Genetic Engineering of the Rock Inhabitant Knufia petricola Provides Insight Into the Biology of Extremotolerant Black Fungi

Black microcolonial fungi (Ascomycetes from Arthonio-, Dothideo-, and Eurotiomycetes) are stress-tolerant and persistent dwellers of natural and anthropogenic extreme habitats. They exhibit slow yeast-like or meristematic growth, do not form specialized reproduction structures and accumulate the black pigment 1,8-dihydroxynaphthalene (DHN) melanin in the multilayered cell walls. To understand how black fungi live, survive, colonize mineral substrates, and interact with phototrophs genetic methods are needed to test these functions and interactions. We chose the rock inhabitant Knufia petricola of the Chaetothyriales as a model for developing methods for genetic manipulation. Here, we report on the expansion of the genetic toolkit by more efficient multiplex CRISPR/Cas9 using a plasmid-based system for expression of Cas9 and multiple sgRNAs and the implementation of the three resistance selection markers genR (geneticin/nptII), baR (glufosinate/bar), and suR (chlorimuron ethyl/sur). The targeted integration of expression constructs by replacement of essential genes for pigment synthesis allows for an additional color screening of the transformants. The black-pink screening due to the elimination of pks1 (melanin) was applied for promoter studies using GFP fluorescence as reporter. The black-white screening due to the concurrent elimination of pks1 and phs1 (carotenoids) allows to identify transformants that contain the two expression constructs for co-localization or bimolecular fluorescence complementation (BiFC) studies. The co-localization and interaction of the two K. petricola White Collar orthologs were demonstrated. Two intergenic regions (igr1, igr2) were identified in which expression constructs can be inserted without causing obvious phenotypes. Plasmids of the pNXR-XXX series and new compatible entry plasmids were used for fast and easy generation of expression constructs and are suitable for a broad implementation in other fungi. This variety of genetic tools is opening a completely new perspective for mechanistic and very detailed study of expression, functioning and regulation of the genes/proteins encoded by the genomes of black fungi

Correlates of Executive Dysfunction in HIV

Despite advancements in combination antiretroviral therapy rates of mild neurocognitive impairment remain elevated amongst persons living with HIV (PLWH). Concomitant with the greater rate of survival into old age PLWH show accelerated cognitive Aging in multiple domains. Executive function is among the domains where early onset of decline is most evident. This chapter will discuss what are known correlates of executive dysfunction in PLWH in this era of antiretroviral therapy. The goal of this chapter is to expand on what is known regarding HIV disease severity and executive dysfunction to provide a more nuanced look at how other central and peripheral biomarkers, psychosocial and behavioral risk factors relate to deficits in inhibition, updating and working memory, cognitive set-shifting, and mental flexibility

Productivity and Linkages of the Food Web of the Southern Region of the Western Antarctic Peninsula Continental Shelf

The productivity and linkages in the food web of the southern region of the west Antarctic Peninsula continental shelf were investigated using a multi-trophic level mass balance model. Data collected during the Southern Ocean Global Ocean Ecosystem Dynamics field program were combined with data from the literature on the abundance and diet composition of zooplankton, fish, seabirds and marine mammals to calculate energy flows in the food web and to infer the overall food web structure at the annual level. Sensitivity analyses investigated the effects of variability in growth and biomass of Antarctic krill (Euphausia superba) and in the biomass of Antarctic krill predators on the structure and energy fluxes in the food web. Scenario simulations provided insights into the potential responses of the food web to a reduced contribution of large phytoplankton (diatom) production to total primary production, and to reduced consumption of primary production by Antarctic krill and mesozooplankton coincident with increased consumption by microzooplankton and salps. Model-derived estimates of primary production were 187–207 g C m−2 y−1, which are consistent with observed values (47–351 g C m−2 y−1). Simulations showed that Antarctic krill provide the majority of energy needed to sustain seabird and marine mammal production, thereby exerting a bottom-up control on higher trophic level predators. Energy transfer to top predators via mesozooplanton was a less efficient pathway, and salps were a production loss pathway because little of the primary production they consumed was passed to higher trophic levels. Increased predominance of small phytoplankton (nanoflagellates and cryptophytes) reduced the production of Antarctic krill and of its predators, including seabirds and seals

Erratum to: Guidelines for the use and interpretation of assays for monitoring autophagy (3rd edition) (Autophagy, 12, 1, 1-222, 10.1080/15548627.2015.1100356

non present