Detection and quantification of selected cannabinoids in oral fluid samples by protein precipitation and LC-MS/MS.

Cannabis is the most widely consumed illicit drug worldwide. As consumption rates increase, partially due to the decriminalization of its use for medicinal and recreational purposes, analytical methods for monitoring different cannabinoids in several biological matrices have been developed. Herein, a simple and fast extraction procedure to extract natural cannabinoids from oral fluid (OF) samples was developed and fully validated according to the ANSI/ASB 2019 Standard Practices for Method Validation in Forensic Toxicology. Using only 0.2 mL of neat OF, the analytes [Δ9-tetrahidrocannabinol (THC), 11-hydroxy-Δ9-tetrahydrocannabinol (THC-OH), 11-nor-9-carboxy-Δ9-tetrahydrocannabinol (THC-COOH), cannabinol (CBN) and cannabidiol (CBD)] were extracted by protein precipitation with a mixture of methanol:acetonitrile (80:20, v/v); the extracts were centrifuged, evaporated to dryness and reconstituted in 100 µL of methanol. Analysis was performed by liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS). The developed methodology produced linear results for all compounds, with working ranges of 0.1-50 ng/mL for THC, 0.5-50 ng/mL for THC-OH, CBN and CBD, and 0.05-1 ng/mL for THC-COOH. Ion suppression was observed for THC, CBN and CBD, which did not impair sensitivity considering the low limits of quantification (LOQs) and limits of detection (LODs) obtained (which varied between 0.05 and 0.5 ng/mL). The extraction procedure produced great recoveries, and the compounds were stable. No interferences were found, and the method proved to be extremely fast, selective, precise, and accurate for use in routine analysis. The method was successfully applied to authentic samples.

Development and validation of a multi-substance method for routine analysis of pesticides in post-mortem samples by Ultra-High Performance Liquid Chromatography-Tandem Mass Spectrometry.

Pesticides play an important role in forensic toxicology and are usually classified as a single class of chemicals. Despite their commonly perceived unity, pesticides encompass a spectrum of compounds, including organophosphates, carbamates, pyrethroids or organochlorines, among others, each with varying degrees of toxicity. Pesticide analysis in post-mortem samples can be difficult due to the complexity of the samples and to the high toxicity of these compounds. The aim of this study was to develop and validate an easy to use, sensitive, and robust method, using ultra-performance liquid chromatography-tandem mass spectrometry to be incorporated in the routine flow for pesticide analysis in post-mortem blood samples. Described herein is a streamlined, expeditious, yet highly efficient method facilitating the screening, qualitative assessment, and quantitative confirmation of 15 pesticides, including acetamiprid, azinphos-ethyl, bendiocarb, carbofuran, chlorfenvinphos, dimethoate, imidaclopride, malathion, methiocarb, methomyl, parathion, pirimicarb, strychnine, tetrachlorvinphos, and thiacloprid in post-mortem blood, recognizing the pivotal role blood plays in forensic investigations. The developed method was linear from 10 to 200 ng/mL; limits of detection were between 1 and 10 ng/mL, depending on the compound; it was successfully evaluated a dilution ratio of 1-2, 5 and 10; and 8 substances showed maximum stability for the time interval studied. This UHPLC-MS/MS method is useful and a powerful tool in a toxicology lab because it is fast, simple, effective, and trustworthy. The results of this validation highlight the robustness of the analytical method, providing a valuable tool for the accurate and sensitive detection of pesticides in post-mortem blood. Poised for routine implementation, this method has already found success in suspected intoxication cases, promising to elevate the standards of forensic pesticide analysis.

Toxicological analysis of cocaine adulterants in blood samples.

Cocaine was the second most widely used drug in Europe in 2016, with 3.5 million consumers aged 15-64 years old. Adulterants are pharmacologically active substances developed for medical purposes, however, there is little knowledge about their influence in the human body when there is concomitant use with cocaine. The objective of this work was to validate a method that allows the identification, confirmation and quantification of cocaine adulterants in blood samples collected in vivo or post-mortem. The studied substances were atropine, phenacetin, hydroxyzine, ketamine, lidocaine and tetramisole. A retrospective study of the prevalence of these substances, as well as their relative concentrations, was made analysing 97 real blood samples previously tested positive for cocaine and/or its metabolites. The analytes of interest were extracted, using a simple method based on protein precipitation with frozen acetonitrile and further analysis by GC/MS. The method was fully validated in accordance with parameters and criteria implemented in the lab and SWGTOX recommendations (mean recovery: 94-115%; CV: 6.2-13%; BIAS: 2.7-7.8%). 31 samples were positive for adulterants: phenacetin (19%), tetramisole (15%), lidocaine (8%) and hydroxyzine (1%). Concentrations were higher in post-mortem samples for all compounds analysed. Lidocaine was more prevalent in samples collected in vivo whereas tetramisole was present almost exclusively in post-mortem samples. Phenacetin was evenly distributed between post-mortem and in vivo samples. The validated method allows rapid, precise, accurate and economic analysis of selected compounds and requires smaller sample aliquots which can be important in post-mortem cases. The information collected can be important in future studies of correlation between the presence of adulterants and cocaine toxicity.

Improved Sprouting and Growth of Mung Plants in Chromate Contaminated Soils Treated with Marine Strains of Staphylococcus Species.

Marine bacteria possess a wide variety of bioremediation potential which is beneficial environmentally and economically. In this study, bacterial isolates from marine waters were screened for tolerance and growth in high concentrations of chromate (Cr6+). Two isolates, capable of tolerating Cr6+ concentrations 300 µg mL-1 or higher, and found to completely reduce 20 µg mL-1 Cr6+ were grown in Cr6+ (50 and 100 mg kg-1) spiked garden soil. Notably, both facilitated normal germination and growth of mung (Vigna radiata) seeds, which could hardly germinate in Cr6+ spiked garden soil without either of these bacteria. In fact, large percent of mung seeds failed to sprout in the Cr6+ spiked garden soil and could not grow any further. Apparently, chromate detoxification by marine bacterial isolates and the ability of mung plants to deal with the reduced form appear to work complementarily. This study provides an insight into marine bacterial abilities with respect to chromium and potential applications in promoting growth of leguminous plants-similar to mung in particular-in Cr6+ contaminated soil.

Sobre queixas de dificuldades de memória na velhice / About complaints of difficulties memory in the aging

A queixa de "perda" ou "dificuldades de memória" entre idosos é ponto de discussão na literatura médica, especialmente na definição dos critérios para diagnóstico precoce da demência. Este trabalho problematiza a noção de dificuldades de memória na velhice. A partir da reflexão do filósofo Canguilhem, assume-se que reduzir o envelhecimento a aspectos unicamente biológicos significa assumir uma tendência reducionista. Nesta visada teórica, incluímos os sujeitos ­ suas queixas ­ numa discussão que envolve a dicotomia normal versus patológico quando está em causa a "perda" de memória em idosos. A procura de idosos por atividades que possam auxiliá-los a enfrentar problemas relativos à memória e a oferta de oficinas com tal finalidade tem-se constituído como uma tendência atual em serviços (públicos e privados) voltados para a promoção da saúde na terceira idade. Um relato de experiência com "oficinas de linguagem e memória" para idosos é apresentado nesta comunicação, bem como a discussão sobre essa experiência. (AU)

The complaint of "loss" or "memory difficulties" in the elderly is a point of discussion in the medical literature, especially in the definition of criteria for early diagnosis of dementia. This paper discusses the difficulties of memory in old aging. From the reflection of the philosopher Canguilhem, this communication assumed that reduce the aging aspects only biological reductionist means taking a position. In this theoretical offered, we included subjects - their complaints - the discussion that involves the usual dichotomy pathological X normal, when it is in question "loss" of memory in the elderly. The demand for older for activities that can help them address problems with memory and offer workshops for this purpose is a current tendency in services (public and private) to promotion of health in old age. This communication presented in the report of experience with "language and memory workshops" for the elderly. (AU)

Sequencing CYP2D6 for the detection of poor-metabolizers in post-mortem blood samples with tramadol.

Tramadol concentrations and analgesic effect are dependent on the CYP2D6 enzymatic activity. It is well known that some genetic polymorphisms are responsible for the variability in the expression of this enzyme and in the individual drug response. The detection of allelic variants described as non-functional can be useful to explain some circumstances of death in the study of post-mortem cases with tramadol. A Sanger sequencing methodology was developed for the detection of genetic variants that cause absent or reduced CYP2D6 activity, such as *3, *4, *6, *8, *10 and *12 alleles. This methodology, as well as the GC/MS method for the detection and quantification of tramadol and its main metabolites in blood samples was fully validated in accordance with international guidelines. Both methodologies were successfully applied to 100 post-mortem blood samples and the relation between toxicological and genetic results evaluated. Tramadol metabolism, expressed as its metabolites concentration ratio (N-desmethyltramadol/O-desmethyltramadol), has been shown to be correlated with the poor-metabolizer phenotype based on genetic characterization. It was also demonstrated the importance of enzyme inhibitors identification in toxicological analysis. According to our knowledge, this is the first study where a CYP2D6 sequencing methodology is validated and applied to post-mortem samples, in Portugal. The developed methodology allows the data collection of post-mortem cases, which is of primordial importance to enhance the application of these genetic tools to forensic toxicology and pathology.

Sexuality and HIV/AIDS in third age / Sexualidade e HIV/AIDS na terceira idade

This paper is specifically targeted at bringing up a reflection involving the articulation between the elderly and sexuality. Such theme has been motivated as from the verification, based on epidemiological data, that Brazil currently is facing an increasing number of new cases of AIDS among the over 60 age group. If, on the one hand, said data serves to cast off beliefs (stigmatized ones) toward the idea that "the elderly do not have sexual intercourses", on the other hand, it shows the need to question the reasons for such state of things. Therefore, attention has been paid to approaching "sexuality", "Third age" and “HIV/AIDS” as concepts and then explore how their association allows reading the reality mentioned above. At the end, there is a discussion about how, in Brazil, the State and non-governmental organizations have been facing the elderly health problem so as to see, more specifically, the social protection system available to face AIDS in this segment of population.

Este trabalho tem como objetivo encaminhar uma reflexão que envolve a articulação entre velhice e sexualidade. Esse tema foi motivado pela constatação, através de dados epidemiológicos, de que o Brasil assiste, atualmente, a um crescente aumento de novos casos de AIDS no grupo etário com idade superior a 60 anos. Se, de um lado, esse dado serve para desconstruir o imaginário (estigmatizado) de que “velho não faz sexo”, de outro, ele indica que é preciso problematizar as razões que respondem por esse estado de coisas. Por isso, procurou-se abordar “sexualidade”, “terceira idade” e “HIV/AIDS” como conceitos para, então, explorar de que modo sua associação permite ler a realidade antes referida. Ao final, há uma discussão sobre como, no Brasil, o Estado e as organizações não governamentais estão enfrentando o problema da saúde do idoso para, mais especificamente, deixar ver o sistema de proteção social disponível para o enfrentamento da AIDS neste segmento populacional.

Bioactivity-based screening methods for antibiotics residues: a comparative study of commercial and in-house developed kits.

Two bioactivity-based screening methods for antibiotic residue analysis (FAST Antimicrobial Screening Test and PremiTest) were compared, in terms of sensitivity, with a new in-house developed tube test assay using Escherichia coli. Tests were performed using antibiotic standards, spiked samples and real incurred samples. The minimum inhibitory concentration (MIC) for several antibiotics was established and compared with maximum residue levels (MRLs) in samples. The results of all evaluated tests are compared with liquid chromatography-tandem mass spectrometry multi-residue screening tests to compare parameters such as sample preparation, cost, time of analysis and confidence in results. For all tests, values of half the maximum residue limit (0.5 × MRL) were considered as a satisfactory target for a screening method. The potential and limitations of each method are discussed to indicate more rational and effective strategies for high-throughput residue monitoring and surveillance programmes. It was concluded that bioactivity-based screening methods are a useful tool, but the best compromise between minimum performance limits, cost and selectivity must be taken into account. For laboratories equipped with mass spectrometry, multi-class screening methods provide more specific responses with high sensitivity.

Sexualidade e AIDS na Terceira Idade / Sexuality and AIDS in the Third Age

Este trabalho tem como objetivo encaminhar uma reflexão que envolve a articulação entre velhice e sexualidade. Esse tema foi motivado pela constatação, através de dados epidemiológicos, de que o Brasil assiste, atualmente, a um crescente aumento de novos casos de AIDS no grupo etário com idade superior a 60 anos. Se, de um lado, esse dado serve para desconstruir o imaginário (estigmatizado) de que “velho não faz sexo”, de outro, ele indica que é preciso problematizar as razões que respondem por esse estado de coisas. Por isso, procurou-se abordar “sexualidade”, “terceira idade” e “HIV/AIDS” como conceitos para, então, explorar de que modo sua associação permite ler a realidade antes referida. Ao final, há uma discussão sobre como, no Brasil, o Estado e as organizações não governamentais estão enfrentando o problema da saúde do idoso para, mais especificamente, deixar ver o sistema de proteção social disponível para o enfrentamento da AIDS neste segmento populacional...

This paper aims to send a reflection that involves the relationship between age and sexuality. This theme was motivated by the realization, through epidemiological data, that Brazil assists, currently, an increasing number of new AIDS cases in the This theme was motivated by the realization, through epidemiological data, that Brazil assists, currently, an increasing number of new AIDS cases in the age group older than 60 years. If, on the one hand, this data serves to deconstruct the imagery (stigmatized) that "old does not have sex" on the other, it indicates that we must question the reasons that account for this state of affairs. Therefore, we sought to address "sexuality", "third age" and "HIV / AIDS" as concepts to then explore how their association allows you to read the reality referred to above. Finally, there is a discussion about how, in Brazil, the state and non-governmental organizations are facing the problem of health care for the elderly, more specifically, let see the social protection system available for coping with AIDS in this population segment...

Graduação em gerontologia na PUC-SP: o desafio da longevidade / Undergraduate course in gerontology at PUC-SP: the challenge of logenvity

A questão da longevidade tem-se imposto na pós-modernidade não só porque se vive mais como também mais tempo na velhice. A Gerontologia é um campo que transforma tal constatação numa proposição problemática, investigando o impacto social e subjetivo dessa “novidade”. Nessa perspectiva, o curso de graduação em Gerontologia da PUC-SP foi elaborado com base no compromisso de introduzir o aluno numa reflexão que parte do reconhecimento de que tanto o processo de envelhecimento quanto a velhice implicam complexidade. A imbricação interdisciplinar e integrativa ganha ênfase nesta proposta curricular e sustenta as atividades práticas de formação dos alunos.

The longevity issue has an important role in post-modernity, not only because people live more, but also because a long part of this life happens in old age. Gerontology is a field that turns this fact into a problematic proposition, investigating the social and subjective impact of this “news”. In this perspective, the Undergraduate Course in Gerontology of PUC-SP arose from the commitment to introducing the student to a reflection process based on the recognition that both aging and old age imply complexity. The interdisciplinary and integrative imbrication receives special emphasis in this curricular proposal and sustains the practical activities of students’ education.

Efeitos da escrita na clínica de linguagem / Efectos de la escritura en la clínica de lenguaje / Writing as a clinical device in speech clinic

Este artigo discute os efeitos clínicos da inclusão de uma atividade sistemática com a escrita no tratamento de crianças e de adultos cujas falas/escutas são sintomáticas. Questões relativas à natureza específica de atos que fundamentam a clínica de linguagem e seus desdobramentos teóricos estão no foco da reflexão encaminhada à luz de segmentos de sessões clínicas.

This paper aims at discussing some effects of writing as a therapeutic procedure in the so called Language Clinic. It is argued that its inclusion in the clinical scenario promotes changes not only in writing itself, but also in speech because it affects the listeners' listening to language. The specific nature of such an intervention is focused and the discussion carried on in this paper should provide specific and original theoretical support for clinicians and researchers in the field of speech therapy. Segments of clinical sessions will be presented and dealt with.

Este artículo se propone discutir los efectos de un trabajo sistemático con la escritura en el atendimiento clínico de niños e adultos que presentan síntomas en el lenguaje. Cuestiones relativas a la naturaleza específica de las acciones que fundamentan la clínica del lenguaje y sus desdoblamientos están en el foco de la reflexión encaminada. Análisis de segmentos de los atendimientos clínicos movimientam el debate.

A afasia e o problema da convergência entre teoria e abordagens clínicas / The aphasia and the problem of the convergence between theory and therapy

Objetiva tomar a afasia como lugar de discussão do problema teoria/ideologia no campo da patologia e clínica de linguagem