Synthesis of a PbTx-2 photoaffinity and fluorescent probe and an alternative synthetic route to photoaffinity probes

A natural phenomenon characterized by dense aggregations of unicellular photosynthetic marine organisms has been termed colloquially as red tides because of the vivid discoloration of the water. The dinoflagellate Karenia brevis is the cause of the Florida red tide bloom. K. brevis produces the brevetoxins, a potent suite of neurotoxins responsible for substantial amounts of marine mammal and fish mortalities. When consumed by humans, the toxin causes Neurotoxic Shellfish Poisoning (NSP). The native function of brevetoxin within the organism has remained mysterious since its discovery. There is a need to identify factors which contribute to and regulate toxin production within K. brevis. These toxins are produced and retained within the cell implicating a significant cellular role for their presence. Localization of brevetoxin and identification of a native receptor may provide insight into its native role as well as other polyether ladder type toxins such as the ciguatoxins, maitotoxins, and yessotoxins. In higher organisms these polyether ladder molecules bind to transmembrane proteins with high affinity. We anticipated the native brevetoxin receptor would also be a transmembrane protein. Photoaffinity labeling has become increasingly popular for identifying ligand receptors. By attaching ligands to these photophors, one is able to activate the molecule after the ligand binds to its receptor to obtain a permanent linkage between the two. Subsequent purification provides the protein with the ligand directly attached. A molecule that is capable of fluorescence is a fluorophore, which upon excitation is capable of re-emitting light. Fluorescent labeling uses fluorophores by attaching them covalently to biologically active compounds. The synthesis of a brevetoxin photoaffinity probe and its application in identifying a native brevetoxin receptor will be described. The preparation of a fluorescent derivative of brevetoxin will be described and its use in localizing the toxin to an organelle within K. brevis. In addition, the general utility of a synthesized photoaffinity label with other toxins having similar functionality will be described. An alternative synthetic approach to a general photoaffinity label will also be discussed whose goal was to accelerate the preparation and improve the overall synthetic yields of a multifunctional label.

Comprometimento Organizacional, Autocriticismo e Autocompaixão na PSP

Introdução: São poucos os estudos que em Portugal se dedicaram a explorar constructos como o autocriticismo, autocompaixão, saúde mental e o comprometimento organizacional. São nossos objetivos analisar estas associações na Polícia de Segurança Pública, considerando igualmente a influência de variáveis sociodemográficas e profissionais, bem como realizar análises preditivas que permitam verificar quais são as variáveis preditores do comprometimento organizacional. Metodologia: A amostra é constituída por 188 polícias com idades compreendidas entre os 30 e 56 anos (M = 43,75, DP = 5,66). Os participantes preencheram um protocolo integrante do projeto Organizar Positivamente do Instituto Superior Miguel Torga, composto por um questionário sociodemográfico, a SELFCS (Escala da Autocompaixão), a FSCRS (Escala das Formas do Autocriticismo e Autotranquilização), o MHI-5 (Mental Health Inventory) e o QCO (Questionário do Comprometimento Organizacional). Resultados: Os polícias parecem regular a sua relação com a organização através do comprometimento calculativo. A autocompaixão associa-se positivamente ao comprometimento afetivo, sobressaindo a capacidade de mindfulness. O autocriticismo, na forma eu inadequado - menos corrosivo que a forma eu detestado -, revela uma relação positiva com o comprometimento calculativo, pelo que a permanência na organização pode ser emocionalmente negativa, originar ataques ao eu e diminuir a capacidade afetiva. Porém, a capacidade autotranquilizadora (forma eu tranquilizador do autocriticismo) parece capacitar os polícias para o desenvolvimento de maiores níveis de comprometimento organizacional e afetivo. A idade, a antiguidade e menos habilitações literárias parecem favorecer o comprometimento calculativo. Discussão: A autocompaixão pode contribuir para uma atitude positiva e afetuosa do polícia para com a organização. Assim, através de uma intervenção baseada na autocompaixão e no mindfulness, com impacto positivo no comportamento afetivo e normativo, não só se poderá preservar e promover a saúde e o bem-estar dos polícias, mas também fomentar o empenho organizacional, atenuando os efeitos negativos da profissão policial. Esta abordagem poderá contribuir para os objetivos organizacionais, através da supressão das emoções aversivas, da diminuição da intenção de turnover e abandono da organização. / Introduction: There are few studies in Portugal that have been dedicated to explore constructs such as self-criticism, self-compassion, mental health and organizational commitment. Our goals are to analyze these associations at the Public Safety Police, considering also the influence of sociodemographic and professionals variables, as well as to conduct predictive analyses, in order to verify which variables may predict organizational commitment. Methodology: The sample consisted of 188 police with ages between 30 and 56 years (M = 43.75, SD = 5.66). The participants filled in a protocol from the project Organizar Positivamente, from Instituto Superior Miguel Torga, composed by a sociodemographic questionnaire, the SELFCS (Self-Compassion Scale), the FSCRS (Scale of Forms of Self-Criticism and Self-Reassurance, the MHI-5 (Mental Health Inventory) and the QCO (Organizational Commitment Questionnaire). Results: The police officers appear to regulate their relationship with the organization through calculative commitment. Self-compassion is associated positively to affective commitment, with mindfulness showing to be the most important competence in this professional group. Self-criticism, in its inadequate self form – which is less corrosive than the hated self form - reveals a positive relationship with calculative commitment, which might imply that staying in the organization may be emotionally negative, may provoke attacks against the self and diminish the affective capacity. However, the self reassurance capacity (reassure self form of self-criticism) may enable the police officers to develop higher levels of organizational commitment and affective commitment, as well. Age, seniority and fewer qualifications seem to support the calculative commitment. Discussion: Through self-compassion police officers may experience a positive and warmth attitude towards the organization. Thus, an intervention program based on self-compassion and mindfulness may stimulate affective and normative commitment, not only preserving and promoting health and the well-being of police officers, but also fostering organizational commitment and mitigating negative effects in the police profession. This approach may contribute to organizational goals’ through the suppression of aversive emotions, decreasing turnover and intention to abandon the organization.

Estudo dos Sintomas Psicopatológicos nas Forças Policiais Portuguesas: estudo comparativo na GNR, PSP e PJ

O presente estudo exploratório visa analisar a possibilidade de existência de Sintomas Psicopatológicos nas Forças Policiais Portuguesas GNR, PSP e PJ. Por se verificar um aumento do suicídio, nestes profissionais, com uma taxa quatro vezes superior no ano de 2008. O objectivo é essencialmente o de prevenção, conhecendo as causas, poder-se-ia intervir em tempo próprio diminuindo a taxa de suicídio. Para atingir o objectivo enunciado vamos proceder à revisão da literatura e à descrição do estudo. Foram utilizados o questionário sócio-demográfico e o BSI (Brief Symptoms Inventory) na recolha dos dados de 45 polícias (15 GNR, 15 PSP e 15 PJ) que exercem funções na Zona Centro do País e que voluntariamente colaboraram. Uma vez recolhidos os dados, estes foram introduzidos e processados no SPSS (Statistical Package for the Social Sciences) versão 13.0. Os resultados obtidos permitiram concluir que não existem diferenças estatisticamente significativas nas diferentes Forças policiais, relativamente aos sintomas Psicopatológicos. / This present exploratory study pretends to analyze the existence of psicopatologic symptoms in the Portuguese Policies Forces, Republican National Guard, Public Security Police and Judicial Police. By the constatation of the increase of the suicidal tendence in this professionals, with a four time superior tax when compared with 2008. The essential objective is the prevention of suicide, well known their causes, we will can act in the own an favorable timing diminishing the tax of suicidal tentative. To obtain the pronounced objective, we will proceed to a reading revision over the behaviour characteristics as the subjects that commit suicide and the role of the police activities that have over the happenings. After, that we will describe the used metodology that involve a social demographic enquire and the BSI that (Brief Symptoms Inventory) over a sample of 45 police man (15 GNR, 15 PSP and 15 PJ) wich activities are situated in the center of Portugal that have been voluntary to this experience. Once data is collected, it was entered into a computerized database and processed in the statistical program SPSS (Statistical Package for the Social Sciences). The obtained results will allow to conclude if exist significant statistical differences in the various Policies Forces, related with the psicopatologic symptoms.

High resolution mass spectrometry for quantitative analysis and untargeted screening of algal toxins in mussels and passive samplers

Measurement of marine algal toxins has traditionally focussed on shellfish monitoring while, over the last decade, passive sampling has been introduced as a complementary tool for exploratory studies. Since 2011, liquid chromatography-tandem mass spectrometry (LC-MS/MS) has been adopted as the EU reference method (No.15/2011) for detection and quantitation of lipophilic toxins. Traditional LC-MS approaches have been based on low-resolution mass spectrometry (LRMS), however, advances in instrument platforms have led to a heightened interest in the use of high-resolution mass spectrometry (HRMS) for toxin detection. This work describes the use of HRMS in combination with passive sampling as a progressive approach to marine algal toxin surveys. Experiments focused on comparison of LRMS and HRMS for determination of a broad range of toxins in shellfish and passive samplers. Matrix effects are an important issue to address in LC-MS; therefore, this phenomenon was evaluated for mussels (Mytilus galloprovincialis) and passive samplers using LRMS (triple quadrupole) and HRMS (quadrupole time-of-flight and Orbitrap) instruments. Matrix-matched calibration solutions containing okadaic acid and dinophysistoxins, pectenotoxin, azaspiracids, yessotoxins, domoic acid, pinnatoxins, gymnodimine A and 13-desmethyl spirolide C were prepared. Similar matrix effects were observed on all instruments types. Most notably, there was ion enhancement for pectenotoxins, okadaic acid/dinophysistoxins on one hand, and ion suppression for yessotoxins on the other. Interestingly, the ion selected for quantitation of PTX2 also influenced the magnitude of matrix effects, with the sodium adduct typically exhibiting less susceptibility to matrix effects than the ammonium adduct. As expected, mussel as a biological matrix, quantitatively produced significantly more matrix effects than passive sampler extracts, irrespective of toxin. Sample dilution was demonstrated as an effective measure to reduce matrix effects for all compounds, and was found to be particularly useful for the non-targeted approach. Limits of detection and method accuracy were comparable between the systems tested, demonstrating the applicability of HRMS as an effective tool for screening and quantitative analysis. HRMS offers the advantage of untargeted analysis, meaning that datasets can be retrospectively analysed. HRMS (full scan) chromatograms of passive samplers yielded significantly less complex data sets than mussels, and were thus more easily screened for unknowns. Consequently, we recommend the use of HRMS in combination with passive sampling for studies investigating emerging or hitherto uncharacterised toxins.

Validation d’un procédé automatisé d’extraction sur phase solide de glandes digestives de moules pour l’identification et la quantification des dinophysistoxines en LC/ESI/SM2 par piégeage d’ions quadripolaire

This report presents a new extraction method of the dinophysistoxins (DTXs), confirmed by quantification using high-performance liquid chromatography coupled to mass spectrometry with an ion trap and electro spray interface (HPLC/ESI/MS2). The method originality consists on the adaptation of DTXs basic extraction procedure (liquid/ liquid) to a solid phase extraction (SPE) via a robotic station: ASPEC XLi The parameters of the automatization procedure were optimized to obtain the best DTXs recovery rate. These improvements were loaded with digestive gland mussel homogenat realized on a silica cartridge SPE, activated in hexane/chloroform (50:50), washed with hexane/chloroform (50:50) and extracted by an elution gradient (chloroform methanol (65:35) and methanol (100%)). This method was validated according to two normative referentials (linearity, detection quantification limits and accuracy…) : - The Guide of the Pharmacy industry: Analytical Validation, report of the commission SFSTP 1992 (French Corporation of the Sciences and Technical Pharmaceutical), - - The Procedure of validation of an alternative method in compare to a reference method. (AFNOR, 1998. NF V 03-110). Comparison with the classical liquid/liquid extraction and the automated method present clear advantages. In an analytical method the extraction is generally considered to be the most labor-intensive and error-prone step. This new procedure allowed us to increase throughput, to improve the reproducibility and to reduce the error risks due to the individual manual treatments.

Screening for childhood lead poisoning in the industrial region of Fez, Morocco

International audience

Hematobiochemical changes of lead Poisoning and amelioration with Coconut ( Cocos nucifera L.) Water in wistar albino rats

To determine the ameliorative effect of coconut water on haematobiochemical changes due to lead poisoning in wistar albino rats for six weeks, sixty rats were assigned to four groups. 0.10g/l of lead and 75ml coconut ( cocus nucifera l.) water were given orally for six weeks. The mean values of red blood cells, mean corpuscular volume, mean corpuscular haemoglobin, mean corpuscular haemoglobin concentration, red blood cell distribution width and platelets (8.10±0.63(×106μl), 52.7±0.87(μm3), 17.9±0.56(pg), 34.73±0.65(g/dl), 17.90±0.67(%) and 670.00±42.22(×103μl) respectively) reduced in lead treated rats when compared with control mean values (8.41 ± 0.90(×106μl), 56.60 ± 1.55(μm3), 19.33 ± 0.82(pg), 34.93 ± 0.90(g/dl), 18.27 ± 0.73(%) and 818.33± 123.68(×103μl) respectively ) and these values increased in75ml coconut water only group and the group of 0.10g/l lead + 75ml coconut water except mean corpuscular haemoglobin, mean corpuscular haemoglobin concentration, and red blood cell distribution width of the 75ml coconut water only. The mean values of white blood cells, lymphocytes, total cholesterol, triglyceride, high density lipoprotein-cholesterol, low density lipoprotein-cholesterol, low density lipoprotein-cholesterol/high density lipoproteincholesterol and total cholesterol/high density lipoprotein-cholesterol increased (12.23±0.57(×103μl), 79.83±3.87(%), 64.66±6.01(mg/dl), 89.00±7.94(mg/dl), 22.67±6.93(mg/dl), 21.00±4.58(mg/dl), 1.29±0.62 and 3.36±0.83 respectively ) in the lead group when compared with mean values of control group (5.83±0.74(×103μl), 69.07±10.57(%), 54.00±4.04(mg/dl), 97.33±11.34(mg/dl), 20.00±3.06(mg/dl), 17.00±6.51(mg/dl), 0.97±0.41 and 2.87±0.55 respectively) but the mean values decreased when compared with the mean values of group of 75ml coconut water only and group of 0.10g/l lead + 75ml coconut water, except the mean values of high density lipoprotein-cholesterol. These results indicate that coconut water could ameliorate effects of lead toxicity

Clinical Signs and Symptoms and Laboratory Findings of Methadone Poisoning in Children

Background: Poisoning accounts for about 7% of all accidents in children under 5 years and is implicated in over 5% of all childhood deaths in developing countries. Objectives: Due to the potential risks of methadone poisoning in children and increased cases of methadone poisoning among Iranian children, this study was conducted to investigate the clinical signs and symptoms and laboratory findings of methadone toxicity in children. Patients and Methods: The present retrospective, descriptive, cross-sectional study describes the clinical symptoms and signs and laboratory findings of methadone poisoning in children under 12 years old in Shahid Beheshti Hospital, Kashan, during the years 2009 to 2013. Results: Of 58 patients, 33 (56.9%) were male and 25 (43.1%) female (P = 0.294). The mean age of patients was 5.2 ± 1.0 years. All the cases of poisoning happened with methadone syrup, due to unsafe keeping of methadone in mineral water bottles and containers of other drugs. Signs and symptoms included drowsiness (91.4 %), miosis (75.9%), vomiting (69.0%), ineffective breathing (any kind of breathing problem except apnea) (62.1%), apnea (53.4%), cyanosis (43.1%), seizure (8.6%), ataxia (6.9%) and delirium (3.4%). Conclusions: Keeping methadone in appropriate containers and warning methadone consumers about the dangerous side effects of its consumption and the symptoms of methadone poisoning in children may minimize the occurrence of this form of poisoning and its complications in children.

In vitro preliminary hemolytic studies of toxins from jellyfish Catostylus tagi

Poster presented at the 1st CiiEM Congress: "From Basic Sciences to Clinical Research". 27-28 November 2015, Egas Moniz, Caparica, Portugal

Relationship between super antigenicity, antimicrobial resistance and origin of Staphylococcus aureus isolated

Introduction: Staphylococcus aureus is a pathogen that causes food poisoning as well as hospital and community acquired infections. Objective: Establish the profile of superantigen genes among hospital isolates in relation to clinical specimen type, susceptibility to antibiotics and hospital or community acquisition. Methods: Eighty one isolates obtained from patients at Colombian hospital, were classified by antimicrobial susceptibility, specimen type and hospital or community acquired . The PCR uniplex and multiplex was used for detection of 22 superantigen genes (18 enterotoxins, tsst-1 and three exfoliative toxins). Results: Ninety five point one percent of isolates harbored one or more of the genes with an average of 5.6 genes. Prevalence of individual genes was variable and the most prevalent was seg (51.9%). Thirty nine genotypes were obtained, and the genotype gimnou (complete egc cluster) was the most prevalent alone (16.0%) and in association with other genes (13.6%). The correlation between presence of superantigens and clinical specimen or antimicrobial susceptibility showed no significant difference. But there was significant difference between presence of superantigens and the origin of the isolates, hospital or community acquired (p= 0.049). Conclusions: The results show the variability of the superantigen genes profile in hospital isolates and shows no conclusive relationship with the clinical sample type and antimicrobial susceptibility, but there was correlation with community and hospital isolates. The analysis of the interplay between virulence, epidemic and antibiotic resistance of bacterial populations is needed to predict the future of infectious diseases.

First report of Ciguatoxins in two starfish species: Ophidiaster ophidianus and Marthasterias glacialis

Ciguatera fish poisoning (CFP) is a syndrome caused by the ingestion of fish contaminated with Ciguatoxins (CTXs). These phycotoxins are produced mainly by dinoflagellates that belong to the genus Gambierdiscus that are transformed in more toxic forms in predatory fish guts, and are more present in the Indo-Pacific and Caribbean areas. It is estimated that CFP causes per year more than 10,000 intoxications worldwide. With the rise of water temperature and anthropogenic intervention, it is important to study the prevalence of CFP in more temperate waters. Through inter- and subtidal sampling, 22 species of organisms were collected, in Madeira and Azores archipelagos and in the northwestern Moroccan coast, during September of 2012 and June and July of 2013. A total of 94 samples of 22 different species of bivalves, gastropods, echinoderms and crustaceans where analyzed by Ultra Performance Liquid Chromatography-Mass Spectometry-Ion Trap-Time of Flight (UPLC-MS-IT-TOF) and Ultra Performance Chromatography- Mass Spectrometry (UPLC-MS). Our main aim was to detect new vectors and ascertain if there were some geographical differences. We detected for the first time putative CTXs in echinoderms, in two starfish species—M. glacialis and O. ophidianus. We detected differences regarding uptake values by organisms and geographical location. Toxin amounts were significant, showing the importance and the need for continuity of these studies to gain more knowledge about the prevalence of these toxins, in order to better access human health risk. In addition, we suggest monitoring of these toxins should be extended to other vectors, starfish being a good alternative for protecting and accessing human health risk.

New invertebrate vectors of Okadaic acid from the North Atlantic Waters - Portugal (Azores and Madeira) and Morocco

Okadaic acid and its analogues are potent phosphatase inhibitors that cause Diarrheic Shellfish Poisoning (DSP) through the ingestion of contaminated shellfish by humans. This group of toxins is transmitted worldwide but the number of poisoning incidents has declined over the last 20 years due to legislation and monitoring programs that were implemented for bivalves. In the summer of 2012 and 2013, we collected a total of 101 samples of 22 different species that were made up of benthic and subtidal organisms such echinoderms, crustaceans, bivalves and gastropods from Madeira, São Miguel Island (Azores archipelago) and the northwestern coast of Morocco. The samples were analyzed by UPLC-MS/MS. Our main objective was to detect new vectors for these biotoxins. We can report nine new vectors for these toxins in the North Atlantic: Astropecten aranciacus, Arbacia lixula, Echinaster sepositus, Holothuria sanctori, Ophidiaster ophidianus, Onchidella celtica, Aplysia depilans, Patella spp., and Stramonita haemostoma. Differences in toxin contents among the species were found. Even though low concentrations were detected, the levels of toxins that were present, especially in edible species, indicate the importance of these types of studies. Routine monitoring should be extended to comprise a wider number of vectors other than for bivalves of okadaic acid and its analogues.