Sphingosine kinases : emerging structure function insights

Sphingosine kinases (SK1 and SK2) catalyse the conversion of sphingosine into sphingosine 1-phosphate and control fundamental cellular processes, including cell survival, proliferation, differentiation, migration, and immune function. In this review, we highlight recent breakthroughs in the structural and functional characterisation of SK1 and these are contextualised by analysis of crystal structures for closely related prokaryotic lipid kinases. We identify a putative dimerisation interface and propose novel regulatory mechanisms governing structural plasticity induced by phosphorylation and interaction with phospholipids and proteins. Our analysis suggests that the catalytic function and regulation of the enzymes might be dependent on conformational mobility and it provides a roadmap for future interrogation of SK1 function and its role in physiology and disease

Rapid antimicrobial sensitivity testing by single cell nanoscale optical interference

Growing antimicrobial resistance (AMR) is a serious global threat to human health, with estimates of AMR leading to 10 million deaths per year and costing the global economy $100tn by 20501,2. Current methods to detect resistance include phenotypic antibiotic sensitivity testing (AST) which measures bacterial growth and is therefore hampered by slow time to result (~12-24 hours). Therefore new rapid phenotypic methods for AST are urgently needed3. Here we describe a novel method for detecting phenotypic antibiotic resistance in ~45 minutes, capable of detecting single bacteria. The method uses a sensitive laser and detector system to measure nanoscale optical interference of single bacterial cells present in media, with simple sample preparation. This provides a read out of bacterial antibiotic resistance by detecting growth (resistant) or death (sensitive), much faster than current methods. We demonstrate the potential of this technique by determining resistance in both lab and clinical strains of E. coli, a key species for clinically burdensome urinary tract infections. This work provides the basis for a simple and fast diagnostic tool to detect antibiotic resistance in bacteria, reducing the health and economic burdens of AMR

Structure-function analysis of lipid substrates and inhibitors of sphingosine kinases

The sphingosine kinases, SK1 and SK2, catalyse the formation of the bioactive signalling lipid, sphingosine 1-phosphate (S1P), from sphingosine. SK1 and SK2 differ in their subcellular localisation, trafficking and regulation, but the isoforms are also distinct in their selectivity toward naturally occurring and synthetic ligands as substrates and inhibitors. To date, only the structure of SK1 has been determined, and a structural basis for selectivity differences in substrate handling by SK2 has yet to be established. Here we present a structural rationale, based on homology modelling and ligand docking, to account for the capacity of SK2, but not SK1, to efficiently process the pharmacologically active substances, fingolimod (FTY720) and safingol, as substrates. We propose that two key residue differences in hSK2 (Ser305/Thr584 in place of Ala175/Ala339 in hSK1) facilitate conformational switching in the lipid head group anchor residue, Asp308 (corresponding to Asp178 in hSK1), to accommodate substrate diversity for SK2. Our analysis accounts for the contrasting behaviour of fingolimod and safingol as non-turnover inhibitors of SK1, but substrates for SK2, and the observed stereoselectivity for phosphorylation of the pro-S hydroxymethyl group of fingolimod to generate (S)-FTY720-P in vivo. We also rationalise why methylation of the pro-R hydroxymethyl of FTY720 switches the behaviour of the resulting compound, (R)-FTY720 methyl ether (ROMe), to SK2-selective inhibition. Whilst the pharmacological significance of (S)-FTY720-P is firmly established, as the active principle of fingolimod in treating relapsing-remitting multiple sclerosis, the potential importance of SK-mediated phosphorylation of other substrates, such as safingol and non-canonical naturally occuring substrates such as (4E,nZ)-sphingadienes, is less widely appreciated. Thus, the contribution of SK2-derived safingol 1-phosphate to the anti-cancer activity of safingol should be considered. Similarly, the biological role of sphingadiene 1-phosphates derived from plant-based dietary sphingadienes, which we also show here are substrates for both SK1 and SK2, merits investigation

Sphingosine kinases as druggable targets

There is substantial evidence that the enzymes, sphingosine kinase 1 and 2, which catalyse the formation of the bioactive lipid, sphingosine 1-phosphate are involved in physiological and pathophysiological processes. In this chapter, we appraise the evidence that both enzymes are druggable and describe how isoform-specific inhibitors can be developed based on the plasticity of the sphingosine-binding site. This is contextualised with the effect of sphingosine kinase inhibitors in cancer, pulmonary hypertension, neurodegeneration, inflammation and sickling

Second Order Perturbations of Flat Dust FLRW Universes with a Cosmological Constant

We summarize recent results concerning the evolution of second order perturbations in flat dust irrotational FLRW models with $\Lambda\ne 0$. We show that asymptotically these perturbations tend to constants in time, in agreement with the cosmic no-hair conjecture. We solve numerically the second order scalar perturbation equation, and very briefly discuss its all time behaviour and some possible implications for the structure formation.Comment: 6 pages, 1 figure. to be published in "Proceedings of the 5th Alexander Friedmann Seminar on Gravitation and Cosmology", Int. Journ. Mod. Phys. A (2002). Macros: ws-ijmpa.cls, ws-p9-75x6-50.cl

A Mixture of Regressions Model of COVID-19 Death Rates and Population Comorbidities

As the COVID-19 pandemic spread worldwide, it has become clearer that prevalence of certain comorbidities in a given population could make it more vulnerable to serious outcomes of that disease, including fatality. Indeed, it might be insightful from a health policy perspective to identify clusters of populations in terms of the associations between their prevalent comorbidities and the observed COVID-19 specific death rates. In this study, we described a mixture of polynomial time series (MoPTS) model to simultaneously identify (a) three clusters of 86 U.S. cities in terms of their dynamic death rates, and (b) the different associations of those rates with 5 key comorbidities among the populations in the clusters. We also described an EM algorithm for efficient maximum likelihood estimation of the model parameters

Alterations in gut microbiota caused by major depressive disorder or a low FODMAP diet and where they overlap

Beneficial changes in microbiota observed in individuals with a major depressive disorder (MDD) may be initiated with a low fermentable oligosaccharide, disaccharide, monosaccharide, and polyol (FODMAP) elimination diet. Academic Search Ultimate, APA PsychINFO, Cochrane Library, MEDLINE, Scopus and Web of Science were searched for original research documenting differences in microbiota in MDD or changes with a low FODMAP diet in adults (age 18 years +). Studies with fecal microbiota, 16 s RNA sequencing and QIIME pipelines were included. Studies using antibiotics, probiotics, and medications such as antidepressants were excluded. Additionally, studies based on a single gender were excluded as gender impacts microbiota changes in MDD. Four studies addressed differences in microbiota with MDD and another four assessed shifts occurring with a low FODMAP diet. The abundance of Bacteroidetes, Bacteroidaceae and Bacteroides were lower in individuals with MDD but increased with a low FODMAP diet. Abundance of Ruminoccaceae was lower and Bilophila was higher with both a low FODMAP diet and MDD. These results provide preliminary evidence that a low FODMAP diet might drive changes in microbiota that also benefit people with MDD. Further research to assess whether a low FODMAP diet can treat MDD through modification of targeted microbiota is warranted.</p

A Novel Selective Sphingosine Kinase 2 Inhibitor, HWG-35D, Ameliorates the Severity of Imiquimod-Induced Psoriasis Model by Blocking Th17 Differentiation of Naïve CD4 T Lymphocytes

Sphingosine kinases (SK) catalyze the phosphorylation of sphingosine to generate sphingosine-1-phosphate. Two isoforms of SK (SK1 and SK2) exist in mammals. Previously, we showed the beneficial effects of SK2 inhibition, using ABC294640, in a psoriasis mouse model. However, ABC294640 also induces the degradation of SK1 and dihydroceramide desaturase 1 (DES1). Considering these additional effects of ABC294640, we re-examined the efficacy of SK2 inhibition in an IMQ-induced psoriasis mouse model using a novel SK2 inhibitor, HWG-35D, which exhibits nM potency and 100-fold selectivity for SK2 over SK1. Topical application of HWG-35D ameliorated IMQ-induced skin lesions and normalized the serum interleukin-17A levels elevated by IMQ. Application of HWG-35D also decreased skin mRNA levels of interleukin-17A, K6 and K16 genes induced by IMQ. Consistent with the previous data using ABC294640, HWG-35D also blocked T helper type 17 differentiation of naive CD4+ T cells with concomitant reduction of SOCS1. Importantly, HWG-35D did not affect SK1 or DES1 expression levels. These results reaffirm an important role of SK2 in the T helper type 17 response and suggest that highly selective and potent SK2 inhibitors such as HWG-35D might be of therapeutic use for the treatment of psoriasis