long term differentiation and functionality of three-dimensional bioprinted primary human hepatocytes: application forengraftment.

In recent decades, 3Dcultures of primary human hepatocytes (PHHs) have been increasingly developed to establish models capable of faithfully mimicking main liver functions. The use of 3D bioprinting, capable of recreating structures composed of cells embedded in matrix with controlled microarchitectures, is an emergent key feature for tissue engineering. In this work, we used an extrusion-based system to print PHH in a methacrylated gelatin (GelMa) matrix.

DMSO-free highly differentiated HepaRG spheroids for chronic toxicity, liver functions and genotoxicity studies.

The liver is essential in the elimination of environmental and food contaminants. Given the interspecies differences between rodents and humans, the development of relevant in vitro human models is crucial to investigate liver functions and toxicity in cells that better reflect pathophysiological processes. Classically, the differentiation of the hepatic HepaRG cell line requires high concentration of dimethyl sulfoxide (DMSO), which restricts its usefulness for drug-metabolism studies.

A New Sleep Staging System for Type III Sleep Studies Equipped With a Tracheal Sound Sensor.

Type III sleep studies record cardio-respiratory channels only. Compared with polysomnography, which also records electrophysiological channels, they present many advantages: they are less expensive, less time-consuming, and more likely to be performed at home. However, their accuracy is limited by missing sleep information.

Generation of proliferating human adult hepatocytes using optimized 3D culture conditions.

Generating the proliferation of differentiated normal adult human hepatocytes is a major challenge and an expected central step in understanding the microenvironmental conditions that regulate the phenotype of human hepatocytes in vitro. In this work, we described optimized 3D culture conditions of primary human hepatocytes (PHH) to trigger two waves of proliferation and we identified matrix stiffness and cell-cell interactions as the main actors necessary for this proliferation. We demonstrated that DNA replication and overexpression of cell cycle markers are modulate by the matrix stiffness while PHH cultured in 3D without prior cellular interactions did not proliferate.

3D culture of HepaRG cells in GelMa and its application to bioprinting of a multicellular hepatic model.

Bioprinting is an emergent technology that has already demonstrated the capacity to create complex and/or vascularized multicellular structures with defined and organized architectures, in a reproducible and high throughput way. Here, we present the implementation of a complex liver model by the development of a three-dimensional extrusion bioprinting process, including parameters for matrix polymerization of methacrylated gelatin, using two hepatic cell lines, Huh7 and HepaRG. The printed structures exhibited long-term viability (28 days), proliferative ability, a relevant hepatocyte phenotype and functions equivalent to or better than those of their 2D counterparts using standard DMSO treatment.

Diagnosis of sleep apnea without sensors on the patient's face.

Study Objectives: Thermistors, nasal cannulas, and respiratory inductance plethysmography (RIP) are the recommended reference sensors of the American Academy of Sleep Medicine (AASM) for the detection and characterization of apneas and hypopneas; however, these sensors are not well tolerated by patients and have poor scorability. We evaluated the performance of an alternative method using a combination of tracheal sounds (TSs) and RIP signals.

Methods: Consecutive recordings of 70 adult patients from the Pays de la Loire Sleep Cohort were manually scored in random order using the AASM standard signals and the combination TS and RIP signals, without respiratory sensors placed on the patient's face.

Tracheal sounds for the scoring of sleep respiratory events in children.

Study Objectives: Oronasal thermistor and nasal cannula are recommended for the scoring of respiratory events (RE) but these sensors are poorly tolerated in children. The aim of the study was to evaluate tracheal sounds (TS) and suprasternal pressure (SSP) for the scoring of RE during sleep in children.

Methods: We compared the detection and characterization of RE by AASM-recommended sensors ("AASM" scoring), with the detection and characterization of RE by the combination of TS and SSP with respiratory inductance plethysmography-sum (TS-RIP scoring), and TS and SSP only (TS scoring).

Automatic identification of sleep and wakefulness using single-channel EEG and respiratory polygraphy signals for the diagnosis of obstructive sleep apnea.

Polysomnography (PSG) is necessary for the accurate estimation of total sleep time (TST) and the calculation of the apnea-hypopnea index (AHI). In type III home sleep apnea testing (HSAT), TST is overestimated because of the lack of electrophysiological sleep recordings. The aim of this study was to evaluate the accuracy and reliability of a novel automated sleep/wake scoring algorithm combining a single electroencephalogram (EEG) channel with actimetry and HSAT signals.

Molecular and tissue alterations of collagens in fibrosis.

The collagen network is altered in fibrotic diseases associated with extracellular matrix (ECM) biosynthesis and remodeling. This mini-review focuses on the quantitative and qualitative modifications of collagens occurring at the molecular and tissue levels in fibrosis. They result from changes in collagen expression, biosynthesis, enzymatic cross-linking and degradation by several protease families.

Determination of extracellular matrix collagen fibril architectures and pathological remodeling by polarization dependent second harmonic microscopy.

Polarization dependence second harmonic generation (P-SHG) microscopy is gaining increase popularity for in situ quantification of fibrillar protein architectures. In this report, we combine P-SHG microscopy, new linear least square (LLS) fitting and modeling to determine and convert the complex second-order non-linear optical anisotropy parameter ρ of several collagen rich tissues into a simple geometric organization of collagen fibrils. Modeling integrates a priori knowledge of polyhelical organization of collagen molecule polymers forming fibrils and bundles of fibrils as well as Poisson photonic shot noise of the detection system.

[1-9-NαC]-crourorb A1 isolated from Croton urucurana latex induces G2/M cell cycle arrest and apoptosis in human hepatocarcinoma cells.

[1-9-NαC]-crourorb A1 is a cyclic peptide isolated from Croton urucurana Baillon latex, found in midwestern Brazil, that has been shown to exert cytotoxic effects against a panel of cancer cell lines. However, the underlying mechanisms responsible for the crourorb A1-induced cytotoxicity in cancer cells remain unknown. In this study, the effects of crourorb A1 on the viability, apoptosis, cell cycle and migration of Huh-7 (human hepatocarcinoma) cells were investigated.

Human T lymphocytes bioactivate heterocyclic aromatic amines by forming DNA adducts.

Heterocyclic aromatic amines (HAA) are formed in cooked meat, poultry and fish but also arise in tobacco smoke and exhaust gases. HAA are potential human carcinogens, which require metabolic activation to exert their genotoxicity. Human tissues can bioactivate HAA to produce reactive intermediates that bind to DNA.

Validation of a Suprasternal Pressure Sensor for Sleep Apnea Classification in Children.

Study Objectives: The recognition and characterization of respiratory events is crucial when interpreting sleep studies. The aim of the study was to validate the sensor, which integrates the recording of respiratory effort by means of suprasternal pressure (SSP), respiratory flow, and snoring for the classification of sleep apneas in children.

Methods: Sleep recordings of 20 children with a median age of 7.

The Disintegrin and Metalloprotease ADAM12 Is Associated with TGF-β-Induced Epithelial to Mesenchymal Transition.

The increased expression of the Disintegrin and Metalloprotease ADAM12 has been associated with human cancers, however its role remain unclear. We have previously reported that ADAM12 expression is induced by the transforming growth factor, TGF-β and promotes TGF-β-dependent signaling through interaction with the type II receptor of TGF-β. Here we explore the implication of ADAM12 in TGF-β-mediated epithelial to mesenchymal transition (EMT), a key process in cancer progression.

Increasing 3D Matrix Rigidity Strengthens Proliferation and Spheroid Development of Human Liver Cells in a Constant Growth Factor Environment.

Mechanical forces influence the growth and shape of virtually all tissues and organs. Recent studies show that increased cell contractibility, growth and differentiation might be normalized by modulating cell tensions. Particularly, the role of these tensions applied by the extracellular matrix during liver fibrosis could influence the hepatocarcinogenesis process.

Linear least square (LLS) method for pixel-resolution analysis of polarization dependent SHG images of collagen fibrils.

A linear least square (LLS) method is proposed to process polarization dependent SHG intensity analysis at pixel-resolution level in order to provide an analytic solution of nonlinear susceptibility χ(2) coefficients and of fibril orientation. This model is applicable to fibrils with identical orientation in the excitation volume. It has been validated on type I collagen fibrils from cell-free gel, tendon and extracellular matrix of F1 biliary epithelial cells.

Integrative analysis of high-throughput RNAi screen data identifies the FER and CRKL tyrosine kinases as new regulators of the mitogenic ERK-dependent pathways in transformed cells.

Background: Cell proliferation is a hallmark of cancer and depends on complex signaling networks that are chiefly supported by protein kinase activities. Therapeutic strategies have been used to target specific kinases but new methods are required to identify combined targets and improve treatment. Here, we propose a small interfering RNA genetic screen and an integrative approach to identify kinase networks involved in the proliferation of cancer cells.

Discoidin domain receptor 1 controls linear invadosome formation via a Cdc42-Tuba pathway.

Accumulation of type I collagen fibrils in tumors is associated with an increased risk of metastasis. Invadosomes are F-actin structures able to degrade the extracellular matrix. We previously found that collagen I fibrils induced the formation of peculiar linear invadosomes in an unexpected integrin-independent manner.

MEK1/2 overactivation can promote growth arrest by mediating ERK1/2-dependent phosphorylation of p70S6K.

The extracellular signal-regulated kinase (ERK)1/2 mitogen-activated protein (MAP) kinase pathway has been involved in the positive and negative regulation of cell proliferation. Upon mitogen stimulation, ERK1/ERK2 activation is necessary for G1- to S-phase progression whereas when hyperactived, this pathway could elicit cell cycle arrest. The mechanisms involved are not fully elucidated but a kinase-independent function of ERK1/2 has been evidenced in the MAPK-induced growth arrest.

The MAPK MEK1/2-ERK1/2 Pathway and Its Implication in Hepatocyte Cell Cycle Control.

Primary cultures of hepatocytes are powerful models in studying the sequence of events that are necessary for cell progression from a G0-like state to S phase. The models mimic the physiological process of hepatic regeneration after liver injury or partial hepatectomy. Many reports suggest that the mitogen-activated protein kinase (MAPK) ERK1/2 can support hepatocyte proliferation in vitro and in vivo and the MEK/ERK cascade acts as an essential element in hepatocyte responses induced by the EGF.

MAPK signaling in cisplatin-induced death: predominant role of ERK1 over ERK2 in human hepatocellular carcinoma cells.

Hepatocellular carcinoma treatment by arterial infusion of cis-diamminedichloroplatinum-II (cisplatin) exhibits certain therapeutic efficacy. However, optimizations are required and the mechanisms underlying cisplatin proapoptotic effect remain unclear. The mitogen-activated protein kinase (MAPK) pathway plays a key role in cell response to cisplatin and the functional specificity of the isoform MAPK/ERK kinase 1 and 2 (MEK1/2) and ERK1/2 could influence this response.

The association of statins and taxanes: an efficient combination trigger of cancer cell apoptosis.

Background: Cancer cell killing might be achieved by the combined use of available drugs. Statins are major anti-hypercholesterolemia drugs, which also trigger apoptosis of many cancer cell types, while docetaxel is a potent microtubule-stabilising agent.

Methods: Here, we looked at the combined effects of lovastatin and docetaxel in cancer cells.

The knock-down of ERCC1 but not of XPF causes multinucleation.

Excision repair cross complementing gene 1 (ERCC1) associated with xeroderma pigmentosum group F (XPF) is a heterodimeric endonuclease historically involved in the excision of bulky helix-distorting DNA lesions during nucleotide excision repair (NER) but also in the repair of DNA interstrand crosslinks. ERCC1 deficient mice show severe growth retardation associated with premature replicative senescence leading to liver failure and death at four weeks of age. In humans, ERCC1 is overexpressed in hepatocellular carcinoma and in the late G1 phase of hepatocyte cell cycle.