Really the best of both? Application of an mRNA/miRNA multiplex assay to casework samples, animal samples, and a storage study.

The identification of body fluids is an important area of forensic genetics. In particular, the susceptibility to degradation of casework samples is of crucial importance, as the traces can often be exposed to different environmental conditions over a long period of time. RNAs especially are used as molecular markers for the identification of body fluids in forensics. Messenger RNAs (mRNAs) show an increased susceptibility to degradation, e.g. under humidity and UV radiation but are highly body fluid-specific. The shorter micro RNAs (miRNAs), however, are less susceptible to degradation, but only a few body fluid-specific markers could be investigated. In this study, a self-developed mRNA/miRNA multiplex assay for capillary electrophoresis from a preliminary study was further adapted and validated. The approach was applied to casework samples, animal samples, and a storage study. The advantages and disadvantages of the mRNA/miRNA assay were investigated in order to review a possible application for forensic casework. Some miRNA markers were also detected in animal samples, which once again underlines the possible non-specificity of miRNAs. In the storage study, the different markers were detected for different lengths of time depending on the body fluid examined. For almost all body fluids, the miRNA markers were still detectable after a period of 35 days under environmental conditions compared to the mRNA markers. The mRNA peaks were often already clearly reduced or no longer detectable after 14 days. The results show the advantage of the new mRNA/miRNA assay compared to established mRNA approaches, especially for older and degraded samples, but the assay has its limitations due to the limited number of specific miRNA markers.

Best of both: A simultaneous analysis of mRNA and miRNA markers for body fluid identification.

The body fluid identification of traces found at crime scenes is crucial in relation of the circumstance of crime. For this reason, the body fluid identification (BFI) by molecular biological methods has been increasingly investigated in recent decades. Especially the use of messenger RNA (mRNA) has been established and validated by various studies. mRNAs can resist degradation for several decades under dry and dark environmental conditions, but degradation increases greatly e.g., in humid environments and UV radiation. In contrast, the shorter and protein-protected micro RNAs (miRNAs) are less susceptible to degradation, but not all potential markers are tissue-specific. The aim of this study was to develop a simultaneous mRNA/miRNA multiplex assay to take advantage of both types of RNA. The final assay was tested for various body fluids, dilutions, and mixtures. To demonstrate the advantage of a combined mRNA/miRNA assay, older and mostly degraded samples were examined and compared to an established mRNA assay. Initial results from degraded samples show that tissue-specific miRNAs expected could be detected for 93% of the degraded samples compared to mRNA markers with 25% of the mRNA assay. The result is a simultaneous mRNA/miRNA multiplex assay on capillary electrophoresis (CE) for the first time.

A constituição do sujeito na psicanálise lacaniana: impasses na separação / The constitution of the subject in lacanian psychoanalysis: impairments in separation / La constitución del sujeto en el psicoanálisis lacaniano: impases en la separación

Este trabalho consiste em uma pesquisa teórica a respeito da constituição do sujeito no âmbito da psicanálise lacaniana. A vertente clínica que motiva esta pesquisa é o atendimento de crianças com problemas graves (psicose, autismo, deficiência mental, etc.) e suas mães, e das dificuldades encontradas num momento privilegiado de seu tratamento, em que ocorreria a separação estrutural entre ambas. Há dois modos pelos quais se constitui o sujeito: segundo o Estádio do Espelho e segundo a topologia da alienação e separação. Ambos são apresentados. Ao explicar a alienação, o estudo enfoca o surgimento do sujeito no inconsciente, distinto do eu, que é essencialmente imaginário. Este surgimento é apresentado de acordo com a leitura lacaniana do cogito ergo sum de Descartes. Os impasses encontrados na clínica quando da separação - especialmente o surgimento de sintomas físicos na mãe - são considerados e analisados à luz das teorias vigentes.

A theoretical research on the constitution of the subject in Lacanian Psychoanalysis is provided. The clinical aspect which motivates current research is the dealing with children bearing serious problems (psychosis, autism, mental disability, etc) and with their mothers, coupled to difficulties encountered at a specific moment in the treatment, or rather, when the structural separation between mother and child occurs. There are two ways through which the subject constitutes itself: the mirror phase and the topology of alienation and separation are both carefully presented and investigated. Although alienation is analyzed, the study focuses on the emergence of the subject within the unconscious, as distinct from the ego which is essentially imaginary. Emergence is given according to the Lacanian reading of Descartes's Cogito, ergo sum. Impairments in the clinical treatment when separation occurs - especially the emergence of physical symptoms in the mother - are taken into account and analyzed in the light of the above-mentioned theories.

Este trabajo consiste en una investigación teórica respecto a la constitución del sujeto en el ámbito del psicoanálisis lacaniano. La vertiente clínica que motiva esta investigación es la atención a niños con problemas graves (psicosis, autismo, deficiencia mental, etc.) y sus madres, y las dificultades encontradas en un momento privilegiado de su tratamiento, en el que ocurriría la separación estructural entre ambas. Hay dos modos por los que se constituye el sujeto: según el Estadio del Espejo y según la topología de la alienación y separación. Ambos son presentados. Al explicar la alienación, el estudio enfoca el surgimiento del sujeto en el inconsciente, distinto del yo, que es esencialmente imaginario. Este surgimiento está presentado de acuerdo con la lectura lacaniana del cogito ergo sum de Descartes. Los impases encontrados en la clínica cuando de la separación - especialmente el surgimiento de síntomas físicos en la madre - son considerados y analizados a la luz de las teorías vigentes.

A alienaçäo eternizada: uma abordagem estrutural de um caso clínico / The eternalized alienation: a structural approach of a clinical case

Este trabalho é uma aplicaçäo de uma estratégia clínica baseada no conceito de alienaçäo, tal como definido por Jacques Lacan, para o atendimento de uma criança considerada psicótica e sua mäe. Tal estratégia, proposta por Jussara Falek Brauer, prescreve um atendimento individual, com uma sessäo semanal, para a criança e para a mäe. Considera a estrutura familiar e pretende lidar com as determinaçöes estruturais sobre a constituiçäo do Sujeito. Dois momentos säo especialmente focados - quando o trabalho é iniciado e outro posterior que exemplificam a mudança de posicionamento dessa criança, em funçäo do processo terapêutico em curso. Os conceitos teóricos que embasam esta abordagem - a Alienaçäo, a Topologia Lacaniana da Constituiçäo do Sujeito, o Significante, o Inconsciente, a Castraçäo, o Fetiche, a Perversäo, a Separaçäo - säo apresentados e relacionados ao material clínico. Algumas hipóteses säo apresentadas a respeito da estrutura psíquica da mäe, considerando a evoluçäo observada e as mudanças de posicionamento dessas pessoas, que levam ao estabelecimento de uma hipótese diagnóstica para a filha