Status of Women and Gender Equity at CWRU - 2021

The first bi-annual report from the Flora Stone Mather Center reviews the representation of women as well as the intersections of race/ethnicity and gender identities across CWRU executive leadership, faculty, staff, and students.https://commons.case.edu/mathercenter-briefs/1003/thumbnail.jp

WISER: Women in Science and Engineering Roundtable

This brief provides an overview of Case Western Reserve University\u27s Women in Science and Engineering Roundtable, including student evaluation and national metrics about women in STEMhttps://commons.case.edu/mathercenter-briefs/1000/thumbnail.jp

pH-Induced Folding of the Caspase-Cleaved Par-4 Tumor Suppressor: Evidence of Structure Outside of the Coiled Coil Domain

Prostate apoptosis response-4 (Par-4) is a 38 kDa largely intrinsically disordered tumor suppressor protein that functions in cancer cell apoptosis. Par-4 down-regulation is often observed in cancer while up-regulation is characteristic of neurodegenerative conditions such as Alzheimer’s disease. Cleavage of Par-4 by caspase-3 activates tumor suppression via formation of an approximately 25 kDa fragment (cl-Par-4) that enters the nucleus and inhibits Bcl-2 and NF-ƙB, which function in pro-survival pathways. Here, we have investigated the structure of cl-Par-4 using biophysical techniques including circular dichroism (CD) spectroscopy, dynamic light scattering (DLS), and intrinsic tyrosine fluorescence. The results demonstrate pH-dependent folding of cl-Par-4, with high disorder and aggregation at neutral pH, but a largely folded, non-aggregated conformation at acidic p

A Pathway to Solving the Structure of cl-Par-4 Tumor Suppressor Protein: Challenges & Findings

Prostate apoptosis response-4 (Par-4) is a pro-apoptotic tumor suppressor protein. Down-regulation of this protein has been reported in a myriad of cancers. Conversely, up-regulation of Par-4 is found to be associated with several neurodegenerative disorders. Par-4 is unique in the sense it can selectively induce apoptosis in cancer cells. For this, caspase-dependent intracellular cleavage of Par-4 is essential to produce the functionally active fragment, cl-Par-4 (caspase-cleaved Par-4). The cl-Par-4 protein inhibits the NF-κB-mediated cell survival pathway and causes selective apoptosis in various tumor cells. Our laboratory is interested in determining the structure of cl-Par-4 and understanding it’s interaction with various proteins. Currently, we are using biophysical techniques such as circular dichroism (CD) spectroscopy, dynamic light scattering (DLS), and SDS-PAGE to characterize the structure of cl-Par-4 in the presence of various concentrations of monovalent and divalent ions, in order to shed light on the possible ion-specific role of cl-Par-4 in inducing structure and self-association of this protein. Results show that effects on cl-Par-4 conformation are ion-specific, and effects of divalent cations are considerably more pronounced than effects from monovalent cations. We have also found that the cl-Par-4 shows a better stability in presence of Cl- ions than in presence of SO42- ions. Further, with the help of D313K mutant of cl-Par-4, we investigated that charge-charge repulsion between similar charged amino acid residues in leucine zipper is responsible for high salt at neutral pH or low salt at low pH requirement of cl-Par-4. All these findings will be helpful in getting the structured conformation of cl-Par-4 and, therefore, determining the structure of this protein via X-ray crystallography or via nuclear magnetic resonance (NMR).https://digitalcommons.odu.edu/gradposters2022_sciences/1013/thumbnail.jp

Prevalence of malnutrition comparing the GLIM criteria, ESPEN definition and MST malnutrition risk in geriatric rehabilitation patients: RESORT

Background & aims: The Global Leadership Initiative on Malnutrition (GLIM) has developed new criteria for the diagnosis of malnutrition. This study aimed 1) to determine and compare malnutrition prevalence and risk using the GLIM criteria, European Society for Clinical Nutrition and Metabolism (ESPEN) definition of malnutrition and the Malnutrition Screening Tool (MST) in patients admitted to subacute geriatric rehabilitation wards, 2) to explore the agreement of malnutrition prevalence determined by each definition, and 3) to determine the accuracy of the MST against the GLIM criteria and ESPEN definition as references. Methods: Geriatric rehabilitation patients (n = 444) from the observational, longitudinal REStORing health of acutely unwell adulTs (RESORT) cohort in Melbourne, Australia were included. The GLIM criteria, ESPEN definition and MST were applied. Accuracy was determined by the sensitivity, specificity and Area Under the Curve (AUC). Results: According to the GLIM criteria, the overall prevalence of malnutrition was 52.0%. The ESPEN definition diagnosed 12.6% of patients as malnourished and the MST identified 44.4% of patients at risk for malnutrition. Agreement was low; 7% of patients were malnourished and at risk for malnutrition according to all three definitions. The accuracy of the MST compared to the GLIM criteria was fair (sensitivity 56.7%, specificity 69.0%) and sufficient (AUC 0.63); MST compared to the ESPEN definition was fair (sensitivity 60.7%, specificity 58.0%) and poor (AUC 0.59). Conclusions: According to the GLIM criteria, half of geriatric rehabilitation patients were malnourished, whereas the prevalence was much lower applying the ESPEN definition. This highlights the need for further studies to determine diagnostic accuracy of the GLIM criteria compared to pre-existing validated tools

Multi-objective improvement of software using co-evolution and smart seeding

Optimising non-functional properties of software is an important part of the implementation process. One such property is execution time, and compilers target a reduction in execution time using a variety of optimisation techniques. Compiler optimisation is not always able to produce semantically equivalent alternatives that improve execution times, even if such alternatives are known to exist. Often, this is due to the local nature of such optimisations. In this paper we present a novel framework for optimising existing software using a hybrid of evolutionary optimisation techniques. Given as input the implementation of a program or function, we use Genetic Programming to evolve a new semantically equivalent version, optimised to reduce execution time subject to a given probability distribution of inputs. We employ a co-evolved population of test cases to encourage the preservation of the program’s semantics, and exploit the original program through seeding of the population in order to focus the search. We carry out experiments to identify the important factors in maximising efficiency gains. Although in this work we have optimised execution time, other non-functional criteria could be optimised in a similar manner

Ultrafast Excited-State Dynamics of Rhenium(I) Photosensitizers [Re(Cl)(CO)_(3)(N,N)] and [Re(imidazole)(CO)_(3)(N,N)]^+: Diimine Effects

Femto- to picosecond excited-state dynamics of the complexes [Re(L)(CO)_(3)(N,N)]^n (N,N = bpy, phen, 4,7-dimethyl-phen (dmp); L = Cl, n = 0; L = imidazole, n = 1+) were investigated using fluorescence up-conversion, transient absorption in the 650−285 nm range (using broad-band UV probe pulses around 300 nm) and picosecond time-resolved IR (TRIR) spectroscopy in the region of CO stretching vibrations. Optically populated singlet charge-transfer (CT) state(s) undergo femtosecond intersystem crossing to at least two hot triplet states with a rate that is faster in Cl (~100 fs)^(−1) than in imidazole (~150 fs)^(−1) complexes but essentially independent of the N,N ligand. TRIR spectra indicate the presence of two long-lived triplet states that are populated simultaneously and equilibrate in a few picoseconds. The minor state accounts for less than 20% of the relaxed excited population. UV−vis transient spectra were assigned using open-shell time-dependent density functional theory calculations on the lowest triplet CT state. Visible excited-state absorption originates mostly from mixed L;N,N^(•−) → Re^(II) ligand-to-metal CT transitions. Excited bpy complexes show the characteristic sharp near-UV band (Cl, 373 nm; imH, 365 nm) due to two predominantly ππ*(bpy^(•−)) transitions. For phen and dmp, the UV excited-state absorption occurs at 305 nm, originating from a series of mixed ππ* and Re → CO;N,N•− MLCT transitions. UV−vis transient absorption features exhibit small intensity- and band-shape changes occurring with several lifetimes in the 1−5 ps range, while TRIR bands show small intensity changes (≤5 ps) and shifts (~1 and 6−10 ps) to higher wavenumbers. These spectral changes are attributable to convoluted electronic and vibrational relaxation steps and equilibration between the two lowest triplets. Still slower changes (≥15 ps), manifested mostly by the excited-state UV band, probably involve local-solvent restructuring. Implications of the observed excited-state behavior for the development and use of Re-based sensitizers and probes are discussed

Feasibility study of a randomised controlled trial to investigate the treatment of sarcoidosis-associated fatigue with methylphenidate (FaST-MP): a study protocol

Introduction: Fatigue is a frequent and troublesome manifestation of chronic sarcoidosis. This symptom can be debilitating and difficult to treat, with poor response to the treatment. Symptomatic management with neurostimulants, such as methylphenidate, is a possible treatment option. The use of such treatment strategies is not without precedent and has been trialled in cancer-related fatigue. Their use in sarcoidosis requires further evaluation before it can be recommended for clinical practice. Methods and analysis: The Fatigue and Sarcoidosis—Treatment with Methylphenidate study is a randomised, controlled, parallel-arm and feasibility trial of methylphenidate for the treatment of sarcoidosis-associated fatigue. Patients are eligible if they have a diagnosis of sarcoidosis, significant fatigue (measured using the Fatigue Assessment Scale) and have stable disease. Up to 30 participants will be randomly assigned to either methylphenidate (20 mg two times per day) or identical placebo in a 3:2 ratio for 24 weeks. The primary objective is to collect data determining the feasibility of a future study powered to determine the clinical efficacy of methylphenidate for sarcoidosis-associated fatigue. The trial is presently open and will continue until July 2018. Ethics and dissemination: Ethical approval for the study was granted by the Cambridge Central Research Ethics Committee on 21 June 2016 (reference 16/EE/0087) and was approved and sponsored by the Norfolk and Norwich University Hospital (reference 190280). Clinical Trial Authorisation (EudraCT number 2016-000342-60) from the Medicines and Healthcare products Regulatory Agency (MHRA) was granted on 19 April 2016. Results will be presented at relevant conferences and submitted to appropriate journals following trial closure and analysis

Characterization of Cl-Par-4: WT vs. Mutant

Intrinsically disordered proteins (IDPs) play important roles in regulation of cell signaling pathways as well as cellular processes. Dysregulation of these proteins is associated with several human diseases. Prostate apoptosis response-4 (Par-4), a proapoptotic tumor suppressor protein, is categorized as an intrinsically disordered protein and downregulation of this protein has been reported in myriad of cancers including glioma, breast cancers, and prostate cancers. The caspase-cleaved fragment of Par-4 (cl-Par-4) plays an active role in tumor suppression by inhibiting several cell survival pathways. Here, we employed site-directed mutagenesis to introduce a point mutation in the cl-Par-4 wildtype (WT) to generate the D313K cl-Par-4 mutant. We have characterized both the mutant and the WT using various biophysical techniques such as CD, DLS, and NMR to determine the effect of the D313K mutation. The results show that D313K cl-Par-4 attains a compact and well-folded helical conformation, possibly a tetramer, similar to that of the WT in presence of high salt at physiological pH. However, D313K does so with half the amount of salt required for the WT. This establishes that the substitution of a basic residue for an acidic residue at position 313 alleviates inter-helical charge repulsion between dimer partners and helps to stabilize the structural conformation.https://digitalcommons.odu.edu/gradposters2023_gradschool/1004/thumbnail.jp