Targeting Mycobacterium tuberculosis CoaBC through Chemical Inhibition of 4'-Phosphopantothenoyl-l-cysteine Synthetase (CoaB) Activity.

Coenzyme A (CoA) is a ubiquitous cofactor present in all living cells and estimated to be required for up to 9% of intracellular enzymatic reactions. Mycobacterium tuberculosis (Mtb) relies on its own ability to biosynthesize CoA to meet the needs of the myriad enzymatic reactions that depend on this cofactor for activity. As such, the pathway to CoA biosynthesis is recognized as a potential source of novel tuberculosis drug targets. In prior work, we genetically validated CoaBC as a bactericidal drug target in Mtb in vitro and in vivo. Here, we describe the identification of compound 1f, a small molecule inhibitor of the 4'-phosphopantothenoyl-l-cysteine synthetase (PPCS; CoaB) domain of the bifunctional Mtb CoaBC, and show that this compound displays on-target activity in Mtb. Compound 1f was found to inhibit CoaBC uncompetitively with respect to 4'-phosphopantothenate, the substrate for the CoaB-catalyzed reaction. Furthermore, metabolomic profiling of wild-type Mtb H37Rv following exposure to compound 1f produced a signature consistent with perturbations in pantothenate and CoA biosynthesis. As the first report of a direct small molecule inhibitor of Mtb CoaBC displaying target-selective whole-cell activity, this study confirms the druggability of CoaBC and chemically validates this target

Inhibiting Mycobacterium tuberculosis CoaBC by targeting an allosteric site.

Coenzyme A (CoA) is a fundamental co-factor for all life, involved in numerous metabolic pathways and cellular processes, and its biosynthetic pathway has raised substantial interest as a drug target against multiple pathogens including Mycobacterium tuberculosis. The biosynthesis of CoA is performed in five steps, with the second and third steps being catalysed in the vast majority of prokaryotes, including M. tuberculosis, by a single bifunctional protein, CoaBC. Depletion of CoaBC was found to be bactericidal in M. tuberculosis. Here we report the first structure of a full-length CoaBC, from the model organism Mycobacterium smegmatis, describe how it is organised as a dodecamer and regulated by CoA thioesters. A high-throughput biochemical screen focusing on CoaB identified two inhibitors with different chemical scaffolds. Hit expansion led to the discovery of potent and selective inhibitors of M. tuberculosis CoaB, which we show to bind to a cryptic allosteric site within CoaB

Autoethnography and severe perineal trauma—an unexpected journey from disembodiment to embodiment

BACKGROUND: There is a lack of research reporting on the physical and emotional experiences of women who sustain severe perineal trauma (third and fourth degree tears). When the researcher identifies with the group being researched, autoethnography can allow an insight into the experiences of the marginalised group through the telling of a personal story. The aim of this paper is to share the journey travelled by an autoethnographer who on examining the issue of severe perineal trauma came to understand the challenges and rewards she experienced through this reflective and analytic process. METHODS: A transformative emancipatory approach guided the design, data collection and analysis of findings from this study. For this paper, a multivocal narrative approach was taken in presenting the findings, which incorporated the words of both the autoethnographer and the twelve women who were interviewed as a component of the study, all of whom had sustained severe perineal trauma. RESULTS: As an autoethnographer, being a member of the group being researched, can be confronting as the necessary reflection upon one’s personal journey may lead to feelings of vulnerability, sadness, and emotional pain. The transformation from disembodied to embodied self, resulted in a physical and emotional breakdown that occurred for this autoethnographer. CONCLUSION: Autoethnographers may experience unexpected emotional and physical challenges as they reflect upon their experiences and research the experiences of others. When incorporating a transformative emancipatory framework, the hardships are somewhat balanced by the rewards of witnessing ‘self-transformation’ as a result of the research

Targeting Mycobacterium tuberculosis CoaBC through Chemical Inhibition of 4'-Phosphopantothenoyl-l-cysteine Synthetase (CoaB) Activity.

Coenzyme A (CoA) is a ubiquitous cofactor present in all living cells and estimated to be required for up to 9% of intracellular enzymatic reactions. Mycobacterium tuberculosis (Mtb) relies on its own ability to biosynthesize CoA to meet the needs of the myriad enzymatic reactions that depend on this cofactor for activity. As such, the pathway to CoA biosynthesis is recognized as a potential source of novel tuberculosis drug targets. In prior work, we genetically validated CoaBC as a bactericidal drug target in Mtb in vitro and in vivo. Here, we describe the identification of compound 1f, a small molecule inhibitor of the 4'-phosphopantothenoyl-l-cysteine synthetase (PPCS; CoaB) domain of the bifunctional Mtb CoaBC, and show that this compound displays on-target activity in Mtb. Compound 1f was found to inhibit CoaBC uncompetitively with respect to 4'-phosphopantothenate, the substrate for the CoaB-catalyzed reaction. Furthermore, metabolomic profiling of wild-type Mtb H37Rv following exposure to compound 1f produced a signature consistent with perturbations in pantothenate and CoA biosynthesis. As the first report of a direct small molecule inhibitor of Mtb CoaBC displaying target-selective whole-cell activity, this study confirms the druggability of CoaBC and chemically validates this target

Cardiovascular and metabolic complications during androgen deprivation: Exercise as a potential countermeasure

Apart from the well-established adverse musculoskeletal and sexual health effects of androgen deprivation therapy (ADT), evidence is accumulating of substantial ADT-related cardiovascular and metabolic complications, which may impact quality of life and overall survival. In this brief review we discuss (1) the incidence of cardiovascular and metabolic complications during/following ADT from large cohort studies, (2) the increased risk factors for cardiovascular and metabolic disease from cross-sectional and prospective studies and (3) the use of physical exercise as a countermeasure in this new era of ADT-related toxicity. It is clear that exercise has the potential to provide a myriad of benefits to men undergoing ADT that may result in reduced morbidity and mortality, and subsequently improve quality of life

Orebody modelling for exploration: the Western Mineralisation, Broken Hill, NSW

The Western Mineralisation in the Broken Hill deposit was studied to identify the zonation sequence of lithogeochemical haloes along and across the strike of the orebody. Samples used are from 77 drill holes and the samples were assayed for Pb, Zn, Fe, S, Cu, Ag, Cd, Sb, Bi and As. Variogram analyses were calculated for all the elements and kriging was used to construct the 3D block model. Analysis of cross sections along and across the strike of the orebody shows that Bi and Sb form broader halos around sulphide masses and this suggests that they are pathfinder elements for the Pb and Zn elements of this orebody. The threshold concentrations (minimum anomaly) of the 10 elements were determined using the concentration-area analysis. On east–west vertical cross sections, the values of linear productivity, variability gradient and zonality index were calculated for each element. Based on the maximum zonality index of each element, the sequence of geochemical zonation pattern was determined from top to bottom of the orebody. The result shows that S, Pb, Zn and Cd tend to concentrate in the upper part of the mineralisation whereas Ag, Cu, Bi and As have a tendency to concentrate in the lower part of the mineralised rocks. Also, an empirical product ratio index was developed based on the position of the elements in the zonation sequence. The methods and results of this research are applicable to exploration of similar Zn and Pb sulphide ore deposits.Mohammad Lotfolah Hamedani, Ian Rutherford Plimer, and Chaoshui X