Application of Biofunctionalized Magnetic Nanoparticles Based-Sensing in Abused Drugs Diagnostics.

Real-time detection of substance use is an approach of high interest leading to the optimization of behavioral interventions and drug abuse intervention. The current methods in use suffer many limitations and need high logistical and laboratory requirements. Biosensors have shown a great potential in overcoming these limitations. In the present study, the electrochemical biosensor composed of a screen-printed electrode (SPE) was designed for the detection of synthetic cannabinoid (SC). Antibody-immobilized magnetic nanoparticles were also used to create a surface on the transducer with magnetic interactions in order to detect JWH-073 as a SC model. The use of immobilized magnetic nanoparticles to create working surfaces makes the electrode a reusable SPE which can be reutilized after the cleansing. To examine and observe any possible changes on the surface due to its interaction with the analyte, different electrochemical techniques such as differential pulse voltammetry, cyclic voltammetry, and electrochemical impedance spectrometry were applied. Based on the obtained results, the linearity of the biosensor was found between 5 and 400 ng/mL, and the detection limit was calculated as 22 ng/mL (n = 6) using the 3 Sb/m formula. The biosensor functionality was studied in the presence of some related interferents that showed lower responses than JWH-073, thus demonstrating the good selectivity of the prepared biosensor. Finally, the sensory platform was used to test synthetic urine sample, and the results were compared with obtained results from liquid chromatography quadrupole time-of-flight mass spectrometry (LC-QTOF/MS), which showed that the proposed method could be utilized to identify abuse drugs.

Active vaccination attenuates the psychostimulant effects of α-PVP and MDPV in rats.

Recreational use of substituted cathinones continues to be an emerging public health problem in the United States; cathinone derivatives α-pyrrolidinopentiophenone (α-PVP) and 3,4-methylenedioxypyrovalerone (MDPV), which have been linked to human fatalities and show high potential for abuse liability in animal models, are of particular concern. The objective of this study was to develop an immunotherapeutic strategy for attenuating the effects of α-PVP and MDPV in rats, using drug-conjugate vaccines created to generate antibodies with neutralizing capacity. Immunoconjugates (α-PVP-KLH and MDPV-KLH) or the control carrier protein, keyhole limpet hemocyanin (KLH), were administered to groups (N = 12) of male Sprague-Dawley rats on Weeks 0, 2 and 4. Groups were administered α-PVP or MDPV (0.0, 0.25, 0.5, 1.0, 5.0 mg/kg, i.p.) in acute drug challenges and tested for changes in wheel activity. Increased wheel activity produced by α-PVP or MDPV in the controls was attenuated in the α-PVP-KLH and MDPV-KLH vaccinated groups, respectively. Rectal temperature decreases produced by MDPV in the controls were reduced in duration in the MDPV-KLH vaccine group. A separate group (N = 19) was trained to intravenously self-administer α-PVP (0.05, 0.1 mg/kg/inf) and vaccinated with KLH or α-PVP-KLH, post-acquisition. Self-administration in α-PVP-KLH rats was initially higher than in the KLH rats but then significantly decreased following a final vaccine booster, unlike the stable intake of KLH rats. The data demonstrate that active vaccination provides functional protection against the effects of α-PVP and MDPV, in vivo, and recommend additional development of vaccines as potential therapeutics for mitigating the effects of designer cathinone derivatives.

Detectability of new psychoactive substances, 'legal highs', in CEDIA, EMIT, and KIMS immunochemical screening assays for drugs of abuse.

The increasing number of new psychoactive substances made available for recreational drug use has created a challenge for clinical toxicology and drug testing laboratories. As a consequence, the routine immunoassay drug testing may become less effective due to an increased occurrence of false negative and false positive screening results. This work aimed to extend the knowledge about analytical cross-reactivity of new substances in selected CEDIA, EMIT, and KIMS immunoassays for drugs-of-abuse screening. Urine standards were prepared by spiking blank urine with 45 new substances. Authentic urine samples from intoxication cases identified by liquid chromatography-tandem mass spectrometry (LC-MS/MS) were also studied. Several new psychoactive substances were demonstrated to display cross-reactivity in the immunoassays. CEDIA Amphetamine/Ecstasy and EMIT d.a.u. Amphetamine Class tests showed the highest reactivity towards the new drugs, which was expected since many have amphetamine-like structure and activity. In the samples from authentic cases, five new substances displayed 100% detection rate in the CEDIA Amphetamine/Ecstasy test. In conclusion, cross-reactivity data in routine urine drug screening immunoassays for a number of new psychoactive substances not studied before were reported. In both spiked and authentic urine samples, some new substances showed significant cross-reactivity and are thus detectable in the routine screening methods.

Immunochemical approach using monoclonal antibody against Δ(9)-tetrahydrocannabinolic acid (THCA) to discern cannabis plants and to investigate new drug candidates.

A monoclonal antibody (MAb-4A4) against Δ9- tetrahydrocannabinolic acid (THCA) showing extensive cross-reactivity against various cannabinoids was prepared. Using this antibody, a competitive enzyme-linked immunoassay (ELISA) was developed to detect Δ9-THCA in the range of 1 to 100 mg/ml. Various cannabinoids including Δ9-THC (Δ9-tetrahydrocannabinolic acid), Δ8-THCA (Δ8-tetrahydrocannabinolic acid), Δ8-THC (Δ8-tetrahydrocannabinol), CBD (cannabidiol), and CBN (cannabinol) were recognized by MAb-4A4, and their cross-reactivities were 55-1600% compared with Δ9-THCA (100%). This novel characteristic of this MAb enabled detection of marijuana residues in biological samples by detection of residual cannabinoids. The ELISA using MAb-4A4 was found to be applicable even for withered samples which contained only trace amounts of Δ9-THCA and Δ9-THC. In addition, this method using MAb-4A4 could be useful in forensic analysis since the MAb-4A4 also shows cross-reactivities against cannabinoid metabolites in body fluids. As well as forensic applications using this MAb, an investigation of new drug candidates focusing on cannabinoid metabolites arising from biotransformation in plant tissue was performed using immunochemical screening. The resulting new drug candidates were cannabinoid glycosides biotransformed by Pinellia ternata whose bioactivity is as yet unidentified. Our results indicate the utility of the application of ELISA using MAb-4A4 for further experiments involving marijuana and cannabinoids not only in the forensic field but also in the context of drug discovery.

Development and validation of a high-performance liquid chromatography method for the evaluation of niflumic acid cross-reactivity of two commercial immunoassays for cannabinoids in urine.

Niflumic acid is a nonsteroidal, anti-inflammatory drug widely prescribed in Greece. We recently noticed that this drug cross-reacts for cannabinoids in a kinetic interaction of microparticles in a solution (KIMS) immunoassay method but does not in an enzyme multiplied immunoassay technique (EMIT) immunoassay method. The objective of the study was to develop and validate a high-performance liquid chromatographic method in order to evaluate niflumic acid cross-reactivity in two commercial immunoassays for cannabinoids in urine, both in niflumic acid standards as well as in urine specimens obtained from subjects receiving niflumic acid. Urine niflumic acid standards were prepared in drug-free urine at 13 concentrations ranging from 1.25 to 1000 microg/mL. The standards gave presumptive positive cannabinoids results when analyzed by the KIMS immunoassay method when the concentration was above 2.5 microg/mL. None of the prepared standards gave a false-positive cannabinoid result when analyzed by the EMIT immunoassay method. By applying a 50 ng/mL cutoff for cannabinoids in these assays, all 55 urine specimens collected from the 5 subjects who participated gave negative results by the EMIT and false-positive results by the KIMS immunoassay method. It is concluded that KIMS is more prone to cross-reactions by niflumic acid compared to EMIT. Therefore, all positive screening tests for cannabinoids obtained by KIMS should be confirmed by another technique.

Psychotropic activity of the antialcohol preparation Proproten-100.

Antidepressant activity of Proproten-100 (antibodies to brain-specific S100 protein in ultralow doses) in patients with stage II alcohol dependence and alcohol withdrawal syndrome was studied in an open comparative clinical trial. The tricyclic antidepressant amitriptyline and benzodiazepine tranquilizer phenazepam served as reference preparations. Anxiolytic activity of Proproten-100 was highly competitive with that of phenazepam. Proproten-100 produced a stronger thymoleptic effect than amitriptyline. The preparation possessed activating properties, affected alcohol addiction, and did not cause side effects. Proproten-100 should undergo clinical tests during the therapy of neurotic, neurosis-like, and subdepressive borderline disorders.

[Impact of delta-9-tetrahydrocannabinol and its metabolites on the immune system]. / Impact du delta-9-tetrahydrocannabinol et ses métabolites sur le système immunitaire.

Many studies obvious impact of cannabinoids on the immune system. These studies follow the rapid advanced researches led in the immunology field. D9 Tetrahydrocannabinol and their metabolites decrease production of tumoral necrosis factor alpha. This decrease has for consequence a decrease of the apoptosis. Recent discovery of implication of cytokines in the phenomena of dependence, make the cannabis and their metabolites promoting agent induced dependence in association with drug abuse. The withdrawal of these products necessitates a intact immune system. D9 Tetrahydrocannabinnol and their metabolites inhibit production of IL-1 and gamma interferon. This inhibition has for consequence a decrease of 33% of the lymphocytes activity and an inhibition of 66% of the lymphocytes adenyl cyclase activity. The consumption of cannabis decreases immunological competence of macrophages, and alterate their essential role of trophicity of the nervous central system. Furthermore, inhibiting actions of cannabinoids on the cyclo-oxygenase, promote production of arachidonic acid degradation products. This compounds mimic the action of histamine, and inducing a raise of the vascular permeability and bronchospasm. These inolecules contributes at delayed reaction of anaphylaxia. However these actions of cannabinoids on the immune system promote their pull-back in cure of new pathology likes AIDS.

Antigenicity study of the new cognition-enhancing agent nefiracetam.

The potential antigenicity of the new cognition-enhancing agent nefiracetam (N-(2,6-dimethylphenyl)-2-(2-oxo-1-pyrrolidinyl)acetamide, DM-9384, CAS 77191-36-7) was investigated by tests for passive cutaneous anaphylaxis (PCA), systemic anaphylaxis (SA) and skin reaction in mice and guinea pigs. Mice were sensitized with nefiracetam (10-100 micrograms/animal) or nefiracetam-egg albumin (OA) mixture (10 micrograms/animal). No IgE antibodies to nefiracetam were detected in plasmas obtained from nefiracetam and nefiracetam-OA sensitized mice, indicating that nefiracetam has no immunogenicity or antigenicity eliciting potential. Guinea pigs were sensitized with nefiracetam (20-100 or 20 mg/kg) or nefiracetam-OA (2 mg/kg). No antibodies to nefiracetam were detected in the sera obtained from sensitized guinea pigs by PCA. Neither SA nor skin reaction was observed in the sensitized guinea pigs after the injection of challenge. These results suggest that nefiracetam possesses no antigenicity in mice and guinea pigs.

[Immunology and psychiatry]. / Immunologie et psychiatrie.

This review of scientific literature comes within the interest arisen from ten years by psychoneuroimmunology, a field connecting several disciplines and illustrating in a new way the psychosomatic relationships. A first category of works has been dedicated to the study of psychiatric disorders associated with various diseases concerning immunity (systemic diseases, endocrine diseases, cancers, infectious diseases), but also to the possible effect of distressing life events on the upset activation of immune functions, or even to the discovery of predisposing personality profiles (type C profile, depressive vulnerability). A second category of works concerns the analysis of the immune disturbances associated with certain psychiatric diseases, such as depression or schizophrenia, but also with some distressing life conditions, like bereavement. Animal experimentation and human experimentation provide various informations on the factors conditioning the immunomodulating effects of stress, sometimes in the direction of an inhibition, sometimes in the direction of an activation of immune functions. Finally, several papers shed light on the immunomodulating effects of psychotropic drugs. All these works open new horizons to the scientific knowledge and let us glimpse an extension to the use of psychologic therapeutics, as well of pharmacological ones as of non pharmacological.

[Hepatitis, cholestasis and amineptine (author's transl)]. / Hépatite, cholestase et amineptine.

Eight cases of hepatitis induced by amineptine chlorhydrate, a new tricyclic antidepressant are reported. Jaundice and/or biological changes occurred 16 to 75 days after the onset of treatment and with a total oral dose of 3,8 to 26 grams. Biologically, cholestasis and cytolysis have been observed simultaneously in 4 cases, cholestasis alone in 2 cases, and prevalent cytolysis in 2 others. After withdrawal of amineptine, clinical improvement was achieved within 10 to 21 days, whereas biological changes of hepatic function tests lasted for as long as 3 to 12 weeks. Physiopathology of such disorders is not clear : toxicity, enzyme induction, immunoallergy, may be considered as possible mechanisms.