The yeast ubiquitin ligase SCF(Met30): connecting environmental and intracellular conditions to cell division

Ubiquitination regulates a host of cellular processes and is well known for its role in progression through the cell division cycle. In budding yeast, cadmium and arsenic stress, the availability of sulfur containing amino acids, and the intracellular concentration of S-adenosylmethionine are linked to cell cycle regulation through the ubiquitin ligase SCF(Met30). Regulation is achieved by ubiquitination of the transcription factor Met4. Met4 activity is controlled by a regulatory K48-linked ubiquitin chain that is synthesized by Cdc34/SCF(Met30). A ubiquitin-interacting-motif (UIM) present in Met4 prevents degradation of ubiquitinated Met4 allowing the ubiquitin chain to function as a reversible switch of Met4 activity. Here we discuss mechanisms of Met4 and SCF(Met30 )regulation in response to intracellular and environmental conditions, and describe the integration of these signals with cell cycle control

HIV-1 diversity in viral reservoirs obtained from circulating T-cell subsets during early ART and beyond

Even during extended periods of effective immunological control, a substantial dynamic of the viral genome can be observed in different cellular compartments in HIV-1 positive individuals, indicating the persistence of active viral reservoirs. To obtain further insights, we studied changes in the proviral as well as in the viral HIV-1 envelope (Env) sequence along with transcriptional, translational and viral outgrowth activity as indicators for viral dynamics and genomic intactness. Our study identified distinct reservoir patterns that either represented highly sequence-diverse HIV-1 populations or only a single / few persisting virus variants. The single dominating variants were more often found in individuals starting ART during early infection phases, indicating that early treatment might limit reservoir diversification. At the same time, more sequence-diverse HIV reservoirs correlated with a poorer immune status, indicated by lower CD4 count, a higher number of regimen changes and more co-morbidities. Furthermore, we noted that in T-cell populations in the peripheral blood, replication-competent HIV-1 is predominantly present in Lymph node homing TN (naïve) and TCM (central memory) T cells. Provirus genomes archived in TTM (transitional memory) and TEM (effector memory) T cells more frequently tended to carry inactivating mutations and, population-wise, possess changes in the genetic diversity. These discriminating properties of the viral reservoir in T-cell subsets may have important implications for new early therapy strategies, underscoring the critical role of early therapy in preserving robust immune surveillance and constraining the viral reservoir

Validação e aplicabilidade do software Wounds Monitoring na avaliação e monitoramento de feridas

The aim was to validate an application software for recording, monitoring and evaluating wounds. Methodological, applied, quantitative study. The references adopted in this project were described in Standard No. 25010 of the International Organization for Standardization/International Electrotechnical Commission. The results were analyzed by a rule proposed by the Brazilian Association of Technical Norms in its Normative No. 14598-6. The feasibility of the system was evaluated through a test of the prototype, with the group of evaluators that was composed of two categories: specialists in information technology and nurses. In the evaluation of the nurses' expertise, the characteristic functional adequacy, reliability, usability, efficiency in performance, compatibility, and safety, obtained 100% of the responses in agreement. Professional IT experts also rated maintainability with 97.3% reliability with 84.9%, usability with 84.6% of the answers in agreement. The software was validated, allowing evaluating the technical quality and functional performance applied to the evaluation and monitoring of wounds.Objetivou-se validar um software aplicativo para registro, monitoramento e avaliação de feridas. Estudo metodológico, aplicado, quantitativo. As referências adotadas neste projeto foram descritas na Norma n.º 25010 da Internacional Organization for Standardization/International Electrotechnical Commission. Os resultados foram analisados por regra proposta pela Associação Brasileira de Normas Técnicas em sua Normativa n.º 14598-6. Foi avaliada a viabilidade do sistema através de um teste do protótipo, com o grupo de avaliadores que foi composto por duas categorias: especialistas em tecnologia da informação e enfermeiros. Na avaliação dos expertises enfermeiros, a característica adequação funcional confiabilidade, usabilidade, eficiência no desempenho, compatibilidade, e segurança, obtiveram 100% das respostas em acordo. Os expertises profissionais de informática também avaliaram manutenibilidade com 97,3% confiabilidade com 84,9%, usabilidade com 84,6% das respostas de acordo. O software foi validado permitindo avaliar a qualidade técnica e desempenho funcional aplicado à avaliação e monitoramento de feridas

Contribuição de um software para o registro, monitoramento e avaliação de feridas

The aim was to describe the construction of a software for recording, monitoring, and evaluating wounds. It is a technological production research containing software development. It was carried out in three stages: project management through the Scrum tool, software construction, content validation and pre-test. As a result, the app was built after reviewing the literature in databases. The management system allowed registering patients, performing the assessment, and monitoring of the evolution of the lesions through images and other tools, such as graphs of the results of the Braden Scale for Predicting Pressure Ulcer Risk and Pressure Ulcer Scale for Healing. In addition, information about coverage and treatment alternatives is available. The development of an initial prototype allowed the exploration of ideas, before investing in production, helping to save time and resources, modulating the final product to be developed according to the needs of the target audience. In conclusion, the present study demonstrated that the construction of the software meets the need for recording, monitoring, and evaluating wounds. The prospects are that it can also be used to carry out realistic simulation and training, in addition to having easy access and handling.Objetivou-se descrever a construção de um software para registro, monitoramento e avaliação de feridas. Trata-se de uma pesquisa de produção tecnológica contendo o desenvolvimento de software. Foi concretizado em três etapas: gestão do projeto através da ferramenta Scrum, construção do software, validação de conteúdo e pré-teste. Como resultado aplicativo foi construído após revisão da literatura em bases de dados. O sistema de gestão permitiu cadastrar pacientes, realizar a avaliação e o monitoramento da evolução das lesões através de imagens e de outras ferramentas, tais como gráficos de resultados das escalas Braden Scale for Predicting Pressure Ulcer Risk e Pressure Ulcer Scale for Healing. Além disso, estão disponíveis informações sobre coberturas e alternativas para o tratamento. O desenvolvimento de um protótipo inicial permitiu a exploração das ideias, antes do investimento na produção, auxiliando na economia de tempo e recursos, modulando de acordo com as necessidades do público-alvo, o produto final a ser desenvolvido. Por conclusão, o presente estudo demonstrou que a construção do software atende à necessidade para o registro, monitoramento e a avaliação de feridas. As perspectivas são de que também possa ser utilizado para realização de simulação realística e treinamentos, além de possuir facilidade de acesso e manuseio

Validação e aplicabilidade do software Wounds Monitoring na avaliação e monitoramento de feridas

The aim was to validate an application software for recording, monitoring and evaluating wounds. Methodological, applied, quantitative study. The references adopted in this project were described in Standard No. 25010 of the International Organization for Standardization/International Electrotechnical Commission. The results were analyzed by a rule proposed by the Brazilian Association of Technical Norms in its Normative No. 14598-6. The feasibility of the system was evaluated through a test of the prototype, with the group of evaluators that was composed of two categories: specialists in information technology and nurses. In the evaluation of the nurses' expertise, the characteristic functional adequacy, reliability, usability, efficiency in performance, compatibility, and safety, obtained 100% of the responses in agreement. Professional IT experts also rated maintainability with 97.3% reliability with 84.9%, usability with 84.6% of the answers in agreement. The software was validated, allowing evaluating the technical quality and functional performance applied to the evaluation and monitoring of wounds.Objetivou-se validar um software aplicativo para registro, monitoramento e avaliação de feridas. Estudo metodológico, aplicado, quantitativo. As referências adotadas neste projeto foram descritas na Norma n.º 25010 da Internacional Organization for Standardization/International Electrotechnical Commission. Os resultados foram analisados por regra proposta pela Associação Brasileira de Normas Técnicas em sua Normativa n.º 14598-6. Foi avaliada a viabilidade do sistema através de um teste do protótipo, com o grupo de avaliadores que foi composto por duas categorias: especialistas em tecnologia da informação e enfermeiros. Na avaliação dos expertises enfermeiros, a característica adequação funcional confiabilidade, usabilidade, eficiência no desempenho, compatibilidade, e segurança, obtiveram 100% das respostas em acordo. Os expertises profissionais de informática também avaliaram manutenibilidade com 97,3% confiabilidade com 84,9%, usabilidade com 84,6% das respostas de acordo. O software foi validado permitindo avaliar a qualidade técnica e desempenho funcional aplicado à avaliação e monitoramento de feridas

Ocean Dumping of Containerized DDT Waste Was a Sloppy Process

Author Posting. © American Chemical Society, 2019. This article is posted here by permission of American Chemical Society for personal use, not for redistribution. The definitive version was published in Kivenson, V., Lemkau, K. L., Pizarro, O., Yoerger, D. R., Kaiser, C., Nelson, R. K., Carmichael, C., Paul, B. G., Reddy, C. M., & Valentine, D. L. (2019). Ocean Dumping of Containerized DDT Waste Was a Sloppy Process. Environmental Science and Technology (2019), doi:10.1021/acs.est.8b05859.Industrial-scale dumping of organic waste to the deep ocean was once common practice, leaving a legacy of chemical pollution for which a paucity of information exists. Using a nested approach with autonomous and remotely operated underwater vehicles, a dumpsite offshore California was surveyed and sampled. Discarded waste containers littered the site and structured the suboxic benthic environment. Dichlorodiphenyltrichloroethane (DDT) was reportedly dumped in the area, and sediment analysis revealed substantial variability in concentrations of p,p-DDT and its analogs, with a peak concentration of 257 μg g–1, ∼40 times greater than the highest level of surface sediment contamination at the nearby DDT Superfund site. The occurrence of a conspicuous hydrocarbon mixture suggests that multiple petroleum distillates, potentially used in DDT manufacture, contributed to the waste stream. Application of a two end-member mixing model with DDTs and polychlorinated biphenyls enabled source differentiation between shelf discharge versus containerized waste. Ocean dumping was found to be the major source of DDT to more than 3000 km2 of the region’s deep seafloor. These results reveal that ocean dumping of containerized DDT waste was inherently sloppy, with the contents readily breaching containment and leading to regional scale contamination of the deep benthos.This material is based upon work supported by the National Science Foundation Graduate Research Fellowship for V.K. under Grant No. 1650114. Expeditions AT-18-11 and AT-26-06 were funded by the NSF (OCE-0961725 and OCE-1046144). Any opinions, findings, and conclusions or recommendations expressed in this material are those of the author(s) and do not necessarily reflect the views of the National Science Foundation. We thank the captain and crew of the RV Atlantis, the pilots and crew of the ROV Jason, the crew of the AUV Sentry, the scientific party of the AT-18-11 and AT-26-06 expeditions, Justin Tran for assistance with the preparation of multibeam data, M. Indira Venkatesan for a helpful discussion of the NOAA datasets, and Nathan Dodder for advice on the procedure for compound identification

Synthesis and biological evaluation of novel MB327 analogs as resensitizers for desensitized nicotinic acetylcholine receptors after intoxication with nerve agents

Poisoning with organophosphorus compounds, which can lead to a cholinergic crisis due to the inhibition of acetylcholinesterase and the subsequent accumulation of acetylcholine (ACh) in the synaptic cleft, is a serious problem for which treatment options are currently insufficient. Our approach to broadening the therapeutic spectrum is to use agents that interact directly with desensitized nicotinic acetylcholine receptors (nAChRs) in order to induce functional recovery after ACh overstimulation. Although MB327, one of the most prominent compounds investigated in this context, has already shown positive properties in terms of muscle force recovery, this compound is not suitable for use as a therapeutic agent due to its insufficient potency. By means of in silico studies based on our recently presented allosteric binding pocket at the nAChR, i.e. the MB327-PAM-1 binding site, three promising MB327 analogs with a 4-aminopyridinium ion partial structure (PTM0056, PTM0062, and PTM0063) were identified. In this study, we present the synthesis and biological evaluation of a series of new analogs of the aforementioned compounds with a 4-aminopyridinium ion partial structure (PTM0064-PTM0072), as well as hydroxy-substituted analogs of MB327 (PTMD90–0012 and PTMD90–0015) designed to substitute entropically unfavorable water clusters identified during molecular dynamics simulations. The compounds were characterized in terms of their binding affinity towards the aforementioned binding site by applying the UNC0642 MS Binding Assays and in terms of their muscle force reactivation in rat diaphragm myography. More potent compounds were identified compared to MB327, as some of them showed a higher affinity towards MB327-PAM-1 and also a higher recovery of neuromuscular transmission at lower compound concentrations. To improve the treatment of organophosphate poisoning, direct targeting of nAChRs with appropriate compounds is a key step, and this study is an important contribution to this research

Role of β2-integrins for homing and neovascularization capacity of endothelial progenitor cells

The mechanisms of homing of endothelial progenitor cells (EPCs) to sites of ischemia are unclear. Here, we demonstrate that ex vivo–expanded EPCs as well as murine hematopoietic Sca-1+/Lin− progenitor cells express β2-integrins, which mediate the adhesion of EPCs to endothelial cell monolayers and their chemokine-induced transendothelial migration in vitro. In a murine model of hind limb ischemia, Sca-1+/Lin− hematopoietic progenitor cells from β2-integrin–deficient mice are less capable of homing to sites of ischemia and of improving neovascularization. Preactivation of the β2-integrins expressed on EPCs by activating antibodies augments the EPC-induced neovascularization in vivo. These results provide evidence for a novel function of β2-integrins in postnatal vasculogenesis

Management der Minderempfindlichkeit von Apfelwicklerstämmen gegenüber dem Apfelwicklergranulovirus

Das Apfelwicklergranulovirus (CpGV) ist effizientes biologisches Bekämpfungsmittel des Apfelwicklers mit großer Bedeutung im ökologischen und integrierten Kernobstbau. 2005 wurde erstmals eine Resistenz gegen CpGV in einzelnen Anlagen beobachtet. Um geeignete Hilfestellungen für den Obstbau zu entwickeln, wurden verschiedene Aspekte der CpGV-Resistenz aufgeklärt: 1. Bisher wurden mehr als 40 Apfelwicklerpopulationen in Europa mit CpGV-Resistenz gefunden. Sie kommen in Deutschland (26), Frankreich (2), Italien (6), Österreich (2), Schweiz (3), Niederlande (3) und Tschechien (1) vor. Vermutlich handelt es sich in allen Fällen um denselben Resistenztyp. 2. Die Resistenz wird durch einen ungewöhnlichen Vererbungsgang (einfaktoriell, Geschlechtschromosomal und mit einer konzentrationsabhängigen Dominanz) sehr effizient selektiert. 3. Der Resistenzmechanismus liegt in einer frühen Blockade der Virusvermehrung. Resistente Tiere zeigten keinen Fitnessnachteil gegenüber nicht resistenten Tieren in Laborexperimenten. 4. Neue CpGV-Isolate können Resistenz brechen. Viele der resistenzbrechenden Isolate wirken in resistenten Apfelwicklerlarven etwas langsamer als in anfälligen Larven. Dadurch ist auch bei Verwendung resistenzbrechender Tiere mit einem etwas erhöhten Schaden zu rechnen, solange bis die Apfelwicklerpopulation wieder auf ein niedriges Niveau reduziert wurde. Aus den Ergebnissen ergeben sich folgende Empfehlungen: 1. Betriebe ohne CpGV-Resistenz - dies ist die ganz überwiegende Zahl - können konventionelle CpGV-Präparate weiter verwenden. Sobald neue CpGV-Isolate zugelassen sind, sollte auch diese eingesetzt werden, um eine Selektion der bekannten Resistenz zu vermeiden. 2. Betriebe mit CpGV-Resistenz oder begründetem Resistenzverdacht sollten sofort neue resistenzbrechende Isolate verwenden. Diese stehen seit 2006 als Versuchspräparate zur Verfügung, ihre Zulassung wird erwartet. 3. Die Apfelwicklerbekämpfung muß auf eine möglichst breite Basis gestellt werden