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Background: One third of the patients with Major Depression Disorder (MDD) fail to respond to conventional antidepressant treatments - treatment resistant depression (TRD). Low-dose IV ketamine (LDIVK) treatment has become a promising solution for these patients. However, infusion regimens for LDIVK acute series have not been standardized yet. Also, evidence is scarce for retrialing LDIVK treatment to manage MDD symptom recurrences. Methods: This is a case study of a 45-year-old female experiencing MDD TRD, who completed a triweekly and a biweekly acute course (4 week) of LDIVK treatment three years apart. Treatment effectiveness was assessed by comparing baseline to weekly depression severity scores using validated psychiatric scales to determine changes in depression symptoms. Tolerability was assessed by systematically registering side effects through the infusions. Results: In both acute series, the patient achieved symptomatic response equivalent to remission. On the triweekly acute course the patient achieved symptom remission by the second week and in the biweekly acute course by the third week. Reported side effects were mild and transient. Limitations: Results of only one patient included in this report and a different number of LDIVK infusions in each acute course, with the triweekly vs biweekly course providing four additional infusions in four weeks. Conclusions: These results suggest that implementing a triweekly regimen may be well-tolerated and result in a faster treatment response in certain patients, compared to a biweekly regimen. Furthermore, this case also supports the possibility of a ketamine retrial in case of major depressive recurrences.

Competing Interests: Declaration of Competing Interest Dr Gustavo Vazquez has received consulting and speaking honoraria from AbbVie, Allergan, CANMAT, Elea/Phoenix, Eurofarma, Gador, Janssen, Lundbeck, NeonMind Biosciences, Tecnofarma, Raffo, Otsuka, Psicofarma, and Sunovion, and research grants from CAN-BIND, CIHR, PCH, KHSC and Queen's University.

Human and dog health are inextricably linked, and although our relationship with dogs brings numerous benefits for our well-being, it is known that they can transmit disease. Bariloche is a Patagonian tourist city with heterogeneous social composition. This cross-sectional study evaluates the population of free-roaming dogs and their intestinal parasites in relation to the socioeconomic level of the city's human population. Census areas were used as survey units, stratified in three levels according to socioeconomic status. The free-roaming dog population was estimated by walking around each census area. Eight fresh feces per census count area were collected and analyzed using coproparasitological flotation tests, and CoproELISA was used to detect Echinococcus sp. A total of 858 free-roaming dogs were registered along 40.9 km, with significant differences between socioeconomic strata: the highest numbers were found in the lowest income strata. Of the feces collected, 39.2% tested positive for parasites, those associated with a lower socioeconomic level having higher percentages of positive feces and a greater number of species. Eight species of helminths were found, some of which were zoonotic, such as Echinococcus sp., Toxocara canis, and Dibothriocephalus latus. The presence of parasites can be explained by the number of free-roaming dogs per census count area. The free-roaming dogs generally have owners, and their parasitic infection is strongly associated with the socioeconomic level of the population. The main problem is irresponsible pet care, which generates healthy conditions for both dogs and humans. Thus, both dogs and humans deserve effective ethical public policies., (Copyright © 2022 Elsevier B.V. All rights reserved.)

The use of randomized clinical trials, in particular placebo-controlled trials, for drug approval, is the subject of long-standing debate in the scientific community and beyond. This study offers consensus recommendations from clinical and academic experts to guide the selection of clinical trial design in psychiatry. Forty-one highly cited clinical psychiatrists and/or researchers participated in a Delphi survey. Consensus statements were developed based on the findings of a published, peer-reviewed systematic review. Participants evaluated statements in two survey rounds, following the Delphi method. The expert panel achieved consensus on 7 of 21 recommendations regarding the use of randomized clinical trials. The endorsed recommendations were: (i) Results from placebo-controlled trials are the most reliable and (ii) are necessary despite the growing placebo-effect; (iii) it is ethical to enroll patients in placebo-arms when established treatment is available, if there is no evidence of increased health risk; (iv) There is a need to approve new drugs with the same efficacy as existing treatments, but with different side-effect profiles; (v) Non-inferiority trials incur an increased risk of approving ineffective medications; (vi) The risk of approving an ineffective drug justifies trial designs that incur higher costs, and (vii) superiority trials incur the risk of rejecting potentially efficacious treatments. The endorsed recommendations inform the choice of trial-design appropriate for approval of psychopharmacological drugs. The recommendations strongly support the use of randomized clinical trials in general, and the use of placebo-controlled trials in particular., Competing Interests: Declaration of Competing Interest Guy M. Goodwin is a NIHR Emeritus Senior Investigator, holds shares in P1vital and P1Vital products and has served as consultant, advisor or CME speaker in the last 3 years for Beckley Psytech, Clerkenwell Health, Compass pathways, Evapharma, Janssen, Lundbeck, Medscape, Novartis, P1Vital, Sage, Servier. The views expressed are those of the author(s) and not necessarily those of the NHS, the NIHR or the Department of Health. Michael Berk is supported by a NHMRC Senior Principal Research Fellowship (1156072). MB has received Grant/Research Support from the NIH, Cooperative Research Centre, Simons Autism Foundation, Cancer Council of Victoria, Stanley Medical Research Foundation, Medical Benefits Fund, National Health and Medical Research Council, Medical Research Futures Fund, Beyond Blue, Rotary Health, A2 milk company, Meat and Livestock Board, Woolworths, Avant and the Harry Windsor Foundation, has been a speaker for Abbot, Astra Zeneca, Janssen and Janssen, Lundbeck and Merck and served as a consultant to Allergan, Astra Zeneca, Bioadvantex, Bionomics, Collaborative Medicinal Development, Janssen and Janssen, Lundbeck Merck, Pfizer and Servier – all unrelated to this work. Andreas Meyer-Lindenberg has received consultant fees from: Boehringer Ingelheim, Elsevier, Brainsway, Lundbeck Int. Neuroscience Foundation, Lundbeck A/S, The Wolfson Foundation, Bloomfield Holding Ltd, Shanghai Research Center for Brain Science, Thieme Verlag, Sage Therapeutics, v Behring Röntgen Stiftung, Fondation FondaMental, Janssen-Cilag GmbH, MedinCell, Brain Mind Institute, Agence Nationale de la Recherche, CISSN (Catania Internat. Summer School of Neuroscience), Daimler und Benz Stiftung, American Association for the Advancement of Science, Servier International. Additionally, he has received speaker fees from: Italian Society of Biological Psychiatry, Merz-Stiftung, Forum Werkstatt Karlsruhe, Lundbeck SAS France, BAG Psychiatrie Oberbayern, Klinik für Psychiatrie und Psychotherapie Ingolstadt, med Update GmbH, Society of Biological Psychiatry, Siemens Healthineers, Biotest AG -All unrelated to this work. Terence A. Ketter has been a consultant to Otsuka Pharmaceuticals, Sunovion Pharmaceuticals, Abbvie, and Alkermes. Joseph Goldberg has been a consultant to BioXCel, Otsuka, Sage Pharmaceuticals, Sunovion, and WebMD, and served on the speaker boards for Allergan, Intracellular Therapies, and Sunovion. Rif S. El-Mallakh is on the speaker bureau of Alkermes, Eisai, Indivior, Intra-Cellular Therapeutics, Janssen, Lundbeck, Noven, Otsuka, Sunonvion, and Teva. Lakshmi N Yatham has been on speaker/advisory boards for, or has received research grants from Abbvie, Alkermes, Allergan, CANMAT, CIHR, DSP, Merck, and Sanofi Rasmus W. Licht has within the preceding three years served an advisory board of Janssen Cilag and Sagw, and received speaker honorarium from Astra-Zeneca, Jannsen-Cilag, Servier and Lundbeck Allan H. Young has been employed by King's College London; Honorary Consultant SLaM (NHS UK). Young has participated in paid lectures and advisory boards for the following companies with drugs used in affective and related disorders: Astrazenaca, Eli Lilly, Lundbeck, Sunovion, Servier, Livanova, Janssen, Allegan, Bionomics, Sumitomo Dainippon Pharma, COMPASS Allan H. Young's independent research funded by the National Institute for Health Research (NIHR) Biomedical Research Centre at South London and Maudsley NHS Foundation Trust and King's College London. The views expressed are those of the authors and not necessarily those of the NHS, the NIHR, or the Department of Health.Consultant to Johnson & Johnson and Livanova. Received honoraria for attending advisory boards and presenting talks at meetings organized by LivaNova. Prof. Alessandro Serretti is or has been consultant/speaker for: Abbott, Abbvie, Angelini, Astra Zeneca, Clinical Data, Bo- heringer, Bristol Myers Squibb, Eli Lilly, GlaxoSmithKline, Innovapharma, Italfarmaco, Janssen, Lundbeck, Naurex, Pfizer, Polifarma, Sanofi, Servier. Ayşegül Yildiz has nothing to declare. Jean Michel Azorin has received honoraria or research or educational conference grants from Lundbeck and Otsuka. Othman Sentissi has received advisory board honoraria or research or educational conference from Otsuka, Lilly, Lundbeck, Sandoz, Janssen and Sunovion on an institutional account for research and teaching. Prof. Gordon Parker has received lecture fees and board member honoraria from Otsuka, Servier and Lundbeck. Dr. David Bond has received consulting fees and/or research grants from Alkermes, Myriad Genetics, the National Institutes of Health, the University of Minnesota Department of Psychiatry and Behavioral Sciences, and the University of Minnesota Foundation. Prof. Giulio Perugi has received grant/research support from Eli Lilly & Co.; is on the speaker/advisory board of Sanofi-Aventis, Bristol-Myers Squibb, AstraZeneca, Eli Lilly & Co., Jannsen-Cilag, and Lundbeck; and has acted as consultant of AstraZeneca, Eli Lilly & Co., and Lundbeck. Prof. Eduard Vieta has received grants and served as consultant, advisor or CME speaker for the following entities: AB- Biotics, Abbott, Allergan, Angelini, Dainippon Sumitomo Pharma, Galenica, Janssen, Lundbeck, Novartis, Otsuka, Sage, Sanofi-Aventis, and Takeda. Dina Popovic has served as a speaker and/or medical writer and/or consultant and/or has participated in advisory boards for Bristol-Myers Squibb, Dexel, Merck Sharp & Dohme, Janssen-Cilag, Lundbeck, Ferrer, and Forum Pharmaceuticals. None of the remaining authors have conflicts of interest to declare., (Copyright © 2022 Elsevier B.V. and ECNP. All rights reserved.)

Background: The COVID-19 pandemic's mental health consequences remain unknown., Aim: To assess the mental health status of ambulatory cardiometabolic patients during COVID-19 pandemic lockdown in Spanish speaking Latin American countries., Methods: Cardiometabolic patients without COVID-19 evidence in 13 Latin American countries answered a survey between June 15th and July 15th, 2020. The Diagnosis Manual of Mental Disorders fifth edition was used to identify the presence of major depressive symptoms., Results: The sample included 4216 patients, 1590 (37.71%; IC95% 36.24-39.19) were considered suffering major depression. Female gender, consuming ≥5 medications day, physical activity <100 minutes weekly, low fruits and vegetables intake, poor treatment adherence, reduced food consumption were independently associated to the presence of major depressive symptoms., Conclusions: The CorCOVID Latam Psy study showed that one-third of the Latin American Spanish speaking population is suffering from major depressive symptoms during the COVID-19 outbreak., (Copyright © 2020. Published by Elsevier Inc.)

Background: The pentose phosphate pathway (PPP) has received significant attention because of the role of NADPH and R-5-P in the maintenance of cancer cells, which are necessary for the synthesis of fatty acids and contribute to uncontrollable proliferation. The HsG6PD enzyme is the rate-limiting step in the oxidative branch of the PPP, leading to an increase in the expression levels in tumor cells; therefore, the protein has been proposed as a target for the development of new molecules for use in cancer., Methods: Through in vitro studies, we assayed the effects of 55 chemical compounds against recombinant HsG6PD. Here, we present the kinetic characterization of four new HsG6PD inhibitors as well as their functional and structural effects on the protein. Furthermore, molecular docking was performed to determine the interaction of the best hits with HsG6PD., Results: Four compounds, JMM-2, CCM-4, CNZ-3, and CNZ-7, were capable of reducing HsG6PD activity and showed noncompetitive and uncompetitive inhibition. Moreover, experiments using circular dichroism and fluorescence spectroscopy showed that the molecules affect the structure (secondary and tertiary) of the protein as well as its thermal stability. Computational docking analysis revealed that the interaction of the compounds with the protein does not occur at the active site., Conclusions: We identified two new compounds (CNZ-3 and JMM-2) capable of inhibiting HsG6PD that, compared to other previously known HsG6PD inhibitors, showed different mechanisms of inhibition., General Significance: Screening of new inhibitors for HsG6PD with a future pharmacological approach for the study and treatment of cancer., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Aims: Haloperidol is a neuroleptic drug with high affinity towards the σ 1 receptor (σ 1 R), acting as antagonist that decreases neuropathic pain, but has CNS side effects. This work describes the design and synthesis of a novel analog N‑(1‑benzylpiperidin‑4-yl)‑4‑fluorobenzamide (LMH-2), which produced antihyperalgesic and antiallodynic effects in rats with neuropathy induced by chronic constriction injury of the sciatic nerve (CCI), being more active than gabapentin (The most widely used drug for the treatment of neuropathic pain)., Main Methods: LMH-2 was designed as haloperidol analog. Its structure was characterized by spectroscopic ( 1 H and 13 C NMR) and spectrometric mass (electronic impact) techniques. Additionally, in silico predictions of pharmacokinetic, pharmacodynamic and toxicological properties were obtained, with promising results. A competitive binding assay using radioligands was employed to evaluate the in vitro affinity for σ 1 R, whereas in vivo antihyperalgesic and antiallodynic activities were investigated using Wistar rats with CCI., Key Findings: LMH-2 showed high affinity for σ 1 R in an in vitro binding assay, with a K i  = 6.0 nM and a high σ 1 R/σ 2 R selectivity ratio. Molecular docking studies were carried out to determine the binding energy and to analyze LMH-2-protein interactions. Through an in silico pharmacological consensus analysis, LMH-2 was considered safe for in vivo evaluation. Thus, LMH-2 had dose-dependent antiallodynic and antihyperalgesic activities; its efficacy was comparable to that of gabapentin, but its potency was 2-times higher than this drug., Significance: LMH-2 administration produced antihyperalgesic and antiallodynic effects by the antagonism of σ 1 R, suggesting its potential use as an analgesic drug for neuropathic pain., Competing Interests: Declaration of competing interest The authors declare no conflict of interest., (Copyright © 2020 Elsevier Inc. All rights reserved.)

The fast development of today's healthcare and the need to extract new medical knowledge from exponentially-growing volumes of standardized Electronic Health Records data, as required by studies in Precision Medicine, brings up a challenge that may probably only be addressed using NoSQL DBMSs, due to the non-optimal performance of traditional relational DBMSs on standardized data; and these database systems operated by semantic archetype-based query languages, because of the expected generalized extension of standardized EHR systems. An AQL into MongoDB interpreter has been developed to its first version. It translates system-independent AQL queries posed on ISO/EN 13606 standardized EHR extracts into the NoSQL MongoDB query language. The new interpreter has the advantages of both the archetype-based system-independent AQL queries and the dual-model-based standardized EHR extracts stored on document-centric NoSQL DBMSs, such as MongoDB. AQL queries are independent of applications, programming languages and system environments due to the use of the dual model, but EHR extracts featuring this model are best persisted on document-based NoSQL databases. Consequently, the interpreter allows us to query standardized EHR extracts semantically, and also affording optimal performance., (Copyright © 2019 Elsevier Inc. All rights reserved.)

Aim: N-(2,6-Dichlorophenyl)-2-(4-methyl-1-piperidinyl)acetamide (LIA), a lidocaine analogue, has potential applications in treating neuropathic pain. The aim of this work was to characterize the pharmacological activity of LIA related with central nervous system and cardiovascular activity., Methods: Anesthetic effect was tested in guinea pigs and mice. Ambulatory activity, anti-anxiety effect, sodium pentobarbital (PB)-induced hypnosis and pentylenetetrazol (PTZ)-induced seizures test were evaluated in mice to determine the possible central nervous system activity. The cardiovascular activities in vivo and ex vivo were analyzed in rats., Key Findings: LIA (2%) presents, similar to lidocaine (2%), anesthetic activity on the corneal reflex, infiltration anesthesia and tail immersion test. LIA (1-100mg/kg, i.p.), similar to lidocaine (1-100mg/kg, i.p.), presents a dose-dependent sedative-hypnotic effect in mice. Both compounds did not produce anti-anxiety activity in mice. LIA did not prevent PTZ-induced seizures. However, LIA itself did not produce seizures at high doses in mice, as lidocaine does. LIA is a vasorelaxant compound for smooth muscle cells and presents hypotensive effect in vivo without increments to the heart rate significantly., Significance: High doses of lidocaine produce seizures and vasoconstriction. In this study, we found that LIA shares a similar pharmacological profile as lidocaine's but without the primary adverse effects of seizures and vasoconstriction., (Copyright © 2016 Elsevier Inc. All rights reserved.)

New and stimulating results have challenged the concept that cellular senescence might not be synonymous with aging. It is indisputable that during aging, senescent cell accumulation has an impact on organismal health. Nevertheless, senescent cells are now known to display physiological roles during embryonic development, during wound healing repair and as a cellular response to stress. The fact that senescence has been found in cells that did not attain their maximal round of replications, nor have metabolic alterations or DNA damage, also challenges the paradigm that senescence is cellular aging, and it is in favor of the idea that cellular senescence is a phenomenon that has a function by itself. Therefore, in order to understand this phenomenon it is important to analyze the relationship between senescence and other cellular responses that have many features in common, such as apoptosis, cancer and autophagy, particularly highlighting their role during development and adulthood., (Copyright © 2016 Elsevier Inc. All rights reserved.)

Estrogenic chemicals are often detected in the aquatic environment and can negatively affect animal development and reproduction. In teleost fishes, the hormonal regulation during a critical period of larval development has a strong influence on gonadal sex differentiation; thus this process may be affected by the exposure to environmental estrogens. In this study, we first assessed the lethal acute toxicity of the natural estrogen 17β-estradiol (E2) and the weaker estrogen mimics 4-tert-octylphenol (OP) and 4-nonylphenol (NP) on larval stages of the South American cichlid fish Cichlasoma dimerus. In a further experiment, we analyzed the effects of chronic waterborne exposure to E2 and OP on gonad development and sex differentiation. Exposure to high concentrations of E2 had a pronounced feminizing effect directing sex differentiation towards ovarian development, while testis development was inhibited at a lower, environmentally relevant concentration. Among OP-exposed fish, 15-38.5% of the males exhibited testicular oocytes (TOs), a commonly reported biomarker of estrogenic exposure. However, since TOs were also recorded in control males and the proportion of males with TOs was not significantly higher in OP treatments, their occurrence could not be attributed to OP exposure. In addition, TOs did not seem to impair male gonad development and functionality since normal spermatogenesis was observed in testes of OP-treated fish. These results indicate that E2 occurring in the South American aquatic environment may affect male reproductive development and pose a risk for wild C. dimerus, especially under prolonged exposure, while the effects of weaker xenoestrogens such as OP would be negligible for gonad development in this species. As illustrated by this study, the natural occurrence of TOs indicates that conclusions concerning the causes of this phenomenon must be drawn with care., (Copyright © 2015 Elsevier Inc. All rights reserved.)

In humans, 2 functionally different types of adipose tissue coexist: white adipose tissue (WAT) and brown adipose tissue (BAT). WAT is involved in energy storage, whereas BAT is involved in energy expenditure. Increased amounts of WAT may contribute to the development of metabolic disorders, such as obesity-associated type 2 diabetes mellitus and cardiovascular diseases. In contrast, the thermogenic function of BAT allows high consumption of fatty acids because of the activity of uncoupling protein 1 in the internal mitochondrial membrane. Interestingly, obesity reduction and insulin sensitization have been achieved by BAT activation-regeneration in animal models. This review describes the origin, function, and differentiation mechanisms of BAT to identify new therapeutic strategies for the treatment of metabolic disorders related to obesity. On the basis of the animal studies, novel approaches for BAT regeneration combining stem cells from the adipose tissue with active components, such as melatonin, may have potential for the treatment of metabolic disorders in humans., (Copyright © 2015 Elsevier Inc. All rights reserved.)

Unlabelled: Pilot study to validate a Computerized Decision Support Systems (CDS) (HTE-DLP) for improving treatment of hyperlipidemia., Methods: HTE-DLP was programmed to offer automatic specific reminders for lipid treatment. Seventy-seven patients with high cardiovascular risk were randomized. 10 expert physicians in cardiovascular-risk management were recruited. We assessed number of patients at LDL <70 mg/dl after 12 weeks of treatment., Results: A greater proportion of intervention group reached the LDL-C <70 mg/ml [55.0% vs 12.5%, p = 0.003; OR: 3.26 IC (1.16-9.15)]. "High potency statins" and combined therapy were used more frequently in the intervention group than the control group (p = 0.001). Seven adverse effects were documented in the intervention group and two in the control group. An acceptable relationship was observed with regard to costeffectiveness in the intervention group. Physicians expressed good agreement with HTE-DLP (86.1%) and comfortable ease-of-use (85%)., Conclusions: Use of a CDSS in high-risk cardiovascular patients resulted in a significant reduction in LDL-C levels., (Copyright © 2013 Elsevier Ireland Ltd. All rights reserved.)

Aims: To investigate the local antinociceptive effect as well as the possible mechanisms of action of a novel analogue of palmitoylethanolamide (PEA) N-(4-methoxy-2-nitrophenyl)hexadecanamide (HD) in the rat formalin test., Main Methods: The formalin test was used to assess the antinociceptive activity of HD in vivo. The hydrolysis of anandamide catalyzed by fatty acid amide hydrolase (FAAH) was used to determine the action of HD on FAAH activity in vitro., Key Findings: Local peripheral ipisilateral, but not contralateral, administration of HD (10-100μg/paw) produced a dose-dependent antinociceptive effect in rats. The CB(1) and CB(2) receptor antagonists AM281 (0.3-30μg/paw) and SR144528 (0.3-30μg/paw), respectively, reduced the antinociceptive effect of HD (100μg/paw). In addition, methiothepin (0.03-0.3μg/paw) and naloxone (5-50μg/paw) significantly reduced HD-induced antinociception (100μg/paw). In vitro, HD reduced only to a minor extent the hydrolysis of anandamide catalyzed by FAAH., Significance: HD local administration produces antinociception that probably results from an indirect activation of peripheral CB(1) and CB(2) cannabinoid receptors. Data suggest that 5-HT(1) and opioid receptors also participate in the antinociceptive effect of this compound. HD may have potential as analgesic drug., (Copyright © 2012 Elsevier Inc. All rights reserved.)

The organochlorine insecticide endosulfan (ES) is widely used despite its high toxicity to fish (96-h LC(50) median value of 2.6 μg L(-1)). This study aimed to assess the acute toxicity, histological and physiological parameters after exposure to 0; 0.25; 1; 2; 3; 4 and 16 μg L(-1) ES for 96 h under semi-static conditions in a freshwater perciform fish, Cichlasoma dimerus. Prior to death, fish exhibited behavior indicative of neurotoxicity. No difference was found in brain AChE activity. A decrease in erythrocyte mean corpuscular volume, mean corpuscular hemoglobin and mean corpuscular hemoglobin concentration in exposed fish suggests a state of anemia. Histological alterations observed in exposed fish included hyperplasia of the interlamellar epithelium, blood congestion in secondary lamellae, and mucous cells hyperplasia and hypertrophy in gills; pycnotic nuclei and hydropic degeneration in liver; testicular damage. These moderate pathological responses in major organs could become crucial during reproduction and under prolonged exposure periods., (Copyright © 2011 Elsevier Inc. All rights reserved.)

The influence of initial cation concentration, temperature and pH was investigated to optimize Pb(2+), Cu(2+) and Zn(2+) removal from aqueous solutions using acid formaldehyde pre-treated chestnut shell as adsorbent. Experiments were planned according to an incomplete 3(3) factorial experimental design. Under the optimal conditions selected, the metal ion adsorption equilibrium was satisfactorily described by the Langmuir isotherm model. The maximum pre-treated chestnut shell adsorption capacity was obtained for Pb(2+) ions, 8.5 mg g(-1), and the order of cation affinity was Pb(2+)>Cu(2+)>Zn(2+). A model that considered the effect of axial dispersion was successfully used to describe the fixed-bed adsorption behaviour of Pb(2+), Cu(2+) and Zn(2+) ions at the flow rates essayed. Fourier transform infrared (FTIR) and X-ray photoelectron (XPS) spectroscopies showed that the functional groups involved in metal ions binding included carboxyl, hydroxyl, ether, alcoholic and amino groups.

Protein-RNA interactions are involved in all facets of RNA biology. The identification of small molecules that selectively block such bimolecular interactions could provide insight into previously unexplored steps of gene regulation. Such is the case for regulation of eukaryotic protein synthesis where interactions between messenger RNA (mRNA) and several eukaryotic initiation factors govern the recruitment of 40S ribosomes (and associated factors) to mRNA templates during the initiation phase. We have designed simple fluorescence polarization-based high-throughput screening assays that query the binding of several translation factors to RNA and found that the mixed inhibitor p-chloromercuribenzoate interferes with poly(A) binding protein-RNA interaction.

Hot aqueous solutions of a wide variety of aliphatic amines bring about efficient cleavage of 5'-(32)P-labeled oligodeoxyribonucleotides. Electrophoretic resolution of the product mixtures produces a radioactivity pattern that can be analyzed in terms of relative band intensities and band separations to deduce the base sequence of the original oligonucleotide. Amines differ in their overall reactivity with respect to the oligonucleotide as well as in their preference for reaction with the various heterocyclic bases. Guanine sites, in particular, vary markedly in their cleavage susceptibility when reacted with the different amines, being more vulnerable to scission when reacted with less basic amines. Guanine cleavage propensity can also be affected markedly by the ionic strength of the aminolysis solution. Both effects are interpreted in terms of variable extents of deprotonation of the guanine sites in the basic medium. Some of the amines produce fragment patterns that are marred by the presence of minor extraneous bands; these probably are due to an incomplete extent of the second beta-elimination involved in backbone cleavage. The methodology is applicable to confirmatory sequencing of synthetic oligonucleotides and can also be used to prepare standard ladders for use in footprinting experiments or chemical reactivity studies.

We have determined that the methanolic extract of L. caulescens (MELc) produced a significant vasodilator effect in a concentration-dependent and endothelium-dependent manner. This relaxation was blocked by N(omega)-nitro-L-arginine methylester (L-NAME), indicating that MELc vasodilator properties are endothelium mediated due to liberation of nitric oxide (NO). In this paper we aimed to corroborate its mode of action. MELc effects on noradrenaline (NA)-induced contraction in isolated rat aortic thoracic rings with endothelium (+E), in the presence of atropine (0.1 microM) and 1-H-[1,2,4]-oxadiazolo-[4,3a]-quinoxalin-1-one (ODQ, 1 microM) were conducted. MELc relaxation curve was significantly shifted to the right in the presence of ODQ and atropine, thus confirming that its mode of action is related with activation of nitric oxide synthase (NOS) and the consequent increment in NO formation. Bio-guided study of MELc allowed the isolation of ursolic acid (UA, 50 mg) and ursolic-oleanolic acids mixture [UA/OA (7:3), 450 mg]. The relaxant effect of UA (0.038-110 microM) was evaluated in functional experiments. UA induced a significant relaxation in a concentration- and endothelium-dependent manner (IC(50)=44.15 microM) and did not produce a vasorelaxant effect on contraction evoked by KCl (80 mM). In addition, NA-induced contraction was significantly displaced to the right by UA (30 microM). In order to determine its mode of action, UA-induced relaxant effect was evaluated in the presence of atropine (0.1 microM), indomethacin (10 microM), L-NAME (100 microM) and ODQ (1 microM). Relaxation was blocked by L-NAME and ODQ. On the other hand, UA (3 microM) provoked a significant displacement to the left in the relaxation curve induced by sodium nitroprusside (SNP, 0.32 nM to 0.1 microM), but it was not significant in the presence of Carbamoyl choline (carbachol, 1 nM to 10 microM). These results indicate that UA-mediated relaxation is endothelium dependent, probably due to NO release, and the consequent activation of vascular smooth muscle soluble guanylate cyclase (sGC), a signal transduction enzyme that forms the second messenger cGMP.

The relaxant activity of 2-(o, p-substituted phenyl)-1H-benzimidazole derivatives with various 5- and 6-position substituents (-H, -CH3, -NO2, -CF3), namely 1-7, was recorded using the in vitro rat aorta ring test. Compounds 3 and 6 [2-(5-nitro-1H-benzimidazol-2-yl)phenol and 2-(4-methoxyphenyl)-5-nitro-1H-benzimidazole] were prepared using a short route, and were the most potent compounds of the series, showing IC50 value of 0.95 and 1.41 (with endothelium) and 2.01 and 3.61 microM (without endothelium), respectively. Studying further structure-activity relationships through the use of bioisosteric substitution in these benzimidazole derivatives should provide novel vasorelaxant leads and possibly against hypertensive diseases.

The adsorption of phenol from aqueous solutions using a column packed with pre-treated Pinus pinaster bark was studied. The influence of the inlet phenol concentration (0.01 or 0.1 g/L) and the flow rate (6, 15.6 or 30 mL/min) on the breakthrough curves was analysed. An increase in the flow rate, decreased the time necessary to reach the breakthrough point and, for the highest inlet concentration, the dynamic capacity of the bed, from 7.5 to 0.4 min and from 0.38 to 0.15 mg phenol/go.d. bark, respectively, at 0.1 g/L. The LUB Design Approach was used to determine the equivalent length of unused bed. The lower LUB values, which imply a better utilization of the bark bed, were obtained at the higher flow rate. A model which considered the effect of axial dispersion was successfully used to describe the fixed-bed operation behaviour for the lower flow rates. For the lowest inlet phenol concentration, the axial dispersion coefficient increased significantly when the flow rate increased.