Cell-free DNA analysis in current cancer clinical trials: a review.

Cell-free DNA (cfDNA) analysis represents a promising method for the diagnosis, treatment selection and clinical follow-up of cancer patients. Although its general methodological feasibility and usefulness has been demonstrated, several issues related to standardisation and technical validation must be addressed for its routine clinical application in cancer. In this regard, most cfDNA clinical applications are still limited to clinical trials, proving its value in several settings. In this paper, we review the current clinical trials involving cfDNA/ctDNA analysis and highlight those where it has been useful for patient stratification, treatment follow-up or development of novel approaches for early diagnosis. Our query included clinical trials, including the terms 'cfDNA', 'ctDNA', 'liquid biopsy' AND 'cancer OR neoplasm' in the FDA and EMA public databases. We identified 1370 clinical trials (FDA = 1129, EMA = 241) involving liquid-biopsy analysis in cancer. These clinical trials show promising results for the early detection of cancer and confirm cfDNA as a tool for real-time monitoring of acquired therapy resistance, accurate disease-progression surveillance and improvement of treatment, situations that result in a better quality of life and extended overall survival for cancer patients.

A NOVEL BIOLOGICAL DOSIMETRY ASSAY AS A POTENTIAL TOOL FOR TRIAGE DOSE ASSESSMENT IN CASE OF LARGE-SCALE RADIOLOGICAL EMERGENCY.

In case of mass radiological emergencies, new strategies involving biological and clinical endpoints are requested for an efficient triage classification of casualties. For this purpose, we developed a novel protocol combining the two most established cytogenetic methods used in biological dosimetry (dicentric and micronucleus assays) into a single one, in order to have a time-saving, inexpensive and potentially automatable instrument to be used for triage purposes in case of large-scale radiological events. This method could be considered as a 'three in one' assay allowing the simultaneous scoring of chromosome aberrations and micronuclei on a single slide, and also enabling to discriminate between metaphases in first and second cell division without the Fluorescence plus Giemsa staining. This method needs further validation through inter-comparisons involving biological dosimetry laboratories, to verify its reproducibility. Moreover, the possibility to apply the already existing software for automation for dicentric and micronucleus assays could be also verified.

Role of the adjunctive antimicrobial photodynamic therapy to periodontal treatment at plasmatic oxidative stress and vascular behavior.

BACKGROUND: To evaluate for the first time in vivo the effects of methylene blue (MB) photosensitizer dissolved in ethanol in antimicrobial photodynamic therapy (aPDT) as adjuvant periodontal treatment, at plasmatic oxidative stress and vascular behavior in rat model. METHODS: Wistar rats were divided into negative control (NC, no periodontitis) and positive control (PC, with periodontitis, without any treatment). The other groups had periodontitis and were treated with scaling and root planing (SRP); SRP+aPDT+MB dissolved in water (aPDT I); SRP+aPDT+MB dissolved in ethanol (aPDT II). The periodontitis was induced by ligature at the mandibular right first molar. At 7/15/30days, rats were euthanized, the plasma was used to determine oxidative stress parameters and gingival tissue for histomorphometric analysis. RESULTS: PC showed higher thiobarbituric acid reactive substances levels in 7/15/30days. aPDT II was able to block the lipid peroxidation, especially between 15th and 30th days. Glutathione reduced levels were consumed in PC, aPDT I and II groups throughout the experiment. aPDT II increased the vitamin C levels which were restored in this group in the 30th day. aPDT II group showed the highest number of blood vessels. CONCLUSION: In summary, the aPDT with MB dissolved in ethanol provides better therapeutic responses in periodontitis treatment.

Mujeres privadas de libertad y apego: una intervención de terapia ocupacional en Magallanes / Women prisoners and attachment: an occupational therapy intervention in Magallanes

Esta investigación se fundamente desde una perspectiva de derechos humanos y enfoque de género. Su objetivo fue explorar el impacto de una intervención de Terapia Ocupacional sobre el vínculo madre e hija en mujeres privadas de libertad del Centro Penitenciario de Punta Arenas, región de Magallanes. Para este estudio se empleó la metodología cualitativa y su diseño de investigación en un estudio de caso. Utilizó técnicas de observación participante y entrevistas en profundidad. El análisis de los datos se desarrolló bajo la directriz de la teoría fundamentada. Los resultados indican cambios positivos en las áreas de afectividad, motivación y empatía. Se evidencia un impacto positivo en el apego, desarrollo del vínculo madre-hija, toma de conciencia de la relación filial y cambios positivos en la percepción del rol de madre. Resulta urgente seguir avanzando a nivel nacional en el reconocimiento y abordaje de necesidades de las mujeres que residen en el sistema penitenciario, así como en estrategias que les permitan acceder a ocupaciones significativas en este contexto, respetando sus derechos humanos fundamentales.

This research is based in human rights perspectives with gender approach. The purpose was to evaluate the impact of an occupational therapy intervention regarding relationship between mother and child inside the Penitentiary Precinct of Punta Arenas. In order to accomplish the goal qualitative methods were used and the design was a case study. Data collection included active-participant observation and interviews with mothers. Data analysis was developed under the guidance of grounded theory. Results show positive changes in affection, attachment, motivation and empathy. There is a positive impact in attachment, development of the link mother-daughter, conscience of the affiliated relation and positive changes in the perception of the mother’s role. It is urgent to continue advancing in Chile about the recognition and boarding on needs of women who reside in the penitentiary system: in strategies that allow them to accede to significant occupations in this context, and therefore to respect their fundamental human rights.

Non-motor symptoms and cardiac innervation in SYNJ1-related parkinsonism.

INTRODUCTION: PARK20 is a rare autosomal recessive parkinsonism related to the SYNJ1 gene and characterized by early-onset of disease and atypical signs such as supranuclear vertical gaze palsy, dementia, dystonia, and generalized tonic-clonic seizures. OBJECTIVE: Non-motor features and cardiac sympathetic innervation were assessed in two siblings affected by parkinsonism who harboured the homozygous Arg258Gln mutation in the SYNJ1 gene. METHODS: The Non-Motor Symptoms, the SCOPA-AUT, the Mayo Sleep Questionnaires and polysomnography were used to investigate non-motor signs (NMS), autonomic dysfunction and REM Behavioural Disorder (RBD). Cognitive functions were examined by an extensive battery of neuropsychological tests. In addition, motor and sensory nerve conduction studies and evoked laser potentials were performed. Cardiac sympathetic innervation was assessed in the two patients by (123)I-metaiodobenzylguanidine (MIBG) scintigraphy, computing early and late heart-to-mediastinum (H/M) ratios and myocardial washout rates (WR). RESULTS: Among the non-motor symptoms and autonomic signs, case 1 had cold intolerance, drooling and dysphagia, while case 2 had pain and urinary dysfunction. Both cases showed mood and behavioural disorders. RBD were not found, whereas the neuropsychological assessment revealed a progressive cognitive impairment. Neurophysiological studies revealed no abnormalities. Indexes of cardiac sympathetic innervation in the two patients did not differ from those of control subjects. CONCLUSIONS: Our findings expand the phenotypic profile of SYNJ1-related parkinsonism. Preserved cardiac sympathetic function and absence of RBD suggest that PARK20 should be explained by a pathogenic mechanism different from Lewy Body pathology, or that the latter is not as widespread as idiopathic Parkinson's disease.

Hidatidosis cardiaca: caso clínico / Cardiac hydatidosis: Report of one case

Background: Hydatidosis is an endemic zoonosis in Chile. We report a 48-year-old former slaughterman, with a previous history of pulmonary hydatidosis, who presented a stroke without associated cardiovascular symptoms. An echocardiogram revealed a tumor mass with cystic features in the left ventricle. The patient was operated and the cyst was successfully excised. During the follow up, the patient remains asymptomatic.

Sonic hedgehog stimulates proliferation of human periodontal ligament stem cells.

Regulation of cell renewal in the periodontium is a critical cell function that has not been clarified. Sonic hedgehog (Shh) is a secreted signaling molecule that plays a key role during development and adult tissue homeostasis. In the present study, we have analyzed the role played by Shh in human periodontal ligament stem cell (HPLSC) proliferation. HPLSC were isolated with anti-STRO-1 antibodies. Shh increased the expression of GLI1 and PTC-1 and selectively stimulated cell proliferation in STRO-1(+) derived from adult periodontal ligament. Shh components were localized to primary cilia in STRO-1(+) cells after Shh stimulation. STRO-1(+) also expressed Shh, suggesting an autocrine-regulated phenomenon. Thus, we propose that Shh plays a critical role in the regulation of cell proliferation in STRO-1(+)/HPLSC.

Intensification of gas-phase photoxidative dehydrogenation of ethanol to acetaldehyde by using phosphors as light carriers.

In this work a significant improvement of VO(x)/TiO(2) photocatalytic activity in the selective partial oxidation of ethanol to acetaldehyde was achieved by the simultaneous irradiation with light emitting phosphorescent particles and UVA-LEDs as external light source. Photocatalytic tests were carried out in a gas-solid photocatalytic fluidized bed reactor at high illumination efficiency, in which the bed is constituted by VO(x)/TiO(2) photocatalyst at nominal V(2)O(5) content of 5 wt% and suitable selected phosphors, diluted with glass spheres. In this way, phosphors were fluidized together with the catalyst, excited by external UVA-LEDs, emitting their stored energy in close proximity to the catalyst. In the absence of phosphors the ethanol consumption rate initially grows linearly with initial alcohol concentration, then bends towards an asymptotic value for initial ethanol concentration higher than 0.5 vol%. By contrast, when phosphors are present, the ethanol consumption rate increased linearly in the overall range. In all cases acetaldehyde was the main product detected in gas phase with a selectivity of about 97%, ethylene and carbon dioxide the by-products. The results evidenced that the presence of phosphors allowed improved photon transfer, increasing the apparent quantum yield from 2 to 30% together with a high photoreactivity.

An atypical clinical presentation for the first isolation of canid herpesvirus 1 in Argentina / Uma apresentação clínica atípica para o primeiro isolamento do herpesvirus canino 1 na Argentina

Relatam-se o primeiro isolamento de herpesvirus canino 1 (CaHV-1) e a localização atípica das lesões vesiculares associadas a este vírus na Argentina. A amostra foi recuperada de lesões vesiculares, localizadas na parte interna da coxa direita, em uma fêmea de raça Labrador. A cadela tinha quatro anos de idade e era de propriedade privada. O primeiro diagnóstico foi realizado pela reação em cadeia da polimerase e, posteriormente, o vírus foi isolado e sua identificação confirmada por imunofluorescência indireta e pelo teste de neutralização viral.

Embryo-toxic effects of environmental concentrations of chlorpyrifos on the crustacean Daphnia magna.

Chlorpyrifos is a non-systemic organophosphorus insecticide leading to the inhibition of the enzyme cholinesterase. Nowadays, this insecticide is widely used on a variety of crops and is frequently detected in surface waters around the world. The main aims of this study were to evaluate the effects of this insecticide on the embryonic development of Daphnia magna, and to assess the sensitivity of this endpoint in comparison with the endpoint of the chronic 21-day test. The featured endpoints were the number of offspring per female and abnormalities of both offspring and embryos. Chlorpyrifos exhibited significant toxic effects on the embryonic development manifested in the appearance of abnormalities, including arrested eggs, in 20% of the embryos exposed to a concentration of 0.01 microgL(-1). Furthermore, chronic exposition to the insecticide yielded a proportion of offspring abnormalities in later stages of development of about 43% at 0.01 microgL(-1). In contrast, a significant reduction in the number of offspring produced per female was only observed above a concentration of 0.03 microgL(-1). Hence, these results suggest that developmental abnormality was a more sensitive endpoint than the number of offspring production per female.

Assessment of the pesticides atrazine, endosulfan sulphate and chlorpyrifos for juvenoid-related endocrine activity using Daphnia magna.

The water flea Daphnia magna belongs to the cyclical parthenogenic species, which can reproduce by either parthenogenesis or sexual reproduction. Recent studies have reported the involvement of the methylfarnesoate hormone, in male sex determination of D. magna. The aim of this study was to evaluate the juvenoid and anti-juvenoid activity of atrazine, endosulfan sulphate, and chlorpyrifos. To assess the juvenoid activity we exposed maternal daphnids to several concentrations of the respective pesticides, using the percentage of male production as endpoint. Fenoxycarb (1microgL(-1)) was used as a positive control. The anti-juvenoid activity was assessed using a similar bioassay after the addition of fenoxycarb (1microgL(-1)) to all test solutions. Fenoxycarb is an insect growth regulator that mimics the action of methylfarnesoate, and promoted the production of 95% of male offspring at the given concentration. Weak juvenoid-agonist activity was detected for endosulfan sulphate, with a significant increase of the percentage of male daphnids detected through logistic regression. In addition, atrazine and endosulfan sulphate antagonized the juvenoid activity of fenoxycarb. These results also corroborate the hypothesis that weak juvenoid agonists can simultaneously act as juvenoid antagonists as it has been observed in other hormonal systems.

A step forwards in ethanol selective photo-oxidation.

This work focuses on the optimization of a photocatalyst formulation for the selective oxidation of ethanol to acetaldehyde. VO(x)/TiO(2) catalysts at different vanadium loading were studied in a gas-solid photocatalytic fluidized bed reactor at high illumination efficiency, in which the bed is constituted by photocatalyst diluted with alpha-Al(2)O(3) or silica gel. Photocatalytic tests showed the selective formation of acetaldehyde, with ethylene and carbon dioxide as by-products. Selectivity is influenced by the vanadium loading. For ethanol inlet concentration of 0.2 vol%, maximum conversion and acetaldehyde selectivity of 73% and 97%, respectively, were obtained at 100 degrees C on catalyst at nominal 5 wt% V(2)O(5) content (53% of surface monolayer) mixed with alpha-Al(2)O(3). Selective sites were related to surface polymeric vanadates possessing Ti-O-V and V-O-V functionalities while the photoactivity appeared correlated with the catalyst equivalent band gap energy. Increasing the ethanol inlet concentration to 1 vol% and diluting the catalyst with silica gel, total ethanol conversion with about 97% selectivity to acetaldehyde was achieved with a photoreactivity of 0.34 mol m(irradiated)(-3) s(-1). This is three times higher than reported for other photoreactors.

Atrial reservoir function by strain rate imaging in asymptomatic mitral stenosis: prognostic value at 3 year follow-up.

AIMS: Assessment of left and right systolic atrial reservoir function in asymptomatic mitral stenosis (MS) by strain and strain rate imaging (SRI) and their prognostic power at 3 year follow-up was the purpose of this study. There is clear indication to treat (by surgery or percutaneous valvotomy) symptomatic patients with MS, whereas for the asymptomatic ones, the question is much debated. So, we need new echocardiographic parameters helpful for the management of asymtomatic patients. Atrial reservoir function by SRI could help in evaluation of these patients. METHODS AND RESULTS: Fifty-three asymptomatic patients with MS and 53 healthy controls were evaluated by the standard echo-Doppler study [mitral valve (MV) area, mean gradient, systolic pulmonary pressure, left atrial (LA) width, LA volumes, LA compliance index] and by Doppler myocardial imaging (velocity, strain, and SR of both atria). The endpoint at 3 year follow-up was symptoms, hospitalization for cardiac cause, atrial fibrillation, thrombo-embolic events, valvular surgery, or percutaneous commissurotomy. LA width, volumes, and systolic pulmonary pressure were significantly increased in MS patients (P < 0.001). Atrial myocardial velocities and deformation indices were significantly compromised in MS patients (P < 0.0001). Significant correlation was found between atrial myocardial velocity and MV area (by pressure half-time method: P = 0.019, R = 0.41; by planimetric method: P = 0.016, R = 0.43). Peak systolic LA myocardial strain and SR were significantly correlated with atrial volumes (strain: P = 0.03, R = -0.28; SR: P = 0.0008, R = -0.42), with atrial compliance index (strain: P = 0.04, R = 0.26; SR: P = 0.04, R = 0.16), with atrial ejection fraction (strain: P < 0.0001, R = 0.56; SR: P = 0.03, R = 0.43). At 3 year follow-up, 22 (41%) patients had events. Comparing the MS patients who had events during the 3 year follow-up with those who did not, the former had bigger LA volumes, although these parameters did not reached a significant value, whereas atrial myocardial systolic SR was significantly impaired in patients with events. In multivariate analysis, the best predictor of adverse events was LA peak systolic SR average (P = 0.04; coefficient: 0.113; SE: 0.055; cut-off value of 1.69 s(-1) for LA peak systolic SR average) with a sensitivity of 88%, specificity of 80.6%, area under the receiver operating characteristic curve of 0.852 (SE: 0.048; 95% CI: 0.74-0.93, P = 0.0001). CONCLUSION: Atrial myocardial deformation properties, assessed by SRI, are abnormal in asymptomatic patients with rheumatic MS. The degree of this impairment is predictor of events in a 3 year follow-up. SRI could be helpful in decision-making of asymtomatic patients with MS.

Risk assessment of representative and priority pesticides, in surface water of the Alqueva reservoir (South of Portugal) using on-line solid phase extraction-liquid chromatography-tandem mass spectrometry.

Surface waters located in intensive agricultural areas are more vulnerable to the pesticides contamination, which is a major concern if the water is intended to be used for human consumption. The aim of this study was to evaluate the presence and the distribution of pesticides in the Alqueva reservoir, an important source of water supply (South of Portugal), considering their representativeness in the agricultural practice of the area. For the analysis of pesticides risk impact we used the environmental quality standards in the field of water policy proposed recently by the European Commission. The pesticides belonging to the classes of phenylureas, triazines, chloroacetanilides, organophosphorous and thiocarbamates were analysed by on-line solid phase extraction-liquid chromatography-tandem mass spectrometry. The pesticides more frequently detected were atrazine, simazine, diuron and terbuthylazine. The highest levels of these pesticides were registered in spring, after pesticides treatment, namely in olive-tree and vine crops. The priority pesticides atrazine and diuron reached values above the annual average proposed in the European Union Legislation. The herbicide atrazine reached values that surpassed the proposed maximum allowable concentration (2,000 ng L(-1)). The sampling stations most affected by these pesticides were Sra. Ajuda, Lucefecit and Alcarrache, located in the northern part of the reservoir, closer to Spain where the agricultural activity is more intensive.

Endosulfan sulphate interferes with reproduction, embryonic development and sex differentiation in Daphnia magna.

Endosulfan sulphate is the transformation product of endosulfan and it is the most frequent form of surface water contamination with endosulfan. The aim of this study was to evaluate the possible effects promoted by endosulfan sulphate in changes on the life cycle, embryo development and sex differentiation of Daphnia magna. The endpoints used were moulting frequency, fecundity, growth, developmentally abnormalities and sexual differentiation. The nominal concentrations of endosulfan sulphate tested ranged from 9.2 to 458.7 microg L(-1). Endosulfan sulphate promoted a significant decrease of the offspring number in all concentrations. Results showed a reduction of the size of females, together with a decrease in moulting frequency. Furthermore, an increase in embryo deformities was observed at all concentrations tested. Above a concentration of 91.7 microg L(-1) there was an increased production of males. The results suggest that endosulfan sulphate interferes with the life cycle and sex determination of the crustacean D. magna.

Effects of atrazine and endosulfan sulphate on the ecdysteroid system of Daphnia magna.

The ecdysteroid system is used by crustaceans and other arthropods as the major endocrine signalling molecules, regulating processes such as molting and embryonic development. The aim of this study was to evaluate the ecdysteroid activity of two pesticides (atrazine and endosulfan sulphate), with distinct modes of action and which act in the juvenoid system of the crustacean Daphnia magna as weak juvenoid compounds. To assess the ecdysteroid activity, we first exposed maternal daphnids and embryos to nominal concentrations of the pesticides and determined the effect promoted by these pesticides on the molting frequency and on abnormalities in the embryos development. Furthermore, we evaluated if the toxic effects observed with the isolated pesticides were promoted or not by the disruption of the ecdysteroids system of the crustacean, by co-administrating of them with 20-hydroxyecdysone hormone. This hormone is the prime form of the invertebrate's ecdysteroids system and it is responsible for the crustacean molting process. Both pesticides induced an increase of embryo abnormalities development. Endosulfan sulphate promoted a delay in the molting process. The effects induced by atrazine were not altered by co-exposure to 20-hydroxyecdysone. In contrast, the co-administration of 20-hydroxyecdysone allowed the reversion of the effects on both the molting process and embryonic development elicited by endosulfan sulphate. These results suggest that atrazine promotes its toxicity without interfering with the ecdysteroid activity of the crustacean. On the contrary, endosulfan sulphate is an anti-ecdysteroidal compound for D. magna.

Acute toxicity of atrazine, endosulfan sulphate and chlorpyrifos to Vibrio fischeri, Thamnocephalus platyurus and Daphnia magna, relative to their concentrations in surface waters from the Alentejo region of Portugal.

Ecotoxicological effects of the herbicide atrazine and the insecticides endosulfan sulphate and chlorpyrifos were evaluated using a test battery comprising aquatic organisms from different trophic levels. According to the categories established in the EU legislation, atrazine can be considered non-harmful for the species tested, while the insecticides can be considered very toxic for the crustaceans. The results of acute toxicity tests showed that the sensitivity of organisms were as follows: Thamnocephalus platyurus > Daphnia magna > Vibrio fischeri. Chlorpyrifos may act as a toxic compound in the aquatic environment of Guadiana River, as it may be detected in water at levels that promote toxic effects.

Impact of malolactic fermentation on low molecular weight phenolic compounds.

A reversed phase liquid chromatography-DAD method is proposed for analysis of major non-flavonoid phenolic compounds in wines. The method employed a mixture of acetic acid, water and methanol as eluents and was used to evaluate the impact of malolactic fermentation in low molecular phenolic compounds. The wines analyzed underwent different treatments, like the addition of a pectolytic enzyme or lysozyme, and the way malolactic fermentation was carried out-spontaneously or with the inoculation of two different commercial lactic bacteria. The main result observed was the disappearance of hydroxycinnamoyltartaric acids and the increase of resultant free forms, regardless the way malolactic fermentation was carried out.

[Hierarchical clustering analysis to detect associations between clinical and pathological features of gastric tumors and hypermethylation of suppressor genes]. / Identificación de asociaciones clínico-patológicas e hipermetilación de genes supresores de tumores en cáncer gástrico difuso a través de análisis de Hierarchical clustering.

BACKGROUND: Methylation is an inactivation mechanism for tumor suppressor genes, that can have important clinical implications. AIM: To analyze the methylation status of 11 tumor suppressor genes in pathological samples of diffuse gastric cancer. MATERIAL AND METHODS: Eighty three patients with diffuse gastric cancer with information about survival and infection with Epstein Barr virus, were studied. DNA was extracted from pathological slides and the methylation status of genes p14, p15, p16, APC, p73, FHIT, E-cadherin, SEMA3B, BRCA-1, MINT-2 y MGMT, was studied using sodium bisulphite modification and polymerase chain reaction. Results were grouped according to the methylation index or Hierarchical clustering (TIGR MultiExperiment Viewer). RESULTS: Three genes had a high frequency of methylation (FHIT, BRCA1, APC), four had an intermediate frequency (p15, MGMT, p14, MINT2) and four had a low frequency (p16, p73, E-cadherin, SEMA3B). The methylation index had no association with clinical or pathological features of tumors or patients survival. Hierarchical clustering generated two clusters. One grouped clinical and pathological features with FHIT, BRCA1, and APC and the other grouped the other eight genes and Epstein Barr virus infection. Two significant associations were found, between APC and survival and p16/p14 and Epstein Barr virus infection. CONCLUSIONS: Hierarchical clustering is a tool that identifies associations between clinical and pathological features of tumors and methylation of tumor suppressor genes.