An investigation into the function of the SUMOylation of Nse2 and PCNA in S. pombe

Small ubiquitin like modifier (SUMO) is post-translationally attached to target proteins, forming a covalent bond between its C-terminal glycine and one or more lysine residues on the target protein. SUMO modification of target proteins can affect protein-protein interactions, protein activity, localistation and stability. This study set out to develop an efficient in vitro SUMOylation system to enable the identification of target lysine residues in S. pombe proteins by mass spectrometry. This involved incorporating a trypsin cleavage site adjacent to the SUMO di-glycine motif to improve peptide coverage during mass spectrometry. Several SUMOylated target proteins were identified here, including the E2 SUMO conjugating enzyme Hus5, the E3 SUMO ligase Nse2 and PCNA. The second part of this study focused on the characterisation of unSUMOylatable E3 SUMO ligase nse2 mutants. Integration of lysine to arginine mutations into the genome did not result in any mutant phenotypes and a function for auto-SUMOylation of Nse2 was not identified. During this study, human patients with mutations in the nse2 gene were reported and the equivalent mutations were integrated into the S. pombe nse2 gene to investigate the effect of the mutations. The final part of this work involved the analysis of the SUMOylation of S. pombe PCNA. Using the in vitro system, four target lysine residues for SUMO were identified. SUMOylation of PCNA was also observed in vivo following pull-down studies and 2D gel analysis of wild type and unSUMOylatable mutants. Extensive epistasis analysis was undertaken using these mutants to investigate the role of SUMOylation of S. pombe PCNA

Stabilizing the Retromer Complex in a Human Stem Cell Model of Alzheimer's Disease Reduces TAU Phosphorylation Independently of Amyloid Precursor Protein.

Developing effective therapeutics for complex diseases such as late-onset, sporadic Alzheimer's disease (SAD) is difficult due to genetic and environmental heterogeneity in the human population and the limitations of existing animal models. Here, we used hiPSC-derived neurons to test a compound that stabilizes the retromer, a highly conserved multiprotein assembly that plays a pivotal role in trafficking molecules through the endosomal network. Using this human-specific system, we have confirmed previous data generated in murine models and show that retromer stabilization has a potentially beneficial effect on amyloid beta generation from human stem cell-derived neurons. We further demonstrate that manipulation of retromer complex levels within neurons affects pathogenic TAU phosphorylation in an amyloid-independent manner. Taken together, our work demonstrates that retromer stabilization is a promising candidate for therapeutic development in AD and highlights the advantages of testing novel compounds in a human-specific, neuronal system

The S. pombe translation initiation factor eIF4G is sumoylated and associates with the SUMO protease Ulp2

SUMO is a small post-translational modifier, that is attached to lysine residues in target proteins. It acts by altering proteinprotein interactions, protein localisation and protein activity. SUMO chains can also act as substrates for ubiquitination, resulting in proteasome-mediated degradation of the target protein. SUMO is removed from target proteins by one of a number of specific proteases. The processes of sumoylation and desumoylation have well documented roles in DNA metabolism and in the maintenance of chromatin structure. To further analyse the role of this modification, we have purified protein complexes containing the S. pombe SUMO protease, Ulp2. These complexes contain proteins required for ribosome biogenesis, RNA stability and protein synthesis. Here we have focussed on two translation initiation factors that we identified as co-purifying with Ulp2, eIF4G and eIF3h. We demonstrate that eIF4G, but not eIF3h, is sumoylated. This modification is increased under conditions that produce cytoplasmic stress granules. Consistent with this we observe partial co-localisation of eIF4G and SUMO in stressed cells. Using HeLa cells, we demonstrate that human eIF4GI is also sumoylated; in vitro studies indicate that human eIF4GI is modified on K1368 and K1588, that are located in the C-terminal eIF4A- and Mnk-binding sites respectively

Evaluation of the impact, treatment patterns, and patient and physician perceptions of vasomotor symptoms associated with menopause in Europe and the United States.

OBJECTIVES This study elicited the views of physicians and patients with vasomotor symptoms (VMS) associated with menopause on the impact of VMS and treatment patterns/perceptions. STUDY DESIGN Data from the Adelphi VMS Disease Specific Programme, a point-in-time survey conducted in 5 European countries and the United States in 2020, were used. Primary care providers (PCPs) and gynecologists seeing ≥3 patients/week with VMS associated with menopause completed a survey and chart review; their patients were invited to complete a survey and questionnaires. MAIN OUTCOME MEASURES Physicians reported treatment patterns and patient-specific symptoms and treatment preferences. Patients described symptoms, impact of VMS, and treatment satisfaction. RESULTS Participants included 115 PCPs and 118 gynecologists. Physicians reviewed the charts of 1816 patients, 854 of whom completed surveys. Moderate/severe impact of VMS on sleep, mood, quality of life, and work/study was reported by 35.8 %, 31.6 %, 23.6 %, and 15.4 % of women, respectively. Based on chart review, 64.8 % of women were currently prescribed treatment for VMS, most commonly hormone therapy (HT; 73.1 %), followed by selective serotonin or serotonin-norepinephrine reuptake inhibitors (31.3 %). Most women (57.3 %) with VMS were eligible for HT but averse to using it. Despite 91.4 % of physicians finding HT to be effective, 62.7 % agreed (slightly-strongly) that their patients are generally reluctant to use it. One-third of women were dissatisfied with VMS control. CONCLUSIONS VMS can considerably impact daily life. Effective treatment options that are better accepted could potentially improve management of VMS and lead to better quality of life for women with VMS associated with menopause. CLINICAL TRIAL REGISTRATION None

Detection of toxins and harmful algal bloom cells in shellfish hatcheries and efforts toward removal

As the start of the supply chain for the aquaculture industry, hatcheries are a crucial component in the success of oyster and northern quahog (hard clam) aquaculture on the East Coast of the US. Intermittent failures in hatchery production slow industry growth and reduce profits. To begin investigations into the possible role of algal toxins in hatchery production failure, post-treatment hatchery water from one research and four commercial hatcheries in lower Chesapeake Bay, USA, was sampled for (1) toxin presence and (2) harmful algal bloom (HAB) cell enumeration. Overall, seven toxin classes, likely produced by six different HAB species, were detected in post- treatment hatchery water, despite a lack of visually identifiable HAB cells within the facility. Toxins detected include pectenotoxin-2, goniodomin A, karlotoxin-1 and karlotoxin-3, okadaic acid and dinophysistoxin-1, azaspiracid-1 and azaspiracid-2, brevetoxin-2, and microcystin-LR. In a second, more targeted study, two batches of source water were followed and sampled at each step of a water-treatment process in the VIMS Aquaculture Genetics and Breeding Technology Center research hatchery in Gloucester Point, Virginia, USA. Two treatment steps showed particular promise for decreasing the concentrations of the three toxins detected in the source water, 24-h circulation through sand filters and activated charcoal filtration. Toxin concentrations of pectenotoxin-2, 3.53 ± 0.56 pg m

Maryse Condé

Bryant, Femi; Person, Jovita; Small, Salima; Urbanovich, Vicky; Wilson, Alithea; Brown, Justin; Curtright, Lauren. (2000). Maryse Condé. Retrieved from the University Digital Conservancy, https://hdl.handle.net/11299/166128

Alcohol, Smoking, and Caffeine in Relation to Fecundability, with Effect Modification by NAT2

Common polymorphisms in the N-acetyltransferase-2 (NAT2) metabolic enzyme determine slow or rapid acetylator phenotypes. We investigated the effects of alcohol, smoking, and caffeine on fecundability, and determined whether the effects were modified by NAT2

Drivers and Challenges in Raising the Achievement of Pupils from Bangladeshi, Somali and Turkish Backgrounds

In this report we examine why attainment rates amongst Bangladeshi pupils have significantly improved over recent years while Turkish and Somali pupils continue to perform below the national average. We explore the experiences of pupils within the education system, the characteristics and policies of their schools, the support received from the local authorities and the impact of parental involvement and parents’ attitudes to school. We also suggest the factors and school strategies likely to have contributed to improved outcomes for underperforming ethnic minority pupils. Evidence will include examples of good practice in schools with high performing pupils from these groups. Wherever possible we identify factors that are specific to the three groups in this study, and in particular the factors that explain increasing attainment rates amongst Bangladeshi pupils