Understanding slow compression of frictional granular particles by network analysis.

We consider frictional granular packings exposed to quasi-static compression rates, with a focus on systems above the jamming transition. For frictionless packings, earlier work (S. Luding et al., Soft Matter, 2022, 18(9), 1868-1884) has uncovered that the system evolution/response involves smooth evolution phases, interrupted by fast transitions (events). The general finding is that the force networks' static quantities correlate closely with the pressure, while their evolution resembles the kinetic energy for both frictionless and frictional packings. The former represents reversible (elastic) particle deformations with affine and non-affine components, whereas the latter also involves much stronger, irreversible (plastic) rearrangements of the packings. Events are associated with jumps in the overall kinetic energy as well as dramatic changes in the force networks describing the particle micro-structure. The frictional nature of particle interactions affects both their frequency and the relevant time scale magnitude. For intermediate friction, events are often followed by an unexpected slow-down during which the kinetic energy drops below its average value. We find that these slow-downs are associated with a significant decrease in the non-affine dynamics of the particles, and are strongly influenced by friction. Friction modifies the structure of the networks, both through the typical number of contacts of a particle, and by influencing topological features of the resulting networks. Furthermore, friction modifies the dynamics of the networks, with larger values of friction leading to smaller changes of the more stable networks.

GRASP negatively regulates the secretion of the virulence factor gp63 in Leishmania.

Metalloprotease-gp63 is a virulence factor secreted by Leishmania. However, secretory pathway in Leishmania is not well defined. Here, we cloned and expressed the GRASP homolog from Leishmania. We found that Leishmania expresses one GRASP homolog of 58 kDa protein (LdGRASP) which localizes in LdRab1- and LPG2-positive Golgi compartment in Leishmania. LdGRASP was found to bind with COPII complex, LdARF1, LdRab1 and LdRab11 indicating its role in ER and Golgi transport in Leishmania. To determine the function of LdGRASP, we generated LdGRASP knockout parasites using CRISPR-Cas9. We found fragmentation of Golgi in Ld:GRASPKO parasites. Our results showed enhanced transport of non-GPI-anchored gp63 to the cell surface leading to higher secretion of this form of gp63 in Ld:GRASPKO parasites in comparison to Ld:WT cells. In contrast, we found that transport of GPI-anchored gp63 to the cell surface is blocked in Ld:GRASPKO parasites and thereby inhibits its secretion. The overexpression of dominant-negative mutant of LdRab1 or LdSar1 in Ld:GRASPKO parasites significantly blocked the secretion of non-GPI-anchored gp63. Interestingly, we found that survival of transgenic parasites overexpressing Ld:GRASP-GFP is significantly compromised in macrophages in comparison to Ld:WT and Ld:GRASPKO parasites. These results demonstrated that LdGRASP differentially regulates Ldgp63 secretory pathway in Leishmania.

Leishmania highjack host lipid body for its proliferation in macrophages by overexpressing host Rab18 and TRAPPC9 by downregulating miR-1914-3p expression.

Lipids stored in lipid-bodies (LBs) in host cells are potential sources of fatty acids for pathogens. However, the mechanism of recruitment of LBs from the host cells by pathogens to acquire fatty acids is not known. Here, we have found that Leishmania specifically upregulates the expression of host Rab18 and its GEF, TRAPPC9 by downregulating the expression of miR-1914-3p by reducing the level of Dicer in macrophages via their metalloprotease gp63. Our results also show that miR-1914-3p negatively regulates the expression of Rab18 and its GEF in cells. Subsequently, Leishmania containing parasitophorous vacuoles (Ld-PVs) recruit and retain host Rab18 and TRAPPC9. Leishmania infection also induces LB biogenesis in host cells and recruits LBs on Ld-PVs and acquires FLC12-labeled fatty acids from LBs. Moreover, overexpression of miR-1914-3p in macrophages significantly inhibits the recruitment of LBs and thereby suppresses the multiplication of parasites in macrophages as parasites are unable to acquire fatty acids. These results demonstrate a novel mechanism how Leishmania acquire fatty acids from LBs for their growth in macrophages.

Evolution of force networks during stick-slip motion of an intruder in a granular material: Topological measures extracted from experimental data.

In quasi-two-dimensional experiments with photoelastic particles confined to an annular region, an intruder constrained to move in a circular path halfway between the annular walls experiences stick-slip dynamics. We discuss the response of the granular medium to the driven intruder, focusing on the evolution of the force network during sticking periods. Because the available experimental data do not include precise information about individual contact forces, we use an approach developed in our previous work [Basak et al., J. Eng. Mech. 147, 04021100 (2021)0733-939910.1061/(ASCE)EM.1943-7889.0002003] based on networks constructed from measurements of the integrated strain magnitude on each particle. These networks are analyzed using topological measures based on persistence diagrams, revealing that force networks evolve smoothly but in a nontrivial manner throughout each sticking period, even though the intruder and granular particles are stationary. Characteristic features of persistence diagrams show identifiable slip precursors. In particular, the number of generators describing the structure and complexity of force networks increases consistently before slips. Key features of the dynamics are similar for granular materials composed of disks or pentagons, but some details are consistently different. In particular, we find significantly larger fluctuations of the measures computed based on persistence diagrams and, therefore, of the underlying networks, for systems of pentagonal particles.

Hemoglobin Endocytosis and Intracellular Trafficking: A Novel Way of Heme Acquisition by Leishmania.

Leishmania species are causative agents of human leishmaniasis, affecting 12 million people annually. Drugs available for leishmaniasis are toxic, and no vaccine is available. Thus, the major thrust is to identify new therapeutic targets. Leishmania is an auxotroph for heme and must acquire heme from the host for its survival. Thus, the major focus has been to understand the heme acquisition process by the parasites in the last few decades. It is conceivable that the parasite is possibly obtaining heme from host hemoprotein, as free heme is not available in the host. Current understanding indicates that Leishmania internalizes hemoglobin (Hb) through a specific receptor by a clathrin-mediated endocytic process and targets it to the parasite lysosomes via the Rab5 and Rab7 regulated endocytic pathway, where it is degraded to generate intracellular heme that is used by the parasite. Subsequently, intra-lysosomal heme is initially transported to the cytosol and is finally delivered to the mitochondria via different heme transporters. Studies using different null mutant parasites showed that these receptors and transporters are essential for the survival of the parasite. Thus, the heme acquisition process in Leishmania may be exploited for the development of novel therapeutics.

On intermittency in sheared granular systems.

We consider a system of granular particles, modeled by two dimensional frictional soft elastic disks, that is exposed to externally applied time-dependent shear stress in a planar Couette geometry. We concentrate on the external forcing that produces intermittent dynamics of stick-slip type. In this regime, the top wall remains almost at rest until the applied stress becomes sufficiently large, and then it slips. We focus on the evolution of the system as it approaches a slip event. Our main finding is that there are two distinct groups of measures describing system behavior before a slip event. The first group consists of global measures defined as system-wide averages at a fixed time. Typical examples of measures in this group are averages of the normal or tangent forces acting between the particles, system size and number of contacts between the particles. These measures do not seem to be sensitive to an approaching slip event. On average, they tend to increase linearly with the force pulling the spring. The second group consists of the time-dependent measures that quantify the evolution of the system on a micro (particle) or mesoscale. Measures in this group first quantify the temporal differences between two states and only then aggregate them to a single number. For example, Wasserstein distance quantitatively measures the changes of the force network as it evolves in time while the number of broken contacts quantifies the evolution of the contact network. The behavior of the measures in the second group changes dramatically before a slip event starts. They increase rapidly as a slip event approaches, indicating a significant increase in fluctuations of the system before a slip event is triggered.

Alternative splicing of ceramide synthase 2 alters levels of specific ceramides and modulates cancer cell proliferation and migration in Luminal B breast cancer subtype.

Global dysregulation of RNA splicing and imbalanced sphingolipid metabolism has emerged as promoters of cancer cell transformation. Here, we present specific signature of alternative splicing (AS) events of sphingolipid genes for each breast cancer subtype from the TCGA-BRCA dataset. We show that ceramide synthase 2 (CERS2) undergoes a unique cassette exon event specifically in Luminal B subtype tumors. We validated this exon 8 skipping event in Luminal B cancer cells compared to normal epithelial cells, and in patient-derived tumor tissues compared to matched normal tissues. Differential AS-based survival analysis shows that this AS event of CERS2 is a poor prognostic factor for Luminal B patients. As Exon 8 corresponds to catalytic Lag1p domain, overexpression of AS transcript of CERS2 in Luminal B cancer cells leads to a reduction in the level of very-long-chain ceramides compared to overexpression of protein-coding (PC) transcript of CERS2. We further demonstrate that this AS event-mediated decrease of very-long-chain ceramides leads to enhanced cancer cell proliferation and migration. Therefore, our results show subtype-specific AS of sphingolipid genes as a regulatory mechanism that deregulates sphingolipids like ceramides in breast tumors, and can be explored further as a suitable therapeutic target.