Guidelines for the use and interpretation of assays for monitoring autophagy

In 2008 we published the first set of guidelines for standardizing research in autophagy. Since then, research on this topic has continued to accelerate, and many new scientists have entered the field. Our knowledge base and relevant new technologies have also been expanding. Accordingly, it is important to update these guidelines for monitoring autophagy in different organisms. Various reviews have described the range of assays that have been used for this purpose. Nevertheless, there continues to be confusion regarding acceptable methods to measure autophagy, especially in multicellular eukaryotes. A key point that needs to be emphasized is that there is a difference between measurements that monitor the numbers or volume of autophagic elements (e.g., autophagosomes or autolysosomes) at any stage of the autophagic process vs. those that measure flux through the autophagy pathway (i.e., the complete process); thus, a block in macroautophagy that results in autophagosome accumulation needs to be differentiated from stimuli that result in increased autophagic activity, defined as increased autophagy induction coupled with increased delivery to, and degradation within, lysosomes (in most higher eukaryotes and some protists such as Dictyostelium) or the vacuole (in plants and fungi). In other words, it is especially important that investigators new to the field understand that the appearance of more autophagosomes does not necessarily equate with more autophagy. In fact, in many cases, autophagosomes accumulate because of a block in trafficking to lysosomes without a concomitant change in autophagosome biogenesis, whereas an increase in autolysosomes may reflect a reduction in degradative activity. Here, we present a set of guidelines for the selection and interpretation of methods for use by investigators who aim to examine macroautophagy and related processes, as well as for reviewers who need to provide realistic and reasonable critiques of papers that are focused on these processes. These guidelines are not meant to be a formulaic set of rules, because the appropriate assays depend in part on the question being asked and the system being used. In addition, we emphasize that no individual assay is guaranteed to be the most appropriate one in every situation, and we strongly recommend the use of multiple assays to monitor autophagy. In these guidelines, we consider these various methods of assessing autophagy and what information can, or cannot, be obtained from them. Finally, by discussing the merits and limits of particular autophagy assays, we hope to encourage technical innovation in the field.

Analyzing the effects of vertical acceleration and deceleration in multi-phase, debris-flow sediment-column experiments / by Christine Marie Williams.

Water-saturated debris flows are among some of the most destructive mass movements. Their complex nature presents a challenge for quantitative description and modeling. In order to improve understanding of the dynamics of these flows, it is important to seek a simplified dynamic system underlying their behavior. Models currently in use to describe the motion of debris flows employ depth-averaged equations of motion, typically assuming negligible effects from vertical acceleration. However, in many cases debris flows experience significant vertical acceleration as they move across irregular surfaces, and it has been proposed that friction associated with vertical forces and liquefaction merit inclusion in any comprehensive mechanical model. The intent of this work is to determine the effect of vertical acceleration through a series of laboratory experiments designed to simulate debris flows, testing a recent model for debris flows experimentally. In the experiments, a mass of water-saturated sediment is released suddenly from a holding container, and parameters including rate of collapse, pore-fluid pressure, and bed load are monitored. Experiments are simplified to axial geometry so that variables act solely in the vertical dimension. Steady state equations to infer motion of the moving sediment mass are not sufficient to model accurately the independent solid and fluid constituents in these experiments. The model developed in this work more accurately predicts the bed-normal stress of a saturated sediment mass in motion and illustrates the importance of acceleration and deceleration.

The Other Side of the Doors: Butte & Beyond -- Congressman Pat Williams “In the Crucible of Change”

Pat Williams emerged from the Mining City of Butte, Montana with a sense of grassroots, people-oriented politics. His inherent belief in the power of ordinary citizens carried him through the Montana Legislature and into Congress for a record-setting period. The accomplishments of his long career partially obscured his innate progressive and populist instinct that is reflective of the period of “in the Crucible of Change.” This film addresses Pat’s early years when his progressive instincts and activities resulted in pushback from the giant Anaconda Company which had held Montana hostage for 75 years. Pat is joined for part of the film by former campaign staffer, and now prominent media consultant, Michael Fenenbock for reflections on Pat’s 1978 “Door-to-Door to Congress” campaign, which demonstrated the power of his belief in the people on the other side of the doors. Pat Williams (b. 1937) rose from teaching grade school in his hometown of Butte, MT, to serving for the longest number of consecutive terms (9 terms, 18 years) in the US House of Representatives of anyone in Montana history. Pat was a member of the National Guard and attended UM in Missoula and William Jewel College, graduating from the University of Denver. Pat also served in the Montana legislature for 2 terms (1966 & 1968 elections). In 1969. Pat helped his legislative seat-mate John Melcher get elected as Montana’s Eastern District Congressman in the Special Election that June. Pat went to Washington DC as Melcher’s Executive Assistant. Upon returning to Montana, Pat headed up the Montana offices of the innovative Mountain Plains Family Education Program. In 1974, Pat ran unsuccessfully for Montana’s Western District Congressional seat in a three-way race with former Congressman Arnold Olsen and state Legislator Max Baucus. After the drafting and passage of the 1972 Montana Constitution, Pat was named a member of Montana’s first-ever Reapportionment Commission. In 1978 he successfully ran for Congress, conducting a massive grass-roots door-to-door campaign of 1½ years, reaching 50,000 doors. In a hotly contested 6-way Democratic primary, Pat won going away and also handily won the general election. Pat served in Congress from January, 1979 until January of 1997, 14 years representing the Western District and 4 years representing the entire state. Upon his retirement from Congress, in 1997 Williams returned to Montana where has been an instructor at the University of Montana and Senior Fellow and Regional Policy Associate at the Center for the Rocky Mountain West. He is a former member of the Montana Board of Regents and serves on a number of national education-related boards. In Congress Pat was a Deputy Whip of the U.S. House of Representatives and sat on committees on: Budget, Natural Resources, Education and Labor, and Agriculture. Pat’s leadership helped pass trailblazing legislation to assist hard-working middle-class families and ensure opportunities for every child. Pat’s fingerprints are on many pieces of important legislation, including the College Middle Income Assistance Act, the Family and Medical Leave Act, the Toddlers and Childhood Disability Act, the Library Services and Construction Act, and the Museum Services Act. Pat successfully sponsored the Lee Metcalf Wilderness Area and the Rattlesnake Wilderness area, helped save the Bob Marshall Wilderness from oil and gas exploration, and helped ban geothermal energy drilling near the borders of Yellowstone National Park. As Chairman of The Post-Secondary Education Committee, he protected the National Endowment for the Arts from elimination, a remarkable undertaking during a very trying time for the Agency. Pat worked tirelessly with Tribal College Leaders to build Montana’s seven Tribal Colleges. He was also responsible for the legislation that created The American Conservation Corps, which became the Corporation for National Service, giving thousands of America’s young people a chance to serve their country and pursue higher education. Pat lives in Missoula with his wife Carol Griffith Williams, former Montana Senate Majority Leader. They have three children and five grandchildren.

Evaluation of Cage Designs and Feeding Regimes for Honey Bee (Hymenoptera: Apidae) Laboratory Experiments

The aim of this study was to improve cage systems for maintaining adult honey bee (Apis mellifera L.) workers under in vitro laboratory conditions. To achieve this goal, we experimentally evaluated the impact of different cages, developed by scientists of the international research network COLOSS (Prevention of honey bee COlony LOSSes), on the physiology and survival of honey bees. We identified three cages that promoted good survival of honey bees. The bees from cages that exhibited greater survival had relatively lower titers of deformed wing virus, suggesting that deformed wing virus is a significant marker reflecting stress level and health status of the host. We also determined that a leak- and drip-proof feeder was an integral part of a cage system and a feeder modified from a 20-ml plastic syringe displayed the best result in providing steady food supply to bees. Finally, we also demonstrated that the addition of protein to the bees' diet could significantly increase the level ofvitellogenin gene expression and improve bees' survival. This international collaborative study represents a critical step toward improvement of cage designs and feeding regimes for honey bee laboratory experiments.

Harriett Jay, Kniestück

Signatur des Originals: S 36/F08900

Anna Williams, Ganzfigurbildnis, Profil

Signatur des Originals: S 36/F10278

Anna Williams, Rollenbild, Kniestück

Signatur des Originals: S 36/F11929

Evaluación de las características físico-químicas y biológicas en dos suelos superficiales cultivados con pera (Pyrus communis L.) cv. Williams bajo manejo convencional

El objetivo del trabajo fue identificar las características físico-químicas y biológicas en dos suelos superficiales fertilizados con nitrógeno y enmienda orgánica en el Alto Valle de Río Negro (huertos H1 y H2). En ambos huertos se aplicó fertilizante nitrogenado durante las temporadas 2008-2009 y 2009-2010 y en H2 se aplicó estiércol de pollo. Se extrajeron muestras de suelos en primavera y otoño y se determinó: carbono orgánico total, conductividad eléctrica, cationes de intercambio, relación de adsorción de sodio y nitratos, respiración microbiana, carbono de la biomasa microbiana, actividad de la deshidrogenasa y el índice de mineralización. La concentración de carbono orgánico total, potasio y nitrógeno fueron adecuadas para la producción de pera. El comportamiento de las variables biológicas fue diferente en los huertos. En H1 fueron mayores en primavera, en ambas temporadas y el índice de mineralización fue ligeramente superior a 1 en otoño, indicando equilibrio entre la mineralización y la humificación del carbono. En H2 las mediciones biológicas fueron similares entre las estaciones como consecuencia de realizar fertilización nitrogenada (N) en primavera y en otoño. La enmienda orgánica no reflejó un aumento de la actividad biológica en primavera. La actividad microbiana y enzimática en H1 y H2 fue sensible a los cambios que ocurrieron en los suelos.

Sandra Visser y Thomas Williams, Anselm, (Great Medieval Thinker), Oxford University Press, Oxford, 2009; 303 p. ISBN 978-0-19-530939-3

Fil: Peretó Rivas, Rubén.