Giardia Cyst Wall Protein 1 Is a Lectin That Binds to Curled Fibrils of the GalNAc Homopolymer

The infectious and diagnostic stage of Giardia lamblia (also known as G. intestinalis or G. duodenalis) is the cyst. The Giardia cyst wall contains fibrils of a unique β-1,3-linked N-acetylgalactosamine (GalNAc) homopolymer and at least three cyst wall proteins (CWPs) composed of Leu-rich repeats (CWPLRR) and a C-terminal conserved Cys-rich region (CWPCRR). Our goals were to dissect the structure of the cyst wall and determine how it is disrupted during excystation. The intact Giardia cyst wall is thin (~400 nm), easily fractured by sonication, and impermeable to small molecules. Curled fibrils of the GalNAc homopolymer are restricted to a narrow plane and are coated with linear arrays of oval-shaped protein complex. In contrast, cyst walls of Giardia treated with hot alkali to deproteinate fibrils of the GalNAc homopolymer are thick (~1.2 µm), resistant to sonication, and permeable. The deproteinated GalNAc homopolymer, which forms a loose lattice of curled fibrils, is bound by native CWP1 and CWP2, as well as by maltose-binding protein (MBP)-fusions containing the full-length CWP1 or CWP1LRR. In contrast, neither MBP alone nor MBP fused to CWP1CRR bind to the GalNAc homopolymer. Recombinant CWP1 binds to the GalNAc homopolymer within secretory vesicles of Giardia encysting in vitro. Fibrils of the GalNAc homopolymer are exposed during excystation or by treatment of heat-killed cysts with chymotrypsin, while deproteinated fibrils of the GalNAc homopolymer are degraded by extracts of Giardia cysts but not trophozoites. These results show the Leu-rich repeat domain of CWP1 is a lectin that binds to curled fibrils of the GalNAc homopolymer. During excystation, host and Giardia proteases appear to degrade bound CWPs, exposing fibrils of the GalNAc homopolymer that are digested by a stage-specific glycohydrolase. Author SummaryWhile the walls of plants and fungi contain numerous sugar homopolymers (cellulose, chitin, and β-1,3-glucans) and dozens of proteins, the cyst wall of Giardia is relatively simple. The Giardia wall contains a unique homopolymer of β-1,3-linked N-acetylgalactosamine (GalNAc) and at least three cyst wall proteins (CWPs), each of which is composed of Leu-rich repeats and a C-terminal Cys-rich region. The three major discoveries here are: 1) Fibrils of the GalNAc homopolymer are curled and form a lattice that is compressed into a narrow plane by bound protein in intact cyst walls. 2) Leu-rich repeats of CWP1 form a novel lectin domain that is specific for fibrils of the GalNAc homopolymer, which can be isolated by methods used to deproteinate fungal walls. 3) A cyst-specific glycohydrolase is able to degrade deproteinated fibrils of the GalNAc homopolymer. We incorporate these findings into a new curled fiber and lectin model of the intact Giardia cyst wall and a protease and glycohydrolase model of excystation.National Institutes of Health (AI048082, AI44070, GM31318, RR1088

Pressure relieving support surfaces (PRESSURE) trial : cost effectiveness analysis

Objective To assess tire cost effectiveness of alternating pressure mattresses compared with alternating pressure overlays for the prevention of pressure ulcers in patients admitted to hospital. Design Cost effectiveness analysis carried out alongside the pressure relieving support surfaces (PRESSURE) trial; a multicentre UK based pragmatic randomised controlled trial. Setting 11 hospitals in six UK NHS trusts. Participants Intention to treat population comprising 1971 participants. Main outcome measures Kaplan Meier estimates of restricted mean time to development of pressure ulcers and total costs for treatment in hospital. Results Alternating pressure mattresses were associated with lower overall costs (283.6 pound per patient on average, 95% confidence interval -377.59 pound to. 976.79) pound mainly due to reduced length of stay in hospital, and greater benefits (a delay in time to ulceration of 10.64 days on average, - 24.40 to 3.09). The differences in health benefits and total costs for hospital stay between alternating pressure mattresses and alternating pressure overlays were not statistically significant; however, a cost effectiveness acceptability curve indicated that on average alternating pressure mattresses compared with alternating pressure overlays were associated with air 80% probability of being cost saving. Conclusion Alternating pressure mattresses for the prevention of pressure ulcers are more likely to be cost effective and are more acceptable to patients than alternating pressure overlays

Robustness and epistasis in mutation-selection models

We investigate the fitness advantage associated with the robustness of a phenotype against deleterious mutations using deterministic mutation-selection models of quasispecies type equipped with a mesa shaped fitness landscape. We obtain analytic results for the robustness effect which become exact in the limit of infinite sequence length. Thereby, we are able to clarify a seeming contradiction between recent rigorous work and an earlier heuristic treatment based on a mapping to a Schr\"odinger equation. We exploit the quantum mechanical analogy to calculate a correction term for finite sequence lengths and verify our analytic results by numerical studies. In addition, we investigate the occurrence of an error threshold for a general class of epistatic landscape and show that diminishing epistasis is a necessary but not sufficient condition for error threshold behavior.Comment: 20 pages, 14 figure

Long-term changes in population size, distribution and productivity of skuas (Stercorarius spp.) at Signy Island, South Orkney Islands

In this study, we investigate the numbers, productivity and territory distribution of the two species of skuas (brown Stercorarius lonnbergi and south polar Stercorarius maccormicki) breeding at Signy Island, South Orkneys, and compare the results with trends elsewhere. Comparison with previous counts indicates a biphasic increase in brown skuas at Signy Island; much faster from 1958/1959 to 1982/1983 (3.3 % per annum), than in subsequent years (0.4 % per annum from 1983/1984 to 2013/2014). Relative distribution of territories has changed little over time. The reduced rate of population growth in recent years was broadly coincident with a decrease in numbers of penguins (and therefore potential prey), which may also explain recent reductions in skua numbers at other Antarctic sites. As prey have become limiting, breeding success of brown skuas at Signy Island is now slightly lower than in the 1950s/early 1960s, but timing of breeding does not appear to have changed. Brown skuas at Signy Island may still have enough resources to start breeding, but as the season progresses and availability of resources declines, chick survival is reduced. South polar skuas have declined from ten pairs in 1982/1983 to one pair in 2013/2014, and mixed pairs have increased from one to three pairs. A review of the literature indicated that although population trend data are available for relatively few sites elsewhere in the subantarctic and Antarctic, numbers of brown skuas appear to be generally decreasing or stable, and of south polar skuas to be stable or increasing

A massive nebula around the Luminous Blue Variable star RMC143 revealed by ALMA

The luminous blue variable (LBV) RMC143 is located in the outskirts of the 30~Doradus complex, a region rich with interstellar material and hot luminous stars. We report the $3\sigma$ sub-millimetre detection of its circumstellar nebula with ALMA. The observed morphology in the sub-millimetre is different than previously observed with HST and ATCA in the optical and centimetre wavelength regimes. The spectral energy distribution (SED) of RMC143 suggests that two emission mechanisms contribute to the sub-mm emission: optically thin bremsstrahlung and dust. Both the extinction map and the SED are consistent with a dusty massive nebula with a dust mass of $0.055\pm0.018~M_{\odot}$ (assuming $\kappa_{850}=1.7\rm\,cm^{2}\,g^{-1}$). To date, RMC143 has the most dusty LBV nebula observed in the Magellanic Clouds. We have also re-examined the LBV classification of RMC143 based on VLT/X-shooter spectra obtained in 2015/16 and a review of the publication record. The radiative transfer code CMFGEN is used to derive its fundamental stellar parameters. We find an effective temperature of $\sim 8500$~K, luminosity of log$(L/L_{\odot}) = 5.32$, and a relatively high mass-loss rate of $1.0 \times 10^{-5}~M_{\odot}$~yr$^{-1}$. The luminosity is much lower than previously thought, which implies that the current stellar mass of $\sim8~M_{\odot}$ is comparable to its nebular mass of $\sim 5.5~M_{\odot}$ (from an assumed gas-to-dust ratio of 100), suggesting that the star has lost a large fraction of its initial mass in past LBV eruptions or binary interactions. While the star may have been hotter in the past, it is currently not hot enough to ionize its circumstellar nebula. We propose that the nebula is ionized externally by the hot stars in the 30~Doradus star-forming region.Comment: Paper accepted by A&A on 09/05/2019 and in proof stage. Second comments by referee are included in this versio

MutY-Homolog (MYH) inhibition reduces pancreatic cancer cell growth and increases chemosensitivity

Patients with pancreatic ductal adenocarcinoma (PC) have a poor prognosis due to metastases and chemoresistance. PC is characterized by extensive fibrosis, which creates a hypoxic microenvironment, and leads to increased chemoresistance and intracellular oxidative stress. Thus, proteins that protect against oxidative stress are potential therapeutic targets for PC. A key protein that maintains genomic integrity against oxidative damage is MutY-Homolog (MYH). No prior studies have investigated the function of MYH in PC cells. Using siRNA, we showed that knockdown of MYH in PC cells 1) reduced PC cell proliferation and increased apoptosis; 2) further decreased PC cell growth in the presence of oxidative stress and chemotherapy agents (gemcitabine, paclitaxel and vincristine); 3) reduced PC cell metastatic potential; and 4) decreased PC tumor growth in a subcutaneous mouse model in vivo. The results from this study suggest MYH may be a novel therapeutic target for PC that could potentially improve patient outcome by reducing PC cell survival, increasing the efficacy of existing drugs and reducing metastatic spread