Revolutionising immune research with organoid-based co-culture and chip systems.

The intertwined interactions various immune cells have with epithelial cells in our body require sophisticated experimental approaches to be studied. Due to the limitations of immortalised cell lines and animal models, there is an increasing demand for human in vitro model systems to investigate the microenvironment of immune cells in normal and in pathological conditions. Organoids, which are self-renewing, 3D cellular structures that are derived from stem cells, have started to provide gap-filling tissue modelling solutions. In this review, we first demonstrate with some of the available examples how organoid-based immune cell co-culture experiments can advance disease modelling of cancer, inflammatory bowel disease and tissue regeneration. Then, we argue that to achieve both complexity and scale, organ-on-chip models combined with cutting-edge microfluidics-based technologies can provide more precise manipulation and readouts. Finally, we discuss how genome editing techniques and the use of patient-derived organoids and immune cells can improve disease modelling and facilitate precision medicine. To achieve maximum impact and efficiency, these efforts should be supported by novel infrastructures such as organoid biobanks, organoid facilities, as well as drug screening and host-microbe interaction testing platforms. All these together or in combination can allow researchers to shed more detailed, and often patient-specific, light on the crosstalk between immune cells and epithelial cells in health and disease.

Dynamic shimming in the cervical spinal cord for multi-echo gradient-echo imaging at 3 T.

Obtaining high quality images of the spinal cord with MRI is difficult, partly due to the fact that the spinal cord is surrounded by a number of structures that have differing magnetic susceptibility. This causes inhomogeneities in the magnetic field, which in turn lead to image artifacts. In order to address this issue, linear compensation gradients can be employed. The latter can be generated using an MRI scanner's first order gradient coils and adjusted on a per-slice basis, in order to correct for through-plane ("z") magnetic field gradients. This approach is referred to as z-shimming. The aim of this study is two-fold. The first aim was to replicate aspects of a previous study wherein z-shimming was found to improve image quality in T2*-weighted echo-planar imaging. Our second aim was to improve upon the z-shimming approach by including in-plane compensation gradients and adjusting the compensation gradients during the image acquisition process so that they take into account respiration-induced magnetic field variations. We refer to this novel approach as realtime dynamic shimming. Measurements performed in a group of 12 healthy volunteers at 3 T show improved signal homogeneity along the spinal cord when using z-shimming. Signal homogeneity may be further improved by including realtime compensation for respiration-induced field gradients and by also doing this for gradients along the in-plane axes.

Lung parenchyma and structure visualisation in paediatric chest MRI: a comparison of different short and ultra-short echo time protocols.

AIM: To evaluate image quality acquired at lung imaging using magnetic resonance imaging (MRI) sequences using short and ultra-short (UTE) echo times (TEs) with different acquisition strategies (breath-hold, prospective, and retrospective gating) in paediatric patients and in healthy volunteers. MATERIALS AND METHODS: End-inspiratory and end-expiratory three-dimensional (3D) spoiled gradient (SPGR3D) and 3D zero echo-time (ZTE3D), and 3D UTE free-breathing (UTE3D), prospective projection navigated radial ZTE3D (ZTE3D vnav), and four-dimensional ZTE (ZTE4D) were performed using a 1.5 T MRI system. For quantitative assessment, the contrast-to-noise ratio (CNR) and signal-to-noise ratio (SNR) values were calculated. To evaluate image quality, qualitative scoring was undertaken on all sequences to evaluate depiction of intrapulmonary vessels, fissures, bronchi, imaging noise, artefacts, and overall acceptability. RESULTS: Eight cystic fibrosis (CF) patients (median age 14 years, range 13-17 years), seven children with history of prematurity with or without bronchopulmonary dysplasia (BPD; median 10 years, range 10-11 years), and 10 healthy volunteers (median 32 years, range 20-52 years) were included in the study. ZTE3D vnav provided the most reliable output in terms of image quality, although scan time was highly dependent on navigator triggering efficiency and respiratory pattern. CONCLUSIONS: Best image quality was achieved with prospective ZTE3D and UTE3D readouts both in children and volunteers. The current implementation of retrospective ZTE3D readout (ZTE4D) did not provide diagnostic image quality but rather introduced artefacts over the entire imaging volume mimicking lung pathology.

Deep learning for improving ZTE MRI images in free breathing.

INTRODUCTION: Despite a growing interest in lung MRI, its broader use in a clinical setting remains challenging. Several factors limit the image quality of lung MRI, such as the extremely short T2 and T2* relaxation times of the lung parenchyma and cardiac and breathing motion. Zero Echo Time (ZTE) sequences are sensitive to short T2 and T2* species paving the way to improved "CT-like" MR images. To overcome this limitation, a retrospective respiratory gated version of ZTE (ZTE4D) which can obtain images in 16 different respiratory phases during free breathing was developed. Initial performance of ZTE4D have shown motion artifacts. To improve image quality, deep learning with fully convolutional neural networks (FCNNs) has been proposed. CNNs has been widely used for MR imaging, but it has not been used for improving free-breathing lung imaging yet. Our proposed pipeline facilitates the clinical work with patients showing difficulties/uncapable to perform breath-holding, or when the different gating techniques are not efficient due to the irregular respiratory pace. MATERIALS AND METHODS: After signed informed consent and IRB approval, ZTE4D free breathing and breath-hold ZTE3D images were obtained from 10 healthy volunteers on a 1.5 T MRI scanner (GE Healthcare Signa Artist, Waukesha, WI). ZTE4D acquisition captured all 16 phases of the respiratory cycle. For the ZTE breath-hold, the subjects were instructed to hold their breath in 5 different inflation levels ranging from full expiration to full inspiration. The training dataset consisting of ZTE-BH images of 10 volunteers was split into 8 volunteers for training, 1 for validation and 1 for testing. In total 800 ZTE breath-hold images were constructed by adding Gaussian noise and performing image transformations (translations, rotations) to imitate the effect of motion in the respiratory cycle, and blurring from varying diaphragm positions, as it appears for ZTE4D. These sets were used to train a FCNN model to remove the artificially added noise and transformations from the ZTE breath-hold images and reproduce the original quality of the images. Mean squared error (MSE) was used as loss function. The remaining 2 healthy volunteer's ZTE4D images were used to test the model and qualitatively assess the predicted images. RESULTS: Our model obtained a MSE of 0.09% on the training set and 0.135% on the validation set. When tested on unseen data the predicted images from our model improved the contrast of the pulmonary parenchyma against air filled regions (airways or air trapping). The SNR of the lung parenchyma was quantitatively improved by a factor of 1.98 and the CNR lung- blood, which is indicating the visibility of the intrapulmonary vessels, was improved by 4.2%. Our network was able to reduce ghosting artifacts, such as diaphragm movement and blurring, and enhancing image quality. DISCUSSION: Free-breathing 3D and 4D lung imaging with MRI is feasible, however its quality is not yet acceptable for clinical use. This can be improved with deep learning techniques. Our FCNN improves the visual image quality and reduces artifacts of free-breathing ZTE4D. Our main goal was rather to remove ghosting artifacts from the ZTE4D images, to improve diagnostic quality of the images. As main results of the network, diaphragm contour increased with sharper edges by visual inspection and less blurring of the anatomical structures and lung parenchyma. CONCLUSION: With FCNNs, image quality of free breathing ZTE4D lung MRI can be improved and enable better visualization of the lung parenchyma in different respiratory phases.

Fractional order vs. exponential fitting in UTE MR imaging of the patellar tendon.

PURPOSE: Quantification of the T2∗ relaxation time constant is relevant in various magnetic resonance imaging applications. Mono- or bi-exponential models are typically used to determine these parameters. However, in case of complex, heterogeneous tissues these models could lead to inaccurate results. We compared a model, provided by the fractional-order extension of the Bloch equation with the conventional models. METHODS: Axial 3D ultra-short echo time (UTE) scans were acquired using a 3.0 T MRI and a 16-channel surface coil. After image registration, voxel-wise T2∗ was quantified with mono-exponential, bi-exponential and fractional-order fitting. We evaluated all three models repeatability and the bias of their derived parameters by fitting at various noise levels. To investigate the effect of the SNR for the different models, a Monte-Carlo experiment with 1000 repeats was performed for different noise levels for one subject. For a cross-sectional investigation, we used the mean fitted values of the ROIs in five volunteers. RESULTS: Comparing the mono-exponential and the fractional order T2∗ maps, the fractional order fitting method yielded enhanced contrast and an improved delineation of the different tissues. In the case of the bi-exponential method, the long T2∗ component map demonstrated the anatomy clearly with high contrast. Simulations showed a nonzero bias of the parameters for all three mathematical models. ROI based fitting showed that the T2∗ values were different depending on the applied method, and they differed most for the patellar tendon in all subjects. CONCLUSIONS: In high SNR cases, the fractional order and bi-exponential models are both performing well with low bias. However, in all observed cases, one of the bi-exponential components has high standard deviation in T2∗. The bi-exponential model is suitable for T2∗ mapping, but we recommend using the fractional order model for cases of low SNR.

Liquid-cooled Ti:Sapphire thin disk amplifiers for high average power 100-TW systems.

In this work, numerical heat transfer simulations of direct water-cooled gain modules for thin disk (TD) Ti:Sapphire (Ti:Sa) power amplifiers are presented. By using the TD technique in combination with the extraction during pumping (EDP) method 100-TW class amplifiers operating around 300 W average power could be reached in the future. Single and double-sided cooling arrangements were investigated for several coolant flow velocities. Simulations which upscale the gain module for multiple kilowatts of average power were also performed for large aperture Ti:Sa disks and for multiple disks with several coolant channels.

Autophagy is required for zebrafish caudal fin regeneration.

Regeneration is the ability of multicellular organisms to replace damaged tissues and regrow lost body parts. This process relies on cell fate transformation that involves changes in gene expression as well as in the composition of the cytoplasmic compartment, and exhibits a characteristic age-related decline. Here, we present evidence that genetic and pharmacological inhibition of autophagy - a lysosome-mediated self-degradation process of eukaryotic cells, which has been implicated in extensive cellular remodelling and aging - impairs the regeneration of amputated caudal fins in the zebrafish (Danio rerio). Thus, autophagy is required for injury-induced tissue renewal. We further show that upregulation of autophagy in the regeneration zone occurs downstream of mitogen-activated protein kinase/extracellular signal-regulated kinase signalling to protect cells from undergoing apoptosis and enable cytosolic restructuring underlying terminal cell fate determination. This novel cellular function of the autophagic process in regeneration implies that the role of cellular self-digestion in differentiation and tissue patterning is more fundamental than previously thought.

Current redistribution and generation of kinetic energy in the stagnated Z pinch.

The structure of magnetic fields was investigated in stagnated wire-array Z pinches using a Faraday rotation diagnostic at the wavelength of 266 nm. The distribution of current in the pinch and trailing material was reconstructed. A significant part of current can switch from the main pinch to the trailing plasma preheated by x-ray radiation of the pinch. Secondary implosions of trailing plasma generate kinetic energy and provide enhanced heating and radiation of plasma at stagnation. Hot spots in wire-array Z pinches also provide enhanced radiation of the Z pinch. A collapse of a single hot spot radiates 1%-3% of x-ray energy of the Z pinch with a total contribution of hot spots of 10%-30%.

Investigation of plasma instabilities in the stagnated Z pinch.

High-resolution laser probing diagnostics at a wavelength of 266 nm allow observation of the internal structure and instabilities in dense stagnated Z pinches, typically hidden by trailing material. The internal structure of the 1-MA Z pinch includes strong kink and sausage instabilities, loops, flares, and disruptions. Mid- and small-scale density perturbations develop in the precursor and main pinch. The three-dimensional shape and dynamics of the wire-array Z pinch are predetermined by the initial configuration of the wire array. Cylindrical, linear, and star wire-array Z pinches present different sets of instabilities seeded to the pinch at the implosion stage. Prolonged implosion of trailing mass can enhance x-ray production in wire arrays. Fast plasma motion with a velocity >100 km/s was observed in the Z pinch at stagnation with two-frame shadowgraphy. Development of instabilities in wire arrays is in agreement with three-dimensional magnetohydrodynamic simulations.

Study of the internal structure and small-scale instabilities in the dense Z pinch.

High-resolution laser diagnostics at the wavelength of 266 nm were applied for the investigation of Z pinches at the 1-MA generator. The internal structure of the stagnated Z pinches was observed in unprecedented detail. A dense pinch with strong instabilities was seen inside the column of the trailing plasma. Kink instability, disruptions, and micropinches were seen at the peak of the x-ray pulse and later in time. The three-dimensional structure of the stagnated Z pinch depends on the initial wire-array configuration and implosion scenario. Small-scale density perturbations were found in the precursor plasma and in the stagnated Z pinch. Development of instabilities is in agreement with three-dimensional magnetohydrodynamic simulations.

Measurement of the ionization state and electron temperature of plasma during the ablation stage of a wire-array Z pinch using absorption spectroscopy.

Wire-array plasmas were investigated in the nonradiative ablation stage via x-ray absorption spectroscopy. A laser-produced Sm plasma was used to backlight Al wire arrays. The Sm spectrum was simultaneously observed by two spectrometers: one recorded the unattenuated spectrum and the other the transmission spectrum with 1.45-1.55 keV K-shell absorption lines. Analysis of absorption spectra revealed electron temperature in the range of 10-30 eV and the presence of F-, O-, N- and C-like Al ions in the absorbing plasma. A comparison of this electron temperature with the postprocessed absorption spectra of a 2D MHD simulation yields results in general agreement with the data analysis.

Randomized clinical trial comparing double-lumen tube and EZ-Blocker for single-lung ventilation.

BACKGROUND: In several clinical situations, lung separation and single-lung ventilation (SLV) is essential. In these cases, the double-lumen tube (DLT) is the most widely used device. Bronchial blocker such as Univent or Arndt Blocker serves as an alternative. The EZ-Blocker(®) (EZ; AnaesthetIQ B.V., Rotterdam, The Netherlands) is a new device promising to exceed clinical performance of DLT. The aim of this study was to assess the clinical performance of EZ in comparison with conventional left-sided DLT. METHODS: Forty adult patients undergoing elective thoracic surgery requiring thoracotomy and SLV were included in this study. The patients were randomly assigned to one of two groups: EZ (combined with conventional 7.5 or 8.5 mm single-lumen tube) or DLT (37 or 39 Fr left-sided DLT). Time for intubation procedure and time to verification of the correct position of EZ or DLT using fibreoptic bronchoscopy (FOB) were recorded. After surgery, a thoracic surgeon rated the quality of collapse of the lung (1-3 on a three-level scale). RESULTS: Time for intubation using DLT 85.5 (54.8) s was significantly faster (P<0.001) than using EZ 192 (89.7) s, whereas time for bronchoscopy was not significantly different (P=0.556). Conditions of surgery were rated equally [DLT 1.3 (0.6) vs EZ 1.4 (0.6), P=0.681]. CONCLUSIONS: Although time for intubation was longer with the EZ, the device proved to be an efficient and easy-to-use device. The EZ is a valuable alternative device to conventional DLT. Verification of the correct position of the EZ by FOB seems to be obligatory. This study was registered at http://www.clinicaltrials.gov (identifier: NCT01171560).

Dose response to nitric oxide in adult cardiac surgery patients.

STUDY OBJECTIVE: To determine the dose responsiveness to nitric oxide in adult cardiac surgery patients, especially in those patients with pulmonary hypertension. DESIGN: Prospective, randomized, nonblinded study. SETTING: University teaching hospital. PATIENTS: 62 consecutive cardiac surgery patients demonstrating pulmonary hypertension immediately before induction of anesthesia. INTERVENTIONS: Subjects were assigned by random number allocation to receive one of five doses of inhaled nitric oxide on termination of cardiopulmonary bypass (CBP; i.e., restitution of pulmonary artery flow). Subjects in Group 1 (n = 11) received 10 ppm of inhaled nitric oxide, Group 2 subjects (n = 12) received 20 ppm, Group 3 subjects (n = 12) received 30 ppm, and Group 4 subjects (n = 12) received 40 ppm. The fifth group (n = 15) received no nitric oxide. This fifth group served as a control and was treated with milrinone only. Those patients who were randomized to the milrinone group, had milrinone initiated by bolus administration (50 microg/kg) 15 min before separation from CPB. Milrinone was maintained at 0.5 microg/kg/min in the operating room thereafter. The conduct of anesthesia, surgery, and CBP were controlled. A therapeutic algorithm dictated the use of vasoactive substances for all patients. MEASUREMENTS: Heart rate, mean arterial pressure, pulmonary vascular resistance (PVR), peripheral vascular resistance, cardiac index, and right ventricular ejection fraction were monitored throughout the operative experience. MAIN RESULTS: There were no significant differences found in demographic data, baseline hemodynamic data, surgical treatment, conduct of CBP, or the use of inotropic or vasoactive drugs among the five treatment groups. The percentage decrease in PVR on treatment with nitric oxide as compared to baseline values was not significantly different among the groups (10 ppm = 38%, 20 ppm = 50%, 30 ppm = 44%, 40 ppm = 36%, milrinone = 58%, p = 0.86). CONCLUSIONS: Treatment with nitric oxide was associated with significant reductions in PVR in all groups. Dosages higher than 10 ppm were not associated with greater reductions in pulmonary vascular tone. In view of the fact that nitric oxide-related toxicity is dose-related, doses greater than 10 ppm do not appear to be justified in this patient population.

A prospective comparison of three heat preservation methods for patients undergoing hypothermic cardiopulmonary bypass.

OBJECTIVE: To prospectively compare 3 methods of body heat preservation in patients undergoing surgery requiring the use of hypothermic cardiopulmonary bypass (CPB). DESIGN: Prospective, randomized, and nonblinded. SETTING: University teaching hospital. PARTICIPANTS: Adult cardiac surgery patients (n = 101). INTERVENTIONS: Subjects were randomly assigned to 1 of 3 treatment groups: Group 1 (n = 33) used a fluid-filled warming blanket, group 2 (n = 31) used a heated and humidified breathing circuit, and group 3 (n = 37) used intravenous fluid warmers for the administration of all fluids. Treatments started on separation from CPB and concluded at the end of the intraoperative experience. Anesthetic technique, minute ventilation, conduct of CPB, and room temperature in the operating room were standardized. MEASUREMENTS AND MAIN RESULTS: Blood temperature was measured at its nadir on CPB, on separation from CPB, and just before departure from the operating room. No differences were found among groups for CPB duration, coldest venous temperature on CPB, rewarming time, rate of rewarming, room temperature, or blood temperature on separation from CPB. There were no significant differences found in post-CPB temperature afterdrop among groups. CONCLUSIONS: This study suggests that there is no statistically significant disparity in the effectiveness of these 3 intraoperative heat preservation methods. Ease of use and cost-effectiveness should guide the choice of warming method post-CPB.

A comparison of inhaled nitric oxide and milrinone for the treatment of pulmonary hypertension in adult cardiac surgery patients.

OBJECTIVE: To investigate the relative effects of milrinone and nitric oxide on pulmonary and systemic hemodynamic responses in cardiac surgery patients with a history of pulmonary hypertension. DESIGN: Prospective and randomized. SETTING: University hospital. PARTICIPANTS: Forty-five adult cardiac surgery patients. INTERVENTIONS: Cardiac surgery patients with pulmonary hypertension were randomly assigned to one of three study groups: Group 1 patients (n = 15) were treated with intravenous milrinone on separation from cardiopulmonary bypass, group 2 patients (n = 15) with 20 ppm of inhaled nitric oxide, and group 3 patients (n = 15) with 40 ppm of inhaled nitric oxide. Heart rate, right ventricular ejection fraction, and pulmonary vascular resistance were measured throughout the perioperative period at specific data points. MEASUREMENTS AND MAIN RESULTS: There were no significant differences in demographics, anesthesia, surgery, or baseline hemodynamics among the groups. The group receiving 40 ppm nitric oxide had a significantly higher (p<0.05) right ventricular ejection fraction on arrival in the intensive care unit (40% v. 30% for the milrinone group and 33% for the nitric oxide 20 ppm group). The milrinone group required significantly more phenylephrine in the intensive care unit (p<0.05). CONCLUSIONS: Treatment of pulmonary hypertension in adult cardiac surgery patients with inhaled nitric oxide compared with milrinone is associated with lower heart rates, higher right ventricular ejection fraction, and a lower requirement for treatment with vasopressor agents.

Computational simulations of mitral regurgitation quantification using the flow convergence method: comparison of hemispheric and hemielliptic formulae.

Mitral regurgitation results from the incomplete closure of the mitral valve, and the noninvasive diagnosis of this disease remains an important clinical goal. In this study, steady flow computer simulations were used to evaluate flow convergence method for flow rate estimation. The hemispheric and hemielliptic formulae were compared for accuracy in the presence of complicating factors such as ventricular confinement, orifice shape, and aortic outflow. Results showed that in the absence of aortic outflow and ventricular confinement, there was a plateau zone where the hemispheric formula approximated the true flow rate, independent of orifice shape. However, in the presence of complicating factors such as aortic outflow and ventricular confinement, there was no clear zone where the hemispheric formula could be applied. The hemielliptic formula, however, worked in all cases, regardless of chamber size or magnitude of aortic outflow. Therefore, application of the hemielliptic formula should be considered in future clinical studies.