Aplicação da Espectrometria de Massas Ambiente Por Paper Spray Ionization na Identificação e Quantificação de Cocaína e no Controle de Qualidade de Bebidas

Análises rápidas, baratas e eficientes são necessárias para as rotinas laboratoriais, principalmente quando aplicada em química forense. As utilizações de fontes ambientes permitem que as análises sejam diretas e rápidas. Assim, são opções inovadoras para melhorar a sensibilidade e seletividade. Neste trabalho, foi desenvolvida uma metodologia utilizando a espectrometria de massas ambiente por paper spray ionization mass spectrometry (PS-MS) para diferentes matrizes de dados: (i) identificar e quantificar cocaína e seus adulterantes a partir da cromatografia em camada delgada acoplada ao PS-MS; (ii) realizar a discriminação ou classificação de dez marcas brasileiras de cervejas em pilsen e lager; e (iii) identificar a falsificação controlada de Uisque com cachaça visando quantificar o volume de cachaça adicionado na mistura. Na análise de cocaína e adulterantes, os padrões foram identificados na CCD e analisados por PS-MS, e uma curva de calibração foi otimizada. A cerveja foi previamente fracionada usando a metodologia de extração em fase sólida (SPE) com água/metanol e os quatro extratos foram caracterizados no modo positivo de ionização, PS(+)MS. Para ambos os casos, foram aplicados de 20 µL do analito sobre a superfície do papel do PS-MS. Na quantificação de cachaça em misturas de whisk-cachaça. Os resultados indicam que, utilização da CCD acoplado ao PS-MS possibilita a identificação de cocaína e seus adulterantes uma ótima linearidade paras as curvas foi encontrada (cocaína (R2 = 0,9989), benzocaína, (R2 = 0,9934), a cafeína (R2 = 0,9988), a fenacetina (R2 = 0,9966), e a lidocaína (R2 = 0,9987)) e LOD inferior ao da técnica de CCD. Para as amostras de cerveja os sinais de açúcares se sobressaem sobre os demais grupos, e após a extração com água/metanol foi possível distinguir os espectros das diferentes marcas de cervejas e classifica-las em pilsen e lager pela técnica de PCA. Utilizando o PS(±)-MS e PLS com seleção de variáveis foi possível estimar quantitativamente o teor de cachaça misturado ao uísque. Com isso, utilização de fontes ambientes, como PS mostra-se eficaz para identificação de diversas matrizes, facilitando as análises de rotina

Morphological and physiological species-dependent characteristics of the rodent Grueneberg ganglion.

In the mouse, the Grueneberg ganglion (GG) is an olfactory subsystem implicated both in chemo- and thermo-sensing. It is specifically involved in the recognition of volatile danger cues such as alarm pheromones and structurally-related predator scents. No evidence for these GG sensory functions has been reported yet in other rodent species. In this study, we used a combination of histological and physiological techniques to verify the presence of a GG and investigate its function in the rat, hamster, and gerbil comparing with the mouse. By scanning electron microscopy (SEM) and transmitted electron microscopy (TEM), we found isolated or groups of large GG cells of different shapes that in spite of their gross anatomical similarities, display important structural differences between species. We performed a comparative and morphological study focusing on the conserved olfactory features of these cells. We found fine ciliary processes, mostly wrapped in ensheating glial cells, in variable number of clusters deeply invaginated in the neuronal soma. Interestingly, the glial wrapping, the amount of microtubules and their distribution in the ciliary processes were different between rodents. Using immunohistochemistry, we were able to detect the expression of known GG proteins, such as the membrane guanylyl cyclase G and the cyclic nucleotide-gated channel A3. Both the expression and the subcellular localization of these signaling proteins were found to be species-dependent. Calcium imaging experiments on acute tissue slice preparations from rodent GG demonstrated that the chemo- and thermo-evoked neuronal responses were different between species. Thus, GG neurons from mice and rats displayed both chemo- and thermo-sensing, while hamsters and gerbils showed profound differences in their sensitivities. We suggest that the integrative comparison between the structural morphologies, the sensory properties, and the ethological contexts supports species-dependent GG features prompted by the environmental pressure

Intrinsic aerobic capacity sets a divide for aging and longevity

<p><b>Rationale:</b> Low aerobic exercise capacity is a powerful predictor of premature morbidity and mortality for healthy adults as well as those with cardiovascular disease. For aged populations, poor performance on treadmill or extended walking tests indicates closer proximity to future health declines. Together, these findings suggest a fundamental connection between aerobic capacity and longevity.</p> <p><b>Objectives:</b> Through artificial selective breeding, we developed an animal model system to prospectively test the association between aerobic exercise capacity and survivability (aerobic hypothesis).</p> <p><b>Methods and Results:</b> Laboratory rats of widely diverse genetic backgrounds (N:NIH stock) were selectively bred for low or high intrinsic (inborn) treadmill running capacity. Cohorts of male and female rats from generations 14, 15, and 17 of selection were followed for survivability and assessed for age-related declines in cardiovascular fitness including maximal oxygen uptake (VO<sub>2max</sub>), myocardial function, endurance performance, and change in body mass. Median lifespan for low exercise capacity rats was 28% to 45% shorter than high capacity rats (hazard ratio, 0.06; P<0.001). VO<sub>2max</sub>, measured across adulthood was a reliable predictor of lifespan (P<0.001). During progression from adult to old age, left ventricular myocardial and cardiomyocyte morphology, contractility, and intracellular Ca<sup>2+</sup> handling in both systole and diastole, as well as mean blood pressure, were more compromised in rats bred for low aerobic capacity. Physical activity levels, energy expenditure (Vo<sub>2</sub>), and lean body mass were all better sustained with age in rats bred for high aerobic capacity.</p> <p><b>Conclusions:</b> These data obtained from a contrasting heterogeneous model system provide strong evidence that genetic segregation for aerobic exercise capacity can be linked with longevity and are useful for deeper mechanistic exploration of aging.</p&gt

When ring makes the difference: coordination properties of Cu2+/Cu+ complexes with sulfur-pendant polyazamacrocycles for radiopharmaceutical applications

Three polyazamacrocyclic ligands, i.e. 1,5,9-tris[2-(methylsulfanyl)ethyl]-1,5,9-triazacyclododecane (TACD3S), 1,4,7,10-tetrakis[2-(methylsulfanyl)ethyl]-1,4,7,10-tetrazacyclotridecane (TRI4S) and 1,4,8,11-tetrakis[2-(methylsulfanyl)ethyl]-1,4,8,11-tetrazacyclotetradecane (TE4S), were considered as potential chelators for the medically relevant copper radioisotopes. The ligands have been synthesized through facile, single-step reactions, and their acidity constants have been measured in aqueous solution at 25 degrees C. The kinetic, thermodynamic, electrochemical and structural properties of their Cu2+ and Cu+ complexes were investigated in aqueous solution at 25 degrees C using spectroscopic (UV-Visible, EPR, NMR) and electrochemical techniques (pH-potentiometric titrations, cyclic voltammetry and electrolysis). TACD3S was demonstrated to be unable to stabilize Cu2+, whereas for TRI4S and TE4S the formation of stable monocupric (CuL2+) and monocuprous (CuL+) complexes was detected. TRI4S coordinates Cu(2+)via a [4N] and a [4N]S array of donor atoms while with TE4S only the latter geometry exists. The thermodynamic stability and the kinetic inertness of the copper complexes formed by TACD3S, TRI4S and TE4S were compared with those previously reported for 1,4,7,10-tetrakis-[2-(methylsulfanyl)ethyl]-1,4,7,10-tetrazacyclododecane (DO4S) to unravel the influence of the ring size and the nitrogen donor array on the copper chelation properties of these sulfur-rich macrocycles. The copresence of four nitrogen atoms is an essential feature to allow effective copper coordination when a 12-member ring is employed, as the Cu2+-DO4S complexes were far more stable than those of Cu2+-TACD3S. Furthermore, the larger ring size of TRI4S and TE4S, when compared to DO4S, progressively increases the rate of the Cu2+ complexation reactions but decreases the thermodynamic stability of the Cu2+ complexes. Despite this, the ability of TRI4S and TE4S to stably accommodate both copper oxidation states makes them very attractive for application in nuclear medicine as they could avoid the demetallation after the biologically triggered Cu2+/Cu+ reduction

Caenorhabditis elegans provides an efficient drug screening platform for GNAO1-related disorders and highlights the potential role of caffeine in controlling dyskinesia

Dominant GNAO1 mutations cause an emerging group of childhood-onset neurological disorders characterized by developmental delay, intellectual disability, movement disorders, drug-resistant seizures and neurological deterioration. GNAO1 encodes the α-subunit of an inhibitory GTP/GDP-binding protein regulating ion channel activity and neurotransmitter release. The pathogenic mechanisms underlying GNAO1-related disorders remain largely elusive and there are no effective therapies. Here, we assessed the functional impact of two disease-causing variants associated with distinct clinical features, c.139A > G (p.S47G) and c.662C > A (p.A221D), using Caenorhabditis elegans as a model organism. The c.139A > G change was introduced into the orthologous position of the C. elegans gene via CRISPR/Cas9, whereas a knock-in strain carrying the p.A221D variant was already available. Like null mutants, homozygous knock-in animals showed increased egg laying and were hypersensitive to aldicarb, an inhibitor of acetylcholinesterase, suggesting excessive neurotransmitter release by different classes of motor neurons. Automated analysis of C. elegans locomotion indicated that goa-1 mutants move faster than control animals, with more frequent body bends and a higher reversal rate and display uncoordinated locomotion. Phenotypic profiling of heterozygous animals revealed a strong hypomorphic effect of both variants, with a partial dominant-negative activity for the p.A221D allele. Finally, caffeine was shown to rescue aberrant motor function in C. elegans harboring the goa-1 variants; this effect is mainly exerted through adenosine receptor antagonism. Overall, our findings establish a suitable platform for drug discovery, which may assist in accelerating the development of new therapies for this devastating condition, and highlight the potential role of caffeine in controlling GNAO1-related dyskinesia

Intravenous thrombolysis in the emergency department for the treatment of acute ischaemic stroke

BACKGROUND AND AIMS: Thrombolytic therapy with intravenous recombinant tissue plasminogen activator (rt-PA) improves outcome in patients with ischaemic stroke treated within 3 h of symptom onset, but its extended implementation is limited. A pilot study was designed to verify whether evaluation of patients with acute ischaemic stroke and their treatment with intravenous rt-PA in the emergency department (ED), followed by transportation to a semi-intensive stroke care unit, offers a safe and effective organisational solution to provide intravenous thrombolysis to acute stroke patients when a stroke unit (SU) is not available. METHODS: After checking for inclusion and exclusion criteria, ED doctors contacted the stroke team with a single page, located family members and urgently obtained computed tomography scan and laboratory tests. A stroke team investigator clinically assessed the patient, obtained written informed consent and supervised intravenous rt-PA in the ED. After treatment, the patient was transferred to the SU for rehabilitation and treatment of complications, under supervision of the same stroke team investigator. RESULTS: 52 patients were treated with intravenous rt-PA within 3 h of symptom onset. 20 patients (38%) improved neurologically after 24 h, the number increased to 30 (58%) after one week. At 3 months 22 patients had a favourable outcome (43%). The 3-month mortality rate was 12%. Symptomatic cerebral haemorrhage was observed in two patients (4%). CONCLUSIONS: Intravenous rt-PA administration in the ED is an effective organisational solution for acute ischaemic stroke when an SU is not established