The effects of a plant proteinase inhibitor from Enterolobium contortisiliquum on human tumor cell lines

Supplementary to the efficient inhibition of trypsin, chymotrypsin, plasma kallikrein, and plasmin already described by the EcTI inhibitor from Enterolobium contortisiliquum, it also blocks human neutrophil elastase (K(iapp)=4.3 nM) and prevents phorbol ester (PMA)-stimulated activation of matrix metalloproteinase (MMP)-2 probably via interference with membrane-type 1 (MT1)-MMP. Moreover, plasminogen-induced activation of proMMP-9 and processing of active MMP-2 was also inhibited. Furthermore, the effect of EcTI on the human cancer cell lines HCT116 and HT29 (colorectal), SkBr-3 and MCF-7 (breast), K562 and THP-1 (leukemia), as well as on human primary fibroblasts and human mesenchymal stem cells (hMSCs) was studied. EcTI inhibited in a concentration range of 1.0-2.5 mu M rather specifically tumor cell viability without targeting primary fibroblasts and hMSCs. Taken together, our data indicate that the polyspecific proteinase inhibitor EcTI prevents proMMP activation and is cytotoxic against tumor cells without affecting normal tissue remodeling fibroblasts or regenerative hMSCs being an important tool in the studies of tumor cell development and dissemination

Anthropogenic Disturbances and the Emergence of Native Diseases: a Threat to Forest Health

The next century will almost certainly see an unprecedented rise in forest pathogen epidemics, requiring a proactive rather than reactive response. Diseases caused by native pathogens with complex aetiologies will become more common, and recognising, characterising and managing these epidemics are difficult because native pathogens are frequently already widespread, and eradication is not feasible. We need to start approaching these issues from a ‘whole ecosystem’ perspective, highlighting the many aspects and entanglements of forest declines and allowing us to respond with management options tailored to each scenario. The approach proposed here provides logical steps based on six questions to untangle the direct and indirect environmental drivers of tree declines

Identification of a mosaic non-inherited small supernumerary ring chromosome 2: cytogenetic-molecular studies and genotype-phenotype correlation

Introduction: The identification of supernumerary marker chromosomes (SMCs) derived from all the autosomes is currently possible, but rarely by conventional cytogenetics alone. Supernumerary ring chromosomes (SRCs) account for about 10% of these cases. SRCs derived from chromosome 2 are unusual, and there are only a few cases reported in the literature. The severity of the phenotype depends on the type of the mosaicism, the percentage of cells affected by the genetic change and the chromosome involved. Methods: The authors report the case of a boy aged 8 referred for cytogenetic studies, presenting with behavior and learning problems, mental retardation with uncoordenated speech, attention deficit and hyperactivity (PHDA), as well as small slanting palpebral fissures. The karyotype was obtained from peripheral blood lymphocyte cultures using high resolution GTL banding and standard techniques. Fluorescence in situ hybridization (FISH) was performed using specific probes for the centromeric regions of all chromosomes (Chromoprobe Multiprobe - ISystem). Results: Cytogenetic analysis revealed two cell lines: one with a supernumerary marker ring chromosome, 47,XY,+r (52%), and a normal cell line, 46,XY (48%). The SRC was identified by FISH with the chromosome 2 centromeric probe. Since the parents had normal karyotypes, this abnormality was “de novo”. Final karyotype of the proband was: mos 47,XY,+r[26]/46,XY[24].ish r(2)(D2Z2+)dn. Discussion: The clinical description of this patient is in agreement with other reports of the literature. Molecular characterization by FISH analyses is an useful way of investigating the presence of euchromatin contained in a SMC and establishing new chromosomal syndromes. However, to better characterize this ring, in order to establish a more accurate genotype-phenotype correlation, more studies involving other technologies should be performed, thus allowing suitable genetic counsellin

Deleção cromossómica intersticial em 14q “de novo”: apresentação de um caso

Introdução: As deleções intersticiais são anomalias cromossómicas estruturais, desequilibradas, resultantes de dois pontos de quebra, frequentemente associadas a quadros clínicos anormais devido à perda de material genético ativo (eucromatina). As consequências fenotípicas dependem do segmento cromossómico perdido e do número de genes aí localizados. Material e Métodos: Os autores apresentam o caso de um indivíduo do sexo masculino, de 11 anos de idade, referenciado para estudo citogenético por apresentar um quadro clínico de atraso de desenvolvimento psicomotor, défice cognitivo e problemas de comportamento. Realizaram-se culturas sincronizadas de linfócitos de sangue periférico, bandas GTG de alta resolução e, posteriormente, estudos de hibridação in situ por fluorescência (FISH) com sondas de pintura cromossómica total e subtelomérica, específicas para o cromossoma 14. Resultados: A análise das metafases revelou a presença de uma anomalia estrutural no cromossoma 14, interpretada como uma deleção intersticial do segmento compreendido entre as bandas 14q24.3 e 14q32.1. A análise por FISH permitiu confirmar esta deleção intersticial. Como os cariótipos dos pais foram normais, conclui-se que esta anomalia cromossómica é “de novo”, estabelecendo-se o cariótipo do doente como: 46,XY,del(14)(q24.3q32.1).ish del(14)(wcp 14+,SHGC36156+)dn Discussão: A deleção intersticial encontrada no cromossoma 14 implica uma monossomia do segmento 14q24.3→14q32.1. As alterações descritas mais comuns, associadas a esta deleção, incluem ADPM e algumas malformações minor. Os autores apresentam este caso pela raridade da anomalia citogenética encontrada e comparam-no com a literatura atual

Diagnóstico Pré-natal de Síndrome de Wolf-Hirschhorn: a propósito de um caso

Introdução: O Síndrome de Wolf-Hirschhorn é uma patologia originada por uma deleção da região terminal do braço curto do cromossoma 4. O tamanho da deleção pode ser variável levando a um espectro alargado de manifestações clínicas. Em diagnóstico pré-natal (DPN), as alterações fetais mais frequentes incluem atraso do crescimento intra-uterino, lábio leporino e/ou fenda do palato e anomalias cardíacas. A prevalência estimada é de 1/50.000 nascimentos afetando duas vezes mais indivíduos do sexo feminino do que do sexo masculino. Objectivo: Apresentação de um caso de Síndrome de Wolf-Hirschhorn em DPN comparando-o com outros casos publicados. Material e métodos: Grávida com 17semanas de gestação, referenciada para estudos cromossómicos por idade materna avançada (35 anos) e rastreio bioquímico positivo para trissomia 18. A análise citogenética convencional dos amniócitos cultivados foi realizada de acordo com os métodos habituais usando bandas GTG. O estudo foi complementado por técnicas de citogenética molecular (FISH) utilizando-se a sonda específica para a região do Síndrome de Wolf-Hirschhorn. Resultados: O estudo cromossómico efetuado, revelou uma deleção na região terminal do braço curto do cromossoma 4. A análise por FISH confirmou a existência da deleção desta região, permitindo estabelecer o cariótipo 46,XX,del(4)(p15.3).ish del(4)(p16.3p16.3)(WCHR-). Os cariótipos efetuados aos pais foram normais. Conclusões: Discute-se a importância deste caso pela raridade da anomalia citogenética encontrada, assim como pela dificuldade em realizar o diagnóstico por citogenética convencional, em alguns destes casos, quando não se obtêm bandas de alta resolução

Cystic Fibrosis: actual treatment and future perspectives with nanotechnology

Introducción: Actualmente, los tratamientos existentes para tratar la fibrosis quística (FQ) están diseñados para controlar sus síntomas, consistentes principalmente en retención de moco e infección crónica. Se propone la vía pulmonar como alternativa para la administración de los fármacos, principalmente antimicrobianos. Sin embargo, su rápido aclaramiento, que conduce a niveles bajos de fármaco e incremento de los regímenes posológicos, así como la aparición de efectos adversos, hacen de la nanotecnología una estrategia interesante para esta enfermedad. Objetivo: estudiar y analizar los diferentes sistemas nanoparticulares existentes para su uso por vía pulmonar, concretando en el uso de sistemas lipídicos para el tratamiento de la FQ. Método: se realizó una búsqueda no sistemática de artículos en diferentes bases de datos, en los últimos 10 años principalmente, siguiendo pautas establecidas de palabras clave. Resultados: Los progresos que se han conseguido en los últimos años hacen que la FQ pase a ser una enfermedad de adultos. Los tratamientos que se están usando en la actualidad están siendo cada vez más desplazados por otras alternativas, como los sistemas nanoparticulares, siendo idóneos para la administración pulmonar debido a su pequeño tamaño, su liberación sostenida y su elevada biocompatibilidad. Entre éstos, destacan los liposomas por su similitud estructural con el surfactante pulmonar, así como por su capacidad de destruir las biopelículas bacterianas. La mayoría de las formulaciones encontradas contenían un solo fármaco. Conclusión: Existen evidencias científicas que indican que la investigación debe dirigirse hacia el desarrollo de formulaciones que sean capaces de destruir la biopelícula.Introduction: Currently, the management of treatments in cystic fibrosis (CF) is mainly focused to control symptoms, which consist of mucus retention and chronic infection. The pulmonary route is proposed as an interesting alternative for administering drugs, especially antimicrobials. However, the rapid clearance of these, which leads to low drug levels and increased dosage regimens, as well as the appearance of adverse effects, make nanotechnology an interesting strategy for this disease. Objective: to study and analyze the different nanoparticulate systems available for use via the lung, specifying the use of lipid systems for the treatment of CF. Method: a non-systematic search of articles in different databases was carried out, mainly in the last 10 years, following established guidelines for selecting keywords. Results: The progress in recent years makes CF become an adult disease. Current treatments are increasingly being displaced by other alternatives, such as nanoparticular systems, being suitable for pulmonary administration due to their small size, sustained release and high biocompatibility. Among these, liposomes stand out for their structural similarity to lung surfactant, as well as for their ability to destroy bacterial biofilms. Most of the formulations contained a single drug. Conclusion: Scientific literature evidenced that research studies should be directed towards the development of formulations that are intended to destroy the biofilm

Clinical, cytogenetic and molecular findings of a “de novo” inv dup del (6q)

Introduction: Complex rearrangements resulting in inverted duplications contiguous to a terminal deletion (inv dup del) were first reported for the short arm of chromosome 8 in1976. Since then this type of structural anomaly has been described for an increasing number of chromosomes. In these rearrangements, the concomitant presence of a deletion and a duplication has important consequences in genotype-phenotype correlations. The authors describe the clinical findings and the cytogenetic characterization of a rare inv dup del involving the long arm of chromosome 6. Material and methods: A girl aged 5 was referred for subtelomeric studies with the indication of psychomotor retardation, autistic features and stereotipies. Chromosome analysis with high resolution GTL-banding was performed on metaphases obtained from cultured peripheral blood lymphocytes. Molecular studies included MLPA (Kits P036 and P070, MRC-Holland), FISH with subtelomeric and whole chromosome painting probes specific for chromosome 6, and cCGH techniques. Results: Initial MLPA studies detected a subtelomeric deletion in the long arm of chromosome 6; the subsequent karyotype revealed a structurally abnormal chromosome 6 with additional material in the end of the long arm. FISH analysis showed the deletion and demonstrated that the extra material was derived from chromosome 6; cCGH tecnhiques defined the extension and confirmed the breakpoints of the duplicated segment. Thus this rearrangement was interpreted as an inv dup del (6q). Since parental karyotypes were normal, this anomaly was considered “de novo”. Discussion: As far as we know this is the first description of a patient presenting with a “de novo” inv dup del (6q). We compare the clinical features in this child with the previously reported cases with either an isolated terminal deletion or a duplication of distal 6q. The authors enhance the importance of the combination of high resolution banding with molecular studies in the characterization of this rare rearrangement

Interstitial deletion 15q21 and Prader-Willi like syndrome phenotype: Case report

Introduction: Chromosome 15q interstitial deletions not involving the Prader-Willi/Angelman region are uncommon and poorly characterized. Very few cases of different segmental losses involving the 15q21 region have been reported at cytogenetic level. All the described patients present with moderate to several mental retardation and characteristic facial dysmorphic features. Some authors compare the similarity between the phenotype of these patients with some features of Prader-Willi syndrome (PWS). Methods: We report the case of a girl aged 8 referred for conventional cytogenetics and fluorescence in situ hybridization (FISH) for the PWS region, presenting with mental retardation, almond-shaped eyes, obesity, small hands with short fingers and diminished pigmentation of the hair. Results: The chromosomal analysis revealed an interstitial deletion of the long arm of chromosome 15, apparently between 15q21 and 15q22. Deletion at 15q11.2 (Prader-Willi/Angelman critical region) was excluded by FISH. To establish the exact breakpoints molecular studies were performed using bacterial artificial chromosome (BAC) clones spanning the 15q21.3 region. The absence of signal in this region defines the proband’s final karyotype as: 46,XX,del(15)(q21.3q21.3).ish del(15)(q21.3q21.3)(bA74K1-) Discussion: The authors emphasize the importance of complementary FISH and molecular studies in chromosomal abnormalities and compare the proband’s phenotype with similar cases described in the literature

Diagnóstico Citogenético em Líquidos Amnióticos Realizado entre 2000-2011 no Centro de Genética Médica Jacinto Magalhães, INSA, IP

Introdução: O diagnóstico pré-natal citogenético efetuado em líquido amniótico é um método seguro e fiável para deteção de anomalias cromossómicas fetais, sendo habitualmente realizado a partir das 15 semanas de gestação. Obtêm-se resultados, em média, após 8-10 dias de cultura dos amniócitos. Objectivo: Apresentar a estatística dos resultados obtidos na análise citogenética de líquidos amnióticos realizada na Unidade de Citogenética do Centro de Genética Médica Jacinto Magalhães entre 2000 e 2011, comparando-os com o descrito na literatura. Material e métodos: Entre janeiro de 2000 e dezembro de 2011 foram processados 10149 líquidos amnióticos. Os motivos para a realização da amniocentese foram, nomeadamente, idade materna avançada, anomalias ecográficas, marcadores ecográficos, rastreio bioquímico positivo, familiares com anomalias cromossómicas e risco de doença monogénica. Foram realizadas culturas de amniócitos de acordo com as técnicas convencionais de citogenética e os cromossomas identificados com bandas GTG ou GTL. Sempre que necessário efetuaram-se estudos de citogenética molecular (FISH) com as sondas adequadas ao esclarecimento do caso. Resultados: A análise revelou 342 cariótipos anormais (3,4%) dos quais 234 tinham anomalias numéricas e 108 estruturais. Os Síndromes de Down, de Edwards e de Turner foram as anomalias mais frequentes. Vinte e três culturas não cresceram, representando uma percentagem de 0,2% de insucesso. Conclusões: Os autores correlacionam os resultados obtidos com as indicações clínicas fornecidas e comparam-nas com o descrito na literatura. O presente estudo poderá ser utilizado para o estabelecimento de uma base de dados a nível nacional

Group and call effect in achieving success in a subject: Analyses of the "Biology and Botany" whole life in the E.U.I.T.A. (University of Seville)

La Biología y la Botánica son dos materias fundamentales en la formación de un ingeniero técnico agrícola. Sus contenidos se han ofrecido a veces en asignaturas independientes y, en otras ocasiones como la que nos ocupa, en una sola asignatura. Por otra parte, siempre se ha mirado con interés, e incluso preocupación, el efecto de la variable grupo, de forma independiente o relacionándola con su distribución en turnos de mañana o tarde, así como la importancia de la convocatoria (en los distintos momentos a lo largo del curso académico) en la que el alumno consigue superar la asignatura. Aquí presentamos los resultados en la consecución de objetivos por parte de los alumnos y, por ende, de los profesores y de la Universidad de Sevilla a lo largo de la totalidad de la vida de la asignatura "Biología y Botánica" dentro del plan de estudios 2003/04 para tres titulaciones simultáneas, desde su comienzo en el curso 2003/04 hasta su extinción en el curso 2009/10, de la E.U.I.T.A. (Escuela Universitaria de Ingeniería Técnica Agrícola) de la Universidad de Sevilla. Además se consideran convocatorias pertenecientes al período de extinción añadido. Los análisis toman en cuenta además el éxito en la superación de la asignatura tanto de forma cualitativa (aprobado o suspenso de la evaluación a la que el alumno se ha sometido) como cuantitativa (nota conseguida). Las conclusiones obtenidas permiten mirar los resultados y la consecución de objetivos, así como una potencial toma de decisiones para el futuro, basándose en un marco temporal amplio y objetivo.Biology and Botany are two critical issues in the formation of a technical agricultural engineer. Their topics are sometimes offered in separate subjects and, at other times as here, within a single one. Besides, it has always been considered with interest, and even concern, the effect of the variable group, independently or in relation to their distribution in the morning or afternoon turns, and the importance of the call (at different times during the academic year) at which the student gets to pass the subject. Here we present the results in the achievement of objectives by students, and therefore teachers and the University of Seville, along the entire life of the subject "Biology and Botany" within the teaching 2003/04 plan for three simultaneous academic programs, from its start in the course 2003/04 to its extinction in the course 2009/10, in the E.U.I.T.A. (University School of Technical Agricultural Engineering) of the University of Seville. Calls belonging to the period of extinction are also considered. Analyses are both qualitative, based on having success in passing the subject or not, and quantitative (mark scored). Obtained conclusions let us see the results and the achievement of objectives, as well as a potential decision-making for the future, with a base on a comprehensive and objective frame