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In this paper we consider how recent taonga puoro noho marae wānanga, in the Far North district of Aotearoa New Zealand, have much to offer the growth and development of bicultural relational engagement in arts education. The ngā toi Māori authors highlight their aspirations, as leaders of recent hui wānanga initiatives in Te Hiku o Te Ika (the region of the “tail of the fish” at the top of the North Island). This includes aspirations to recognise noho marae wānanga as important self-determining spaces that support the evolution of indigenous paradigms and the growth of music education from a distinctly Māori ontological foundation. Together we advocate for what Moana Jackson calls an “ethic of restoration” and argue that when priority is given to the recognition and support of Māori led educational initiatives, such as noho marae wānanga, generative and mutually beneficial bicultural relationships are better enabled. The authors revisit Te Mauri Pakeaka, an arts education marae wānanga initiative which ran through the 1980s, and consider hui wānanga for their potential to support new forms of bicultural relational engagement in arts education in the future.

Kaposi's sarcoma-associated herpesvirus (KSHV) is the etiological agent for Kaposi's sarcoma (KS), an HIV/AIDS-associated malignancy. Effective treatments against KS remain to be developed. The sugar analog 2-deoxy- d-glucose (2-DG) is an anticancer agent that is well-tolerated and safe in patients and was recently demonstrated to be a potent antiviral, including KSHV and severe acute respiratory syndrome coronavirus 2. Because 2-DG inhibits glycolysis and N-glycosylation, identifying its molecular targets is challenging. Here we compare the antiviral effect of 2-DG with 2-fluoro-deoxy- d-glucose, a glycolysis inhibitor, and 2-deoxy-fluoro- d-mannose (2-DFM), a specific N-glycosylation inhibitor. At doses similar to those clinically achievable with 2-DG, the three drugs impair KSHV replication and virion production in iSLK.219 cells via downregulation of viral structural glycoprotein expression (K8.1 and gB), being 2-DFM the most potent KSHV inhibitor. Consistently with the higher potency of 2-DFM, we found that d-mannose rescues KSHV glycoprotein synthesis and virus production, indicating that inhibition of N-glycosylation is the main antiviral target using d-mannose competition experiments. Suppression of N-glycosylation by the sugar drugs triggers ER stress. It activates the host unfolded protein response (UPR), counteracting KSHV-induced inhibition of the protein kinase R-like endoplasmic reticulum kinase branch, particularly activating transcription factor 4 and C/EBP homologous protein expression. Finally, we demonstrate that sugar analogs induce autophagy (a prosurvival mechanism) and, thus, inhibit viral replication playing a protective role against KSHV-induced cell death, further supporting their direct antiviral effect and potential therapeutic use. Our work identifies inhibition of N-glycosylation leading to ER stress and UPR as an antienveloped virus target and sugar analogs such as 2-DG and the newly identified 2-DFM as antiviral drugs.

BackgroundLike most major enzyme families, the M14 family of metallocarboxypeptidases (MCPs) contains a number of pseudoenzymes predicted to lack enzyme activity and with poorly characterized molecular function. The genome of the yeastSaccharomyces cerevisiaeencodes one member of the M14 MCP family, a pseudoenzyme named Ecm14 proposed to function in the extracellular matrix. In order to better understand the function of such pseudoenzymes, we studied the structure and function of Ecm14 inS. cerevisiae.ResultsA phylogenetic analysis of Ecm14 in fungi found it to be conserved throughout the ascomycete phylum, with a group of related pseudoenzymes found in basidiomycetes. To investigate the structure and function of this conserved protein, His6-tagged Ecm14 was overexpressed in Sf9 cells and purified. The prodomain of Ecm14 was cleaved in vivo and in vitro by endopeptidases, suggesting an activation mechanism; however, no activity was detectable using standard carboxypeptidase substrates. In order to determine the function of Ecm14 using an unbiased screen, we undertook a synthetic lethal assay. Upon screening approximately 27,000 yeast colonies, twenty-two putative synthetic lethal clones were identified. Further analysis showed many to be synthetic lethal with auxotrophic marker genes and requiring multiple mutations, suggesting that there are few, if any, singleS. cerevisiaegenes that present synthetic lethal interactions withecm14Δ.ConclusionsWe show in this study that Ecm14, although lacking detectable enzyme activity, is a conserved carboxypeptidase-like protein that is secreted from cells and is processed to a mature form by the action of an endopeptidase. Our study and datasets from other recent large-scale screens suggest a role for Ecm14 in processes such as vesicle-mediated transport and aggregate invasion, a fungal process that has been selected against in modern laboratory strains ofS. cerevisiae.

Roughly one-third of the human population is chronically infected with the intracellular single-celled parasite Toxoplasma gondii, but little is known about how this organism persists inside people. Previous research suggested that a parasite proteolytic enzyme, termed cathepsin protease L, is important for Toxoplasma persistence; however, it remained possible that other associated proteolytic enzymes could also be involved in the long-term survival of the parasite during infection. Here, we show that two proteolytic enzymes associated with cathepsin protease L play dispensable roles and are dependent on cathepsin L to reach maturity, which differs from the corresponding enzymes in humans. These findings establish a divergent hierarchy of proteases and help focus attention principally on cathepsin protease L as a potential target for interrupting Toxoplasma chronic infection., The lysosome-like vacuolar compartment (VAC) is a major site of proteolysis in the intracellular parasite Toxoplasma gondii. Previous studies have shown that genetic ablation of a VAC-residing cysteine protease, cathepsin protease L (CPL), resulted in the accumulation of undigested protein in the VAC and loss of parasite viability during the chronic stage of infection. However, since the maturation of another VAC localizing protease, cathepsin protease B (CPB), is dependent on CPL, it remained unknown whether these defects result directly from ablation of CPL or indirectly from a lack of CPB maturation. Likewise, although a previously described cathepsin D-like aspartyl protease 1 (ASP1) could also play a role in proteolysis, its definitive residence and function in the Toxoplasma endolysosomal system were not well defined. Here, we demonstrate that CPB is not necessary for protein turnover in the VAC and that CPB-deficient parasites have normal growth and viability in both the acute and chronic stages of infection. We also show that ASP1 depends on CPL for correct maturation, and it resides in the T. gondii VAC, where, similar to CPB, it plays a dispensable role in protein digestion. Taken together with previous work, our findings suggest that CPL is the dominant protease in a hierarchy of proteolytic enzymes within the VAC. This unusual lack of redundancy for CPL in T. gondii makes it a single exploitable target for disrupting chronic toxoplasmosis. IMPORTANCE Roughly one-third of the human population is chronically infected with the intracellular single-celled parasite Toxoplasma gondii, but little is known about how this organism persists inside people. Previous research suggested that a parasite proteolytic enzyme, termed cathepsin protease L, is important for Toxoplasma persistence; however, it remained possible that other associated proteolytic enzymes could also be involved in the long-term survival of the parasite during infection. Here, we show that two proteolytic enzymes associated with cathepsin protease L play dispensable roles and are dependent on cathepsin L to reach maturity, which differs from the corresponding enzymes in humans. These findings establish a divergent hierarchy of proteases and help focus attention principally on cathepsin protease L as a potential target for interrupting Toxoplasma chronic infection.

Additional file 6. Original unmodified western blots.

Additional file 3: Supporting information. Table S1. Whole-genome sequencing using the Illumina MiniSeq® platform. Table S2. Plasmids constructed and used in this study. Figure S1. Expression and purification of Ecm14. Figure S2. Edman degradation spectra. Figure S3. Large-scale screens identifying phenotypes for ECM14 mutants are not reproducible. Figure S4. Map of pSLS1-ECM14. Figure S5. The effect of EMS exposure time on yeast cell viability. Figure S6. Putative synthetic lethal mutants have reduced fitness on minimal media.

Predicting how communities respond to multiple, potentially interacting chemical stressors is inherently difficult because community structure and dynamics, the chemical properties of contaminants, and biological-chemical interactions vary with environmental conditions. Using a field experiment conducted in Antarctica, we tested whether 3 phyla of benthic soft-sediment marine invertebrates—annelids, arthropods, and echinoderms —respond differently to 2 common forms of contamination, organic enrichment and toxic contamination. Based on life history strategies and physiological tolerances to contaminants, we hypothesized that the principal responses of the 3 phyla would be: (1) enhanced abundance of annelids in organically enrichment sediments and (2) decreased abundance of arthropods and echinoderms in toxic metal contamination. Sediment treatments were established in the field experiment with an orthogonal combination of 3 levels of total organic carbon (TOC; 0, 1, and 2% by weight) and copper (Cu; 0, 100, and 500 µg Cu g -1 sedi- ment), and colonization patterns were observed after 1 yr. Densities of annelids (mainly polychaetes) increased with TOC across all levels of Cu. Arthropods and echinoderms decreased with Cu, but responded variably to TOC, based largely on differences in habitat preferences exhibited by epifau- nal and infaunal species. Small subsurface arthropod species (amphipods, isopods, cumaceans, and ostracods) decreased in high organic loading, due to induction of and exposure to hypoxia and hydro- gen sulfide, but large surface deposit-feeding echinoderms (asteroids and echinoids) responded positively to increased carbon food supply. We present a predictive model based on assessment of benthic community structure conducted at the taxonomic level of phyla that could be used to link cause and effect for multiple chemical stressors in marine ecosystems.