Development Of An Oxidative Cytomic Panel Of High-Content Miniaturized Assays for Detection of In Vitro Cytotoxicity and Prediction of Acute Toxicity of Drugs and Xenobiotics

La predicción de la toxicidad de xenobióticos (cosméticos, productos químicos y medicamentos ) en seres humanos es esencial en la evaluación de riesgos y la Toxicología reguladora. En la actualidad, las nuevas directrices europeas instan a probar estrategias novedosas para evaluar y validar la seguridad de los medicamentos, productos químicos y cosméticos. Consideraciones éticas y económicas exigen métodos in vitro validados como una alternativa al uso de animales para la detección y predicción de toxicidad humana de acuerdo con el principio de las 3Rs (reemplazo, reducción y refinamiento). Para aumentar la capacidad de predicción de la toxicidad aguda en humanos mediante la cuantificación in vitro de ensayos funcionales, nos hemos planteado validar un grupo de ensayos citómicos miniaturizados para la detección de estrés oxidativo en tres líneas celulares de origen humano: hepatoma (HepG2), neuroblastoma (SH-SY5Y) y adenocarcinoma de riñón (A-704). Se han seleccionado 60 compuestos (Tabla 2), que incluyen fármacos y productos químicos, para los cuales se ha documentado la toxicidad aguda in vivo (LD50 o LC50 en sangre), con datos evaluados por el proyecto europeo ACuteTox y depositados en la base de datos Acutoxbase (Sjöström et al., 2008), además de su citotoxicidad basal in vitro, determinada por el ensayo de captación de Rojo Neutro en células 3T3, validado por ICCVAM/ECVAM. Dentro del conjunto de tóxicos seleccionados para nuestro estudio se incluyen, por una parte, compuestos en los que los ensayos de citotoxicidad humana basal predicen la toxicidad humana aguda y compuestos que no se localizan sobre la recta de regresión en la correlación in vitro/in vivo (“outliers”). Las determinaciones de citotoxicidad por CMF incluyen la cuantificación de la actividad peroxidativa, de los niveles intracelulares de anión superóxido y de óxido nítrico y las determinaciones por HCA de los niveles de la base modificada 8-oxo como indicadores de lesión oxidativa al DNA. Posteriormente se utilizó una línea celular con capacidad metabólica (HepaRG) y se comparó los resultados obtenidos con los valores de citotoxicidad basal de HepG2. Finalmente se determinó la presencia de transportadores de fármacos en la membrana plasmática de las cuatro líneas celulares por citometría de flujo y qPCR además de verificar su funcionalidad mediante ensayos de eflujo de sustratos fluorescentes. El objetivo principal del estudio es mejorar el valor predictivo de la citoxicidad basal como alternativa in vitro a los animales de laboratorio, mediante la puesta a punto de procedimientos miniaturizados de cultivo celular y la incorporación de parámetros más específicos, indicadores de dianas moleculares. Esto se abordará mediante la investigación de las alteraciones celulares tempranas inducidas por compuestos tóxicos, aplicando una plataforma de ensayos de toxicidad de alto contenido, el conjunto de ensayos denominados "Panel Citómico de Ensayos de Estrés Oxidativo”. Los objetivos concretos son: 1 – Miniaturizar y optimizar un Panel Citómico de Ensayos de Estrés Oxidativo basado en la citometría de flujo para el análisis in vitro en células humanas susceptible de ser adaptado a la robotización y al análisis bioinformático de los resultados. 2 – Evaluar la capacidad predictiva de toxicidad humana aguda in vivo del Panel Citómico de Ensayos de Estrés Oxidativo mediante su aplicación a una serie de compuestos en los que la correlación entre toxicidad in vivo e in vitro previamente investigada en estudios multicéntricos. 3 – Validar el Panel Citómico de Ensayos de Estrés Oxidativo mediante la correlación de los valores de citotoxicidad obtenidos en los ensayos de estrés oxidativo con los obtenidos en ensayos in vitro de referencia. 4 – Determinar la capacidad del Panel Citómico de Ensayos de Estrés Oxidativo para clasificar correctamente los compuestos usando el sistema GHS adaptado a los datos de humano. 5 – Determinar la capacidad de estos ensayos para predecir la toxicidad órgano-específica. 6 – Aplicar el Panel Citómico de Ensayos de Estrés Oxidativo al análisis cuantitativo in vitro de la citotoxicidad basal y la toxicidad dependiente de biotransformación de diferentes xenobióticos. 7 – Determinar mediante citometría de flujo, PCR cuantitativa y ensayos funcionales la expresión de transportadores multifármaco en las líneas utilizadas.Prediction of toxicity of xenobiotics to humans is essential for risk assessment and regulatory Toxicology. Currently, the new European directives urge novel test strategies to be validated for assessing the safety of drugs, chemicals and cosmetics. Ethical and economic considerations, as well as regulatory imperatives to protect both consumers and animals, demand in vitro methods, as validated alternatives to laboratory animals for the detection and prediction of human toxicity, according to the 3Rs principle (Replacement, Reduction and Refinement). In the European Integrated Project “A-Cute-Tox” our laboratory played a leading role to explore new cell systems and new endpoints for in vitro cytotoxicity. In our work with human cell lines, we have found two essential drawbacks in many cell lines extensively used: the presence of mechanisms of drug efflux and the loss of biotransformation activity in liver-derived cells. One limitation of in vitro methods to predict accurately in vivo toxicity is the overexpression in many cells lines of drug resistance mechanisms. The most studied factor of drug resistance is MDR1 gene and its product P-glycoprotein (P-gp), an ATP-dependent extraction pump. P-gp is expressed in normal tissues linked to secretion or barrier functions and in tumors confers the multidrug resistance phenotype. P-gp is overexpressed in renal adenocarcinoma, hepatoma and neuroblastoma (4), tumor types originating the cell lines most used for in vitro studies of nephrotoxicity, hepatotoxicity and neurotoxicity. However, no systematic studies exist on the quantitative influence of efflux pumps on in vitro cytotoxicity data and their extrapolation to human in vivo toxicity. The use of cell lines to predict in vivo toxicity is limited in studies of biotransformation-dependent toxicity. Human primary hepatocytes and immortalized hepatocytes are good but limited alternatives. Primary hepatocytes are scarcely available, have limited growth and lifespan, and undergo early phenotypic alterations. Immortalized hepatocytes loss most of their biotransformation activities. The novel hepatocyte-like cell Hepa-RG (human hepatoma cell line derived from hepatocellular carcinoma) is a promising alternative, since express hepatocyte markers, have various secretory and metabolic functions and drug detoxification activities Usually, in vitro cytotoxicity is estimated as cell death rate, while qualitative studies of intracellular biochemistry are only designed for mechanistic toxicology. Also, most studies involve bulk measurements rather than attempting single-cell based determinations. Flow cytometry and high-content analysis by bioimage (HCA) are based on single-cell multiparametric fluorescence measurements of heterogeneous cell populations while maintaining a typical acquisition rate of hundreds-thousands of cells per second. These novel cytomic functional assays detect early or transient events, being advantageous over assays that quantify simple endpoints, as cell death or general cellular alterations. Oxidative stress, a consequence of excessive production of reactive oxygen species or decreased antioxidant defense, causes cytotoxicity through structural and functional alterations with disruption of cellular homeostasis. To increase the predictivity of acute human toxicity by quantitative in vitro functional assays, we will validate a Cytomic Assay Panel for Oxidative Stress Screening using biotransformation-competent cell lines. The main objective is to improve the predictive value of basal cytotoxicity and biotransformation-dependent cytotoxicity assays as in vitro alternatives to laboratory animals. This will be achieved by investigating the early cellular alterations induced by relevant toxic compounds through a novel toxicity high-content screening strategy (Cytomic Panel of Oxidative Stress Assays) . The specific objectives of the project are: 1. To miniaturize and optimize a panel of cytomic high content assays of oxidative stress based upon automated flow cytometry and HCA by Bioimage in human cell lines that may be amenable to robotization and bioinformatic analysis. 2. To assess the predictive capacity for human acute toxicity of the cytomic assay panel by means of its application to a subset of chemicals whose correlation between in vitro and in vivo toxicities has been previously reported by multicentric studies. 3. To validate the assay panel through the correlation between the cytotoxicity values generated with the assay panel and those obtained in parallel using accepted assays of in vitro cytotoxicity (Neutral Red uptake, MTT, LDH release). 4. To determine the capacity to predict organ specific toxicity of the cytomic assay panel. 5. To determine the ability to correctly classify the compounds using a system for chemical toxicity classification in humans, adapted from the Global Harmonization System (GHS) of the Cytomic Panel of Oxidative Stress panel. 6. To apply this assay panel to quantitative in vitro analysis of basal and biotransformation-dependent cytotoxicity of different reference toxic compounds. 7. To determine the expression of multidrug resistence proteins in the cells

Comportamento em condições de campo de cafeeiros (Coffea arabica L.) propagados vegetativamente e por semeadura

Cutting is an alternative method for vegetative propagation of Coffea arabica L. hybrids for commercial purpose. However, the use of such method requires the understanding of the plant growth characteristics under filed conditions. Thus, the objective of the present study was to evaluate and compare the vegetative growth and first yield of Coffea arabica cv. Acaiá plants obtained by cuttings and with seed propagated plants. The experiment was installed in the year 2003 and carried out in the Department of Agriculture at Federal University of Lavras-UFLA. For cutting propagation, stems were treated with three concentrations (0, 2000 e 4000 mg.L-1) of Indol butiric acid (AIB), in the presence or absence of supplemental warming. The development and growth of the cuttings were compared to the plantlets obtained by traditional seed propagation. A randomized blocks design, with six plants per plot, considering four useful plants and three replications was used. The vegetative growth, height, and yield of the plants of coffee obtained from cutting and treated with 2000 mg.L-1 AIB, were significantly greater than those not treated. Also, the production of plagiotropic branches by the plants originated from cuttings was greater than those originated from seeds.A propagação vegetativa por meio do enraizamento de estacas é uma alternativa para a propagação de híbridos de Coffea arabica L. em escala comercial. Entretanto, para a utilização da propagação via enraizamento de estacas, é necessário o conhecimento das características de crescimento das plantas no campo. Assim, objetivou-se neste trabalho avaliar o crescimento vegetativo e a primeira produção de plantas de Coffea arabica cv. Acaiá, provenientes de estaquia, bem como compará-las com plantas provenientes de semeadura. O experimento foi instalado em 2003 no Departamento de Agricultura da Universidade Federal de Lavras – UFLA. Foram utilizadas mudas provenientes de estaquia, tratadas na fase de enraizamento com ácido indol–butírico (AIB), nas concentrações de (0, 2000 e 4000 mg.L-1) com e sem aquecimento no leito de enraizamento. Como tratamento adicional, foram utilizadas mudas provenientes de semeadura. O delineamento experimental foi em blocos casualizados, com três repetições e parcelas de seis plantas, sendo quatro plantas úteis. Avaliações do crescimento vegetativo e da produção possibilitaram concluir que plantas provenientes de estaquia, tratadas com AIB, apresentaram maior altura e produção em relação às não-tratadas. Observou-se maior número de pares de ramos plagiotrópicos e produção das plantas provenientes de estaquia em relação às plantas provenientes de semeadura

Communication and Biodegradable Packaging Relationship: A Paradigm for Final Disposal

Solid waste generation is estimated to increase from 1.3 billion to 2.2 billion tons by 2025, causing environmental, social, and consequently public health problems. The biggest problem in this regard involves the inadequate disposal of waste, and in emerging countries like Brazil, it is sorted less waste for recycling or composting. In this context, plastic packaging is more complex due to the high polymer composition, as well as low recycling rates. Bioplastics appear as alternatives because they are mostly biodegradable. Given the various functions of packaging and a systematic review of the literature, the aim of this study was to discuss the communicational aspects directly related to bioplastic packaging and to present how the communication function in packaging can contribute to providing relevant information to consumers, to minimize the problem of improper disposal. This paper concluded that communication, whether in plastic or bioplastic packaging should be an agent of environmental education. Thus, promoting essential actions in the people such as non-generation, reduction, reuse, and recycling of waste, consequently, generating a solution cycle that allows the development of a circular economy

External validation of models to predict hepatocellular carcinoma in Hepatitis C Virus cured F3-F4 patients

[EN] Background & AimsSeveral hepatocellular carcinoma (HCC) risk-models have been developed to individualise patient surveillance following sustained viral response (SVR) in Hepatitis C Virus patients. Validation of these models in different cohorts is an important step to incorporate a more personalised risk assessment in clinical practice. We aimed at applying these models to stratify the risk in our patients and potentially determine cost-saving associated with individualised HCC risk-stratification screening strategy. MethodsPatients with baseline F3-4 fibrosis treated with new oral direct-acting antivirals who had reached a SVR were regularly followed as part of the HCC surveillance strategy. Six models were applied: Pons, aMAP, Ioannou, HCC risk, Alonso and Semmler. Validation of the models was performed based on sensitivity and the proportion of patients labelled as "high risk". ResultsAfter excluding 557 with less than 3 fibrosis, 12 without SVR, 18 with a follow up (FU) <1 year, 17 transplant recipients, 16 lost to FU and 31 with HCC at time of antiviral therapy, our cohort consisted of 349 F3-4 SVR patients. Twenty-three patients (6.6%) developed HCC after a median FU of 5.12 years. The sensitivity of the different models varied between 0.17 (Semmler7noalcohol) and 1 (Alonso A and aMAP). The lowest proportion of high-risk patients corresponded to the Semmler-noalcohol model (5%). Sixty-three and 90% of the Alonso A and aMAP patients, respectively were labelled as high risk. The most reliable HCC risk-model applied to our cohort to predict HCC development is the Alonso model (based on fibrosis stage assessed by liver stiffness measurements or Fibrosis-4 index (FIB-4) at baseline and after 1 year, and albumin levels at 1 year) with a-100% sensitivity in detecting HCC among those at high risk and 63% labelled as high risk. The application of the model would have saved the cost of 1290 ultrasound no longer being performed in the 37% low-risk group. ConclusionIn our cohort, the Alonso A model allows the most reliable reduction in HCC screening resulting in safely stopping life-long monitoring in about a third of F3-F4 patients achieving SVR with DAAs.; imageInstituto de Salud Carlos III, Grant/Award Numbers: FI20/00033, INT20/00061, PI19/01360; Centro de Investigacion Biomedica en Red de Enfermedades Hepaticas y Digestivas; Gilead Research Scholars, Grant/Award Number: GLD18-192Carvalho-Gomes, Â.; Valcheva, TV.; Sahuco, I.; Vidal, E.; Martínez-Arenas, L.; Vinaixa, C.; Aguilera, V.... (2024). External validation of models to predict hepatocellular carcinoma in Hepatitis C Virus cured F3-F4 patients. United European Gastroenterology Journal. https://doi.org/10.1002/ueg2.1257

Long-term Negative Psychological Impact of Presymptomatic Testing on Familial Amyloid Polyneuropathy: Efecto psicológico negativo a largo plazo de las pruebas genéticas presintomáticas en la polineuropatía amiloide familiar

http://sherpa.ac.uk/romeo/search.php?issn=2174-0550This study addresses the profile of at-risk subjects whose long-term psychological impact of presymptomatic testing (PST) for Familial Amyloid Polyneuropathy (FAP) TTR V30M is negative. The sample consisted of 177 subjects, aged over 20 years that were 50% at-risk for FAP, and performed the PST at least three years ago. Participants were contacted by mail, one time only, to answer the sociodemographic questionnaire and the Brief Symptom Inventory (BSI), the Self-Rating Anxiety Scale of Zung (SAS), and the Beck Depression Inventory (BDI). From the sample, 22.6% (BSI), 16.4% (SAS), and 9% (BDI) subjects presented negative psychological impact, after having performed the PST for more than 3 years. Subjects with clinically significant values in BSI, SAS, and BDI have an overlapping profile concerning the total sample, except regarding age, since clinically depressed subjects have a higher mean age. Married women or living in unmarried unions, aged between 30 and 45 years, employed, carriers, and having performed the PST test for 6-7 years are a group raising higher concern and requiring a more active role with respect to the psychological impact of the PST for FAP. The role of the clinical and health psychologist with these patients is critical in the adjustment to the presymptomatic test result as well as in adherence to the available treatments conducive to a better quality of life, in carriers.Este estudio aborda el perfil de sujetos en riesgo cuyo impacto psicológico a largo plazo de las pruebas presintomáticas (PST) para la polineuropatía amiloide familiar (FAP) TTR V30M es negativo. La muestra consistió en 177 sujetos mayores de 20 años que tenían un 50% de riesgo de FAP, que habían realizado el PST hacía al menos tres años. Se contactó con los participantes por correo, solo una vez, para responder el cuestionario sociodemográfico y el Inventario de síntomas breves (BSI), la Escala de ansiedad de autoclasificación de Zung (SAS) y el Inventario de depresión de Beck (BDI). El 22.6% (BSI), el 16.4% (SAS) y el 9% (BDI) de los sujetos de la muestra presentaron un impacto psicológico negativo después de haber realizado el PST durante más de 3 años. Los sujetos con valores clínicamente significativos en BSI, SAS y BDI tienen un perfil superpuesto con respecto a la muestra total, excepto con respecto a la edad, ya que los sujetos clínicamente deprimidos tienen una edad media más alta. Las mujeres casadas o que viven en pareja, con edades entre 30 y 45 años, que trabajan, son portadoras y han realizado la prueba PST durante 6-7 años son un grupo que suscita una mayor preocupación y requiere un papel más activo con respecto al impacto psicológico del PST para FAP. El papel del psicólogo clínico y de la salud con estos pacientes es decisivo en el ajuste del resultado de la prueba presintomática, así como en la adhesión a los tratamientos disponibles que conducen a una mejor calidad de vida en los portadores.info:eu-repo/semantics/publishedVersio

Study of anatomical variations in premolars by cone beam computerized tomography in a radiologic clinic in Piauí

Introduction: Root canal cleaning is the main objective of endodontic treatment and requires knowledge of the internal anatomy. The premolars are evidenced in the literature with great anatomical variations. In view of this, studies indicate that the use of Cone Beam Computed Tomography helps in the visualization of highly complex anatomy. Objective: to describe the anatomical variations in maxillary and mandibular premolars using cone beam computed tomography in a radiologic clinic in Piaui. Methods: 54 cone beam computed tomography scans with 160 premolars were used, produced using the Orthopantomograph OP300 equipment and analyzed by multiplanar reconstructions: axial, coronal and sagittal. Data regarding sex, number of roots and canals were recorded to compare and classify according to Vertucci. Results: the maxillary first pre-molars had 63.5% two roots,83.7% with one root and the mandibular pre-molars mostly with one root. Regarding the number of channels, 92.3% of the first premolars had two channels, most of them maxillary second premolars and mandibular premolars only one channel. Vertucci variations of types I, II, III and IV were verified in single-rooted elements, observing a great variation in superior elements. As for the prevalence of sex, only the first superiors showed greater variation in males. Conclusions: the upper first premolars prevailed with a great anatomical variation in relation to the other premolars with prevalence of Vertucci Type I and in males.Introducción: La limpieza del conducto radicular es el principal objetivo del tratamiento endodóntico y requiere del conocimiento de la anatomía interna. Los premolares se evidencian en la literatura con grandes variaciones anatómicas. Por lo tanto, los estudios indican que el uso de la tomografía computarizada de haz cónico ayuda en la visualización de anatomías altamente complejas. Metodología: Se utilizaron 4 equipos computarizados 60 premolares con 1 equipo, equipo sin equipo Orthopanto y ahora por reconstrucciones, coronal y sargital. Se registraron datos referentes al sexo, número de raíces y conductos para su comparación y clasificación según Vers. Resultados: Los primeros molares maxilares tenían el 63,5% de dos raíces, el 83,7% de los segundos premolares superiores tenían una raíz y la mayoría de los premolares mandibulares tenían una raíz. En cuanto al número de canales, el 92,3% de los primeros premolares tenían dos canales, siendo la mayoría segundos premolares maxilares y premolares inferiores de un canal. Se verificaron variaciones de Vertucci de los tipos I, II, III y IV en los elementos monoraíces, observándose la gran observación en los elementos superiores. En cuanto a la mayoría de machos, los primeros presentan el mayor número de machos. Conclusión: Los primeros premolares superiores prevalecerán con los demás premolares en relación a la prevalencia de Vertucci y en el sexo masculino.Introduction : Le nettoyage canalaire est l'objectif principal du traitement endodontique et nécessite une connaissance de l'anatomie interne. Les prémolaires sont mises en évidence dans la littérature avec de grandes variations anatomiques. Par conséquent, des études indiquent que l'utilisation de la tomodensitométrie à faisceau conique aide à la visualisation d'anatomies très complexes. Méthodologie : 4 équipements informatisés ont été utilisés 60 prémolaires avec 1 équipement, équipement sans équipement Orthopanto et maintenant par reconstructions, coronales et sarcitales. Les données relatives au sexe, au nombre de racines et de canaux ont été enregistrées pour comparaison et classification selon Vers. Résultats : Les premières molaires maxillaires avaient 63,5 % de deux racines, 83,7 % des deuxièmes prémolaires maxillaires avaient une racine et la plupart des prémolaires mandibulaires avaient une racine. En ce qui concerne le nombre de canaux, 92,3% des premières prémolaires avaient deux canaux, la majorité n'étant que des deuxièmes prémolaires maxillaires et des prémolaires inférieures à un seul canal. Les variations de Vertucci des types I, II, III et IV ont été vérifiées dans les éléments à racine unique, en observant la grande observation dans les éléments supérieurs. Quant à la majorité des mâles, les premiers à présenter le plus grand nombre de mâles. Conclusion&nbsp;: Les premières prémolaires supérieures prédominent avec les autres prémolaires en relation avec la prévalence de Vertucci et chez les hommes.Introdução: a limpeza do canal radicular é o principal objetivo do tratamento endodôntico e requer conhecimento da anatomia interna. Os pré-molares são evidenciados na literatura com grandes variações anatômicas. Diante disso, estudos indicam que o uso da Tomografia Computadorizada Cone Beam auxilia na visualização de anatomias de alta complexidade. Metodologia: foram utilizadas 54 tomografias computadorizadas de feixe cônico com 160 pré-molares, produzidas no equipamento Orthopantomograph OP300 e analisadas por reconstruções multiplanares: axial, coronal e sagital. Os dados referentes ao sexo, número de raízes e canais foram registrados para comparação e classificação segundo Vertucci. Resultados: os primeiros pré-molares superiores apresentavam 63,5% de duas raízes, 83,7% dos segundos pré-molares superiores tinham uma raiz e a maioria dos pré-molares inferiores tinha uma raiz. Em relação ao número de canais, 92,3% dos primeiros pré-molares possuíam dois canais, sendo a maioria segundos pré-molares superiores e pré-molares inferiores apenas um canal. Vertucci variações dos tipos I, II, III e IV foram verificadas nos elementos uniradiculares, observando-se a grande variação nos elementos superiores. Quanto à prevalência do sexo, apenas os primeiros superiores apresentaram maior variação no sexo masculino. Conclusão: os primeiros pré-molares superiores prevaleceram com grande variação anatômica em relação aos demais pré-molares com prevalência de Vertucci Tipo I e no sexo masculino

Long-term negative Psychological Impact of Presymptomatic Testing for Huntington’s Disease

http://sherpa.ac.uk/romeo/issn/2422-8419/Presymptomatic Testing (PST) for Huntington’s disease (HD) is available since 1986 and its impact on carriers and non-carriers is not yet fully clear. It is important to understand its psychological impact so that the PST protocols are best suited to the subjects at-risk. Preventing a negative psychological impact is the ultimate purpose of the genetic counselling process. This study addresses the long-term negative psychological impact assessment of PST for HD. The sample consisted of 29 subjects that were 50% at-risk for HD, and had performed the PST for at least three years ago. Participants answered the sociodemographic questionnaire and the Brief Symptom Inventory, the Zung Self-Rating Anxiety Scale, and the Beck Depression Inventory. Although most of the sample does not present clinically significant psychopathology values, 6 subjects present a Positive Symptoms Distress Index value which is equal to or greater than 1.7; 7 subjects present a value which is equal to or greater than 40 of anxiety; and 7 subjects present mild depression. Symptomatic carriers, who underwent the PST less time ago, present worse psychopathological symptoms, depression and anxiety. Subjects with this profile should have a more intense and personalized psychological and social support, aiming to prevent the risk of suicide and to improve the quality of their lives.info:eu-repo/semantics/publishedVersio

Long-term negative Psychological Impact of Presymptomatic Testing for Huntington’s Disease

http://sherpa.ac.uk/romeo/issn/2422-8419/Presymptomatic Testing (PST) for Huntington’s disease (HD) is available since 1986 and its impact on carriers and non-carriers is not yet fully clear. It is important to understand its psychological impact so that the PST protocols are best suited to the subjects at-risk. Preventing a negative psychological impact is the ultimate purpose of the genetic counselling process. This study addresses the long-term negative psychological impact assessment of PST for HD. The sample consisted of 29 subjects that were 50% at-risk for HD, and had performed the PST for at least three years ago. Participants answered the sociodemographic questionnaire and the Brief Symptom Inventory, the Zung Self-Rating Anxiety Scale, and the Beck Depression Inventory. Although most of the sample does not present clinically significant psychopathology values, 6 subjects present a Positive Symptoms Distress Index value which is equal to or greater than 1.7; 7 subjects present a value which is equal to or greater than 40 of anxiety; and 7 subjects present mild depression. Symptomatic carriers, who underwent the PST less time ago, present worse psychopathological symptoms, depression and anxiety. Subjects with this profile should have a more intense and personalized psychological and social support, aiming to prevent the risk of suicide and to improve the quality of their lives.info:eu-repo/semantics/publishedVersio

Long-term Negative Psychological Impact of Presymptomatic Testing for Huntington ’s disease

Presymptomatic Testing (PST) for Huntington’s disease (HD) is available since 1986 and its impact on carriers and non-carriers is not yet fully clear. It is important to understand its psychological impact so that the PST protocols are best suited to the subjects at-risk. Preventing a negative psychological impact is the ultimate purpose of the genetic counselling process. This study addresses the long-term negative psychological impact assessment of PST for HD. The sample consisted of 29 subjects that were 50% at-risk for HD, and had performed the PST for at least three years ago. Participants answered the sociodemographic questionnaire and the Brief Symptom Inventory, the Zung Self-Rating Anxiety Scale, and the Beck Depression Inventory. Although most of the sample does not present clinically significant psychopathology values, 6 subjects present a Positive Symptoms Distress Index value which is equal to or greater than 1.7; 7 subjects present a value which is equal to or greater than 40 of anxiety; and 7 subjects present mild depression. Symptomatic carriers, who underwent the PST less time ago, present worse psychopathological symptoms, depression and anxiety. Subjects with this profile should have a more intense and personalized psychological and social support, aiming to prevent the risk of suicide and to improve the quality of their lives. Keywords: Presymptomatic Testing (PST), Preditive genetic testing, Huntington´s Disease (HD), Long-term Psychological Impact, Late-onset diseas

Cd-tolerance markers of Pfaffia glomerata (Spreng.) Pedersen plants: Anatomical and physiological features

Physiological and anatomical features of Cd-tolerance in Pfaffia glomerata were examined by exposing plantlets to nutrient solutions with increasing Cd concentrations (0, 15, 45, and 90 μmol Cd L-1), and possible Cd-tolerance markers were established. Cd contents were found to be higher in roots than in shoots. According to the bio-concentration factor data, this species is effectively a Cd-hyperaccumulator, as previously attested. Cd induced the appearance of xeromorphic characteristics in leaves (decreased water potential, increased numbers and decreased stomata size) and increased root endodermis thickness. The enzymatic antioxidant systems of roots and leaves were differently affected by Cd. The coordinated activities of antioxidant enzymes were effective in reducing Cd-induced reactive oxygen species in plants, mainly in leaves. Root endodermis thickness, stomatal size and numbers, root superoxide dismutase, and guaiacol peroxidase, as well as leaf guaiacol peroxidase and catalase activities can all be considered Cd-tolerance markers in Pfaffia glomerata. Due to its high root Cd accumulation, Pfaffia glomerata may be useful in Cd-phytoextraction programs, however the pharmacological use of plants grown in the presence of Cd must be avoided