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Background The adoption of oncoplastic surgery in North America is poor despite evidence supporting the benefits. Surgeons take courses to acquire oncoplastic techniques, however, the effect of these courses is unknown. This study aimed to assess the impact of a hands-on oncoplastic course on surgeons’ comfort with oncoplastic techniques and rate of adoption of these techniques in their practice. Material and methods An online 10-question survey was developed and distributed to surgeons who had participated in a hands-on oncoplastic course offered in Ontario, Canada. Categorical data were reported using frequencies and percentages. Results A total of 105 surveys were sent out of which 69 attending surgeons responded (response rate: 65.7%). All respondents stated cosmesis was of the utmost importance in breast conserving surgery. The most common oncoplastic techniques they currently use included glandular re-approximation (98.4%), undermining of skin (93.6%), undermining of the nipple areolar complex (63.4%), and de-epithelialization and repositioning of the nipple areola complex (49.2%). Only 26% of respondnets stated they used more advanced techniques such as mammoplasty. Sixty percent of surgeons reported they used oncoplastic techniques in at least half of their cases. Ninety-two percent of respondents stated that the hands-on course increased the amount of oncoplastic techniques in their practice. At least 70% of respondents stated they would do another hands-on course. The main factor that facilitated the uptake of oncoplastic techniques was a better understanding of surgical techniques and planning. Conclusion A hands-on oncoplastic course helps surgeons adopt oncoplastic surgery techniques into their clinical practice. This teaching model allows surgeons to become comfortable with a variety of techniques. This study supports the relevance of a hands-on oncoplastic course to enhance the availability of safe oncoplastic surgery for breast cancer patients.

International audience; SIGNIFICANCE: Previous studies found no correlation between visual acuity and optical quality, in a population of young subjects with good vision. Using sinusoidal gratings, we systematically investigate the correlation between contrast sensitivity and optical quality as a function of spatial frequency. PURPOSE: This study describes the correlation between the contrast sensitivity function (CSF) and the modulation transfer function (MTF) in a sample of young and informed subjects. Our results are compared to prior studies on the correlation between visual acuity and metrics of image quality. We also compare our results to previous studies that compare the CSF, the MTF, and the neural contrast sensitivity function (NCSF). METHODS: The CSF of 28 informed subjects is measured in photopic conditions. The polychromatic MTF is computed from the measurements of monochromatic aberrations. The (CSF, MTF) correlation is estimated as the Pearson correlation coefficient, at each spatial frequency. The NCSF of each subject is estimated as the ratio of CSF to MTF. RESULTS: We obtain high correlation coefficients (0.8) in the 3-6 cycles per degree range of spatial frequencies, which also corresponds to high NCSF. Correlation decreases with increasing spatial frequency in the 6-18 cycles per degree range (down to 0.0 at 18 cycles per degree). In that range, optical and neural contrast sensitivities are both approximately reduced by factor 4. CONCLUSIONS: In our sample of young subjects with good vision, the contrast sensitivity function with sinusoidal gratings better differentiates eyes of good optical quality at intermediate spatial frequencies (3-6 cycles per degree) than at higher spatial frequencies (12-18 cycles per degree). At the highest tested spatial frequency of sinusoidal gratings (18 cycles per degree), there is no significant correlation between optical quality and contrast sensitivity.

Background: A phase 3 trial, ADVANCE (NCT03777059), demonstrated that atogepant, an oral, CGRP receptor antagonist dosed once daily, results in clinically meaningful reductions in mean monthly migraine days. This open-label extension for ADVANCE trial completers evaluated long-term safety and tolerability of atogepant over 40-weeks. Methods: Participants in this trial (NCT03939312), rolled over from the ADVANCE trial, were treated with atogepant 60mg once daily for 40-weeks, with a 4-week safety follow-up. Only safety data were collected. Results: 685 participants took at least one dose of study drug, 74.6% completed the 40-week treatment period; mean age of 41.8 years, 88.2% female, 84.4% white, and mean BMI of 30.58 kg/m2. Mean (SD) treatment duration was 233.6 (89.32) days. 62.5% of participants experienced a treatment-emergent adverse event (TEAE), with 8.8% considered treatment-related by the investigator; serious adverse events (SAEs) occurred in 3.4% of participants, none were treatment-related. The most frequent AEs leading to discontinuation was nausea (0.4%, n=3); the most frequent TEAEs observed included upper respiratory tract infection (5.5%, n=38) and urinary tract infection (5.3%, n=36). No deaths or hepatic safety issues were observed. Conclusions: Safety results are consistent with known safety profile of atogepant and support long-term safety and tolerability of once daily dosing of atogepant 60mg.

Background: In phase 3 ADVANCE, atogepant 60mg reduced mean monthly migraine days (MMDs) from 7.8 days (baseline) to 3.0 (weeks 9-12; Δ=−4.7) in the overall episodic migraine population [treatment responders and nonresponders (i.e., marked benefit and minimal benefit)], which obscures information regarding magnitude of treatment effect in these populations. Here, magnitude of treatment effect in atogepant responders and nonresponders is characterized. Methods: Mean MMDs, acute medication use-days (MUDs), and Migraine-Specific Quality of Life-Role Function-Restrictive (MSQ-RFR) scores were calculated in treatment responders (based on MMD percentage reduction) and nonresponders from ADVANCE participants. Results: From baseline to weeks 9-12, ≥50% improvement was achieved by 71% (139/195) of participants. In these responders, MMDs reduced from 7.6 to 1.3 (Δ=−6.3). 50% (97/195) of participants achieved ≥75% response. In this group, MMDs reduced from 7.7 to 0.6 (Δ=−7.1). Atogepant 60mg nonresponders (

Angiotensin Converting Enzyme 2 (ACE2) is the human receptor that interacts with the Spike protein of coronaviruses, including the one that produced the 2020 coronavirus pandemic (COVID-19). Thus, ACE2 is a potential target for drugs that disrupt the interaction of human cells with SARS-CoV-2 to abolish infection. There is also interest on drugs that inhibit or activate ACE2, i.e. for cardiovascular disorders or colitis. Compounds binding at alternative sites could allosterically affect the interaction with Spike protein. We here review biochemical, chemical biology and structural information on ACE2, including the recent cryoEM structures of full length ACE2. We conclude that ACE2 is very dynamic and that allosteric drugs may be developed to target ACE2. At the time of the 2020 pandemic, we suggest that available ACE2 inhibitors or activators in advanced development should be tested for their ability to allosterically displace the interaction between ACE2 and the Spike protein. Fil: Gross, Lissy Zoe Florens. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigación en Biomedicina de Buenos Aires - Instituto Partner de la Sociedad Max Planck; Argentina Fil: Sacerdoti, Mariana. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigación en Biomedicina de Buenos Aires - Instituto Partner de la Sociedad Max Planck; Argentina Fil: Piiper, Albrecht. Goethe Universitat Frankfurt; Alemania Fil: Zeuzem, Stefan. Goethe Universitat Frankfurt; Alemania Fil: Leroux, Alejandro Ezequiel. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigación en Biomedicina de Buenos Aires - Instituto Partner de la Sociedad Max Planck; Argentina Fil: Biondi, Ricardo Miguel. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigación en Biomedicina de Buenos Aires - Instituto Partner de la Sociedad Max Planck; Argentina

The group of AGC kinases consists of 63 evolutionarily related serine/threonine protein kinases comprising PDK1, PKB/Akt, SGK, PKC, PRK/PKN, MSK, RSK, S6K, PKA, PKG, DMPK, MRCK, ROCK, NDR, LATS, CRIK, MAST, GRK, Sgk494, and YANK, while two other families, Aurora and PLK, are the most closely related to the group. Eight of these families are physiologically activated downstream of growth factor signalling, while other AGC kinases are downstream effectors of a wide range of signals. The different AGC kinase families share aspects of their mechanisms of inhibition and activation. In the present review, we update the knowledge of the mechanisms of regulation of different AGC kinases. The conformation of the catalytic domain of many AGC kinases is regulated allosterically through the modulation of the conformation of a regulatory site on the small lobe of the kinase domain, the PIF-pocket. The PIF-pocket acts like an ON-OFF switch in AGC kinases with different modes of regulation, i.e. PDK1, PKB/Akt, LATS and Aurora kinases. In this review, we make emphasis on how the knowledge of the molecular mechanisms of regulation can guide the discovery and development of small allosteric modulators. Molecular probes stabilizing the PIF-pocket in the active conformation are activators, while compounds stabilizing the disrupted site are allosteric inhibitors. One challenge for the rational development of allosteric modulators is the lack of complete structural information of the inhibited forms of full-length AGC kinases. On the other hand, we suggest that the available information derived from molecular biology and biochemical studies can already guide screening strategies for the identification of innovative mode of action molecular probes and the development of selective allosteric drugs for the treatment of human diseases. Fil: Leroux, Alejandro Ezequiel. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigación en Biomedicina de Buenos Aires - Instituto Partner de la Sociedad Max Planck; Argentina Fil: Schulze, Jörg. Universitätsklinikum Frankfurt; Alemania Fil: Biondi, Ricardo Miguel. Universitätsklinikum Frankfurt; Alemania. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigación en Biomedicina de Buenos Aires - Instituto Partner de la Sociedad Max Planck; Argentina. German Cancer Consortium; Alemania

Infectious diseases are unpredictable, with heterogeneous clinical presentations, diverse pathogens, and various susceptibility rates to anti-infective agents. These features lead to a wide variety of clinical practices, which in turn strongly limits their evaluation. We have been using a medical table since 2005 to monitor the medical activity in our department. The observation of heterogeneous therapeutic practices led to drafting up our own antibiotic guidelines and to implementing a continuous evaluation of their observance and impact on morbidity and mortality associated with infectious diseases, including adverse effects of antibiotics, duration of hospital stay, use of intensive care, and deaths. The 10-year analysis of medical practices using the medical table is based on more than 10,000 hospitalizations. It shows simplified antibiotic therapies and a reduction in infection-related morbidity and mortality. The medical table is a major tool for antimicrobial stewardship, leading to constant benefits for patients.

Allostery is a basic principle that enables proteins to process and transmit cellular information. Protein kinases evolved allosteric mechanisms to transduce cellular signals to downstream signalling components or effector molecules. Protein kinases catalyse the transfer of the terminal phosphate from ATP to protein substrates upon specific stimuli. Protein kinases are targets for the development of small molecule inhibitors for the treatment of human diseases. Drug development has focussed on ATP-binding site, while there is increase interest in the development of drugs targeting alternative sites, i.e. allosteric sites. Here, we review the mechanism of regulation of protein kinases, which often involve the allosteric modulation of the ATP-binding site, enhancing or inhibiting activity. We exemplify the molecular mechanism of allostery in protein kinases downstream of PI3-kinase signalling with a focus on phosphoinositide-dependent protein kinase 1 (PDK1), a model kinase where small compounds can allosterically modulate the conformation of the kinase bidirectionally.

Background There has been a rise in obstetric intervention during labour and birth including rates of caesarean section around the world. The rates of caesarean section are lower in France (20%) than Australia (34%). Both countries have universal health care systems. Australia has a well-established program of obstetric benchmarking and national data collection for comparing maternal and perinatal outcomes taking into account maternity units and maternal characteristics. Although the optimal caesarean section rate is not known, variation in caesarean section rates raise questions about what is driving variation in practice and whether the right care is being delivered. The World Health Organisation has recently released non-clinical recommendations aiming to reduce unnecessary caesarean sections, including that births are classified by Robson classification for easier comparison. Methods We aim to compare rates of mode of birth in 2 tertiary hospitals- one in France and one in Australia using Robson classification. This project will investigate differences in patient casemix, and obstetric management that may lead to differences in outcome. In addition, we will try to explore differences in the health systems including clinical activity, induction rates, staffing (number, type (medical, nursing, midwifery)), and models of care. Outcomes Caesarean section is one of the most common operations performed in hospitals. Important variations of surgical rates raise question regarding whether access to this procedure is optimized. Determining current practice and outcomes of women in two different settings will provide important data to inform hospital practice and further research. In addition, this research may inform national guidelines about indications for caesarean section.

Protein–protein interactions often regulate the activity of protein kinases by allosterically modulating the conformation of the ATP-binding site. Bidirectional allostery implies that reverse modulation (i.e., from the ATP-binding site to the interaction and regulatory sites) must also be possible. Here, we review both the allosteric regulation of protein kinases and recent work describing how compounds binding at the ATP-binding site can promote or inhibit protein kinase interactions at regulatory sites via the reverse mechanism. Notably, the pharmaceutical industry has been developing compounds that bind to the ATP-binding site of protein kinases and potently disrupt protein–protein interactions between target protein kinases and their regulatory interacting partners. Learning to modulate allosteric processes will facilitate the development of protein–protein interaction modulators.

Leishmania parasites cause a broad spectrum of clinical manifestations in humans and the available clinical treatments are far from satisfactory. Since these pathogens require large amounts of NADPH to maintain intracellular redox homeostasis, oxidoreductases that catalyze the production of NADPH are considered as potential drug targets against these diseases. In the sequenced genomes of most Leishmania spp. two putative malic enzymes (MEs) with an identity of about 55% have been identified. In this work, the ME from L. major (LmjF24.0770, Lmj_ME-70) and its less similar homolog from L. mexicana (LmxM.24.0761, Lmex_ME-61) were cloned and functionally characterized. Both MEs specifically catalyzed NADPH production, but only Lmex_ME-61 was activated by L-aspartate. Unlike the allosterically activated human ME, Lmex_ME-61 exhibited typical hyperbolic curves without any sign of cooperativity in the absence of L-aspartate. Moreover, Lmex_ME-61 and Lmj_ME-70 differ from higher eukaryotic homologs in that they display dimeric instead of tetrameric molecular organization. Homology modeling analysis showed that Lmex_ME-61 and Lmj_ME-70 notably differ in their surface charge distribution; this feature encompasses the coenzyme binding pockets as well. However, in both isozymes, the residues directly involved in the coenzyme binding exhibited a good degree of conservation. Besides, only Lmex_ME-61 and its closest homologs were immunodetected in cell-free extracts from L. mexicana, L. amazonensis and L. braziliensis promastigotes. Our findings provide a first glimpse into the biochemical properties of leishmanial MEs and suggest that MEs could be potentially related to the metabolic differences among the species of Leishmania parasites. Fil: Giordana, Lucila. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Química y Físico-Química Biológicas ; Argentina Fil: Sosa, Máximo Hernán. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Farmacológicas. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Investigaciones Farmacológicas; Argentina Fil: Leroux, Alejandro Ezequiel. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigación en Biomedicina de Buenos Aires - Instituto Partner de la Sociedad Max Planck; Argentina Fil: Mendoza Rodas, Elkin F.. Universidade Estadual Paulista Julio de Mesquita Filho; Brasil Fil: Petray, Patricia Beatriz. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones en Microbiología y Parasitología Médica. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones en Microbiología y Parasitología Médica; Argentina Fil: Nowicki, Cristina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Química y Físico-Química Biológicas ; Argentina

Most known thioredoxin-type proteins (Trx) participate in redox pathways, using two highly conserved cysteine residues to catalyze thiol-disulfide exchange reactions. Here we demonstrate that the so far unexplored Trx2 from African trypanosomes (Trypanosoma brucei) lacks protein disulfide reductase activity but functions as an effective temperature-activated and redox-regulated chaperone. Immunofluorescence microscopy and fractionated cell lysis revealed that Trx2 is located in the mitochondrion of the parasite. RNA-interference and gene knock-out approaches showed that depletion of Trx2 impairs growth of both mammalian bloodstream and insect stage procyclic parasites. Procyclic cells lacking Trx2 stop proliferation under standard culture conditions at 27°C and are unable to survive prolonged exposure to 37°C, indicating that Trx2 plays a vital role that becomes augmented under heat stress. Moreover, we found that Trx2 contributes to the in vivo infectivity of T. brucei. Remarkably, a Trx2 version, in which all five cysteines were replaced by serine residues, complements for the wildtype protein in conditional knock-out cells and confers parasite infectivity in the mouse model. Characterization of the recombinant protein revealed that Trx2 can coordinate an iron sulfur cluster and is highly sensitive towards spontaneous oxidation. Moreover, we discovered that both wildtype and mutant Trx2 protect other proteins against thermal aggregation and preserve their ability to refold upon return to non-stress conditions. Activation of the chaperone function of Trx2 appears to be triggered by temperature-mediated structural changes and inhibited by oxidative disulfide bond formation. Our studies indicate that Trx2 acts as a novel chaperone in the unique single mitochondrion of T. brucei and reveal a new perspective regarding the physiological function of thioredoxin-type proteins in trypanosomes., Author summary African trypanosomes are the causative agents of human sleeping sickness and Nagana cattle disease. These strictly extracellular pathogens multiply in the blood and body fluids of their mammalian hosts and the tsetse fly vector, where efficient redox regulation is essential for parasite survival. While most organisms use the glutathione/glutathione reductase and thioredoxin/thioredoxin reductase couples to maintain cellular redox balance, trypanosomes rely on a unique trypanothione-based thiol metabolism to survive exogenous and endogenous oxidative stresses. Despite the lack of thioredoxin reductases, the Trypanosoma brucei genome encodes thioredoxins, raising questions for their biological function. Our work is the first report on T. brucei thioredoxin-2 (Trx2). We show that Trx2 is located in the mitochondrion and its absence affects parasite proliferation and infectivity. Recombinant Trx2 lacks protein disulfide reductase activity but protects proteins against aggregation and maintains them folding-competent. Remarkably, a mutant that is devoid of any cysteine residues is able to fully substitute for the authentic protein under in vitro and in vivo conditions. Our data reveal that Trx2 does not function as a classical thioredoxin but acts as a chaperone that plays a crucial role in the mitochondrion of T. brucei.

Introduction Chez l’enfant, l’implication de la toxine de leucocidine de Panton-Valentine (LPV + ) dans les infections invasives communautaires (IIC) a Staphylococcus aureus (S. aureus) est tres souvent suspectee et pose la question de l’antibiotherapie probabiliste en raison du risque accru de resistance a la meticilline ainsi que la question de l’ajout d’un antibiotique anti-toxinique. Cependant, l’epidemiologie de la resistance des S. aureus LPV + sur des souches invasives en France est ancienne et la virulence de la toxine debattue dans la litterature recente. Le laboratoire de microbiologie de notre hopital a, durant des annees, recherche la presence de la toxine de maniere quasi exhaustive sur les prelevements profonds. Le but de cette etude est de decrire les IIC a S. aureus LPV + chez l’enfant et de definir les facteurs de risque pediatriques predictifs de production de LPV. Materiels et methodes Nous avons conduit une etude cas-temoins entre 2012 et 2016 monocentrique pediatrique en incluant les IIC a S. aureus LPV + (definie par un prelevement profond isolant S. aureus LPV + ), et en appariant un temoin avec IIC a S. aureus LPV – pour un cas d’IIC a S. aureus LPV + sur l’âge, le sexe et le type d’infection. Resultats Nous avons identifie 78 cas d’IIC a S. aureusLPV+ parmi lesquelles 64,1 % d’infections cutanees et du tissu mou (n = 50), 21,8 % de bacteriemies (n = 17), 7,7 % d’osteo-arthrites (n = 6), 5,3 % de pneumonies (n = 4) et 1,3 % de pleuropneumonies (n = 1). Apres appariement, la presence de signes de gravite etait comparable dans les 2 groupes (9 %, p = 0,85), de meme que la presentation clinique initiale (p = 0,18) et la duree mediane d’hospitalisation (5 jours, p = 0,42). Un S. aureus resistant a la meticilline (SARM) etait present dans 35,9 % (n = 28/78) des IIC a S. aureus LPV + contre 12,7 % (n = 9/71) dans le groupe PLV – (p = 0,001). Les facteurs predictifs de production de LPV etaient une furonculose personnelle ou familiale (p = 0,01), un voyage a l’etranger (p = 0,03), des parents originaires d’Asie ou d’Afrique sub-saharienne et la presence de 3 sites infectieux ou plus (p = 0,02). La presence d’un SARM (p = 0,001) et le profil kanamycine-tetracycline-acide fusidique resistant (p = 0,004) etaient aussi associes a la LPV. Conclusion Les IIC a S. aureus LPV + pediatriques sont donc majoritairement des infections cutanees et du tissu mou et la production de LPV n’est pas un facteur de gravite d’apres notre etude. L’association LPV et SARM est a prendre en compte pour le choix du traitement antibiotique probabiliste des IIC a S. aureus LPV + chez l’enfant.

Background: Trypanothione synthetase catalyzes the conjugation of spermidine with two GSH molecules to form trypanothione. Results: The kinetic parameters were measured under in vivo-like conditions. A mathematical model was developed describing the entire kinetic profile. Conclusion: Trypanothione synthetase is affected by substrate and product inhibition. Significance: The combined kinetic and modeling approaches provided a so far unprecedented insight in the mechanism of this parasite-specific enzyme., In pathogenic trypanosomes, trypanothione synthetase (TryS) catalyzes the synthesis of both glutathionylspermidine (Gsp) and trypanothione (bis(glutathionyl)spermidine (T(SH)2)). Here we present a thorough kinetic analysis of Trypanosoma brucei TryS in a newly developed phosphate buffer system at pH 7.0 and 37 °C, mimicking the physiological environment of the enzyme in the cytosol of bloodstream parasites. Under these conditions, TryS displays Km values for GSH, ATP, spermidine, and Gsp of 34, 18, 687, and 32 μm, respectively, as well as Ki values for GSH and T(SH)2 of 1 mm and 360 μm, respectively. As Gsp hydrolysis has a Km value of 5.6 mm, the in vivo amidase activity is probably negligible. To obtain deeper insight in the molecular mechanism of TryS, we have formulated alternative kinetic models, with elementary reaction steps represented by linear kinetic equations. The model parameters were fitted to the extensive matrix of steady-state data obtained for different substrate/product combinations under the in vivo-like conditions. The best model describes the full kinetic profile and is able to predict time course data that were not used for fitting. This system's biology approach to enzyme kinetics led us to conclude that (i) TryS follows a ter-reactant mechanism, (ii) the intermediate Gsp dissociates from the enzyme between the two catalytic steps, and (iii) T(SH)2 inhibits the enzyme by remaining bound at its product site and, as does the inhibitory GSH, by binding to the activated enzyme complex. The newly detected concerted substrate and product inhibition suggests that TryS activity is tightly regulated.

In mice cynaropicrin (CYN) potently inhibits the proliferation of Trypanosoma brucei-the causative agent of Human African Trypanosomiasis-by a so far unknown mechanism. We hypothesized that CYNs α,β-unsaturated methylene moieties act as Michael acceptors for glutathione (GSH) and trypanothione (T(SH)2), the main low molecular mass thiols essential for unique redox metabolism of these parasites. The analysis of this putative mechanism and the effects of CYN on enzymes of the T(SH)2 redox metabolism including trypanothione reductase, trypanothione synthetase, glutathione-S-transferase, and ornithine decarboxylase are shown. A two step extraction protocol with subsequent UPLC-MS/MS analysis was established to quantify intra-cellular CYN, T(SH)2, GSH, as well as GS-CYN and T(S-CYN)2 adducts in intact T. b. rhodesiense cells. Within minutes of exposure to CYN, the cellular GSH and T(SH)2 pools were entirely depleted, and the parasites entered an apoptotic stage and died. CYN also showed inhibition of the ornithine decarboxylase similar to the positive control eflornithine. Significant interactions with the other enzymes involved in the T(SH)2 redox metabolism were not observed. Alongside many other biological activities sesquiterpene lactones including CYN have shown antitrypanosomal effects, which have been postulated to be linked to formation of Michael adducts with cellular nucleophiles. Here the interaction of CYN with biological thiols in a cellular system in general, and with trypanosomal T(SH)2 redox metabolism in particular, thus offering a molecular explanation for the antitrypanosomal activity is demonstrated. At the same time, the study provides a novel extraction and analysis protocol for components of the trypanosomal thiol metabolism.

Parasitic infections continue to be a major problem for global human health. Vaccines are practically not available and chemotherapy is highly unsatisfactory. One approach toward a novel antiparasitic drug development is to unravel pathways that may be suited as future targets. Parasitic organisms show a remarkable diversity with respect to the nature and functions of their main low-molecular-mass antioxidants and many of them developed pathways that do not have a counterpart in their mammalian hosts.Work of the last years disclosed the individual antioxidants employed by parasites and their distinct pathways. Entamoeba, Trichomonas, and Giardia directly use cysteine as main low-molecular-mass thiol but have divergent cysteine metabolisms. Malarial parasites rely exclusively on cysteine uptake and generate glutathione (GSH) as main free thiol as do metazoan parasites. Trypanosomes and Leishmania have a unique trypanothione-based thiol metabolism but employ individual mechanisms for their cysteine supply. In addition, some trypanosomatids synthesize ovothiol A and/or ascorbate. Various essential parasite enzymes such as trypanothione synthetase and trypanothione reductase in Trypanosomatids and the Schistosoma thioredoxin GSH reductase are currently intensively explored as drug target molecules.Essentiality is a prerequisite but not a sufficient property of an enzyme to become a suited drug target. The availability of an appropriate in vivo screening system and many other factors are equally important.The current organism-wide RNA-interference and proteome analyses are supposed to reveal many more interesting candidates for future drug development approaches directed against the parasite antioxidant defense systems.

For increasing the shelf life of strawberries during storage, bioactive coatings were applied using modified polysaccharides of chitosan. First, antimicrobial tests were performed with selected essential oils to evaluate their antimicrobial capacities against moulds and total flora isolated from strawberries. Red thyme (RT) and oregano extract (OR) were found as strong bioactive agents against moulds and total flora isolated from strawberries, whereas limonene (LIM) and peppermint (PM) had lower antimicrobial properties. These essential oils were also used as bioactive compounds which were sprayed onto strawberries and evaluated for their potential to increase shelf life during storage at 4 °C. RT, PM and LIM were found to be more efficient preservative agents for strawberries during 14 days of storage. Finally, chitosan was functionalized by acylation with palmitoyl chloride to increase its hydrophobicity, to ensure a controlled release and improve its stability and adhesion to the fruit product. LIM and PM were incorporated into the modified chitosan to create bioactive edible coatings and these were tested for their ability to extend the shelf life of fresh strawberries during storage. Formulations based on modified chitosan containing LIM and Tween®80 were shown to perform better than other formulations.

Fungus balls are a non-invasive form of fungal infection involving the maxillary sinus in most cases. Sphenoid sinus fungus balls (SSFB) are rare and their clinical presentation is not well described. We intended to define the clinical presentation of sphenoid fungus balls, and retrospectively reviewed 24 cases of SSFB seen at our institution over a 10-year period, identified through pathological reports. Presenting symptoms were separated into three groups: headache, rhinological and asymptomatic. Headaches were subdivided into acute and chronic, unilateral and diffuse. Radiological clues leading to diagnosis were reviewed. Prognosis was determined from medical files or by phone calls. Sixty-seven per cent of patients were female. The mean age at presentation was 65 years. Sixty-two per cent presented with headache, 36% unilateral, mainly in the first trigeminal branch territory. Rhinological symptoms were seen in 21%. In 16% of patients the SSFB was asymptomatic and found during routine tests. SSFB, even if noninvasive, did lead to recurrent bacterial infections and central nervous system complications in three patients. Of 15 patients presenting with headache, 10 were significantly improved post surgery. The prognosis is good, with no recurrence of fungal infection after a main follow-up of 2.3 years. Our study underlines that SSFB present with headaches, often unilateral and in the fronto-orbital region. Proper imaging of the sphenoid sinus is useful in patients with unexplained headache. The neurologist has to be aware of radiological clues suggesting fungal sinus infection, since surgery is the main treatment, with good prognosis and frequent resolution of headaches.

Trypanosomatids are the causative agents of African sleeping sickness, Chagas' disease, and the different forms of leishmaniasis. This family of protozoan parasite possesses a trypanothione-based redox metabolism that provides the reducing equivalents for various vital processes such as the biosynthesis of DNA precursors and the detoxification of hydroperoxides. Almost all enzymes of the redox pathway proved to be essential and therefore fulfil one crucial prerequisite for a putative drug target. Trypanothione synthetase and trypanothione reductase are present in all trypanosomatids but absent from the mammalian host which, in addition to the essentiality, renders them highly specific. The chemotherapy research on both enzymes is further supported by the availability of high-throughput screening techniques and crystal structures. In this review we focus on the recent advances and limitations in the development of lead compounds targeting trypanothione synthetase and trypanothione reductase. We present an overview of the available inhibitors and discuss future perspectives including other components of the parasite-specific redox pathway.

Placement analysis in complex antenna scenarios require accurate computational electromagnetic (CEM) tools. A fundamental requirement to achieve truthful results, is that the source antenna must be accurately modelled. However, in many practical cases, a full-wave representation of the physical antenna is unfeasible or unavailable in the format required by the desired CEM solver. This paper describes a procedure to derive, a computational efficient, full wave representation of an existing antenna for CEM solvers. The procedure is based on post-processing of the measured antenna pattern. The desired source antenna is measured while situated in a suitable environment, of reduced complexity and size, that locally resemble the final antenna environment. The measured field is expanded using equivalent currents and an equivalent near field (NF) source is derived in a volume conformal to the antenna. The NF source is a truthful representation of the measured source and highly suitable for CEM analysis. The procedure of generating the NF source from measurements is fully general and can be used for the installation of antennas in complex environments of arbitrary complexity. The procedure is illustrated and validated by an example concerning the placement of a monocone antenna (SMC2200) on an electrically large rectangular ground plane. The monocone antenna is initially measured on a small circular ground plane that resemble the final operational environment in the vicinity of the antenna. The measurement has been performed in a MVG multi probe spherical near field system. Simulations using the measured source on the larger rectangular structure have been compared with a full numerical model of the antenna and structure for verification. A further validation of the NF source method, based on the measurement of the larger rectangular structure is on-going.

Electromagnetic analysis of antennas in complex scenarios require accurate numerical modelling solvers. A fundamental requirement to achieve truthful results is the accurate representation/modelling of the source antenna. However, in many practical cases a full-wave representation of the physical antenna is either unfeasible or unavailable in the format required by the desired computational electromagnetic (CEM) tool. This paper describes an innovative procedure enabling measured antenna data to be used as numerical sources in commercially-available computational environments. The proposed method is based on the equivalent current representation of the antenna when measured in isolation or in a suitable environment locally representative of the full complex structure. The equivalent current representation is highly accurate, even when the antenna is placed in close proximity, or mounted directly on the complex structure. The method is fully general and can be used with general antenna in complex environments of arbitrary shape and complexity. The accuracy and effectiveness of the procedure has been illustrated by an example of an offset-fed reflector. The complete reflector system has been measured and compared with results from the proposed procedure using a measured feed and numerical modelling of the reflector. As further validation, the results have been compared to a dedicated computation analysis of the full antenna model.

The African trypanosome, Ttypanosoma brucei, is a unicellular parasite causing African Trypanosomiasis (sleeping sickness in humans and nagana in animals). Due to some of its unique properties, it has emerged as a popular model organism in systems biology. A predictive quantitative model of glycolysis in the bloodstream form of the parasite has been constructed and updated several times. The Silicon Trypanosome is a project that brings together modellers and experimentalists to improve and extend this core model with new pathways and additional levels of regulation. These new extensions and analyses use computational methods that explicitly take different levels of uncertainty into account. During this project, numerous tools and techniques have been developed for this purpose, which can now be used for a wide range of different studies in systems biology.

Comparative studies showed that, like Trypanosoma cruzi, Trypanosoma brucei exhibits functional cytosolic and mitochondrial malic enzymes (MEs), which are specifically linked to NADP. Kinetic studies provided evidence that T. cruzi and T. brucei MEs display similarly high affinities towards NADP(+) and are also almost equally efficient in catalyzing the production of NADPH. Nevertheless, in contrast to the cytosolic ME from T. cruzi, which is highly activated by l-aspartate (over 10-fold), the T. brucei homologue is slightly more active (50%) in the presence of this amino acid. In T. brucei, both isozymes appear to be clearly more abundant in the insect stage, although they can be immunodetected in the bloodstream forms. By contrast, in T. cruzi the expression of the mitochondrial ME seems to be clearly upregulated in amastigotes, whereas the cytosolic isoform appears to be more abundant in the insect stages of the parasite. It might be hypothesized that in those environments where glucose is very low or absent, these pathogens depend on NADP-linked dehydrogenases such as the MEs for NADPH production, as in those conditions the pentose phosphate pathway cannot serve as a source of essential reducing power.

Trypanosoma cruzi exhibits two putative isocitrate dehydrogenases (IDHs). Both idh genes were cloned and the recombinant enzymes expressed in Escherichia coli. Our results showed that T. cruzi IDHs are strictly dependent on NADP(+) and display apparent affinities towards isocitrate and the coenzyme in the low micromolar range. In T. cruzi, IDHs are cytosolic and mitochondrial enzymes, and there is no evidence for the typical Krebs cycle-related NAD-dependent IDH. Hence, like in Trypanosoma brucei, the Krebs cycle is not a canonical route in T. cruzi. However, the citrate produced in the mitochondrion could be isomerized into isocitrate in the cytosol and the mitochondrion by means of the putative aconitase, which would provide the substrate for both IDHs. The cytosolic IDH is significantly more abundant in amastigotes, cell-derived and metacyclic trypomastigotes than in epimastigotes. This observation fits in well with the expected oxidative burst this pathogen has to face when infecting the mammalian host.

A FM-UWB transceiver prototype operating in the 7.2 – 7.7 GHz band based upon 3 full-custom ICs is presented. Receiver sensitivity is −85 dBm for a BER of 10−6 at 62.5 kbps. A range of 15 meters is demonstrated under LOS conditions. Power consumption of a SoC version is estimated at 5 mW for the transmitter and 14 mW for the receiver. Subcarrier FDMA multiple access and robustness to narrowband interference are also demonstrated.

This paper introduces a procedure for multi-objective optimization in antenna design. The problem described consists of the optimization of a seven element antenna array to improve the radiating performance. The three objectives considered are the minimization of the return loss, together with the minimization of the cross polarization and the mutual coupling between the ports; simultaneously, the design should satisfy a complex constraint on the far-field pattern. Since there is no single optimum to be found, the MOGT and MOGA-II (Game Theory-based and Genetic Algorithm) were used as multi-objective algorithms. The optimization of the antenna was carried out by employing a parametric CST MICROWAVE STUDIOreg (CST MWS) model, performing a time domain analysis, and using ESTECOpsilas modeFRONTIER4reg multi-objective optimization and process integration tool. The distributed optimization search exploited the parallelization capabilities of the MOGT and MOGA-II algorithms, which allowed the simultaneous evaluation of several design configurations by running concurrent threads of the solver. The results obtained are very satisfactory, and the procedure described can be applied to even more complex antenna design problems.

The WiseNET system includes an ultra low-power system-on-chip (SoC) hardware platform and WiseMAC, a low power medium access control protocol (MAC) dedicated to duty-cycled radios. Both elements have been designed to meet the specific requirements of wireless sensor networks and are particularly well suited to ad-hoc and hybrid networks. The WiseNET radio offers dual-band operation (434-MHz and 868-MHz) and runs from a single 1.5-V battery. It consumes only 2.5-mW in receive mode with a sensitivity smaller than -108-dBm at a BER of 10-3 and for a 100-kb/s data rate. In addition to this low-power radio, the WiseNET system-on-chip (SoC) also includes all the functions required for data acquisition, processing and storage of the information provided by the sensors. Ultra-low power consumption with the WiseNET system is achieved thanks to the combination of the low power consumption of the transceiver and the high energy efficiency of the WiseMAC protocol. The WiseNET solution consumes more than 250 times less power than comparable solutions based on the IEEE 802.15.4 standard