RNAi Trigger Delivery into Anopheles gambiae Pupae

Citation: Regna, K., Harrison, R. M., Heyse, S. A., Chiles, T. C., Michel, K., & Muskavitch, M. A. T. (2016). RNAi Trigger Delivery into Anopheles gambiae Pupae. Jove-Journal of Visualized Experiments(109), 9. doi:10.3791/53738RNA interference (RNAi), a naturally occurring phenomenon in eukaryotic organisms, is an extremely valuable tool that can be utilized in the laboratory for functional genomic studies. The ability to knockdown individual genes selectively via this reverse genetic technique has allowed many researchers to rapidly uncover the biological roles of numerous genes within many organisms, by evaluation of loss-of-function phenotypes. In the major human malaria vector Anopheles gambiae, the predominant method used to reduce the function of targeted genes involves injection of double-stranded (dsRNA) into the hemocoel of the adult mosquito. While this method has been successful, gene knockdown in adults excludes the functional assessment of genes that are expressed and potentially play roles during pre-adult stages, as well as genes that are expressed in limited numbers of cells in adult mosquitoes. We describe a method for the injection of Serine Protease Inhibitor 2 (SRPN2) dsRNA during the early pupal stage and validate SRPN2 protein knockdown by observing decreased target protein levels and the formation of melanotic pseudo-tumors in SRPN2 knockdown adult mosquitoes. This evident phenotype has been described previously for adult stage knockdown of SRPN2 function, and we have recapitulated this adult phenotype by SRPN2 knockdown initiated during pupal development. When used in conjunction with a dye-labeled dsRNA solution, this technique enables easy visualization by simple light microscopy of injection quality and distribution of dsRNA in the hemocoel

PHIDL: Python CAD layout and geometry creation for nanolithography

Computer-aided design (CAD) has become a critical element in the creation of nanopatterned structures and devices. In particular, with the increased adoption of easy-to-learn programming languages like Python there has been a significant rise in the amount of lithographic geometries generated through scripting and programming. However, there are currently unaddressed gaps in usability for open-source CAD tools -- especially those in the GDSII design space -- that prevent wider adoption by scientists and students who might otherwise benefit from scripted design. For example, constructing relations between adjacent geometries is often much more difficult than necessary -- spacing a resonator structure a few micrometers from a readout structure often requires manually-coding the placement arithmetic. While inconveniences like this can be overcome by writing custom functions, they are often significant barriers to entry for new users or those less familiar with programming. To help streamline the design process and reduce barrier to entry for scripting designs, we have developed PHIDL, an open-source GDSII-based CAD tool for Python 2 and 3

Resident Gut Microbiota Mediates Exercise Capacity and Tissue Metabolomes in Mice

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Metabolomic Shifts Associated with Heat Stress in Coral Holobionts

Understanding the response of the coral holobiont to environmental change is crucial to inform conservation efforts. The most pressing problem is “coral bleaching,” usually precipitated by prolonged thermal stress. We used untargeted, polar metabolite profiling to investigate the physiological response of the coral species Montipora capitata and Pocillopora acuta to heat stress. Our goal was to identify diagnostic markers present early in the bleaching response. From the untargeted UHPLC-MS data, a variety of co-regulated dipeptides were found that have the highest differential accumulation in both species. The structures of four dipeptides were determined and showed differential accumulation in symbiotic and aposymbiotic (alga-free) populations of the sea anemone Aiptasia (Exaiptasia pallida), suggesting the deep evolutionary origins of these dipeptides and their involvement in symbiosis. These and other metabolites may be used as diagnostic markers for thermal stress in wild coral

Digital characterization of particle tracks for microdosimetry

Work is in progress to develop a digital approach to microdosimetry and to construct a prototype instrument to obtain digital information about charged-particle tracks. The objective of such a device is to measure the numbers of electrons produced in various subvolumes of a chamber gas along a particle's path. This paper describes results of Monte Carlo calculations of charged-particle tracks in a cubical time-projection ionization chamber containing methane. Results are presented to show the effects of electron diffusion during charge collection. The calculations indicate that the optimum ratio of field strength and pressure is about 0.6 volts cm/sup -1/ torr/sup -1/. Examples of proton, carbon-ion, and electron tracks are shown. 2 refs., 6 figs

Evaluation of the Workplace Environment in the UK, and the Impact on Users’ Levels of Stimulation

The purpose of this study is to evaluate a number of recently completed workplaces in the UK. The first aim is to assess the impact of various aspects of the workplace environment on users’ levels of stimulation. The body of previous research undertaken into the workplace environment, identified the aspects to be investigated. Samples of employees from the sixteen businesses were surveyed to determine their perceptions of the workplaces. The results were entered into a regression analysis, and the most significant predictors of perceived stimulation identified. The data also revealed a dramatic reduction in staff arousal levels from mornings to afternoons. Thus, there is a second aim to determine whether changes to significant aspects of the workplace environment during the day can counteract the reduction in users’ stimulation. Two further workplaces were studied to enable changes to be made over a 12-week period. A sample of employees completed questionnaires, and semi-structured interviews revealed the reasons behind the results. It was found that provision of artwork, personal control of temperature and ventilation and regular breaks were the most significant contributions to increasing stimulation after lunch; while user choice of layout, and design and décor of workspaces and break areas, were the most significant aspects at design stage

Announcing the Minderoo - Monaco Commission on Plastics and Human Health.

© The Author(s), 2022. This article is distributed under the terms of the Creative Commons Attribution License. The definitive version was published in Landrigan, P., Raps, H., Symeonides, C., Chiles, T., Cropper, M., Enck, J., Hahn, M., Hixson, R., Kumar, P., Mustapha, A., Park, Y., Spring, M., Stegeman, J., Thompson, R., Wang, Z., Wolff, M., Yousuf, A., & Dunlop, S. Announcing the Minderoo – Monaco Commission on Plastics and Human Health. Annals of Global Health, 88(1), (2022): 73, https://doi.org/10.5334/aogh.3916.Plastic is the signature material of our age. In the 75 years since large-scale production began in the aftermath of World War II, plastic has transformed our world, supported many of the most significant advances of modern civilization, and enabled breakthroughs in virtually every field of human endeavor. But plastic also poses great and growing dangers to human health and the environment, harms that fall disproportionately on the world’s poorest and most vulnerable populations. The extent and magnitude of these dangers are only beginning to be understood.The funding is from the Minderoo Foundation, the Centre Scientifique de Monaco, and the Prince Albert II of Monaco Foundation

Thickening of the pulmonary artery wall in acute intramural hematoma of the ascending aorta

BACKGROUND: The occurrence of pulmonary artery obstruction in the course of acute aortic dissection is an unusual complication. The mechanism implicated is the rupture of the outer layer of the aorta and the subsequent hemorrhage into the adventitia of the pulmonary artery that causes its wall thickening and, at times, produces extrinsic obstruction of the vessel. There are no reports of this complication in acute intramural hematoma. CASE PRESENTATION: An 87-year-old woman was admitted to the hospital in shock after having had severe chest pain followed by syncope. An urgent transesophageal echocardiogram revealed the presence of acute intramural hematoma, no evidence of aortic dissection, severe pericardial effusion with cardiac tamponade, and periaortic hematoma that involved the pulmonary artery generating circumferential wall thickening of its trunk and right branch with no evidence of flow obstruction. Urgent surgery was performed but the patient died in the operating room. The post mortem examination, in the operating room, confirmed that there was an extensive hematoma around the aorta and beneath the adventitial layer of the pulmonary artery, with no evidence of flow obstruction. CONCLUSION: This is the first time that this rare complication is reported in the scenario of acute intramural hematoma and with the transesophageal echocardiogram as the diagnostic tool

Caracterização e análise financeira do processo de extração de óleo de polpa de pequi no Cerrado brasileiro.

Resumo ? O presente estudo realizou caracterização e análise financeira do processo de extração de óleo de polpa de pequi, uma atividade secular de populações tradicionais do Território da Cidadania Alto Rio Pardo (MG). O estudo considerou os custos de insumos e de trabalho das sete etapas do processo produtivo, além da consideração dos riscos de contaminação de ferro no cozimento e na despolpa, mas desconsiderou a comparação entre o lucro obtido por hora de trabalho e o valor da diária de trabalho local, afinal, as relações capitalistas de produção não estão plenamente inseridas na realidade local. Entre as sete etapas analisadas, três apresentaram variações, sendo elas, a colheita (duas variáveis), cozimento (seis variáveis) e despolpa (quatro variáveis), resultando em 48 cenários hipotéticos de processos de extração. Entre eles, quatro combinações são recomendadas ao contexto de vida das famílias rurais locais (colheita na unidade de produção; cozimento no tacho de cobre ou na panela de alumínio; despolpa no engenho manual ou no engenho a motor) e uma combinação é mais adequada ao contexto dos processos agroindustriais coletivos presentes na região (colheita na unidade de produção; cozimento na panela de inox; despolpa no engenho a motor)