analyzed by gas chromatography. The laccase alone decreased the concentration of some unsaturated lipids. However, the most extensive lipid modification was obtained with the laccase-mediator system. Unsaturated lipids were largely oxidized and the dominant products detected were epoxy and hydroxy fatty acids from fatty acids and free and esterified 7-ketosterols and steroid ketones from sterols and sterol esters. The former compounds suggested unsaturated lipid attack via the corresponding hydroperoxides. The enzymatic reaction on sterol esters largely depended on the nature of the fatty acyl moiety, i.e., oxidation of saturated fatty acid esters started at the sterol moiety, whereas the initial attack of unsaturated fatty acid esters was produced on the fatty acid double bonds. In contrast, saturated lipids were not modified, although some of them decreased when the laccase-mediator reactions were carried out in the presence of unsaturated lipids suggesting participation of lipid peroxidation radicals. These results are discussed in the context of enzymatic control of pitch to explain the removal of lipid mixtures during laccase-mediator treatment of different pulp types.     

Influence of Bentonite Proportion in Natural Clay on Pb2+ ions Sorption: Response Surface Methodology,Kinetics and Equilibrium Studies

The influence of natural clay's bentonite proportion on Pb²⁺ sorption capacity was investigated using response surface methodology (RSM), kinetics and equilibrium studies. Experiments were conducted at different initial pH (3–7), bentonite to clay ratio (0–100%), initial Pb²⁺ ions concentration (20–120 mg/L) and sorbent dosage (0.2–1 g). Under the RSM study, data obtained from 27 experiments undertaken were found to fitted second-order polynomial model (R² = 0.998 and R²-predicted = 0.994). Analysis of variance showed that the Pb²⁺ sorption capacity was influenced according to the order; initial concentration> mass of adsorbent > initial pH > bentonite proportion. Optimal operating conditions were obtained at initial pH 5, 0.2 g sorbent dosage, 30% bentonite to clay ratio and 100 mg/L Pb²⁺ ion concentration. Regardless of the bentonite proportion, Pb²⁺ sorption kinetics followed pseudo-second-order associated with intra-particle diffusion. The sorption isotherm for the clay which was described by Freundlich yielded higher adsorption capacity (25 mg/g) while that of the bentonite was described by Langmuir model with lower maximum sorption capacity of 15 mg/g. These results suggest that sorption of the Pb²⁺ ions was not likely to significantly impact on the removal of Pb²⁺ ions during electrokinetic remediation process of clay having different proportion of bentonite.     

A Biomathematical Modeling Approach to Explain the Phenomenon of Radiation Hormesis

This study presents an improved version of a published biomathematical model, the Random Coincidence Model-Radiation Adapted (RCM-RA). That model describes how cancer mortality increases as dose rate increases in the high-dose rate range, as well as how mortality decreases as dose rate increases in the low-dose rate range. It was assumed that low-dose rates of ionizing radiation induce cellular defense mechanisms that also prevent or repair endogenous DNA damage caused by natural cell metabolism. The model presented describes the development of cancer by a phase of initiation that consists of a series of DNA lesions in the critical regions of tumor-associated genes such as proto-oncogenes or tumor-suppressor genes. Initiated cells can divide and form a clone of initiated cells. This clonal growth is called promotion and leads to premalignant cells. Premalignant clones can sustain further genomic damage that may lead to a malignant cell and ultimately a malignant tumor. The model thereby shares structural features with Moolgavkar's two-stage clonal expansion model. It was tested on published, U-shaped data of radon exposure in U.S. homes. The model correctly reflects the ratio of endogenous DNA damage to radiation-induced damage.     

Static magnetic field measurements in residences in relation to resonance hypotheses of interactions between power-frequency magnetic fields and humans.

Bowman et al. used epidemiologic data to test a model in which subjects were classified as being "in-resonance" or "not-in-resonance" for 60-Hz magnetic-field exposures depending on single static magnetic-field measurements at the centers of their bedrooms. A second paper by Swanson concluded that a single static magnetic-field measurement is insufficient to meaningfully characterize a residential environment. The main objective of this study was to investigate exposure-related questions raised by these two papers in two U.S. data sets, one containing single spot measurements of static magnetic fields at two locations in homes located in eight states, and the other repeated spot measurements (seven times during the course of one year) of the static magnetic fields at the centers of bedrooms and family rooms and on the surfaces of beds in 51 single-family homes in two metropolitan areas. Using Bowman's criterion, bedrooms were first classified as being in-resonance or not-in-resonance based on the average of repeated measurements of the static magnetic field measured on the bed where the presumed important exposure actually occurred. Bedrooms were then classified a second time using single spot measurements taken at the centers of bedrooms, centers of family rooms, or on the surfaces of beds, as would be done in the typical epidemiologic study. The kappa statistics characterizing the degree of concordance between the first (on-bed averages) and second (spot measurements) methods of assessing resonance status were 0.44, 0.33, and 0.67, respectively. This level of misclassification could significantly affect the results of studies involving the determination of resonance status.     

Temporal pattern of LH secretion: regulation by multiple ultradian oscillators versus a single circadian oscillator

The possibility that the 24h rhythm output is the composite expression of ultradian oscillators of varying periodicities was examined by assessing the effect of external continuously or pulsed (20-minute) Gonadotropinreleasing hormone (GnRH) infusions on in vitro luteinizing hormone (LH) release patterns from female mouse pituitaries during 38h study spans. Applying stepwise analyses (spectral, cosine fit, best-fit curve, and peak detection analyses) revealed the waveform shape of LH release output patterns over time is composed of several ultradian oscillations of different periods. The results further substantiated previous observations indicating the pituitary functions as an autonomous clock. The GnRH oscillator functions as a pulse generator and amplitude regulator, but it is not the oscillator that drives the ultradian LH release rhythms. At different stages of the estrus cycle, the effect of GnRH on the expression of ultradian periodicities varies, resulting in the modification of their amplitudes but not their periods. The functional output from the system of ultradian oscillators may superimpose a “circadian or infradian phenotype” on the observed secretion pattern. An “amplitude control” hypothesis is proposed: The temporal pattern of LH release is governed by several oscillators that function in conjunction with one another and are regulated by an amplitude-controlled mechanism. Simulated models show that such a mechanism results in better adaptive response to environmental requirements than does a single circadian oscillator. (Chronobiology International, 18(3), 399-412, 2001)     

Inactivation of BK channels mediated by the NH(2) terminus of the beta3b auxiliary subunit involves a two-step mechanism: possible separation of binding and blockade

A family of auxiliary beta subunits coassemble with Slo alpha subunit to form Ca(2)+-regulated, voltage-activated BK-type K(+) channels. The beta subunits play an important role in regulating the functional properties of the resulting channel protein, including apparent Ca(2)+ dependence and inactivation. The beta3b auxiliary subunit, when coexpressed with the Slo alpha subunit, results in a particularly rapid ( approximately 1 ms), but incomplete inactivation, mediated by the cytosolic NH(2) terminus of the beta3b subunit (Xia et al. 2000). Here, we evaluate whether a simple block of the open channel by the NH(2)-terminal domain accounts for the inactivation mechanism. Analysis of the onset of block, recovery from block, time-dependent changes in the shape of instantaneous current-voltage curves, and properties of deactivation tails suggest that a simple, one step blocking reaction is insufficient to explain the observed currents. Rather, blockade can be largely accounted for by a two-step blocking mechanism (C(n) <---> O(n) <---> O(*)(n) <---> I(n)) in which preblocked open states (O*(n)) precede blocked states (I(n)). The transitions between O* and I are exceedingly rapid accounting for an almost instantaneous block or unblock of open channels observed with changes in potential. However, the macroscopic current relaxations are determined primarily by slower transitions between O and O*. We propose that the O to O* transition corresponds to binding of the NH(2)-terminal inactivation domain to a receptor site. Blockade of current subsequently reflects either additional movement of the NH(2)-terminal domain into a position that hinders ion permeation or a gating transition to a closed state induced by binding of the NH(2) terminus.     

The galvanotaxis response mechanism of keratinocytes can be modeled as a proportional controller

Human keratinocytes actively crawl in vitro when plated onto a collagen-coated glass substrate, and their direction of migration is totally random. In response to an imposed dc electric field, they migrate asymmetrically, moving mostly toward the negative pole of the field. The authors have analyzed experimental data reported by others to determine the basic characteristics of the cellular response machinery in these keratinocytes. This movement can be completely described mathematically using two independent variables: the speed, V, and the angle of migration, ϕ. The authors propose a model in which a steerer (controller without feedback) is responsible for determining the speed, and an automatic controller (controller with feedback) is responsible for determining the angle of migration. The torque to rotate is induced by a deterministic cellular signal and a stochastic cellular signal. The cellular machine characteristics are determined as follows: The angular dependence of the detection unit is sin ϕ; the detection unit detects the guiding field in a linear fashion; the cellular reaction unit can be described by a constant; the chemical amplifier, as well as the cellular motor work, is linear; the cellular characteristic time, which quantifies the cellular stochastic signal, is 50 min.     

Accurate prediction of human drug toxicity: a major challenge in drug development

Over the past decades, a number of drugs have been withdrawn or have required special labeling due to adverse effects observed post-marketing. Species differences in drug toxicity in preclinical safety tests and the lack of sensitive biomarkers and nonrepresentative patient population in clinical trials are probable reasons for the failures in predicting human drug toxicity. It is proposed that toxicology should evolve from an empirical practice to an investigative discipline. Accurate prediction of human drug toxicity requires resources and time to be spent in clearly defining key toxic pathways and corresponding risk factors, which hopefully, will be compensated by the benefits of a lower percentage of clinical failure due to toxicity and a decreased frequency of market withdrawal due to unacceptable adverse drug effects.     

Apoptosis pathways in cancer and cancer therapy

Activation of apoptosis pathways is a key mechanism by which cytotoxic drugs kill tumor cells. Also immunotherapy of tumors requires an apoptosis sensitive phenotype of target cells. Defects in apoptosis signalling contribute to resistance of tumors. Activation of apoptosis signalling following treatment with cytotoxic drugs has been shown to lead to activation of the mitochondrial (intrinsic) pathway of apoptosis. In addition, signalling through the death receptor (extrinsic) pathways, contributes to sensitivity of tumor cells towards cytotoxic treatment. Both pathways converge finally at the level of activation of caspases, the effector molecules in most forms of cell death. In addition to classical apoptosis, non-apoptotic modes of cell death have recently been identified. Mechanisms to overcome apoptosis resistance include direct targeting of antiapoptotic molecules expressed in tumors as well as re-sensitization of previously resistant tumor cells by re-expression of caspases and counteracting apoptotis inhibitory molecules such as Bcl-2 and molecules of the IAP family of endogenous caspase inhibitors. Molecular insights into regulation of apoptosis and defects in apoptosis signalling in tumor cells will provide novel approaches to define sensitivity or resistance of tumor cells towards antitumor therapy and provide new targets for rational therapeutic interventions for future therapeutic strategies.This work was presented at the first Cancer Immunology and Immunotherapy Summer School, 8–13 September 2003, Ionian Village, Bartholomeio, Peloponnese, Greece.