Levels of soluble complement regulators predict severity of COVID-19 symptoms.

The SARS-CoV-2 virus continues to cause significant morbidity and mortality worldwide from COVID-19. One of the major challenges of patient management is the broad range of symptoms observed. While the majority of individuals experience relatively mild disease, a significant minority of patients require hospitalisation, with COVID-19 still proving fatal for some. As such, there remains a desperate need to better understand what drives this severe disease, both in terms of the underlying biology, but also to potentially predict at diagnosis which patients are likely to require further interventions, thus enabling better outcomes for both patients and healthcare systems. Several lines of evidence have pointed to dysregulation of the complement cascade as a major factor in severe COVID-19 outcomes. How this is underpinned mechanistically is not known. Here, we have focussed on the role of the soluble complement regulators Complement Factor H (FH), its splice variant Factor H-like 1 (FHL-1) and five Factor H-Related proteins (FHR1-5). Using a targeted mass spectrometry approach, we quantified these proteins in a cohort of 188 plasma samples from controls and SARS-CoV-2 patients taken at diagnosis. This analysis revealed significant elevations in all FHR proteins, but not FH, in patients with more severe disease, particularly FHR2 and FHR5 (FHR2: 1.97-fold, p<0.0001; FHR5: 2.4-fold, p<0.0001). Furthermore, for a subset of 77 SARS-CoV-2 +ve patients we also analysed time course samples taken approximately 28 days post-diagnosis. Here, we see complement regulator levels drop in all individuals with asymptomatic or mild disease, but regulators remain high in those with more severe outcomes, with elevations in FHR2 over baseline levels in this group. These data support the hypothesis that elevation of circulating levels of the FHR family of proteins could predict disease severity in COVID-19 patients, and that the duration of elevation (or lack of immune activation resolution) may be partly responsible for driving poor outcomes in COVID-19.

Global Proteomic Profiling of Embryonic Stem Cells Using iTRAQ Isobaric Tags with LC-MS/MS Quantification.

In this methods chapter, we describe the use of isobaric tags for relative and absolute quantification (iTRAQ) for the differential expression analysis of global proteins between embryonic stem cell samples. This protocol describes how proteins are collected from cell culture, digested and prepared so that peptides are labeled with these isobaric tags. Labeled digests are pooled, fractionated offline, and quantified using liquid chromatography-mass spectrometry (LC-MS). This offline fractionation allows for a greater separation and thus increased identification/quantification of peptides. This combined method enables large-scale, deep penetration into the proteome of embryonic stem cells. During quantification, the relative intensities of label-derived reporter ions represent the relative amount of peptide in each sample. Using search algorithms that integrate the generated data for the identified and quantified peptides allows the relative quantification of proteins in the samples. The isobaric tags can be used in a 4 or 8 multiplexed manner; however, using an 8-plex experimental setup allows for the simultaneous analysis of biological and technical replicates within the same mass spectrometry run, thus minimizing experimental variation and increasing the confidence in any identified expression differences.

Food-related advertisements and food intake among adult men and women.

Television viewing may contribute to obesity via promotion of sedentary behavior and exposure to food-related commercials. However, the mechanisms by which food-related commercials promote food intake are not well understood. Therefore, the purpose of the present study was to examine the impact of television advertisements on food intake according to sex and transportability, or the tendency to become engrossed in what one is viewing. Eighty-three undergraduate students, free of disordered eating symptoms, were stratified by sex and randomly assigned to one of three conditions (food-related advertisements, neutral advertisements, or no advertisements). They were then identified as high or low in transportability according to a median split. A significant interaction was found between advertisement condition and transportability such that those high in transportability ate more in the food than other advertisement conditions. A second interaction was found between sex and transportability with women high in transportability eating more food than women low in transportability irrespective of advertisement condition. No significant main effects of advertisement condition, sex, or transportability were found. Results suggest the importance of studying the impact of individual difference variables on the relationship between food-related advertising and food intake.

The effects of social support and coping on the relationship between social anxiety and eating disorders.

The current study examined the hypotheses that social support and coping moderate and or mediate the relationship between a broad and a narrow form of social anxiety and eating disorder symptoms. One hundred sixty-nine female undergraduates at a private Midwestern university, completed measures of social support, coping, social anxiety, fear of negative evaluation, and disordered eating attitudes and behaviors. Results of hierarchical multiple regression analyses indicated that higher levels of social support are associated with a weaker association between social anxiety and eating disorder symptomatology. Low use of task- and avoidant-oriented (distraction) coping and increased use of emotion-oriented coping are associated with a stronger association between social anxiety and eating disorder symptomatology. Implications for research and clinical intervention are discussed.

Segregating YKU80 and TLC1 alleles underlying natural variation in telomere properties in wild yeast.

In yeast, as in humans, telomere length varies among individuals and is controlled by multiple loci. In a quest to define the extent of variation in telomere length, we screened 112 wild-type Saccharomyces sensu stricto isolates. We found extensive telomere length variation in S. paradoxus isolates. This phenotype correlated with their geographic origin: European strains were observed to have extremely short telomeres (<150 bp), whereas American isolates had telomeres approximately three times as long (>400 bp). Insertions of a URA3 gene near telomeres allowed accurate analysis of individual telomere lengths and telomere position effect (TPE). Crossing the American and European strains resulted in F1 spores with a continuum of telomere lengths consistent with what would be predicted if many quantitative trait loci (QTLs) were involved in length maintenance. Variation in TPE is similarly quantitative but only weakly correlated with telomere length. Genotyping F1 segregants indicated several QTLs associated with telomere length and silencing variation. These QTLs include likely candidate genes but also map to regions where there are no known genes involved in telomeric properties. We detected transgressive segregation for both phenotypes. We validated by reciprocal hemizygosity that YKU80 and TLC1 are telomere-length QTLs in the two S. paradoxus subpopulations. Furthermore, we propose that sequence divergence within the Ku heterodimer generates negative epistasis within one of the allelic combinations (American-YKU70 and European-YKU80) resulting in very short telomeres.

Interpersonal sensitivity predicts bulimic symptomatology cross-sectionally and longitudinally.

Individuals who experience interpersonal sensitivity (IPS) may be at an increased risk for developing eating disorder symptomatology. The purpose of the present study was to assess the predictive capacity of IPS and related constructs in the development of bulimic symptomatology both cross-sectionally and longitudinally while controlling for depressive symptoms. Participants were 119 female undergraduate psychology students attending a private mid-size Midwestern university. Data were collected at both the beginning and end of the academic semester. Participants completed the Center for Epidemiological Study - Depression Scale, Brief Fear of Negative Evaluation Scale, Interpersonal Sensitivity Measure, and the Bulimia Test - Revised. The Fragile Inner Self subscale of the IPSM was found to significantly account for additional variability in BULIT-R scores after controlling for depression both cross-sectionally and longitudinally (8% and 2%, respectively). IPS is a suitable candidate for inclusion in the dual pathway model of bulimic symptomatology.