Gadoxetic acid magnetic-enhanced resonance imaging in the diagnosis of cholangiocarcinoma

The use of liver magnetic resonance imaging is increasing thanks to its multiparametric sequences that allow a better tissue characterization, and the use of hepatobiliary contrast agents. This review aims to evaluate gadoxetic acid enhanced magnetic resonance imaging in the diagnosis and staging of cholangiocarcinoma and its different clinical and radiological classifications proposed in the literature. We also analyze the epidemiology, risk factors in correlation with clinical findings and laboratory data.     

Distinct roles of the photosystem II protein PsbS and zeaxanthin in the regulation of light harvesting in plants revealed by fluorescence lifetime snapshots

The photosystem II (PSII) protein PsbS and the enzyme violaxanthin deepoxidase (VDE) are known to influence the dynamics of energy-dependent quenching (qE), the component of nonphotochemical quenching (NPQ) that allows plants to respond to fast fluctuations in light intensity. Although the absence of PsbS and VDE has been shown to change the amount of quenching, there have not been any measurements that can detect whether the presence of these proteins alters the type of quenching that occurs. The chlorophyll fluorescence lifetime probes the excited-state chlorophyll relaxation dynamics and can be used to determine the amount of quenching as well as whether two different genotypes with the same amount of NPQ have similar dynamics of excited-state chlorophyll relaxation. We measured the fluorescence lifetimes on whole leaves of Arabidopsis thaliana throughout the induction and relaxation of NPQ for wild type and the qE mutants, npq4, which lacks PsbS; npq1, which lacks VDE and cannot convert violaxanthin to zeaxanthin; and npq1 npq4, which lacks both VDE and PsbS. These measurements show that although PsbS changes the amount of quenching and the rate at which quenching turns on, it does not affect the relaxation dynamics of excited chlorophyll during quenching. In addition, the data suggest that PsbS responds not only to ΔpH but also to the Δψ across the thylakoid membrane. In contrast, the presence of VDE, which is necessary for the accumulation of zeaxanthin, affects the excited-state chlorophyll relaxation dynamics.Plants regulate light harvesting by photosystem II (PSII) in response to changes in light intensity. One way that plants are able to regulate light harvesting is through turning on and off mechanisms that dissipate excess energy. This energy dissipation is assessed via nonphotochemical quenching (NPQ) measurements of chlorophyll fluorescence. Energy-dependent quenching (qE) is the NPQ process with the fastest kinetics. It turns on and off in seconds to minutes, allowing plants to respond to rapid fluctuations in light intensity, which is thought to reduce photodamage (1, 2).Illumination causes the formation of gradients of electrical potential (Δψ) and of proton concentration (ΔpH) across the thylakoid membrane. Although it has been suggested that Δψ may play a role in qE (3), only ΔpH is thought to trigger different proteins and enzymes to induce qE (4). The major known factors involved in induction of qE are the enzyme violaxanthin deepoxidase (VDE) (5) and the PSII protein PsbS (6). The mutant npq1, which lacks VDE and cannot convert violaxanthin to zeaxanthin, has a phenotype with lower qE compared with the wild type (7). Transient absorption measurements suggest that zeaxanthin may quench excited chlorophyll (8). The npq4 mutant, which lacks PsbS, shows no rapidly reversible quenching of chlorophyll fluorescence, suggesting that PsbS is required for qE in vivo (6). PsbS is pH sensitive (9) but is not thought to bind pigments, and thus is likely not the site of quenching (10). It has therefore been hypothesized that PsbS plays an indirect role in quenching, perhaps facilitating a rearrangement of proteins within the grana (11–13). In this paper, we examine the fluorescence lifetime of chlorophyll throughout the induction and relaxation of quenching in intact leaves with and without PsbS and zeaxanthin to examine whether PsbS and zeaxanthin change the type of quenching that occurs in plants.The amount and dynamics of qE are generally measured by changes in the chlorophyll fluorescence yield. One limitation of the chlorophyll fluorescence yield is that it can only inform on the amount of quenching, not on excited-state chlorophyll relaxation dynamics, which reflect how chlorophyll is quenched. Despite this issue, the amount of quenching is commonly used as a proxy for the type of quenching by separating components of quenching based on kinetics, mutants, and the effects of chemical inhibitors. By artificially increasing ΔpH in isolated chloroplasts from npq4, Johnson and Ruban (14, 15) have been able to increase the amount of quenching in npq4 plants to levels observed in wild type plants, suggesting that PsbS may catalyze qE. One potential complication with these studies is that the use of the chemical mediators of cyclic electron transport often necessitates studying isolated chloroplasts rather than intact leaves. In addition, the observation of equivalent amounts of quenching still does not prove that the type of quenching in npq4 is the same as in wild type.In contrast with fluorescence yield measurements, fluorescence lifetime measurements can be used to determine whether the relaxation dynamics of excited chlorophyll are modified by different mutations, informing on the role of a protein or molecule during quenching. The relaxation dynamics of excited chlorophyll during NPQ depends on many variables, including the distance to a quencher, the interactions between the orbitals of chlorophyll and the quencher, and the number of quenchers (16). The shape of the fluorescence lifetime decay curve can be used to determine whether two samples have similar excited chlorophyll relaxation dynamics. Our results show that, although the presence of PsbS does not alter excited chlorophyll relaxation dynamics, the absence of VDE does. These measurements are performed in intact leaves without any chemical treatments, and the data strongly suggest that PsbS plays a catalytic role in vivo.     

Embryonal tumor with abundant neuropil and true rosettes (ETANTR), ependymoblastoma,and medulloepithelioma share molecular similarity and comprise a single clinicopathological entity

Three histological variants are known within the family of embryonal rosette-forming neuroepithelial brain tumors. These include embryonal tumor with abundant neuropil and true rosettes (ETANTR), ependymoblastoma (EBL), and medulloepithelioma (MEPL). In this study, we performed a comprehensive clinical, pathological, and molecular analysis of 97 cases of these rare brain neoplasms, including genome-wide DNA methylation and copy number profiling of 41 tumors. We identified uniform molecular signatures in all tumors irrespective of histological patterns, indicating that ETANTR, EBL, and MEPL comprise a single biological entity. As such, future WHO classification schemes should consider lumping these variants into a single diagnostic category, such as embryonal tumor with multilayered rosettes (ETMR). We recommend combined LIN28A immunohistochemistry and FISH analysis of the 19q13.42 locus for molecular diagnosis of this tumor category. Recognition of this distinct pediatric brain tumor entity based on the fact that the three histological variants are molecularly and clinically uniform will help to distinguish ETMR from other embryonal CNS tumors and to better understand the biology of these highly aggressive and therapy-resistant pediatric CNS malignancies, possibly leading to alternate treatment strategies.     

p63-microRNA feedback in keratinocyte senescence

We investigated the expression of microRNAs (miRNAs) associated with replicative senescence in human primary keratinocytes. A cohort of miRNAs up-regulated in senescence was identified by genome-wide miRNA profiling, and their change in expression was validated in proliferative versus senescent cells. Among these, miRNA (miR)-138, -181a, -181b, and -130b expression increased with serial passages. miR-138, -181a, and -181b, but not miR-130b, overexpression in proliferating cells was sufficient per se to induce senescence, as evaluated by inhibition of BrdU incorporation and quantification of senescence-activated β-galactosidase staining. We identified Sirt1 as a direct target of miR-138, -181a, and -181b, whereas ΔNp63 expression was inhibited by miR-130b. We also found that ΔNp63α inhibits miR-138, -181a, -181b, and -130b expression by binding directly to p63-responsive elements located in close proximity to the genomic loci of these miRNAs in primary keratinocytes. These findings suggest that changes in miRNA expression, by modulating the levels of regulatory proteins such as p63 and Sirt1, strongly contribute to induction of senescence in primary human keratinocytes, thus linking these two proteins. Our data also indicate that suppression of miR-138, -181a, -181b, and -130b expression is part of a growth-promoting strategy of ΔNp63α in epidermal proliferating cells.