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Lanaro R.,University of Campinas | Calemi D.B.A.,Criminalistic Institute of Sao Paulo | Togni L.R.,Criminalistic Institute of Sao Paulo | Costa J.L.,University of Campinas | And 3 more authors.
Journal of Psychoactive Drugs

Abstract: The ritualistic use of ayahuasca is becoming a global phenomenon. This beverage contains a combination of monoamine oxidase inhibitors (harmine, harmaline, and tetrahydroharmine) and N,N-dimethyltryptamine, the main substance responsible for its visionary effect. The recreational use of similar alkaloids and N,N-dimethyltryptamine has increased in recent years, mainly because of their hallucinogenic effects. In the present study, the concentrations of psychoactive alkaloids in three powder samples seized by the São Paulo State Police and nine ayahuasca aqueous extracts were analyzed by HPLC-DAD in an attempt to distinguish between illicit drugs and the religious beverage. The alkaloids detected (μg/mL) in the ayahuasca aqueous extracts were N,N-dimethyltryptamine (402–2070.3), harmaline (27.5–181.3), harmine (294.5–2893.8), and tetrahydroharmine (849.5–2052.5), whereas, of the three powder samples, one contained only N,N-dimethyltryptamine (82% and 2% w/w, respectively) while the other contained only harmaline (16%, w/w) and harmine (12%, w/w). The ritualistic use of ayahuasca involves oral intake and the probability of overdose is minimized by serotonergic stimulation of vagal pathways, leading to vomiting and diarrhea. In contrast, the recreational use of N,N-dimethyltryptamine involves consumption mainly by smoking or inhalation, both of which markedly increase its bioavailability and the potential for intoxications. © , Copyright © Taylor & Francis Group, LLC. Source

Togni L.R.,Criminalistic Institute of Sao Paulo | Lanaro R.,University of Campinas | Resende R.R.,Federal University of Minas Gerais | Costa J.L.,Criminalistic Institute of Sao Paulo | Costa J.L.,University of Campinas
Journal of Forensic Sciences

The content of ecstasy tablets has been changing over the years, and nowadays 3,4-methylenedioxymethamphetamine (MDMA) is not always present in the tablets. The aim of this study was to investigate the chemical composition in the seized tablets labeled as ecstasy. We analyzed samples from 150 different seizures made by Sao Paulo's State Police by gas chromatography-mass spectrometry. MDMA was present in 44.7% of the analyzed samples, and another twenty different active substances were identified in these tablets, such as caffeine, 2C-B, piperazines, amphetamines, phencyclidine, and others. Methamphetamine was present in 22% of these samples. The results demonstrate a huge shift in the pattern of trafficking of synthetic drugs, where MDMA has been replaced in tablets mostly by illicit psychoactive substances, in a clear attempt to bypass the law. The great variability in the tablets composition may lead to an increased risk of drug poisoning. © 2014 American Academy of Forensic Sciences. Source

Resende R.R.,Federal University of Minas Gerais | Resende R.R.,Institute of Learning and Research Santa Casa of BH IEPSC BH | Resende R.R.,Federal University of Sao Joao del Rei | Adhikari A.,Columbia University | And 6 more authors.
Biochimica et Biophysica Acta - Molecular Cell Research

Spontaneous Ca2+ events have been observed in diverse stem cell lines, including carcinoma and mesenchymal stem cells. Interestingly, during cell cycle progression, cells exhibit Ca2+ transients during the G1 to S transition, suggesting that these oscillations may play a role in cell cycle progression. We aimed to study the influence of promoting and blocking calcium oscillations in cell proliferation and cell cycle progression, both in neural progenitor and undifferentiated cells. We also identified which calcium stores are required for maintaining these oscillations. Both in neural progenitor and undifferentiated cells calcium oscillations were restricted to the G1/S transition, suggesting a role for these events in progression of the cell cycle. Maintenance of the oscillations required calcium influx only through inositol 1,4,5-triphosphate receptors (IP3Rs) and L-type channels in undifferentiated cells, while neural progenitor cells also utilized ryanodine-sensitive stores. Interestingly, promoting calcium oscillations through IP3R agonists increased both proliferation and levels of cell cycle regulators such as cyclins A and E. Conversely, blocking calcium events with IP3R antagonists had the opposite effect in both undifferentiated and neural progenitor cells. This suggests that calcium events created by IP3Rs may be involved in cell cycle progression and proliferation, possibly due to regulation of cyclin levels, both in undifferentiated cells and in neural progenitor cells. Source

Lanaro R.,University of Campinas | Costa J.L.,University of Campinas | Costa J.L.,Criminalistic Institute of Sao Paulo | Cazenave S.O.S.,Criminalistic Institute of Sao Paulo | And 4 more authors.
Journal of Forensic Sciences

In this work, two methods were developed to determine herbicides paraquat, glyphosate, and aminomethylphosphonic acid (AMPA) in marijuana samples by capillary electrophoresis. For paraquat analysis, sample was extracted with aqueous acetic acid solution and analyzed by capillary zone electrophoresis with direct UV detection. The running electrolyte was 50 mmol/L phosphate buffer (pH 2.50). For glyphosate and AMPA, indirect UV/VIS detection was used, as these substances do not present chromophoric groups. Samples were extracted with 5 mmol/L hydrochloric acid. The running electrolyte was 10 mmol/L gallic acid, 6 mmol/L TRIS, and 0.1 mmol/L CTAB (pH = 4.7). The methods presented good linearity, precision, accuracy, and recovery. Paraquat was detected in 12 samples (n = 130), ranging from 0.01 to 25.1 mg/g. Three samples were positive for glyphosate (0.15-0.75 mg/g), and one sample presented AMPA as well. Experimental studies are suggested to evaluate the risks of these concentrations to marijuana user. © 2014 American Academy of Forensic Sciences. Source

Resende R.R.,Federal University of Minas Gerais | Resende R.R.,Institute Ensino E Pesquisa Santa Casa Of Bh Iscm Bh | Da Costa J.L.,Criminalistic Institute of Sao Paulo | Da Costa J.L.,University of Sao Paulo | And 4 more authors.
Stem Cells and Development

Changes in intracellular Ca2+ concentration ([Ca 2+]i) play a central role in neuronal differentiation. However, Ca2+ signaling in this process remains poorly understood and it is unknown whether embryonic and adult stem cells share the same signaling pathways. To clarify this issue, neuronal differentiation was analyzed in two cell lines: embryonic P19 carcinoma stem cells (CSCs) and adult murine bone-marrow mesenchymal stem cells (MSC). We studied Ca2+ release from the endoplasmic reticulum via intracellular ryanodine-sensitive (RyR) and IP3-sensitive (IP3R) receptors. We observed that caffeine, a RyR agonist, induced a [Ca2+]i response that increased throughout neuronal differentiation. We also demonstrated a functional coupling between RyRs and L- but not with N-, P-, or Q-type Cav1 Ca 2+ channels, both in embryonal CSC and adult MSC. We also found that agonists of L-type channels and of RyRs increase neurogenesis and neuronal differentiation, while antagonists of these channels have the opposite effect. Thus, our data demonstrate that in both cell lines RyRs control internal Ca 2+ release following voltage-dependent Ca2+ entry via L-type Ca2+ channels. This study shows that both in embryonal CSC and adult MSC [Ca2+]i is controlled by a common pathway, indicating that coupling of L-type Ca2+ channels and RyRs may be a conserved mechanism necessary for neuronal differentiation. © Mary Ann Liebert, Inc. 2010. Source

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