Parasites in the changing world – Ten timely examples from the Nordic-Baltic region

The world is changing, and parasites adapt. The Nordic-Baltic region in northern Europe – including the Nordic countries Denmark, Finland, Iceland, Norway and Sweden, and the Baltic States Estonia, Latvia and Lithuania – is facing new parasitological challenges due to changes in populations of parasites and their hosts and the spread of new parasites to the region due to climate change. Some changes can also be ascribed to increased awareness and detection. In this paper, we review and discuss a convenience selection of ten timely examples of recent observations that exemplify trends and challenges from different fields of parasitology, with particular focus on climate change and potential changes in epidemiology of pathogens in northern Europe. The examples illustrate how addressing parasitological challenges often requires both intersectoral and international collaboration, and how using both historical baseline data and modern methodologies are needed.publishedVersio

Microaspiration for high-pressure freezing: a new method for ultrastructural preservation of fragile and sparse tissues for TEM and electron tomography

High-pressure freezing is the preferred method to prepare thick biological specimens for ultrastructural studies. However, the advantages obtained by this method often prove unattainable for samples that are difficult to handle during the freezing and substitution protocols. Delicate and sparse samples are difficult to manipulate and maintain intact throughout the sequence of freezing, infiltration, embedding, and final orientation for sectioning and subsequent TEM imaging. An established approach to surmount these difficulties is the use of cellulose microdialysis tubing to transport the sample. With an inner diameter of 200 micrometers, the tubing protects small and fragile samples within the thickness constraints of high-pressure freezing, and the tube ends can be sealed to avoid loss of sample. Importantly, the transparency of the tubing allows optical study of the specimen at different steps in the process. Here, we describe the use of a micromanipulator and microinjection apparatus to handle and position delicate specimens within the tubing. We report two biologically significant examples that benefit from this approach, 3D cultures of mammary epithelial cells and cochlear outer hair cells. We illustrate the potential for correlative light and electron microscopy as well as electron tomography

Perturbation of the yeast mitochondrial lipidome and associated membrane proteins following heterologous expression of Artemia-ANT

Heterologous expression is a landmark technique for studying a protein itself or its effect on the expression host, in which membrane-embedded proteins are a common choice. Yet, the impact of inserting a foreign protein to the lipid environment of host membranes, has never been addressed. Here we demonstrated that heterologous expression of the Artemia franciscana adenine nucleotide translocase (ANT) in yeasts altered lipidomic composition of their inner mitochondrial membranes. Along with this, activities of complex II, IV and ATP synthase, all membrane-embedded components, were significantly decreased while their expression levels remained unaffected. Although the results represent an individual case of expressing a crustacean protein in yeast inner mitochondrial membranes, it cannot be excluded that host lipidome alterations is a more widespread epiphenomenon, potentially biasing heterologous expression experiments. Finally, our results raise the possibility that not only lipids modulate protein function, but also membrane-embedded proteins modulate lipid composition, thus revealing a reciprocal mode of regulation for these two biomolecular entities

Connexin channels and phospholipids: association and modulation

<p>Abstract</p> <p>Background</p> <p>For membrane proteins, lipids provide a structural framework and means to modulate function. Paired connexin hemichannels form the intercellular channels that compose gap junction plaques while unpaired hemichannels have regulated functions in non-junctional plasma membrane. The importance of interactions between connexin channels and phospholipids is poorly understood.</p> <p>Results</p> <p>Endogenous phospholipids most tightly associated with purified connexin26 or connexin32 hemichannels or with junctional plaques in cell membranes, those likely to have structural and/or modulatory effects, were identified by tandem electrospray ionization-mass spectrometry using class-specific interpretative methods. Phospholipids were characterized by headgroup class, charge, glycerol-alkyl chain linkage and by acyl chain length and saturation. The results indicate that specific endogenous phospholipids are uniquely associated with either connexin26 or connexin32 channels, and some phospholipids are associated with both. Functional effects of the major phospholipid classes on connexin channel activity were assessed by molecular permeability of hemichannels reconstituted into liposomes. Changes to phospholipid composition(s) of the liposome membrane altered the activity of connexin channels in a manner reflecting changes to the surface charge/potential of the membrane and, secondarily, to cholesterol content. Together, the data show that connexin26 and connexin32 channels have a preference for tight association with unique anionic phospholipids, and that these, independent of headgroup, have a positive effect on the activity of both connexin26 and connexin32 channels. Additionally, the data suggest that the likely in vivo phospholipid modulators of connexin channel structure-function that are connexin isoform-specific are found in the cytoplasmic leaflet. A modulatory role for phospholipids that promote negative curvature is also inferred.</p> <p>Conclusion</p> <p>This study is the first to identify (endogenous) phospholipids that tightly associate with connexin channels. The finding that specific phospholipids are associated with different connexin isoforms suggests connexin-specific regulatory and/or structural interactions with lipid membranes. The results are interpreted in light of connexin channel function and cell biology, as informed by current knowledge of lipid-protein interactions and membrane biophysics. The intimate involvement of distinct phospholipids with different connexins contributes to channel structure and/or function, as well as plaque integrity, and to modulation of connexin channels by lipophilic agents.</p

Oncoplastic Breast-Conserving Surgery in Iceland: A Population-Based Study.

To access publisher's full text version of this article click on the hyperlink belowIn Iceland, oncoplastic breast-conservation surgery has been performed since 2008. The aim of this population-based study was to assess and compare the efficacy and patient satisfaction of standard breast-conservation surgery with oncoplastic breast-conservation surgery. This is a population-based, retrospective, observational cohort study on all women undergoing breast-conservation surgery in Iceland from the 1 January 2008 to 31 December 2014. A multivariate logistic regression and linear regression were performed to assess differences in outcomes and a patient satisfaction questionnaire was used to assess certain patient-related outcome measures. A total of 750 women underwent breast-conserving surgery, 665 had standard breast-conservation surgery and 85 oncoplastic breast-conservation surgery. Oncoplastic breast-conservation surgery was associated with a significantly larger mean size (2.4 cm vs 1.7 cm, p < 0.001) and weight (181.8 g vs 63.4 g, p < 0.001) of breast specimen excised when compared to standard breast-conservation surgery. After correcting for confounding factors, there was no significant difference in surgical margin involvement (odds ratio = 0.97, confidence interval = 0.44-1.97), frequency of complications (odds ratio = 1.06, confidence interval = 0.46-2.18), frequency of reoperations (odds ratio = 0.98; confidence interval = 0.50-1.81), or time to first adjuvant therapy (-0.23 days for oncoplastic breast-conservation surgery, p = 0.95). Patient satisfaction was high in both groups, although not statistically different (96% in oncoplastic breast-conservation surgery group vs 89% in the standard breast-conservation surgery group, p = 0.84). Our results show that oncoplastic breast-conservation surgery is at least as safe as standard breast-conservation surgery in selected cases and may be preferable in ductal carcinoma in situ.Visindasjoour Landspitalans (Landspitali University Hospital research fund

Use of Moringa oleifera seed extracts to reduce helminth egg numbers and turbidity in irrigation water

Water from wastewater-polluted streams and dug-outs is the most commonly used water source for irrigation in urban farming in Ghana, but helminth parasite eggs in the water represent health risks when used for crop production. Conventional water treatment is expensive, requires advanced technology and often breaks down in less developed countries so low cost interventions are needed. Field and laboratory based trials were carried out in order to investigate the effect of the natural coagulant Moringa oleifera (MO) seed extracts in reducing helminh eggs and turbidity in irrigation water, turbid water, wastewater and tap water. In medium to high turbid water MO extracts were effective in reducing the number of helminth eggs by 94e99.5% to 1e2 eggs per litre and the turbidity to 7e11 NTU which is an 85e96% reduction. MO is readily available in many tropical countries and can be used by farmers to treat high turbid water for irrigation, however, additional improvements of water quality, e.g. by sand filtration, is suggested to meet the guideline value of 1 helminth egg per litre and a turbidity of 2 NTU as recommended by the World Health Organization and the U.S. Environmental Protection Agency for water intended for irrigation. A positive correlation was established between reduction in turbidity and helminth eggs in irrigation water, turbid water and wastewater treated with MO. This indicates that helminth eggs attach to suspended particles and/or flocs facilitated by MO in the water, and that turbidity and helminth eggs are reduced with the settling flocs. However, more experiments with water samples containing naturally occurring helminth eggs are needed to establish whether turbidity can be used as a proxy for helminth eggs

Questionnaire – Supplemental material for Oncoplastic breast-conserving surgery in iceland: A population-based study

<p>Supplemental material, Questionnaire for Oncoplastic breast-conserving surgery in iceland: A population-based study by E. P. Palsdottir, S. H. L. Lund, K. S. A. Asgeirsson in Scandinavian Journal of Surgery</p