Phase Contrast Research Articles

Random walk and cell morphology dynamics in Naegleria gruberi.

Amoeboid cell movement and migration are wide-spread across various cell types and species. Microscopy-based analysis of the model systems Dictyostelium and neutrophils over the years have uncovered generality in their overall cell movement pattern. Under no directional cues, the centroid movement can be quantitatively characterized by their persistence to move in a straight line and the frequency of re-orientation. Mathematically, the cells essentially behave as a persistent random walker with memory of two characteristic time-scale. Such quantitative characterization is important from a cellular-level ethology point of view as it has direct connotation to their exploratory and foraging strategies. Interestingly, outside the amoebozoa and metazoa, there are largely uncharacterized species in the excavate taxon Heterolobosea including amoeboflagellate Naegleria. While classical works have shown that these cells indeed show typical amoeboid locomotion on an attached surface, their quantitative features are so far unexplored. Here, we analyzed the cell movement of Naegleria gruberi by employing long-time phase contrast imaging that automatically tracks individual cells. We show that the cells move as a persistent random walker with two time-scales that are close to those known in Dictyostelium and neutrophils. Similarities were also found in the shape dynamics which are characterized by the appearance, splitting and annihilation of the curvature waves along the cell edge. Our analysis based on the Fourier descriptor and a neural network classifier point to importance of morphology features unique to Naegleria including complex protrusions and the transient bipolar dumbbell morphologies.

Daily administration of parathyroid hormone slows the progression of basic multicellular units in the cortical bone of the rabbit distal tibia.

Basic Multicellular Units (BMUs) conduct bone remodeling, a critical process of tissue turnover which, if imbalanced, can lead to disease, including osteoporosis. Parathyroid hormone (PTH 1-34; Teriparatide) is an osteoanabolic treatment for osteoporosis; however, it elevates the rate of intra-cortical remodeling (activation frequency) leading, at least transiently, to increased porosity. The purpose of this study was to test the hypothesis that PTH not only increases the rate at which cortical BMUs are initiated but also increases their progression (Longitudinal Erosion Rate; LER). Two groups (n = 7 each) of six-month old female New Zealand white rabbits were both administered 30 μg/kg of PTH once daily for a period of two weeks to induce remodeling. Their distal right tibiae were then imaged in vivo by in-line phase contrast micro-CT at the Canadian Light Source synchrotron. Over the following two weeks the first group (PTH) received continued daily PTH while the second withdrawal group (PTHW) was administrated 0.9 % saline. At four weeks all animals were euthanized, their distal tibiae were imaged by conventional micro-CT ex vivo and histomorphometry was performed. Matching micro-CT datasets (in vivo and ex vivo) were co-registered in 3D and LER was measured from 612 BMUs. Counter to our hypothesis, mean LER was lower (p < 0.001) in the PTH group (30.19 ± 3.01 μm/day) versus the PTHW group (37.20 ± 2.77 μm/day). Despite the difference in LER, osteonal mineral apposition rate (On.MAR) did not differ between groups indicating the anabolic effect of PTH was sustained after withdrawal. The slowing of BMU progression by PTH warrants further investigation; slowed resorption combined with elevated bone formation rate, may play an important role in how PTH enhances coupling between resorption and formation within the BMU. Finally, the prolonged anabolic response following withdrawal may have utility in terms of optimizing clinical dosing regimens.

Absolute value measurement of ion-scale turbulence by two-dimensional phase contrast imaging in Large Helical Device

Absolute value measurements of turbulence amplitude in magnetically confined high-temperature plasmas can effectively explain turbulence-driven transport characteristics and their role in plasma confinements. Two-dimensional phase contrast imaging (2D-PCI) is a technique to evaluate the space-time spectrum of ion-scale electron density fluctuation. However, absolute value measurement of turbulence amplitude has not been conducted owing to the nonlinearity of the detector. In this study, the absolute measurement method proposed in the previous study is applied to turbulence measurement results in the large helical device. As a result, the localized turbulence amplitude at ne = 1.5 × 1019 m-3 is approximately 3.5 × 1015 m-3, which is 0.02% of the electron density. In addition, the evaluated poloidal wavenumber spectrum is almost consistent, within a certain error range, the spectrum being calculated using a nonlinear gyrokinetic simulation. This result is the first to the best of our knowledge to quantitatively evaluate turbulence amplitudes measured by 2D-PCI and compare with simulations.

High energy x-ray Talbot-Lau interferometer employing a microarray anode-structured target source to extend the field of view.

High energy and large field of view (FOV) phase contrast imaging is crucial for biological and even medical applications. Although some works have devoted to achiving a large field of view at high energy through bending gratings and so on, which would be extremely challenging in medical high energy imaging. We analyze the angular shadowing effect of planar gratings in high-energy X-ray Talbot-Lau interferometer (XTLI). Then we design and develop an inverse XTLI coupled with a microarray anode-structured target source, to extend the FOV at high energy. Our experimental results demonstrate the benefit of the source in inverse XTLI and a large FOV of 106.6 mm in the horizontal direction is achieved at 40 keV. Based on this system, experiments of a mouse demonstrate the potential advantage of phase contrast mode in imaging lung tissue. We extend the FOV in a compact XTLI using a microarray anode-structured target source coupled with an inverse geometry, which eliminates grating G0 and relaxes the fabrication difficulty of G2. We believe the established design idea and imaging system would facilitate the wide applications of XTLI in high energy phase contrast imaging.

Open fringes interferograms generation through the phase contrast technique

Phase contrast imaging is a proven technique for sensing optical wavefronts and obtaining qualitative and quantitative insights into phase disturbances. Traditionally, this method involves introducing a phase mask at the Fourier transform plane of the phase object to create a synthetic reference wave that interferes with the analyzed phase disturbance. In this study, we incorporate an intensity-dependent photorefractive material, facilitating the self-induction of a Zernike-type phase filter. We introduce an innovative modification by inducing an angular offset between the synthetic reference wave and the phase distribution of the object. Effective implementation necessitates an illumination area larger than the phase object's support. This method stands out due to its resilience to environmental factors, shared by all common-path interferometers. Furthermore, it leverages open fringe interferograms, consolidating all essential phase information into a single image. This innovation streamlines phase retrieval, eliminating the need for multiple images, and extends applicability beyond small-phase object perturbations. We present results demonstrating the recovered phase distribution using the well-established Takeda technique for open fringe interferograms. This novel approach promises to enhance phase distortion analysis, offering versatility and simplicity in a variety of applications.

Silibinin Prevents TGFβ-Induced EMT of RPE in Proliferative Vitreoretinopathy by Inhibiting Stat3 and Smad3 Phosphorylation.

The purpose of this study was to investigate the effects of silibinin on epithelial-mesenchymal transition (EMT) of retinal pigment epithelial (RPE) and proliferative vitreoretinopathy (PVR) formation, as well as its underlying molecular mechanism. Cellular morphological change and EMT molecular markers were evaluated by using phase contrast imaging, qPCR, and Western blot (WB) to investigate the impact of silibinin on the EMT of ARPE-19 cells. Scratch assay and transwell assay were used to study the effect of silibinin on cell migration. An intravitreally injected RPE-induced rat PVR model was used to assess the effect of silibinin on PVR in vivo. RNA-seq was applied to study the molecular mechanism of silibinin-mediated PVR prevention. Silibinin inhibited TGFβ1-induced EMT and migration of RPE in a dose-dependent manner in vitro. Moreover, silibinin prevented proliferative membrane formation in an intravitreal injected RPE-induced rat PVR model. In line with these findings, RNA-seq revealed a global suppression of TGFβ1-induced EMT and migration-related genes by silibinin in RPEs. Mechanistically, silibinin reduced TGFβ1-induced phosphorylation levels of Smad3 and Stat3, and Smad3 nuclear translocation in RPE. Silibinin inhibits the EMT of RPE cells in vitro and prevents the formation of PVR membranes in vivo. Mechanistically, silibinin inhibits Smad3 phosphorylation and suppresses Smad3 nuclear translocation through the inhibition of Stat3 phosphorylation. These findings suggest that silibinin may serve as a potential treatment for PVR.

Polyploid cell dynamics and death before and after PEG-treatment of a NIH/3T3 derived culture: vinblastine effects on the regulation of cell subpopulations heterogeneity

BackgroundNeoplastic subpopulations can include polyploid cells that can be involved in tumor evolution and recurrence. Their origin can be traced back to the tumor microenvironment or chemotherapeutic treatment, which can alter cell division or favor cell fusion, generating multinucleated cells. Their progeny, frequently genetically unstable, can result in new aggressive and more resistant to chemotherapy subpopulations. In our work, we used NIHs cells, previously derived from the NIH/3T3 line after serum deprivation, that induced a polyploidization increase with the appearance of cells with DNA content ranging from 4 to 24c. This study aimed to analyze the cellular dynamics of NIHs culture subpopulations before and after treatment with the fusogenic agent polyethylene glycol (PEG), which allowed us to obtain new giant polyploid cells. Successively, PEG-untreated and PEG-treated cultures were incubated with the antimicrotubular poison vinblastine. The dynamics of appearance, decrease and loss of cell subpopulations were evaluated by correlating cell DNA content to mono-multinuclearity resulting from cell fusion and division process alteration and to the peculiarities of cell death events.ResultsDNA microfluorimetry and morphological techniques (phase contrast, fluorescence and TEM microscopies) indicated that PEG treatment induced a 4–24c cell increase and the appearance of new giant elements (64–140c DNA content). Ultrastructural analysis and autophagosomal–lysosomal compartment fluorochromization, which allowed us to correlate cytoplasmic changes to death events, indicated that cell depletion occurred through distinct mechanisms: apoptotic death involved 2c, 4c and 8c cells, while autophagic-like death involved intermediate 12–24c cells, showing nuclear (lobulation/micronucleation) and autophagic cytoplasm alterations. Death, spontaneously occurring, especially in intermediate-sized cells, was increased after vinblastine treatment. No evident cell loss by death events was detected in the 64–140c range.ConclusionsPEG-treated NIHs cultures can represent a model of heterogeneous subpopulations originating from cell fusion and division process anomalies. Altogether, our results suggest that the different cell dynamics of NIHs subpopulations can affect the variability of responses to stimuli able to induce cell degeneration and death. Apoptptic, autophagic or hybrid forms of cell death can also depend on the DNA content and ability to progress through the cell cycle, which may influence the persistence and fate of polyploid cell descendants, also concerning chemotherapeutic agent action.

Segmenting Cervical Arteries in Phase Contrast Magnetic Resonance Imaging Using Convolutional Encoder–Decoder Networks

Phase contrast (PC) magnetic resonance imaging (MRI) is a primary method used to quantify blood flow. Cerebral blood flow (CBF) is an important hemodynamic parameter to characterize cerebrovascular and neurological diseases. However, a critical step in CBF quantification using PC MRI is vessel segmentation, which is largely manual, and thus time-consuming and prone to interrater variability. Here, we present encoder–decoder deep learning models to automate segmentation of neck arteries to accurately quantify CBF. The PC-MRI data were collected from 46 Moyamoya (MM) patients and 107 healthy control (HC) participants. Three segmentation U-Net models (Standard, Nested, and Attention) were compared. The PC MRI images were taken before and 15 min after vasodilation. The models were assessed based on their ability to detect the internal carotid arteries (ICAs), external carotid arteries (ECAs), and vertebral arteries (VAs), using the Dice score coefficient (DSC) of overlap between manual and predicted segmentations and receiver operator characteristic (ROC) metric. Analysis of variance, Wilcoxon rank-sum test, and paired t-test were used for comparisons. The Standard U-NET, Attention U-Net, and Nest U-Net models achieved results of mean DSCs of 0.81 ± 0.21, and 0.85 ± 0.14, and 0.85 ± 0.13, respectively. The ROC curves revealed high area under the curve scores for all methods (≥0.95). While the Nested and Attention U-Net architectures accomplished reliable segmentation performance for HC and MM subsets, Standard U-Net did not perform as well in the subset of MM patients. Blood flow velocities calculated by the models were statistically comparable. In conclusion, optimized deep learning architectures can successfully segment neck arteries in PC MRI images and provide precise quantification of their blood flow.

Vacuum ultraviolet-induced surface modification of polyoxymethylene plates for photo-activation bonding

The surface photoactivation technique using vacuum ultra-violet (VUV) light at 172 nm in wavelength has been applied for adhesive-free bonding of polyoxymethylene (POM) plates. The POM surfaces were photochemically modified via VUV light irradiation in dry air conditions, resulting in the formation of oxygen-containing functionalities. The POM plates were successfully bonded with a practical bonding strength below the melting point. The maximum bonding strength was obtained at an irradiation time of 60 min, while a longer irradiation time resulted in lower bonding strength. Surface morphologies of the photoactivated and fractured POM surfaces were observed by a laser scanning microscope. Due to the VUV irradiation, numerous cracks were formed on the POM surfaces. The fractured surface shows that a cohesive failure occurs at the bonding interface. Atomic force microscopy was used to observe the cross-sectional images of the bonding samples. Around 10 μm width, significant phase contrast between the bonding interface and bulk POM indicates the different mechanical properties, due to VUV irradiation. The modified POM surfaces adhered through thermocompression, and the mechanical interlocking and entanglement of POM molecules can result in successful bonding.

Enhancing Contrast of Spatial Details in X-ray Phase-Contrast Imaging through Modified Fourier Filtering

In-line X-ray phase contrast imaging, which is simple to experiment with, provides significantly higher sensitivity, compared to conventional X-ray absorption imaging. The inversion of the relationship between recorded Fresnel diffraction intensity and the phase shift induced by the object is called phase retrieval. The transport of intensity equation (TIE), a simple method of phase retrieval, which is solved by the fast Fourier transform algorithm proposed by Paganin et al., has been widely adopted. However, the existing method suffers from excessive suppression of high-frequency information, resulting in loss of image details after phase retrieval, or insufficient detail contrast, leading to blurry images. Here, we present a straightforward extension of the two-distance FFT-TIE method by modifying the Fourier filter through the use of a five-point approximation to calculate the inverse Laplacian in a discrete manner. Additionally, we utilize a combination of continuous Fourier transform and a four-point approximation to compute the gradient operator. The method is evaluated by simulating samples with a shape similar to the resolution test map and by using a photograph of a dog for further evaluation. The algorithm that incorporates the modified gradient operator and the algorithm that solely utilizes the continuous Fourier transform for gradient computation were compared with the results obtained using the two-distance FFT-TIE method. The comparisons were conducted using the results obtained from two distances from the sample to the detector. The results show that this method improves the contrast of spatial details and reduces the suppression of high spatial frequencies compared to the two-distance FFT-TIE method. Furthermore, in the low-frequency domain, our algorithm does not lose much information compared to the original method, yielding consistent results. Furthermore, we conducted our experiments using carbon rods. The results show that both our method and the FFT-TIE method exhibit low-frequency distortion due to the requirement of close proximity between the absorption maps and the detector. However, upon closer inspection, our proposed method demonstrates superior accuracy in reproducing the finer details of the carbon rod fibers.

Crucial factors for implementing treatment strategies in intractable atonic postpartum haemorrhage: early phase contrast medium extravasation on dynamic CT

To identify the relationship between contrast medium extravasation (CME) on dynamic contrast-enhanced computed tomography (DCT) and clinical information in intractable atonic postpartum haemorrhage (PPH) and its relevance to treatment with uterine artery embolisation (UAE). Of 90 patients who underwent DCT to diagnose PPH, 60 diagnosed with intractable atonic PPH were investigated retrospectively. Maternal background and clinical indicators were analysed to compare the positive and negative factors of early phase CME. Regression analysis was used to investigate the factors associated with CME. The sensitivity, specificity, and positive and negative predictive values of early phase CME for predicting UAE were calculated. Clinical outcomes were compared between the two groups according to the timing of the decision to undergo UAE. Of the 60 patients with intractable atonic PPH, 21 underwent UAE, 20 of whom had early phase CME on DCT. Pre-DCT clinical parameters and clinical indices were not significantly different in presence of early phase CME. Early phase CME was associated with UAE performance, with a sensitivity of 95%, specificity of 87%, positive predictive value of 80%, and negative predictive value of 97%. In cases where UAE was performed after conservative management, there was a significant increase in blood loss and transfusion volume. Early phase CME is not indicated by background factors or clinical findings. UAE is not required when CME cannot be detected in the uterine cavity. If early phase CME is present, UAE should be considered immediately.

Study of Microstructural, Nutritional, and Biochemical Changes in Hulled and Hulless Barley during Storage Using X-ray and Infrared Techniques.

Four varieties of barley (Esma, AC Metacalf, Tradition, and AB Cattlelac), representing four Canadian barley classes, were stored at 17% moisture content (mc) for 8 week. Stored barely was characterized using synchrotron X-ray phase contrast microcomputed tomography, synchrotron X-ray fluorescence imaging, and mid-infrared spectroscopy at the Canadian Light Source, Saskatoon. The deterioration was observed in all the selected varieties of barley at the end of 8 week of storage. Changes due to spoilage over time were observed in the grain microstructure and its nutrient distribution and composition. This study underscores the critical importance of the initial condition of barley grain microstructure in determining its storage life, particularly under unfavorable conditions. The hulled barley varieties showed more deterioration in microstructure than the hulless varieties of barley, where a direct correlation between microstructural changes and alterations in nutritional content was found. All selected barley classes showed changes in the distribution of nutrients (Ca, Fe, K, Mn, Cu, and Zn), but the two-row AC Metcalf variety exhibited more substantial variations in their nutrient distribution (Zn and Mn) than the other three varieties during storage. The two-row class barley varieties showed more changes in biochemical components (protein, lipids, and carbohydrates) than the six-row class varieties.

Effect of nozzle geometry features on the nozzle internal flow and cavitation characteristics based on X-ray dynamic imaging

The nozzle geometry has a significant impact on the nozzle internal flow, which affects the fuel spray atomization and combustion of internal combustion engine. In this study, the internal geometry and flow characteristics of the nozzle was studied visually by using X-ray phase contrast imaging technique. The results indicate that the angle between the orifice wall and sac significantly influences the formation and development of cavitation in the orifice. A numerical model including the nozzle internal flow and the spray near-field characteristics was established to provide a more detailed description of how the inlet included angle affects the orifice's internal flow. It has been found that the hydraulic flip width increases with the decrease of the inlet included angle. The symmetry of cavitation distribution is greater when the inlet included angles on the left and right sides of the orifice are equal. When the inlet included angles on the left and right sides of the orifice are different, as the right inlet included angle increases, the cavitation strength near the orifice right-side wall decreases, and the hydraulic flip width to decrease, which causes leading to an increase in the effective flow area of the orifice.

Fabrication and In Vitro Biological Assay of Thermo-Mechanically Tuned Chitosan Reinforced Polyurethane Composites.

The progressive trend of utilizing bioactive materials constitutes diverse materials exhibiting biocompatibility. The innovative aspect of this research is the tuning of the thermo-mechanical behavior of polyurethane (PU) composites with improved biocompatibility for vibrant applications. Polycaprolactone (CAPA) Mn = 2000 g-mol-1 was used as a macrodiol, along with toluene diisocyanate (TDI) and hexamethylene diisocyanate (HMDI), to develop prepolymer chains, which were terminated with 1,4 butane diol (BD). The matrix was reinforced with various concentrations of chitosan (1-5 wt %). Two series of PU composites (PUT/PUH) based on aromatic and aliphatic diisocyanate were prepared by varying the hard segment (HS) ratio from 5 to 30 (wt %). The Fourier-transformed infrared (FTIR) spectroscopy showed the absence of an NCO peak at 1730 cm-1 in order to confirm polymer chain termination. Thermal gravimetric analysis (TGA) showed optimum weight loss up to 500 °C. Dynamic mechanical analysis (DMA) showed the complex modulus (E*) ≥ 200 MPa. The scanning electron microscope (SEM) proved the ordered structure and uniform distribution of chain extender in PU. The hemolytic activities were recorded up to 15.8 ± 1.5% for the PUH series. The optimum values for the inhibition of biofilm formation were recorded as 46.3 ± 1.8% against E. coli and S. aureus (%), which was supported by phase contrast microscopy.

Range and variability of CBF in young adults: PC‐MRI and ASL studies

AbstractSeveral studies have compared phase contrast magnetic resonance imaging (PC‐MRI) with arterial spin labeling (ASL) in the same cohort of subjects. However, these earlier comparisons included subjects across a wide age range, which can contribute to increased intersubject variability. We compared the range and variability of CBF based on PC‐MRI and ASL in a group of young subjects. We also explored whether the comparison between PC‐MRI and ASL was dependent on sex. Thirty young subjects (17 females) were recruited in this study. The CBF from PC‐MRI (CBFPC) was calculated as the total blood flow normalized to brain parenchyma mass. The CBF from ASL (CBFASL) was quantified based on the perfusion kinetic model. The ratio between CBFASL and CBFPC was calculated. Females exhibited significantly higher CBF using both PC‐MRI (p &lt; 0.05) and ASL (p &lt; 0.005) compared with males. The disparity in CBF between sexes was more prominent in ASL measurements. A significant correlation was observed between CBFPC and CBFASL values (r = 0.47, p &lt; 0.01), and CBFASL tended to be lower than CBFPC values (p &lt; 0.001). This difference between CBFASL and CBFPC was more pronounced in males, as evidenced by the smaller CBFASL/CBFPC ratio in males (p &lt; 0.05). The measurement of global CBF with PC‐MRI and ASL are correlated, but CBFASL exhibits lower values than CBFPC. As the underestimation of CBFASL is prominent in males, ASL amplifies the sexual dimorphism in CBF compared with PC‐MRI.

Screening of postoperative cerebral hyperperfusion syndrome in moyamoya disease: a three-dimensional pulsed arterial-spin labeling magnetic resonance imaging approach

IntroductionMoyamoya disease (MMD) is associated with a risk of postoperative cerebral hyperperfusion syndrome (CHS) after revascularization surgery. This study aimed to explore the feasibility of using three-dimensional pulsed arterial spin labeling (3D PASL) and phase contrast (PC) magnetic resonance imaging (MRI) for predicting CHS occurrence in patients with MMD before revascularization surgery.MethodsOverall, 191 adult patients (207 hemispheres) with MMD who underwent combined revascularization surgery were included in this study. Preoperative 3D PASL-MRI and PC-MRI were performed before surgery. The PASL-MRI data were analyzed using SPM12. Patient clinical information, average flow, and preoperative cerebral blood flow (CBF) were compared between the non-CHS and CHS groups.ResultsAmong the patients, 45 (21.74%) developed CHS after revascularization surgery. No significant differences were noted in age, sex, clinical symptoms, hypertension, diabetes, surgical side, or history of revascularization surgery between the non-CHS and CHS groups. However, the average flow in the superficial temporal artery was significantly lower in the CHS group than in the non-CHS group (p &amp;lt; 0.05). Furthermore, 11 clusters of preoperative CBF values were significantly greater in the CHS group than in the non-CHS group [p &amp;lt; 0.05, false discovery rate (FDR) corrected]. A significant correlation was also observed between the preoperative time-to-flight MR angiography (MRA) scores and CBF values in patients with MMD (p &amp;lt; 0.05).ConclusionCompare patients with lower preoperative CBF and higher preoperative average flow in the STA, patients with higher preoperative CBF and lower preoperative average flow in the STA are more likely to develop postoperative CHS Preoperative PASL-MRI and PC-MRI examinations may help to screen patients at high risk of developing CHS after revascularization surgery.

Miniaturized microscope for non-invasive imaging of leukocyte-endothelial interaction in human microcirculation

We present a miniature oblique back-illumination microscope (mOBM) for imaging the microcirculation of human oral mucosa, enabling real-time, label-free phase contrast imaging of individual leukocytes circulating in the bloodstream, as well as their rolling and adhesion on vascular walls—the initial steps in leukocyte recruitment that is a hallmark of inflammation. Using the mOBM system, we studied the leukocyte-endothelial interactions in healthy and locally inflamed tissue and observed drastic changes in leukocyte movement (velocity and displacement profile). Our findings suggest that real-time imaging of leukocyte dynamics can provide new diagnostic insights (assessment of inflammation, temporal progression of disease, evaluation of therapeutic response, etc.) that are not available using conventional static parameters such as cell number and morphology.

Point excess solute: A new metric for quantifying solute segregation in atom probe tomography datasets including application to naturally aged solute clusters in Al-Mg-Si-(Cu) alloys

Accurate, repeatable and quantitative analysis of nanoscale solute clustering in atom probe tomography (APT) datasets is a complex challenge which is made more difficult by the positional uncertainty and lack of absolute resolution inherent to the technique. In this work a new method, the point excess solute, is introduced for quantifying solute segregation in datasets with limited spatial resolution. This new method is based on measuring the matrix concentration using a dataset sampling method. We show the new method can accurately reproduce the values expected from synthetic datasets a priori and when the dataset spatial resolution and or phase contrast is too low for accurate quantification this is observable. The method is then applied to naturally aged solute clusters in the Al-Mg-Si-Cu system. Datasets were collected with a range of natural ageing times from 8 min to 76 weeks. The formation of the solute clusters is shown to be unaffected by the Cu content of the alloy.

Effect of Wnt/β-catenin signaling pathway in neural differentiation of human bone marrow mesenchymal stem cells

To explore the effect of basic fibroblast growth factor (bFGF), epidermal growth factor (EGF), and the combination of bFGF and EGF in the neural differentiation of human bone marrow mesenchymal stem cells (hBMSCs), and the role of Wnt/β-catenin signaling pathway in this process. The identified 4th-generation hBMSCs were divided into five groups according to different induction conditions, namely control group (group A), EGF induction group (group B), bFGF induction group (group C), EGF and bFGF combined induction group (group D), and EGF, bFGF, and Dickkopf-related protein 1 (DKK-1) combined induction group (group E). After 7 days of continuous induction, the cell morphology was observed by inverted fluorescence phase contrast microscopy, levels of genes that were related to neural cells [Nestin, neuron-specific enolase (NSE), microtubule-associated protein 2 (MAP-2), and glial fibrillary acidic protein (GFAP)] and key components of the Wnt/β-catenin signaling pathway (β-catenin and Cyclin D1) were detected by RT-PCR, and the levels of proteins that were related to neural cells (Nestin and GFAP) as well as genes that were involved in Wnt/β-catenin signaling pathway [β-catenin, phosphorylation β-catenin (P-β-catenin), Cytoplasmic β-catenin, and Nuclear β-catenin] were explored by cellular immunofluorescence staining and Western blot. When compared to groups A and B, the typical neuro-like cell changes were observed in groups C-E, and most obviously in group D. RT-PCR showed that the relative expressions of Nestin, NSE, and MAP-2 genes in groups C-E, the relative expressions of GFAP gene in groups D and E, the relative expression of NSE gene in group B, the relative expressions of β-catenin gene in groups C and D, and the relative expressions of Cyclin D1 gene in groups B-D significantly increased when compared with group A ( P<0.05). Compared with group E, the relative expressions of Nestin, NSE, MAP-2, GFAP, β-catenin, and CyclinD1 genes significantly increased in group D ( P<0.05); compared with group C, the relative expression of Nestin gene in group D significantly decreased ( P<0.05), while NSE, MAP-2, and GFAP genes significantly increased ( P<0.05). The cellular immunofluorescence staining showed that the ratio of NSE- and GFAP-positive cells significantly increased in groups C-E than in group A, in group D than in groups C and E ( P<0.05). Western blot assay showed that the relative expression of NSE protein was significantly higher in groups C and D than in group A and in group D than in groups C and E ( P<0.05). In addition, the relative expression of GFAP protein was significantly higher in groups C-E than in group A and in group D than in group E ( P<0.05). Besides, the relative expressions of β-catenin, Cytoplasmic β-catenin, Nuclear β-catenin, and the ratio of Nuclear β-catenin to Cytoplasmic β-catenin were significantly higher in groups C and D than in group A and in group D than in group E ( P<0.05), whereas the relative expression of P-β-catenin protein was significantly lower in groups C and D than in group A and in group D than in group E ( P<0.05). Different from EGF, bFGF can induce neural differentiation of hBMSCs. In addition, EGF can enhance the hBMSCs neural differentiation of bFGF, while the Wnt/β-catenin signaling pathway may play a positive regulatory role in these processes.

Anti-inflammatory and cytotoxicity assay of Cardamom-based nutraceuticals in comparison with 1,8 -Cineol

Aim: To investigate anti-inflammatory, cytotoxic effects and viability of Recovereez- 51% 1,8-Cineol and 1,8-Cineol (natural), and to determine safety for both of the compounds and its use in new product range of nutraceuticals.&#x0D; Methodology: Cell line was divided into two groups; one was treated with Recovereez(R) and another one with 1,8-Cineol(C). The ELISA assay method was used to measure the COX, LOX, MPO, NO, and nitrate levels. Cellular viability was performed by observing cells under the inverted phase contrast microscope, followed by MTT assay.&#x0D; Results: R&amp;C showed a reduction in levels of inflammatory markers such as IL-6, IL-1β, TNF-α, LOX, MPO, COX, iNOS, and nitrate levels. The inhibition as observed in the levels of COX and LOX enzymes were 65%±0.1 and 59%±0.3µg/ml, and 72%±0.3 and 67%±0.4µg/ml for R&amp;C, respectively. iNOS production showed inhibition by 70%±0.3 and 65%±0.8µg/ml and NO production were reduced by 193 and 209.5µg/ml by R&amp;C, respectively. Again, there was a decrease in the activity of MPO by 0.35 and 0.39U/ml for R&amp;C, respectively. The results were significant with p&lt;0.001 compared to the Diclofenac sodium. Cell viability of 60% was observed upon treating L929 cells with R&amp;C.&#x0D; Conclusion: R&amp;C exerted an anti-inflammatory effect on RAW 264.7 cells without exerting cytotoxicity in a dose-dependent manner. Therefore, Recovereez could be used as an alternative treatment or to prevent inflammatory diseases. Moreover, R&amp;C are expected to treat different types of cancer. The findings could potentially lead to the discovery of safe and effective bioactive compounds that can prevent or cure the occurrence of cancerous cells.&#x0D; Keywords: dietary supplements, nutraceutical, COX, anti-inflammation, cancer