Experimental and Computational Insights into the Overall Water Splitting Reaction by the Fe-Co-Ni-P Electrocatalyst.

Nonprecious transition-metal phosphides (TMPs) are versatile materials with tunable electronic and structural properties that could be promising as catalysts for energy conversion applications. Despite the facts, TMPs are not explored thoroughly to understand the chemistry behind their rich catalytic properties for the water splitting reaction. Herein, spiky ball-shaped monodispersed TMP nanoparticles composed of Fe, Co, and Ni are developed and used as efficient electrocatalysts for hydrogen and oxygen evolution reaction (HER, OER), and overall water splitting in alkaline medium; and their surface chemistry was explored to understand the reaction mechanism. The optimized Fe0.5CoNi0.5P catalyst shows attractive activities of HER and OER with low overpotentials and Tafel slopes, and with high mass activities, turnover frequencies, and exchange current densities. When applied to overall water splitting, the electrolyzer Fe0.5CoNi0.5P||Fe0.5CoNi0.5P cell can reach a 10 mA cm-2 current density at cell voltages of only 1.52 and 1.56 V in 1.0 M and 30 wt % KOH, respectively, much lower than those of commercial IrO2||Pt/C. The optimized electrolyzer with sizable numbers of chemically active sites exhibits superior durability up to 70 h and 5000 cycles in 1.0 M KOH and can attain a current density as high as 1000 mA cm-2, showing a class of efficient bifunctional electrocatalysis. Experimental and density functional theory-based mechanistic analyses reveal that surface reconstruction takes place in the presence of KOH to form the TMP precatalyst, which results in high coverage of oxygen active species for the OER with a low apparent activation energy (Ea) for conversion of *OOH to O2. These also evidenced the thermoneutral adsorption of H* for the efficient HER half-reaction.

A New Lignan from Annona squamosa L. (Annonaceae) Demonstrates Vasorelaxant Effects In Vitro.

Esquamosan, a new furofuran lignan, has been isolated by bio-guided assays from the methanolic extract of the leaves of Annona squamosa L., and its structure was elucidated by spectroscopic methods. Esquamosan inhibited the rat aortic ring contraction evoked by phenylephrine in a concentration-dependent manner and showed an inhibitory effect on vasocontraction of the depolarized aorta with high-concentration potassium. The vasorelaxant effect by esquamosan could be attributed mainly to the inhibition of calcium influx from extracellular space through voltage-dependent calcium channels or receptor-operated Ca2+ channels and also partly mediated through the increased release of NO from endothelial cells. The ability of esquamosan to modify the vascular reactivity of rat aortic rings incubated with high glucose (D-glucose 55 mM) was then evaluated, and this furofuran lignan reverted the endothelium-dependent impairment effect of high glucose in rat aortic rings. The antioxidant capacity of esquamosan was assessed using DPPH and FRAP assays. Esquamosan exhibited a similar antioxidant capacity compared to ascorbic acid, which was used as a positive control. In conclusion, this lignan showed a vasorelaxant effect, free radical scavenging capacity, and potential reductive power, suggesting its potential beneficial use to treat complex cardiometabolic diseases due to free radical-mediated diseases and its calcium antagonist effect.

Microproteins: Overlooked regulators of physiology and disease.

Ongoing efforts to generate a complete and accurate annotation of the genome have revealed a significant blind spot for small proteins (<100 amino acids) originating from short open reading frames (sORFs). The recent discovery of numerous sORF-encoded proteins, termed microproteins, that play diverse roles in critical cellular processes has ignited the field of microprotein biology. Large-scale efforts are currently underway to identify sORF-encoded microproteins in diverse cell-types and tissues and specialized methods and tools have been developed to aid in their discovery, validation, and functional characterization. Microproteins that have been identified thus far play important roles in fundamental processes including ion transport, oxidative phosphorylation, and stress signaling. In this review, we discuss the optimized tools available for microprotein discovery and validation, summarize the biological functions of numerous microproteins, outline the promise for developing microproteins as therapeutic targets, and look forward to the future of the field of microprotein biology.

Biorefinery and sustainability for the production of biofuels and value-added products: A trends analysis based on network and patent analysis.

Biorefineries are modern mechanisms used for producing value-added products and biofuels from different biomass sources. However, a crucial challenge is to achieve a sustainable model for their adequate implementation. Challenges related to technical efficiency and economic feasibility are two of the most relevant problems. Therefore, the present study sought to determine the current trends in basic research and technological development around biorefining and sustainability. We carried out a co-occurrence analysis and a patent analysis using data obtained from the Scopus and Lens databases to provide a general overview of the current state of this area of knowledge. The co-occurrence analysis intends to provide an overview of biorefining and sustainability based on terms associated with these two concepts as a starting point to determine the progress and existing challenges of the field. The results of the patent analysis consisted in identifying the main technological sectors, applicants, and territories where inventions associated with biorefining are registered. The analysis of the information showed that bioeconomy, techno-economic aspects, circular economy, technical issues associated with biomass production, and biofuels represent the focal point of basic research in a wide range of disciplines. Technology development is focused on fermentation, enzymes, and microorganisms, among other areas, which shows the validity of these traditional techniques in addressing the problems faced by the bioeconomy. This scenario shows that developed economies are the driving force behind this area of knowledge and that the PCT system is fundamental for the protection and commercialization of these inventions in places different from where they originated. Furthermore, the challenge lies in learning to work in alternative and complementary technological sectors, beyond microbiology and enzyme applications, in pursuit of the sector's technical and economic feasibility.

Dwarf Open Reading Frame (DWORF) Gene Therapy Ameliorated Duchenne Muscular Dystrophy Cardiomyopathy in Aged mdx Mice.

Background Cardiomyopathy is a leading health threat in Duchenne muscular dystrophy (DMD). Cytosolic calcium upregulation is implicated in DMD cardiomyopathy. Calcium is primarily removed from the cytosol by the sarcoendoplasmic reticulum calcium ATPase (SERCA). SERCA activity is reduced in DMD. Improving SERCA function may treat DMD cardiomyopathy. Dwarf open reading frame (DWORF) is a recently discovered positive regulator for SERCA, hence, a potential therapeutic target. Methods and Results To study DWORF's involvement in DMD cardiomyopathy, we quantified DWORF expression in the heart of wild-type mice and the mdx model of DMD. To test DWORF gene therapy, we engineered and characterized an adeno-associated virus serotype 9-DWORF vector. To determine if this vector can mitigate DMD cardiomyopathy, we delivered it to 6-week-old mdx mice (6×1012 vector genome particles/mouse) via the tail vein. Exercise capacity, heart histology, and cardiac function were examined at 18 months of age. We found DWORF expression was significantly reduced at the transcript and protein levels in mdx mice. Adeno-associated virus serotype 9-DWORF vector significantly enhanced SERCA activity. Systemic adeno-associated virus serotype 9-DWORF therapy reduced myocardial fibrosis and improved treadmill running, electrocardiography, and heart hemodynamics. Conclusions Our data suggest that DWORF deficiency contributes to SERCA dysfunction in mdx mice and that DWORF gene therapy holds promise to treat DMD cardiomyopathy.

An Integrated Approach for Microprotein Identification and Sequence Analysis.

Next-generation sequencing (NGS) has propelled the field of genomics forward and produced whole genome sequences for numerous animal species and model organisms. However, despite this wealth of sequence information, comprehensive gene annotation efforts have proven challenging, especially for small proteins. Notably, conventional protein annotation methods were designed to intentionally exclude putative proteins encoded by short open reading frames (sORFs) less than 300 nucleotides in length to filter out the exponentially higher number of spurious noncoding sORFs throughout the genome. As a result, hundreds of functional small proteins called microproteins (<100 amino acids in length) have been incorrectly classified as noncoding RNAs or overlooked entirely. Here we provide a detailed protocol to leverage free, publicly available bioinformatic tools to query genomic regions for microprotein-coding potential based on evolutionary conservation. Specifically, we provide step-by-step instructions on how to examine sequence conservation and coding potential using Phylogenetic Codon Substitution Frequencies (PhyloCSF) on the user-friendly University of California Santa Cruz (UCSC) Genome Browser. Additionally, we detail steps to efficiently generate multiple species alignments of identified microprotein sequences to visualize amino acid sequence conservation and recommend resources to analyze microprotein characteristics, including predicted domain structures. These powerful tools can be used to help identify putative microprotein-coding sequences in noncanonical genomic regions or to rule out the presence of a conserved coding sequence with translational potential in a noncoding transcript of interest.

1H NMR global diatropicity in copper hydride complexes.

Understanding the magnetic response of electrons in nanoclusters is essential to interpret their NMR spectra thereby providing guidelines for their synthesis towards various target applications. Here, we consider two copper hydride clusters that have applications in hydrogen storage and release under standard temperature and pressure. Through Born-Oppenheimer molecular dynamics simulations, we study dynamics effects and their contributions to the NMR peaks. Finally, we examine the electrons' magnetic response to an applied external magnetic field using the gauge-including magnetically induced currents theory. Local diatropic currents are generated in both clusters but an interesting global diatropic current also appears. This diatropic current has contributions from three µ3-H hydrides and six Cu atoms that form a chain together with three S atoms from the closest ligands resulting in a higher shielding of these hydrides' 1H NMR response. This explains the unusual upfield chemical shift compared to the common downfield shift in similarly coordinated hydrides both observed in previous experimental reports.

Effect of Biofeedback on the Anxiety of Amateur Athletes / Efecto de la biorretroalimentación sobre la ansiedad de los deportistas aficionados

Biofeedback is an effective strategy to decrease levels of activation and anxiety. The purpose of this study is to determine the degree to which amateur athletes are able to learn to control their autonomic responses through biofeedback. The hypothesis of this study is that amateur athletes with biofeedback intervention will significantly decrease their level of psychophysiological activation compared to those without the intervention. Sixteen amateur marathon athletes participated, from the city of Andria de la Puglia, Italy. They were randomly assigned to two groups, one control and one experimental (i.e., biofeedback condition, which used peripheral temperature, skin conductance, and heart rate techniques). During the intervention period, there were six 15-minute individual sessions over two weeks. Anxiety was evaluated using the STAI survey, which is a subjective scale of activation and a psychophysiological profile. The statistical analysis used in this study is analysis of variance of repeated measurements was used with a significance level of .05. For the Post Hoc or a posteriori comparison, the Sidák and the Bonferroni procedure were conducted. A significant interaction was found between the evaluation conditions, timepoints, group, including peripheral temperature and in the skin conductance. The experimental group has significant lower activation compared to the control group, higher increase of the peripheral temperature and lower conductance. There is no difference in anxiety measured with the STAI. In conclusion, the biofeedback group has learned to control their autonomic responses, as indicated by a significant decrease in the level of psychophysiological activation, compared to the group without intervention.(AU)

Analysis of wavelength-scale 1D depth-dependent refractive-index gradients at an interface by their effects on the internal reflectance near the critical angle.

Light's internal reflectivity near a critical angle is very sensitive to the angle of incidence and the optical properties of the external medium near the interface. Novel applications in biology and medicine of subcritical internal reflection are being pursued. In many practical situations, the refractive index of the external medium may vary with respect to its bulk value due to different physical phenomena at surfaces. Thus, there is a pressing need to understand the effects of a refractive-index gradient at a surface for near-critical-angle reflection. In this work, we investigate theoretically the reflectivity near the critical angle at an interface with glass assuming the external medium has a continuous depth-dependent refractive index. We present graphs of the internal reflectivity as a function of the angle of incidence, which exhibit the effects of a refractive-index gradient at the interface. We analyze the behavior of the reflectivity curves before total internal reflection is achieved. Our results provide insight into how one can recognize the existence of a refractive-index gradient at the interface and shed light on the viability of characterizing it.

Cubic aromaticity in ligand-stabilized doped Au superatoms.

The magnetic response of valence electrons in doped gold-based M@Au8L8 q superatoms (M = Pd, Pt, Ag, Au, Cd, Hg, Ir, and Rh; L = PPh3; and q = 0, +1, +2) is studied by calculating the gauge including magnetically induced currents (GIMIC) in the framework of the auxiliary density functional theory. The studied systems include 24 different combinations of the dopant, total cluster charge, and cluster structure (cubic-like or oblate). The magnetically induced currents (both diatropic and paratropic) are shown to be sensitive to the atomic structure of clusters, the number of superatomic electrons, and the chemical nature of the dopant metal. Among the cubic-like structures, the strongest aromaticity is observed in Pd- and Pt-doped M@Au8L8 0 clusters. Interestingly, Pd- and Pt-doping increases the aromaticity as compared to a similar all-gold eight-electron system Au9L8 +1. With the recent implementation of the GIMIC in the deMon2k code, we investigated the aromaticity in the cubic and butterfly-like M@Au8 core structures, doped with a single M atom from periods 5 and 6 of groups IX-XII. Surprisingly, the doping with Pd and Pt in the cubic structure increases the aromaticity compared to the pure Au case not only near the central atom but encompassing the whole metallic core, following the aromatic trend Pd > Pt > Au. These doped (Pd, Pt)@Au8 nanoclusters show a closed shell 1S21P6 superatom electronic structure corresponding to the cubic aromaticity rule 6n + 2.

Magnetically induced currents and aromaticity in ligand-stabilized Au and AuPt superatoms.

Understanding magnetically induced currents (MICs) in aromatic or metallic nanostructures is crucial for interpreting local magnetic shielding and NMR data. Direct measurements of the induced currents have been successful only in a few planar molecules but their indirect effects are seen in NMR shifts of probe nuclei. Here, we have implemented a numerically efficient method to calculate gauge-including MICs in the formalism of auxiliary density functional theory. We analyze the currents in two experimentally synthesized gold-based, hydrogen-containing ligand-stabilized nanoclusters [HAu9(PPh3)8]2+ and [PtHAu8(PPh3)8]+. Both clusters have a similar octet configuration of Au(6s)-derived delocalized "superatomic" electrons. Surprisingly, Pt-doping in gold increases the diatropic response of the superatomic electrons to an external magnetic field and enhances the aromaticity of [PtHAu8(PPh3)8]+. This is manifested by a stronger shielding of the hydrogen proton in the metal core of the cluster as compared to [HAu9(PPh3)8]2+, causing a significant upfield shift in agreement with experimental proton NMR data measured for these two clusters. Our method allows the determination of local magnetic shielding properties for any component in large 3D nanostructures, opening the door for detailed interpretation of complex NMR spectra.

Centruroides margaritatus scorpion complete venom exerts cardiovascular effects through alpha-1 adrenergic receptors.

Centruroides margaritatus scorpion stings are common in Colombia. However, the cardiovascular toxicity of the venom has not been clarified. AIM: To study the effect and mechanisms of action of the complete venom of C. margaritatus (CmV) on the murine cardiovascular system. METHODS: We evaluated the in vivo effect of CmV LD50 on the mean arterial pressure (MABP), heart rate, and surface electrocardiogram in male adult normotensive Wistar rats. Ex vivo, we evaluated the vascular reactivity of rat aortic rings to increasing concentrations (1 to 60 µg/mL) of CmV using the blockers L-NAME, indomethacin, seratrodast, and prazosin. RESULTS: In the first hour of poisoning, CmV increased the MABP. In the second hour after poisoning, the heart rate decreased as the normalized PR interval and QT corrected increased. After that, cardiovascular shock was demonstrated by a drastic fall in the MABP and signs of cardiac conduction system block. In aortic rings, CmV caused a direct vasoconstrictor effect mediated by alpha-1 adrenergic receptors and counteracted by nitric oxide. CONCLUSION: The direct vascular and probably the cardiac alpha-1 effects likely explain the transient hypertension and the maintenance of cardiac function, while interval lengthening may be due to K+ channel blockage. Afterwards, the effects of both the alpha-1 pathway and the K+ channel pathway converged, resulting in fatal cardiovascular shock. This knowledge could aid in understanding the dynamics of the effects of the venom and in designing treatments to address its cardiovascular effects.

COVID-19: Is There Evidence for the Use of Herbal Medicines as Adjuvant Symptomatic Therapy?

BACKGROUND: Current recommendations for the self-management of SARS-Cov-2 disease (COVID-19) include self-isolation, rest, hydration, and the use of NSAID in case of high fever only. It is expected that many patients will add other symptomatic/adjuvant treatments, such as herbal medicines. AIMS: To provide a benefits/risks assessment of selected herbal medicines traditionally indicated for "respiratory diseases" within the current frame of the COVID-19 pandemic as an adjuvant treatment. METHOD: The plant selection was primarily based on species listed by the WHO and EMA, but some other herbal remedies were considered due to their widespread use in respiratory conditions. Preclinical and clinical data on their efficacy and safety were collected from authoritative sources. The target population were adults with early and mild flu symptoms without underlying conditions. These were evaluated according to a modified PrOACT-URL method with paracetamol, ibuprofen, and codeine as reference drugs. The benefits/risks balance of the treatments was classified as positive, promising, negative, and unknown. RESULTS: A total of 39 herbal medicines were identified as very likely to appeal to the COVID-19 patient. According to our method, the benefits/risks assessment of the herbal medicines was found to be positive in 5 cases (Althaea officinalis, Commiphora molmol, Glycyrrhiza glabra, Hedera helix, and Sambucus nigra), promising in 12 cases (Allium sativum, Andrographis paniculata, Echinacea angustifolia, Echinacea purpurea, Eucalyptus globulus essential oil, Justicia pectoralis, Magnolia officinalis, Mikania glomerata, Pelargonium sidoides, Pimpinella anisum, Salix sp, Zingiber officinale), and unknown for the rest. On the same grounds, only ibuprofen resulted promising, but we could not find compelling evidence to endorse the use of paracetamol and/or codeine. CONCLUSIONS: Our work suggests that several herbal medicines have safety margins superior to those of reference drugs and enough levels of evidence to start a clinical discussion about their potential use as adjuvants in the treatment of early/mild common flu in otherwise healthy adults within the context of COVID-19. While these herbal medicines will not cure or prevent the flu, they may both improve general patient well-being and offer them an opportunity to personalize the therapeutic approaches.

Optical reflectivity of an interface with random refractive-index-contrast patterns.

We develop simple models for the optical reflectivity of an interface in optical contact with random media consisting of discrete volumes of arbitrary form and different refractive indices. Examples of interest are surfaces sprinkled with microdroplets or an interface with biological cells adhered to it at random locations. We focus our attention to the case of internal reflectivity, in which the incidence medium has a larger refractive index than the refractive indices at the other side of the interface. Assuming an incident plane wave, we provide simple approximate expressions for the surface's coherent reflectance and for the surface's total reflectance. We compare predictions of the surface coherent-reflectance model with numerical simulations. Then we use the surface's reflectance models to interpret experimental measurements obtained with an optical prism and a thin vegetable tissue adhered to its base. In general, the surface reflectivity can be used to determine fractional contact area between the interface and microdroplets or biological cells and infer their refractive indices with an accuracy of about 0.5%.

Solubility-Driven Isolation of a Metastable Nonagold Cluster with Body-Centered Cubic Structure.

The conventional synthetic methodology for atomically precise gold nanoclusters by using reduction in solution offers only the thermodynamically most stable nanoclusters. Herein, a solubility-driven isolation strategy is reported to access a metastable gold cluster. The cluster, with the composition of [Au9 (PPh3 )8 ]+ (1), displays an unusual, nearly perfect body-centered cubic (bcc) structure. As revealed by ESI-MS and UV/Vis measurements, the cluster is metastable in solution and converts to the well-known [Au11 (PPh3 )8 Cl2 ]+ (2) within just 90 min. DFT calculations revealed that although both 1 and 2 are eight-electron superatoms, there is a driving force to convert 1 to 2 as shown by the increased cohesion and larger HOMO-LUMO energy gap of 2. The isolation and crystallization of the metastable gold cluster were achieved in a biphasic reaction system in which reduction of gold precursors and crystallization of 1 took place concurrently. This synthetic protocol represents a successful strategy for investigations of other metastable species in metal nanocluster chemistry.

A Compound Isolated from Phyllanthus tenellus Demonstrates Metabolic and Vascular Effects In Vitro.

Common chronic conditions such as metabolic syndrome and diabetes are increasingly associated to metabolic and cardiovascular complications. Although Phyllanthus tenellus leaves have been used in decoctions as a popular remedy to control blood glucose levels and hypertension, its use needs a scientific basis. This study was therefore undertaken to report a phytochemical analysis of P. tenellus leaves and to test if the main active compound has potential to simultaneously tackle several pathophysiological features of metabolic syndrome and diabetes-related metabolic and vascular disorders such as hyperglycaemia, increased platelet activation, and endothelial dysfunction. We performed a partition of the methanolic extract of P. tenellus leaves among different organic solvents followed by chromatographic separation guided by the rat liver microsomal glucose-6-phosphatase assay. Two known tannins were identified by spectroscopic methods as pinocembrin-7-O-[3″-O-galloyl-4″,6″-(S)-hexahydroxydiphenoyl]-α-D-glucose, named P7OG by us, and gemin D. The structural determination of the isolated compounds was based on spectral data. The ability of the main active component, P7OG, to inhibit human platelet aggregation and to modify vascular reactivity of rat aortic rings incubated with high glucose (D-glucose 55 mM) was then evaluated. P7OG was further able to inhibit platelet aggregation induced by adenosine 5'-diphosphate and collagen, showed vasorelaxant effects in arteries precontracted with phenylephrine, and reverted the endothelium-dependent impairment effect of high glucose in rat aortic rings. In conclusion, one tannin isolated from P. tenellus showed promising metabolic, antiaggregant, and vascular effects, which suggests the potential beneficial use of P. tenellus to tackle complex cardiometabolic diseases.

Pomolic acid reduces contractility and modulates excitation-contraction coupling in rat cardiomyocytes.

Pomolic acid (PA) isolated from Licania pittieri has hypotensive effects in rats, inhibits human platelet aggregation and elicits endothelium-dependent relaxation in rat aortic rings. The present study was designed to investigate the effects of PA on cardiomyocytes. Trabeculae and enzymatically isolated cardiomyocytes from rats were used to evaluate the concentration-dependent effects of PA on cardiac muscle tension and excitation-contraction coupling (ECC) by recording Ca2+ transients reported with Fluo-3 and Fura-2, as well as L-type Ca2+ currents (LTCC). PA reduced the contractile force in rat cardiac trabeculae with an EC50 = 14.3 ±â€¯2.4 µM. PA also reduced the amplitude of Ca2+ transients in a concentration-dependent manner, with an EC50 = 10.5 ±â€¯1.3 µM, without reducing sarcoplasmic reticulum (SR) Ca2+ loading. PA decreased the half width of the Ca2+ transient by 31.7 ±â€¯3.3% and increased the decay time and decay time constant (τ) by 7.6 ±â€¯2.7% and 75.6 ±â€¯3.7%, respectively, which was associated with increased phospholamban (PLN) phosphorylation. PA also reversibly reduced the macroscopic LTCC in the cardiomyocyte membrane, but did not demonstrate any effects on skeletal muscle ECC. In conclusion, PA reduces LTCC, Ca2+ transients and cardiomyocyte force, which along with its vasorelaxant effects explain its hypotensive properties. Increased PLN phosphorylation protected the SR from Ca2+ depletion. Considering the effects of PA on platelet aggregation and the cardiovascular system, we propose it as a new potential, multitarget cardiovascular agent with a demonstrated safety profile.

Analytical modeling of optical reflectivity of random plasmonic nano-monolayers.

In this paper, we compare three different models that have been used to interpret reflectivity measurements of supported monolayers of nanoparticles. Two of them: (i) isotropic Maxwell Garnett and (ii) anisotropic two-dimensional-dipolar model are based on an effective-medium approach, while the third one (iii) coherent-scattering model, lies within the framework of multiple-scattering theory. First, we briefly review, on physical grounds, the foundations of each model and write down the corresponding formulas for the calculation of the reflectivity. In the two-dimensional-dipolar model, the dilute limit of the pair-correlation function (also called hole-correlation function) is always used in the calculation of the effective optical response. Then we use these formulas to plot and analyze graphs of the reflectivity of a monolayer of gold nanoparticles on a glass substrate, as a function of several relevant parameters, for two different commonly used experimental configurations. Finally, we discuss the importance of our results and how they can be used to infer the limits of validity of each model.

The norpurpureine alkaloid from Annona purpurea inhibits human platelet activation in vitro.

BACKGROUND: The leaves of Annona purpurea have yielded several alkaloids with anti-aggregation activities against rabbit platelets. This is promising in the search for agents that might act against platelets and reduce the incidence of cardiovascular diseases. Since significant differences in platelet function have been reported between human and animal platelets, a study focusing on the effect of A. purpurea extracts against human platelet activation is necessary. METHODS: The compounds in an A. purpurea ethanolic extract underwent bio-guided fractionation and were used for in vitro human platelet aggregation assays to isolate the compounds with anti-platelet activity. The bioactive compounds were identified by spectroscopic analysis. Additional platelet studies were performed to characterize their action as inhibitors of human platelet activation. RESULTS: The benzylisoquinoline alkaloid norpurpureine was identified as the major anti-platelet compound. The IC50 for norpurpureine was 80 µM against platelets when stimulated with adenosine 5'-diphosphate (ADP), collagen and thrombin. It was pharmacologically effective from 20 to 220 µM. Norpurpureine (220 µM) exhibited its in vitro effectiveness in samples from 30 healthy human donors who did not take any drugs during the 2 weeks prior to the collection. Norpurpureine also gradually inhibited granule secretion and adhesion of activated platelets to immobilized fibrinogen. At the intra-platelet level, norpurpureine prevented agonist-stimulated calcium mobilization and cAMP reduction. Structure-activity relationship analysis indicates that the lack of a methyl group at the nitrogen seems to be key in the ability of the compound to interact with its molecular target. CONCLUSION: Norpurpureine displays a promising in vitro pharmacological profile as an inhibitor of human platelet activation. Its molecular target could be a common effector between Ca2+ and cAMP signaling, such as the PLC-PKC-Ca2+ pathway and PDEs. This needs further evaluation at the protein isoform level.