Vps34p differentially regulates endocytosis from the apical and basolateral domains in polarized hepatic cells

Using a microinjection approach to study apical plasma membrane protein trafficking in hepatic cells, we found that specific inhibition of Vps34p, a class III phosphoinositide 3 (PI-3) kinase, nearly perfectly recapitulated the defects we reported for wortmannin-treated cells (Tuma, P.L., C.M. Finnegan, J.-H Yi, and A.L. Hubbard. 1999. J. Cell Biol. 145:1089–1102). Both wortmannin and injection of inhibitory Vps34p antibodies led to the accumulation of resident apical proteins in enlarged prelysosomes, whereas transcytosing apical proteins and recycling basolateral receptors transiently accumulated in basolateral early endosomes. To understand how the Vps34p catalytic product, PI(3)P, was differentially regulating endocytosis from the two domains, we examined the PI(3)P binding protein early endosomal antigen 1 (EEA1). We determined that EEA1 distributed to two biochemically distinct endosomal populations: basolateral early endosomes and subapical endosomes. Both contained rab5, although the latter also contained late endosomal markers but was distinct from the transcytotic intermediate, the subapical compartment. When PI(3)P was depleted, EEA1 dissociated from basolateral endosomes, whereas it remained on subapical endosomes. From these results, we conclude that PI(3)P, via EEA1, regulates early steps in endocytosis from the basolateral surface in polarized WIF-B cells. However, PI(3)P must use different machinery in its regulation of the apical endocytic pathway, since later steps are affected by Vps34p inhibition

Generalist Primary School Teachers’ Preferences for Becoming Subject Matter Specialists

Traditionally Australian primary school teachers have been viewed as generalists responsible for instruction across all content areas. Adopting self-determination theory as a lens, the aim of the study was to explore the extent to which generalist primary school teachers are interested in becoming subject matter specialists. Questionnaire data were collected from 104 early years primary school teachers. Findings suggest that two-thirds of these generalist teachers expressed an interest in specialising in either English, mathematics, and to a far lesser extent, science, such that they would be responsible for exclusively teaching this subject. Preferences for specialisation were based on teachers’ self-perceived content and pedagogical expertise and/ or their enjoyment of teaching in this content area. By contrast, the one-third of teachers who would choose to remain generalists referred to the value in a variety of teaching experiences, teaching from a whole child perspective and content integration. Implications for educational policy are discussed

The Role of Differential Ablation and Dynamic Detachment in Driving Accelerating Mass Loss From a Debris-Covered Himalayan Glacier

Sustained mass loss from Himalayan glaciers is causing supraglacial debris to expand and thicken, with the expectation that thicker debris will suppress ablation and extend glacier longevity. However, debris-covered glaciers are losing mass at similar rates to clean-ice glaciers in High Mountain Asia. This rapid mass loss is attributed to the combined effects of; (a) low or reversed mass balance gradients across debris-covered glacier tongues, (b) differential ablation processes that locally enhance ablation within the debris-covered section of the glacier, for example, at ice cliffs and supraglacial ponds, and (c) a decrease in ice flux from the accumulation area in response to climatic warming. Adding meter-scale spatial variations in supraglacial debris thickness to an ice-flow model of Khumbu Glacier, Nepal, increased mass loss by 47% relative to simulations assuming a continuous debris layer over a 31-year period (1984–2015 CE) but overestimated the reduction in ice flux. Therefore, we investigated if simulating the effects of dynamic detachment of the upper active glacier from the debris-covered tongue would give a better representation of glacier behavior, as suggested by observations of change in glacier dynamics and structure indicating that this process occurred during the last 100 years. Observed glacier change was reproduced more reliably in simulations of the active, rather than entire, glacier extent, indicating that Khumbu Glacier has passed a dynamic tipping point by dynamically detaching from the heavily debris-covered tongue that contains 20% of the former ice volume

Surface and subsurface hydrology of debris-covered Khumbu Glacier, Nepal, revealed by dye tracing

While the supraglacial hydrology of debris-covered glaciers is relatively well studied, almost nothing is known about how water is transported beneath the glacier surface. Here, we report the results of sixteen fluorescent dye tracing experiments conducted in April–May 2018 over the lowermost 7 km of the high-elevation, debris-covered Khumbu Glacier, Nepal, to characterise the glacier's surface and subsurface drainage system. Dye breakthroughs indicated a likely highly sinuous and channelised subsurface hydrological system draining water from the upper part of the ablation area. This flowpath was distinct from the linked chain of supraglacial ponds present along much of the glacier's lower ablation area, through which water flow was extremely slow (∼0.003 m s−1), likely reflecting the study's timing during the pre-monsoon period. Subsurface drainage pathways emerged at the glacier surface close to the terminus, and flowed into small near-surface englacial reservoirs that typically delayed meltwater transit by several hours. We observed rapid pathway changes resulting from surface collapse, indicating a further distinctive aspect of the drainage of debris-covered glaciers. We conclude that the surface and subsurface drainage of Khumbu Glacier is both distinctive and dynamic, and argue that further investigation is needed to refine the characterisation and test its regional applicability to better understand future Himalayan debris-covered glacier meltwater delivery to downstream areas

Exploration of piperidine 3D fragment chemical space : synthesis and 3D shape analysis of fragments derived from 20 regio- and diastereoisomers of methyl substituted pipecolinates

Fragment-based drug discovery is now widely adopted for lead generation in the pharmaceutical industry. However, fragment screening collections are often predominantly populated with flat, 2D molecules. Herein, we report the synthesis of piperidine-based 3D fragment building blocks - 20 regio- and diastereoisomers of methyl substituted pipecolinates using simple and general synthetic methods. cis-Piperidines, accessed through a pyridine hydrogenation were transformed into their trans-diastereoisomers using conformational control and unified reaction conditions. Additionally, diastereoselective lithiation/trapping was utilised to access trans-piperidines. Analysis of a virtual library of fragments derived from the 20 cis- and trans-disubstituted piperidines showed that it consisted of 3D molecules with suitable molecular properties to be used in fragment-based drug discovery programs

Supraglacial ponds regulate runoff from Himalayan debris-covered glaciers

Meltwater and runoff from glaciers in High Mountain Asia is a vital freshwater resource for one fifth of the Earth's population. Between 13% and 36% of the region's glacierized areas exhibit surface debris cover and associated supraglacial ponds whose hydrological buffering roles remain unconstrained. We present a high-resolution meltwater hydrograph from the extensively debris-covered Khumbu Glacier, Nepal, spanning a seven-month period in 2014. Supraglacial ponds and accompanying debris cover modulate proglacial discharge by acting as transient and evolving reservoirs. Diurnally, the supraglacial pond system may store >23% of observed mean daily discharge, with mean recession constants ranging from 31 to 108 hours. Given projections of increased debris-cover and supraglacial pond extent across High Mountain Asia, we conclude that runoff regimes may become progressively buffered by the presence of supraglacial reservoirs. Incorporation of these processes is critical to improve predictions of the region's freshwater resource availability and cascading environmental effects downstream