A simple, quick and non-destructive approach for sampling drugs of abuse in tablets and blotter for qualitative analysis by paper spray mass spectrometry.

This study presents the development of a simple, fast, and inexpensive approach for the direct analysis of new psychoactive substances (NPS) in seized tablets and blotter paper, with improved sample preservation and increased analytical frequency. Paper triangles were gently rubbed against the surface of the samples containing synthetic drugs and then subjected to analysis by paper spray ionization mass spectrometry (PS-MS). Seized samples containing lysergic acid diethylamide (LSD) and several other substances from the classes of amphetamines, N-benzyl-substituted phenethylamines, synthetic cathinones, and synthetic cannabinoids, were analysed. Three types of paper were tested (filter paper, blotter paper, and synthetic paper) and several combinations of spray solvents were studied for the optimization. All samples were weighed and photographed before and after sequences of analysis in order to attest to the sample preservation. The results revealed that the approach is excellent for sample preservation, with less than 5% of mass loss even after 27 consecutive analyses. Moreover, no significant signal decreases were observed in mass spectrometry (MS) even after the experiments. It was possible to unequivocally identify illicit substances from seized samples (pills and blotter paper). By overcoming the solubilization and wet extraction process used for sample preparation, the waste was restricted to a volume of only 10 µL of solvent for the PS-MS analysis. The main advantage of our approach over existing methods is the sample preparation, which is simple and quick since the samples are just rubbed against the PS paper. This brings enormous benefits in terms of analytical frequency, economy of time and low consumption of solvents. Another important point is that the sample can remain intact for further analysis, which is crucial in forensic analysis.

Screening method for rapid classification of psychoactive substances in illicit tablets using mid infrared spectroscopy and PLS-DA.

Several new psychoactive substances (NPS) have reached the illegal drug market in recent years, and ecstasy-like tablets are one of the forms affected by this change. Cathinones and tryptamines have increasingly been found in ecstasy-like seized samples as well as other amphetamine type stimulants. A presumptive method for identifying different drugs in seized ecstasy tablets (n=92) using ATR-FTIR (attenuated total reflectance - Fourier transform infrared spectroscopy) and PLS-DA (partial least squares discriminant analysis) was developed. A hierarchical strategy of sequential modeling was performed with PLS-DA. The main model discriminated four classes: 5-MeO-MIPT, methylenedioxyamphetamines (MDMA and MDA), methamphetamine, and cathinones. Two submodels were built to identify drugs present in MDs and cathinones classes. Models were validated through the estimate of figures of merit. The average reliability rate (RLR) of the main model was 96.8% and accordance (ACC) was 100%. For the submodels, RLR and ACC were 100%. The reliability of the models was corroborated through their spectral interpretation. Thus, spectral assignments were performed by associating informative vectors of each specific modeled class to the respective drugs. The developed method is simple, fast, and can be applied to the forensic laboratory routine, leading to objective results reports useful for forensic scientists and law enforcement.

Genuine sildenafil tablets sold in Brazil disguised as MDMA.

MDMA and sildenafil are two examples among many substances consumed in "raves", as well as in other types of "recreative" social events nowadays. During the first six months of 2017, five cases of supposedly MDMA tablets seized by local law enforcement forces in the state of Minas Gerais, Brazil, and brought to our forensic laboratory for examination, attracted our attention among dozens of others, as the tablets apprehended in these cases were, in fact, colorfully painted versions of genuine, pentagon-shaped, sildenafil tablets, freely available for sale in local pharmacies and drugstores. Physical profiling, together with ATR-FTIR spectral matching, multi-component/deconvolution analysis and correlation were employed to prove that these tablets were genuine sildenafil tablets from a specific manufacturer, painted in a colorful way so that they could be marketed as MDMA tablets to unsuspecting buyers.

ATR-FTIR characterization of generic brand-named and counterfeit sildenafil- and tadalafil-based tablets found on the Brazilian market.

Viagra and Cialis are among the most counterfeited medicines in many parts of the world, including Brazil. Despite the many studies that have been made regarding discrimination between genuine and counterfeit samples, most published works do not contemplate generic and similar versions of these medicines and also do not explore excipients/adjuvants contributions when characterizing genuine and suspected samples. In this study, we present our findings in exploring ATR-FTIR spectral profiles for characterizing both genuine and questioned samples of several generic and brand-name sildenafil- and tadalafil-based tablets available on the Brazilian market, including Viagra and Cialis. Multi-component spectral matching (deconvolution), objective visual comparison and correlation tests were used during analysis. Besides from allowing simple and quick identification of counterfeits, results obtained evidenced the strong spectral similarities between generic and brand-named tablets employing the same active ingredient and the indistinguishability between samples produced by the same manufacturer, generic or not. For all sildenafil-based and some tadalafil-based tablets tested, differentiation between samples from different manufacturers, attributed to slight variations in excipients/adjuvants proportions, was achieved, thus allowing the possibility of tracing an unknown/unidentified tablet back to a specific manufacturer.

Brephedrone: A new psychoactive substance seized in Brazil.

At the beginning of 2015, sixty-two capsules containing red-brown crystals seized in a historical city in Brazil were sent to this forensic laboratory for drug testing analysis. The material was identified as being Brephedrone, a new psychoactive substance and a bromine synthetic cathinone that is related to serotonin transportation. This substance was analyzed by ATR-FTIR, GC-MS, LC-MS, 1H, 13C and 2D NMR. Brephedrone apprehensions have been previously reported in Finland, France and Spain. It was the first detection of this substance in the State of Minas Gerais. No reports or information regarding any other apprehension nor identification of Brephedrone in Brazil were known prior to the present case.

Johrei Effects on Water: A Pilot Study by Counting Drops.

BACKGROUND: Water is a key ingredient in the creation and sustainment of life. Moreover, water may be a key vehicle in the processes of energy healing, such as in the preparation of homeopathic remedies and spiritual treatments. Given these properties, the purpose of this study was to investigate whether the application of Johrei to water could lead to significant changes in the hydrodynamic behaviour of the fluid. METHODS: Four regular Johrei practitioners (P1, P2, P3 and P4) were selected for this study. Dripping water produced at the tip of a capillary was used as the hydrodynamic behaviour model. This behaviour was modelled mathematically, and tuning parameters φ4 and τ were used to assess significant differences in the dripping water samples that were subjected to Johrei compared with the samples that were not so treated. The tuning parameters were obtained using the Levenberg-Marquardt fitting algorithm. The data sets for each Johrei practitioner and the control experiment were analysed using ANOVA and a paired t-test. RESULTS: The mathematical model exhibited an excellent fit to our data, generating correlation coefficients (r) greater than or equal to 0.999. Significant differences were observed in both τ (P1 and P2, P < 0.05 and P < 0.01, respectively) and φ4 (P2, P < 0.01). As expected, no significant difference for the control experiment (without Johrei) was observed. CONCLUSIONS: Our results indicated a statistically significant change in the hydrodynamic behaviour of water correlated with Johrei treatment for 50% of the participating Johrei practitioners.

Rapid detection of NBOME's and other NPS on blotter papers by direct ATR-FTIR spectrometry.

Blotter paper is among the most common forms of consumption of new psychotropic substances (NPS), formerly referred as designer drugs. In many cases, users are misled to believe they are taking LSD when, in fact, they are taking newer and less known drugs like the NBOMEs or other substituted phenethylamines. We report our findings in quick testing of blotter papers for illicit substances like NBOMEs and other NPS by taking ATR-FTIR spectra directly from blotters seized on the streets, without any sample preparation. Both sides (front and back) of each blotter were tested. Collected data were analyzed by single- and multi-component spectral matching and submitted to chemometric discriminant analysis. Our results showed that, on 66.7% of the cases analyzed, seized blotters contained one or more types of NBOMEs, confirming the growing presence of this novel substances on the market. Matching IR signals were detected on both or just one side of the blotters and showed variable strength. Although no quantitative analysis was made, detection of these substances by the proposed approach serves as indication of variable and possibly higher dosages per blotter when compared to LSD, which showed to be below the detection limit of the applied method. Blotters containing a mescaline-like compound, later confirmed by GC-MS and LC-MS to be MAL (methallylescaline), a substance very similar to mescaline, were detected among the samples tested. Validity of direct ATR-FTIR testing was confirmed by checking the obtained results against independent GC-MS or LC-MS results for the same cases/samples.

Johrei, a Japanese healing technique, enhances the growth of sucrose crystals.

The effect of Johrei treatment on the crystallization of sucrose from supersaturated solutions was studied in comparison with the crystallization in untreated solutions. This work was performed assuming that Johrei enhances the natural mechanisms of equilibrium restoration in biological and nonbiological systems. The crystallization in Johrei-treated solutions as judged by statistical analysis was found to be faster than the crystallization in untreated solutions. A discussion is presented about the mechanisms possibly involved.

Living cell motility.

The motility of living eukaryotic cells is a complex process driven mainly by polymerization and depolymerization of actin filaments underneath the plasmatic membrane (actin cytoskeleton). However, the exact mechanisms through which cells are able to control and employ 'actin-generated' mechanical forces, in order to change shape and move in a well-organized and coordinated way, are not quite established. Here, we summarize the experimental results obtained by our research group during recent years in studying the motion of living cells, such as macrophages and erythrocytes. By using our recently developed defocusing microscopy technique, which allows quantitative analysis of membrane surface dynamics of living cells using a simple bright-field optical microscope, we were able to analyse morphological and dynamical parameters of membrane ruffles and small membrane fluctuations, study the process of phagocytosis and also measure values for cell refractive index, membrane bending modulus and cell viscosity. Although many questions still remain unanswered, our data seem to corroborate some aspects of recent physical models of cell membranes and motility.

Measuring optical and mechanical properties of a living cell with defocusing microscopy.

Defocusing microscopy (DM) is a recently developed technique that allows quantitative analysis of membrane surface dynamics of living cells using a simple bright-field optical microscope. According to DM, the contrast of defocused images is proportional to cell surface curvature. Although, until now, this technique was used mainly to determine size and amount of membrane shape fluctuations, such as ruffles and small random membrane fluctuations, in macrophages, its applications on cell biology extend beyond that. We show how DM can be used to measure optical and mechanical properties of a living macrophage, such as cell refractive index n, membrane bending modulus K(c), and effective cell viscosity eta for membrane-actin meshwork relaxation. Experimental data collected from defocused images of bone marrow-derived macrophages were used to evaluate these parameters. The obtained values, averaged over several different macrophages, are n = (1.384 +/- 0.015), K(c) approximately 3.2 x 10(-19) J, and eta approximately 459 Pa.s. We also estimate the amplitude of the small fluctuations to be of the order of 3 nm, which is around the step size of a polymerizing actin filament.

Homem XX: relato de três casos na faixa etária pediátrica / XX male: 3 case reports during childhood

São apresentados três pacientes com a condição clínica conhecida como "homem XX", rara na faixa etária pediátrica, caracterizada por um fenótipo masculino (em geral não associado a ambigüidade genital), testículos, porém cariótipo 46,XX. O diagnóstico costuma ser feito no adulto devido à esterilidade; na faixa etária pediátrica, ele é feito nos casos com ambigüidade genital ou ginecomastia. Na maioria dos pacientes é detectado o gene SRY (Sex-determining Region of the Y chromosome), o que explica a diferenciação testicular, porém em 20 por cento dos casos ele está ausente, o que torna evidente que a determinação gonadal é um processo dependente de múltiplos genes e fatores de transcrição. O diagnóstico de apenas 3 casos em dois serviços de referência num período de quase 30 anos indica sua raridade entre os casos de anomalias da diferenciação sexual.

Real-time measurements of membrane surface dynamics on macrophages and the phagocytosis of Leishmania parasites.

Defocusing microscopy was used for real-time observation and quantification of membrane surface dynamics in murine bone marrow macrophages. Small random membrane fluctuations (SRMF), possibly metabolic driven, were detected uniformly over all membrane surface. Morphological and dynamical parameters of ruffles, such as shape, dimensions, and velocity of propagation, were analyzed. Optical tweezers were used to promote phagocytosis of single Leishmania amazonensis amastigotes by selected macrophages. Analysis of ruffling activity on the macrophages before and during phagocytosis of the parasites indicated that increased ruffling response near forming phagosomes, most likely induced by the parasite, accelerates phagocytosis. The effects of temperature decrease on the dynamics of membrane surface fluctuations and on the phagocytosis of parasites were used to determine the overall activation energies involved in these processes. The values obtained support the existence of strong correlation between membrane motility and phagocytic capacity.

[XX male: 3 case reports during childhood]. / Homem XX: relato de três casos na faixa etária pediátrica.

We report on three patients with the clinical condition known as "XX male", which is uncommon in the pediatric age group. Patients have a male phenotype (usually without ambiguous genitalia) and testes; however, the karyotype is 46,XX. The diagnosis is usually made in adult life due to infertility; it may also be done by the pediatrician when there is ambiguous genitalia or gynecomastia. The SRY gene (Sex-determining Region of the Y chromosome) is detected in most cases, thus explaining the origin of testicular development; however, it is absent in 20% of the cases, thus indicating that gonadal determination is a complex process which depends on the interaction of many genes and transcription factors. The finding of only 3 cases in two reference services in a 30-year period indicates the rarity of this disorder among intersex cases.

Insuficiência renal crônica e hormônio de crescimento: uma associação com resultados promissores / Chronic renal failure and growth hormone

Crianças e adolescentes com insuficiência renal crônica (IRC) apresentam freqüentemente distúrbios do crescimento. A etiologia é multifatorial, incluindo baixa ingesta calórica, acidose, anemia, osteodistrofia renal e alterações na secreção dos fatores de crescimento. A anormalidade primária do eixo GH/IGF que causa a diminuição da velocidade de crescimento é a diminuição da secreção sérica de GH, que leva a uma diminuição da IGF-1 bioativa ou livre, associada a um aumento sérico da IGFBP-3, com mecanismo ainda não totalmente esclarecido. Apesar do mecanismo fisiopatológico do atraso de crescimento na IRC não estar totalmente elucidado, o uso de rhGH exógeno promove um efetivo aumento da velocidade de crescimento em crianças e adolescentes com IRC, tanto em tratamento dialítico quanto após o transplante renal. As alterações dos fatores de crescimento parecem ser predominantes na gênese da baixa estatura de pacientes com IRC, tendo em vista que a correção dos demais fatores etiológicos promove uma melhora mínima no crescimento, enquanto que o uso do rhGH causa uma marcada recuperação do crescimento em crianças com IRC e importante baixa estatura.

Avaliaçäo de um alimento com baixo teor de lactose em crianças portadoras de sindrome diarréica / Evaluation of a low lactose formula food in children with diahrreic syndrome

Estudaram-se 74 lactentes no primeiro ano de vida, de ambos os sexos, portadores de diarréia, verificando-se o desempenho de uma fórmula a base de proteínas do soro do leite, maltodextrina, gorduras e com baixo teor de lactose, suplementado com minerais e vitaminas - LLF-MILUPA. A idade média dos pacientes foi de 5,3 meses, sendo 46 portadores de diarréia aguda e 28 de diarréia aguda prolongada. As crianças foram divididas em dois grupos: Grupo I (/gI( - 36 realimentadas com LLF; Grupo II (GII) - 38 controles com leite de vaca. Foi elaborado um protocolo específico com o objetivo de colher dados relativos aos sintomas clínicos, avaliaçäo do estado nutricional e resultados de exames laboratoriais. As principais queixas foram: febre, vômitos, distensäo abdominal e cólicas. Quanto ao estado nutricional, verificaram-se 20 crianças eutróficas (10 GI e 10 GII), 22 com desnutriçäo protéico-calórica de I grau (10 GI e 12 GII), 24 de II grau (13 GI e 11 GII) e 8 de III grau (3 GI e 5 GII). A coprocultrua identificou bactérias enteropatogênicas em 15 casos (8 GI e 7 GII); a Giardia lamblia esteve presente em 6(4 GI e 2GII). O tempo médio de internaçäo, a tolerância e a eficácia foram semelhantes nos dois grupos estudados. Os resultados evidenciaram que a síndrome diarréica tem evoluçäo e resoluçäo satisfatórias com dieta habitual, desde que haja aporte hidroeletrolítico adequado. Em situaçöes como a intolerância a lactose, deve-se recorrer ao uso de fórmulas dietéticas especiais