(Table 1) Boron concentration and d11B values of interstitial water from IODP Holes 308-U1319A, U1320A, U1322B and U1324B

The Integrated Ocean Drilling Program Expedition 308 (IODP308) drilled normal-pressured sediments from the Brazos-Trinity Basin IV and over-pressured sediments from the Ursa Basin on the northern slope of the Gulf of Mexico. The interstitial water samples from the normal-pressured basin show B concentrations and B isotopic compositions ranging from 255 to 631 µM (0.6 to 1.5 times of seawater value) and from +29.1 to +42.7 per mil (relative to NIST SRM 951), respectively. A wider range is observed both for B concentrations (292 to 865 µM, 0.7 to 2.1 times of seawater value) and d11B values (+25.5 to +43.2 per mil) of the interstitial water in the over-pressured basin. The down-core distribution of B concentrations and d11B values in the interstitial waters are sensitive tracers for assessing various processes occurring in the sediment column, including boron adsorption/desorption reactions involving clay minerals and organic matter in sediments as well as fluid migration and mixing in certain horizons and in the sediment column. In the normal-pressured basin adsorption/desorption reactions in shallow sediments play the major role in controlling the B content and B isotopic composition of the interstitial water. In contrast, multiple processes affect the B content and d11B of the interstitial water in the over-pressured Ursa Basin. There, the stratigraphic level of the maxima of B and d11B correspond to seismic reflectors. The intruded fluids along the seismic reflector boundary from high to low-topography mix with local interstitial water. Fluid flow is inferred in the Blue Unit (a coarse sandstone layer, connecting the high- to low-pressured region) from the freshening of interstitial water in Ursa Basin Site U1322, and upward flow by the overpressure expels fluid from the overburden above the Blue Unit.

Fossilization and degradation of archaeal intact polar tetraether lipids in ODP Site 201-1229 sediments

Glycerol dibiphytanyl glycerol tetraether (GDGT) lipids are part of the cellular membranes of Thaumarchaeota, an archaeal phylum composed of aerobic ammonia oxidizers, and are used in the paleotemperature proxy TEX86. GDGTs in live cells possess polar head groups and are called intact polar lipids (IPL-GDGTs). Their transformation to core lipids (CL) by cleavage of the head group was assumed to proceed rapidly after cell death but it has been suggested that some of these IPL-GDGTs can, just like the CL-GDGTs, be preserved over geological timescales. Here, we examined IPL-GDGTs in deeply buried (0.2-186 mbsf, ~2.5 Myr) sediments from the Peru Margin. Direct measurements of the most abundant IPL-GDGT, IPL-crenarchaeol, specific for Thaumarchaeota, revealed depth profiles which differed per head group. Shallow sediments (<1 mbsf) contained IPL-crenarchaeol with both glycosidic- and phosphate headgroups, as also observed in thaumarchaeal enrichment cultures, marine suspended particulate matter and marine surface sediments. However, hexose, phosphohexose-crenarchaeol is not detected anymore below 6 mbsf (~7 kyr), suggesting a high lability. In contrast, IPL-crenarchaeol with glycosidic head groups is preserved over time scales of Myr. This agrees with previous analyses of deeply buried (>1 m) marine sediments, which only reported glycosidic and no phosphate-containing IPL-GDGTs. TEX86 values of CL-GDGTs did not markedly change with depth, and the TEX86 of IPL-derived GDGTs decreased only when the proportions of monohexose- to dihexose-GDGTs changed, likely due to the enhanced preservation of the monohexose GDGTs. Our results support the hypothesis that in situ GDGT production and differential IPL degradation in sediments is not substantially affecting TEX86 paleotemperature estimations based on CL GDGTs and indicate that likely only a small amount of IPL-GDGTs present in deeply buried sediments is part of cell membranes of active Archaea. The amount of archaeal biomass in the deep biosphere based on these IPLs may have been substantially overestimated.

The Responsiveness, Content Validity, and Convergent Validity of the Measure Yourself Concerns and Wellbeing (MYCaW) Patient-Reported Outcome Measure

Objective Measure Yourself Concerns and Wellbeing (MYCaW) is a patient-centred questionnaire that allows cancer patients to identify and quantify the severity of their ‘Concerns’ and Wellbeing, as opposed to using a pre-determined list. MYCaW administration is brief and aids in prioritising treatment approaches. Our goal was to assess the convergent validity and responsiveness of MYCaW scores over time, the generalisability of the existing qualitative coding framework in different complementary and integrative healthcare settings and content validity. Methods Baseline and 6-week follow-up data (n=82) from MYCaW and FACIT-SpEx questionnaires were collected for a service evaluation of the ‘Living Well With The Impact of Cancer’ course at Penny Brohn Cancer Care. MYCaW construct validity was determined using Spearman's Rank Correlation test, and responsiveness indices assessed score changes over time. The existing qualitative coding framework was reviewed using a new dataset (n=158) and coverage of concern categories compared to items of existing outcome measures. Results Good correlation between MYCaW and FACIT-SpEx score changes were achieved (r= -0.57, p≥0.01). MYCaW Profile and Concern scores were highly responsive to change: SRM=1.02 and 1.08; effect size=1.26 and 1.22. MYCaW change scores showed the anticipated gradient of change according to clinically relevant degrees of change. Categories including ‘Spirituality’, ‘weight change’ and ‘practical concerns’ were added to the coding framework to improve generalisability. Conclusions MYCaW scores were highly responsive to change, allowing personalized patient outcomes to be quantified; the qualitative coding framework is generalisable across different oncology settings and has broader coverage of patient-identified concerns compared with existing cancer-related patient-reported outcome measures.

Measuring the Diversity and the Integration of Film Production in Mercosur: A Social Network Analysis

Based on an original and comprehensive database of all feature fiction films produced in Mercosur between 2004 and 2012, the paper analyses whether the Mercosur film industry has evolved towards an integrated and culturally more diverse market. It provides a summary of policy opportunities in terms of integration and diversity, emphasizing the limiter role played by regional policies. It then shows that although the Mercosur film industry remains rather disintegrated, it tends to become more integrated and culturally more diverse. From a methodological point of view, the combination of Social Network Analysis and the Stirling Model opens up interesting research tracks to analyse creative industries in terms of their market integration and their cultural diversity.

Stories from the Inside:The Prisons Memory Archive

A close textual reading of the online-available recording of ex-prison officer, John Heatherington, who is filmed as he returns to the site of the Maze and Long Kesh Prison, which was the largest prison oeperating during the Troubles in Northern Ireland. Using the empty site as stimulant for his memory, John recalls the tension, violence and dark humour of the period, while also showing exceptional empathy for all those, including prisoners, who passed through the prison system.The interview is taken from the Prisons Memory Archive (www.prisonsmemoryarchive.com) and, as director of the PMA, I have a particular insight to the recording process and John's special contribution to it.

Modifications neurométaboliques et microstructurales à la suite d'une commotion cérébrale chez les athlètes féminines.

L’utilisation de méthodes d’investigation cérébrale avancées a permis de mettre en évidence la présence d’altérations à court et à long terme à la suite d’une commotion cérébrale. Plus spécifiquement, des altérations affectant l’intégrité de la matière blanche et le métabolisme cellulaire ont récemment été révélées par l’utilisation de l’imagerie du tenseur de diffusion (DTI) et la spectroscopie par résonance magnétique (SRM), respectivement. Ces atteintes cérébrales ont été observées chez des athlètes masculins quelques jours après la blessure à la tête et demeuraient détectables lorsque les athlètes étaient à nouveau évalués six mois post-commotion. En revanche, aucune étude n’a évalué les effets neurométaboliques et microstructuraux dans la phase aigüe et chronique d’une commotion cérébrale chez les athlètes féminines, malgré le fait qu’elles présentent une susceptibilité accrue de subir ce type de blessure, ainsi qu’un nombre plus élevé de symptômes post-commotionnels et un temps de réhabilitation plus long. Ainsi, les études composant le présent ouvrage visent globalement à établir le profil d’atteintes microstructurales et neurométaboliques chez des athlètes féminines par l’utilisation du DTI et de la SRM. La première étude visait à évaluer les changements neurométaboliques au sein du corps calleux chez des joueurs et joueuses de hockey au cours d’une saison universitaire. Les athlètes ayant subi une commotion cérébrale pendant la saison ont été évalués 72 heures, 2 semaines et 2 mois après la blessure à la tête en plus des évaluations pré et post-saison. Les résultats démontrent une absence de différences entre les athlètes ayant subi une commotion cérébrale et les athlètes qui n’en ont pas subie. De plus, aucune différence entre les données pré et post-saison a été observée chez les athlètes masculins alors qu’une diminution du taux de N-acetyl aspartate (NAA) n’a été mise en évidence chez les athlètes féminines, suggérant ainsi un impact des coups d’intensité sous-clinique à la tête. La deuxième étude, qui utilisait le DTI et la SRM, a révélé des atteintes chez des athlètes féminines commotionnées asymptomatiques en moyenne 18 mois post-commotion. Plus spécifiquement, la SRM a révélé une diminution du taux de myo-inositol (mI) au sein de l’hippocampe et du cortex moteur primaire (M1) alors que le DTI a mis en évidence une augmentation de la diffusivité moyenne (DM) dans plusieurs faisceaux de matière blanche. De iii plus, une approche par région d’intérêt a mis en évidence une diminution de la fraction d’anisotropie (FA) dans la partie du corps calleux projetant vers l’aire motrice primaire. Le troisième article évaluait des athlètes ayant subi une commotion cérébrale dans les jours suivant la blessure à la tête (7-10 jours) ainsi que six mois post-commotion avec la SRM. Dans la phase aigüe, des altérations neuropsychologiques combinées à un nombre significativement plus élevé de symptômes post-commotionnels et dépressifs ont été trouvés chez les athlètes féminines commotionnées, qui se résorbaient en phase chronique. En revanche, aucune différence sur le plan neurométabolique n’a été mise en évidence entre les deux groupes dans la phase aigüe. Dans la phase chronique, les athlètes commotionnées démontraient des altérations neurométaboliques au sein du cortex préfrontal dorsolatéral (CPDL) et M1, marquées par une augmentation du taux de glutamate/glutamine (Glx). De plus, une diminution du taux de NAA entre les deux temps de mesure était présente chez les athlètes contrôles. Finalement, le quatrième article documentait les atteintes microstructurales au sein de la voie corticospinale et du corps calleux six mois suivant une commotion cérébrale. Les analyses n’ont démontré aucune différence au sein de la voie corticospinale alors que des différences ont été relevées par segmentation du corps calleux selon les projections des fibres calleuses. En effet, les athlètes commotionnées présentaient une diminution de la DM et de la diffusivité radiale (DR) au sein de la région projetant vers le cortex préfrontal, un volume moindre des fibres de matière blanche dans la région projetant vers l’aire prémotrice et l’aire motrice supplémentaire, ainsi qu’une diminution de la diffusivité axiale (DA) dans la région projetant vers l’aire pariétale et temporale. En somme, les études incluses dans le présent ouvrage ont permis d’approfondir les connaissances sur les effets métaboliques et microstructuraux des commotions cérébrales et démontrent des effets délétères persistants chez des athlètes féminines. Ces données vont de pair avec la littérature scientifique qui suggère que les commotions cérébrales n’entraînent pas seulement des symptômes temporaires.

Interview with Charles W. Peck

Interview in five sessions, October-November 2003, with Charles W. Peck, professor of physics (now emeritus) in the Division of Physics, Mathematics, and Astronomy. He recalls his early life in South Texas and his interest in radio; first year of college at Texas Arts & Industries; three more years at New Mexico College of Agriculture & Mechanical Arts. Recalls graduate studies at Caltech with Murray Gell-Mann, H. P. Robertson, Robert Walker, Richard A. Dean, W. R. Smythe. Works on increasing intensity and stability of the Caltech synchrotron, with Walker, Matt Sands, and Alvin Tollestrup; 1964 thesis on K-lambda photoproduction. Joins the faculty as an assistant professor in 1965. Discusses his various teaching assignments, including an embarrassing moment when Richard Feynman attended one of his freshman physics lectures. Discusses his research at the Stanford Linear Accelerator Center and Lawrence Radiation Laboratory’s Bevatron. Collaboration with UC Berkeley and SLAC on “crystal ball” detector for SLAC’s SPEAR storage ring. Taking the crystal ball to DESY, in Hamburg. Works with Barry Barish at Gran Sasso laboratory in Italy, on MACRO; search for magnetic monopoles. He also discusses his administration work at Caltech, as executive officer for physics (1983-1986) and as PMA division chair from 1993 to 1998, when he immediately had to deal with the troubles plaguing LIGO [Laser Interferometer Gravitational-wave Observatory]. Detailed discussion of the LIGO contretemps and how it was settled, and of turning Big Bear Solar Observatory over to the New Jersey Institute of Technology. Advent of David Baltimore as Caltech president; attempt to recruit Ed Witten.

Kyrönjoen tekoaltaiden tila syvänteiden pohjaeläimistön perusteella : Vuoden 2015 tulokset ja tilan kehitys

Kyrönjoella on vuosina 1968–2004 toteutettu laaja tulvasuojelutyö, joka perustuu vuonna 1965 valmistuneeseen vesistötaloussuunnitelmaan. Tulvasuojelutyöhön ovat kuuluneet muun muassa Liikapuron (1966–68), Pitkämön (1968–71), Kalajärven (1971–76) ja Kyrkösjärven (1977–83) tekoaltaat, sekä näihin liittyvät täyttö- ja tyhjennysuomat, säännöstelypadot ja voimayhtiöiden rakentamat voimalaitokset. Tässä raportissa arvioidaan tekoaltaiden tilaa ja kehitystä syvänteistä otettujen pohjaeläinnäytteiden perusteella. Harvasukasmadot (Oligochaeta) ja surviaissääsket (Chironomidae) olivat tekoaltaiden syvänteiden vallitsevia pohjaeläinryhmiä. Pitkämön tekoaltaan pohjaeläinyhteisö erosi selkeästi muista järvistä vähälajisuudellaan, ja altaan pohjaeläinyhteisö koostui pääosin harvasukasmadoista. Myös Kyrkösjärvessä ja Kalajärvessä harvasukasmatojen osuus oli vuosina 2009 ja 2015 suurempi kuin surviaissääskien. Liikapuron altaassa puolestaan valtaosa pohjaeläinyhteisöstä oli surviaissääskiä. Kaikki tutkitut tekoaltaat olivat vuonna 2015 reheviä ja PMA-luokan perusteella tilaltaan välttäviä tai huonoja. PICM-indeksiä voidaan käyttää luotettavasti ainoastaan tekoaltaista syvimpään eli Pitkämöön, joka luokittui indeksin perusteella tyydyttäväksi vuonna 2015. Pohjaeläinten taksonimäärässä ei ole tapahtunut Kalajärvellä selväsuuntaista muutosta 1980-luvulta lähtien, mutta Liikapuron, Kyrkösjärven ja Pitkämön tekoaltaissa taksonimäärä oli vuosina 2009 ja 2015 selvästi suurempi kuin aiemmin. Pohjaeläinten yksilötiheydet ovat olleet Pitkämöllä 2000-luvulla ja Liikapurolla vuosina 2009 ja 2015 suuremmat kuin 1980-luvulla. Kalajärvellä ja Kyrkösjärvellä yksilötiheys oli vuonna 1986 selvästi suurempi kuin 2000-luvulla. Kyrönjoen valuma-alueen tekoaltaat muistuttavat syvännepohjaeläimistöltään reheviä luonnonjärviä. Tämä ei johdu niinkään järvien vedenlaadusta, vaan pohjan ominaisuuksista, säännöstelystä ja järvien mataluudesta. On arvioitu, että pohja-aineksen hajotus ja uusien hienojakoisten pohjasedimenttien muodostuminen kestää suomalaisissa keskihumuksisissa tekoaltaissa kymmeniä, ellei satoja vuosia. Tämän tutkimuksen perusteella pohjaeläinyhteisöjen sukkessio ja kolonisaatio etenee tekoaltaissa edelleen.

Secretaria on-line from ISCAP

Like many other higher educat ion schools, ISCAP`s population has grown at a rate of almost 100% in the end of the twentieth century. Its administrative structures were reinforced, but it was not in the same proportion. Face to face with the inability to resolve the problem, the administration decided to implement a computer based system, available in the Internet. In a first stage, in 1997, the system was implemented as a services support. The next stage, in 1999, proposes to increase student services quality. A project that aims to bring student services available on the Internet begins to be developed.

Aplicação das tecnologias de informação e comunicação como estratégia para a retenção de aunos

Este artigo aborda a aplicação das Tecnologias de Informação e Comunicação como estratégias de boas práticas que conduzem à retenção e captura de alunos do ensino superior. É baseado num estudo de caso de oito anos, de utilização de um completo sistema de informação. Este sistema, além de constituir um ERP, de suporte às actividades de gestão académica, dispõe ainda de uma forte componente de SRM que confere suporte às actividades administrativas e lectivas. É descrito em que medida o sistema apresentado facilita a interacção e comunicação entre os membros da comunidade académica, recorrendo à internet, com serviços disponíveis na Web complementando-os com correio electrónico, SMS e CTI. Através de uma percepção, sustentada por análise empírica e por resultados de inquéritos, demonstra-se como este tipo de boas práticas pode elevar o nível de satisfação da comunidade. Muito em particular, é possível combater o insucesso escolar, evitar que alunos abandonem os seus cursos antes do seu término e que os recomendem a potenciais alunos. Em complemento, este tipo de estratégia permite ainda fortes economias na gestão da instituição, elevando o seu valor. Como trabalho futuro, é apresentada a nova fase do projecto que envereda pela aplicação de Business Intelligence para optimização do processo de gestão, tornando-o pró-activo. Também é apresentada a visão tecnológica que orienta os novos desenvolvimentos, para uma arquitectura baseada em serviços Web e linguagens de definição processual.

Technology strategies in the process of student retention and efficiency of academic management

This article discusses the application of Information and Communication Technologies and strategies for best practices in order to capture and maintain faculty students' attention. It is based on a case study of ten years, using a complete information system. This system, in addition to be considered an ERP, to support the activities of academic management, also has a strong component of SRM that provides support to academic and administrative activities. It describes the extent to which the presented system facilitates the interaction and communication between members of the academic community, using the Internet, with services available on the Web complementing them with email, SMS and CTI. Through a perception, backed by empirical analysis and results of investigations, it demonstrates how this type of practice may raise the level of satisfaction of the community. In particular, it is possible to combat failure at school, avoid that students leave their course before its completion and also that they recommend them to potential students. In addition, such a strategy also allows strong economies in the management of the institution, increasing its value. As future work, we present the new phase of the project towards implementation of Business Intelligence to optimize the management process, making it proactive. The technological vision that guides new developments to a construction based on Web services and procedural languages is also presented.

The regulation of AMP-activated protein kinase by vascular endothelial growth factor

The function of the vascular endothelium is to maintain vascular homeostasis, by providing an anti-thrombotic, anti-inflammatory and vasodilatory interface between circulating blood and the vessel wall, meanwhile facilitating the selective passage of blood components such as signaling molecules and immune cells. Dysfunction of the vascular endothelium is implicated in a number of pathological states including atherosclerosis and hypertension, and is thought to precede atherogenesis by a number of years. Vascular endothelial growth factor A (VEGF) is a crucial mitogenic signaling molecule, not only essential for embryonic development, but also in the adult for regulating both physiological and pathological angiogenesis. Previous studies by our laboratory have demonstrated that VEGF-A activates AMP-activated protein kinase (AMPK), the downstream component of a signaling cascade important in the regulation of whole body and cellular energy status. Furthermore, studies in our laboratory have indicated that AMPK is essential for VEGF-A-stimulated vascular endothelial cell proliferation. AMPK activation typically stimulates anabolic processes and inhibits catabolic processes including cell proliferation, with the ultimate aim of redressing energy imbalance, and as such is an attractive therapeutic target for the treatment of obesity, metabolic syndromes, and type 2 diabetes. Metabolic diseases are associated with adverse cardiovascular outcomes and AMPK activation is reported to have beneficial effects on the vascular endothelium. The mechanism by which VEGF-A stimulates AMPK, and the functional consequences of VEGF-A-stimulated AMPK activation remain uncertain. The present study therefore aimed to identify the specific mechanism(s) by which VEGF-A regulates the activity of AMPK in endothelial cells, and how this might differ from the activation of AMPK by other agents. Furthermore, the role of AMPK in the pro-proliferative actions of VEGF-A was further examined. Human aortic and umbilical vein endothelial cells were therefore used as a model system to characterise the specific effect(s) of VEGF-A stimulation on AMPK activation. The present study reports that AMPK α1 containing AMPK complexes account for the vast majority of both basal and VEGF-A-stimulated AMPK activity. Furthermore, AMPK α1 is localized to the endoplasmic reticulum when sub-confluent, but translocated to the Golgi apparatus when cells are cultured to confluence. AMPK α2 appears to be associated with a structural cellular component, but neither α1 nor α2 complexes appear to translocate in response to VEGF-A stimulation. The present study confirms previous reports that when measured using the MTS cell proliferation assay, AMPK is required for VEGF-A-stimulated endothelial cell proliferation. However, parallel experiments measuring cell proliferation using the Real-Time Cell Analyzer xCELLigence system, do not agree with these previous reports, suggesting that AMPK may in fact be required for an aspect of mitochondrial metabolism which is enhanced by VEGF-A. Studies into the mitochondrial activity of endothelial cells have proved inconclusive at this time, but further studies into this are warranted. During previous studies in our laboratory, it was suggested that VEGF-A-stimulated AMPK activation may be mediated via the diacylglycerol (DAG)-sensitive transient receptor potential cation channel (TRPCs -3, -6 or -7) family of ion channels. The present study can neither confirm, nor exclude the expression of TRPCs in vascular endothelial cells, nor rule out their involvement in VEGF-A-stimulated AMPK activation; more specific investigative tools are required in order to characterise their involvement. Furthermore, nicotinic acid adenine dinucleotide phosphate (NAADP)-stimulated Ca2+ release from acidic intracellular organelles is not required for AMPK activation by VEGF-A. Despite what is known about the mechanisms by which AMPK is activated, far less is known concerning the downregulation of AMPK activity, as observed in human and animal models of metabolic disease. Phosphorylation of AMPK α1 Ser485 (α2 Ser491) has recently been characterised as a mechanism by which the activity of AMPK is negatively regulated. We report here for the first time that VEGF-A stimulates AMPK α1 Ser485 phosphorylation independently of the previously reported AMPK α1 Ser485 kinases Akt (protein kinase B) and ERK1/2 (extracellular signal-regulated kinase 1/2). Furthermore, inhibition of protein kinase C (PKC), the activity of which is reported to be elevated in metabolic disease, attenuates VEGF-A- and phorbol 12-myristate 13-acetate (PMA)-stimulated AMPK α1 Ser485 phosphorylation, and increases basal AMPK activity. In contrast to this, PKC activation reduces AMPK activity in human vascular endothelial cells. Attempts to identify the PKC isoform responsible for inhibiting AMPK activity suggest that it is one (or more) of the Ca2+-regulated DAG-sensitive isoforms of PKC, however cross regulation of PKC isoform expression has limited the present study. Furthermore, AMPK α1 Ser485 phosphorylation was inversely correlated with human muscle insulin sensitivity. As such, enhanced AMPK α1 Ser485 phosphorylation, potentially mediated by increased PKC activation may help explain some of the reduced AMPK activity observed in metabolic disease.

In vitro studies of drug transformations: application to forensic toxicology

The forensic toxicologist faces challenges in the detection of drugs and poisons in biological samples due to transformations which occur both during life and after death. For example, changes can result from drug metabolism during life or from the use of formalin solution for post mortem embalming purposes. The former requires the identification of drug metabolites and the latter the identification of chemical reaction products in order to know which substances had been administered. The work described in this thesis was aimed at providing ways of tackling these challenges and was divided into two parts. Part 1 investigated the use of in vitro drug metabolism by human liver microsomes (HLM) to obtain information on drug metabolites and Part 2 investigated the chemical reactions of drugs and a carbamate pesticide with formalin solution and formalin-blood. The initial aim of part I was to develop an in vitro metabolism method using HLM, based on a literature review of previous studies of this type. MDMA was chosen as a model compound to develop the HLM method because its metabolism was known and standards of its metabolites were commercially available. In addition, a sensitive and selective method was developed for the identification and quantitation of hydrophilic phase I drug metabolites using LC/MS/MS with a conventional reverse-phase (C18) column. In order to obtain suitable retention factors for polar drug metabolites on this column, acetyl derivatives were evaluated for converting the metabolites to more lipophilic compounds and an optimal separation system was developed. Acetate derivatives were found to be stable in the HPLC mobile phase and to provide good chromatographic separation of the target analytes. In vitro metabolism of MDMA and, subsequently, of other drugs involved incubation of 4 µg drug substance in pH 7.4 buffer with an NADPH generating system (NGS) at 37oC for 90 min with addition of more NGS after 30 min. The reaction was stopped at 90 min by the addition of acetonitrile before extraction of the metabolites. Acetate derivatives of MDMA metabolites were identified by LC/MS/MS using multiple reaction monitoring (MRM). Three phase I metabolites (both major and minor metabolites) of MDMA were detected in HLM samples. 3,4-dihydroxy-methamphetamine and 4-hydroxy-3-methoxymethamphetamine were found to be major metabolites of MDMA whereas 3,4-methylenedioxyamphetamine was found to be a minor metabolite. Subsequently, ten MDMA positive urines were analysed to compare the metabolite patterns with those produced by HLM. An LC/MS method for MDMA and its metabolites in urine samples was developed and validated. The method demonstrated good linearity, accuracy and precision and insignificant matrix effects, with limits of quantitation of 0.025 µg/ml. Moreover, derivatives of MDMA and its metabolites were quantified in all 10 positive human urine samples. The urine metabolite pattern was found to be similar to that from HLM. The second aim of Part 1 was to use the HLM system to study the metabolism of some new psychoactive substances, whose misuse worldwide has necessitated the development of analytical methods for these drugs in biological specimens. Methylone and butylone were selected as representative cathinones and para-methoxyamphetamine (PMA) was chosen as a representative ring-substituted amphetamine, because of the involvement of these drugs in recent drug-related deaths, because of a relative lack of information on their metabolism, and because reference standards of their metabolites were not commercially available. An LC/MS/MS method for the analysis of methylone, butylone, PMA and their metabolites was developed. Three phase I metabolites of methylone and butylone were detected in HLM samples. Ketone reduction to β-OH metabolites and demethylenation to dihydroxy-metabolites were found to be major phase I metabolic pathways of butylone and methylone whereas N-demethylation to nor-methylone and nor-butylone were found to be minor pathways. Also, demethylation to para-hydroxyamphetamine was found to be a major phase I metabolic pathway of PMA whereas β-hydroxylation to β-OH-PMA was found to be a minor pathway. Formaldehyde is used for embalming, to reduce decomposition and preserve cadavers, especially in tropical countries such as Thailand. Drugs present in the body can be exposed to formaldehyde resulting in decreasing concentrations of the original compounds and production of new substances. The aim of part II of the study was to evaluate the in vitro reactions of formaldehyde with selected drug groups including amphetamines (amphetamine, methamphetamine and MDMA), benzodiazepines (alprazolam and diazepam), opiates (morphine, hydromorphone, codeine and hydrocodone) and with a carbamate insecticide (carbosulfan). The study would identify degradation products to serve as markers for the parent compounds when these were no longer detectable. Drugs standards were spiked in 10% formalin solution and 10% formalin blood. Water and whole blood without formalin were used for controls. Samples were analysed by LC/MS/MS at different times from the start, over periods of up to 30 days. Amphetamine, methamphetamine and MDMA were found to rapidly convert to methamphetamine, DMA and MDDMA respectively, in both formalin solution and formalin blood, confirming the Eschweiler-Clarke reaction between amine-containing compounds and formaldehyde. Alprazolam was found to be unstable whereas diazepam was found to be stable in both formalin solution and water. Both were found to hydrolyse in formalin solution and to give open-ring alprazolam and open-ring diazepam. Other alprazolam conversion products attached to paraformaldehyde were detected in both formalin solution and formalin blood. Morphine and codeine were found to be more stable than hydromorphone and hydrocodone in formalin solution. Conversion products of hydromorphone and hydrocodone attached to paraformaldehyde were tentatively identified in formalin solution. Moreover, hydrocodone and hydromorphone rapidly decreased within 24 h in formalin blood and could not be detected after 7 days. Carbosulfan was found to be unstable in formalin solution and was rapidly hydrolysed within 24 h, whereas in water it was stable up to 48 h. Carbofuran was the major degradation product, plus smaller amounts of other products, 3-ketocarbofuran and 3-hydrocarbofuran. By contrast, carbosulfan slowly hydrolysed in formalin-blood and was still detected after 15 days. It was concluded that HLM provide a useful tool for human drug metabolism studies when ethical considerations preclude their controlled administration to humans. The use of chemical derivatisation for hydrophilic compounds such as polar drug metabolites for analysis by LC/MS/MS with a conventional C18 column is effective and inexpensive, and suitable for routine use in the identification and quantitation of drugs and their metabolites. The detection of parent drugs and their metabolites or conversion and decomposition products is potentially very useful for the interpretation of cases in forensic toxicology, especially when the original compounds cannot be observed.

Desenvolvimento de método empregando DLLME para extração e pré-concentração de ferro em amostras de água com diferentes salinidades

Neste trabalho foi desenvolvido e validado um método analítico para a determinação de Fe total em amostras de água com diferentes salinidades empregando a microextração líquido-líquido dispersiva (DLLME), seguido de determinação espectrofotométrica na região do ultravioleta-visível. Nas extrações por DLLME, o Fe foi complexado com pirrolidina ditiocarbamato de amônio (APDC) e extraído após a injeção de uma mistura composta de 1,2-diclorobenzeno (solvente extrator) e etanol (solvente dispersor). Os fatores que influenciam a eficiência de extração (tipo e volume de solvente extrator e dispersor, concentração de APDC e tempo de extração) e subsequente determinação espectrofotométrica foram otimizados. Após otimização das condições, a curva de calibração foi linear entre 0,02 e 2,0 mg L-1 (r 2 = 0,9998) e os limites de detecção e quantificação do método foram de 6,1 e 18,3 µg L-1 , respectivamente. O fator de pré-concentração foi de 3,3. A exatidão foi avaliada em termos de recuperação, com variação entre 90 e 102% e desvio padrão relativo (RSD) inferiores a 12%. A exatidão do método também foi avaliada utilizando um material de referência certificado (SRM 1643e, Elementos Traço em Água), o qual apresentou concordância de 104% e RSD de 5%. O método foi aplicado para amostras de água com diferentes salinidades, como água mineral, estuarina e de alto mar. Além disso, o método proposto apresentou concordância da ordem de 95%, quando comparado com o método 1,10-fenantrolina para a determinação de Fe total.

EFFECT OF SORGHUM SEED TREATMENT IN BURKINA FASO VARIES WITH BASELINE CROP PERFORMANCE AND GEOGRAPHICAL LOCATION

Sorghum [ Sorghum bicolor (L.) Moench] is a major subsistence crop throughout the region of Sahel. With the exception of seeds and labour, no agricultural inputs are in general used in sorghum production since the grain is of a relatively low commercial value and the risk of losing the crop to drought, flooding, etc. is substantial. A meta-analysis of 118 field experiments was carried out to identify conditions in which two protective seed treatments could support a yield increase of sorghum in Burkina Faso. The two treatments were: i) treatment with the pesticide Calthio C (thiram and chlorpyrifos) and ii) treatment with an aqueous extract from the plant Eclipta alba . Both treatments were found to produce a yield increase (Medians: Calthio C +199 kg ha-1, P<2x10-9; E. alba +90.5 kg ha-1 P<4x10-4). A strong relative effect of Calthio C on yield (+36%) was found for field experiments with a low baseline yield. A strong relative effect of E. alba extract on yield (+22%) was found for experiments with a low baseline of emergence. ANOVA of the 118 field tests showed that baseline crop performance (yield and emergence) and the effect of seed treatments were strongly linked to geographical location (twelve different villages included). Roots from sorghum in the village showing the strongest effect of both seed treatments (>40% yield increase) were found to carry a comparatively high load of the infectious ascomycetes: Fusarium equiseti , Macrophomina phaseolina and Curvularia lunata .