The usefulness of ecotoxicological tools to improve the assessment of water bodies in a climate change reality.

This study aimed to analyse the added value of using ecotoxicological tools to complement and improve the assessment of natural water bodies status, in situations of climate change, with a higher frequency of extreme events as floods or droughts. Four water bodies of streams in the Guadiana Basin (Álamos, Amieira, Lucefécit, Zebro) were studied in 2017 and 2018 and classified based on the Water Framework Directive (WFD) parameters: Biological Quality Element - Phytobenthos (diatoms), General chemical and physicochemical elements, Specific pollutants, and Priority Substances. Complementarily, bioassays (including lethal and sublethal parameters) were carried out with organisms of different trophic levels: (i) the bacteria Aliivibrio fischeri; (ii) the microalgae Pseudokirchneriella subcapitata; (iii) the crustaceans Daphnia magna, Thamnocephalus platyurus and Heterocypris incongruens. A classification system with 5 scores was developed, permitting to classify water bodies from non-toxic (EC50 > 100 %; growth and feeding rate > 80 %; blue) to highly toxic (EC50 < 10 %; growth and feeding rate < 10 %; red). The comparison between the classification based on the WFD parameters and on ecotoxicological endpoints showed similar results for 71 % of the samples, and significant positive Pearson correlations were detected between the diatom-based Specific Polluosensitivity Index (SPI) and EC50V.fisheri, the algae growth rate and Shannon diversity index. These results indicate that when the biological quality elements cannot be used (namely under drought or flooding conditions) the application of ecotoxicological bioassays may be a good alternative. Further, when ecotoxicological parameters were included, an increase of worse classifications (Bad and Poor) was observed, revealing an improvement in the sensitivity of the classification, mainly in presence of specific and priority substances. So, the ecotoxicological analysis appears to provide useful information regarding the potential presence of both known and unknown contaminants at concentrations that cause biological effects (even within the WFD limits), in agreement with several authors that have already suggested its use in biomonitoring.

Liposomes co-encapsulating doxorubicin and glucoevatromonoside derivative induce synergic cytotoxic response against breast cancer cell lines.

Cancer is one of the main causes of death in the world and thus a global public health problem. Among the treatments available for cancer are surgery, radiotherapy, and chemotherapy. Currently, there is increased interest in the combination of two or more antitumor agents to achieve a synergistic effect in cancer therapy. Doxorubicin (DOX), a chemotherapeutic which has a potent antineoplastic action, has been used in the treatment of various tumors. However, the use of DOX is limited, mainly due to the cardiotoxicity. Therefore, nanostructured systems, such as liposomes, have been developed to carry this drug and target the tumor region, since tumor tissues present enhanced permeability and retention for nanosystems. Cardiac glycosides, such as digitoxin, have recently shown great antitumor potential despite the low therapeutic index which may limit their use. Furthermore, some compounds of this class have low water solubility, which makes their in vivo administration difficult. In this context, liposomes represent a valid strategy to carry simultaneously antitumor drugs allowing their intravenous administration. In this study, liposomes loaded with glucoevatromonoside containing peracetylated glucose hydroxyl groups (GEVPG) and DOX at molar ratio of 1:1 (SpHL-GEVPG:DOX 1:1) were developed, and their chemical and physicochemical properties were evaluated. This formulation presented a combination index (CI) lower than 1 at inhibitory concentration of 90 % growth (IC90) for three human breast tumor lines evaluated (0.52 ± 0.39 for MDA-MB-231, 0.19 ± 0.13 for MCF-7, and 0.99 ± 0.09 for SKBR-3). These results indicate a synergistic cytotoxic effect of the GEVPG and DOX combination encapsulated in liposomes. In addition, SpHL-GEVPG:DOX 1:1 presented selectivity towards these cancer cells. Long-term in vitro cytotoxicity studies demonstrated that MDA-MB-231 surviving cells after treatment with SpHL-GEVPG:DOX 1:1 did not recover proliferation capacity after 21 d. From the studies of cell cycle and death pathway evaluation, it was observed that SpHL-GEVPG:DOX 1:1 arrested the cell cycle in the G2/M phase and similarly induced apoptosis and necrosis. However, SpHL-GEVPG:DOX at molar ratio of 1:1 showed lower induction of both apoptotic and necrotic pathways compared to free DOX and SpHL-DOX, suggesting that the mechanism of death involved may not be related to necrosis or apoptosis. Lastly, SpHL-GEVPG:DOX 1:1 showed a good storage stability for 90 d at 4 °C. Therefore, the results of the present work indicate the potential use of SpHL-GEVPG:DOX 1:1 as a new anticancer formulation.

Long-circulating and fusogenic liposomes loaded with a glucoevatromonoside derivative induce potent antitumor response.

Cancer is an important public health problem, being one of the leading causes of death worldwide. Most antineoplastic agents cause severe toxic effects and some types of cancer do not respond or are resistant to the existing pharmacotherapy, necessitating the research and development of new therapeutic strategies. Cardenolides have shown significant antitumor activity due to their ability to inhibit the Na+K+ATPase enzyme, and the expression of this enzyme is increased in tumor cells. Glucoevatromonoside containing peracetylated glucose hydroxyl groups (GEVPG) is a cardenolide derivative that has low solubility in aqueous media, which constitutes a barrier to its potential biological applications. In this context, the use of liposomes represents a promising strategy to deliver GEVPG, thus allowing its intravenous administration. In this study, long-circulating and fusogenic liposomes containing GEVPG (SpHL-GEVPG) were developed, and their chemical and physicochemical properties were evaluated. SpHL-GEVPG presented adequate properties, including a mean diameter of 182.2 ± 2.7 nm, a polydispersity index equal to 0.36 ± 0.03, a zeta potential of -2.37 ± 0.31 mV, and a GEVPG entrapment of 0.38 ± 0.04 mg/mL. Moreover, this formulation showed a good stability after having been stored for 30 days at 4 °C. The cytotoxic studies against breast (MDA-MB-231, MCF-7, and SKBR-3) and lung (A549) cancer cell lines demonstrated that SpHL-GEVPG treatment significantly reduced the cell viability. In addition, the SpHL-GEVPG formulation presented a good selectivity toward these cancer cells. The evaluation of the therapeutic efficacy of the treatment with SpHL-GEVPG showed a potent anticancer effect in an A549 human lung cancer xenograft model. SpHL-GEVPG administered at doses of 1.0 and 2.0 mg/kg (i.v.) induced antitumor effect comparable to paclitaxel given at dose of 10 mg/kg (i.v.) to mice. Therefore, the results of the present work indicate the potential applicability of SpHL-GEVPG as a new anticancer formulation.

Characterization of the P373L E-cadherin germline missense mutation and implication for clinical management.

AIM: Hereditary diffuse gastric cancer (HDGC) is a cancer susceptibility syndrome caused by E-cadherin germline mutations. One-third of these mutations are of the missense type, representing a burden in genetic counselling. A new germline missense mutation (P373L) was recently identified in a HDGC Italian family. The present work aimed at addressing the disease-causative nature of the P373L mutant. METHODS: Assessment of the P373L mutation effect was based on cell aggregation and invasion assays. LOH analysis at the E-cadherin locus, search for somatic E-cadherin mutations and for promoter hypermethylation were performed to identify the mechanism of inactivation of the E-cadherin wild-type allele in the tumour. RESULTS: In vitro the P373L germline mutation impaired the E-cadherin functions. E-cadherin promoter hypermethylation was observed in the tumour of the P373L mutation carrier. CONCLUSION: We conclude that the combination of clinical, in vitro and molecular genetic data is helpful for establishing an accurate analysis of HDGC-associated CDH1 germline missense mutations and subsequently for appropriate clinical management of asymptomatic mutation carriers.

Studies on pharmaceutical ethnobotany in Arrabida Natural Park (Portugal).

An ethnobotanical survey was carried out in Arrabida Natural Park, a Portuguese Protected Area in the Southwest of the Iberian Peninsula, with an area of 10,820 ha. Working with 72 local people, data on medicinal uses of 156 taxa, belonging to 56 botanical families, were obtained and presented, of which 214 uses corresponding to 81 taxa were previously unreported.

An international perspective on the treatment of pediatric shock: the Brazilian experience.

Shock in the pediatric population has many preventable causes. Treatment of children in shock will depend on access to health services, training of health personnel, availability of diagnostic procedures, monitoring, and therapeutic measures. Countries will differ among themselves and within themselves in the care provided to children developing shock. In Brazil, the majority of children are cared for in public hospitals, which often lack resources for basic care. Many children in shock do not even reach healthcare services. Investment in training healthcare personnel in a simplified and systematic approach to shock and access to equipped health services are basic to improved outcomes in the treatment of pediatric shock. The Brazilian experience in the treatment of children in shock outside hospital facilities, in the emergency department, and in the ICU is described.

Column preconcentration of organotin with tropolone-immobilized and their determination by electrothermal atomization absorption spectrometry.

A method for the determination of total organic tin from marine water samples by electrothermal atomization absorption spectrometry (ETAAS) is described. Samples are previously preconcentrated with a chelating molecule (tropolone) impregnated on a macroporous polymer (Amberlite XAD-2). The graphite furnace programme and preconcentration parameters were optimized. Calibration and addition graphs were performed. Sensitivity obtained with this procedure was 13 ng l(-1). Relative standard deviation was always >10% and analytical recovery were satisfactory, approximately 100%. Some possible interferences were investigated, having no problems with this factor. This procedure allows the distinction between organotin compounds and inorganic tin IV, since the latter is not retained on the column.

Visual function in neurofibromatosis.

OBJECTIVES: To evaluate the visual function in patients with neurofibromatosis (NF) and to study the etiology and incidence of visual dysfunction associated with NF. PATIENTS AND METHODS: A total of 75 patient with diagnostic criteria for NF were evaluated. Neuro-ophthalmological examination as well as electrophysiological and imaging studies were performed. Special attention was given to the presence of visual dysfunction and to its correlation with the ophthalmic changes that were found. RESULTS: Ocular findings were present in 42 (56%) patients. Visual dysfunction was identified in only 11 (14.7%) patients. Visual acuity decrease was the most prevalent change, being present in eight (72.7%) of patients with visual dysfunction. Nystagmus, strabismus, visual field defects, and color vision defects were also detected. Therapy is also reviewed. The prognosis of the 11 patients with visual dysfunction was unfavorable, and is discussed. CONCLUSION: The prevalence of NF and ophthalmological findings related to it make the problem of visual dysfunction in NF a serious one, deserving of the attention of all ophthalmologists. Clinical examination, associated with complementary diagnostic techniques (mainly imaging studies), allows NF's identification, definition, and therapy. The high prevalence of asymptomatic ocular findings in NF (73.8%) highlights the role of imaging techniques in its evaluation.