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Nano-sized titanium dioxide (TiO2) is among the top five widely used nanomaterials for various applications. In this study, we determine the phototoxicity of TiO2 nanoparticles (nano-TiO2) with different molecular sizes and crystal forms (anatase and rutile) in human skin keratinocytes under UVA irradiation. Our results show that all nano-TiO2 particles caused phototoxicity, as determined by the MTS assay and by cell membrane damage measured by the lactate dehydrogenase (LDH) assay, both of which were UVA dose- and nano-TiO2 dose- dependent. The smaller the particle size of nano-TiO2 the higher the cell damage. The rutile form of nano-TiO2 showed less phototoxicity than anatase nano-TiO2. The level of photocytotoxicity and cell membrane damage is mainly dependent on the level of reactive oxygen species (ROS) production. Using polyunsaturated lipids in plasma membranes and human serum albumin as model targets, and employing electron spin resonance (ESR) oximetry and immuno-spin trapping as unique probing methods, we demonstrated that UVA irradiation of nano-TiO2 can induce significant cell damage, mediated by lipid and protein peroxidation. These overall results suggest that nano-TiO2 is phototoxic to human skin keratinocytes, and that this phototoxicity is mediated by ROS generated during UVA irradiation.
Nanomaterials have been recently developed that show great potential for use in the biomedical, optical, and electronic fields. They are currently manufactured on a large scale to be used in thousands of consumer goods and industrial products. However, nanomaterials are potentially toxic due to their unique properties including extremely high surface-to-volume ratios, which can make them very reactive or catalytic. Furthermore, the tiny size of nanomaterials makes it possible for them to pass through cell membranes and other biological barriers. Therefore, investigation of cellular toxicity and phototoxicity of nanomaterials is critically needed.
Nano-sized titanium dioxide (TiO2) is among the top five nanomaterials widely used in paints, plastics, cosmetics, personal care products, and as food additives and drug delivery agents (Kangwansupamonkon et al., 2009; Ray et al., 2009). The usefulness of TiO2 nanoparticles (nano-TiO2) is primarily linked to two of its functions: absorbing and deflecting ultraviolet (UV) radiation and semiconductor photocatalysis (Fujishima et al., 2000). Under UVA irradiation, electrons in the TiO2 valence band absorb the photon energy and jump to the conduction band, leaving valence band holes that extract electrons from water or hydroxyl ions and generate hydroxyl radicals (•OH). Formation of other reactive oxygen species (ROS), including superoxide (O2•−) and singlet oxygen (1O2), by different mechanisms has also been reported (Hirakawa and Hirano, 2006; Daimon and Nosaka, 2007). The photocatalytic properties make TiO2 nanoparticles successful candidates for degrading organic pollutants (Mura et al., 1999) or killing microorganisms (Kangwansupamonkon et al., 2009; Foster et al., 2011). On the other hand, potential risks associated with exposure to nano-TiO2 raise many concerns due to generation of ROS during UV irradiation; ROS are genotoxic or cytotoxic to organisms (Vevers and Jha, 2008; Ghosh et al., 2010; Sycheva et al., 2011).
Previous studies have suggested that nano-TiO2 that has not been irradiated with UV has limited cytotoxicity (Rehn et al., 2003). However, further reports have shown that TiO2 particles without illumination affected cell-matrix adhesion, which is not associated with ROS damage to cells (Fujita et al., 2009). Under UV irradiation, TiO2 nanoparticles exhibit toxic effects to both normal tissue and cancer cells, such as human dermal and lung cells (Liao et al., 2009), human keratinocyte cells (Shukla et al., 2011; Simon et al., 2011), rat lung alveolar macrophages (Afaq et al., 1998), intestinal cells (Koeneman et al., 2010) and HeLa cells (Cai et al., 1992). The human skin and eye are of particular concern as target sites for phototoxic damage because these tissues are constantly exposed to direct daily ambient radiation. It was reported that although micron-sized TiO2 cannot penetrate through the intact epidermal barrier (Kiss et al., 2008), topical application of TiO2 nanoparticles may induce dermal toxicity (Wu et al., 2009). The photoreactivity of TiO2 nanoparticles is largely dependent on their particle size, shape and crystal structure (Sharma, 2009; Liu et al., 2010). It was demonstrated that smaller nano-TiO2 particles tend to have higher phototoxicity, and anatase and amorphous forms of nano-TiO2 show higher cytotoxicity than its rutile form (Xue et al., 2010; Sanders et al., 2011).
In the present studies, we have evaluated the phototoxicity of nano-TiO2 with four different sizes (<25 nm, 31 nm, <100 nm, and 325 nm) and two crystal forms (anatase and rutile) towards human skin keratinocytes under UVA irradiation. We observe that their differences in phototoxicity are highly correlated with the production of ROS. By employing polyunsaturated lipids in plasma membrane and human serum albumin (HSA) as model targets, our unique electron spin resonance (ESR) oximetry and immuno-spin trapping techniques indicate that UVA photoirradiation of nano-TiO2 in biological systems can lead to significant cell damage.
Nano-TiO2 (A25, <25 nm anatase, catalog # 637254; A325, 325 mesh anatase, catalog # 248576; R100, <100 nm rutile, catalog # 637262), HSA, xanthine, superoxide dismutase (SOD), diethylenetriaminepentaacetic acid (DTPA), 2,2,6,6-tetramethyl-4-piperidone (TEMP) and dimethyl sulfoxide (DMSO, cell-culture grade) were all purchased from Sigma Chemical Co. (St. Louis, MO). Titanium dioxide (P25, 31 nm anatase/rutile, catalog # Aeroxide® TiO2 P25) was from Degussa (Alpharetta, GA). Fetal bovine serum (FBS) was from Biofluids (Rockville, MD, USA). Dulbecco's modified Eagle's medium (DMEM), phosphate buffered saline (PBS, pH 7.4), trypsin-EDTA, penicillin, and streptomycin were purchased from Invitrogen Inc. (Grand Island, NY). The MTS assay kit (CellTiter 96® Aqueous) was purchased from Promega Co. (Madison, WI). 15N-labeled 4-oxo-2,2,6,6-tetramethylpiperidine-d16-1-oxyl (15N-PDT) was purchased from Cambridge Isotope Labs (Andover, MA). Xanthine oxidase (XOD) was from Roche Applied Science (Indianapolis, IN). The spin-trap 5-tert-butoxycarbonyl 5-methyl-1-pyrroline N-oxide (BMPO) was purchased from Applied Bioanalytical Labs (Sarasota, FL). Egg phosphatidylcholine was obtained from Avanti Polar Lipids, Inc. (Alabaster, AL). 5,5-dimethy-1-pyrroline N-oxide (DMPO) was purchased from Dojindo laboratories (Japan).
The nano-TiO2 suspensions were prepared by first dispersing the nanoparticles in ultrapure water using a sonicator for 30 min to produce a 10 mg/ml stock nano-TiO2 suspension and then diluting according to the requirements of the experiments.
X-ray diffraction (XRD) patterns were recorded on a Philips XPert PRO MPD X-ray diffractometer (Philips, Eindhoven, The Netherlands) operated at 35 kV and 45 mA with Cu-Kα radiation. Scanning Electron Microscope (SEM) images were taken on a Hitachi S4800 field scanning electron microscope (FESEM, Japan) operating at 10 kV. Samples were suspended in ethanol solution by sonication first. Approximately 10 μl of suspensions were spin-coated on a silicon wafer at 3000 rpm for 60 seconds. Dynamic light scattering (DLS) measurements of particle size were carried out using a light scattering Zetasizer Nano-S90 light scattering instrument (Malvern Instruments, Enigma Business Park, UK).
Human HaCaT keratinocytes, a transformed epidermal human cell line (Boukamp et al., 1988), were grown to ~95% confluence in 96-well plates at 37ºC in DMEM containing 10% FBS under 95% air/5% CO2. For phototoxicity tests, cells were exposed in the dark for 4 h at 37 ºC to different TiO2s (50 and100 μg/ml) in DMEM (FBS free). Control cells were treated with DMEM alone. After incubation the medium was removed and replaced by sterile PBS containing 10 mM glucose. Cells were then irradiated with UVA from four fluorescent PUVA lamps (Houvalite F20T12BL-HO; National Biological Co. Twinsburg, OH). The maximum emission of the UVA light lamps was determined to be between 320–390 nm. The light intensities at wavelengths below 315 nm (UVB light) and above 400 nm (visible light) are approximately two orders of magnitude lower than the maximum in the 320–390 nm spectral region (Zhao et al., 2010). The UVA exposure doses were 0, 2.5, 5.0, and 10 J/cm2 respectively, as measured with an YSI-Kettering Model 65A Radiometer (Yellow Springs Instrument Co., Yellow Springs, OH). After exposure the PBS/glucose solution was removed and replaced with DMEM containing 2% FBS and the cells were kept in the incubator overnight. Aliquots (10 μl) were removed and LDH release into the medium was assayed using the Cytotox 96 kit (Promega Corp., Madison, WI) according to the manufacturer's directions. The remaining medium was removed and the cells were washed with PBS and replaced with 100 μl/well of PBS/glucose containing the MTS reagent (CellTiter 96 Aqueous Proliferation Assay; Promega Cop.). After incubation for 2 h at 37 ºC, the absorbance at 492 nm was recorded using a microplate reader (Spectrafluor Plus, Tecan US, RTP, NC).
Conventional ESR spectra were obtained with a Bruker EMX ESR Spectrometer (Billerica, MA). All spin trapping ESR measurements were carried out using the following settings for detection of the spin adduct between BMPO and O2•− (BMPO-OH): 10 mW microwave power, 100 G scan range and 1 G field modulation. All measurements were performed in replicates at ambient temperature. A light system consisting of a Xenon lamp coupled with a Schoeffel monochromator was used to generate UV light at 340 nm wavelength. All the samples for ESR measurements were freshly prepared in 100 mM phosphate buffer at pH 7.4. The buffer was treated with Chelex-100 resin for 24 h to remove traces of transition metal ions.
ESR oximetry measurement is based on the bimolecular collision of molecular oxygen with a spin label which is a stable free radical. Collision of the spin label with O2 produces a spin exchange, resulting in shorter relaxation times and ESR signals with broader linewidths and decreasing peak height. Thus, oxygen consumption by such mechanisms as lipid peroxidation results in decreased line widths and increased peak height for the spin probe. The oximetry method determines the oxygen concentration by measuring the hyperfine structural changes at the low field line of the ESR spectrum using 15N-PDT as the spin label (Halpern et al., 1994; Swartz and Clarkson, 1998; Xia et al., 2007; Yin et al., 2008; Yin et al., 2009). During lipid peroxidation, the levels of dissolved O2 vary, with a time dependent decrease in linewidth and an increase in the peak intensity of the ESR signal indicating continuous O2 consumption. The value of oxygen concentration was obtained from a calibrated curve of the ESR linewidth versus the oxygen concentration.
Liposomes were prepared as previously described (Kusumi et al., 1986). Briefly, a suspension of 30 mg/ml egg PC liposomes and 0.1mM 15N-PDT with or without TiO2 was added to a quartz capillary tube. The lipid peroxidation was initiated by UV (340 nm) irradiation. The ESR spectra were then recorded at 4 min intervals for 16 min. Signals were obtained with a 0.05 G scanning width, 0.5 mW incident microwave power and with 100 kHz field modulation. All ESR spectra were recorded at the low field line of 15N-PDT and at ambient temperature.
HSA protein samples were typically prepared by treating 600 μM HSA with 0.1 mg/ml TiO2, and 50 mM DMPO in 100 mM phosphate buffer (Chelex-treated with 25 μM DTPA) at pH 7.4 in a 6-well plate in the absence or presence of UVA irradiation (10 J/cm2). After treatment, samples were frozen at -80 ºC for further experiments.
The ELISA assay to measure DMPO nitrone adducts was performed essentially as described by Ehrenshaft and Mason (Ehrenshaft and Mason, 2006) with minor modifications. Two microgram aliquots of protein diluted in bicarbonate buffer, pH 9.4, were allowed to adsorb to the wells of a 96-well microtiter plate (Grenier Labortechnik) for 1.5 h. After washing once with wash buffer (1×PBS with 0.1% tween), the microtiter plate was incubated for 1 h in a 1:5000 dilution of rabbit polyclonal anti-DMPO antiserum, washed 4 times with wash buffer, incubated for 1 h more in a 1:5000 dilution of an alkaline phosphatase anti-rabbit secondary antibody, again washed 4 times with wash buffer, and finally washed once with 1×TBS, pH 9.6. After addition of CDP-Star (25 μM) (Roche Laboratories), the chemiluminescent product was detected using a microplate reader (Spectrafluor Plus, Tecan US, RTP, NC).
For Western Blot analysis, 10 μg of HSA protein from each reaction were electrophoresed under reducing conditions through a 4–12% BisTris NuPAGE acrylamide gel (Invitrogen) followed by electroblotting to a nitrocellulose membrane. Western Blot analysis was then performed as described (Ehrenshaft and Mason, 2006) except that DMPO-containing proteins were visualized using the Odyssey infrared imaging system (Licor).
The XRD results shown in Fig. 1A reveal the crystal structure of nano-TiO2 samples, which are consistent with the crystallinity provided by the vendors. The crystalline composition of P25 was evaluated to be around 86% anatase and 14% rutile by the ratios of the integral areas of the three main diffraction peaks of the two phases (Zhao et al., 2008). SEM was used to visualize the particle size and shape of nano-TiO2. The SEM images of four nano-TiO2 samples are shown in Fig. 1B. The sizes are roughly consistent with the sizes claimed by the vendors. From the SEM images we also find that aggregates or big clusters are formed for all samples, especially for the smallest A25 sample. DLS measurements show the size and distribution of TiO2 aggregates in the aqueous medium, which are largely dependent on the concentration, dispersant, and primary particle size (Sanders et al., 2011). As shown in Fig. 1C, aggregates of the smallest A25 sample were between 150–500 nm and the other three were between roughly 250–700 nm at the lower concentration (50 μg/mL) tested in the phototoxicity experiments.
The phototoxicity of the four different sizes of nano-TiO2 towards HaCaT keratinocytes was determined using two different assays. The MTS assay measures the activity of (mainly mitochondrial) dehydrogenases, while cell membrane damage is measured by the release of cytosolic LDH. As measured by the MTS assay, no decrease in viability was observed when HaCaT cells were exposed to TiO2 samples or to UVA alone (Fig. 2A and 2B). However, in the presence of UVA radiation, two anatase TiO2 samples (A325 and A25) and P25 TiO2 caused UVA dose- and concentration-dependent damage to HaCaT cells. The rutile TiO2 sample (R100) showed much less phototoxicity towards HaCaT cells even at a high concentration of 100 μg/ml and 10 J/cm2 UVA exposure (Fig. 2B). The extent of photocytotoxicity to cells irradiated at 7.5 or 10 J/cm2 was in the following order: P25>A25>A325>R100. A similar result was seen with the LDH release assay (Fig. 3A and 3B).
ESR spin trap spectroscopy was employed to determine whether or not nano-TiO2 under UVA light irradiation has the capability to generate ROS chemically. Generation of hydroxyl radicals was detected by the ESR spin trapping technique using BMPO. BMPO is a newly developed spin trap offering more advantages over the existing spin traps, such as greater stability and sensitivity (Zhao et al., 2001). The BMPO/•OH adduct has an ESR signal pattern similar to that of DMPO/•OH, which exhibits a characteristic set of four lines (1:2:2:1) in its ESR spectrum. In the absence of UVA irradiation, the ESR signal of the hydroxyl radical spin adduct was not detectable from the mixture of BMPO and nano-TiO2 (data not shown). No such ESR profile was generated when BMPO was irradiated under UVA light (340 nm) in the absence of nano-TiO2 (Fig. 4A) or in the presence of the rutile R100 TiO2 sample (Fig. 4B). However, UVA irradiation of BMPO in the presence of A325 nano-TiO2, A25 nano-TiO2, or P25 nano-TiO2 all produced the characteristic ESR profile (Fig. 4C, 4D, and 4E). As shown in Fig. 4, UVA irradiation of P25 nano-TiO2 resulted in the strongest signal of BMPO/•OH signal (Fig. 4E), followed by two anatase TiO2 samples, A325 nano-TiO2 and A25 nano-TiO2 (Fig. 4C and 4D). This signal was largely inhibited by the addition of 20% DMSO (Fig. 4F), further confirming the generation of OH radicals during the UVA irradiation.
We tested the generation of singlet oxygen by the ESR trapping technique using TEMP. It has been previously reported that 4-oxo-2,2,6,6-tetramethylpiperidine-N-oxyl (TEMPONE), a nitroxide radical detectable by EPR spectra, is generated from TEMP and singlet oxygen (Eq. 1) (Lion et al., 1976).
Irradiation of different nano-TiO2 samples (0.1 mg/ml) containing 20 mM TEMP resulted in ESR spectra of three lines with equal intensities (aN = 16.0 G), typical of nitroxide radicals (Fig. 5, inset). The hyperfine splitting constant and g factor of the photosensitized oxidation product of TEMP were identical to those of commercial TEMPONE. Fig. 5 shows the ESR intensity of the TEMPONE signal as a function of time during irradiation of different nano-TiO2 samples. The P25 nano-TiO2 solution (Fig. 5, curve A) showed the rate that increased the fastest, followed by the A25 nano-TiO2 sample (Fig. 5, curve B). The rates of production of singlet oxygen were much slower for A325 nano-TiO2 and R100 nano-TiO2 (Fig. 5, curves C and D). A control solution with TEMP only (data not shown) indicated no ESR signal increase. For P25 nano-TiO2, addition of a singlet oxygen quencher, NaN3, caused a substantial decrease in the rate of increase of the EPR signal (Fig. 5, curve E).
It has been reported that O2•− can be oxidized to 1O2 during TiO2 photocatalytic reactions (Lipovsky et al., 2012). Superoxide (O2•−) undergoes dismutation to produce H2O2, from which the •OH radical can also be formed through the Fenton reaction. To further elucidate the mechanisms of •OH and 1O2 generation, we studied the effect of SOD on the production of •OH and 1O2 by P25 nano-TiO2 during UVA irradiation. As a control study, addition of 1.25 U/ml SOD significantly inhibited the production of O2•− generated in a Xanthine system (Fig. 6A and 6B). However, no obvious effect was observed on •OH generation by P25 nano-TiO2 under UVA irradiation even at a high concentration of SOD (6 U/ml, Fig. 6C and 6D), indicating that nano-TiO2-generated •OH radicals were not evolved from O2• , at least for the most part. In contrast, the 1O2 generation was markedly reduced in the presence of 1.25 U/ml SOD (Fig. 6E and 6F), confirming that O2•− may be a main pathway in UVA-induced 1O2 generation by nano-TiO2 (Lipovsky et al., 2012). It is noteworthy that although O2•− was formed from UVA irradiation of nano-TiO2 and was attributed to the formation of 1O2, no obvious O2•− production was detected in our ESR study using BMPO as a trapping agent (Fig. 4), which is actually consistent with a previous report (Konaka et al., 2001). Its short lifetime under our experimental conditions warrants further investigation.
It is well known that ROS can produce a time-dependent peroxidation of the polyunsaturated lipids in plasma membrane. In this study, lipid peroxidation produced by UVA irradiation of different nano-TiO2 was assessed using ESR oximetry recorded at 4-min intervals for 16 min. The consumption of oxygen accompanying lipid peroxidation was measured as a time-dependent narrowing of the ESR signal for the spin probe 15N-PDT. As shown in Fig. 7, narrowing of the ESR signal was necessarily accompanied by an increase in the peak height of the ESR signal within the scan range. The results of lipid peroxidation produced by different nano-TiO2 samples (R100, A325, A25, and P25) after UVA irradiation are shown in Fig. 7B–7E, respectively. The final ESR signal intensities of R100 nano-TiO2 (Fig. 7B), A325 nano-TiO2 (Fig. 7C), A25 nano-TiO2 (Fig. 7D), and P25 nano-TiO2 (Fig. 7E) were approximately 7.5, 11.7, 16.2, and 26.3% higher than in the control (Fig. 7A). The progressive increases in peak-to-peak signal intensity (and accompanying progressive narrowing of line width) in each panel were due to time-dependent oxygen consumption resulting from lipid peroxidation. Fig. 6F presents the data as a decrease in oxygen concentration, reflecting the variations in the slope (oxygen concentration versus time) with different nano-TiO2, which decreases in the order of P25 > A25 > A325 > R100.
To further characterize the ability of ROS to oxidize target proteins, the most abundant plasma protein (human serum albumin, HSA) was incubated with DMPO and different nano-TiO2 samples with and without UVA irradiation. The reaction products were analyzed by ELISA and Western blotting using an anti-DMPO polyclonal antibody. Immunochemical detection of HSA-DMPO nitrone adducts was performed according to our previous studies (Ranguelova et al., 2010). As measured by both ELISA and Western blotting, HSA with DMPO exposed to different nano-TiO2 samples or UVA alone produced nitrone adducts at background levels (Fig. 8A–8E). While in the presence of UVA irradiation, P25 nano-TiO2 resulted in a significant increase in HSA-DMPO-derived nitrone adducts (Fig. 8I). The other three nano-TiO2 samples caused modest increases in adduct formation (Fig. 8F–8H). The order of the extent of nitrone adducts is the same as for photocytotoxicity, hydroxyl radical, singlet oxygen, and lipid peroxidation measurements.
TiO2 is regarded as an inert and non-toxic substance by many regulatory bodies; e.g., the FDA approved TiO2 as a food color additive in 1996, and the Material Safety Data Sheets (MSDS) indicate that there are no exposure hazards to the health of occupational workers and public health. However, there have been several studies that have shown that TiO2 is a potential carcinogen and photocatalyst in biological tissues (Burnett and Wang, 2011). The cytotoxicity and/or phototoxicity of nano-TiO2 is still debated. Also, there is uncertainty about whether the properties of nano-TiO2 differ with its particular form and size. Human skin and ocular tissues are most affected by ambient radiation. In vitro studies have shown that when irradiated, the smallest form of TiO2, anatase (<25 nm), is phototoxic to human retinal cells (Sanders et al., 2011). UV-induced production of ROS and the resultant oxidative stress exposure plays an important role in photocarcinogenesis caused by UV irradiation. ROS are believed to be involved in many inflammatory skin disorders, skin cancer formation, and skin aging (He et al., 2005). Thus we report here a study of the photocytotoxicity and phototoxicity of nano-TiO2 towards human HaCaT keratinocytes.
Our results show that all nano-TiO2 tested in this paper caused UVA and nano-TiO2 dose-dependent phototoxicity in HaCaT cells. As expected, smaller particles resulted in higher phototoxicity than larger particles. Rutile nano-TiO2 showed less phototoxicity than anatase nano-TiO2. Moreover, small mixed anatase/rutile nano-TiO2 (P25) showed the highest phototoxicity, which is consistent with previous reported results (Hurum et al., 2003). The cellular toxicity of nano-TiO2 mainly depends on its ROS generation under UVA irradiation. Using ESR and spin trapping technique, a "gold standard" and state-of-the-art tool for detecting and quantifying ROS, we confirmed the generation of hydroxyl radicals and singlet oxygen. ROS generation by these four nano-TiO2 particles is consistent with the phototoxicity studies, further confirming our hypothesis that ROS production was probably involved in the phototoxic mechanism.
In addition to measurement of ROS generation with ESR, we explored two other assays to further characterize the phototoxicity of nano-TiO2. ROS are known to produce a time-dependent peroxidation of the polyunsaturated lipids in plasma membrane (Yin et al., 2008; Yin et al., 2009) as well as peroxidation of lipids and proteins inside the cells (Girotti, 1998). To mimic oxidation reactions with substrates in biological systems, we used polyunsaturated lipids in plasma membranes and human serum albumin as model targets for ROS. Using innovative ESR oximetry and immune-spin trapping techniques, we easily detected ROS-induced peroxidation, which was found to be in agreement with the ESR-detected ROS production. These results indicate that both methods are useful for qualitative screening of the phototoxicity of nano-TiO2.
Based on the results reported in this study, we propose the mechanism shown in Fig. 9. UVA irradiation results in electron-hole pairs in nano-TiO2. The photoexcited nano-TiO2 serves as the initiating species to transfer energy to molecular oxygen and generate singlet oxygen (1O2) or to extract electrons from water or hydroxyl ions to generate hydroxyl radicals (•OH). Our experimental results also indicate that part of the 1O2 formation proceeds via a superoxide-dependent mechanism, whereas the OH formation is not via superoxide (Fig. 6). It is well established that in the presence of a lipid or protein, the generated ROS can initiate lipid and/or protein peroxidation. Both ROS and lipid peroxidation are associated with aging-related diseases, including cancer (Aust et al., 1993; Xia et al., 2007).
In conclusion, the studies presented here give an overall in vitro assessment of the phototoxicity by various nano-TiO2 particles towards HaCaT cells under UVA irradiation. This phototoxic damage was apparently mediated by production of ROS by photo-activated nano-TiO2. According to our results ROS are generated by UVA irradiation of nano-TiO2 with anatase and/or mixed anatase/rutile forms. Moreover, the phototoxicity of nano-TiO2 was less with larger particle size and surface areas, indicating that coarse particles of nano-TiO2 may consistently produce less ROS.
This article is not an official US Food and Drug Administration (FDA) guidance or policy statement. No official support or endorsement by the US FDA is intended or should be inferred. This work was supported by a regulatory science grant under the FY11 FDA Nanotechnology CORES Program (JJ Yin) and the Intramural Research Program of the NIH, National Institute of Environmental Health Sciences. The authors are indebted to Dr. Jianxun Xu for his help in SEM image analysis, Dr. Zhanjun Gu, Yeteng Zhong for their help in XRD analysis, and Dr. Ann Motten, NIEHS, for critical reading of the manuscript.
Conflict of interest statement
The authors declare that there are no conflicts of interest.
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Rocsi Diaz is wasting no time in stripping down into a piece bathing suit to reveal her incredible figure during her holiday vacay with alleged friend/
What do you think of Rocsi and Eddie's secret romance?
Cress arugula peanut tigernut wattle seed kombu parsnip. Lotus root mung bean arugula tigernut horseradish endive yarrow gourd. Radicchio cress avocado garlic quandong collard greens.
So how the hell y'all know she was there with him and his family if we only saw pics of her?!? Come on GWL step ya game up!ReplyDelete
The second to last pic is Eddie's daughter and GWL did do a story a few days ago showing eddie and rocsi chilling from their balcony. Plus alot of other media outlets reported the same story too.
Plus CINDY what happened to Shane aka Gossipwelove writer?? I havent read a story from him in a couple of days now.
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Apollo Group (APOL) reported adjusted earnings (excluding restructuring and other charges) of 58 cents per share for the second quarter of 2012, which nicely beat the Zacks Consensus Estimate of 39 cents. However, quarterly earnings were down approximately 30% from the prior-period earnings of 83 cents primarily due to lower revenues. Apollo delivered total revenue of $969.6 million during the quarter, down 7.5% from the year-ago quarter due to an 8% decline in revenues from the University of Phoenix, the company's wholly owned subsidiary. However, total revenue breezed past the Zacks Consensus Estimate of $933 million.Quarter in Detail Revenues declined at the University of Phoenix in the reported quarter due to a 12.2% decrease in degreed enrollment to 355,800 students. New enrollment growth (also called new degreed enrollment growth) was up only 1% from the prior-year period, a disappointment from management's original expectation of maintaining the sharp 13% rise witnessed in the sequentially preceding quarter. In late February 2012, management had however tempered its growth expectation to the low single digits. Sluggish new enrollment growth was affected by company specific factors, an improving job market, robust competition and a volatile economy. Higher education sees a countercyclical movement during weaker economic conditions. Post-secondary enrollments tend to rise as both unemployed and employed workers return to school to re-skill themselves and existing students remain in school for a longer duration compelled by the intensely competitive job market. Enrollments are once again taking a beating with an improving job market. Instructional and student advisory cost inched up 1.9% to $429.5 million during the quarter. As a percentage of net revenue, cost grew 410 basis points (bps) primarily due to the company's continuous effort to improve its services (especially technology) in order to enhance students' educational outcomes. Marketing expenses, incurred to establish relationships with employers and community colleges, rose 1.3% to $159.3 million. Admission advisory expenses declined 0.9% to $101.4 million compared with the prior-year figure, but increased 60 bps as a percentage of net revenue. Benefits from lower admissions advisory headcounts were offset by increased employee compensation costs. Though Apollo's general and administrative (G&A) expenses declined 0.4% to $84.0 million, as a percentage of net revenue G&A shot up by 70 bps. Bad debt expenses declined 110 bps as a percentage of revenues to 3.2% due to lower enrollments.Balance Sheet Apollo ended the quarter with cash and cash equivalents of $800.7 million compared with $1.20 billion at the end of the first quarter of 2012 due to share repurchases in the current quarter. During the reported quarter, Apollo repurchased approximately 6.4 million shares worth $328.8 million. Moreover, subsequent to quarter end, on February 29, the company made an additional expenditure of $92.5 million toward repurchasing 2.2 million shares. The company is thus left with $0.5 million under its current $500 million share repurchase authorization. At the end of the quarter, the company's total outstanding debt (including short and long term) stood at $127.5 million compared with $114.3 million at the end of the first quarter of 2012.Fiscal 2012 Outlook At the second quarter conference call, the company maintained its previously provided revenue guidance of $4.1-$4.3 billion and adjusted operating income guidance of $625-$725 million for fiscal 2012. We would like to remind investors that in late February 2012, the company had reduced its adjusted operating income guidance from $655-$750 million to $625-$725 million. The cut in the guidance was in anticipation of fluctuating new degreed enrollment. The company expects new degreed enrollment growth to remain volatile through the rest of 2012. If the current trends continue the third quarter could witness double-digit declines in new degreed enrollment.Long-Term Initiatives The company is focusing on corporate partnerships as it sees opportunity for growth in this area. It is reallocating marketing and advertising costs to foreground its innovative pipeline. To its credit, Apollo is also working to cut costs and to provide world class affordable education to its students with a focus on building long-term. Management believes these initiatives will have a substantial impact on the cost structure beginning in fiscal 2013 with an even greater impact in fiscal 2014. APOLLO GROUP (APOL): Free Stock Analysis Report To read this article on Zacks.com click here.
updated Mar 27, 2012
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Programs developed by ScenicReflections.com
Animated screensaver featuring a scene of a water stream falling over a group of rocks and into a pond. The animation is less than perfect and matches poorly with the realistic photographic background. The installation wizard suggests you to install additional software, but you can refuse to do so.
The free Christmas Scenic Reflections Screensaver displays 80 magnificent Christmas themed images accompanied by nearly 60 minutes of beautiful, cd-quality Christmas music. This free Christmas screensaver is perfect to help get you into the Christmas Spirit!This free Christmas screensaver features six beautiful, full-length, CD-quality musical compositions.
3D Christmas Fireplace will bring warmth to you and your family. This beautiful screensaver will allow you, and me of course, to enjoy one of the nicest scenes of the Christmas season. You will be able to sit down on your favorite couch and just relax and enjoy watching a beautifully decorated living room. You will see a very beautiful, and of course, decorated fireplace.
It is a beautiful animated desktop customizable screensaver. You will be enchanted by this picturesque landscape: warm summer day, small bay right by the sea shore, exotic nature, huge rocks, cascade of falling water, smooth sea, butterflies fluttering from the flowers, natural growth, and birds flying in the sky.
Beautiful screensaver featuring three different aquarium settings with sound and six species of tropical fish. It also features a cool-looking clock showing the current time. You can adjust many of the screensaver's settings such as the sound, the number of fish you want to display (up to 30), and so on. If you love fish, this is the right screensaver for you.
The Wild West Screensaver is a simple screensaver that displays images of the typical wild west. The screensaver includes images displaying cowboys on horses in desert landscapes and beautiful sunsets. The screensaver also shows a bar that slides vertically and horizontally over the included images highlighting the colors in that part.
Christmas Cottage Screensaver is a screensaver with a Christmas theme that shows a cute cottage with snowed roofs and snow falling. There is also a nice snowman at the doorstep. The image is mainly static, but you will see a few birds flying around and Christmas lights turning on and off every second all around the cottage.
Winter Wonders is a screensaver that features a slideshow of photographs with beautiful landscapes of the world in winter time, and their snowed mountains and valleys, frozen lakes, pine forests and mirrored waters. Winter Wonders is absolutely free to download and use, and is a good choice for any screensaver collection.
The free Snowy Hut 3D Animated Screensaver by Scenic Reflections shows snow as it slowly descends upon the ground and a country hut. Watch the time slowly pass by in real time! As your day progresses into night, so does the screensaver, until a beautiful full moon shines through the night sky.
Hot Rod Cars is a cool screensaver that will show you 75 high-resolution images of the most incredible classic cars and street rods. You will enjoy viewing different American car syles, apart from hot rod or street rod style, you will see street machine style, low rider, drag racing and many others dating from the 30's, 40' and 50's.
Spider Web Screensaver is an animated desktop application that shows a spider web that weaves until completely covering your screen. The animation is very simple, but well made and attractive, and together with the ghostly sounds and horror voices, makes a perfect combination that can really chill your blood.
Winter Valley 3D Screensaver is a nice screensaver that shows a snowy valley with birds flying around and snow falling from the sky. The image is somewhat cartoonish and is pretty well created, looking almost like a real landscape. You will be able to see some trees, a fence, snowy mountains in the distance, a woodpecker on a tree, and different birds that come and go all the time.
The Free Pedigree Dogs Scenic Reflections Screensaver displays 85 images of some of the most adorable, precious canines you may ever see. You're going to be sitting pretty with this screensaver, that's for sure. This free dog screensaver also features three beautiful, full-length, CD-quality musical compositions.
Thanksgiving Scenic Reflections Screensaver displays 50 wonderful Thanksgiving themed images accompanied by over 20 minutes of beautiful, cd-quality music. This free Thanksgiving screensaver is perfect to help get you into the holiday mood! It features 7 beautiful, full-length, CD-quality musical compositions.
Now you can get all the affection from this canine companion with none of the slobber! The free Golden Retriever Pup Dog Licking Screen Cleaner Screensaver includes nearly 20 minutes of beautiful, CD quality piano music that is perfect for the true dog lover, and now you can optionally add your own music tracks to help customize the screensaver even further.
This screensaver, developed by Scenic Reflections, allows you to observe more than seventy photographs of the sky and the space. Air and Space Scenic Reflections Screensaver displays 76 images of some of the most awesome Air and Space themed images.
Dinosaur Habitat Screensaver takes you on a journey to the past. This awesome screensaver will immediately transport you to the typical scene of that era. Large open plains, a couple of volcanoes, a swamp or a lake, and dinosaurs. And it will not be a still scene with the image of frozen animals. This screensaver is completely animated.
The free Halloween Scenic Reflections screensaver is the first Halloween themed slideshow screensaver to date. But just because it`s a slideshow style screensaver, don`t think it won`t make your skin crawl! It displays 70 stunning Halloween themed images for your enjoyment.
A free, 3D animated screensaver featuring the image of a watermill in the middle of a forest. It features a nice selection of music pieces that add a soothing touch. Also, you can set up many of the screensaver's features. One disadvantage is that it installs the Ask toolbar and changes the startup page of your browser. If you don't mind these inconveniences, then you may like this program.
The precious, free Golden Labrador Dog Licking Screen Cleaner Screensaver features an adorable Golden Labrador that loves to please as he sends pooch smooches through your screen! Let this cute dog lick your screen clean with this wonderful screensaver. Now you can get all the affection from this canine companion with none of the slobber!


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Hybrid Physician Assistant Masters PA Programs combine the best of an online education with the experience of the college campus in a low residency format, so there is both flexibility to study around your own schedule as well as the social networking of being on campus. If this format of higher education sounds appealing, begin a search for the best Hybrid PA masters programs by keying in a preferred location, whether by city, state or country, to see where they are offered. You can also refine your search for Hybrid Physician Assistant Masters PA Programs by browsing through degree options, such as a Hybrid Master Physician Assistant Studies.
For the student who is seeking a higher education in the health care industry, hybrid physician's assistant masters programs can open up different career options within this broad profession. A physician's assistant is a healthcare professional who is licensed to practice medicine as part of a team with physicians and other providers. These qualified individuals are concerned with preventing and treating human injury and illness by providing a variety of health care services under guidance of a surgeon or physician.
Find what could be the best grad schools offering Hybrid PA programs for you. GradSchools.com offers many accredited grad schools to choose from!


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by Suzanne TitkemeyerWhen the news of the unwanted sexual encounters between Vision Forum's Doug Phillips and Lordes Torres came out followed by the closing of both Vision Forum Ministries and their for profit Vision Forum Inc it left quite a void in the world of Christian Dominionism, Patriarchy and Quiverfull movements. Everyone speculated who would take over from the powerhouse dominionist ministry of Vision Forum. But those that might have stepped in to fill the void left by the closing of Vision Forum had their own scandals to deal with: first Bill Gothard of IBLP/ATI, who is huge in the world of homeschooling and dominionism, was accused of groping teenagers working at ATI … [Read more...]
Quoting Quiverfull: God's Slave?
by Nancy Campbell from Above Rubies - A Love SlaveEditor's note: Nancy seems to think that being a wife and mother is a sort of slavery instead of an expression of free will and love in our lives. How does that fit with her idea that being a mother and wife is the highest honor and only role for a woman? … [Read more...]
Quoting Quiverfull: Nancy Steals Hanukkah Day 3 (With Some Body Shaming)?
by Nancy Campbell from Above Rubies - Family Dedication ServiceNotice at the very end, after Nancy has explain all about desecrating, cleansing and rededicating that she mentions having the children rededicate their bodies 'body temple' because of things that have come in to dirty and spoil the temple. What could a child have done that would make that even a possibility? Oh, that's right, Nancy believes that every moment of thought is sin. … [Read more...]
Quoting Quiverfull: Stealing Hanukkah Day 1 (Glomming on to Jewish Traditions)?
by Nancy Campbell from Above Rubies - My Family and I Will Never Give UpI guess Nancy isn't content to misinterpret the Holy Bible and has to horn in on Jewish holidays, twisting that to be another one of her 'traditions' that she can misunderstand. … [Read more...]
Quoting Quiverfull: Don't Complain Ever Because God?
by Nancy Campbell of Above Rubies - You Are Destined For ThisIf you're even thinking about complaining about anything Nancy Campbell, Evangelical Cultural Enforcer, is poised above your head to hit you with a Biblical fly swatter and a reminder to shut up because it could be worse. Or maybe God had planned all this torment for you to 'refine' you or as a test. What type of all loving all knowing deity tortures people for no good reason? Because Bible! … [Read more...]
Quoting Quiverfull: Isn't It Cruel To Make Barren Women Crave Babies?
by Nancy Campbell from Above Rubies - All Women Are MothersInteresting that Nancy "I Abandoned My Liberian Adoptees" Campbell is putting every women in the world in the exact same box marked 'Motherhood' and invoking a Catholic icon to do it. Most of the time it seems like the only time the Catholics are mentioned in CPM is when someone wants to toss rocks at them and say they aren't 'real Christians' … [Read more...]
Quoting Quiverfull: God Notices When No One Else Does?
by Roy Lessin as quoted by Nancy Campbell at Above Rubies - God Is NoticingThis strikes a similar theme as Kaleesha Williams recent 'Appreciation Rant' posted on Thursday Nov. 7th, 2014. Except that Nancy is attempting to enforce the thought process that you shouldn't ever expect anyone to notice or appreciate what you do, making it okay for no one in your life to say 'thanks'. … [Read more...]
Quoting Quiverfull: Do Not Be Tolerant?
by Nancy Campbell of Above Rubies - Teach The DifferenceLooks like Nancy got some push back from her own readers and didn't like it very much. Not as much fun when your readers don't mirror back your own ideas and seek to flatter you, now is it Ms. Campbell? … [Read more...]


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1 Review
This was so good, a little spicy from the rotel tomatoes and a hint of sweet from (I'm guesting) the sour cream and milk. I used tri-colored pasta and baked 10 minutes longer. The top came out a nice light golden brown and crunchy. Don't skip step 6 it really adds to the casserole. Made for *PAC Fall 2007* game


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Pay Your Tuition
If you'd like to make a tuition payment, follow these steps:
You can make a payment using cash, check, credit card, or by wire transfer. Please note that tuition is due the 1st day of class each semester.
Pay Online: Use Enterprise
When tuition is billed to your student account, you will be emailed with a personalized login link to setup your Enterprise account. Please contact the Cashier's Office to make any payments before your Enterprise account is set up.
Note to all incoming MIIS students: Your statement will not reflect any tuition charges until after you register, although you may see your pending financial aid.
Pay by Check:
Please make checks payable to:
Middlebury Institute of International Studies (MIIS)
ATTN: Cashier
460 Pierce Street
Monterey, CA 93940
Please note your Banner ID number if you have one.
Pay by Phone:
You may pay by VISA, Mastercard, Discover, or American Express by phone: Cashier at 831-647-6434.
Pay at the Cashier's Office:
You may pay in-person at the Cashier's Office located at 449 Pierce Street, Monterey, CA 93940. Our hours are Monday to Friday from 9:00 a.m. to 1:00 p.m.
Pay by Wire Transfer:
If you'd like to pay by wire transfer, please email the Cashier's Office at firstname.lastname@example.com for instructions.


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text:
Internet officially makes life harder for primary school parents
The Internet is pretty much accepted as being very, very useful. Downtime clearly agrees. But while dropping kids off at school one Downtimer was horrified to see the clever tricks that parents are using to make their kids popular. Apparently if you sign into the school portal you can see who your child's class mates are.
Your exclusive guide to CIO trends
A collection of our most popular articles for IT leaders from the first few months of 2016, including: - Corporate giants recruit digitally-minded outsiders to drive transformation - Analytics platforms to drive strategy in 2016 - Next generation: The changing role of IT leaders.
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Parents that are ahead of the game are using this to make sure their child gives a card to every child in the class. Miss out a popular kid and you are history. This Downtimer has had to re-think his strategy. It seemed great when his child claimed to hate more classmates than she liked, but it now seems you have to give everyone a card.
Bah humbug!
Good news is IT can always fail and the latest news is the school portal is down. So it's a level giving field.


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The iPhone can be used for many more things than just making calls. There are the other obvious uses like email, music and web browsing. The iPhone 4S and 5 also have a really great camera built-in for taking pictures, but the camera also comes with a flash for pictures in low light. This flash certainly has more uses than just night photos.
The iPhone flash can also be used as a flashlight. While the main display can be used to see in the dark, the camera flash is much brighter. It may not be as bright as many flashlights, but it is built into the iPhone which is something that people always carry around with them. To use the camera's flash, an app must be downloaded from the App Store. There are many free flashlight apps and some apps that cost a few dollars. After testing many options, one came out a clear winner.
Flashlight by Rik app turns the flash into a flashlight faster than any other app tested. It remembers the last used state so users can have the flashlight turn on automatically speeding the process. The only features are to turn the flashlight on or off and a strobe option. A flashlight app really does not need any other features than these, but it does need to be fast. Most of the time, when the flashlight is needed, it is needed quickly and that is the best feature for this app. Best of all, it is free.
Flashlight by Rik one of the best flashlight apps in the App Store. I use this app almost every night and it has been on my home screen for over two years. If you are looking for the greatest fast flashlight app with a simple interface, look no further than Flashlight by Rik.
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- EXCLUSIVE OFFER: Jim Cramer's Protégé, Dave Peltier, only buys Stocks Under $10 that he thinks could potentially double. See what he's trading today with a 14-day FREE pass.
- You can view the full AT&T Ratings Report.
Here's how to trade the top five names in tech.
In this weeks blogs Cramer talks about Amazon should run a stock market, the profits one can get looking down the road and how American Airlines is being victimized.
AT&T (T) is reportedly considering making an investment in Argentina as the country rolls out new telecom reforms allowing phone companies to expand into paid television services.
Charter and Comcast are the latest cable operators to try to crack the wireless market, a trend that's been going on for more than two decades.


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Enjoy barbeque immersed in the celebrated tradition of the South with flavors of world-famous BBQ styles from Tennessee, Kentucky, Texas, the Carolinas and Missouri. Offering family style dining in a rustic and casual restaurant, enticing rubs and marinades slowly smoked over hickory, oak and fruit woods, Virgil's Real BBQ will not disappoint. BBQ, home style sides, sweet ending desserts and live music on the weekend make Virgil's Southern Hospitality complete!
Hours of Operation:
Wednesdays, Thursdays and Sundays, 4pm-9pm
Fridays and Saturdays, 4pm-2am
Fridays and Saturdays Live Music, 10pm-1am


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Three Sisters Equestrian has many fine English and Western horses for lease. Full or 1/2 leases available. Call Connie Wilson at 770-605-5276 for more details.
Three Sisters Equestrian located in Cumming Ga, is a full service horseback riding lesson, horse training and boarding stable.
We offer horseback riding lessons, specializing in Hunt Seat, Western Pleasure and Trail. Riding lessons, training, showing and sales are provided by Connie Wilson of Connie Wilson Performance Horses.... (read more)
Three Sisters Equestrian is a Horseback Riding Lesson, full service boarding and training stable, conveniently located in Forsyth County near downtown Cumming. We offer riding lessons, specializing in Hunt Seat, Western Pleasure and Trail. Lessons, training, showing and sales are offered by Connie Wilson of Connie Wilson Performance Horses. Our custom 16 stall show barn includes feeding, daily turnouts on lush private 18 acres of multiple cross fenced turnout paddocks.... (read more)
Avoid Scams and Fraud - Potential areas of fraud: wire transfer, moneygrams, money orders, cashier checks, shipping, escrow, "transaction protection", "guarantee". Check out the Buyer Safety section for more info.


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All hotels in Victoria, British Columbia, Canada
1 room, 2 adults
206 Portsmouth Drive, Victoria, BC, V9C 1R9, Canada, 800-491-6126
Birds of a Feather Oceanfront Bed & Breakfast, Victoria
Lowest price$104
available on 10/05/16
Best Price Guarantee
3.5-star B&B, convenient to Fisgard Lighthouse
- Free full breakfast, free WiFi, and free parking
Main amenities
- 3 guestrooms
- Free use of bicycles
- Terrace
- Garden
- Tour/ticket assistance
- Boat tours
- Kayaking
Feel at home
- Private bathroom
- Garden
- Terrace
- Iron/ironing board
- Hair dryer
- Coffee/tea maker
What's around
- Esquimalt Lagoon Migratory Bird Sanctuary (2.1 mi / 3.4 km)
- Havenwood Park (2.5 mi / 4.1 km)
- Latoria Creek Park (2.5 mi / 4.1 km)
- Fort Rodd Hill National Historic Park (3.1 mi / 5 km)
- Fisgard Lighthouse (2.9 mi / 4.6 km)
I spent a week at Birds of a Feather. The location is very idyllic. Once the day's…
What a pleasant surprise for a last minute trip booking to find myself at this Bed and…
TripAdvisor Traveller Rating: 4.5
From 177 reviews
- Victoria, BC (YYJ-Victoria Intl.) 36-min drive; 21.7 miles/34.9 km
- Port Angeles, WA (CLM-William R. Fairchild Intl.) 143-min drive; 38.5 miles/61.9 km
- Friday Harbor, WA (FBS-Friday Harbor Sea Plane Base) 143-min drive; 44.6 miles/71.7 km
- Friday Harbor, WA (FRD) 149-min drive; 46 miles/74.1 km
- Bedwell Harbour, BC (YBW-Bedwell Harbour Seaplane Base) 160-min drive; 44.3 miles/71.4 km
- Free self parking
Birds of a Feather Oceanfront Bed & Breakfast, Victoria
Hotel Info: 866-539-8117from $104
- King Bed Ground Floor Ensuite Room
- Luxury Waterfront Suite
- Double Bed Ground Floor Cozy Ensuite Room
- Family Waterfront Suite
- Honeymoon Suite
- Queen Bed Ground Floor Ensuite Room
More great choices
At a glance
Key facts
Hotel size
- This hotel has 3 rooms
- This hotel is arranged over 2 floors
- Check-in time 1:30 PM-midnight
- Check-out time is 11 AM
Required at check in
- Credit card or cash deposit required
- Government-issued photo ID required
Travelling with others
- No cribs (infant beds)
- Pets not allowed
- Free WiFi in public areas
- Free WiFi in rooms
- Free self parking
In the hotel
Food and drink
- Free full breakfast
Things to do
- Free bicycles on site
- Kayaking on site
- Hiking/biking trails nearby
- Tours/ticket assistance
- Garden
- Terrace
In the room
Home comforts
- Coffee/tea maker
- Bathrobes
- Iron/ironing board
Freshen up
- Private bathroom
- Hair dryer
Stay connected
- Free WiFi
Also popular with travelers from the USA
Recent reviews
Outstanding 4.7 /5 from 26 reviews
Birds of a Feather Oceanfront Bed & Breakfast, Victoria
Perfect get away
I went on a trip to Vancouver Island with my Dad and was looking for something a little different. This quiet and beautiful location on a bird sanctuary was perfect. I also found it difficult to find a B&B with more than one bed, which is what we needed. Great hospitality, superb filling home cooked breakfast. We really enjoyed the other guests we met at this spot too. I would definitely return when visiting the Victoria area.
Birds of a Feather Oceanfront Bed & Breakfast, Victoria
Peace and quiet outside the city
We had a fantastic stay just outside of Victoria. Peace and quite outside the city with views of the harbor and light house. Will be booking this place again when we visit Victoria.
Birds of a Feather Oceanfront Bed & Breakfast, Victoria
Fabulous getaway
Lovely, restful, beautiful place. Knowledgeable and entertaining owners and staff, incredible breakfasts, well-run and overall a great place to hang out or use as a base to explore the area.
Birds of a Feather Oceanfront Bed & Breakfast, Victoria
WOW - this place is great!
This is a gorgeous place with excellent hosts! The grounds are wonderful with flowers on the decks and beautiful bay views. After hours of travel, we felt a wonderful sense of relaxation come over us upon arrival. We had some issues with our phones and Dieter was amazing in helping us take care of things in the states that were time sensitive. He really went above and beyond to help! The room was spacious and spotless. Our room had a loft with a bed for our son which he totally loved. There was a wonderful deck where we could watch the geese and swans on the bay. Our teenage son took a kayak out for a spin to work out the travel kinks. In the morning, we went for a long walk on the nearby trails and were treated to some wonderful scenery. Breakfast was excellent both days we stayed and Dieter showed us on the map the nearby points of interest. We will definitely return on our next trip to Victoria!
Birds of a Feather Oceanfront Bed & Breakfast, Victoria
Amazing trip to watch my friends get married. Walking distance from hatley castle and amazing hosts at the bed & breakfast to help accommodate our stay.
Is the description of this hotel not correct? Tell us


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Danny Wilson had many positives to speak of, but one overwhelming late negative as his side drew 3-3 with Gillingham on Tuesday night.
Danny Wilson Post-Gillingham
Gillingham 50/50 Results
Jay O'Shea Pre-Gillingham
Danny Wilson pre-Gillingham
On the Road: Bury
Right-back Paul McGinn joined a busy treatment room at the weekend after limping off with a hamstring injury during the second half away at Bury.
Away Tickets on Sale Now for Coventry and Southend
Liam Graham Post-Bury
Proact Car Boot Cancelled This Weekend
Bury: Extended Highlights
Danny Wilson Post-Bury
Danny Wilson aired his frustrations after his side failed to make the most of their opportunities in the 2-1 away loss to Bury on Saturday.
Shakers Versus Spireites Odds Preview
Bury Tickets on Sale at Gigg Lane
Spire Lotto Results, Week 22
YOUTH TEAM: Spireites Beat Worksop in Friendly
Kristian Dennis Pre-Bury
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Skip to comments.Danger Zone - (Haleigh Poutre)
Posted on 02/01/2006 7:29:10 AM PST by Ohioan from Florida
In the court (and courts) of life and death, a little 11-year-old Massachusetts girl named Haleigh Poutre could be the next Terri Schiavo. For those who have not heard the tragic story, Haleigh was beaten nearly to death last September, allegedly by her adoptive mother and stepfather. The beating left her unconscious and barely clinging to life.
Within a week or so of the beating, her doctors had written her off. They apparently told Haleigh's court-appointed guardian, Harry Spence, that she was "virtually brain dead." Even though he had never visited her, Spence quickly went to court seeking permission to remove her respirator and feeding tube. The court agreed, a decision affirmed recently by the supreme court of Massachusetts.
And so, no doubt with the best of intentions, a little girl who had already suffered so much was stripped by the Commonwealth of Massachusetts of even the chance to fight to stay alive. If she didn't stop breathing when the respirator was removed, which doctors expected, she would slowly dehydrate to death.
Close Call
Then came the unexpected:
(Excerpt) Read more at nationalreview.com ...
Ping. Does anyone know if someone has a Haleigh Poutre ping list?
There is a huge difference between this story and Schiavo. But you all know that.
very good article Ohioan!!
I didn't know where was a good place to end the excerpt. My apologies.
what's the difference? She is alive, brain damaged, and still some want to dehydrate her to death!
Of course there are differences, and there are similarities. But you all know that.
Then maybe I did excerpt it in the right place! I just want people to read the whole thing, and I would have posted the whole article, but I'm not up on all the posting article rules, and I don't want FR to get in trouble with copyright stuff.
The huge difference is that this young girl is still alive because the death ghouls have not succeeded in killing her, while Schiavo is forever dead.
I don't think anyone has a Heliegh-specific list, but I will ping my pro-life list.
ProLife Ping!
If anyone wants on or off my ProLife Ping List, please notify me here or by freepmail.
Can you add me to this list..
Isa 5:20: Woe unto them that call evil good, and good evil;...
I think it's sad that the media isn't paying attention to this either, but I hope they will as time goes by. Haleigh is far from out of danger.
Please don't think that Terri was 'brain dead' as was commonly reported by the media. She wasn't. She wasn't in a coma either. They misrepresented a lot about Terri.
With brain death, the person cannot even breathe on their own without the help of a respirator or ventilator. Terri did so for 15 years. She was no longer in a coma, as was commonly misreported early on. She came out from her coma within several weeks of her trauma, just as Randal McCloy, the surviving Sago miner, has recently done.
One, but not the only, difficulty for Terri was that of the Persistent Vegetative State. Whether Terri was truly in a PVS or had moved to another Minimally Conscious State was a point of debate even at her death. Smith brings up valid points in his article why it was only one thing that was problematic.
I hope that with more attention that Haleigh might have a brighter future.
There is a huge difference between this story and Schiavo. But you all know that
I am really glad you brought up this point. We can share it with all.
You will be glad to know there is an article stating just that! You can find it in the Boston Globe.
Haleigh Poutre is no Terri Schiavo
Moreover her brother made a fine response. Here it is.
Dear Ms. Young,
I writing in response to your column, Haleigh Poutre is no Terri Schiavo.
After reading all of the dishonesty that you purport as truths in regard to my sister Terri, I have to say that this is not only the most irresponsible article that I have yet to read from the media regarding Terris situation, but possibly the most egregious.
You wrote that, Schiavo had been in a persistent vegetative state for 15 years, and had undergone a barrage of tests showing that she had no higher brain functioning and no consciousness a fact on which all unbiased medical experts agreed.
I want to know how youve researched Terris case, and how youve obtained this information. Could it be that youve gathered your facts from the other opinion pieces similar to the one youve written about Terri?
If you are referring to autopsy that ostensibly proved that Terri was PVS, then you are entirely wrong. The Independent Medical Examiner who performed the autopsy, Dr. Jon Thogmartin, went to great lengths to caution those reviewing Terris autopsy that a post-mortem, forensic examination could neither confirm nor refute a diagnosis of persistent vegetative state. This is because PVS is a clinical diagnosis based on the behaviors of a living patient and not forensic pathology.
Moreover, Dr. Stephen Nelson a consulting pathologist who assisted Dr. Thogmartin with the autopsy report admitted that, while Terris symptoms were consistent with PVS, he could not rule out the possibility that Terri was in a minimally conscious state.
And there are numerous pathologists and physicians that have responded to the autopsy finding as being misleading and ambiguous. This information could have been and still can be easily provided to you.
I wonder if you were you aware that as indicated in Terris medical records she was talking when given rehabilitation in the months following her injury? This completely refutes the PVS diagnosis.
Additionally, did you know that there were more doctors on record with the court (from some of the most distinguished institutions from across the country) stating that Terri was not in PVS than there were doctors that said she was? Furthermore, most of these affidavits (there are more than 42 in total) submitted by these doctors said that if Terri was given proper therapy, her functioning capacity could have improved. Why was this never reported by the media?
Did you know that a recent British Medical Journal has reported that nearly 43% of all PVS diagnoses are, in fact, misdiagnoses? Based on this report, the Independent Medical Examiner findings, the sworn testimony and affidavits of over 42 physicians, neurologists and other healthcare professionals contradicting that Terri was in PVS, I believe it is incredibly reckless for you to make the sweeping statement that Terri was PVS.
The fact that my sister received no therapy for more than thirteen years, which also was rarely if ever reported by the media, only added to the deterioration of her condition. No autopsy report could have ever determined how much Terri would have improved if she was receiving proper rehabilitation and therapy during the past thirteen years.
To make the statement that, . . . all unbiased medical experts agreed as to the condition of my sister is so completely disingenuous that the only conclusion I can draw is that you did absolutely no research into the doctors that evaluated Terri.
Dr. Ronald Cranford, hired by Michael Schiavo, has testified in almost every major case that the removal of the feeding tube should be permitted. He even takes pleasure in the moniker, Dr. Humane Death. To call him unbiased is like calling Rush Limbaugh a liberal democrat. And one of the other doctors that testified in my sister trial, Dr. Peter Bambakidis, was later discovered as having ties to same organization as Michael Schiavos attorney George Felos.
But aside from all this, what is really sad is that your article and so many others like yours does nothing but discriminate against the disabled by labeling them with this dangerous and subjective PVS diagnosis. The PVS diagnosis was created for one reason and to serve only one purpose to kill those that are the weakest and most vulnerable among us.
No amount of damage to the brain or level of disability should give anyone the authority to kill. But this is exactly what you are saying - and is now happening all the time - that we can justify the killing of human beings because they have reached a level of profound brain damage that is unacceptable for us to care for them anymore. So, tell me, how long do you give Haliegh to improve before you give up on her and decide that she must die?
What is really shameful though is that I have to respond to articles like yours and defend my sisters life and explain why her family should have been permitted to love her.
Columnist Nat Hentoff commented on the media and how they have handled my sisters case. He said that it was so shoddy and inaccurate that it was the worst case of journalistic malpractice that hes seen in 25 years.
It continues.
Bobby Schindler
Thanks. :-)
God bless this little kid!
Never had anyone to love her, cherish her, etc....just people who want to pull the plug as fast as possible. God help us all.
PVS is a shamefully hopeless diagnosis. It is one where the doctor decides there is nothing he can do, and so gives himself permission to give up trying.
I am more encouraged by the doctors who understand that there are stages of consciousness, and that we can be unconscious for a while, then emerge from it, and sometimes retreat back into it, only to (oftentimes) re-emerge.
I detest when they refer to PVS as a permanent condition. Only death is permanent.
|...Lest anyone think that Haleigh's apparent consciousness protects her from suffering the fate of Terri Schiavo, who was ordered dehydrated by a Florida court based, in part, on a finding that she was unconscious, think again. In most states, exhibiting consciousness is not a defense against dehydration for profoundly impaired patients. Indeed, cognitively disabled people who are conscious are commonly dehydrated throughout the country. So long as no family member objects, the practice is deemed medically routine...
Sure, you're added to the Pro-Life ping list.
But those with an agenda are never interested in facts.
Thoughtful addition! :-)
I know you're a fan of Smith's, so I thought I should ping you to this thread. Hope you're doing well.
The futile care system is alive and well in our country as it stands today.
The Jihadists can find proof anywhere they want to look. Too bad they don't see their own faults and shortcomings, and are willing to kill others because of their blindness. I guess the same could be said of the euthanasiasts.
Imagine if we sentenced Scott Peterson to such a treatment.
The article in this thread describes the punishment of dehydration.
Defining dehydratable people.
It wasn't always so. It used to be thought of as unthinkable to remove a feeding tube. Then, as bioethicists and others among the medical intelligentsia began to worry about the cost of caring for dependent people and the growing number of our elderly and as personal autonomy increasingly became a driving force in medical ethics some looked for a way to shorten the lives of the most marginal people without violating the law or radically distorting traditional medical values. Removing tubes providing food and fluids was seen as the answer. After all, it was argued, use of a feeding tube requires a relatively minor medical procedure. Moreover, the nutrition provided the patient is not steak and potatoes, but a liquid formula prepared under medical auspices so as to ease digestion. There can also be complications such as diarrhea and infection.
Having reached consensus on the matter, the bioethics movement mounted a deliberate and energetic campaign during the 1980s to change the classification of ANH from humane care, which can't be withdrawn, to medical treatment, which can. The first people targeted for potential dehydration were the persistently unconscious or elderly with pronounced morbidity. Thus, bioethics pioneer Daniel Callahan wrote in the October 1983. Hastings Center Report, "Given the increasingly large pool of superannuated, chronically ill, physically marginalized elderly it [a denial of ANH] could well become the non treatment of choice."
And here is just one description of the effects of dehydration:
Saliva becomes thick and foul-tasting; the tongue clings irritatingly to the teeth and the roof of the mouth .... A lump seems to form in the throat ... severe pain is felt in the head and neck. The face feels full due to the shrinking of the skin. Hearing is affected, and many people begin to hallucinate... [then come] the agonies of a mouth that has ceased to generate saliva. The tongue hardens into what McGee describes as "a senseless weight, swinging on the still-soft root and striking foreignly against the teeth." Speech becomes impossible, although sufferers have been known to moan and bellow.
Next is the "blood sweats" phase, involving "a progressive mummification of the initially living body." The tongue swells to such proportions that it squeezes past the jaws. The eyelids crack and the eyeballs begin to weep tears of blood. The throat is so swollen that breathing becomes difficult, creating an incongruous yet terrifying
sense of drowning.
Finally ... there is living death, the state into which Pablo Valencia had entered when McGee discovered him on a desert trail, crawling on his hands and knees: "His lips had disappeared as if amputated, leaving low edges of blackened tissue; his teeth and gums projected like those of a skinned animal, but the flesh was black and dry as a hank of jerky; his nose was withered and shrunken to half its length, and the nostril-lining showing black; his eyes were set in a winkless stare, with surrounding skin so contracted as to expose the conjunctiva, itself as black as the gums...; his skin [had] generally turned a ghastly purplish yet ashen gray, with great livid blotches and streaks; his lower legs and feet ... were torn and scratched by contact with thorns and sharp rocks, yet even the freshest cuts were so many scratches in dry leather, without trace of blood" (Philbrick, 126-128).
Haleigh Poutre information
Please FReepmail me if you would like to be added to, or removed from, the Pro-Life/Pro-Baby ping list...
Of course there's a huge difference, but that doesn't matter to the willfully ignorant or the kookily obsessed.
Ted wanted water. He wanted to live.
Parents should not vacation in Florida with their adult children unless they trust them.
seriously. the medical providers are not healing, they are trafficking human beings straight to hospice.
Amerika, wake up. Ted was only 73 and people recover from strokes every single day!
Please put me on your ping list? Thanks.
Thanks for doing that. :-)
I'm calling Doctor Congressman Dave Weldon, Fl (federal or state investigation would be appropriate. Ted was from out of state but was murdered in Florida by Florida's DEATH CARE SYSTEM for the son.
It is a sad day for Ted Sr., a sad day for Florida, and for us all. Sadder still for all those condemned to horrible painful death by bioethicists, the ones we never hear about. Maybe we will be able to shine the spotlight on these murders.
Justice v. euthanasia in keywords. Okay to cover everybody subject to euthanasia here for the month of February? TERRI DAILIES for Jan can be pinged here.
We can become in effect the Haleigh ping list.
Here is the new Terri February Dailies thread.
Do you not see the similarites in the way the medical profession deems some as hopeless, or too expensive to care for?
Haleigh is lucky that she hasn't been denied food and water yet. Thank God that she was able to breathe without that respirator when push came to shove. The permission has already been granted to remove her feeding tube. It may still happen. Hopefully not.
If saying this makes me willfully ignorant or kookily obsessed, then I guess I'm guilty as charged. I'd rather be labelled either one, than to deny that there are similarities between Terri and Haleigh.
I noted the words "medical intelligentsia" with some irony.
I'm still here and don't plan to slow down!!
"They could still recommend to guardian Harry Spence that he withdraw her food and fluids lest she grow up profoundly disabled."
A court appointed guardian advocating death for his charge without ever having seen them, hospital staff brazenly lying to the state to get their patient killed; it's almost too much to comprehend sometimes. We are in a sad age.
In other words, imagine the Mother of all Hangovers. And guess what -- it's going to get worse for the rest of your life.
Disclaimer: Opinions posted on Free Republic are those of the individual posters and do not necessarily represent the opinion of Free Republic or its management. All materials posted herein are protected by copyright law and the exemption for fair use of copyrighted works.


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