Untargeted metabolomic study by liquid chromatography-mass spectrometry in brain tissues on the effects of combined cocaine and ethanol self-administration in male and female young rats.

The combined use of ethanol and cocaine is frequent among drug-abuse users and leads to further exacerbation of health consequences compared to individual consumption and this is of special concern during the transition to adulthood. Despite its high prevalence, the effect of combined consumption of cocaine and ethanol has been scarcely studied. In this work, we report the first untargeted metabolomic study in brain tissues to contribute to the advancement in the knowledge of the possible neurobiological effects of this polysubstance dependence. Liquid Chromatography coupled to high resolution Mass Spectrometry was employed to analyze three different brain tissues samples, prefrontal cortex, striatum and hippocampus, from male and female young rats exposed intravenously to a self-administration of these drugs. After optimizing the best sample treatment and selecting the chromatographic and detection conditions to find the maximum number of significant features (possible biomarker metabolites), the high resolution of the Orbitrap analyzer used in this work has made it possible to find up to 761 significant features with assigned molecular formula, of which up to 190 were tentatively identified and 44 unequivocally confirmed. The results demonstrated that the altered metabolic pathways are involved in multiple functions: receptor systems, such as the Glutamine-Glutamic acid-GABA axis or the catecholamine pathway, purinergic and pyrimidine pathways, fatty acids or oxidative stress, among others.

Untargeted metabolomic analysis to explore the impact of soil amendments in a non-conventional wastewater treatment.

As non-conventional wastewater treatment, vegetation filters make the most of the natural attenuation processes that occur in soil to remove contaminants, while providing several environmental benefits. However, this practice may introduce contaminants of emerging concern (CECs) and their transformation products (TPs) into the environment. A potential improvement to the system was tested using column experiments containing soil (S) and soil amended with woodchips (SW) or biochar (SB) irrigated with synthetic wastewater that included 11 selected CECs. This study evaluated: i) known CECs attenuation and ii) unknown metabolites formation. Known CECs attenuation was assessed by total mass balance by considering both water and soil media. An untargeted metabolomic strategy was developed to assess the formation of unknown metabolites and to identify them in water samples. The results indicated that SB enhanced CECs attenuation and led to the formation of fewer metabolites. Sorption and biodegradation processes were favored by the bigger surface area of particles in SB column, especially for compounds with negative charges. Incorporating woodchips into soil shortened retention times in the column, which reduced attenuation phenomena and resulted in the formation of significantly more metabolites. Incomplete biodegradation reactions, fostered by shorter retention times in SW column could mainly explain these results.

Metabolic fingerprinting of chemotherapy-resistant prostate cancer stem cells. An untargeted metabolomic approach by liquid chromatography-mass spectrometry.

Chemoresistance is one of the most important challenges in cancer therapy. The presence of cancer stem cells within the tumor may contribute to chemotherapy resistance since these cells express high levels of extrusion pumps and xenobiotic metabolizing enzymes that inactivate the therapeutic drug. Despite the recent advances in cancer cell metabolism adaptations, little is known about the metabolic adaptations of the cancer stem cells resistant to chemotherapy. In this study, we have undertaken an untargeted metabolomic analysis by liquid chromatography-high-resolution spectrometry combined with cytotoxicity assay, western blot, quantitative real-time polymerase chain reaction (qPCR), and fatty acid oxidation in a prostate cancer cell line resistant to the antiandrogen 2-hydroxiflutamide with features of cancer stem cells, compared to its parental androgen-sensitive cell line. Metabolic fingerprinting revealed 106 out of the 850 metabolites in ESI+ and 67 out of 446 in ESI- with significant differences between the sensitive and the resistant cell lines. Pathway analysis performed with the unequivocally identified metabolites, revealed changes in pathways involved in energy metabolism as well as posttranscriptional regulation. Validation by enzyme expression analysis indicated that the chemotherapy-resistant prostate cancer stem cells were metabolically dormant with decreased fatty acid oxidation, methionine metabolism and ADP-ribosylation. Our results shed light on the pathways underlying the entry of cancer cells into dormancy that might contribute to the mechanisms of drug resistance.

The Benefits of Self-Compassion in Mental Health Professionals: A Systematic Review of Empirical Research.

The importance of cultivating self-compassion is an often neglected issue among mental health professionals despite the risks to occupational well-being present in psychological care, such as burnout or compassion fatigue. In this context, this literature review has a twofold aim. Firstly, to contribute to raising awareness of the benefits of self-compassion among professionals, based on empirical research findings. Secondly, to coherently organize the available evidence on this topic, which to date appears scattered in a variety of articles. A systematic search on the APA PsycInfo database was conducted, and 24 empirical studies focused on the topic of the benefits of self-compassion in mental health professionals were finally selected. Concerning their methods, only 4 of the selected studies used experimental or quasi-experimental designs, 14 were cross-sectional studies, 3 presented qualitative research, and 3 were literature reviews. The research, regardless of methods used, points mainly to the benefits of self-compassion on the therapists' mental health and well-being; prevention of occupational stress, burnout, compassion fatigue, and secondary traumatization as well as improvement of therapeutic competencies and professional efficacy-related aspects. In the review, self-compassion appeared as a process that could explain the benefits (eg on burnout) of cultivating other skills (eg mindfulness). To further explore this point, an additional review included 17 studies focused on the effects of mindfulness or compassion-based interventions on therapists' self-compassion. In conclusion, our work joins those who have recommended the inclusion of self-compassion trainings in the curricula of mental health professionals.

Validity Evidence of the Spanish Version of the Mindful Attention Awareness Scale Using the Rasch Measurement Model.

The Mindful Attention Awareness Scale (MAAS) is the most cited instrument to measure dispositional mindfulness. However, some aspects of its validity are still under debate. We aimed to assess different sources of validity evidence (i.e., response processes, content, internal structure, reliability, and relations with external variables) of the MAAS scores in a sample of Spanish-speaking participants (N = 812) applying Rasch modeling. The items formed an essentially unidimensional structure, the item hierarchy was similar to that of previous comparable studies, the items were well targeted, and the ordering of persons along the construct was adequate. Moreover, measures were invariant across four age groups and three groups based on meditation practice, and correlated as expected with a variety of well-being variables. In sum, our findings supported the interpretation of MAAS scores as a measure of mindfulness in our sample of Spanish-speaking participants. Any other specific inference should be tested.

Reductions in experiential avoidance explain changes in anxiety, depression and well-being after a mindfulness and self-compassion (MSC) training.

OBJECTIVE: The mindfulness and self-compassion (MSC) protocol has shown efficacy in reducing mental illness symptoms and increasing well-being. However, little is known on how the positive outcomes are produced. This study explores how reductions in experiential avoidance following MSC training may explain changes in the participants' levels of anxiety, depression, and well-being. METHODS: The 8-week protocol-based MSC training was delivered to 50 participants, and pre- to post- intervention changes in anxiety, depression, and well-being were measured. A series of mediation models were conducted, with changes in self-compassion as predictor, changes in experiential avoidance as mediator, and changes in mental health and well-being as outcome variables. Point estimates and bootstrap-corrected 95% confidence intervals were calculated to analyse indirect effects through experiential avoidance, by means of structural equation modeling (SEM). RESULTS: Following MSC training, participants increased their levels of self-compassion, reduced experiential avoidance, and enhanced mental health (i.e., anxiety and depression symptoms) and well-being scores. Increases in self-compassion were associated with decreases in experiential avoidance, which in turn were connected with changes in anxiety, depression, and well-being from pre- to post-training. The indirect path through changes in experiential avoidance represented moderate to large proportions of the total effects of self-compassion change-scores on anxiety, depression, and well-being change-scores. CONCLUSIONS: Reducing experiential avoidance and increasing psychological flexibility may be a key effect of MSC training linked to improvements of the participants' mental health and well-being scores. Self-compassion practices could exert effects on anxiety, depression and well-being mainly through promoting reductions in experiential avoidance.

Relationships between Mindfulness, Purpose in Life, Happiness, Anxiety, and Depression: Testing a Mediation Model in a Sample of Women.

Mindfulness is connected to positive outcomes related to mental health and well-being. However, the psychological mechanisms that account for these relationships are largely unknown. A multiple-step multiple mediator structural equation modeling (SEM) model was tested with mindfulness as the independent variable; purpose in life and behavioral activation as serial mediators; and happiness, anxiety, and depression as outcome measures. Data were obtained from 1267 women. Higher mindfulness was associated with higher levels of happiness and lower anxiety and depression symptoms. The association of mindfulness with the outcome variables could be partially accounted for by purpose in life and behavioral activation. The SEM model explained large proportions of variance in happiness (50%), anxiety (34%), and depression (44%) symptoms. Mindfulness is associated with both a sense of purpose in life and engagement in activities, which are also connected with positive outcomes. Moreover, having purposes in life is linked to higher levels of behavioral activation.

What Do Meta-Analyses Have to Say About the Efficacy of Neurofeedback Applied to Children With ADHD? Review of Previous Meta-Analyses and a New Meta-Analysis.

Objective: We reviewed previously published meta-analyses of neurofeedback applied to children with ADHD and conducted a new meta-analysis of randomized controlled trials (RCTs) that updates previous results and incorporates methodological modifications. Method: Searches were carried out through PubMed, Sage, PsycINFO, SpringerLink, and Psicodoc. We used Hedges' (adjusted) g and a random-effects model. To assess heterogeneity, Q and I2 were calculated. We performed different analyses depending on the control groups, ADHD symptoms, pre- and/or posttreament data used and symptom evaluator. Results: We reviewed seven meta-analyses, and 17 studies were incorporated into the meta-analysis. RCTs support the efficacy of neurofeedback applied to ADHD when most proximal evaluators assess symptoms. Neurofeedback significantly improves inattention symptoms when possibly blinded evaluators assess symptoms. The preliminary results suggest that stimulant medication is more effective than neurofeedback. Conclusion: New RCTs that establish links between ADHD symptom measurements, subjects' learning after neurofeedback, and neurophysiological measures could improve the quality of the conclusions.

The effects of combined intravenous cocaine and ethanol self-administration on the behavioral and amino acid profile of young adult rats.

Under paradigms of combined intravenous cocaine and ethanol self-administration, the effects on behavior have been poorly explored. Numerous studies have found sex differences in amino acids profile and behavioral responses to each drug, yet few have focused on the interactions between cocaine and ethanol. The main objective of this work was to explore the acquisition and maintenance of intravenous self-administration behavior with a combination of cocaine and ethanol in male and female young adult rats. Likewise, the amino acids profile in blood plasma was quantified 48 hours after the last self-administration session. Male and female 52 days old Wistar rats were randomly assigned to one of 3 groups: i) saline control, ii) cocaine (1 mg/kg bodyweight/injection) and iii) cocaine and ethanol (1 mg + 133 mg/kg bodyweight/ injection). After 24 self-administration sessions carried out on a fixed-ratio-1 schedule, with a limit of 15 doses per session, 14 plasma amino acids were quantified by mean Capillary Electrophoresis technique. The curve of cocaine and ethanol combined self-administration was similar to that associated with cocaine administration alone, with females acquiring self-administration criterion before males. The self-administration of cocaine and ethanol altered the plasma concentration and relative ratios of the amino acid L-Tyrosine. In our intravenous self-administration model, females appeared more vulnerable to acquire abusive consumption of the cocaine and ethanol combination, which altered plasma L-Tyrosine levels.

Self-compassion, meaning in life, and experiential avoidance explain the relationship between meditation and positive mental health outcomes.

OBJECTIVE: Despite consistent evidence for the beneficial effects of meditation on mental health, little is known about the mechanisms that make mindfulness meditation effective. METHOD: The levels of mental health, self-compassion, presence of meaning in life, and experiential avoidance of meditators (n = 414) and nonmeditators (n = 414) were measured and compared. Bootstrap-based structural equation modeling (SEM) modeling analyses were used to test multiple-step multiple-mediator models. RESULTS: Meditation was positively associated with mental health, although the regularity of practice was an influential element to be considered. Significant indirect effects of meditation on mental health through self-compassion, meaning in life, and experiential avoidance were found. SEM models were able to account for 58% of the variance in mental health scores. CONCLUSIONS: Self-compassion, presence of meaning in life, and reduced experiential avoidance may be active components of healthy meditation practices. Identifying the mechanisms involved in effective meditation practices has relevant implications for well-being and mental health-promoting interventions.

Automated open-access liquid chromatography high resolution mass spectrometry to support drug discovery projects.

The need of a continuous productivity increases in medicinal chemistry laboratories of the pharmaceutical industry motivated the development, over the years, of new software solutions to enable Open-Access in many analytical techniques such as NMR or LC, among others, to characterize and assess the purity of new molecules. These approaches have been widely spread in LC with low resolution MS systems, but similar automated platforms have been rather less explored with high resolution MS. In this work, an improved Automated Open-Access methodology on an UHPLC with DAD coupled to ESI and quadrupole time-of-flight MS system is described. Detailed reports from standard UHPLC-MS runs containing chromatograms and different spectra (MS with different fragmentation) are automatically sent to the chemists. High resolution MS data is typically achieved within ± 1 mDa mass accuracy regardless of sample concentration. Upon training, chemists log-in samples into the system by selecting appropriate methods, being able to interpret the results by themselves in 95% of the cases. The instrument is working unattended, except for a limited number of samples (5%) which require more complex experiments. To the best of our knowledge, this is the first time a completely automated Open-Access LC-HRMS approach has been implemented for medicinal chemists of a pharmaceutical industry.

Sheathless CE-MS based metabolic profiling of kidney tissue section samples from a mouse model of Polycystic Kidney Disease.

Capillary electrophoresis-mass spectrometry (CE-MS) using a sheathless porous tip interface emerged as an attractive tool in metabolomics thanks to its numerous advantages. One of the main advantages compared to the classical co-axial sheath liquid interface is the increased sensitivity, while maintaining the inherent properties of CE, such as a high separation efficiency and low sample consumption. Specially, the ability to perform nanoliter-based injections from only a few microliters of material in the sample vial makes sheathless CE-MS a well-suited and unique approach for highly sensitive metabolic profiling of limited sample amounts. Therefore, in this work, we demonstrate the utility of sheathless CE-MS for metabolic profiling of biomass-restricted samples, namely for 20 µm-thick tissue sections of kidney from a mouse model of polycystic kidney disease (PKD). The extraction method was designed in such a way to keep a minimum sample-volume in the injection vial, thereby still allowing multiple nanoliter injections for repeatability studies. The developed strategy enabled to differentiate between different stages of PKD and as well changes in a variety of different metabolites could be annotated over experimental groups. These metabolites include carnitine, glutamine, creatine, betaine and creatinine. Overall, this study shows the utility of sheathless CE-MS for biomass-limited metabolomics studies.

Recent contributions for improving sensitivity in chiral CE.

The flexibility and versatility of the chiral CE are unrivaled and the same instrumentation can be used to separate a diverse range of analytes, both large and small molecules, whether charged or uncharged. However, one of the disadvantages is generally thought to be the poor sensitivity of ultraviolet (UV) detection, which is the most popular among CE detectors. This review focuses on methodologies and applications regarding improvements of sensitivity in chiral CE published in the last 2 years (June 2015 until May 2017). This contribution continues to update this series of biannual reviews, first published in Electrophoresis in 2006. The main body of the review brings a survey of publications organized according to different approaches to detect a low amount of analytes, either by sample treatment procedures or by in-capillary sample preconcentration techniques, both using UV detection, or even by employing detection systems more sensitive than UV absorption, such as LIF or MS. This review provides comprehensive tables listing the new approaches in sensitive chiral CE with categorizing by the fundamental mechanism to enhance the sensitivity, which provide relevant information on the strategies employed. The concluding remarks in the final part of the review evaluate present state of art and the trends for sensitivity enhancement in chiral CE.

The role of cognitions in short-term temporal changes in dental fear among Australian adults.

OBJECTIVES: To determine if temporal changes in dental fear over a 4-month period are predicted by changed cognitive vulnerability-related perceptions of going to the dentist. METHODS: Australian adults (n = 484) completed mailed questionnaires at baseline and follow-up, containing measures of dental fear, cognitive vulnerability-related perceptions, dental services received, and possible aversive experiences during the study period. Change in dental fear was the main outcome measure, categorized as decreased (Fear- ), unchanged (Fear0 ), and increased (Fear+ ). RESULTS: Across the study period, 15.5 percent of people had Fear- , 73.4 percent had Fear0 , and 11.1 percent had Fear+ . In a multinomial logistic regression, after controlling for participant gender, income, time since last dental visit at baseline, dental fear at baseline and experiencing an aversive event, increased vulnerability-related perceptions were significantly associated with Fear+ (OR = 2.83, P < 0.001) while decreased vulnerability-related perceptions were associated with Fear- (OR = 0.17, P < 0.001). CONCLUSIONS: This study found, across a relatively short 4-month period, that increased vulnerability-related perceptions of visiting the dentist predicted increased dental fear while decreased vulnerability-related perceptions predicted decreased dental fear. More appropriate study designs, such as longitudinal designs, and longer follow-up periods are needed to determine the possible causal nature of these associations.

Evaluación de una intervención para la mejora de las competencias de regulación interpersonal del afecto y el bienestar laboral en una muestra de operadores penitenciarios del Uruguay / Evaluation of a Training Program aiming to Improve Interpersonal Emotion Regulation Strategies and Occupational Wellbeing in a Uruguayan Prison

Resumen Según estudios previos, el uso de estrategias controladas para la regulación interpersonal (ECRI) del afecto de otros orientadas a su mejora incrementa el bienestar de quien las emplea, mientras el uso de ECRI para empeorar el afecto lo deteriora. El consumo y recuperación de recursos de autoregulación de cada ECRI explican estos resultados. Mediante un diseño cuasiexperimental se evalúa la eficacia de la formación en ECRI para mejorar el bienestar en operadores de prisiones. Los resultados reflejan mayor bienestar y menor uso de ECRI de empeoramiento tras la formación (N= 21), al comparar con el grupo control (N= 18). No hubo diferencias para las ECRI de mejora. Los cambios en las ECRI de empeoramiento predicen negativamente cambios en el bienestar.

Abstract Previous research show a positive relationship between using IARS to improve other people's affect and the level of wellbeing of the agent implementing them, whereas using IARS to worsen other's affect can worsen the agent's wellbeing. These results can be explained as the balance between the self-control resources drained and recovered by each IARS. We evaluate the efficacy of a training program to increase prison officers' wellbeing and focused on IARS. Results, based on a quasi-experimental design, revealed a higher level of wellbeing and a lower increased on worsening strategies among the training participants (N=21), compared with a control group (N=18). Results were not significant for improving strategies. Changes in the use of IARS negatively predicted changes in wellbeing.

Investigation on the combined effect of cocaine and ethanol administration through a liquid chromatography-mass spectrometry metabolomics approach.

Alcohol is the most widely consumed legal drug, whereas cocaine is the illicit psychostimulant most commonly used in Europe. The combined use of alcohol and cocaine is frequent among drug-abuse consumers and leads to further exacerbation of health consequences compared to individual consumption. The pharmacokinetic and metabolic interactions leading to an increase in their combined toxicity still remains poorly understood. Here, the first metabolomics study of combined cocaine and ethanol chronic exposure effects is reported. A Liquid Chromatography strategy based on sample derivatization with 9-fluorenylmethyloxycarbonyl chloride and using a C18 column coupled to high resolution Mass Spectrometry (time of flight analyzer) was employed to analyze plasma from rats exposed intravenously to these drugs in a 52-min analysis. Using a combination of non-supervised and supervised multivariate analysis the metabolic differences between our experimental groups were explored and unraveled. A comparative analysis of the individual models and their variable importance in the projection values have shown that every experiment intervention includes a subset of specific metabolites. Eleven of these metabolites were annotated, where eight were unequivocally identified using standards and three were tentatively identified by matching the MS/MS spectra to libraries. The results demonstrated that the affected metabolic pathways were mainly those related to the metabolism of different amino acids.

A novel method for the quality control of saffron through the simultaneous analysis of authenticity and adulteration markers by liquid chromatography-(quadrupole-time of flight)-mass spectrometry.

A liquid chromatography-(quadrupole-time of flight)-mass spectrometry methodology was developed to assess the authenticity of saffron through the analysis of a group of kaempferol derivatives recently proposed as novel authenticity markers as a result of a metabolomic study of saffron (kaempferol 3-O-glucoside, kaempferol 3-O-sophoroside, kaempferol 3,7-O-diglucoside, kaempferol 3,7,4'-O-triglucoside, kaempferol 3-O-sophoroside-7-O-glucoside). Geniposide was also studied as an adulteration marker of saffron with gardenia. The optimized chromatographic conditions enabling the simultaneous separation of glycosylated kaempferols and geniposide consisted of the use of a C18 column and an elution gradient with acetonitrile and water as mobile phases (both with formic acid at 0.1%). A strategy was proposed to evaluate the minimum quantifiable adulteration percentage which was established at a 0.2% regardless of the adulterant employed. The analysis of nineteen commercial samples showed the method to be specific and suitable for saffron quality control.

Design of strategies to study the metabolic profile of highly polar compounds in plasma by reversed-phase liquid chromatography-high resolution mass spectrometry.

Amino acids and related compounds are paramount analytes which are involved in numerous metabolic pathways. Most of these compounds are unable to be retained on Liquid Chromatography with Reversed-Phase stationary phases due to their high hydrophilic character. An interesting strategy is to reduce their polarity through their derivatization with a labelling reagent, such as the commercially available 9-fluorenylmethyloxycarbonyl (FMOC) which forms stable complexes with primary and secondary amine moieties rapidly. Although some derivatization reagents have been employed in the study of metabolic profiles, as far as we know, FMOC has never been employed for this purpose. In this work, it is demonstrated that the use of RP-LCMS(TOF) using a C18 column and FMOC as labelling agent enables the determination of a larger number of hydrophilic compounds (proteinogenic amino acids, non-proteinogenic amino acids, and biogenic amines) when compared to the use of a fully-wettable pentafluorophenyl column in fully-aqueous conditions (gradient starting in 0% of organic solvent) and HILIC column, both without using compound derivatization. Different strategies for plasma protein elimination were also carefully evaluated. Results revealed that ultrafiltration (UF) offered a lower variability from sample to sample when compared to the protein precipitation (PP) method (from 2 to 12 times lower variability found in UF). Additionally, UF preserved a larger number of possible compounds when compared to the PP approach: 4631 unique molecular features with UF, 666 unique molecular features with PP.

Chiral separation of a basic drug with two chiral centers by electrokinetic chromatography for its pharmaceutical development.

A chiral method using capillary electrophoresis was developed for the separation of the four stereoisomers of a new chiral substance currently undergoing drug development as single enantiomer. After the selection of highly sulfated ß-CD as chiral selector, an exhaustive study on the influence of several experimental variables on the resolution was performed, being the substitution degree of the CD a very decisive factor. Run time and resolutions were about 20min and higher than 2.0, respectively. The method was validated in terms of selectivity, linearity, accuracy, precision, and limits of detection and quantitation according to the requirements of the International Conference on Harmonisation for the determination of the chiral purity of a drug substance. The usefulness of the method was demonstrated in the control of stereoisomeric impurities in raw material as well as in the determination of the chiral stability of the drug in the solid state and in dosage forms used in safety assessment. Finally, the chiral method was used to investigate the possible in vivo inversion in biological samples.

Enantioseparation of the constituents involved in the phenylalanine-tyrosine metabolic pathway by capillary electrophoresis tandem mass spectrometry.

Catecholamines dopamine, norepinephrine, and epinephrine are well-known neurotransmitters playing different roles in the nervous and endocrine system. These compounds are biologically synthesized in the phenylalanine-tyrosine pathway which consists on the successive conversion of l-phenylalanine into l-tyrosine, l-3,4-dihydroxyphenylalanine (L-DOPA), dopamine, norepinephrine, and epinephrine. This work describes the development of an enantioselective CE-ESI-MS2 methodology enabling, for the first time, the simultaneous enantioseparation of all the constituents involved in the Phe-Tyr metabolic pathway, since all these compounds except dopamine are chiral. The developed method was based on the use of a dual CDs system formed by 180mM of methyl-ß-CD and 40mM of 2-hydroxypropyl-ß-CD dissolved in 2M formic acid (pH 1.2) and presented the advantage of avoiding the use of any time-consuming labelling procedure. LODs ranged from 40 to 150nM and the unequivocal identification of the compounds investigated was achieved through their MS2 spectra. The applicability of this methodology to the analysis of biological samples (rat plasma) was also demonstrated.