Comparison of endogenous GHB concentrations in blood and hair in death cases with emphasis on the post mortem interval.

Gamma-hydroxybutyric acid (GHB) is an endogenous compound which has a story of clinical use and illicit abuse since the 1960's. The possibility to use a multi-sample approach for GHB evaluation, including whole blood and hair, to better characterize a forensic toxicology case and evaluate a possible causal association with the death is an exciting up-to-date issue. In addition, its post-mortem behaviour, namely regarding degradation and metabolism, has been increasingly investigated as a putative biomarker for post-mortem interval (PMI) estimation. Thus, in order to contribute to clarification of this specific aspect, whole blood and hair post-mortem GHB levels were evaluated in 32 real cases with previous information on death and autopsy data. The results obtained suggest that the PMI (until 5 days between death and sampling) influences GHB whole blood concentration, but not GHB levels in hair samples. No differences were encountered for the other parameters evaluated, including age, gender, cause of death and presence or absence of substances. This study brings new insights regarding the usefulness of GHB levels in forensic toxicology, which might be further strengthened with larger, but comparable, studies from other laboratories and institutions in the context of legal medicine.

Spatial and temporal assessment of crack cocaine use in 13 European cities through wastewater-based epidemiology.

Already in early 2000s, concerns have been growing in the EU about increasing use of cocaine and it is estimated that below 1 % of the population administer the drug by smoking crack cocaine. New available data suggests an increase in the use of crack cocaine and an increase in the number of crack cocaine users entering treatment has been reported in several European countries. Robust estimations of crack cocaine use are however not available yet. The use of crack cocaine has long been associated with severe adverse socio-economic conditions as well as mental health problems, such as suicide ideation and depression. The aim of this study was to assess spatial trends in population-normalized mass loads of crack cocaine biomarkers (i.e., anhydroecgonine and anhydroecgonine methyl ester) in 13 European cities in six countries (the Netherlands, Belgium, Ireland, Portugal, Spain and Italy). Furthermore, temporal trends over a five-year period were evaluated through the analysis of historic samples collected in the Netherlands. Finally, the stability of the crack cocaine biomarkers in wastewater was investigated through batch experiments. The samples were analyzed with a new developed and validated hydrophilic interaction liquid chromatography coupled to mass spectrometry method. Targeted crack cocaine biomarkers were found in all cities. Also, crack cocaine biomarker was detected in wastewater from 2017 to 2021 in the Netherlands, but no significance between the years were found. With respect to biomarker in-sample stability, AEME was found to be stable in wastewater. This study assessed crack cocaine use for the first time on a broad scale, both temporal and in cities across Europe, with wastewater-based epidemiology and it shows the importance of wastewater analysis to monitor community loads of crack cocaine use.

Qualitative and quantitative analysis of THC, 11-hydroxy-THC and 11-nor-9-carboxy-THC in whole blood by ultra-performance liquid chromatography/tandem mass spectrometry.

A qualitative and quantitative analytical method was developed for the simultaneous determination of Δ(9) -tetrahydrocannabinol (THC), 11-hydroxy-Δ(9) -tetrahydrocannabinol (11-OH-THC) and l1-nor-9-carboxy-Δ(9) -tetrahydrocannabinol (THC-COOH) in whole blood. The samples were prepared by solid-phase extraction followed by ultra-performance liquid chromatography/tandem mass spectrometry (UPLC/MS/MS) analysis using positive ion electrospray ionization and multiple reaction monitoring. The chromatographic separation was performed with an Acquity UPLC® HSS T3 (50 × 2.1 mm i.d., 1.8 µm) reversed-phase column using a methanol/2 mM ammonium formate (formic acid 0.1%) gradient in a total run time of 9.5 min. MS/MS detection was achieved with two precursor-product ion transitions per substance. The method was fully validated, including selectivity and capacity of identification, according to the identification criteria (two transitions per substance, signal-to-noise ratio, relative retention time and ion ratio) without the presence of interferences, limit of detection (0.2 µg/L for THC and 0.5 µg/L for 11-OH-THC and THC-COOH), limit of quantitation (0.5 µg/L for all cannabinoids), recovery (53-115%), carryover, matrix effect (34-43%), linearity (0.5-100 µg/L), intra-assay precision (CV < 10% for the relative peak area ratios and <0.1% for the relative retention time), inter-assay accuracy (mean relative error <10%) and precision (CV <11%). The method has already been successfully used in proficiency tests and subsequently applied to authentic samples in routine forensic analysis.

Trends in MDMA-related mortality across four countries.

AIMS: To determine trends in 3,4 methylenedioxymethamphetamine (MDMA)-related death rates across Australia, Finland, Portugal and Turkey and to analyse the toxicology and causes of death across countries. DESIGN: Analysis of MDMA-related deaths extracted from a national coronial database in Australia (2001-19) and national forensic toxicology databases in Finland (2001-17), Portugal (2008-19) and Turkey (2007-17). Presentation of MDMA use and seizure data (market indicators). SETTING: Australia, Finland, Portugal and Turkey. CASES: All deaths in which MDMA was considered by the forensic pathologist to be contributory to death. MEASUREMENTS: Information collected on cause and circumstances of death, demographics and toxicology. FINDINGS: A total of 1400 MDMA-related deaths were identified in Turkey, 507 in Australia, 100 in Finland and 45 in Portugal. The median age ranged from 24 to 27.5 years, and males represented between 81 and 94% of the deaths across countries. Standardized mortality rates significantly increased across all four countries from 2011 to 2017 during a period of increased purity and availability of MDMA. The underlying cause of death was predominantly due to drug toxicity in Australia (n = 309, 61%), Finland (n = 70, 70%) and Turkey (n = 840, 60%) and other causes in Portugal (n = 25, 56%). Minorities of all deaths across the countries were due to MDMA toxicity alone (13-25%). These deaths had a significantly higher blood MDMA concentration than multiple drug toxicity deaths in Australia, Finland and Turkey. Drugs other than MDMA commonly detected were stimulants (including cocaine, amphetamine and methamphetamine) (Australia 52% and Finland 61%) and alcohol (Australia 46% and Portugal 49%). In addition to MDMA toxicity, benzodiazepines (81%) and opioids (64%) were commonly identified in these deaths in Finland. In comparison, synthetic cannabinoids (15%) and cannabis (33%) were present in a minority of deaths in Turkey. CONCLUSIONS: Deaths related to 3,4 methylenedioxymethamphetamine (MDMA) increased in Australia, Finland, Portugal and Turkey between 2011 and 2017. Findings show MDMA toxicity alone can be fatal, but multiple drug toxicity remains more prevalent.

Spatio-temporal assessment of illicit drug use at large scale: evidence from 7 years of international wastewater monitoring.

BACKGROUND AND AIMS: Wastewater-based epidemiology is an additional indicator of drug use that is gaining reliability to complement the current established panel of indicators. The aims of this study were to: (i) assess spatial and temporal trends of population-normalized mass loads of benzoylecgonine, amphetamine, methamphetamine and 3,4-methylenedioxymethamphetamine (MDMA) in raw wastewater over 7 years (2011-17); (ii) address overall drug use by estimating the average number of combined doses consumed per day in each city; and (iii) compare these with existing prevalence and seizure data. DESIGN: Analysis of daily raw wastewater composite samples collected over 1 week per year from 2011 to 2017. SETTING AND PARTICIPANTS: Catchment areas of 143 wastewater treatment plants in 120 cities in 37 countries. MEASUREMENTS: Parent substances (amphetamine, methamphetamine and MDMA) and the metabolites of cocaine (benzoylecgonine) and of Δ9 -tetrahydrocannabinol (11-nor-9-carboxy-Δ9 -tetrahydrocannabinol) were measured in wastewater using liquid chromatography-tandem mass spectrometry. Daily mass loads (mg/day) were normalized to catchment population (mg/1000 people/day) and converted to the number of combined doses consumed per day. Spatial differences were assessed world-wide, and temporal trends were discerned at European level by comparing 2011-13 drug loads versus 2014-17 loads. FINDINGS: Benzoylecgonine was the stimulant metabolite detected at higher loads in southern and western Europe, and amphetamine, MDMA and methamphetamine in East and North-Central Europe. In other continents, methamphetamine showed the highest levels in the United States and Australia and benzoylecgonine in South America. During the reporting period, benzoylecgonine loads increased in general across Europe, amphetamine and methamphetamine levels fluctuated and MDMA underwent an intermittent upsurge. CONCLUSIONS: The analysis of wastewater to quantify drug loads provides near real-time drug use estimates that globally correspond to prevalence and seizure data.

Liquid chromatography/tandem mass spectrometry for the qualitative and quantitative analysis of illicit drugs and medicines in preserved oral fluid.

A qualitative and quantitative analytical method was developed for the simultaneous determination of 24 illicit drugs and medicines, in preserved oral fluid samples collected with the StatSure Saliva Sampler collection device. The samples were prepared by liquid-liquid extraction followed by liquid chromatography/tandem mass spectrometry (LC/MS/MS) analysis. The chromatographic separation was performed with an Atlantis T3 (100 x 2.1 mm i.d., 3 microm) reversed-phase column using an acetonitrile/2 mM ammonium formate buffer pH 3.4 gradient and the MS/MS detection was achieved with two precursor-product ion transitions per substance. The method was fully validated, including specificity and capacity of identification, limit of detection (0.2-2.1 microg/L), limit of quantitation (0.8-6.4 microg/L), recovery (34-98%), carryover, linearity (the method was linear in the range 1-200 microg/L), intra-assay precision (coefficient of variance (CV) <20% for 20 microg/L and CV <10% for 100 microg/L) and inter-assay accuracy (mean relative error <15%) and precision (CV <20%). The method showed to be specific and sensitive. It has already been successfully used in four proficiency tests and subsequently applied to oral fluid samples collected from road traffic volunteers in the driving population of Portugal (districts of Lisbon, Coimbra and Porto), within the DRUID project.

Dried blood spots combined to an UPLC-MS/MS method for the simultaneous determination of drugs of abuse in forensic toxicology.

A method for the simultaneous determination of 11 illicit drugs, using the dried blood spot (DBS) sampling technique combined with the UPLC-MS/MS technology was developed to study its applicability within the forensic toxicology. The DBS samples, prepared from a blood volume of 50µL and using the Whatman® BFC 180 bloodstain cards, were extracted with a methanol/acetonitrile mixture. The chromatographic separation was performed using an Acquity UPLC® HSS T3 column (100mm×2.1mm, 1.8µm) and an acetonitrile/2mM ammonium formate (0.1% formic acid) gradient. The detection was accomplished with a TQ Detector, operating in the ESI+ and MRM modes. The method was validated in terms of selectivity, matrix effect, extraction recovery (42%-91%), carryover, LOD and LOQ (0.5-1ng/mL and 1-5ng/mL, respectively), linearity (LOQ to 500ng/mL), intraday and interday precision (3.8-14% and 5.3-13%, respectively), accuracy (-9.3% to 7.9%) and dilution integrity. An eight months stability study at room temperature, 2-8°C and -10°C, was also performed, with the best results obtained at -10°C. The procedure was applied to 64 real samples (92 positive results for substances included in this study). The results were compared with the methodologies routinely applied in the laboratory and the statistical analysis allowed to establish an acceptable correlation. This study permitted to determine that the DBS can represent an alternative or a complement to conventional analytical and sampling techniques, responding to some of the present issues concerning the different forensic toxicology applications.

Contactless decontamination of hair samples: cannabinoids.

Room temperature ionic liquids (ILs) have already been shown to provide efficient extraction media for several systems, and to capture volatile compounds, namely opiates. In this work, a novel, contactless, artefact-free extraction procedure for the removal of Δ9 -tetrahrydrocannabinol (THC) from the surface of human hair is presented. To prepare in vitro cannabinoids-contaminated hair, samples were flushed with hashish smoke for 7 h. The decontamination experiments were carried at 100 °C for 24 h, according to the procedure previously described. Fifty-three ILs were screened and presented decontamination efficiencies ranging from 0 to 96 %. Although the majority of the ILs presented efficiencies above 90%, the 1-ethanol-3-methyl tetrafluoroborate (96%) was chosen for further process optimization. The Design of Experiments results demonstrated that all studied variables were significant for the process and the obtained optimum conditions were: 100 °C, 13 h and 175 mg of IL. In the work of Perrotin-Brunel et al. (J. Mol. Struct. 2011, 987, 67), it is demonstrated that, at 100 °C, full conversion of tetrahydrocannabinolic acid (THCA) into THC is obtained after 60 min. Since our decontamination takes place over 13 h at 100 °C, full conversion of THCA into THC is expected. Additionally, our method was compared with the method proposed by Cairns et al. (Forensic Sci. Int. 2004, 145, 97), through the analysis of 15 in vitro contaminated hair samples. The results demonstrated that with our method a mean extraction efficiency of 11 % higher was obtained. Copyright © 2016 John Wiley & Sons, Ltd.

Assessment of Specificity of the Badcamp Agility test for Badminton Players.

The Badcamp agility test was created to evaluate agility of badminton players. The Badcamp is a valid and reliable test, however, a doubt about the need for the use of this test exists as simpler tests could provide similar information about agility in badminton players. Thus, the aim of this study was to examine the specificity of the Badcamp, comparing the performance of badminton players and athletes from other sports in the Badcamp and the shuttle run agility test (SRAT). Sixty-four young male and female athletes aged between 14 and 16 years participated in the study. They were divided into 4 groups of 16 according to their sport practices: badminton, tennis, team sport (basketball and volleyball), and track and field. We compared the groups in both tests, the Badcamp and SRAT. The results revealed that the group of badminton players was faster compared to all other groups in the Badcamp. However, in the SRAT there were no differences among groups composed of athletes from open skill sports (e.g., badminton, tennis, and team sports), and a considerable reduction of the difference between badminton players and track and field athletes. Thus, we concluded that the Badcamp test is a specific agility test for badminton players and should be considered in evaluating athletes of this sport modality.

A fast and reliable method for GHB quantitation in whole blood by GC-MS/MS (TQD) for forensic purposes.

Gamma-hydroxybutyric acid (GHB) is an endogenous compound with a story of clinical use since the 1960s. However, due to its secondary effects, it has become a controlled substance, entering the illicit market. A fully validated, sensitive and reproducible method for the quantification of GHB by methanolic precipitation and GC-MS/MS (TQD) in whole blood is presented. Using 100µL of whole blood, obtained results included a LOD and LLOQ of 0.1mg/L and a recovery of 86% in a working range between 0.1 and 100mg/L. This method is sensitive and specific to detect the presence of GHB in small amounts of whole blood (both ante-mortem or post-mortem), and is, to the authors' knowledge, the first GC-MS-MS TQD method that uses different precursor ions and product ions for the identification of GHB and GHB-D6 (internal standard). Hence, this method may be especially useful for the study of endogenous values in this biological sample.

Fundamentos del Diseño Universal para el Aprendizaje Desde la Perspectiva Internacional / Fundamentals of Universal Design for Learning from an International Perspective

RESUMEN: El presente estudio se enmarca en el ámbito educativo, concretamente, en el área de la educación inclusiva, profundizando en el Diseño Universal para el Aprendizaje (DUA). El DUA se concibe como un enfoque teórico y práctico que actúa como una herramienta eficaz para alcanzar una educación de calidad y equidad dentro del movimiento de la inclusión educativa. El objetivo de este estudio es analizar la presencia de los fundamentos del DUA (derecho de la educación; diversidad; Diseño Universal; currículo flexible; igualdad de oportunidades; neurociencia y tecnologías de la información y la comunicación) en los documentos publicados por los organismos internacionales con competencia en materia educativa y de repercusión mundial. La metodología de la investigación es de corte cualitativo utilizando el análisis documental. Las categorías de análisis corresponden a los siete fundamentos del DUA. Las fuentes documentales para el estudio han sido fundamentalmente las páginas web de los organismos internacionales y la base de datos UNESDOC, obteniéndose una muestra definitiva de once documentos pertenecientes a la literatura gris. Tras la revisión documental, los resultados han evidenciado que no todos los documentos objeto de estudio contemplan en su contenido las categorías de análisis. En suma, se observan carencias en cuanto a la presencia de los siete fundamentos del DUA en los documentos publicados analizados, por lo que se requiere de un nuevo tratamiento a nivel internacional del DUA con el fin de promover una educación de calidad e inclusiva, que atiende y respeta a las diferencias individuales dentro del proceso de enseñanza-aprendizaje.

ABSTRACT: The present study was carried out in the frame of the educational field, specifically in the area of​​inclusive education, going deeper into the Universal Design for Learning (UDL). UDL is conceived as a theoretical and practical approach that acts as an effective tool to achieve quality education and equity within the educational inclusion movement. The objective of this study is to analyze the presence of the fundamentals of UDL (education law; diversity; Universal Design; flexible curriculum; equal opportunities; neuroscience and information and communication technologies) in documents published by international organizations with competence in education and global impact. The research methodology is of a qualitative nature and uses documentary analysis. The categories of analysis correspond to the seven fundamentals of UDL. The documentary sources of the study were mainly the web pages of international organizations and the UNESDOC database, obtaining a definitive sample of eleven documents belonging to the gray literature. After the document review, the results showed that not all documents under study include the categories of analysis in their content. In summary, there are gaps in the presence of the seven fundamentals of UDL in the published documents analyzed, which is why a new international treatment of the UDL is needed to promote an inclusive and quality education that meets and respects individual differences within the teaching-learning process.

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.

Development, optimization, and validation of a novel extraction procedure for the removal of opiates from human hair's surface.

Room temperature ionic liquids (ILs) have proved to be efficient extraction media for several systems, and their ability to capture volatile compounds from the atmosphere is well established. We report herein a contactless extraction procedure for the removal of opiate drugs from the surface of human hair. The compounds were chosen as a model drug, particularly due to their low volatility. Equal amounts of IL and hair (about 100 mg) were introduced in a customized Y-shaped vial, and the process occurred simply by heating. After testing several ILs, some of them (e.g. 1-methyl-3-ethanol-imidazolium tetrafluoroborate, phenyl-trimethyl-ammonium triflate or bis(dimethyl) diheptylguanidinium iodide) showed extraction efficiencies higher than 80% for the two studied compounds, morphine and 6-monoacetylmorphine. Using the design of experiments (DOE) approach as an optimization tool, and bearing in mind the hygroscopic properties of the ILs (in particular, 1-methyl-3-ethanol-imidazolium tetrafluoroborate), the process was optimized concerning the following variables: temperature (50-120 ºC), extraction time (8-24 h), IL amount (50-200 mg) and water content of the IL (0.01-60%). This study not only provided the optimum conditions for the process (120 ºC, 16 h, 100 mg of IL containing 40% of water), but has also showed that the water content of the IL represents the variable with the most significant effect on the extraction efficiency. Finally, we validated our method through the comparison of the results obtained by treating hair samples with the described procedure to those obtained using a standard washing method and criteria for positivity.

Gamma-hydroxybutyric acid endogenous production and post-mortem behaviour - the importance of different biological matrices, cut-off reference values, sample collection and storage conditions.

Gamma-Hydroxybutyric Acid (GHB) is an endogenous compound with a story of clinical use, since the 1960's. However, due to its secondary effects, it has become a controlled substance, entering the illicit market for recreational and "dance club scene" use, muscle enhancement purposes and drug-facilitated sexual assaults. Its endogenous context can bring some difficulties when interpreting, in a forensic context, the analytical values achieved in biological samples. This manuscript reviewed several crucial aspects related to GHB forensic toxicology evaluation, such as its post-mortem behaviour in biological samples; endogenous production values, whether in in vivo and in post-mortem samples; sampling and storage conditions (including stability tests); and cut-off reference values evaluation for different biological samples, such as whole blood, plasma, serum, urine, saliva, bile, vitreous humour and hair. This revision highlights the need of specific sampling care, storage conditions, and cut-off reference values interpretation in different biological samples, essential for proper practical application in forensic toxicology.

Validation and application of an UPLC-MS/MS method for the quantification of synthetic cannabinoids in urine samples and analysis of seized materials from the Portuguese market.

An UPLC-MS/MS method using ESI+ionization and MRM was developed and fully validated according to international guidelines for the qualitative and quantitative analysis of nine synthetic cannabinoids and/or their metabolites in urine samples (1mL). Prior to extraction the samples were subjected to an enzymatic hydrolysis using ß-glucuronidase followed by a SPE procedure using Oasis(®) HLB 3cc (60mg) columns. The chromatographic separation was performed with an Acquity UPLC(®) HSS T3 (50mm×2.1mm i.d., 1.8µm) reversed-phase column using a gradient with methanol-ammonium formate 2mM (0.1% formic acid) and with a run time of 9.5min. The method was validated in terms of selectivity, capacity of identification, limits of detection (0.01-0.5ng/mL) and quantification (0.05-0.5ng/mL), recovery (58-105%), carryover, matrix effect, linearity (0.05-50ng/mL), intra-assay precision, inter-assay accuracy and precision (CV<20%). The method was applied to 80 authentic samples, five of them (6.2%) were confirmed or suspected to be positive for the metabolites JWH-018 N-hydroxypentyl and JWH-018 N-pentanoic acid of JWH-018 and for the metabolite JWH-122 N-(5-hydroxypentyl) of JWH-122, and three of them in association with THC and/or THCCOOH (substances included in the method, together with the 11-OH-THC). Additionally, 17 spice products were analyzed, for which were confirmed the presence of the following substances: AM-2201, JWH-018, JWH-022 JWH-073, JWH-122, JWH-203, JWH-210, JWH-250, HU-210 and RCS-4, according to the comparison with authentic reference material and published data. The analytical method developed allowed the analysis of synthetic cannabinoids and the notification of the first cases in Portugal.

Prevalence of alcohol, illicit drugs and psychoactive medicines in killed drivers in four European countries.

Our objective was to determine the presence of psychoactive substances in blood of drivers killed in road crashes in four European countries. Data from 1118 drivers of car and vans, killed between 2006 and 2009, were collected in Finland, Norway, Portugal and Sweden. The prevalence of any psychoactive substance ranged between 31 and 48%. Alcohol (≥ 0.1 g/L) was the most common finding, 87% had a blood alcohol concentration (BAC) ≥ .5 g/L. Benzodiazepines (1.8-13.3%) and amphetamines (0-7.4%) were the most prevalent psychoactive medicines and illicit drugs, respectively. Alcohol-drug and drug-drug combinations were rather prevalent. Differences in alcohol/drug findings seemed to reflect differences in use in the countries. More research should be done to develop preventive strategies to reduce the number of alcohol- and drug-related traffic accidents targeting at-risk groups, such as drivers with very high BACs and novice drivers.

Tratamento pediátrico de grande queimado agudo: Manejo clínico, cirúrgico e uso de matriz de regeneração dérmica / Pediatric treatment of large acute burn: Clinical, surgical management and use of dermal regeneration matrix / Tratamiento pediátrico de gran quemado agudo: Manejo clínico, quirúrgico y uso de matriz de regeneración dérmica

Objetivo: Analisar o tratamento de queimaduras em fase aguda, conduta clínica, cirúrgica e uso de matriz de regeneração dérmica (MRD) em criança internada na Unidade de Queimados do Hospital Regional da Asa Norte (UQ-HRAN), Brasília-DF. Relato do Caso: D.M.R., 2 anos e 7 meses, masculino, vítima de queimadura por chama direta, 60% de superfície corporal acometida, 50% sendo de espessura total. Atendido inicialmente segundo Protocolo de Rotinas da UQ-HRAN, entretanto, evoluiu com piora clínica, necessitando de Unidade de Terapia Intensiva pediátrica, por 58 dias. Neste período, foi submetido a 2 escarotomias, 3 desbridamentos e 10 hemotransfusões, apresentou infecções nas áreas queimadas e variados esquemas antibióticos. Tendo em vista a extensão de áreas acometidas, restrição de áreas doadoras viáveis e impossibilidade de outros curativos, optou-se pelo implante de MRD no 16° dia de internação e, após a integração, foram realizadas 6 enxertias cutâneas. Recebeu alta no 118° dia de internação. Conclusão: Houve necessidade de assistência clínica e cuidados intensivos, além de utilização de técnica cirúrgica com implante de MRD em grande queimado agudo. Sendo assim, a MRD teve como objetivo proporcionar leito receptor de qualidade, associado a maior integração para enxertia cutânea futura, pois a carência de áreas doadoras o colocaria em risco de vida maior. Esse conjunto de fatores contribuiu para o sucesso do tratamento e a boa recuperação da criança.

Objective: To analyze the treatment of burn in acute-phase using clinical, surgical management and use of dermal regeneration matrix (DRM) in a child hospitalized at the Burn Unit of Asa Norte Regional Hospital (BU-ANRH), Brasília-DF. Case Report: D.M.R., 2 years and 7 months old, male, victim of direct fire, 60% of body surface affected, 50% full-thickness. Initially treated according to the BU-ANRH Routine Protocol, however, evolved to clinical decline, requiring a pediatric Intensive Care Therapy for 58 days. During this period, he underwent two escharotomies, three debridements and 10 blood transfusions. He exhibited infection in burned areas and used multiple antibiotic schemes. Due to the extension of affected surface, restriction of viable donor zone and unusable of other bandages, DRM was implanted on the 16th day of hospitalization and integration, he has done six cutaneous grafting. He was discharged on the 118th day of hospitalization. Conclusion: Clinical and intensive care were needed, besides the use of surgical technique with DRM implantation in a large acute burn. Thus, DRM has had as a goal to provide quality receptor bed, associated with greater integration for future cutaneous grafting, since the lack of donor areas would put it at greater risk of life. This set of factors contributed to the success of the treatment and the good recovery of the child.

Objetivo: Analizar el tratamiento de quemaduras en fase aguda, conducta médica clínica, quirúrgica y uso de matriz de regeneración dérmica (MRD) en niño internado en la Unidad de Quemados del Hospital Regional de Asa Norte (UQ-HRAN), Brasília-DF. Reporte de Caso: D.M.R., 2 años y 7 meses, masculino, víctima de quemadura de llama directa, 60% de la superficie del cuerpo afectada, 50% del espesor total. Inicialmente tratado de acuerdo con el Protocolo de Rutina UQ-HRAN, sin embargo, evolucionó con un empeoramiento clínico, necesitando cuidados en la Unidad de Terapia Intensiva pediátrica durante 58 días. Durante este período, fue sometido a 2 escarotomías, 3 desbridamientos y 10 transfusiones de sangre. Presentó infecciones en zonas quemadas y variados esquemas antibióticos. Debido a la extensión de las áreas afectadas, la restricción de las áreas viables de los donantes y la imposibilidad de otros curativos, se optó por el implante de MRD en el 16° día de internación y después de la integración, se realizaron 6 injertos cutáneos. Recibió alta en el 118° día de internación. Conclusión: Hubo necesidad de asistencia clínica y cuidados intensivos, además de utilización de técnica quirúrgica con implante de MRD en gran quemado agudo. Por lo tanto, la MRD tuvo como objetivo proporcionar lecho receptor de calidad, asociado a la mayor integración para el injerto cutáneo futuro, pues la carencia de áreas donantes lo pondría en mayor riesgo de vida. Este conjunto de factores, contribuyó al éxito del tratamiento y la buena recuperación del niño.

Quality of life in adult intradural primary spinal tumors: 36-Item Short Form Health Survey correlation with McCormick and Aminoff-Logue scales.

OBJECT: The most appropriate method to determine the quality of life of patients with intradural primary spinal tumors (IPSTs) is not still well established. METHODS: Clinical data in 234 patients who underwent surgery for intradural spinal disease were collected prospectively. The 36-Item Short Form Health Survey (SF-36), a generic score scale, was administered to 148 patients with IPSTs to demonstrate if the survey can be used to effectively evaluate these patients. Forty-eight patients were excluded because they did not complete the protocol. The study was finally conducted with 100 patients (45 male and 55 female) with IPSTs, and the results were compared with those of 2 other scales: the McCormick scale and the Aminoff-Logue scale. RESULTS: Construct validity was demonstrated by confirming the hypothesized relationship between the scores of the SF-36 and the McCormick scale (p = 0.003), the Aminoff-Logue gait subscale (p = 0.025), the Aminoff-Logue micturition subscale (p = 0.013), and the Aminoff-Logue defecation subscale (p = 0.004). Reliability was demonstrated for all 8 SF-36 domain scales and the Physical Component Summary and the Mental Component Summary of the SF-36, where in each the Cronbach alpha satisfied the Nunnally criterion of > 0.85. CONCLUSIONS: The authors' results demonstrated that SF-36 provides valid and reliable data for patients with IPSTs and that the survey can be used appropriately to evaluate these patients.

Screening of the inhibitory effect of xenobiotics on alcohol metabolism using S9 rat liver fractions.

The purpose of this work was to develop and optimize a simple and suitable method to detect the potential inhibitory effect of drugs and medicines on alcohol dehydrogenase (ADH) activity in order to evaluate the possible interactions between medicines and alcohol metabolism. Commonly used medicines that are often involved in court litigations related with driving under the influence of alcohol were selected. Alprazolam, flunitrazepam and tramadol were tested as drugs with no known effect on ADH activity. Cimetidine, reported previously as having inhibitory effect on ADH, and 4-methylpyrazole (4-MP), a well known ADH inhibitor, were tested as positive controls. Apart from 4-MP, tramadol was identified as having the higher inhibitory effect with an IC50 of 44.7 × 10(-3)mM, followed by cimetidine (IC50 of 122.9 × 10(-3)mM). Alprazolam and flunitrazepam also reduced liver ADH activity but to a smaller extent (inhibition of 11.8 ± 5.0% for alprazolam 1.0mM and 34.5 ± 7.1% for flunitrazepam 0.04 mM). Apart from cimetidine, this is the first report describing the inhibitory effect of these drugs on ethanol metabolism. The results also show the suitability of the method to screen for inhibitory effect of drugs on ethanol metabolism helping to identify drugs for which further study is justified.

Validated method for the determination of misoprostol acid in whole blood by ultra performance liquid chromatography-tandem mass spectrometry.

Misoprostol is a pharmaceutical synthetic compound, analog of prostaglandin E1, frequently used as an abortifacient in not medically supervised or self-induced abortions, particularly in countries with restrictive abortion laws representing a serious public health problem. The aim of this study was to develop and validate a sensitive analytical method for the determination of misoprostol acid in whole blood samples. The samples were prepared by SPE and the chromatographic separation was performed by UPLC-MS/MS using ESI- and MRM mode with an Acquity UPLC(®) BEH C18 (50mm×2.1mm i.d., 1.7µm) column using a methanol-ammonium 0.1% solution gradient in a total run time of 7.0min. The method showed to be selective and linear in range 25-2000ng/L. The LOD and LOQ were 10ng/L and 25ng/L, respectively. The recovery ranged from 89 to 97%. No carryover and significant matrix effect were observed. The intra- and inter-assay precisions and the inter-assay accuracy results were 4.0% and 5.4%, 5.5% and 4.1%, and -1.4% and -2.8%, for the concentrations 50 and 500ng/L, respectively. The method developed allows the analysis of misoprostol acid in whole blood samples with adequate sensitivity to the concentration range obtained from therapeutic doses. The method was successfully used in a controlled misoprostol administration study and has been applied in our laboratory in the forensic toxicology field.