BACKGROUND:Facial disfigurement has been considered one of the most challenging consequences of the surgical treatment for head and neck cancer patients, mainly due to the importance of the facial region for the personal identity, body self-image and interpersonal interactions, which might affect negatively the quality of life. The aim of this study was to assess factors associated with the quality of life of subjects with facial disfigurement due to surgical treatment.MATERIAL AND METHODS:Clinical data were retrieved from 103 patient's medical records and quality of life data were collected using the Functional Assessment of Cancer Therapy (FACT-H&N) questionnaire. Moreover, the degree of facial disfigurement was classified by means of a specific ordinal scale.RESULTS:Data from the FACT-H&N questionnaire showed that the domain directly related to head and neck symptoms was considered the most impacted, while emotional domain was the least affected. Lower quality of life was associated with sequels in the neck and/or lower third of the face (β=-0.39; p=0.001), a higher level of disfigurement (β=-0.29; p=0.016) and female gender (β=-0.20; p=0.038).CONCLUSIONS:Disfigurement due to surgical treatment was significantly associated with the functional dimension of the patients, especially in extensive sequels in the cervical and lower regions of the face.

The mode of action for nanoparticle (NP) toxicity in aquatic organisms is not yet fully understood. In this work, a strategy other than toxicity testing was applied to Daphnia magna exposed to TiO2-NPs: the use of nuclear microscopy and the assessment of protein profile. D. magna is a keystone species broadly used as a model system in ecotoxicology. Titanium (Ti) was found in the D. magna digestive tract, mainly in the gut. The penetration of Ti into the epithelial region was greater at higher exposure levels and also observed in eggs in the brood pouch. The protein profile of individuals exposed to different concentrations showed that 2.8 and 5.6 mg/L TiO2-NP concentrations induced an over-expression of the majority of proteins, in particular proteins with molecular weight of ∼120, 85 and 15 kDa, while 11.2 mg/L TiO2-NP had an inhibitory effect on protein expression. The Matrix-assisted laser desorption ionization with tandem time of flight mass spectrometry (MALDI-TOF/TOF MS) analysis of these proteins consistently identified them as vitellogenin (Vtg)-like proteins, associated with enzymes involved in redox balance. These results indicate that Vtg-like proteins are up-regulated in D. magna exposed to TiO2-NPs. Vitellogenesis is associated with the reproduction system, suggesting that TiO2-NP exposure can impair reproduction by affecting this process. The precise mode of action of TiO2-NPs is still unclear and the results from this study are a first attempt to identify specific proteins as potential markers of TiO2-NP toxicity in D. magna, providing useful information for future research.

The new properties of engineered nanoparticles drive the need for new knowledge on the safety, fate, behavior and biologic effects of these particles on organisms and ecosystems. Titanium dioxide nanoparticles have been used extensively for a wide range of applications, e.g, self-cleaning surface coatings, solar cells, water treatment agents, topical sunscreens. Within this scenario increased environmental exposure can be expected but data on the ecotoxicological evaluation of nanoparticles are still scarce. The main purpose of this work was the evaluation of effects of TiO2 nanoparticles in several organisms, covering different trophic levels, using a battery of aquatic assays. Using fish as a vertebrate model organism tissue histological and ultrastructural observations and the stress enzyme activity were also studied. TiO2 nanoparticles (Aeroxide® P25), two phase composition of anatase (65%) and rutile (35%) with an average particle size value of 27.6±11 nm were used. Results on the EC50 for the tested aquatic organisms showed toxicity for the bacteria, the algae and the crustacean, being the algae the most sensitive tested organism. The aquatic plant Lemna minor showed no effect on growth. The fish Carassius auratus showed no effect on a 21 day survival test, though at a biochemical level the cytosolic Glutathione-S-Transferase total activity, in intestines, showed a general significant decrease (p<0.05) after 14 days of exposure for all tested concentrations. The presence of TiO2 nanoparticles aggregates were observed in the intestine lumen but their internalization by intestine cells could not be confirmed.

Resumo Objectivo: Avaliar a incidência e os factores de risco de lesão renal após um primeiro episódio de pielonefrite aguda (PNA). Métodos: Foram analisados os processos clínicos de crianças seguidas na Consulta de Nefrologia Pediátrica de um Hospital de Nível III, após internamento por PNA entre Janeiro de 2000 e Dezembro de 2002. Foram excluídas as crianças com antecedentes de infecção urinária, uropatía previamente conhecida e aquelas que não completaram a investigação complementar imagiológica e funcional. Resultados; 86 crianças cumpriam os critérios de inclusão, 57% do sexo feminino e 59% com idade igual ou inferior a 1 ano. A Esckeríchia coli foi o agente isolado cm 92% dos casos. Em 21 casos (24%) foi encontrada lesão renal sequelar. Num total de 22 crianças (25%) que apresentavam refluxo vésico-urcteral (RVU), 41% linham lesão renal. 12 crianças (19%), num total de 64 crianças sem RVU, tinham lesão renal. A presença de RVU correlacionou-se com a existência de cicatriz de uma forma estatisticamente significativa (p 0.037). Verificámos uma taxa de abandono da Consulta de 37%. Conclusão: A lesão renal, após uma primeira PNA, acontece em cerca de um quarto das crianças, sendo o RVU o factor de risco mais importante. Palavras-Chave: pielonefrite aguda, cicatriz renal, refluxo vésico-urcteral.

The toxicity of titanium dioxide nanoparticles (TIO2 NPs) and oxidative stress effects were studied in two freshwater fish species (Carassius auratus and Danio rerio) exposed for 21 days to different concentrations (0.01, 0.1, 1, 10, 100/mgL) of TIO2 NPs and to a control (tap water). Additional fish were transferred to clean water for 14 days to assess the ability to recover from exposure to TIO2 NPs. Activities of the enzyme glutathione-S-transferase (GST) and lipid peroxidation (LPO) (malondialdheyde) were measured as indicators of oxidative stress. Histological and ultra-structural changes in livers from bothspecies of fish were evaluated by light and electron microscopy. Results show a general GST activity increase according to TIO2 NPs concentrations, which is in agreement with data from LPO. After 21 days, GST activities decreased possibly caused by suppression of GST synthesis as a result of severe stress. Histological and ultra-structural analysis of livers from exposed fish show degeneration of the hepatic tissue and alterations in hepatocytes such as glycogen depletion and an increase in lipofucsin lysosome-like granules. After a depuration period a partial recovery for biochemical markers and cells was observed. The results suggest that TIO2 promotes alterations in hepatic tissues compatible with oxidative stress.

Engineered nanomaterials such as nanoparticles (NPs) are increasingly being used for commercial purposes in products within medicine, electronics, sporting goods, tires, textiles and cosmetics. They comprise diverse types of materials from metals, polymers, ceramic to biomaterials and have been defined as particles with at least one dimension in the order up to 100 nm1. The higher toxicological potential of NPs is mostly due to their small size, wide surface, increase of their chemical reactivity and biological activity and the capacity to generate free radicals. NPs also can have the ability to penetrate trough the biological barriers and to move easily through the biological systems. Therefore, is mandatory to assess the toxicity of these nanomaterials, since their industrial production and uses will also result in releases to the environment with unclear consequences.The aim of the present work is to evaluate the toxicity of titanium dioxide (TiO2) NPs on gill gluthatione-S-transferase activity (GST), lipid peroxidation and on structure of the gills of two freshwater fish species (Carassius auratus and Danio rerio). Suspensions of TiO2 NPs, with an average size of 21 nm, were prepared using distillate water and then ultrasonicated (10 min, 35 KHz). The suspensions were added to 10L of tap water in exposure tanks, to obtain nominal concentrations (0.01; 0.1; 1, 10; 100 TiO2 mg/L). The test fish, C. auratus (N=144) and D. rerio (N=80), were randomly distributed by 6 exposure tanks and an additional tank with clean tap water was used as control. Fish were sampled after 7, 14, and 21 days. Six fish from both species were left for depuration in clean tap water during 14 days and then sacrificed. The GST activity was determined by following the procedure described by Habig et al. and lipid peroxidation was measured based on the Thiobarbituric Acid Reactive Species method. The tissues were processed essentially according to Martoja and Martoja for light microscopy (LM). For transmission electron microscopy (TEM) the samples were fixed sequentially in glutaraldehyde, osmium tetroxide and uranyl acetate, dehydrated in ethanol and embedded in Epon-Araldite according to standard procedures. The histological and ultrastructural observations were performed using a Leica-ATC 2000 microscope and a JEOL 100-SX electron microscope respectively.The results show a significant increase of GST in gills tissues for C.auratus exposed to 10 and 100 mg/L TiO2 NPs and a decrease following the depuration period. With respect to D. rerio a significant increase was observed in fish exposed to 1, 10 and 100 mg/L TiO2 NPs. Lipid peroxidation are in agreement with GST results but showing a significant increase for fish (C.auratus and D. rerio) exposed to concentrations of 0.1 TiO2 mg/L NPs and higher. Usually, the oxidative stress caused by exposure to TiO2 NPs is attributed to hydroxyl radicals (OH) generated by photochemical (UV/vis) processes but it may be also related to specific properties of TiO2 NPs such as size, surface area and solubility that can influence the degree of toxicity. The results from LM observations (Fig. 1) showed that exposure to TiO2 NPs affected gill tissues, with changes being detected in both fish species exposed to 0.1 TiO2 mg/L NPs and higher which is in accordance with biochemical results. Changes include different degrees of hyperplasia (from low to complete fusion of lamellae). The TEM analysis revealed that TiO2 NPs were internalized by gills epithelial cells accumulating in vacuoles inside these cells (Fig. 2). After the depuration period it was observed that the capability for gills to recover was not complete. The results show a strong response to oxidative stress caused by exposure to TiO2 NPs, possibly because they are in direct contact with the exposure medium and function as a first barrier against external aggression. However, the gills changes observed following exposure and a partial recover after depuration suggest that TiO2 NPs may cause deleterious effects in fish gills compromising fish homeostasis.The authors acknowledge the funding by Fundação para a Ciência e Tecnologia through grant PTDC/CTM/099446/2008 and through project no. PEst-C/EQB/LA0006/2011 granted to Requimte.

Nanoparticles (NPs), particles with at least one dimension less than 100 nm, are used in many industrial applications and to produce new types of materials with unique physicochemical properties. The aquatic environment is commonly the ultimate recipient for NPs and there is uncertainty of exposure as understanding and data regarding the potential detrimental effects of NPs on aquatic biota are missing. In this study, titanium dioxide (TiO2) was chosen for its potential use in technology and diverse industrial applications. The objective of this work is to evaluate the toxicity of TiO2 NPs on total liver glutathione-S-transferase (GST), lipid peroxidation and tissue structure of the livers of two freshwater fish species (Carassius auratus and Danio rerio). Stock suspensions of TiO2 NPs, with an average size of 21 nm, were prepared using distilled water and then ultrasonicated (10 min, 35 KHz). The suspensions were added to 10L of tap water in exposure tanks, to obtain nominal concentrations (0.01; 0.1; 1, 10; 100 TiO2 mg/L). The test fish, C. auratus (N=144) and D. rerio (N=80), were randomly distributed by 6 exposure tanks and an additional tank with clean tap water was used as control. Fish were sampled after 7, 14, and 21 days. Six fish from both species were left for depuration in clean tap water during 14 days and then sacrificed. Immediately after sampling the fish were processed for enzymatic determination and histopathology. The GST activity was determined by following the procedure described by Habig et al. and lipid peroxidation was measured based on the Thiobarbituric Acid Reactive Species method. The tissues were processed essentially according to Martoja and Martoja for light microscopy (LM). For transmission electron microscopy (TEM) the samples were fixed sequentially in glutaraldehyde, osmium tetroxide and uranyl acetate, dehydrated in ethanol and embedded in Epon-Araldite according to standard procedures. The histological and ultra-structural observations were carried out using a Leica microscope (Leica-ATC 2000) and a JEOL 100-SX electron microscope respectively. The results showed increased activities of the GST in livers with increasing TiO2 NP concentrations after 7 days of exposure, however after 14 days a trend to decrease was observed for both species. The GST results suggest that the increase of activity of these detoxification enzymes can be a response to oxidative stress caused by the generation of reactive oxygen species by the NP. On the other hand, size, chemical composition, surface area, shape, solubility and aggregation may also contribute for NPs toxicity. The results from lipid peroxidation showed an increase according to tested concentrations suggesting that TiO2 NPs is able to cause cell damage and is in agreement with biochemical and histological findings. After 14 days of depuration, GST and lipid peroxidation levels were not significant different from controls suggesting that cells are able to recover in a certain degree. The results from LM (Fig. 1) showed that exposure to TiO2 NPs affected liver structure, with more pronounced changes detected in livers from fish exposed to higher concentrations. Observed changes include tissue degeneration, inflammation and pyknosis among others. The TEM analysis revealed also severe changes in liver cells compatible with oxidative stress. Hepatocytes of treated fish showed glycogen depletion, swollen mitochondria and increased lysosomes, compared to controls. After depuration, some cells recovered nearly normal morphology, but retained the lysosomes, while others underwent necrotic changes (Fig. 2). Differences among the two species studied were of a quantitative nature, and more pronounced in Danio rerio. The results suggest that potential risk to fish health exist related to the TiO2 NPs release to the aquatic environment and may cause deleterious effects in aquatic organisms. It is evident that the effects of TiO2 NPs on environment is a matter of great concern and the precise mechanisms of toxicity of this and other types of NPs must be clarified. The authors acknowledge the funding by Fundação para a Ciência e Tecnologia through grant PTDC/CTM/099446/2008 and through project no. PEst-C/EQB/LA0006/2011 granted to Requimte.

Wastewater management has a central role in sustainable development, and, in this context, an integrated management of wastewater treatment plants (WWTP) can be important. WWTP discharge complex effluents and for a new strategy in environmental protection ecotoxicological evaluation should complement the usual chemical evaluation. The EU project WW4Environment was set up for a WWTP located in Lisbon area and discharging into Tagus estuary (Portugal). One of the main objectives of the project is to optimize the management of the WWTP in terms of environmental impact. A battery of toxicity tests with organisms bearing different functions at the ecosystem level (the bacterium Vibrio fischeri, the alga Pseudokirchneriella subcapitata, the crustaceans Thamnocephalus platyurus and Daphnia magna, and the plant Lemna minor) was used to characterize the wastewater in the different treatment phases. V. fischeri, test organism for Microtox test, was the most sensitive species in WWTP samples evaluation. Microtox, Alga, and Daphnia tests were able to distinguish two levels of treatment and to assess toxicity removal efficiency. The results demonstrated not only that the treatment efficiently reduced wastewater toxicity, but also that the use of an ecotoxicological approach can contribute to the environmental management of the treatment plant.

Engineered Titanium nanoparticles are used for a wide range of applications from coatings, sunscreen cosmetic additives to solar cells or water treatment agents. Inevitably environmental exposure can be expected and data on the ecotoxicological evaluation of nanoparticles are still scarce.The potential effects of nanoparticles of titanium dioxide (TiO2) on two model organisms, the water flea, Daphnia magna and the duckweed Lemna minor, were examined in semichronic toxicity tests.Daphnia and Lemna were exposed to TiO2 nanoparticles (average particle size value of 28 +/- 11 nm (n = 42); concentration range, 1.4-25 mg/L) by dietary route and growth in medium containing the nanoparticles of TiO2, respectively. Both morphology and microdistribution of Ti in the individuals were examined by nuclear microscopy techniques. A significant amount of TiO2 was found accumulated in Daphnia exposed to nanoparticles. Nuclear microscopy imaging revealed that Ti was localized only in the digestive tract of the Daphnia, which displayed difficulty in eliminating the nanoparticles from their body. Daphnia showed higher mortality when exposed to higher concentrations of TiO2 (>10 mg/L). The exposure to TiO2 nanoparticles above 25 mg/L caused morphological alterations in Lemna. The roots became stiff and fronds colorless. The Ti mapping of cross-sections of roots and fronds showed that Ti was mainly deposited in the epidermis of the fronds and roots, with minor internalization.In summary, exposure of aquatic organisms to TiO2 nanoparticles may alter the physiology of these organisms at individual and population levels, posing risks to aquatic ecosystems. (C) 2013 Elsevier B.V. All rights reserved.

Pollution control has been changed by advances in scientific knowledge, because there is a connection of environmental contamination with the ability to measure it. With greater understanding of the impact of wastewater on the environment and more sophisticated analytical methods, advanced treatment is becoming more common (Lofrano & Brown, 2010). The assessment of biological effects of wastewater discharges in the ecosystems is today considered relevant and ecotoxicological tests identifying the ecological hazard are useful tools for the identification of environmental impacts. Direct toxicity assessment, making use of ecotoxicological tests, can play an important role in supporting decision-making, either regulatory driven or on a voluntary basis. Within the Integrated Pollution Prevention and Control Directive IPPC, 2008/1/EC (European Commission [EC], 2008), the Direct Toxicity Assessment concept has been included as a suitable monitoring tool on effluent in several Best Available Techniques (BAT) Reference Documents. Also, in Water Framework Directive – WFD, 2000/60/EC (EC, 2000), direct toxicity assessment of Wastewater Treatment Plant (WWTP) effluents can contribute to attain or keep ecological quality objectives in water masses. So, for EU countries to comply with good ecological status, ecotoxicity evaluation of WWTP effluents is extremely relevant. In many countries ecotoxicity tests are already in use for wastewater management (Power & Boumphrey, 2004; Tinsley et al., 2004; United States Environmental Protection Agency [USEPA], 2004; Vindimian et al., 1999). Bioassays are also used for wastewater surveillance and BAT compliance by authorities in Germany (Gartiser et al., 2010a). A global evaluation of wastewaters should include ecotoxicological tests to complement the chemical characterization, with advantages especially in the case of complex wastewaters (Mendonca et al., 2009). This approach has advantages particularly to protect biological treatment plants from toxic influents (Hongxia et al., 2004), to monitor the effectiveness of WWTP (Cēbere et al., 2009; Daniel et al., 2004; Emmanuel et al., 2005; Libralato et al., 2006; Metcalf & Eddy, 2003) and in the impact assessment of complex wastewaters. Bioassays are considered a suitable tool for assessing the ecotoxicological relevance of complex organic mixtures (Gartiser et al., 2010b).