Regulatory Elements within the Prodomain of Falcipain-2, a Cysteine Protease of the Malaria Parasite Plasmodium falciparum

Falcipain-2, a papain family cysteine protease of the malaria parasite Plasmodium falciparum, plays a key role in parasite hydrolysis of hemoglobin and is a potential chemotherapeutic target. As with many proteases, falcipain-2 is synthesized as a zymogen, and the prodomain inhibits activity of the mature enzyme. To investigate the mechanism of regulation of falcipain-2 by its prodomain, we expressed constructs encoding different portions of the prodomain and tested their ability to inhibit recombinant mature falcipain-2. We identified a C-terminal segment (Leu155–Asp243) of the prodomain, including two motifs (ERFNIN and GNFD) that are conserved in cathepsin L sub-family papain family proteases, as the mediator of prodomain inhibitory activity. Circular dichroism analysis showed that the prodomain including the C-terminal segment, but not constructs lacking this segment, was rich in secondary structure, suggesting that the segment plays a crucial role in protein folding. The falcipain-2 prodomain also efficiently inhibited other papain family proteases, including cathepsin K, cathepsin L, cathepsin B, and cruzain, but it did not inhibit cathepsin C or tested proteases of other classes. A structural model of pro-falcipain-2 was constructed by homology modeling based on crystallographic structures of mature falcipain-2, procathepsin K, procathepsin L, and procaricain, offering insights into the nature of the interaction between the prodomain and mature domain of falcipain-2 as well as into the broad specificity of inhibitory activity of the falcipain-2 prodomain

The Life of John Wishart (1850–1926): Study of an Academic Surgical Career Prior to the Flexner Report

BACKGROUND: The 1910 Flexner Report on Medical Education in the United States and Canada is often taken as the point when medical schools in North America took on their modern form. However, many fundamental advances in surgery, such as anesthesia and asepsis, predated the report by decades. To understand the contribution of educators in this earlier period, we investigated the forgotten career of John Wishart, founding Professor of Surgery at Western University, London Ontario. METHODS: Archives at the University of Western Ontario, University of Toronto, London City Library, and Wellington County Museum were searched for material about Wishart and his times. RESULTS: A fragmented biography can be assembled from family notes and obituaries with the help of contemporary documents compiled by early 20th century medical school historians. Wishart assisted Abraham Groves in the first reported operation for which aseptic technique was used (1874). He was considered locally to perform pioneering surgery, including an appendectomy in 1886. Wishart was a founding member of the medical faculty at Western University in 1881, initially as Demonstrator of Anatomy and subsequently as its first Professor of Clinical Surgery, which post he held until 1910. Comprehensive notes from his undergraduate lectures demonstrate his teaching style, which mixed organized didacticism with practical advice. The role of the Flexner review in the termination of his professorship is hinted at in minutes of Faculty of Medicine meetings. Wishart was a foundation fellow of the American College of Surgeons and a founding physician of London\u27s Catholic hospital, St. Joseph\u27s, despite his own Protestant background. CONCLUSIONS: Wishart\u27s career comprised all the elements of modern academic surgery, including pioneering service, research, and teaching. Surgery at Western owes as much to Wishart as it does to university reorganization in response to the Flexner report. PMID: 2227097

Worry about crime in Europe: A model-based small area estimation from the European Social Survey

© The Author(s) 2019. Worry about crime is known to be higher in some European regions than others. However, cross-national surveys, which are the main source of information to map worry about crime across Europe, are designed to be representative of large areas (countries), and regions often suffer from small and unrepresentative sample sizes. This research produces reliable model-based small area estimates of worry about crime at regional level from European Social Survey data, in order to map the phenomenon and examine its macro-level predictors. Model-based small area estimation techniques borrow strength across areas to produce reliable estimates of parameters of interest. Estimates of worry about crime are higher in most South and East European regions, in contrast to Northern and Central Europe

The Ecological Impacts of Contaminated Sediment from Abandoned Metal Mines

Contains public sector information licensed under the Open Government Licence v3.0. The OGL requires that users acknowledge the information provider and/or source of the information with an attribution statement.Pollution from abandoned non-coal (i.e. metal) mines is a serious impediment to rivers meeting the water quality targets set out in River Basin Management Plans. Recent work has identified the mines most likely to be causing a significant environmental impact and hence where efforts to prevent pollution need to be focussed. Yet, it is not clear to what extent rivers, and the animal and plant life they support, are impacted by the legacy of past pollution still bound up in river sediments. Work will be undertaken to reduce toxic metals in mine waters before they enter the river. However, if riverbed sediments are already contaminated and affecting life in rivers, the planned clean-up of mine water sources may not result in recovery of ecological condition. A controlled laboratory experiment was undertaken where river invertebrates (mayfly larvae) from an uncontaminated site were incubated with contaminated riverbed sediment collected downstream of an abandoned metal mine. Concentrations of metals in the tissues of the mayflies increased over the duration of the incubation, particularly those metals that were in high concentrations in the sediment, i.e. cadmium, copper and zinc. As the sediment was the only substantial source of metals in the experiment, it is apparent that the contaminated riverbed sediment was acting as a source of bioavailable metals. It is likely that contaminated sediments, including riverbed sediment, will act as a source of bioavailable metals, at least to benthic organisms, even where mine drainage water is treated to reduce metal concentrations. Metal toxicity occurs when the rate of metal uptake into an organism exceeds the combined rates of excretion and physiological detoxification. Current tests of metal toxicity on biota typically do not match in scale (temporal, spatial and taxonomic range) with assessments of ecological quality undertaken for management, which raises questions regarding the adequacy of environmental limits based on laboratory testing. Existing data were compiled describing geochemistry of riverbed sediment and the Biological Quality Elements invertebrates, diatoms, macrophytes and fish, collected by the regulatory authorities to assess the condition of rivers. As toxic effects of trace metals were not expected at low concentrations, the biological response to sediment metal concentration was determined using a threshold model. Thresholds were found for biotic metrics based on species richness, but other metrics (diatom EQR, macrophyte EQR and invertebrate ASPT) displayed implausible positive relationships with sediment metal concentrations and should not be relied upon for classification of ecological status in waterbodies affected by mining. New data were collected from 20 spatially-independent river catchments in areas affected by metal mine facilities, including samples of the macroinvertebrate community, bioavailability of metals (assessed as metal concentrations in the body tissue of tolerant taxa), and sediment metal concentrations. There were strong correlations between sediment metal concentrations and measured bioavailability, particularly for copper and lead. Measurements of bioavailable metals were related to changes in taxon richness in the invertebrate samples. The data were used to develop a new biotic index (MetTol), which can be used to assess the extent of ecological damage from metal contamination using standard invertebrate monitoring data, and to construct dose response curves based on species sensitivities. A number of approaches were used to establish tolerable limits for sediment metal concentrations based on ecological data, and the results compared with existing Canadian sediment quality guidelines. The limits for copper derived from ecological data were most consistent with existing sediment guidelines. The limits for other metals (silver, arsenic, cadmium, nickel, lead and zinc) derived from ecological data were up to an order of magnitude above the Canadian interim sediment quality guidelines. These existing guidelines, based on toxicological data, may be too precautionary, and we suggest that guideline sediment concentrations based on ecological data may provide a more appropriate level of protection for the environment

Towards the clinical implementation of pharmacogenetics in bipolar disorder.

BackgroundBipolar disorder (BD) is a psychiatric illness defined by pathological alterations between the mood states of mania and depression, causing disability, imposing healthcare costs and elevating the risk of suicide. Although effective treatments for BD exist, variability in outcomes leads to a large number of treatment failures, typically followed by a trial and error process of medication switches that can take years. Pharmacogenetic testing (PGT), by tailoring drug choice to an individual, may personalize and expedite treatment so as to identify more rapidly medications well suited to individual BD patients.DiscussionA number of associations have been made in BD between medication response phenotypes and specific genetic markers. However, to date clinical adoption of PGT has been limited, often citing questions that must be answered before it can be widely utilized. These include: What are the requirements of supporting evidence? How large is a clinically relevant effect? What degree of specificity and sensitivity are required? Does a given marker influence decision making and have clinical utility? In many cases, the answers to these questions remain unknown, and ultimately, the question of whether PGT is valid and useful must be determined empirically. Towards this aim, we have reviewed the literature and selected drug-genotype associations with the strongest evidence for utility in BD.SummaryBased upon these findings, we propose a preliminary panel for use in PGT, and a method by which the results of a PGT panel can be integrated for clinical interpretation. Finally, we argue that based on the sufficiency of accumulated evidence, PGT implementation studies are now warranted. We propose and discuss the design for a randomized clinical trial to test the use of PGT in the treatment of BD

Kaposi's Sarcoma-Associated Herpesvirus-Encoded LANA Down-Regulates IL-22R1 Expression through a Cis-Acting Element within the Promoter Region

Kaposi's sarcoma-associated herpesvirus (KSHV) is considered to be a necessary, but not sufficient, causal agent of Kaposi's sarcoma (KS). All forms of KS are characterized by the proliferation of spindle-shaped cells, and most (>90%) spindle cells from KS lesions are latently infected with KSHV. During KSHV latency, only a few viral genes are expressed. Among those latent genes, the ORF 73 gene encodes the latency-associated nuclear antigen (LANA), which is critical for the establishment and maintenance of the latent KSHV infection. Much evidence suggests that many cytokines can increase the frequency and aggressiveness of KS. In this study, a microarray analysis of KS and normal tissues revealed that multiple cytokines and cytokine receptors are regulated by KSHV latent infection. Of special interest, IL-22R1 transcript level was found to be down-regulated in the KS tissue. To study the possible regulation of IL-22R1 by LANA, the IL-22R1 promoter was constructed and found to contain a LANA-binding site (LBS). LANA was demonstrated to down-regulate IL-22R1 expression via direct binding to the LBS located within the IL-22R1 promoter region. Furthermore, KSHV latently infected cells showed an impaired response to IL-22 stimulation. These results suggest that LANA can regulate host factor expression by directly binding to a cis-acting element within the factor's promoter to benefit latent viral infection and suppression of the antiviral immune response

Progression of duodenal adenomatosis in familial adenomatous polyposis: due to ageing of subjects and advances in technology

Familial adenomatous polyposis patients are at risk of duodenal cancer. Surveillance is indicated and the extent of duodenal polyposis is quantified by the Spigelman staging system. We noticed an impressive increase in high Spigelman stages over the years and therefore decided to investigate whether this increase might be due to the time-lapse since the inception of surveillance or related to improvements in endoscopic imaging and/or changes in dysplasia-reporting. Patients who were investigated by the same endoscopist since 1980 in at least 2 different episodes of technical improvements were eligible. The period 1980–2009 was divided into 4 episodes using the following landmarks: replacement of fibre-endoscopes by video-endoscopes in 1987, change in processors in 1995, change in image resolution in 2000, and change in dysplasia-reporting in 2006. An increase in Spigelman stages from low stages (0–II 100%) to high stages (III 28.1%, IV 43.8%) was seen (median follow-up: 19.5 years). In patients who progressed, a median of 4 years elapsed before progression by one stage occurred and 7 years to progress by two stages. In a mixed-model analysis, both time-lapse and technical improvements were determinant factors for duodenal disease progression. When both factors were introduced in the model, the time-lapse as well as the change in image resolution and dysplasia-ranking contributed consistently in increasing Spigelman scores and stages. The impressive increase in severity of duodenal polyposis is determined by time-lapse, technological advances and change in dysplasia-reporting. These results might call for a revised Spigelman classification

The aged niche disrupts muscle stem cell quiescence

SUMMARY The niche is a conserved regulator of stem cell quiescence and function. During aging, stem cell function declines. To what extent and by which means age-related changes within the niche contribute to this phenomenon are unknown. We demonstrate that the aged muscle stem cell niche, the muscle fiber, expresses FGF2 under homeostatic conditions, driving a subset of satellite cells to break quiescence and lose self-renewing capacity. We show that relatively dormant aged satellite cells robustly express Sprouty1 (spry1), an inhibitor of FGF signalling. Increasing FGF signalling in aged satellite cells under homeostatic conditions by removing spry1, results in the loss of quiescence, satellite cell depletion and diminished regenerative capacity. Conversely, reducing niche-derived FGF activity through inhibition of FGFR1 signalling or overexpression of spry1 in satellite cells prevents their depletion. These experiments identify an age-dependent change in the stem cell niche that directly influences stem cell quiescence and function

Of Asian Forests and European Fields: Eastern U.S. Plant Invasions in a Global Floristic Context

Background: Biogeographic patterns of species invasions hold important clues to solving the recalcitrant ‘who’, ‘where’, and ‘why ’ questions of invasion biology, but the few existing studies make no attempt to distinguish alien floras (all non-native occurrences) from invasive floras (rapidly spreading species of significant management concern), nor have invasion biologists asked whether particular habitats are consistently invaded by species from particular regions. Methodology/Principal Findings: Here I describe the native floristic provenances of the 2629 alien plant taxa of the Eastern Deciduous Forest of the Eastern U.S. (EUS), and contrast these to the subset of 449 taxa that EUS management agencies have labeled ‘invasive’. Although EUS alien plants come from all global floristic regions, nearly half (45%) have native ranges that include central and northern Europe or the Mediterranean (39%). In contrast, EUS invasive species are most likely to come from East Asia (29%), a pattern that is magnified when the invasive pool is restricted to species that are native to a single floristic region (25 % from East Asia, compared to only 11 % from northern/central Europe and 2 % from the Mediterranean). Moreover, East Asian invaders are mostly woody (56%, compared to just 23 % of the total alien flora) and are significantly more likely to invade intact forests and riparian areas than European species, which dominate managed or disturbed ecosystems. Conclusions/Significance: These patterns suggest that the often-invoked ‘imperialist dogma ’ view of global invasion