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Three-dimensional (3D) ex vivo cultures allow the study of cancer progression and drug resistance mechanisms. Here, we present a protocol for measuring on-target drug sensitivity in a scaffold-free 3D culture system through quantification of apoptotic tumor cells. We provide detailed steps for sample processing, immunofluorescence staining, semi-high-throughput confocal imaging, and imaged-based quantification of 3D cultures. This protocol is versatile and can be applied in principle to any patient-derived material., Competing Interests: Declaration of interests The authors declare no competing interests., (Copyright © 2024 The Author(s). Published by Elsevier Inc. All rights reserved.)

Superconducting circuits reveal themselves as promising physical devices with multiple uses. Within those uses, the fundamental concept of the geometric phase accumulated by the state of a system shows up recurrently, as, for example, in the construction of geometric gates. Given this framework, we study the geometric phases acquired by a paradigmatic setup: a transmon coupled to a superconductor resonating cavity. We do so both for the case in which the evolution is unitary and when it is subjected to dissipative effects. These models offer a comprehensive quantum description of an anharmonic system interacting with a single mode of the electromagnetic field within a perfect or dissipative cavity, respectively. In the dissipative model, the non-unitary effects arise from dephasing, relaxation, and decay of the transmon coupled to its environment. Our approach enables a comparison of the geometric phases obtained in these models, leading to a thorough understanding of the corrections introduced by the presence of the environment.

Shortcuts to adiabaticity (STA) are relevant in the context of quantum systems, particularly regarding their control when they are subjected to time-dependent external conditions. In this paper, we investigate the completion of a nonadiabatic evolution into a shortcut to adiabaticity for a quantum field confined within a one-dimensional cavity containing two movable mirrors. Expanding upon our prior research, we characterize the field's state using two Moore functions that enables us to apply reverse engineering techniques in constructing the STA. Regardless of the initial evolution, we achieve a smooth extension of the Moore functions that implements the STA. This extension facilitates the computation of the mirrors' trajectories based on the aforementioned functions. Additionally, we draw attention to the existence of a comparable problem within nonrelativistic quantum mechanics.

The development of quantum technologies present important challenges such as the need for fast and precise protocols for implementing quantum operations. Shortcuts to adiabaticity (STAs) are a powerful tool for achieving these goals, as they enable us to perform an exactly adiabatic evolution in finite time. In this paper, we present a shortcut to adiabaticity for the control of an optomechanical cavity with two moving mirrors. Given reference trajectories for the mirrors, we find analytical expressions that give us effective trajectories which implement an STA for the quantum field inside the cavity. We then solve these equations numerically for different reference protocols, such as expansions, contractions and rigid motions, thus confirming the successful implementation of the STA and finding some general features of these effective trajectories.

The molecular repertoire promoting cancer cell plasticity is not fully elucidated. Here, we propose that glycosphingolipids (GSLs), specifically the globo and ganglio series, correlate and promote the transition between epithelial and mesenchymal cells. The epithelial character of ovarian cancer remains stable throughout disease progression, and spatial glycosphingolipidomics reveals elevated globosides in the tumor compartment compared with the ganglioside-rich stroma. CRISPR-Cas9 knockin mediated truncation of endogenous E-cadherin induces epithelial-to-mesenchymal transition (EMT) and decreases globosides. The transcriptomics analysis identifies the ganglioside-synthesizing enzyme ST8SIA1 to be consistently elevated in mesenchymal-like samples, predicting poor outcome. Subsequent deletion of ST8SIA1 induces epithelial cell features through mTOR S2448 phosphorylation, whereas loss of globosides in ΔA4GALT cells, resulting in EMT, is accompanied by increased ERK Y202/T204 and AKT S124 . The GSL composition dynamics corroborate cancer cell plasticity, and further evidence suggests that mesenchymal cells are maintained through ganglioside-dependent, calcium-mediated mechanisms., Competing Interests: Declaration of interests The authors declare no competing interests., (Copyright © 2022 The Author(s). Published by Elsevier Inc. All rights reserved.)

Schistosomiasis is a major neglected tropical disease with more than 200 million infections annually. Despite only one drug, praziquantel, being available, the drug pipeline against schistosomiasis is empty, and drug screening tools have limitations. We evaluated the potential of human liver microtissues (hLiMTs) in antischistosomal drug discovery. Because hLiMTs express all human P450 enzymes, they are an excellent tool to evaluate compounds' bioinactivation, bioactivation, and toxicity. To validate the metabolic conversion capacity of hLiMTs, we first quantified ( R )- and ( S )-praziquantel and the main metabolite trans -OH-praziquantel following incubation with 0.032-50 μM (0.01-15.62 μg/mL) praziquantel for up to 72 h by a validated LC-MS/MS method. We cocultured hLiMTs with newly transformed schistosomula (NTS) and evaluated the antischistosomal activity and cytotoxicity of three prodrugs terfenadine, tamoxifen citrate, and flutamide. HLiMTs converted 300-350 ng ( R )-praziquantel within 24 h into trans -OH-praziquantel. We observed changes in the IC 50 values for terfenadine, flutamide, and tamoxifen citrate in comparison to the standard NTS assay in vitro. Cytotoxicity was observed at high concentrations of flutamide and tamoxifen citrate. An in vitro platform containing hLiMTs could serve as an advanced drug screening tool for Schistosoma mansoni , providing information on reduced or increased activity and toxicity.

Schistosomiasis is an acute and chronic disease caused by tropical parasitic worms of the genus Schistosoma, which parasitizes annually over 200 million people worldwide. Screening of antischistosomal compounds is hampered by the low throughput and potential subjectivity of the visual evaluation of the parasite phenotypes, which affects the current drug assays. Here, an impedance-based platform, capable of assessing the viability of Schistosoma mansoni schistosomula exposed to drugs, is presented. This automated and parallelized platform enables unbiased and continuous measurements of dose-response relationships for more than 48 h. The platform performance is established by exposure of schistosomula to three test compounds, praziquantel, oxethazaine, and mefloquine, which are known to affect the larvae phenotypes. The system is thereafter used to investigate the response of schistosomula to methiothepine, an antipsychotic compound, which causes complex drug-induced effects. Continuous monitoring of the parasites reveals transient behavioral phenotypes and allows for extracting temporal characteristics of dose-response curves, which are essential for selecting drugs that feature high activity and fast kinetics of action. These measurements demonstrate that impedance-based detection provides a wealth of information for the in vitro characterization of candidate antischistosomals and, represents a promising tool for the identification of new lead compounds., (© 2020 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

Schistosomiasis is a global disease of significant public health relevance. Only one racemic drug, praziquantel, characterized by low bioavailability, low water solubility and extensive first pass metabolism, is currently available. We studied a new praziquantel formulation (polymorph B), which is based on a racemic praziquantel crystalline polymorph (TELCEU01). Its in vitro activity was tested on newly transformed schistosomula (NTS) and adult Schistosoma mansoni. In vivo studies were conducted in mice harboring chronic S. mansoni infections. Pharmacokinetic (PK) profiles of R- and S-praziquantel and R- and S- polymorph B following oral administration with both formulations were generated by sampling mice at 30, 60, 240 min and 24 h post-treatment, followed by LC-MS/MS analysis. PK parameters were calculated using a non-compartmental analysis with a linear trapezoidal model. In vitro, commercial praziquantel and the polymorph B performed similarly on both NTS (IC 50  = 2.58 and 2.40 µg/mL at 72 h) and adults (IC 50  = 0.05 and 0.07 µg/mL at 72 h). Praziquantel showed higher in vivo efficacy with an ED 50 of 58.75 mg/kg compared to an ED 50 of 122.61 mg/kg for the polymorph B. The PK profiles of the two drugs exhibited differences: R-praziquantel showed an overall 40% higher area under the plasma drug concentration-time curve (AUC 0→24 ) (R-praziquantel = 3.42; R-polymorph B = 2.05 h*µg/mL) and an overall 30% lower apparent clearance (Cl/F) (R-praziquantel = 70.68 and R-polymorph B = 97.63 (mg)/(µg/mL)/h). Despite the lack of improved activity and PK properties of polymorph B against S. mansoni, here presented; research on pharmaceutical polymorphism remains a valid and cost-effective option for the development of new praziquantel formulations with enhanced properties such as increased solubility and/or dissolution., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Drug discovery for schistosomiasis is still limited to a handful of academic laboratories worldwide, with only a few novel antischistosomal lead compounds being actively researched. Despite recent international mobilization against the disease to stimulate and promote antischistosomal drug discovery, setting up a drug-screening flow with schistosome parasites remains challenging. Whereas numerous different protocols to obtain and cultivate schistosomes have been published, those describing the drug-screening process are scarce, and none gather together parasite cultivation and early drug discovery procedures. To help overcome this hurdle, we provide here a set of integrated methods either adapted from already-published protocols or based on our long-term experience in schistosomiasis research. Specifically, we detail the establishment and maintenance of the complex and several-week-long Schistosoma mansoni life cycle in a laboratory setting, as well as the means of retrieving and culturing the parasites at their relevant life stages. The in vitro and in vivo assays that are performed along the drug-screening cascade are also described. In these assays, which can be performed within 5 d, the effect of a drug is determined by phenotypic assessment of the parasites' viability and morphology, for which stage-specific scoring scales are proposed. Finally, the modalities for testing and evaluating a compound in vivo, constituting a procedure lasting up to 10 weeks, are presented in order to go from in vitro hit identification to the selection of early lead candidates.

Schistosomiasis is a neglected tropical disease, caused by parasitic worms, which affects almost 200 million people worldwide. For over 40 years, chemotherapeutic treatment has relied on the administration of praziquantel, an efficacious drug against schistosomiasis. However, concerns about developing drug resistance require the discovery of novel drug compounds. Currently, the drug-screening process is mostly based on the visual evaluation of drug effects on worm larvae in vitro by a trained operator. This manual process is extremely labor-intensive, has limited throughput, and may be affected by subjectivity of the operator evaluation. In this paper, we introduce a microfluidic platform with integrated electrodes for the automated detection of worm larvae viability using an impedance-based approach. The microfluidic analysis unit consists of two sets of electrodes and a channel of variable geometry to enable counting and size detection of single parasite larvae and the collective evaluation of the motility of the larvae as an unbiased estimator for their viability. The current platform also allows for multiplexing of the analysis units resulting in increased throughput. We used our platform to record size and motility variations of Schistosoma mansoni larvae exposed to different concentrations of mefloquine, a drug with established in vitro antischistosomal properties. The developed platform demonstrates the potential of integrated microfluidic platforms for high-throughput antischistosomal drug screening.

We study the process of decoherence in acoustic black holes. We focus on the ion trap model proposed by Horstmann et al. [Phys. Rev. Lett. 104, 250403 (2010)], but the formalism is general to any experimental implementation. For that particular setup, we compute the decoherence time for the experimental parameters that they proposed. We find that a quantum to classical transition occurs during the measurement, and we propose improved parameters to avoid such a feature. We also study the entanglement between the Hawking-pair phonons for an acoustic black hole while in contact with a reservoir, through the quantum correlations, showing that they remain strongly correlated for small enough times and temperatures.

Geometric phases, arising from cyclic evolutions in a curved parameter space, appear in a wealth of physical settings. Recently, and largely motivated by the need of an experimentally realistic definition for quantum computing applications, the quantum geometric phase was generalized to open systems. The definition takes a kinematical approach, with an initial state that is evolved cyclically but coupled to an environment--leading to a correction of the geometric phase with respect to the uncoupled case. We obtain this correction by measuring the nonunitary evolution of the reduced density matrix of a spin one-half coupled to an environment. In particular we are interested in baths near a quantum phase transition, which are known to induce strong decoherence. The experiments are done with a NMR quantum simulator, where we emulate qualitatively the influence of a critical environment using a simple one-qubit model.

We study the effects of the environment on tunneling in an open system described by a static double-well potential. We describe the evolution of a quantum state localized in one of the minima of the potential at t = 0, in both the limits of high and zero environment temperature. We show that the evolution of the system can be summarized in terms of three main physical phenomena--namely, decoherence, quantum tunneling, and noise-induced activation--and we obtain analytical estimates for the corresponding time scales. These analytical predictions are confirmed by large-scale numerical simulations, providing a detailed picture of the main stages of the evolution and of the relevant dynamical processes.

We study a quantum mechanical toy model that mimics some features of a quenched phase transition. Both by virtue of a time-dependent Hamiltonian or by changing the temperature of the bath we are able to show that even after classicalization has been reached, the system may display quantum behavior again. We explain this behavior in terms of simple nonlinear analysis and estimate relevant time scales that match the results of numerical simulations of the master equation. This opens new possibilities both in the study of quantum effects in nonequilibrium phase transitions and in general time-dependent problems where quantum effects may be relevant even after decoherence has been completed.