Mucosal Immunity Related to CD8+ T Lymphocytes in Children with Helicobacter pylori Gastritis.

Purpose: We investigated the role of CD8+T cells as host immune factors in pediatric patients with Helicobacter pylori gastritis. Methods: Gastric mucosal tissue and blood samples were collected from 39 children, including 11 children with H. pylori infection and 28 children as controls. Anti-CD8 and anti-T-bet antibodies were used for immunohistochemistry of the gastric mucosa. For the cell surface and intracellular staining, peripheral blood mononuclear cells were stained with anti-IL7Rα, anti-CX3CR1, anti-CD8, anti-T-bet, and anti-IFN-γ antibodies. Cytokines of sera such as tumor necrosis factor alpha (TNF-α) and CX3CL1 were analyzed using enzyme- linked immunosorbent assay (ELISA). Results: In the immunohistochemistry of gastric mucosa, the frequency of CD8+ and T-bet+ T cells cells was higher in the H. pylori-positive group than in the control group (26.9± 7.8% vs. 16.9±3.3%, p<0.001; 5.0±2.5% vs. 2.2±0.7%, p=0.001). Between the control and H. pylori-positive groups, the frequency of IL-7RαlowCX3CR1+ CD8+ and T-bet+ INF-γ+ CD8+ T cells were not significantly different between surface and intracellular staining, respectively (40.4±24.0% vs. 38.2±17.8%, p=0.914; 40.4±24.0% vs. 38.2±17.8%, p=0.914). In the ELISA, no significant differences in TNF-α and CX3CL1 concentrations were observed between the control and H. pylori-positive groups (34.3±12.1 pg/mL vs. 47.0±22.6 pg/mL, p=0.114/0.5± 0.1 pg/mL vs. 0.5±0.1 pg/mL, p=0.188). Conclusion: CD8+ T and Th1 cells, which secrete IFN-γ, might play important roles in the mucosal immunity of the stomach in children with H. pylori infection.

Reactive oxygen species-induced changes in glucose and lipid metabolism contribute to the accumulation of cholesterol in the liver during aging.

Aging is a major risk factor for many chronic diseases due to increased vulnerability to external stress and susceptibility to disease. Aging is associated with metabolic liver disease such as nonalcoholic fatty liver. In this study, we investigated changes in lipid metabolism during aging in mice and the mechanisms involved. Lipid accumulation was increased in liver tissues of aged mice, particularly cholesterol. Increased uptake of both cholesterol and glucose was observed in hepatocytes of aged mice as compared with younger mice. The mRNA expression of GLUT2, GK, SREBP2, HMGCR, and HMGCS, genes for cholesterol synthesis, was gradually increased in liver tissues during aging. Reactive oxygen species (ROS) increase with aging and are closely related to various aging-related diseases. When we treated HepG2 cells and primary hepatocytes with the ROS inducer, H2 O2 , lipid accumulation increased significantly compared to the case for untreated HepG2 cells. H2 O2 treatment significantly increased glucose uptake and acetyl-CoA production, which results in glycolysis and lipid synthesis. Treatment with H2 O2 significantly increased the expression of mRNA for genes related to cholesterol synthesis and uptake. These results suggest that ROS play an important role in altering cholesterol metabolism and consequently contribute to the accumulation of cholesterol in the liver during the aging process.

Alkaline Hydrothermal Synthesis, Characterization, and Photocatalytic Activity of TiO2 Nanostructures: The Effect of Initial TiO2 Phase.

One-dimensional (1D) titanate nanostructures were synthesized by hydrothermal route, using commercially available TiO2 (P25) and anatase powders as precursor materials and strong NaOH solution as catalyzer. The prepared titanates were calcined, followed by protonation to produce TiO2 nanostructures having enhanced photocatalytic and photovoltaic properties. The synthesized TiO2 1D nanostructures were characterized using field-emission scanning electron microscope, high-resolution electron microscope, X-ray diffraction analysis, and UV-Vis photospectroscopy to understand the effect of initial TiO2 phase on morphological and crystallographic features, and bandgap. Methylene blue degradation test was applied to evaluate the photoactivity of the products obtained after different stages of the process. The findings indicate that 1D TiO2 nanostructures form by different mechanisms from dissolved aggregates during hydrothermal process, depending on the crystal structure of the initial precursor used. Photocatalytic test results reveal that protonated titanates have considerable adsorption capability, while photocatalytic degradation depends on TiO2 transformation.

Whole-brain serial-section electron microscopy in larval zebrafish.

High-resolution serial-section electron microscopy (ssEM) makes it possible to investigate the dense meshwork of axons, dendrites, and synapses that form neuronal circuits. However, the imaging scale required to comprehensively reconstruct these structures is more than ten orders of magnitude smaller than the spatial extents occupied by networks of interconnected neurons, some of which span nearly the entire brain. Difficulties in generating and handling data for large volumes at nanoscale resolution have thus restricted vertebrate studies to fragments of circuits. These efforts were recently transformed by advances in computing, sample handling, and imaging techniques, but high-resolution examination of entire brains remains a challenge. Here, we present ssEM data for the complete brain of a larval zebrafish (Danio rerio) at 5.5 days post-fertilization. Our approach utilizes multiple rounds of targeted imaging at different scales to reduce acquisition time and data management requirements. The resulting dataset can be analysed to reconstruct neuronal processes, permitting us to survey all myelinated axons (the projectome). These reconstructions enable precise investigations of neuronal morphology, which reveal remarkable bilateral symmetry in myelinated reticulospinal and lateral line afferent axons. We further set the stage for whole-brain structure-function comparisons by co-registering functional reference atlases and in vivo two-photon fluorescence microscopy data from the same specimen. All obtained images and reconstructions are provided as an open-access resource.

Exendin-4 in combination with adipose-derived stem cells promotes angiogenesis and improves diabetic wound healing.

BACKGROUND: Diminished wound healing is a major complication of diabetes mellitus and can lead to foot ulcers. However, there are limited therapeutic methods to treat this condition. Exendin-4 (Ex-4), a glucagon-like peptide-1 receptor agonist, is known to have many beneficial effects on diabetes. In addition, mesenchymal stem cells are known to have wound healing effects. We investigated the effects of Ex-4 in combination with human adipose tissue-derived stem cells (ADSCs) on diabetic wound healing in a diabetic animal model. METHODS: Diabetic db/db (blood glucose levels, >500 mg/dl) or C57BL/6 mice were subjected to wounding on the skin of the back. One day after wounding, each wound received ADSCs (2.5 × 105 cells) injected intradermally around the wound and/or Ex-4 (50 µl of 100 nM Ex-4) topically applied on the wound with a fine brush daily. Wound size was monitored and wound histology was examined. Human endothelial cells and keratinocyte cells were used to assess angiogenesis and vascular endothelial growth factor expression in vitro. RESULTS: Topical administration of Ex-4 or injection of ADSCs resulted in a rapid reduction of wound size in both diabetic and normoglycemic animals compared with vehicle treatment. Histological analysis also showed rapid skin reconstruction in Ex-4-treated or ADSC-injected wounds. A combination of Ex-4 and ADSCs showed a significantly better therapeutic effect over either treatment alone. In vitro angiogenesis assays showed that both Ex-4 and ADSC-conditioned media (CM) treatment improved migration, invasion and proliferation of human endothelial cells. ADSC-CM also increased migration and proliferation of human keratinocytes. In addition, both Ex-4 and ADSC-CM increased the expression of vascular endothelial growth factor. Co-culture with ADSCs increased migration and proliferation of these cells similar to that found after ADSC-CM treatment. CONCLUSIONS: We suggest that Ex-4 itself is effective for the treatment of diabetic skin wounds, and a combination of topical treatment of Ex-4 and injection of ADSCs has a better therapeutic effect. Thus, a combination of Ex-4 and ADSCs might be an effective therapeutic option for the treatment of diabetic wounds, such as foot ulcers.

Diffusion tensor imaging of idiopathic normal-pressure hydrocephalus and the cerebrospinal fluid tap test.

We evaluated relationships between diffusion tensor imaging (DTI) findings and clinical profiles in idiopathic normal-pressure hydrocephalus (INPH) patients, along with differences in DTI parameters between cerebrospinal fluid tap test (CSFTT) responders and non-responders. Fifty-four INPH patients constituted the final group for analysis. Fractional anisotropy (FA), axial diffusivity, radial diffusivity, and mean diffusivity were assessed using atlas-based tract-mapping methods on 20 different fiber tracts. Uncorrected results revealed that CSFTT non-responders, when compared to responders, exhibited lower FA in the left anterior thalamic radiation (ATR), left cingulum-hippocampus (CgH), and left inferior fronto-occipital fasciculus (IFO) and higher axial diffusivity, radial diffusivity, and mean diffusivity in the left CgH and left inferior longitudinal fasciculus (ILF). FA values in the ATR (bilateral), corticospinal tract (right), IFO (bilateral), and ILF (bilateral) were negatively correlated with Unified Parkinson's Disease Rating Scale motor scores. In the right CgH, FA values showed significant positive correlations with Korean-Mini Mental State Examination scores and negative correlations with Clinical Dementia Rating Scale scores. Our findings may suggest a possibility for considering microstructural changes of white matter in patients with ventriculomegaly as potential imaging markers for the prediction of CSFTT responders. Unique patterns of white matter microstructural changes, as measured using DTI, might underlie impairments in distinct symptom domains in patients with INPH.

Abnormal White Matter Integrity in Elderly Patients with Idiopathic Normal-Pressure Hydrocephalus: A Tract-Based Spatial Statistics Study.

We investigated white matter integrity utilizing diffusion tensor imaging in patients with idiopathic normal-pressure hydrocephalus (INPH) who had a positive response to the cerebrospinal fluid tap test and in age- and gender-matched Alzheimer's disease (AD) patients. We enrolled 28 patients with INPH, 28 patients with AD and 20 healthy controls. Tract-based spatial statistics demonstrated that INPH patients had lower fractional anisotropy (FA) in the anterior corona radiate (bilateral), corpus callosum, superior longitudinal fasciculus (bilateral), posterior thalamic radiation (bilateral), external capsule (bilateral) and middle cerebellar peduncle in comparison with the AD and control groups. Volume-of-interest analysis revealed that INPH patients, when compared to the AD and control groups, showed higher mean diffusivity in the anterior corona radiate (bilateral), corpus callosum, superior longitudinal fasciculus (bilateral), posterior thalamic radiation (left), external capsule (bilateral) and middle cerebellar peduncle. And gait dysfunction was significantly correlated with decreased FA in the splenium of the corpus callosum and right external capsule in INPH patients. Our findings may suggest a possibility for considering microstructural changes in white matter integrity in elderly patients as potential imaging markers for differentiation between INPH and AD and may help us understand the potential pathophysiology of gait disturbances associated with INPH.

Unentangled star-shape poly(ε-caprolactone)s as phthalate-free PVC plasticizers designed for non-toxicity and improved migration resistance.

We develop a nontoxic unentangled star-shape poly(ε-caprolactone) (UESPCL) plasticizer with excellent migration resistance for the production of phthalate-free flexible poly(vinyl chloride) (PVC) by means of the ring-opening polymerization of ε-caprolactone, initiated from the multifunctional core, combined with end-capping, and vacuum purification processes. UESPCL is a transparent liquid at room temperature and exhibits unentangled Newtonian behavior because of its extremely short branched segments. UESPCL is biologically safe without producing an acute toxicity response. Torque analysis measurements reveals that UESPCL offers a faster fusion rate and a higher miscibility with PVC compared to a typical plasticizer, di(2-ethylhexyl) phthalate (DEHP). The solid-state (1)H nuclear magnetic resonance (NMR) spectrum reveals that PVC and UESPCL are miscible with an average domain size of less than 8 nm. The flexibility and transparency of the PVC/UESPCL mixture, that is, phthalate-free flexible PVC, are comparable to the corresponding properties of the PVC/DEHP mixture, and the stretchability and fracture toughness of PVC/UESPCL are superior to the corresponding properties of the PVC/DEHP system. Most of all, PVC/UESPCL shows excellent migration resistance with a weight loss of less than 0.6% in a liquid phase, whereas DEHP migrated out of PVC/DEHP into a liquid phase with a weight loss of about 10%.

Use of pupil size to determine the effect of electromagnetic acupuncture on activation level of the autonomic nervous system.

Magnetic fields are widely considered as a method of treatment to increase the therapeutic effect when applied to acupoints. Hence, this study proposes a new method which creates significant stimulation of acupoints by using weak magnetic fields. We conducted this experiment in order to confirm the effect on the activation level of the autonomic nervous system by measuring pupil sizes in cases of stimulation by using manual acupuncture and electromagnetic acupuncture (EMA) at BL15. We selected 30 Hz of biphasic wave form with 570.1 Gauss. To confirm the biopotential by the magnetic flux density occurring in EMA that affected the activation of the autonomic nervous system, we observed the biopotential induced at the upper and the mid left and right trapezius. We observed a significant decrease in pupil size only in the EMA group (p < 0.05), thus confirming that EMA decreased the pupil size through activation of the parasympathetic nerve in the autonomic nervous system. Moreover, we confirmed that the amplitude of the biopotential which was caused by 570.1 Gauss was higher than ±20 µA. Thus, we can conclude that EMA treatment successfully activates the parasympathetic nerve in the autonomic nervous system by inducing a biotransformation by the induced biopotential.

Vivaldi: A Domain-Specific Language for Volume Processing and Visualization on Distributed Heterogeneous Systems.

As the size of image data from microscopes and telescopes increases, the need for high-throughput processing and visualization of large volumetric data has become more pressing. At the same time, many-core processors and GPU accelerators are commonplace, making high-performance distributed heterogeneous computing systems affordable. However, effectively utilizing GPU clusters is difficult for novice programmers, and even experienced programmers often fail to fully leverage the computing power of new parallel architectures due to their steep learning curve and programming complexity. In this paper, we propose Vivaldi, a new domain-specific language for volume processing and visualization on distributed heterogeneous computing systems. Vivaldi's Python-like grammar and parallel processing abstractions provide flexible programming tools for non-experts to easily write high-performance parallel computing code. Vivaldi provides commonly used functions and numerical operators for customized visualization and high-throughput image processing applications. We demonstrate the performance and usability of Vivaldi on several examples ranging from volume rendering to image segmentation.

Electrical impedance characterization of adipose tissue-derived stem cells cultured on indium tin oxide electrodes.

The optical and electrical analysis of the stem cell growth and differentiation using the transparent electrode can be applied to develop new treatment strategies based on stem cell tissue engineering or therapy. In this article, it was investigated whether adipose tissue-derived stem cells (ADSCs) can be cultured on the indium tin oxide electrodes (ITO) and whether the cell growth can be characterized by impedance spectroscopy. From phase contrast microscopy of ADSCs, it was observed that ADSCs adhered and spread on the surface of ITO electrode during cell growth. The measured electrical impedance spectra of the cell-covered electrode were characterized by an equivalent circuit model. By fitting analysis, the resistance and capacitance of ADSCs were extrapolated. The degree of cell coverage on the electrode during cell growth was sensitively detected by monitoring the impedance magnitude.

Glucose-responsive artificial promoter-mediated insulin gene transfer improves glucose control in diabetic mice.

AIM: To investigate the effect of insulin gene therapy using a glucose-responsive synthetic promoter in type 2 diabetic obese mice. METHODS: We employed a recently developed novel insulin gene therapy strategy using a synthetic promoter that regulates insulin gene expression in the liver in response to blood glucose level changes. We intravenously administered a recombinant adenovirus expressing furin-cleavable rat insulin under the control of the synthetic promoter (rAd-SP-rINSfur) into diabetic Lepr(db/db) mice. A recombinant adenovirus expressing ß-galactosidase under the cytomegalovirus promoter was used as a control (rAd-CMV-ßgal). Blood glucose levels and body weights were monitored for 50 d. Glucose and insulin tolerance tests were performed. Immunohistochemical staining was performed to investigate islet morphology and insulin content. RESULTS: Administration of rAd-SP-rINSfur lowered blood glucose levels and normoglycemia was maintained for 50 d, whereas the rAd-CMV-ßgal control virus-injected mice remained hyperglycemic. Glucose tolerance tests showed that rAd-SP-rINSfur-treated mice cleared exogenous glucose from the blood more efficiently than control virus-injected mice at 4 wk [area under the curve (AUC): 21  508.80 ± 2248.18 vs 62  640.00 ± 5014.28, P < 0.01] and at 6 wk (AUC: 29  956.60 ± 1757.33 vs 60  016.60 ± 3794.47, P < 0.01). In addition, insulin sensitivity was also significantly improved in mice treated with rAd-SP-rINSfur compared with rAd-CMV-ßgal-treated mice (AUC: 9150.17 ± 1007.78 vs 11  994.20 ± 474.40, P < 0.05). The islets from rAd-SP-rINSfur-injected mice appeared to be smaller and to contain a higher concentration of insulin than those from rAd-CMV-ßgal-injected mice. CONCLUSION: Based on these results, we suggest that insulin gene therapy might be one therapeutic option for remission of type 2 diabetes.