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Philippe Bernard1, Thomas Scior2, Quoc Tuan Do11Greenpharma SAS, Orléans, France; 2Pharmacy Department, Benemerita Universidad Autonoma de Puebla, Puebla, MexicoAbstract: In men, the level of testosterone decreases with age. At the skin level, the result is observed as a decrease in density and in a lower elasticity. Identifying compounds that are able to increase the level of testosterone appears to be an attractive strategy to develop new antiaging bioactive ingredients for men. Reverse pharmacognosy was successfully applied to identify new natural compounds able to modulate testosterone levels. Among several in silico hits, honokiol was retained as a candidate as it has the greatest potential to become an active ingredient. This result was then validated in vitro on aromatase and 5-alpha-reductase type 1 and 2, which are two types of enzymes implicated in the degradation of free testosterone. Indeed, honokiol was identified as an inhibitor of aromatase, with a half-maximal inhibitory concentration (IC50) of about 50 µM. In addition, honokiol was shown to be an inhibitor of 5-alpha-reductase type 1, with an IC50 of about 75 µM. Taken together, these data indicate that honokiol modulates testosterone levels, and its structure has the potential to serve as a lead for future designs of highly selective inhibitors of 5-alpha-reductase type 1.Keywords: reverse pharmacognosy, honokiol, testosterone, man cosmetics, dermopharmacy

Silica encapsulation under ambient conditions is commonly used to shield protein-based nanosystems from chemical stress. However, encapsulation-induced photo- and structural instabilities at elevated temperatures have been overlooked. Using bovine serum albumin-capped fluorescent gold nanoclusters (BSA-AuNCs) as a model, we demonstrated that chaperone/polymer layer-by-layer complexation can stabilize the template to resist encapsulation-induced fragmentation/reorganization and emission increases at 37 °C or higher temperatures. We first wrapped BSA-AuNCs with α-crystallin chaperones (α-Crys) to gain the highest thermal stability at a 1:50 molar ratio and then enfolded BSA-AuNC/α-Crys with thermoresponsive poly- N -isopropylacrylamide (PNIPAM) at 60 °C to shield silica interaction and increase the chaperone-client protein accessibility. The resulting BSA-AuNC/α-Crys/PNIPAM (BαP) was encapsulated by a sol-gel process to yield BαP-Si (∼80 ± 4.5 nm), which exhibited excellent structural integrity and photostability against chemical and thermal stresses. Moreover, targeted BαP-Si demonstrated prolonged fluorescence stability for cancer cell imaging. This template stabilization strategy for silica encapsulation is biocompatible and applicable to other protein-based nanosystems.

Background: Atopic dermatitis has a marked economic impact and affects the quality of life. A cosmetic compound with an innovative strategy is proposed here as a small chemical neutraligand, GPN279 (previously identified as a theophylline derivative), that binds and potently neutralizes the TARC/CCL17 chemokine, activating the Th2 cell-expressed CCR4 receptor., Objective: Our objective was to evaluate the safety and activity of topically applied GPN279 in mild-to-moderate atopic dermatitis patients in a randomized, double-blind, placebo-controlled, parallel group trial. Such cosmetic active ingredient targeting dry skin with an atopic tendency would open a parallel strategy to the pharmaceutical approach, in particular for mild to moderate subjects, as an alternative to reduce the evolution towards severe forms of atopy., Methods: This 4-week trial included adults with mild-to-moderate atopic dermatitis, according to the SCORAD index. Patients were randomized into two groups treated by topical applications of either an emulsion containing 0.44% GPN279 in placebo on skin lesions or the placebo (4.56% glycerin). Clinical activity was evaluated with the SCORAD as the primary objective. As secondary objectives, POEM, erythema, skin moisturization, its barrier function (TEWL) and safety were evaluated., Results: Twenty-one patients in each group completed the study. SCORAD was significantly improved in the GPN279 group vs. placebo. GPN279 also significantly improved POEM, induced a rapid and significant decrease of erythema, and improved skin moisture. GPN279 and placebo were well tolerated throughout the study., Conclusion: A cosmetic cream comprising the CCL17 neutraligand GPN279 improved the skin barrier and physiology criteria in patients with mild-to-moderate atopic dermatitis., (© 2024 Society of Cosmetic Scientists and the Société Française de Cosmétologie.)

This study aims to use computers to detect and recognize ventilation objects (masks and tubes) and their positions on the patient's face. We created two models: the You Only Look Once (YOLO) and the Transfer Learning (TL) models, to perform this computer vision task. The development processes and comparison of performance will be described in this paper. The TL model had a better performance (93%) compared to the YOLO model (93%). Clinical Relevance- Healthcare providers and researchers interested in the field of computer vision applied in medicine, specifically automatic object detection using video streams or real-time video streaming may benefit from findings reported.

In lower-income countries, the economic contractions that accompany lockdowns to contain COVID-19 transmission can increase child mortality, counteracting the mortality reductions achieved by the lockdown. To formalize and quantify this effect, we build a macrosusceptible-infected-recovered model that features heterogeneous agents and a country-group-specific relationship between economic downturns and child mortality and calibrate it to data for 85 countries across all income levels. We find that in some low-income countries, a lockdown can produce net increases in mortality. The optimal lockdown that maximizes the present value of aggregate social welfare is shorter and milder in poorer countries than in rich ones., (© (2022) by the Economics Department of the University of Pennsylvania and the Osaka University Institute of Social and Economic Research Association.)

Aims: The cosmetic industry needs new preservatives that are effective, natural, safe, cost effective, sustainable and compliant with regulatory standards. This necessity has posed challenges requiring obligations, bioinformatics and bioinspiration as driving forces., Methods and Results: Twenty natural extracts were selected from the Greenpharma Database with parameter filters corresponding to development constraints and antimicrobial properties. We confirmed using minimum inhibition concentration (MIC) assays that eight of the extracts have good bactericidal properties and that one has a high antifungal activity. The latter was purified hinokitiol, a bioproduct from Aomori Hiba wood. This substance provides high resistance against putrefaction; for instance, old Japanese temples were made of Aomori Hiba wood. The combination of hinokitiol with levulinic acid, another bioproduct, demonstrated complementary antimicrobial activities and synergistic effects in MIC studies and measurements according to Kull synergy index. Further, the mixture Hinoline® was tested at 2% in challenge tests and fulfilled criteria A of different standards. It also exerted complementary preservative effects with potassium sorbate and beneficial effects in unbalanced skin microbiota., Conclusion: Hinoline, a new effective preservative from renewable bioresources, was developed., Significance and Impact of the Study: This study accelerates the development of a preservative solution for cosmetics selected from Greenpharma Database, through bioinspiration and the identification of cost-effective investments and resources., (© 2021 The Society for Applied Microbiology.)

Management of respiratory conditions relies on timely diagnosis and institution of appropriate management. Computerized analysis and classification of breath sounds has a potential to enhance reliability and accuracy of diagnostic modality while making it suitable for remote monitoring, personalized uses, and self-management uses. In this paper, we describe and compare sound recognition models aimed at automatic diagnostic differentiation of healthy persons vs patients with COPD vs patients with pneumonia using deep learning approaches such as Multi-layer Perceptron Classifier (MLPClassifier) and Convolutional Neural Networks (CNN).Clinical Relevance-Healthcare providers and researchers interested in the field of medical sound analysis, specifically automatic detection/classification of auscultation sound and early diagnosis of respiratory conditions may benefit from this paper.

Thymus and Activation-Regulated Chemokine (TARC/CCL17) and Macrophage-Derived Chemokine (MDC/CCL22) are two key chemokines exerting their biological effect via binding and activating a common receptor CCR4, expressed at the surface of type 2 helper T (Th2) cells. By recruiting Th2 cells in the dermis, CCL17 and CCL22 promote the development of inflammation in atopic skin. The aim of this research was to develop a plant extract whose biological properties, when applied topically, could be beneficial for people with atopic-prone skin. The strategy which was followed consisted in identifying ligands able to neutralize the biological activity of CCL17 and CCL22. Thus, an in silico molecular modeling and a generic screening assay were developed to screen natural molecules binding and blocking these two chemokines. N -Feruloylserotonin was identified as a neutraligand of CCL22 in these experiments. A cornflower extract containing N -feruloylserotonin was selected for further in vitro tests: the gene expression modulation of inflammation biomarkers induced by CCL17 or CCL22 in the presence or absence of this extract was assessed in the HaCaT keratinocyte cell line. Additionally, the same cornflower extract in another vehicle was evaluated in parallel with N -feruloylserotonin for cyclooxygenase-2 (COX-2) and 5-lipoxygenase (5-LOX) enzymatic cellular inhibition. The cornflower extract was shown to neutralize the two chemokines in vitro , inhibited COX-2 and 5-LOX, and demonstrated anti-inflammatory activities due mainly to the presence of N -feruloylserotonin. Although these findings would need to be confirmed in an in vivo study, the in vitro studies lay the foundation to explain the benefits of the cornflower extract when applied topically to individuals with atopic-prone skin., Competing Interests: The authors declare that there is no conflict of interest regarding the publication of this paper., (Copyright © 2021 Christophe Carola et al.)

Neutrophil gelatinase-associated lipocalin (NGAL) is involved in cardiovascular and renal diseases. Gene inactivation of NGAL blunts the pathophysiological consequences of cardiovascular and renal damage. We aimed to design chemical NGAL inhibitors and investigate its effects in experimental models of myocardial infarction (MI) and chronic kidney disease induced by 5/6 nephrectomy (CKD) on respectively 8-12 weeks old C57Bl6/j and FVB/N male mice. Among the 32 NGAL inhibitors tested, GPZ614741 and GPZ058225 fully blocked NGAL-induced inflammatory and profibrotic markers in human cardiac fibroblasts and primary mouse kidney fibroblasts. The administration of GPZ614741 (100 mg/kg/day) for three months, was able to improve cardiac function in MI mice and reduced myocardial fibrosis and inflammation. The administration of GPZ614741 (100 mg/kg/day) for two months resulting to no renal function improvement but prevented the increase in blood pressure, renal tubulointerstitial fibrosis and profibrotic marker expression in CKD mice. In conclusion, we have identified new compounds with potent inhibitory activity on NGAL-profibrotic and pro-inflammatory effects. GPZ614741 prevented interstitial fibrosis and dysfunction associated with MI, as well as tubulointerstitial fibrosis in a CKD model. These inhibitors could be used for other diseases that involve NGAL, such as cancer or metabolic diseases, creating new therapeutic options.

Catechol estrogens (CEs) are known to be toxic metabolites and the initiators of the oncogenesis of breast cancers via forming covalent adducts with DNAs. CEs shall also react with proteins, but their cellular protein targets remain unexplored. Here, we reported the identification of protein targets of CEs in the soluble cytosol of estrogen-sensitive breast cancer cells by multiple comparative proteomics using liquid chromatography-tandem mass spectrometry (LC-MS/MS) coupled with an improved click chemistry-based workflow. Multiple comparative proteomics composed of an experimental pair (probe versus solvent) and two control pairs (solvent versus solvent and probe versus solvent without enrichment) were studied using stable isotope dimethyl labeling. The use of 4-hydroxyethynylestradiol (4OHEE2) probe with an amide-free linker coupled with on-bead digestion and redigestion of the proteins cleaved from the beads was shown to greatly improve the recovery and identification of CE-adducted peptides. A total of 310 protein targets and 40 adduction sites were repeatedly ( n ≥ 2) identified with D/H (probe/solvent) ratio >4 versus only one identified with D/H >4 from the two control pairs, suggesting that our workflow imposes only a very low background. Meanwhile, multiple comparative D/H ratios revealed that CEs may downregulate many target proteins involved in the metabolism or detoxification, suggesting a negative correlation between CE-induced adduction and expression of proteins acting on the alleviation of stress-induced cellular damages. The reported method and data will provide opportunities to study the progression of estrogen metabolism-derived diseases and biomarkers.

Objective: Video recording and video recognition (VR) with computer vision have become widely used in many aspects of modern life. Hospitals have employed VR technology for security purposes, however, despite the growing number of studies showing the feasibility of VR software for physiologic monitoring or detection of patient movement, its use in the intensive care unit (ICU) in real-time is sparse and the perception of this novel technology is unknown. The objective of this study is to understand the attitudes of providers, patients, and patient's families toward using VR in the ICU., Design: A 10-question survey instrument was used and distributed into two groups of participants: clinicians (MDs, advance practice providers, registered nurses), patients and families (adult patients and patients' relatives). Questions were specifically worded and section for free text-comments created to elicit respondents' thoughts and attitudes on potential issues and barriers toward implementation of VR in the ICU., Setting: The survey was conducted at Mayo Clinic in Minnesota and Florida., Results: A total of 233 clinicians' and 50 patients' surveys were collected. Both cohorts favored VR under specific circumstances (e.g., invasive intervention and diagnostic manipulation). Acceptable reasons for VR usage according to clinicians were anticipated positive impact on patient safety (70%), and diagnostic suggestions and decision support (51%). A minority of providers was concerned that artificial intelligence (AI) would replace their job (14%) or erode professional skills (28%). The potential use of VR in lawsuits (81% clinicians) and privacy breaches (59% patients) were major areas of concern. Further identified barriers were lack of trust for AI, deterioration of the patient-clinician rapport. Patients agreed with VR unless it does not reduce nursing care or record sensitive scenarios., Conclusion: The survey provides valuable information on the acceptance of VR cameras in the critical care setting including an overview of real concerns and attitudes toward the use of VR technology in the ICU., Competing Interests: None declared., (Thieme. All rights reserved.)

Background: Myocardial fibrosis is a hallmark of cardiac remodeling and functionally involved in heart failure development, a leading cause of deaths worldwide. Clinically, no therapeutic strategy is available that specifically attenuates maladaptive responses of cardiac fibroblasts, the effector cells of fibrosis in the heart. Therefore, our aim was to develop novel antifibrotic therapeutics based on naturally derived substance library screens for the treatment of cardiac fibrosis., Methods: Antifibrotic drug candidates were identified by functional screening of 480 chemically diverse natural compounds in primary human cardiac fibroblasts, subsequent validation, and mechanistic in vitro and in vivo studies. Hits were analyzed for dose-dependent inhibition of proliferation of human cardiac fibroblasts, modulation of apoptosis, and extracellular matrix expression. In vitro findings were confirmed in vivo with an angiotensin II-mediated murine model of cardiac fibrosis in both preventive and therapeutic settings, as well as in the Dahl salt-sensitive rat model. To investigate the mechanism underlying the antifibrotic potential of the lead compounds, treatment-dependent changes in the noncoding RNAome in primary human cardiac fibroblasts were analyzed by RNA deep sequencing., Results: High-throughput natural compound library screening identified 15 substances with antiproliferative effects in human cardiac fibroblasts. Using multiple in vitro fibrosis assays and stringent selection algorithms, we identified the steroid bufalin (from Chinese toad venom) and the alkaloid lycorine (from Amaryllidaceae species) to be effective antifibrotic molecules both in vitro and in vivo, leading to improvement in diastolic function in 2 hypertension-dependent rodent models of cardiac fibrosis. Administration at effective doses did not change plasma damage markers or the morphology of kidney and liver, providing the first toxicological safety data. Using next-generation sequencing, we identified the conserved microRNA 671-5p and downstream the antifibrotic selenoprotein P1 as common effectors of the antifibrotic compounds., Conclusions: We identified the molecules bufalin and lycorine as drug candidates for therapeutic applications in cardiac fibrosis and diastolic dysfunction.

This paper focuses on motion analysis of a coupled unmanned surface vehicle (USV)-umbilical cable (UC)-unmanned underwater vehicle (UUV) system to investigate the interaction behavior between the vehicles and the UC in the ocean environment. For this, a new dynamic modeling method for investigating a multi-body dynamics system of this coupling system is employed. Firstly, the structure and hardware composition of the proposed system are presented. The USV and UUV are modeled as rigid-body vehicles, and the flexible UC is discretized using the catenary equation. In order to solve the nonlinear coupled dynamics of the vehicles and flexible UC, the fourth-order Runge-Kutta numerical method is implemented. In modeling the flexible UC dynamics, the shooting method is applied to solve a two-point boundary value problem of the catenary equation. The interaction between the UC and the USV-UUV system is investigated through numerical simulations in the time domain. Through the computer simulation, the behavior of the coupled USV-UC-UUV system is analyzed for three situations which can occur. In particular, variation of the UC forces and moments at the tow points and the configuration of the UC in the water are investigated.

Drug discovery is a challenging and expensive field. Hence, novel in silico tools have been developed in early discovery stage to identify and prioritize novel molecules with suitable physicochemical properties. In many in silico drug design projects, molecular databases are screened by virtual screening tools to search for potential bioactive molecules. The preparation of the molecules is therefore a key step in the success of well-established techniques such as docking, similarity or pharmacophore searching. We review here the lists of several toolkits used in different steps during the cleaning of molecular databases, integrated within a KNIME workflow. During the first step of the automatic workflow, salts are removed, and mixtures are split to get one compound per entry. Then compounds with unwanted features are filtered. Duplicated entries are then deleted while considering stereochemistry. As a compromise between exhaustiveness and computational time, most distributed tautomers at physiological pH are computed. Additionally, various flags are applied to molecules by using either classical molecular descriptors, similarity search to known libraries or substructure search rules. Moreover, stereoisomers are enumerated depending on the unassigned chiral centers. Then, three-dimensional coordinates, and optionally conformers, are generated. This workflow has been already applied to several drug design projects and can be used for molecular database preparation upon request., (Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.net.)

Abundant blood proteins adducted by active electrophiles are excellent markers to predict the risk of electrophile-induced toxicity. However, detecting endogenously adducted proteins by bottom-up selective (or parallel) reaction monitoring (SRM/PRM) is challenging because of the high variability in sample preparation and detection as well as low adduction levels. Here, we reported a new approach in developing PRM methods by combining intact protein measurement with standard additions to target optimal conditions for detecting catechol estrogens (CEs)-adducted human serum albumin (HSA). Blood serum was added with multiple amounts of CEs to obtain serum standards. Intact protein measurement revealed two linear ranges of adduction levels (adducted-CE/HSA): 0.34-0.42 ( R 2 > 0.94) and 0.81-8.54 ( R 2 > 0.96) against the amount of added CEs, respectively. Six adduction sites were identified by trypsin (K20, C34, K73, K281, H338, K378) or chymotrypsin (K20, C34, K378) digestion. PRM methods targeting all adducted/nonadducted peptide pairs based on chymotrypsin or trypsin digestion were developed, and the data were compared with those obtained by intact protein measurement. Correlation plots indicated that chymotrypsin-PRM leads to poor sensitivity and largely underestimated protein adduction levels. Trypsin-PRM leads to sensitive and highly correlated ( R 2 > 0.91) protein adduction levels with a detection limit below the endogenous level and relative standard deviation <25%. As a proof of concept, clinical serum samples were examined by trypsin-PRM, and a slightly higher adduction level was observed for the obesity group when compared with the healthy group. This is the first report on determining adduction levels of blood proteins for long-term exposure to CEs. The standard addition approach can be generally applied to protein adductomics with resolvable mass increments by intact protein measurement to accelerate the development of bottom-up methods close to the inherent limit.

Marine microorganisms are an invaluable source of novel active secondary metabolites possessing various biological activities. In this study, the extraction and isolation of the marine sediment Penicillium species collected in Vietnam yielded ten secondary metabolites, including sporogen AO-1 ( 1 ), 3-indolecarbaldehyde ( 2 ), 2-[(5-methyl-1,4-dioxan-2-yl)methoxy]ethanol ( 3 ), 2-[(2 R -hydroxypropanoyl)amino]benzamide ( 4 ), 4-hydroxybenzandehyde ( 5 ), chrysogine ( 6 ), 3-acetyl-4-hydroxycinnoline ( 7 ), acid 1H-indole-3-acetic ( 8 ), cyclo (Tyr-Trp) ( 9 ), and 2',3'-dihydrosorbicillin ( 10 ). Their structures were identified by the analysis of 1D and 2D NMR data. Among the isolated compounds, 2-[(5-methyl-1,4-dioxan-2-yl)methoxy]ethanol ( 3 ) showed a strong inhibitory effect against Enterococcus faecalis with a minimum inhibitory concentration value of 32 µg/mL. Both 2-[(2 R -hydroxypropanoyl)amino]benzamide ( 4 ) and 4-hydroxybenzandehyde ( 5 ) selectively inhibited E. coli with minimum inhibitory concentration values of 16 and 8 µg/mL, respectively. 2',3'-Dihydrosorbicillin ( 10 ) potentially inhibited α-glucosidase activity at a concentration of 2.0 mM (66.31%).

The purpose of our present study was to develop a forecasting method that would help asthmatic individuals to take evasive action when the probability of an attack was at THEIR PERSONAL THRESHOLD levels. The results are encouraging. Risk factor analysis helps improve the agent's performance (by allowing it to consider personalized risk score of asthma attack triggers while making a decision and being able to ignore the non-triggers), increasing transparency of deep reinforcement learning in medicine applications (by using the results of analyzing risk factors and its association to take actions), and increase accuracy over time since the association risk factor indicators are also changing over time with more accuracy rate. It also brings the possibility of including population-based health in personalized health, which could support a more efficient self-management of chronic diseases.

Objectives: Childhood malnutrition is major health concern in many low- and middle-income countries, including Vietnam. It was a major risk factor for child mortality and adult ill-health. Malnutrition could increase the risk of serious infections; conversely current diseases also had a negative impact on the growth of child. This study, therefore, examines the prevalence of stunting and underweight among 6-59-month-old outpatient children in District 2 Hospital, Ho Chi Minh City, Vietnam., Methods: A cross-sectional study involved a sample of 225 children aged 6-59 months who were randomly selected from the Outpatient Department in District 2 Hospital. Anthropometric measurements and blood test of children were taken to assess the nutritional status and anaemia. A structured questionnaire was also used to collect mothers' and children's characteristics to examine associated risk factors., Results: The prevalence of stunting, underweight, overweight, and anaemia among children aged 6-59 months was 9.8%, 8.4%, 25.8%, and 30.7%, respectively. Underweight significantly correlated only to having breastfeeding in the first hour after birth (RR: 0.02; 95% CI: 0.01-0.17; p<0.001), while stunting was related to age of starting complementary foods from equal to/more than 6 months (RR=0.70, 95%CI=0.50-0.99, p<0.05) and normal birth weight (RR = 0.29, 95%CI = 0.15-0.56, p<0.001)., Conclusions: This study emphasized the importance of measuring the overall nutritional status for children, who have coexisting infectious diseases and anaemia. The high prevalence of malnutrition and anaemia underlined the need for routine screening as well as treatment of children. Additionally, health information strategies should be focused on young children feeding practices to minimize stunting and underweight.

As the world's most traded commodity, oil production is typically well monitored and analyzed. It also has established links to geopolitics, international relations, and security. Despite this attention, the illicit production, refining, and trade of oil and derivative products occur all over the world and provide significant revenues outside of the oversight and regulation of governments. A prominent manifestation of this phenomenon is how terrorist and insurgent organizations-including the Islamic State group, also known as ISIL/ISIS or Daesh-use oil as a revenue source. Understanding the spatial and temporal variation in production can help determine the scale of operations, technical capacity, and revenue streams. This information, in turn, can inform both security and reconstruction strategies. To this end, we use satellite multi-spectral imaging and ground-truth pre-war output data to effectively construct a real-time census of oil production in areas controlled by the ISIL terrorist group. More broadly, remotely measuring the activity of extractive industries in conflict-affected areas without reliable administrative data can support a broad range of public policy and decisions and military operations.

Catechol estrogens (CEs) are metabolic electrophiles that actively undergo covalent interaction with cellular proteins, influencing molecular function. There is no feasible method to identify their binders in a living system. Herein, we developed a click chemistry-based approach using ethinylestradiol (EE2) as the precursor probe coupled with quantitative proteomics to identify protein targets of CEs and classify their binding strengths. Using in situ metabolic conversion and click reaction in liver microsomes, CEs-protein complex was captured by the probe, digested by trypsin, stable isotope labeled via reductive amination, and analyzed by liquid chromatography-mass spectrometry (LC-MS). A total of 334 liver proteins were repeatedly identified ( n ≥ 2); 274 identified proteins were classified as strong binders based on precursor mass mapping. The binding strength was further scaled by D/H ratio (activity probe/solvent): 259 strong binders had D/H > 5.25; 46 weak binders had 5.25 > D/H > 1; 5 nonspecific binders (keratins) had D/H < 1. These results were confirmed using spiked covalent control (strong binder) and noncovalent control (weak binder), as well as in vitro testing of cytochrome c (D/H = 5.9), which showed covalent conjugation with CEs. Many identified strong binders, such as glutathione transferase, catechol-O-methyl transferase, superoxide dismutase, catalase, glutathione peroxidase, and cytochrome c, are involved in cellular redox processes or detoxification activities. CE conjugation was shown to suppress the superoxide oxidase activity of cytochrome c, suggesting that CEs modification may alter the redox action of cellular proteins. Due to structural similarity and inert alkyne group, EE2 probe is very likely to capture protein targets of CEs in general. Thus, this strategy can be adopted to explore the biological impact of CEs modification in living systems.

Control of asthma is critical for disease management and quality of life. Asthma treatment depends on the patient demographic information (e.g., age), and disease severity, which is determined by: (1) how symptoms affect a patient's daily life, (2) measured lung function, and (3) estimated risk of having an asthma attack. In this paper, we will present the Tensorflow Text Classification (TC) method to classify a patient's asthma severity level. We will also propose a Qlearning method to train an agent through trials and errors to improve the prediction accuracy and create a personalized treatment regimen for asthma patients.

Over the past decades, virtual screening has proved itself to be a valuable asset to identify new bioactive compounds. The vast majority of commonly used techniques can be described in three steps: pre-processing the dataset i. e. small (ligands) and eventually larger (receptors) molecules, execute the method and finally analyse the results. Hence, the preparation of ligands is a critical step for success of commonly used virtual screening approaches such as protein-ligand docking, similarity or pharmacophore search. We present here a new workflow, VSPrep, for the pre-processing of small molecules; it is based on freely accessible tools for academics and is integrated within the KNIME platform. It can be used to perform several chemoinformatics tasks such as molecular database cleaning, tautomer and stereoisomer enumeration, focused library design and conformer generation. Additionally, graphical reports of the results are provided to the user as a convenient analysis tool., (© 2017 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.)

Myocardial fibrosis refers to a variety of quantitative and qualitative changes in the interstitial myocardial collagen network that occur in response to cardiac ischaemic insults, systemic diseases, drugs, or any other harmful stimulus affecting the circulatory system or the heart itself. Myocardial fibrosis alters the architecture of the myocardium, facilitating the development of cardiac dysfunction, also inducing arrhythmias, influencing the clinical course and outcome of heart failure patients. Focusing on myocardial fibrosis may potentially improve patient care through the targeted diagnosis and treatment of emerging fibrotic pathways. The European Commission funded the FIBROTARGETS consortium as a multinational academic and industrial consortium with the primary aim of performing a systematic and collaborative search of targets of myocardial fibrosis, and then translating these mechanisms into individualized diagnostic tools and specific therapeutic pharmacological options for heart failure. This review focuses on those methodological and technological aspects considered and developed by the consortium to facilitate the transfer of the new mechanistic knowledge on myocardial fibrosis into potential biomedical applications., (© 2017 The Authors. European Journal of Heart Failure published by John Wiley & Sons Ltd on behalf of European Society of Cardiology.)

The insect repellent N,N-diethyl-m-toluamide (DEET) has been reported to inhibit AChE (acetylcholinesterase) and to possess potential carcinogenic properties with excessive vascularization. In the present paper, we demonstrate that DEET specifically stimulates endothelial cells that promote angiogenesis which increases tumor growth. DEET activates cellular processes that lead to angiogenesis including proliferation, migration and adhesion. This is associated with an enhancement of NO production and VEGF expression in endothelial cells. M3 silencing or the use of a pharmacological M3 inhibitor abrogates all of these effects which reveals that DEET-induced angiogenesis is M3 sensitive. The experiments involving calcium signals in both endothelial and HEK cells overexpressing M3 receptors, as well as binding and docking studies demonstrate that DEET acts as an allosteric modulator of the M3 receptor. In addition, DEET inhibited AChE which increased acetylcholine bioavailability and binding to M3 receptors and also strengthened proangiogenic effects by an allosteric modulation.

Public academic research sites, private institutions as well as small companies have made substantial contributions to the ongoing development of antidiabetic vanadium compounds. But why is this endeavor not echoed by the globally operating pharmaceutical companies, also known as "Big Pharma"? Intriguingly, today's clinical practice is in great need to improve or replace insulin treatment against Diabetes Mellitus (DM). Insulin is the mainstay therapeutically and economically. So, why do those companies develop potential antidiabetic drug candidates without vanadium (vanadium- free)? We gathered information about physicochemical and pharmacological properties of known vanadium-containing antidiabetic compounds from the specialized literature, and converted the data into explanations (arguments, the "pros and cons") about the underpinnings of antidiabetic vanadium. Some discoveries were embedded in chronological order while seminal reviews of the last decade about the Medicinal chemistry of vanadium and its history were also listed for further understanding. In particular, the concepts of so-called "noncomplexed or free" vanadium species (i.e. inorganic oxido-coordinated species) and "biogenic speciation" of antidiabetic vanadium complexes were found critical and subsequently documented in more details to answer the question.

Excessive signaling by chemokines has been associated with chronic inflammation or cancer, thus attracting substantial attention as promising therapeutic targets. Inspired by chemokine-clearing molecules shaped by pathogens to escape the immune system, we designed a generic screening assay to discover chemokine neutralizing molecules (neutraligands) and unambiguously distinguish them from molecules that block the receptor (receptor antagonists). This assay, called TRIC-r, combines time-resolved intracellular calcium recordings with pre-incubation of bioactive compounds either with the chemokine or the receptor-expressing cells. We describe here the identification of high affinity neutraligands of CCL17 and CCL22, two chemokines involved in the Th2-type of lung inflammation. The decoy molecules inhibit in vitro CCL17- or CCL22-induced intracellular calcium responses, CCR4 endocytosis and human T cell migration. In vivo, they inhibit inflammation in a murine model of asthma, in particular the recruitment of eosinophils, dendritic cells and CD4(+)T cells. Altogether, we developed a successful strategy to discover as new class of pharmacological tools to potently control cell chemotaxis in vitro and in vivo.

Nature was and still is a prolific source of inspiration in pharmacy, cosmetics, and agro-food industries for the discovery of bioactive products. Informatics is now present in most human activities. Research in natural products is no exception. In silico tools may help in numerous cases when studying natural substances: in pharmacognosy, to store and structure the large and increasing number of data, and to facilitate or accelerate the analysis of natural products in regards to traditional uses of natural resources; in drug discovery, to rationally design libraries for screening natural compound mimetics and identification of biological activities for natural products. Here we review different aspects of in silico approaches applied to the research and development of bioactive substances and give examples of using nature-inspiring power and ultimately valorize biodiversity., (Georg Thieme Verlag KG Stuttgart · New York.)

We have designed, synthesized and evaluated homotryptophan analogues as possible mechanism-based inhibitors for Escherichia coli tryptophan indole-lyase (tryptophanase, TIL, E.C. 4.1.99.1). As a quinonoid structure is an intermediate in the reaction mechanism of TIL, we anticipated that homologation of the physiological substrate, L-Trp would provide analogues resembling the transition state for β-elimination, and potentially inhibit TIL. Our results demonstrate that L-homotryptophan (1a) is a moderate competitive inhibitor of TIL, with Ki=67 μM, whereas L-bishomotryptophan (1b) displays more potent inhibition, with Ki=4.7 μM. Pre-steady-state kinetics indicated the formation of an external aldimine and quinonoid with 1a, but only the formation of an external aldimine for 1b, suggesting differences in the inhibition mechanism. These results demonstrate that formation of a quinonoid complex is not required for strong inhibition. In addition, the Trp analogues were evaluated as inhibitors of Salmonella typhimurium Trp synthase. Our results indicate that compound 1b is at least 25-fold more selective toward TIL than Trp synthase. We report that compound 1b is comparable to the most potent inhibitor previously reported, while displaying high selectivity for TIL. Thus, 1b is a potential lead for the development of novel antibacterials., (Copyright © 2014 Elsevier Inc. All rights reserved.)

Generalised bone mineral density (BMD) reduction often occurs in established rheumatoid arthritis (RA); however, in early RA, there is a disagreement with regard to BMD in the femoral neck and lumbar spine, and there is no available information for the whole body. Therefore, the aims of this study were to investigate the BMD, frequency of osteoporosis and the risk factors for BMD reduction in Vietnamese women with early RA. BMD in the femoral neck, lumbar spine L1-4 and whole body was measured in 105 women with early RA (disease duration ≤3 years) and 105 age-matched healthy women (26-73 years) using a dual energy X-ray absorptiometry. Femoral neck and whole body BMD in women with RA were lower (p < 0.05) than controls, while lumbar spine BMD was similar between two groups. The frequency of osteoporosis in the femoral neck, lumbar spine and whole body in women with RA aged ≥50 were higher (p < 0.05) than controls: 41.8% versus 29.5%, 42.2% versus 37.7% and 37.1% versus 28%, respectively. There were associations between the frequencies of osteoporosis at all sites with postmenopausal status, glucocorticoid use, rheumatoid factor positivity and disease activity with lumbar spine BMD and disease disability with femoral neck and whole body BMD. In conclusion, women with early RA had significantly lower femoral neck and whole body BMD, but had similar lumbar spine BMD compared with controls. The frequency of osteoporosis at all sites was significantly higher in women with RA than controls, suggesting that assessment of BMD should be considered in women with early RA.

Objectives: To characterize body composition phenotypes using a dual-energy X-ray absorptiometry (DXA) method and to explore factors potentially contributing to alterations in body composition in Vietnamese women with early RA., Methods: A total of 105 women with early RA (disease duration ≤ 3 years) and 105 age-matched healthy women underwent physical examination, total and regional lean mass (LM) and fat mass (FM) with DXA. The 28-joint DAS (DAS-28) and disability using HAQ score, nutrition, physical activity and medications were recorded., Results: Means of weight and BMI were similar in RA patients and controls, but means of total body and trunk FM in RA patients were higher: 19.1 vs 16.9 kg (P = 0.007) and 10.1 vs 8.1 kg (P = 0.01), respectively, and appendicular LM was lower: 12.9 vs 14.1 kg (P = 0.02). The proportion of unhealthy body composition phenotypes (sarcopenia, overfat and sarcopenic obesity) in RA patients was higher (P < 0.001) than in controls. DAS-28 score was positively correlated with total FM and fat distribution ratio, and HAQ score was inversely correlated with appendicular LM. These body composition changes were associated with RF seropositivity, HAQ and DAS-28 scores., Conclusions: Women with early RA had a significantly higher proportion of unhealthy body composition phenotypes, higher total and truncal FM and lower appendicular LM than controls. Disease activity and disability scores were associated with unhealthy body composition. These findings suggest that clinicians should encourage muscle strengthening and fat loss in RA patients to reduce their disability.

Prior to its total synthesis, a new vanadium coordination compound, called TSAG0101, was computationally designed to inhibit the enzyme protein tyrosine phosphatase 1B (PTP1B). The PTP1B acts as a negative regulator of insulin signaling by blocking the active site where phosphate hydrolysis of the insulin receptor takes place. TSAG001, [V(V)O(2)(OH)(picolinamide)], was characterized by infrared (IR) and nuclear magnetic resonance (NMR) spectroscopy; IR: ν/cm(-1) 3,570 (NH), 1,627 (C=O, coordinated), 1,417 (C-N), 970/842 (O=V=O), 727 δ̣ (pyridine ring); (13)C NMR: 5 bands between 122 and 151 ppm and carbonyl C shifted to 180 ppm; and (1)H NMR: 4 broad bands from 7.6 to 8.2 ppm and NH(2) shifted to 8.8 ppm. In aqueous solution, in presence or absence of sodium citrate as a biologically relevant and ubiquitous chelator, TSAG0101 undergoes neither ligand exchange nor reduction of its central vanadium atom during 24 hours. TSAG0101 shows blood glucose lowering effects in rats but it produced no alteration of basal- or glucose-induced insulin secretion on β cells during in vitro tests, all of which excludes a direct mechanism evidencing the extrapancreatic nature of its activity. The lethal dose (LD(50)) of TSAG0101 was determined in Wistar mice yielding a value of 412 mg/kg. This value is one of the highest among vanadium compounds and classifies it as a mild toxicity agent when compared with literature data. Due to its nonsubstituted, small-sized scaffold design, its remarkable complex stability, and low toxicity; TSAG0101 should be considered as an innovative insulin-mimetic principle with promising properties and, therefore, could become a new lead compound for potential nonpeptide PTP1B inhibitors in antidiabetic drug research. In view of the present work, the inhibitory concentration (IC(50)) and extended solution stability will be tested.

A huge amount of data has been generated by decades of pharmacognosy supported by the rapid evolution of chemical, biological and computational techniques. How can we cope with this overwhelming mass of information? Reverse pharmacognosy was introduced with this aim in view. It proceeds from natural molecules to organisms that contain them via biological assays in order to identify an activity. In silico techniques and particularly inverse screening are key technologies to achieve this goal efficiently. Reverse pharmacognosy allows us to identify which molecule(s) from an organism is(are) responsible for the biological activity and the biological pathway(s) involved. An exciting outcome of this approach is that it not only provides evidence of the therapeutic properties of plants used in traditional medicine for instance, but may also position other plants containing the same active compounds for the same usage, thus increasing the curative arsenal e.g. development of new botanicals. This is particularly interesting in countries where western medicines are still not affordable. At the molecular level, in organisms, families of metabolites are synthesized and seldom have a single structure. Hence, when a natural compound has an interesting activity, it may be desirable to check whether there are more active and/or less toxic derivatives in organisms containing the hit - this corresponds to a kind of "natural combinatorial" chemistry. At a time when the pharmaceutical industry is lacking drug candidates in clinical trials, drug repositioning - i.e. exploiting existing knowledge for innovation - has never been so critical. Reverse pharmacognosy can contribute to addressing certain issues in current drug discovery - such as the lack of clinical candidates, toxicity... - by exploiting existing data from pharmacognosy. This review will focus on recent advances in computer science applied to natural substance research that consolidate the new concept of reverse pharmacognosy.

Introduction: Rheumatoid arthritis (RA) is associated with increased morbidity and mortality due to cardiovascular disease, and this occurs early in the disease process. The metabolic syndrome (MetS) may contribute to the excess cardiovascular burden observed in RA; however, little information is available regarding MetS in early RA. We aimed to identify the prevalence of MetS and to determine the potential factors associated with the presence of MetS in Vietnamese women with early RA., Methods: A total of 105 consecutive women with early RA (disease duration ≤3 years) and 105 age-matched healthy women were checked for MetS according to six MetS definitions (Joint Consensus, International Diabetes Federation, National Cholesterol Education Program 2004 and 2001, European Group for Study of Insulin Resistance, and World Health Organization). Multivariate logistic regression models were constructed to determine independent predictors of MetS in women with RA., Results: Prevalence of MetS varied from 16.2% to 40.9% according to the definitions used in women with RA, and was higher (P < 0.001) than in healthy controls (from 10.5% to 22.9%). Among individual components of MetS, differences between women with RA and controls were observed for hypertension (P < 0.001), low high density lipoprotein-cholesterol (HDL-C) levels (P < 0.001), and abdominal obesity (P = 0.019). After adjusting for age and physical activity, higher erythrocyte sedimentation rate (ESR) (odds ratios (OR) = 1.516, 95% confidence interval (CI): 1.073 to 3.195, P = 0.042), disease activity score (DAS28) (OR = 1.736, 95% CI: 1.293 to 2.786, P = 0.019), health assessment questionnaire (HAQ) score (OR = 1.583, 95% CI: 1.195 to 2.367, P = 0.035), and less methotrexate use (OR = 0.736, 95% CI: 0.547 to 0.962, P = 0.024) remained significant independent predictors of the presence of MetS in women with RA., Conclusions: Women with early RA already had higher prevalence of MetS compared with healthy controls. Higher systemic inflammatory marker, disease activity and disability scores, and less methotrexate use were independent predictors associated with the presence of MetS in women with early RA. These findings suggest that physicians should screen for MetS in women with early RA to control its components and therefore reduce their risk of cardiovascular diseases.

Several small weight indole derivatives (D-64131, D-24851, BPR0L075, BLF 61-3, and ATI derivatives) are potent tubulin polymerization inhibitors and show nanomolar antiproliferative activity. Among them, indolobenzazepin-7-ones were recently disclosed as potent antimitotic agents. In an effort to improve this structure, we prepared new derivatives in order to evaluate their antiproliferative activity. 5,6,7,9-Tetrahydro-8H-indolo[2,3-e][3]benzazocin-8-one (1m) was found to be the most potent derivative inhibiting the cell growth of several cancer cell lines in the lower nanomolar range.

Quantitative Structure-Activity Relationships (QSAR) are based on the hypothesis that changes in molecular structure reflect proportional changes in the observed response or biological activity. In order to successfully conduct QSAR studies certain conditions have to be met that are not frequently reported in the literature. This suggests that some authors are not aware of the principle flaws, occasional shortcomings, and circumstantial downsides of QSAR methods. The present paper focuses on prerequisites to set up correct models and on limitations of model applications. Their implications are systematically described and illustrated as pitfalls that have strong implications in QSAR, and possible solutions are suggested. The paper is focused on small scale 2D- and 3D-QSAR studies for lead optimization. The work is enriched with comprehensive comments and non-mathematical explanations for the computer practitioner in Medicinal Chemistry.

A series of 8-substituted pyrazolo[1,5-a]-1,3,5-triazines were considered as a bioisosteric replacement for the 9-substituted adenine derivatives resulting in the discovery of 8-(2-methoxybenzyl)-4-(N-methylamino)-2-n-propylpyrazolo[1,5-a]-1,3,5-triazine (14d) and 2-trifluoromethyl-8-(2-methoxybenzyl)-4-(N-methylamino)pyrazolo[1,5-a]-1,3,5-triazine (14e) as a new structural class of potent phosphodiesterase type 4 inhibitors (IC(50)=13 nM and 11 nM, respectively) with high isoenzyme selectivity. An original tandem of reactions involving a palladium-mediated cross-coupling reaction (PMCCR) of the readily available 8-iodo-2-methyl-4-(N-methyl-N-phenylamino)pyrazolo[1,5-a]-1,3,5-triazine (11a) and arylboronic acids or alkynes followed by the displacement of the N-methyl-N-phenylamino group constitute the key steps in a novel synthetic approach developed herein. The treatment of 11a-c with n-BuLi and selected aldehydes represents an interesting alternative to the PMCCR for the synthesis of benzylic derivatives 14a-i. Preliminary biological testing has shown that compounds 14d and 14e strongly inhibit LPS-induced TNFalpha release from human mononuclear cells from healthy subjects. These two compounds were selected for further biological evaluation.

Drug repositioning strategy is an interesting approach for pharmaceutical companies; especially to increase their productivity. SELNERGY(tm) is a reverse docking based-program able to virtually screen thousands of compounds on more than 2000 3D biological targets. This program was successfully applied to tofisopam and revealed that the isomers of tofisopam are able to fit with phosphodiesterase 4. This old drug was used as a racemic mixture to treat anxiety in the eighties and was recently shown to act as a PDE4 inhibitor. Thanks to this strategy we demonstrated that tofisopam acts via the inhibition of PDE4 in the submicromolar range. Moreover, we firstly showed that the S-enantiomer of tofisopam is ten times more active than R-enantiomer. The identification of the biochemical mechanism of tofisopam isomers now allows to reposition this drug in new therapeutic indications where modulation of cAMP via PDE4 inhibitors are possible.

Reverse pharmacognosy aims at finding biological targets for natural compounds by virtual or real screening and identifying natural resources that contain the active molecules. We report herein a study focused on the identification of biological properties of meranzin, a major component isolated from Limnocitrus littoralis (Miq.) Swingle. Selnergy, an IN SILICO biological profiling software, was used to identify putative binding targets of meranzin. Among the 400 screened proteins, 3 targets were selected: COX1, COX2 and PPARgamma. Binding tests were realised for these 3 protein candidates, as well as two negative controls. The predictions made by Selnergy were consistent with the experimental results, meaning that these 3 targets can be modulated by an extract containing this compound in a suitable concentration. These results demonstrate that reverse pharmacognosy and its inverse docking component is a powerful tool to identify biological properties for natural molecules and hence for plants containing these compounds.

The relationship between poverty and mental health has received considerable attention in the recent literature. However, the associations presented in existing studies typically rely on limited samples of individuals and on proxy indicators for poverty such as education, the lack of tap water, or being unemployed. We revisit the relationship between poverty and mental health using data from nationally representative household surveys in Bosnia and Herzegovina, Indonesia and Mexico, along with special surveys from India and Tonga. As in previous studies, we find that individuals who are older, female, widowed, and in poor health are more likely to report worse mental health outcomes. Individuals living with others with poor mental health are significantly more likely to report worse mental health themselves. The size of the coefficients and their significance are comparable across the five countries. In contrast to previous studies, the relationship between higher education and better mental health is weak or non-existent. Furthermore, there is no consistent association between consumption poverty and mental health - in two countries mental health measures are marginally worse for the poor; in two countries there is no association; and in one country mental health measures are better for the poor compared to the non-poor. Moreover, the sizes of the coefficients for both education and consumption poverty are small compared to other factors considered here. While the lack of an association between consumption poverty and mental health implies that poor mental health is not a "disease of affluence", neither is it a disease of poverty. Changes in life circumstances brought on, for instance, by illness may have a greater impact on mental health than levels of poverty. Effective public health policy for mental health should focus on protecting individuals and households from adverse events and on targeted interventions following such adverse changes.

Neurokinin A stimulates physiological responses in the peripheral and central nervous systems upon interacting primarily with the tachykinin NK2 receptor (NK2R). In this study, the structure of NKA bound to the NK2R is characterised by use of fluorescence resonance energy transfer. Four fluorescent NKA analogues with Texas red introduced at amino acid positions 1, 4, 7 and 10 were prepared. When bound to a NK2R carrying enhanced green fluorescent protein at the N-terminus, all peptides reduce green fluorescent protein fluorescence from 10% to 50% due to energy transfer. The derived donor-acceptor distances are 46, 55, 59 and 69 A for the fluorophore linked to positions 1-10, respectively. The monotonic increase in distance clearly indicates that the peptide adopts an extended structure when bound to its receptor. The present data are used, in combination with rhodopsin structure, fluorescence studies, photoaffinity labelling and site-directed mutagenesis data to design a computer model of the NKA-NK2R complex. We propose that the N-terminus of NKA is exposed and accessible to the extracellular medium. Subsequent amino acids of the NKA peptide become progressively more buried residues up to approximately one-third of the transmembrane-spanning domain.

Vanadate can be bioequivalent to phosphate and replace it in cellular metabolism. The detection of insulin-like activity has spurred interest in the development of oral anti-diabetic drugs containing vanadium. We collected and evaluated a vast toxicity data set and discussed molecular aspects related to insulin-mimetic effects of vanadium complexes.

The aim of reverse pharmacognosy is to find new biological targets for natural compounds by virtual or real screening and identify natural resources that contain the active molecules. To demonstrate the applicability of this concept, we report here a study on epsilon-viniferin, an active ingredient for cosmetic development. Nevertheless, this natural substance is weakly defined in terms of biological properties. SELNERGY, an inverse docking computer software, was used to identify putative binding biological targets for epsilon-viniferin. Among the 400 screened proteins two targets were retained. For cosmetic application, cyclic nucleotide phosphodiesterase 4 (PDE4) was the most interesting candidate. Moreover, other PDE subtypes (1, 2, 3, 5 and 6) were not retained, indicating a selectivity for PDE4. The experimental binding tests on the 6 subtypes of PDE revealed a significant selectivity of epsilon-viniferin for the PDE4 subtype. This selectivity was confirmed by evaluation of epsilon-viniferin on the secretion of TNF-alpha and Interleukin-8. Our data demonstrated that epsilon-viniferin possesses anti-inflammatory properties by inhibiting PDE4 subtype. In conclusion, reverse pharmacognosy and its inverse docking component cannot only be integrated into a program for new lead discovery but is also a useful approach to find new applications for identified compounds.

To exploit available structural information about the cyclooxygenase enzyme for the virtual screening of large chemical libraries, a docking-scoring protocol was tuned and validated. The screening accuracy was assessed using a series of known inhibitors and a set of diverse a priori inactive compounds that was seeded with known active ligands. The major parameters of the DOCK algorithm were investigated. A large improvement of the results was obtained on tweaking some of them. The generated complexes were rescored using six scoring functions. In this way, the striking importance of this step was demonstrated, as well as the good performances of DOCK energy and SCORE for this target. The results were further improved via a consensus approach. As a first application, a subset of a large compound library was screened using this protocol. Among the compounds that were selected for biological testing, a third of them turned out to have a significant enzyme inhibition.

Combinatorial chemistry and high-throughput screening (HTS) have led to the identification of numerous agents that are active and selective in vitro. Identifying drugs that are active in vivo, however, remains a challenge. Traditional medicinal cures based on natural materials have proven useful for many populations worldwide, representing huge and disperse tracts of knowledge that are sometimes neglected in Western research due to differences in the concepts of illness. In this review we introduce a new approach, termed 'reverse pharmacognosy' (from diverse molecules to plants), which can be coupled with pharmacognosy (from biodiverse plants to molecules). Reverse pharmacognosy utilizes new techniques, such as HTS, virtual screening and a knowledge database containing the traditional uses of plants. Integrating pharmacognosy and reverse pharmacognosy in the research process may provide an efficient and rapid tool for natural drug discovery.

In our efforts to find new active tyrosinase inhibitors for skin-whitening or antibrowning preparations, we investigated 67 tropical plants belonging to 38 families, which were evaluated for their anti-tyrosinase activity. The results of our investigation show that extracts of 5 plants, Stryphnodendron barbatimao, Portulaca pilosa, Cariniana brasiliensis, Entada africana and Prosopis africana present interesting in vitro mushroom tyrosinase inhibition (over 90%), similar to a positive control: Morus alba. These 5 plants will be studied in order to isolate and identify phytochemical compounds, involved in this biological activity.

The structure of two selective inhibitors, Ac-Tyr-Ile-Arg-Ile-Pro-NH2 and Ac-(4-Amino-Phe)-(Cyclohexyl-Gly)-Arg-NH2, in the active site of the blood clotting enzyme factor Xa was determined by using transferred nuclear Overhauser effect nuclear magnetic resonance (NMR) spectroscopy. They represent a family of peptidic inhibitors obtained by the screening of a vast combinatorial library. Each structure was first calculated by using standard computational procedures (distance geometry, simulated annealing, energy minimization) and then further refined by systematic search of the conformation of the inhibitor docked in the active site and repeating the simulated annealing and energy minimization. The final structure was optimized by molecular dynamics simulations of the inhibitor-complex in water. The NMR restraints were kept throughout the refinement. The inhibitors assume a compact, very well defined conformation, embedded into the substrate binding site not in the same way as a substrate, blocking thus the catalysis. The model allows to explain the mode of action, affinity, and specificity of the peptides and to map the active site.

The interaction of Li+ with myo-inositol monophosphatase was studied by 7Li-NMR spectroscopy. Li+ binding to the enzyme induces a downfield shift and broadening of the 7Li-NMR signal. Changes of the chemical shift were used to follow the titration of the enzyme with lithium and to determine a dissociation constant, Kd = (1.0 +/- 0.1) mM. Only one major binding site/enzyme subunit was inferred. The complex forms independently of the presence of inorganic phosphate. Metals from the group IIa of the periodic table compete with Li+ binding with the affinity increasing in the order Mg2+ < Ca2+ < Be2+. In contrast to lithium, their binding is enhanced by phosphate.