Gender Differences in Authorship of Clinical Problem-Solving Articles.

Authors of clinical reasoning exercises analyze diagnostic dilemmas and serve as role models of clinical excellence. We investigated the percentage of women authors in the clinical problem-solving series of three general medicine journals from the inaugural article in each series until July 2019. Women were underrepresented among first, last, and all authors. While the percentage of women among first and all authors has increased, women still constituted <40% of all authors and ≤25% of last authors, and there have been no significant increases in women last authors in any of the three journals. Including more women in clinical reasoning exercises is an opportunity to amplify the voices of women as master clinicians.

Identification and Optimization of Novel Small c-Abl Kinase Activators Using Fragment and HTS Methodologies.

Abelson kinase (c-Abl) is a ubiquitously expressed, nonreceptor tyrosine kinase which plays a key role in cell differentiation and survival. It was hypothesized that transient activation of c-Abl kinase via displacement of the N-terminal autoinhibitory "myristoyl latch", may lead to an increased hematopoietic stem cell differentiation. This would increase the numbers of circulating neutrophils and so be an effective treatment for chemotherapy-induced neutropenia. This paper describes the discovery and optimization of a thiazole series of novel small molecule c-Abl activators, initially identified by a high throughput screening. Subsequently, a scaffold-hop, which exploited the improved physicochemical properties of a dihydropyrazole analogue, identified through fragment screening, delivered potent, soluble, cell-active c-Abl activators, which demonstrated the intracellular activation of c-Abl in vivo.

Structurally Diverse Mitochondrial Branched Chain Aminotransferase (BCATm) Leads with Varying Binding Modes Identified by Fragment Screening.

Inhibitors of mitochondrial branched chain aminotransferase (BCATm), identified using fragment screening, are described. This was carried out using a combination of STD-NMR, thermal melt (Tm), and biochemical assays to identify compounds that bound to BCATm, which were subsequently progressed to X-ray crystallography, where a number of exemplars showed significant diversity in their binding modes. The hits identified were supplemented by searching and screening of additional analogues, which enabled the gathering of further X-ray data where the original hits had not produced liganded structures. The fragment hits were optimized using structure-based design, with some transfer of information between series, which enabled the identification of ligand efficient lead molecules with micromolar levels of inhibition, cellular activity, and good solubility.

The Discovery of in Vivo Active Mitochondrial Branched-Chain Aminotransferase (BCATm) Inhibitors by Hybridizing Fragment and HTS Hits.

The hybridization of hits, identified by complementary fragment and high throughput screens, enabled the discovery of the first series of potent inhibitors of mitochondrial branched-chain aminotransferase (BCATm) based on a 2-benzylamino-pyrazolo[1,5-a]pyrimidinone-3-carbonitrile template. Structure-guided growth enabled rapid optimization of potency with maintenance of ligand efficiency, while the focus on physicochemical properties delivered compounds with excellent pharmacokinetic exposure that enabled a proof of concept experiment in mice. Oral administration of 2-((4-chloro-2,6-difluorobenzyl)amino)-7-oxo-5-propyl-4,7-dihydropyrazolo[1,5-a]pyrimidine-3-carbonitrile 61 significantly raised the circulating levels of the branched-chain amino acids leucine, isoleucine, and valine in this acute study.

Practical purification of hydrophilic fragments and lead/drug-like molecules by reverse phase flash chromatography: tips, tricks and contemporary developments.

Practical purifications of hydrophilic fragments and lead/drug-like molecules are reviewed, focussing particularly on the rational use of C-18 reverse phase flash chromatography by exploiting the relationship between retention time and distribution coefficients. Logical process changes have been implemented to suit the particular physical properties of individual molecules, notably, various combinations of dry loading, methanol-buffer gradients, pH control and columns designed for 100% aqueous elution. These have been used to great effect; facilitating effective, predictable and scalable purifications of fragments and lead/drug-like molecules, for which other techniques sometimes fail.

Clinical management where medicine meets management. Down to earth.

Appreciative inquiry is a method that can help NHS staff during periods of change management. Staff turnover and sickness absence are already down at one heart centre that has used the technique. People who have attended the away-days say approachability to senior staff is improved.

The role of eukaryotic initiation factor 2alpha during the metabolic depression associated with estivation.

We have investigated the role of eukaryotic initiation factor 2alpha (eIF2alpha) in two estivating organisms previously shown to downregulate protein synthesis during metabolic depression, the land snail Helix aspersa Müller and the desert frog Neobatrachus sutor Main 1957. We have developed a method using a single antibody (which binds specifically to the phosphorylated, conserved phosphorylation region) by which the total levels of eIF2alpha and the ratio of phosphorylated eIF2alpha [eIF2alpha(P)] to total (phosphorylated and unphosphorylated) eIF2alpha can be determined. In H. aspersa, we have shown that the level of eIF2alpha mRNA expression is unchanged between the awake and estivating states. The amount of total eIF2alpha is the same in the estivating and awake states, and eIF2alpha(P) is undetectable and must represent < or =10% of total eIF2alpha in both states. Conversely, in N. sutor during estivation, the level of total eIF2alpha increases approximately 1.6-fold and the ratio of eIF2alpha(P)/eIF2alpha increases from 0.22+/-0.11 to 0.52+/-0.08, implicating eIF2alpha phosphorylation in the downregulation of protein synthesis during estivation in this animal. The differences in the amounts of eIF2alpha and the level of its phosphorylation between these two species also suggest possible differences either in the mechanism by which protein synthesis is downregulated during estivation or in the sensitivity of the initiation of translation to eIF2alpha(P) levels.

In vivo downregulation of protein synthesis in the snail Helix apersa during estivation.

Protein synthesis is downregulated during metabolic depression in a number of systems where the metabolic depression is effected by obvious extrinsic cues. The metabolic depression of the estivating land snail Helix apersa occurs in the absence of any obvious physiological stress and has an intrinsic component independent of temperature, pH, O(2) status, or osmolality. We show that this metabolic depression is accompanied by a downregulation of protein synthesis in vivo. The rate of protein synthesis decreases in two major tissues during estivation: to 23% and 53% of the awake rate in hepatopancreas and foot muscle, respectively. We show from calculations of the theoretical contribution of protein synthesis to total O(2) consumption that the depression of protein synthesis must be a significant, obligate, in vivo component of metabolic depression in H. aspersa.