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Purpose: The 3-category rating of volar plate prominence in relation to the most volar edge of the distal radius (the watershed line) on lateral radiographs was reliable among a small group of surgeons and associated with the probability of flexor tendon irritation and potential rupture. Classifications are often less reliable when tested among a large group of practicing surgeons in different environments. Methods: In this survey-based experiment, an international group of 115 fracture and upper extremity surgeons viewed 1 of 4 sets of 24 lateral radiographs (96 unique lateral radiographs) of patients with distal radius fractures who underwent volar plating in the practice of a single surgeon using 2 types of plates. Surgeons were asked to rate the following metrics: (1) the grade of plate prominence according to Soong, (2) whether the plate was more prominent than the watershed line, (3) whether the plate was separate from the bone distally, and (4) whether there is more than 5° of dorsal angulation of the distal radius articular surface. Results: The interobserver agreement of the classification was “fair” (κ = 0.32; 95% confidence interval [CI] = 0.27–0.36), and grading was more reliable among surgeons who do not supervise trainees. Volar prominence was less reliable (κ = 0.034; 95% CI = 0.013–0.055) than plate separation from bone (κ = 0.50; 95% CI = 0.42–0.59) and more than 5° of dorsal angulation (κ = 0.42; 95% CI = 0.35–0.48). Conclusions: Among a large number of international practicing surgeons, the classification of volar plate prominence in 3 categories was fair. Clinical Relevance: The diagnosis of plate prominence might develop toward criteria with moderate reliability, such as separation of the plate from the bone and residual angulation of the distal radius.

Composites are materials formed by the combination of two or more components that acquire better properties than the ones obtained by each component on its own. Composites have been widely used in the industry due to its light weight and good mechanical properties. To improve these properties several layers of reinforced material (e.g., carbon fibre) are overlapped which produce an increase in the fibre consumption. In this sense Tailored Fibre Placement (TFP) embroidery can offer good opportunity to reduce the consumption of reinforced fibre while improving the mechanical properties due to the alignment of the fibres in the effort direction. This study analyzes the performance of carbon fibre reinforced composites with Polyester resin made with TFP embroidery technology against flexural strength efforts and without using plain woven fabrics to demonstrate that the use of reinforcement fabrics in composites can be optimized by a curved alignment of the fibers. Two different structures were embroidered with TFP technology, one simulating a woven fabric with straight unidirectional alignment of fibres in horizontal and vertical direction, and a second structure made with curvilinear alignment of carbon fibers. After the study of the flexural mechanical properties an improvement of 18% was obtained in maximum flexural strength.

Aims: The aim of this study is to analyze the impact of 10 years of blended learning in echocardiography. Methods and results: A total of 140 of 210 medical doctors who graduated from our teaching program from 2011 to 2020 responded to the survey. Among them, 53,57% were anesthesiologists and 26,42% intensivists. More than 85% of respondents indicated that the online period met their expectations, and 70,2% that the hands on practice met the objectives. Posterior to the Diploma there was a significant increase in the frequency of use of echo compared to before the Diploma, both in transthoracic and transesophageal echocardiography, from 24,29% to 40,71% and from 13,57% to 27,86%, respectively. The echo was applied mainly in the perioperative period (56,7%) and in intensive care (32,3%), and only in 11% of cases it was used in emergency units. Additionally, 92,4% of respondents revealed that it was very useful in their subsequent professional practice and 92,47% that it helped them in the interpretation of echo reports. Among the strengths they highlighted ‘the theoretical content, clinical cases, hands on activity and the teaching staff’. Among the weaknesses, 50% requested a practice period lasting more than one week. Conclusion: The blended learning of echocardiography over a decade has been satisfactorily evaluated by the graduated specialists and has been related to a significant increase in its use in the perioperative period and in intensive care. The main challenges are to establish a longer period of practice and achieve greater scope in Emergency Medicine.

To assess skull bone thickness from birth to skeletal maturity at different sites to provide a reference for the correct selection of pin type and pin placement according to age.270 children and adolescents (age: 0-17 years) with a normal CT scan obtained at Emergency Department for other medical reasons were included. Skull thickness was measured on the axial plane CT scans at eight different sites of the vault: midline anterior (A) and posterior (P), right and left lateral (L), antero-lateral (AL), postero-lateral (PL).From birth to skeletal maturity, L thickness was increased significantly less (+ 58%) compared with AL (+ 205%), P (+ 233%), PL (+ 247%), and A (+ 269%) thickness (P 0.01). At the end of growth, the thickest and thinnest points of the vault (absolute value) were found at the P and L measurement sites, respectively (P 0.01). Children aged 4 years exhibited the highest variability in AL and PL skull bone thickness, with thickness 3 mm observed in 85% (64/75 patients) and 92% (69/75 patients) of cases, respectively.We recommend that the tip of the pin should not exceed 2-3 mm in children aged 4, and 4 mm in children aged 4-6 years, to decrease the risk of inner table perforation. After the age of 7 years and 13 years, standard-sized pin tips (5 and 6 mm, respectively) may be safely used. Children aged 4 years show significant variability in skull thickness, and therefore a CT scan may be required for this particular age group.

In recent years, there has been a trend of increasing criticism towards official development assistance (ODA) and the optimization of ODA policies, in a world of growing inequality between the Global North and Global South. To contribute to efficient ODA planning, this article proposes to innovatively apply system dynamics as an optimal tool for ex ante impact assessment. The study case is located in the slums of Darkhan (Mongolia), whose citizens and environment suffer the consequences of poor urban planning and lack of municipal solid waste management (MSWM). In this context, the present research proposes a policy of education and infrastructure as key factors for the improvement of MSWM in the context of an international cooperation plan, carried out by the Korean agency KOICA. To evaluate its impact and anticipate its effects, a tailor-made system dynamics model of a Darkhan district has been created, with the focus on the education process in order to simulate the different options of the proposed policies. The results show that education policy is particularly relevant for behavioral change, in terms of reducing waste burned and waste on the ground, and increasing composted and recycled waste. However, in this context, the policy is ineffective for improving the district’s water and air pollution situation. This article also offers discussions and recommendations to be applied to the international cooperation plan, which takes place in real life. It is expected that the described process of model construction and its results will contribute to the further use of system dynamics as a planning tool in the international cooperation field.

Statistical inference was introduced by Fisher and Neyman-Pearson more than 90 years ago to define the probability that the difference in results between several groups is due to randomness or is a real, “significant” difference. The usual procedure is to test the probability (P) against the null hypothesis that there is no real difference except because of the inevitable sampling variability. If this probability is high we accept the null hypothesis and infer that there is no real difference, but if P is low (P < 0.05) we reject the null hypothesis and infer that there is, a “significant” difference. However, a large amount of discoveries using this method are not reproducible. Statisticians have defined the deficiencies of the method and warned the researchers that P is a very unreliable measure. Two uncertainties of the “significance” concept are described in this review: a) The inefficacy of a P value to discard the null hypothesis; b) The low probability to reproduce a P value after an exact replication of the experiment. Due to the discredit of “significance” the American Statistical Association recently stated that P values do not provide a good measure of evidence for a hypothesis. Statisticians recommend to never use the word “significant” because it is misleading. Instead, the exact P value should be stated along with the effect size and confidence intervals. Nothing greater than P = 0.001 should be considered as a demonstration that something was discovered. Currently, several alternatives are being studied to replace the classical concepts.

Statistical inference was introduced by Fisher and Neyman-Pearson more than 90 years ago to define the probability that the difference in results between several groups is due to randomness or is a real, "significant" difference. The usual procedure is to test the probability (P) against the null hypothesis that there is no real difference except because of the inevitable sampling variability. If this probability is high we accept the null hypothesis and infer that there is no real difference, but if P is low (P0.05) we reject the null hypothesis and infer that there is, a "significant" difference. However, a large amount of discoveries using this method are not reproducible. Statisticians have defined the deficiencies of the method and warned the researchers that P is a very unreliable measure. Two uncertainties of the "significance" concept are described in this review: a) The inefficacy of a P value to discard the null hypothesis; b) The low probability to reproduce a P value after an exact replication of the experiment. Due to the discredit of "significance" the American Statistical Association recently stated that P values do not provide a good measure of evidence for a hypothesis. Statisticians recommend to never use the word "significant" because it is misleading. Instead, the exact P value should be stated along with the effect size and confidence intervals. Nothing greater than P = 0.001 should be considered as a demonstration that something was discovered. Currently, several alternatives are being studied to replace the classical concepts.

The concept of the “mesenchymal stromal cell” (MSC) has emerged following a number of studies starting from the middle of the 19th century that led to the discovery of the intrinsic osteogenic potential of bone marrow (BM) cells and their capacity to support hematopoiesis. Thus, a relationship was established between bone formation and the development of hematopoiesis. A mesenchymal precursor cell was identified in BM that displays differentiation potential towards osteocytic, chondrocytic, and adipocytic lineages and hematopoiesis-supporting capacity. Currently, “mesenchymal stromal cell” is the most commonly accepted term for this cell type. Thereafter, the concept of “hematopoietic stem cell (HSC) niche” was developed. This is a specific place where BM stromal cells are in intimate contact with HSCs to control their behavior. Very recently published data have helped to better characterize in vivo the cellular composition and anatomical localization of the HSC niche. Initially described in BM, MSCs have been found in almost all pre- and postnatal tissues. The actual interest for MSCs is mainly due to their multipotency ability with and thus their potential use for tissue repair, as well as their contribution to tumoral niches that could represent new antitumoral therapy targets.

TERMINAL FLOWER 1 (TFL1) is a key regulator of Arabidopsis plant architecture that responds to developmental and environmental signals to control flowering time and the fate of shoot meristems. TFL1 expression is dynamic, being found in all shoot meristems, but not in floral meristems, with the level and distribution changing throughout development. Using a variety of experimental approaches we have analysed the TFL1 promoter to elucidate its functional structure. TFL1 expression is based on distinct cis-regulatory regions, the most important being located 3' of the coding sequence. Our results indicate that TFL1 expression in the shoot apical versus lateral inflorescence meristems is controlled through distinct cis-regulatory elements, suggesting that different signals control expression in these meristem types. Moreover, we identified a cis-regulatory region necessary for TFL1 expression in the vegetative shoot and required for a wild-type flowering time, supporting that TFL1 expression in the vegetative meristem controls flowering time. Our study provides a model for the functional organisation of TFL1 cis-regulatory regions, contributing to our understanding of how developmental pathways are integrated at the genomic level of a key regulator to control plant architecture., This work was supported by a Joint Project Grant from the Royal Society [ESEP/JP 15057] to D.B. and F.M. The laboratory of F.M. was funded by grants from the Spanish Ministerio de Ciencia e Innovacion [BIO2009-10876 and CSD2007-00057], the Spanish Ministerio de Economia y Competitividad [BFU2012-38929] and from the Generalitat Valenciana [ACOMP09-083 and ACOMP2012-099]. Work in the Y.H. lab was supported by the Plant Genome Research Program from the National Science Foundation [NSF-PGRP-IOS-1339388]. P.F.-N. was supported by a fellowship from the I3P Program of Consejo Superior de Investigaciones Cientificas.

The contractile state of the heart is the result of myocardial contractility, the intrinsic mechanism that regulates the force and the shortening of the ventricle and determines the ventricular ejection volume. However, the ejection volume is also modulated by ventricular preload (diastolic ventricular volume) and afterload (resistance to ejection). Accordingly, a decrease in contractility may be masked by changes in preload or afterload, maintaining a normal ejection volume and delaying the diagnosis of myocardial damage. Thus, it is necessary to develop a non-invasive method to measure contractility in the clinical practice. We review in this article the basic principles of cardiac contraction, the concept of contractility and its measurement with the ventricular pressure-volume loop, an experimental method that also measures most of the hemodynamic variables of the cardiac cycle including preload, afterload, ventricular work, ventricular lusitropy and arterial elastance. This method has been recently validated in cardiac patients and allows to evaluate the evolution of contractility in heart failure in a non invasive way. Although some modifications are still necessary, it will probably have an extensive use in practical cardiology in the near future.

A study on the transdermal permeation through human skin was performed with a series of 6 semisolid formulations (A-F) containing 1% sodium diclofenac (CAS 15307-79-6) (w/w). A commercially available drug preparation was tested as a reference. Based on permeation characteristics, a study on the topical and systemic anti-inflammatory activities of three formulations (A, F and the reference formulation) was conducted using the model of erythema induced by UV radiation in hairless rats. This is expected, together with the index of topical anti-inflammatory activity to allow the selection of the most suitable formulation for dermal application. The following representative parameters were measured in the permeation study: amount of diclofenac permeated at 24 h, flow, lag time and amount of drug retained in skin at 24 h. Of the formulations tested, diclofenac formulated in the reference formulation showed the highest values of amount of diclofenac permeated at 24 h, amount of drug retained in skin at 24 and flow. As regards the skin inflammation test, no significant differences (p 1, it is concluded that a good therapeutic performance could be obtained in the treatment of local inflammation with diclofenac by using formulation F.

We explored the influence of polymorphisms in genes encoding the chemokine stromal cell–derived factor-1 (SDF-1)/CXCL12 in a cohort of Tunisian patients with malignant hematologic diseases multiple myeloma [MM], non-Hodgkin's lymphoma [NHL], Hodgkin's disease, and acute myeloid leukemia [AML], who underwent stem cell mobilization for autologous transplantation versus a group of healthy donors for allogeneic transplantation. Polymerase chain reaction–restriction fragment length polymorphism (PCR-RFLp) analysis was used for rapid identification of genotypes. Significant associations for SDF1-3′A polymorphism were observed exclusively in patients with MM and NHL. While there was a lack of all association of SDF-1 polymorphism with AML patients. However, considering that the ability of mobilization varies among subjects, we have observed that the SDF1-3′A allele was associated with good mobilization capacity. Interestingly, the association was mainly observed among healthy allogeneic transplant donors where the analysis was not biased by background disease or chemotherapy (P = .010; odds ratio = 2.603; confidence interval [95%] = 1.239–5.466).

Fading immune protection in farmed animals may present a problem, particularly in free-ranging animals in nomadic and transhumant pastoral systems, where animals are not readily available for large-scale blanket vaccination programmes. Two veterinary examples of fading immune protection are discussed: rinderpest and foot-and-mouth disease (FMD). Both are devastating viral diseases of cattle that have a huge impact on the farming economy. Both diseases can be controlled by vaccination, although the post-vaccination immunity afforded by the rinderpest vaccine is markedly different from that induced by FMD vaccines. These differences may in part explain the respective advancement of international eradication campaigns: while global eradication of rinderpest is imminent, FMD viruses are still actively circulating in many parts of the world.

The halophilic enzyme D-2-hydroxyacid dehydrogenase (D2-HDH) from Haloferax mediterranei is found to be an oligomeric enzyme composed of two identical subunits. Fluorescence spectra of native and denatured protein and effect of denaturants such as urea and guanidine hydrochloride on enzyme activity of halophilic D-2-hydroxyacid dehydrogenase have been analysed. Native D2-HDH shows the maximum emission at 340 nm. The denaturation process caused an exposure to the solvent of the tryptophan residues, as manifested by the red shift of the emission maximum from 340 to 350 nm. When urea was used as denaturant agent the enzyme required long incubation times, higher to 24 h, to unfold. Fluorescence quenching by KI and acrylamide was also carried out; showing that the tryptophan residues are mainly located near the enzyme surface.

Highlight Plants carefully control where and when flowers are made through activators and repressors. We show that spatially the shoot meristem is key in responding to the repressors of flowering TFL1., Models for the control of above-ground plant architectures show how meristems can be programmed to be either shoots or flowers. Molecular, genetic, transgenic, and mathematical studies have greatly refined these models, suggesting that the phase of the shoot reflects different genes contributing to its repression of flowering, its vegetativeness (‘veg’), before activators promote flower development. Key elements of how the repressor of flowering and shoot meristem gene TFL1 acts have now been tested, by changing its spatiotemporal pattern. It is shown that TFL1 can act outside of its normal expression domain in leaf primordia or floral meristems to repress flower identity. These data show how the timing and spatial pattern of TFL1 expression affect overall plant architecture. This reveals that the underlying pattern of TFL1 interactors is complex and that they may be spatially more widespread than TFL1 itself, which is confined to shoots. However, the data show that while TFL1 and floral genes can both act and compete in the same meristem, it appears that the main shoot meristem is more sensitive to TFL1 rather than floral genes. This spatial analysis therefore reveals how a difference in response helps maintain the ‘veg’ state of the shoot meristem.

The contractile state of the heart is the result of myocardial contractility, the intrinsic mechanism that regulates the force and the shortening of the ventricle and determines the ventricular ejection volume. However, the ejection volume is also modulated by ventricular preload (diastolic ventricular volume) and afterload (resistance to ejection). Accordingly, a decrease in contractility may be masked by changes in preload or afterload, maintaining a normal ejection volume and delaying the diagnosis of myocardial damage. Thus, it is necessary to develop a non-invasive method to measure contractility in the clinical practice. We review in this article the basic principles of cardiac contraction, the concept of contractility and its measurement with the ventricular pressure-volume loop, an experimental method that also measures most of the hemodynamic variables of the cardiac cycle including preload, afterload, ventricular work, ventricular lusitropy and arterial elastance. This method has been recently validated in cardiac patients and allows to evaluate the evolution of contractility in heart failure in a non invasive way. Although some modifications are still necessary, it will probably have an extensive use in practical cardiology in the near future.

Hematologic diseases are a significant part of health disorders in Benin. As an example, anemia is the second cause of hospitalization, measuring up to 7.9% all over the country (National Plan of Sanitary Development, 2009-2018). By contrast, there is only one active hematologist in the country. Thanks to two partnerships, on one hand between the health sciences faculty in Cotonou (Benin) and the medicine one in Tours (France), and on the other hand between the Beninese Blood Transfusion National Agency and the French Blood Establishment, a first blood transfusion and hematology formation was held in Cotonou on December 2014. Among other benefits, was created an hematology-transfusion network in order to facilitate relations between Beninese hospital doctors, with the support of the two French partner institutions. The article describes this progress.

We showed that exercise induces early and late myocardial preconditioning in dogs and that these effects are mediated through nicotinamide adenine dinucleotide phosphate reduced form (NADPH) oxidase activation. As the intracoronary administration of calcium induces preconditioning and exercise enhances the calcium inflow to the cell, we studied if this effect of exercise triggers exercise preconditioning independently of its hemodynamic effects. We analyzed in 81 dogs the effect of blocking sarcolemmal L-type Ca channels with a low dose of verapamil on early and late preconditioning by exercise, and in other 50 dogs, we studied the effect of verapamil on NADPH oxidase activation in early exercise preconditioning. Exercise reduced myocardial infarct size by 76% and 52% (early and late windows respectively; P < 0.001 both), and these effects were abolished by a single low dose of verapamil given before exercise. This dose of verapamil did not modify the effect of exercise on metabolic and hemodynamic parameters. In addition, verapamil blocked the activation of NADPH oxidase during early preconditioning. The protective effect of exercise preconditioning on myocardial infarct size is triggered, at least in part, by calcium inflow increase to the cell during exercise and, during the early window, is mediated by NADPH oxidase activation.

International audience; In the field of oncology, research is now focused on the development of theranostic nanosystems that combine the functions of drug delivery and imaging for diagnosis/monitoring. In this context, we designed polyethylene glycol (PEG)ylated superparamagnetic iron oxide nanoparticles (SPIONs) for the delivery of doxorubicin (DOX), an antineoplastic agent. These DOX-loaded PEGylated SPIONs, or DLPS, should be useful for the delivery of DOX in vivo, as well as for magnetic drug targeting (MDT) and magnetic resonance imaging (MRI). The aim of this study was to evaluate the potential applications of DLPS in vivo as drug carrier systems for the reduction of xenograft breast tumors induced in nude mice. Prior to the animal model experiments, the main internalization pathways for the nanovectors in MDA-MB435 breast cancer cells were determined to be based on caveolae- and clathrin-mediated endocytosis. The time- and quantity-dependence of the nanoparticle uptake by the cells altered the in vitro cytotoxicity of the DLPS. The in vitro antiproliferative effect of the DLPS was dependent not only on DOX concentration, but also on the efficacy of nanoparticle internalization. Evaluation of the effect of DLPS treatment on xenograft tumors in nude mice showed that DLPS limited tumor growth in a manner comparable to that of free DOX under normal conditions of tumor growth. The application of an external magnetic field on tumors, i.e., MDT, did not improve the efficacy of the DLPS treatment. Nevertheless, the vectorization of DOX with DLPS appears to limit the hematologic side effects usually associated with DOX treatment.

The ability of six putative plant growth promoting rhizobacteria, isolated from the rhizosphere of Nicotiana glauca L., to stimulate growth and induce systemic resistance against Xanthomonas campestris CECT 95 in Arabidopsis thaliana L. Col-0 was evaluated. The six bacterial strains significantly reduced the disease symptoms caused by the pathogen compared to the controls, with the best results obtained with the Bacillus strain N11.37 and the Stenotrophomonas strain N6.8. These two strains were tested on A. thaliana NahG plants and jar1-1 and etr1-1 mutants, to elucidate whether the salicylic acid (SA)-dependent or SA-independent pathway was involved in the induction of systemic resistance. The results indicate that N6.8 induces the SA-dependent pathway. For N11.37 it is as yet not clear as in the etr1-1 mutants and NahG plants ISR is not expressed, while in jar1-1 it is. In addition, levels of SA were measured in Col-0 plants treated with N6.8 and N11.37 to confirm whether or not the two strains produced an increased level of SA. N6.8- and N11.37-induced plants showed higher levels of SA than the controls. It is concluded that N6.8 induces a SA-dependent pathway while N11.37 induces a pathway that is both ethylene (ET)- and SA-dependent.

A gene encoding a new d -2-hydroxyacid dehydrogenase (E.C. 1.1.1.) from the halophilic Archaeon Haloferax mediterranei has been sequenced, cloned and expressed in Escherichia coli cells with the inducible expression plasmid pET3a. The nucleotide sequence analysis showed an open reading frame of 927 bp which encodes a 308 amino acid protein. Multiple amino acid sequence alignments of the D-2-hydroxyacid dehydrogenase from H. mediterranei showed high homology with D-2-hydroxyacid dehydrogenases from different organisms and other enzymes of this family. Analysis of the amino acid sequence showed catalytic residues conserved in hydroxyacid dehydrogenases with d -stereospecificity. In the reductive reaction, the enzyme showed broad substrate specificity, although α-ketoisoleucine was the most favourable of all α-ketocarboxylic acids tested. Kinetic data revealed that this new D-2-hydroxyacid dehydrogenase from H. mediterranei exhibits dual coenzyme-specificity, using both NADPH and NADH as coenzymes. To date, all D-2-hydroxyacid dehydrogenases have been found to be NADH-dependent. Here, we report the first example of a D-2-hydroxyacid dehydrogenase with dual coenzyme-specificity.

We developed novel acrylic-based polymers that can be used as mucoadhesive delivery systems. Poly(acrylic acid) hydrogels were modified by grafting adhesion promoter chains such as poly(ethylene glycol) (PEG) onto their back-bone chains, thus promoting the adhesive process by interpenetration. The copolymers synthesized were designated as P(AA-g-EG). Hydrogels were synthesized using PEG of two different molecular weights, 1000 and 2000, and with varying molar feed ratio of AA-EG (20:80, 40:60, 60:40, 80:20, 12:88, 25:75, 44:56, 67:33). The copolymers were synthesized by using free radical solution UV-polymerization. The effects of different PEG-tethered structures on mucoadhesion were studied using a tensiometric testing and the work of adhesion was calculated. Preswollen P(AA-g-EG) copolymer films composed of 40% acrylic acid (AA) and 60% ethylene glycol (EG), containing PEG 1000 tethers, exhibited the highest value for the work of mucoadhesion, 130 x 10(-3)+/-27 x 10(-3) mJ, that is five times higher than the formulation composed of pure PAA. Based on these results and associated molecular analysis, we conclude that the higher mucoadhesive properties of this specific copolymer were the result of the synergistic effects of both monomers. AA functional groups allowed the polymer to form multiple hydrogen bonds with the glycoproteins present in the mucus. PEG tethers possibly acted as mucoadhesive promoters, enhancing interpenetration of polymer chains into the mucus.

We have shown previously that electrically induced tachycardia effectively produces myocardial preconditioning. Among other effects, tachycardia increases calcium release rates in microsomal fractions enriched in sarcoplasmic reticulum (SR) isolated from dog cardiac ventricular muscle. Here, we report that preconditioning tachycardia increased twofold the NADPH oxidase activity of isolated SR-enriched microsomal fractions, measured as NADPH-dependent generation of superoxide anion and hydrogen peroxide. Tachycardia also augmented the association of rac1 and the NADPH oxidase cytosolic subunit p47 phox to the microsomal fraction, without modifying the content of the membrane integral subunit gp91 phox . Microsomes from control animals displayed endogenous S -glutathionylation of cardiac ryanodine receptors (RyR2); in microsomal fractions isolated after tachycardia RyR2 S -glutathionylation levels were 1.7-fold higher than in controls. Parallel in vitro experiments showed that NADPH produced a transient increase in calcium release rates and enhanced 1.6-fold RyR2 S -glutathionylation in control microsomes but had marginal or no effects on microsomes isolated after tachycardia. Catalase plus superoxide dismutase, and the NADPH oxidase inhibitors apocynin and diphenyleneiodonium prevented the in vitro stimulation of calcium release rates and RyR2 S -glutathionylation induced by NADPH, suggesting NADPH oxidase involvement. Conversely, addition of reducing agents to vesicles incubated with NADPH markedly inhibited calcium release and prevented RyR2 S -glutathionylation. We propose that tachycardia stimulates NADPH oxidase activity, which by enhancing RyR2 redox modifications such as S -glutathionylation, would contribute to sustain faster calcium release rates during conditions of increased cardiac activity. This response may be an important component of tachycardia-induced preconditioning.

To estimate the applicability of transdermal drug permeation parameters in a finite-dose model for skin pre-treated with terpenes and to evaluate the enhancing effect of some terpene formulations on alprazolam permeation.In vitro enhancement of alprazolam human skin permeation was investigated using a pretreatment with different terpene solutions. Vertical diffusion, Franz-type cells were used. Intrinsic drug permeation was also investigated. Transdermal permeation parameters were estimated from the permeation tabulates using different theoretical approaches for their calculation. Two groups of permeation parameters were calculated: modelistic (diffusion of a finite-dose of drug model) and parameters nondependent of a diffusional model.In control experiments, all approaches of data treatment satisfactorily described the experimental permeation profiles. When skin pre-treatment was investigated, the fitting of a mathematical sigmoid function was much better than the diffusional approach. Pre-treatment of the skin with Limonene dissolved in ethanol / propylene glycol and Menthol dissolved in propylene glycol increased 15 and 10 times respectively the permeation parameters of alprazolam.Using enhancers that are rapidly cleared from the skin, skin permeability does not remain constant during the permeation experiment and therefore it is not possible to calculate parameters that are usually true coefficients or definite values. In this case, non-modelistic parameters can be used to estimate an enhancing effect.

In recent years, great efforts have been devoted to the design of drug delivery systems. Many polymeric excipients have been studied in order to make drug release fit the desired profiles. The aim of this work was to design a morphine oral suspension, as sustained release pharmaceutical formulations. To this end, two different ethylcellulose suspensions were prepared: one with the drug incorporated during synthesis (suspension A) so that the drug was inside the polymeric microparticles. In the second group of suspensions the drug was incorporated after synthesis (suspension B), thus resulting in the drug being adsorbed on the surface. The analytical technique used, spectrophotometry, showed that suspensions A were able to spontaneously encapsulate approximately 92% of the drug, whereas suspensions B adsorbed only 15% dose on the particle surface. Moreover, the diffusion results obtained with Franz-cells showed that suspensions A offered the possibility of easy control of the release rate of the active substance. This system transfers morphine hydrocloride during 24 h in accordance with a Weibull kinetic model. This dosage form presents the clinical advantage of less frequent dosing, with increased quality of life for patients. This report documents the suitability of our ethylcellulose polymeric suspension for encapsulated morphine with a controlled release rate.

The aim of this study was to improve the transdermal permeation of sodium diclofenac. Permeation studies were carried out in vitro using human skin (0.4 mm thick) from plastic surgery as a membrane. Four liquid formulations of 1% (w/w) sodium diclofenac were assayed: three ternary solvent systems (M4, M5, M6) and one microemulsion (M3). A 1% (w/w) solution of sodium diclofenac and a commercially available semisolid preparation were tested as reference formulations. The following permeation parameters for diclofenac were assessed: permeability coefficient, flux and drug permeated and retained in the skin at 24 h. The highest values of these parameters were obtained with formula M4, which contains transcutol® 59.2%, oleic acid 14.9% and d-limonene 5% (w/w) as permeation enhancers. The anti-inflammatory activity of this formula was compared with that of the semisolid preparation on carrageenan-induced paw edema in rats. As expected from in vitro results, the M4 diclofenac delivery system showed higher activity than the semisolid preparation, both when applied locally (to the inflammation area) and when applied systemically (to the back). Neither treatment irritated the skin when tested on rabbits in a 72-h trial. These results suggest that topical delivery of sodium diclofenac with an absorption enhancer such as a mixture of oleic acid and d-limonene (M4) may be an effective medication for both dermal and subdermal injuries.

Metallothioneins (MTs) are low molecular weight, cysteine-rich proteins, with an exceptional heavy metal coordination capacity. These metal chelating peptides play an active role in zinc homeostasis. Although their high heterogeneity, structural and functional studies have been mainly devoted to vertebrate and fungal MTs. Participation of metal ligands other than Cys and the presence of secondary structure elements in metal-MT complexes are fairly unknown, especially in non-vertebrate MTs. Six in vivo-synthesized Zn-MTs, representative of different MT families (mollusc, insect, nematode, echinoderm, plant and vertebrate) were studied by analytic and spectroscopic techniques. The examined MTs contain from 43 to 73 amino acids, among which at least a 30% are Cys, whereas very few are aromatic residues. Zn(II)-MT complexes were heterologously synthesized in E.coli, to obtain aggregates representative of those formed in biological systems. These complexes contain variable amounts of metal and sulfide ions, quantitatively evaluated by analytical measurements. The formation of more than one species (S2-containing and S2-devoid complexes), revealed by ESI-MS spectra, is a constant for all these MTs. The analysis of the Raman spectra gave information about the secondary structure of the metal-MT aggregates. For all the MT examined, a relevant contribution of beta-turns was shown, whereas in all cases the alfa-helix content resulted almost negligible. As regards Cys sulfurs, almost all Cys residues present in MTs resulted to be involved in the metal coordination, as indicated by the presence of several bands attributable to the metal-S stretching modes at low wavenumbers (< 500 cm-1). The high number of vM-S bands and their broadening suggest the formation of different metal centres. Significantly, Raman bands useful as markers of sulfide bridging ligands were identified. In Zn-CeMT2 (from CElegans) and Zn-QsMT (from plant - Quercus suber) the eventual participation of the His residue in metal binding was evaluated through a curve fitting analysis of the 1630-1565 cm-1 Raman spectral range. His residues resulted to be mainly coordinated in Zn-CeMT2 (~ 90%), whereas in Zn-QsMT His is mainly present as free tautomer (~ 90%). Raman spectroscopy showed to be a powerful technique, able to provide information on the state of the Cys sulfur (metal coordinated and oxidised), the metal binding environment (i.e. the participation of His in metal coordination), as well as on the secondary structure.

BACKGROUND AND AIMS: The TERMINAL FLOWER 1 (TFL1) gene is pivotal in the control of inflorescence architecture in arabidopsis. Thus, tfl1 mutants flower early and have a very short inflorescence phase, while TFL1-overexpressing plants have extended vegetative and inflorescence phases, producing many coflorescences. TFL1 is expressed in the shoot meristems, never in the flowers. In the inflorescence apex, TFL1 keeps the floral genes LEAFY (LFY) and APETALA1 (AP1) restricted to the flower, while LFY and AP1 restrict TFL1 to the inflorescence meristem. In spite of the central role of TFL1 in inflorescence architecture, regulation of its expression is poorly understood. This study aims to expand the understanding of inflorescence development by identifying and studying novel TFL1 regulators. METHODS: Mutagenesis of an Arabidopsis thaliana line carrying a TFL1::GUS (beta-glucuronidase) reporter construct was used to isolate a mutant with altered TFL1 expression. The mutated gene was identified by positional cloning. Expression of TFL1 and TFL1::GUS was analysed by real-time PCR and histochemical GUS detection. Double-mutant analysis was used to assess the contribution of TFL1 to the inflorescence mutant phenotype. KEY RESULTS: A mutant with both an increased number of coflorescences and high and ectopic TFL1 expression was isolated. Cloning of the mutated gene showed that both phenotypes were caused by a mutation in the ARGONAUTE1 (AGO1) gene, which encodes a key component of the RNA silencing machinery. Analysis of another ago1 allele indicated that the proliferation of coflorescences and ectopic TFL1 expression phenotypes are not allele specific. The increased number of coflorescences is suppressed in ago1 tfl1 double mutants. CONCLUSIONS: The results identify AGO1 as a repressor of TFL1 expression. Moreover, they reveal a novel role for AGO1 in inflorescence development, controlling the production of coflorescences. AGO1 seems to play this role through regulating TFL1 expression.

Brief episodes of tachycardia without myocardial ischemia prior to a coronary occlusion decrease myocardial infarct size in dogs. This non-ischemic preconditioning is mediated by adenosine. Because ischemic preconditioning is mediated through ATP dependent potassium channels, particularly the mitochondrial ones, we studied whether non-ischemic preconditioning is also mediated through these channels. In anesthetized dogs heart rate was kept constant at 120 cycles/min and aortic pressure changes were damped. Myocardial infarction was induced by occlusion of the anterior descending coronary artery for 60 min and reperfusion for 270 min. In a control group the infarct size (necrotic volume/risk region volume × 100) was 15.8 ± 1.5%. Preconditioning with five periods of tachycardia, 5 min in duration each at 213 cycles/min with intervening periods of 5 min of basal heart rate at 120 cycles/min, reduced the infarct size by 45.6% (p < 0.05) with respect to the control group. This effect was completely reverted by the blockade of ATP dependent potassium channels with glibenclamide or 5 hydroxydecanoate (a specific blocker of mitochondrial ATP dependent potassium channels) prior to preconditioning. These effects were not due to differences in collateral flow, risk region size or hemodynamic variables between the groups. These results show that mitochondrial ATP dependent potassium channels mediate non-ischemic preconditioning by tachycardia in dogs.