Symbiont diversity is not involved in depth acclimation in the Mediterranean sea whip Eunicella singularis

In symbiotic cnidarians, acclimation to depth and lower irradiance can involve physiological changes in the photosynthetic dinoflagellate endosymbiont, such as increased chlorophyll content, or qualitative modifications in the symbiont population in favour of better adapted strains. It has been argued that a lack of capacity to acquire new symbionts could limit the bathymetric distribution of the host species, or compromise its long-term survival in a changing environment. But is that always true? To address this question, we investigated the symbiont genetic diversity in Eunicella singularis, a Mediterranean sea whip species with a wide bathymetric distribution (10 to 50 m depth), which has recently suffered from mass mortalities after periods of abnormally high sea temperatures. We measured symbiont population densities and chlorophyll content in natural populations, and followed the response of the holobionts after reciprocal transplantations to deep and shallow depths. A total of 161 colonies were sampled at 2 depths (10 and 30 m) at 5 sites in the northwestern Mediterranean. All colonies harboured a single ribosomal Symbiodinium clade (A'), but a relatively high within-clade genetic diversity was found among and within colonies. This diversity was not structured by depth, even though the deeper colonies contained significantly lower population densities of symbionts and less chlorophyll. We did, however, reveal host-symbiont specificity among E. singularis and other Mediterranean cnidarian species. Transplantation experiments revealed a limit of plasticity for symbiont population density and chlorophyll content, which in turn questions the importance of the trophic role of Symbiodinium in E. singularis

Simultaneous identification and quantitative determination in urine of the more significant metabolites of synthetic cannabinoids JWH-018, JWH-073, JWH-122 and JWH-250 using authentic references and deuterated isotopologues as internal standards

Introduction Synthetic cannabinoids (SC) are substances displaying a high affinity for cannabinoid receptor CB1 and represent the psychoactive agents in herbal mixtures called \u201cSpice\u201d or \u201cK2\u201d which are sold as an incense or smoking material mainly through the Internet. Because of its great abusive potential, several SC are banned in many countries, but despite this, the widespread use of herbal smoking mixtures containing SC may be partially explained by the fact that post-ingestion urines are known to produce negative results in standard toxicological screening methods for cannabis. As a consequence, an increasing number of analytical methods have been developed in forensic and doping control laboratories to enable the detection of illegal intake of these psychoactive substances in human fluids originating from psychiatric patients, emergency units or assessment of fitness to drive. Due to rapid metabolic transformation, the native SC are not usually detectable in urine samples and then the analytical methods must be based on the identification and quantization of their metabolites. Aims The aim of our study is to set-up, using synthesized reference standards, a LC-MS/MS method for routine screening procedures to assess the assumption of JWH-018, JWH-073, JWH 122 and JWH-250, the SC included in Table 1 of narcotic and psychotropic substances banned in Italy. The method gives the simultaneous identification of the three more significant metabolites of each cannabinoid and adequate sensitivity, precision and accuracy are assured by the use of deuterated internal standards. Methods For each of the four cannabinoids were synthesized the three more significant metabolites, the \u3c9- and (\u3c9-1)-hydroxyl and the \u3c9-carboxyl derivatives (\u3c9 position represent the terminal carbon of the N-alkyl side chain) while as internal standards were synthesized the (\u3c9-1)-hydroxyl metabolites trideuterated on the terminal methyl of the side chain. Urine samples were subjected to deconjugation using 30% hydrochloric acid at 90-95\ub0C for 60 min, followed by a solvent extraction procedure with n-hexane-ethyl acetate (9/1 v/v). The LC-MS/MS analysis was performed in positive mode on an API 4000 Triple Quadrupole Mass Spectrometer (AB Sciex) equipped with a 1,8\ub5m Acquity C-18 HSS T3 100 x 2 mm HPLC column (Waters) with isocratic elution (55 % of 10 mM HCOONH4 in water containing 0.1% HCOOH and 45 % of acetonitrile) at 45 \ub0C and at flow rate of 0.4 mL/min. Two transitions in \u2018multiple reaction monitoring\u2019 mode and the retention time have permitted the unambiguous identification of each metabolite which, through the presence of a suitable internal standard, was quantified. Result and discussion All the synthesized compounds were fully characterized with regard to the structure and purity, by 1H,13C NMR and GC-MS (after esterification with CH2N2 for the \u3c9-carboxylic metabolites). For sample treatment, the recovery of the metabolites was evaluated at different pH (1, 3, 5, 9 and 10). The validation of the method was performed testing linearity (0.5-100 ng/mL), reproducibility and accuracy (ranged between -15% and + 15%) at three levels. The method developed was applied to the analysis of urine samples from individuals who have taken SC. This LC-MS/MS method can be used for routine screening of urine specimens from subjects suspected of using \u201cherbal incense\u201d or \u201cSpice\u201d products spiked with the synthetic cannabinoids JWH-018, JWH-073, JWH 122 or JWH-250

GHB : problematiche in emergency toxicology

Le attuali problematiche della tossicologia dell'emergenza relativa al GHB sono connessi a due aspetti tra essi complementari. Il primo \ue8 quello relativo agli aspetti clinici e, in particolare, nell'obiettivazione di segni nel paziente che giunge al P.S. e che possono indirizzare verso una diagnosi d'intossicazione acuta da GHB. Infatti, la condivisione con altre sostanze (alcool, stupefacenti, farmaci psicoattivi) degli effetti sia sul SNC, sia a livello cardiovascolare rende pressocch\ue8 impossibile, in assenza del dato di laboratorio, una diagnosi clinica differenziale. Per contro, la scarsa conoscenza sui molteplici usi di questa sostanza( scopi voluttuari, doping e come mezzo disabilitante di terzi) e sulla sua diffusione, fa s\uec che il GHB non rientri nel pannello delle sostanze che vengono richieste al laboratorio. Il secondo aspetto \ue8 quello analitico ed \ue8 rappresentato dal fatto che attualmente gli screening che vengono applicati in emergenza nella tossicologia clinica non lo comprendono. Solamente l'applicazione di processi analitici con tecniche gascromatografiche accoppiate alla spettrometria di massa permette di determinare il GHB nei liquidi biologici, ma con tempi di produzione del risultato non rapidi

Abuso alcolico : indicatori analitici e loro diverso ruolo diagnostico - EtG : casistica ed esperienze applicative

Viene presentata la casistica della Sezione di Tossicologia Forense dell\u2019Universit\ue0 degli Studi di Milano che, nell\u2019ambito della collaborazione con le Commissioni Mediche Locali di Milano, dal 2010 ha ampliato la sua attivit\ue0 con la determinazione dell\u2019EtG in matrice pilifera. La casistica presentata riguarda due anni di attivit\ue0 e complessivamente si compone di 2200 casi relativi ad una selezione operata direttamente dalle CML che ha stabilito, quale necessit\ue0 di approfondimento diagnostico, una delle seguenti condizioni: 1) CDT% non determinabile per l\u2019esistenza di varianti genetiche; 2) recidiva; 3) titolarit\ue0 di patenti di categoria superiore; 4) violazione dell\u2019art. 186 C.d.S. per etilometria superiore a 1,5 g/L. Oltre a questi sono stati esaminati 56 soggetti inviati dai NOA di Milano e provincia. Quale matrice pilifera sono stati considerati solamente i capelli ed il prelievo \ue8 stato eseguito solamente per lunghezze non inferiori a 3 cm. Le determinazioni dell\u2019EtG sono state eseguite con tecnica LC/MS/MS sull\u2019idrolisato idro-metanolico, in presenza dello standard interno (EtG d5). Per la valutazione della positivit\ue0 si \ue8 fatto riferimento al valore di 30 pg/mg proposto dal Consensus on Alcohol Markers della SoH

L'evoluzione nel contenuto di principi attivi nella Cannabis, le attuali coltivazioni forzate o geneticamente modificate

A partire dalla met\ue0 degli anni \u201990 del secolo scorso, la diffusione del commercio on-line di semi selezionati di Cannabis ha condotto alla proliferazione di coltivazioni domestiche con produzione di piante ad elevato tenore di delta-9-THC. Verosimilmente, gli intermediari del traffico internazionale hanno, per ovvii motivi economici, incentivato l\u2019utilizzo della tecnologia OGM migliorando la resa delle coltivazioni e immettendo sul mercato dei derivati (marijuana) di qualit\ue0 superiore. Sulla base dei dati chimico-tossicologici, derivanti da osservazioni personali, gli Autori delineano il profilo delle concentrazioni dei Cannabinoli nei campioni vegetali di piccola e media entit\ue0 sequestrati in diverse realt\ue0 territoriali del Nord Italia

Fetal and perinatal desths: drugs exposure duruing pregancy

The presentation of this review will focus attention on the area of pediatric forensic medicine and the importance of through investigation of these deaths not only for the appropriate classification of the cause adn manner of deaths, but for complete documentation of toxicological data and pathological findings wich may provide essential informations about the circumstances surrounding the death and possible contributory mechanism

Contributo chimico-tossicologico alla diagnosi post-mortale di chetoacidosi metabolica

Postmortem diagnosis of diabete ketoacidosis is a complex process. As suggest by the specialist literature, circumstantial data must be supported by toxicological and chemical findings. The Authors describe two cases of death in insulin-dependent men where the autopsy revealed no significant findings in determining the causes of death. Blood and urine werw submitted for glucose analysis and showed for both cases level exceeding 500 mg/dL. No drugs were detected in toxicological screening. Glucose was also confirmed on pretreated samples of blood by fully-scan gaschromatography -mass spectrometry (GC/MS) with a previous derivation with BSTFA. The femoral blood samples of each case were divided in two portions. One of these was preserved in a glass tube with sodium fluoride and investigated for volatile compounds by headspace gaschromatographic technique. Acetone and isopropanol have been detected in each case. No ethanol was detected. The second portion of femoral blood was placed in a glass tube without preservative, stored at 4\ub0C and analysed two months later for volatile compounds and glucose. The analysis showed for both cases the absence of glucose and the presence of ethanol. That can attributed to postmortem phenomena. This cases report point out as the postmortem diagnosis of diabetic ketoacidosis can be supported by the chemical and toxicological investigations, above all when the antemortem clinical data and suitable biological samples are not available