Cannabinoids help to unravel etiological aspects in common and bring hope for the treatment of autism and epilepsy

Desde 1843 que as propriedades anticonvulsivantes da Cannabis são conhecidas pela ciência ocidental. Em 1980, ensaios clínicos demonstraram que canabidiol possui atividade antiepilética em pacientes de epilepsia refratária, sendo sonolência o único efeito colateral. O embargo imposto pela proibição do uso medicinal da Cannabis, no entanto, prejudicou imensamente o desenvolvimento científico e a exploração dessas propriedades. Multiplicam-se, contudo, os casos bem sucedidos de uso ilegal e sem orientação para o tratamento de síndromes caracterizadas por epilepsia e autismo regressivo. Os resultados corroboram evidências científicas que indicam a existência de processos etiológicos comuns entre o autismo e a epilepsia. Estudos em modelos animais confirmam envolvimento do sistema endocanabinoide. Esses avanços apontam o início de uma revolução no entendimento e tratamento desses transtornos.Since 1843 the anticonvulsant properties of Cannabis are known by the Western science. In 1980, clinical trials have shown that cannabidiol has antiepileptic activity in refractory epilepsy patients, with drowsiness as the only side effect. The embargo imposed by banning medicinal Cannabis use, however, harmed scientific development and the exploration of these properties. However, there is a growing number of successful cases of illegal use without guidance for the treatment of syndromes characterized by epilepsy and regressive autism. The results corroborate scientific evidence that indicates the existence of common etiological aspects between autism and epilepsy. Studies in animal models have confirmed involvement of the endocannabinoid system. These advances indicate the beginning of a revolution in the understanding and treatment of these disorders

Kenya: West Kenya postmarks

HiWATER: The Multi-Scale Observation Experiment on Evapotranspiration over heterogeneous land surfaces (MUSOEXE) Dataset - Flux Observation Matrix (stable isotopic observations

The time-depth of Corded Ware burial landscapes: A comparative study of Single Grave and Battle Axe burial alignments in Denmark, The Netherlands and Sweden

Barrow landscapes appeared in the third and second millennia BC throughout North-Western Europe; these first barrows were constructed by people of the Corded Ware culture and placed in alignments. This thesis is an interregional comparative study, to determine whether there is a pattern in the time-depths of the burial alignments of Trehuse-Sjørup-Dollerup in Denmark, Angelso-Emmerhout in The Netherlands and Lilla Beddinge in Sweden. The analysis is conducted by means of a literature study and the application of typochronologies. In recent research, the Corded Ware ‘culture’ is still seen as a widespread, unified social phenomenon that is the result of migration, but more emphasis is placed on the regional variability of this phenomenon. Regional variability is also what we see in the three case-studies; in fact, perhaps one may better speak of ‘local variability’, as each case-study reveals a remarkable variety even within one alignment. Even though there do seem to be interregionally shared traits, these are expressed in local practices. Despite the limitations of establishing a time-depth by means of typochronologies, all three alignments reveal a long use-life; even in the Bronze and Iron Ages, prehistoric people buried their dead here. Temporality seems to have been an important aspect of the ‘Corded Ware’ burial landscape

Collaborative mental health care : changing the landscape of mental health care?

In this thesis two major changes in the organization of mental health care delivery are explored. The first is collaborative mental health care in primary care, developed as an alternative way to treat common mental health disorders compared to the traditional referral and treatment practice. The collaborative care program followed the principles of stepped care. The first and least intensive treatment step was provided within the collaborative care program in the primary care setting. Treatment intensity was only stepped up through referral to specialized mental health care for patients who did not sufficiently respond to the first step. The traditional practice was direct referral and treatment within specialized care. In several studies we investigated effectiveness, short- and long-term efficiency, cost-effectiveness and whether the stepped care approach was appropriate for all patients instead of the matched care approach. The second change was the integration of eHealth in the collaborative care treatment model. In this study, implementation factors that could either inhibit or promote the uptake and utilization of blended collaborative care by mental health professionals and patients were assessed

Measuring Propargyl-Linked Drug Populations Inside Bacterial Cells, and Their Interaction with a Dihydrofolate Reductase Target, by Raman Microscopy

We report the first Raman spectroscopic study of propargyl-linked dihydrofolate reductase (DHFR) inhibitors being taken up by wild type Escherichia coli, Klebsiella pneumoniae, and Staphylococcus aureus cells. A novel protocol is developed where cells are exposed to the fermentation medium containing a known amount of an inhibitor. At a chosen time point, the cells are centrifuged and washed to remove the extracellular compound, then frozen and freeze–dried. Raman difference spectra of the freeze–dried cells (cells exposed to the drug minus cells alone) provide spectra of the compounds inside the cells, where peak intensities allow us to quantify the number of inhibitors within each cell. A time course for the propargyl-linked DHFR inhibitor UCP 1038 soaking into E. coli cells showed that penetration occurs very quickly and reaches a plateau after 10 min exposure to the inhibitor. After 10 min drug exposure, the populations of two inhibitors, UCP 1038 and UCP 1089, were ∼1.5 × 10⁶ molecules in each E. coli cell, ∼4.7 × 10⁵ molecules in each K. pneumonia cell, and ∼2.7 × 10⁶ in each S. aureus cell. This is the first in situ comparison of inhibitor population in Gram-negative and Gram-positive bacterial cells. The positions of the Raman peaks also reveal the protonation of diaminopyrimidine ring upon binding to DHFR inside cells. The spectroscopic signature of protonation was characterized by binding an inhibitor to a single crystal of DHFR

Carboxylation and Decarboxylation of Active Site Lys 84 Controls the Activity of OXA-24 β-Lactamase of Acinetobacter baumannii: Raman Crystallographic and Solution Evidence

The class D β-lactamases are characterized by the presence of a carboxylated lysine in the active site that participates in catalysis. Found in Acinetobacter baumannii, OXA-24 is a class D carbapenem hydrolyzing enzyme that exhibits resistance to most available β-lactamase inhibitors. In this study, the reaction between a 6-alkylidiene penam sulfone inhibitor, SA-1-204, in single crystals of OXA-24 is followed by Raman microscopy. Details of its reaction with SA-1-204 provide insight into the enzyme’s mode of action and help define the mechanism of inhibition. When the crystal is maintained in HEPES buffer, the reaction is fast, shorter than the time scale of the Raman experiment. However, when the crystal holding solution contains 28% PEG 2000, the reaction is slower and can be recorded by Raman microscopy in real time; the inhibitor’s Raman bands quickly disappear, transient features are seen due to an early intermediate, and, at approximately 2–11 min, new bands appear that are assigned to the late intermediate species. At about 50 min, bands due to all intermediates are replaced by Raman signals of the unreacted inhibitor. The new population remains unchanged indicating (i) that the OXA-24 is no longer active and (ii) that the decarboxylation of Lys84 occurred during the first reaction cycle. Using absorbance spectroscopy, a one-cycle reaction could be carried out in aqueous solution producing inactive OXA-24 as assayed by the chromogenic substrate nitrocefin. However, activity could be restored by reacting aqueous OXA-24 with a large excess of NaHCO₃, which recarboxylates Lys84. In contrast, the addition of NaHCO₃ was not successful in reactivating OXA-24 in the crystalline state; this is ascribed to the inability to create a concentration of NaHCO₃ in large excess over the OXA-24 that is present in the crystal. The finding that inhibitor compounds can inactivate a class D enzyme by promoting decarboxylation of an active site lysine suggests a novel function that could be exploited in inhibitor design

Measuring Drug-Induced Changes in Metabolite Populations of Live Bacteria: Real Time Analysis by Raman Spectroscopy

Raman difference spectroscopy is shown to provide a wealth of molecular detail on changes within bacterial cells caused by infusion of antibiotics or hydrogen peroxide. Escherichia coli strains paired with chloramphenicol, dihydrofolate reductase propargyl-based inhibitors, meropenem, or hydrogen peroxide provide details of the depletion of protein and nucleic acid populations in real time. Additionally, other reproducible Raman features appear and are attributed to changes in cell metabolite populations. An initial candidate for one of the metabolites involves population increases of citrate, an intermediate within the tricarboxyclic acid cycle. This is supported by the observation that a strain of E. coli without the ability to synthesize citrate, <i>gltA</i>, lacks an intense feature in the Raman difference spectrum that has been ascribed to citrate. The methodology for obtaining the Raman data involves infusing the drug into live cells, then washing, freezing, and finally lyophilizing the cells. The freeze-dried cells are then examined under a Raman microscope. The difference spectra [cells treated with drug] – [cells without treatment] are time-dependent and can yield population kinetics for intracellular species in vivo. There is a strong resemblance between the Raman difference spectra of E. coli cells treated with meropenem and those treated with hydrogen peroxide

Raman Spectra of Interchanging β‑Lactamase Inhibitor Intermediates on the Millisecond Time Scale

Rapid mix–rapid freeze is a powerful method to study the mechanisms of enzyme–substrate reactions in solution. Here we report a protocol that combines this method with normal (non-resonance) Raman microscopy to enable us to define molecular details of intermediates at early time points. With this combined method, SHV-1, a class A β-lactamase, and tazobactam, a commercially available β-lactamase inhibitor, were rapidly mixed on the millisecond time scale and then were flash-frozen by injection into an isopentane solution surrounded by liquid nitrogen. The “ice” was finally freeze-dried and characterized by Raman microscopy. We found that the reaction is almost complete in solution at 25 ms, giving rise to a major population composed of the <i>trans</i>-enamine intermediate. Between 25 and 500 ms, minor populations of protonated imine are detected that have previously been postulated to precede enamine intermediates. However, within 1 s, the imines are converted entirely to enamines. Interestingly, with this method, we can measure directly the turnover number of SHV-1 and tazobactam. The enzyme is completely inhibited at 1:4 ratio (enzyme:inhibitor) or greater, a number that agrees with the turnover number derived from steady-state kinetic methods. This application, employing non-intensity-enhanced Raman spectroscopy, provides a general and effective route to study the early events in enzyme–substrate reactions

HiWATER: Dataset of the spectral reflectance in the middle reaches of the Heihe River Basin

HiWATER: Dataset of the spectral reflectance in the middle reaches of the Heihe River Basi

HiWATER: Dataset of thermal infrared spectrum observed by BOMEM MR304 in the middle reaches of the Heihe River Basin

HiWATER: Dataset of thermal infrared spectrum observed by BOMEM MR304 in the middle reaches of the Heihe River Basi