MicroRNA-331-3p Suppresses Cervical Cancer Cell Proliferation and E6/E7 Expression by Targeting NRP2

Aberrant expression of microRNAs (miRNAs) is involved in the development and progression of various types of cancers. In this study, we investigated the role of miR-331-3p in cell proliferation and the expression of keratinocyte differentiation markers of uterine cervical cancer cells. Moreover, we evaluated whether neuropilin 2 (NRP2) are putative target molecules that regulate the human papillomavirus (HPV) related oncoproteins E6 and E7. Cell proliferation in the human cervical cancer cell lines SKG-II, HCS-2, and HeLa was assessed using the 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium, inner salt (MTS) assay. Cellular apoptosis was measured using the TdT-mediated dUTP nick end labeling (TUNEL) and Annexin V assays. Quantitative RT-PCR was used to measure the messenger RNA (mRNA) expression of the NRP2, E6, E7, p63, and involucrin (IVL) genes. A functional assay for cell growth was performed using cell cycle analyses. Overexpression of miR-331-3p inhibited cell proliferation, and induced G2/M phase arrest and apoptosis in SKG-II, HCS-2 and HeLa cells. The luciferase reporter assay of the NRP2 3′-untranslated region revealed the direct regulation of NRP2 by miR-331-3p. Gene expression analyses using quantitative RT-PCR in SKG-II, HCS-2, and HeLa cells overexpressing miR-331-3p or suppressing NRP2 revealed down-regulation of E6, E7, and p63 mRNA and up-regulation of IVL mRNA. Moreover, miR-331-3p overexpression was suppressed NRP2 expression in protein level. We showed that miR-331-3p and NRP2 were key effectors of cell proliferation by regulating the cell cycle, apoptosis. NRP-2 also regulates the expression of E6/E7 and keratinocyte differentiation markers. Our findings suggest that miR-331-3p has an important role in regulating cervical cancer cell proliferation, and that miR-331-3p may contribute to keratinocyte differentiation through NRP2 suppression. miR-331-3p and NRP2 may contribute to anti-cancer effects.     

On-off switching properties of one-dimensional photonic crystals consisting of azo-functionalized polymer liquid crystals having different methylene spacers and polyvinyl alcohol

In this paper, photoresponsive behavior of multi-bilayered films having precisely controlled layer thickness prepared by stacking an azo-functionalized polymer liquid crystal, PMAzXAc, and polyvinyl alcohol alternatively, PVA, is described. The multi-bilayered films were found to reflect a light of specific wavelength depending on the layer thickness and refractive index, and showed the reversible change in the reflection intensity by irradiation with visible and UV lights. The change in the reflection intensity was brought about by change in the molecular orientation of PMAzXAc between an out-of-plane orientation and a photo-induced isotropic state, and was strongly dependent on the number of methylene spacer of PMAzXAc linking the azobenzene side group with the acrylate polymer main chain. PMAz6Ac with hexa-methylene spacer showed the largest change in the reflection intensity, while smaller change in the reflection intensity was observed for PMAzXAc having shorter or longer methylene spacer than 6. The effect of the methylene spacers on the photochemical change in the molecular orientation of azobenzene chromophores in the multi-bilayered films will be discussed.     

Burning Rate Augmentation of BAMO Based Propellants

Thermal decomposition and the burning properties of BAMO based propellants with HMX or AN/HMX have been investigated. The heat generated by the azide binder decomposition initiated and accelerated the thermal decomposition of HMX and AN. Ammonium perchlorate (AP) and lead stearate with carbon black significantly altered the mechanisms of the thermal decomposition and the burning properties of the HMX based propellants. AP showed an increase in burning rate with a slight decrease in burning rate pressure exponent. The lead catalyst yielded high value of the burning rate with the lowest pressure exponent. The ammonium dichromate also influenced the mechanisms of the thermal decomposition and the burning properties of the AN/HMX samples. The combination of ammonium dichromate and copper chromite was the most effective on the burning rate augmentation of AN/HMX based propellants. AN sublimed and evaporated from the condensed phase and mainly reacted exothermically in the gas phase HMX and AN/HMX based propellants showed smokeless burning characteristics in the small rocket motor combustion tests.     

Evaluation of drug toxicity with hepatocytes cultured in a micro-space cell culture system

A micro-space cell culture system was recently developed in which cells such as hepatocytes can be cultured and formed into a multicellular three-dimensional (3D) architecture. In this study, we assessed the performance of HepG2 cells cultured in this micro-space cell culture system in a drug toxicity test, and evaluated the effects of micro-space culture on their hepatocyte-specific functions. The micro-space cell culture facilitated the formation of 3D HepG2 cell architecture. HepG2 cells cultured in a micro-space culture plate exhibited increased albumin secretion and enhanced mRNA expression levels of cytochrome P450 (CYP) enzyme compared to those cultured in a monolayer culture. When the cells were exposed to acetaminophen, a hepatotoxic drug, the damage to the HepG2 cells grown in micro-space culture was greater than the damage to the HepG2 cells grown in monolayer culture. In addition, human primary hepatocytes grown in micro-space culture also exhibited increased albumin secretion, enhanced CYP mRNA expression levels and increased sensitivity to acetaminophen compared to those grown in monolayer culture. These results suggest that this micro-space culture method enhances the hepatocyte-specific functions of hepatocytes, including drug-metabolizing enzyme activities, making hepatocytes grown in the micro-space culture system a useful tool for evaluating drug toxicity in vitro.     

Profiling and Imaging of Phospholipids in Brains of Abcd1‐Deficient Mice

ABCD1 is a gene responsible for X‐linked adrenoleukodystrophy (X‐ALD), and is critical for the transport of very long‐chain fatty acids (VLCFA) into peroxisomes and subsequent β‐oxidation. VLCFA‐containing lipids accumulate in X‐ALD patients, although the effect of ABCD1‐deficiency on each lipid species in the central nervous system has not been fully characterized. In this study, each phospholipid and lysophospholipid species in Abcd1‐deficient mice brains were profiled by liquid chromatography‐mass spectrometry. Among the phospholipid and lysophospholipid species that are significantly more enriched in Abcd1‐deficient mice brains, VLCFA were present in 75, 15, 5, 4, and 1 species of phosphatidylcholine, phosphatidylethanolamine, sphingomyelin, lysophosphatidylcholine, and lysophosphatidylethanolamine, respectively. Most VLCFA were incorporated at the sn‐1 position of phosphatidylcholine and phosphatidylethanolamine. Among the phospholipid species that are significantly less enriched in Abcd1‐deficient mice brains, odd‐numbered saturated or mono‐unsaturated fatty acyl moieties are contained in all phosphatidylcholine species. In addition, a number of phosphatidylglycerol, phosphatidylinositol, and phosphatidylserine species contained highly unsaturated fatty acyl moieties. Intriguingly, 44:1 phosphatidylcholine with VLCFA was mainly distributed in the gray matter, such as the cortex, but not in the white matter in the cerebrum and cerebellum. These results show that ABCD1‐deficiency causes metabolic alternation of long‐chain fatty acids and VLCFA. Moreover, our results imply a molecular mechanism for the incorporation of saturated or monounsaturated VLCFA into the sn‐1 position of phospholipids, and also indicate that the distribution of phospholipids with VLCFA may correlate with the development of X‐ALD.     

Guiding and blocking of highly charged ions through a single glass capillary

Highly charged ions produced in an electron beam ion trap, I^q+, q = 10-50, were transmitted through a tapered glass capillary having diameter of at the end. We found that for a particular beam current, there exists an optimum tilting angle of the capillary in which a steady output of ions is observed, while for smaller angles, the ion counts first rise, then gradually decay on a time scale of minutes. In the case of steady transmission, the charge state distribution is found to be slightly towards the lower side.     

Imaging dynamics of charge-auto-organisation in glass capillaries

Multiply charged ion beam transmission through insulating capillaries is today a very active field of research. Thanks to the work of several groups during the last five years, several features of this unexpected process have been evidenced. The open challenge is to understand and control the self-organized charging-up of the capillary walls, which leads finally to the ion transmission. Up to now, the specific charge distribution on the inner surface, as well as the dynamics of the build-up, are still to be understood. While capillaries usually studied are microscopic pore networks etched in different materials, our concern is in macroscopic single capillaries made of glass. With a length of several centimeters and a diameter of a few micrometers at the exit, these capillaries have nevertheless the same aspect ratio as the etched pores (length/diameter ≈ 100). One of the leading goals of this research on single capillaries is to produce multi-charged ion beams with diameters smaller than a micrometer (nano-beams). These glass capillaries offer the opportunity to be used as an ion funnel due to their amazing properties of guiding and focusing highly charged ion beams without altering neither their initial charge state nor the beam emittance (<10⁻³ π mm mrad). However, the understanding of the underlying process is not complete and relies on models assuming charge patches distributed along the capillary and which still need to be tested. We present the first observation imaging the dynamics of the charging-up process in single glass capillaries. During the build-up of the self-organized charge deposition on the capillary walls, the 230 keV Xe²³⁺ transmitted beam is deflected back and forth several times as the outgoing current increases. This is in agreement with the picture of charge patches created sequentially along the capillary and thus deflecting the beam until a stationary state is reached.     

The Association of Cholesterol Uptake and Synthesis with Histology and Genotype in Cortisol-Producing Adenoma (CPA)

Cortisol-producing adenoma (CPA) is composed of clear and compact cells. Clear cells are lipid abundant, and compact ones lipid poor but associated with higher production of steroid hormones. PRKACA mutation (PRKACA mt) in CPA patients was reported to be associated with more pronounced clinical manifestation of Cushing’s syndrome. In this study, we examined the association of histological features and genotypes with cholesterol uptake receptors and synthetic enzymes in 40 CPA cases, and with the quantitative results obtained by gas chromatography-mass spectrometry (GC-MS) analysis in 33 cases to explore their biological and clinical significance. Both cholesterol uptake receptors and synthetic enzymes were more abundant in compact cells. GC-MS analysis demonstrated that the percentage of compact cells was inversely correlated with the concentrations of cholesterol and cholesterol esters, and positively with the activity of cholesterol biosynthesis from cholesterol esters. In addition, hormone-sensitive lipase (HSL), which catalyzes cholesterol biosynthesis from cholesterol esters, tended to be more abundant in compact cells of PRKACA mt CPAs. These results demonstrated that both cholesterol uptake and biosynthesis were more pronounced in compact cells in CPA. In addition, more pronounced HSL expression in compact cells of PRKACA mt CPA could contribute to their more pronounced clinical manifestation.     

Characterization of electrical properties and photosensitivity of SnS thin films prepared by the electrochemical deposition method

Using the electrochemical deposition (ECD) method, we prepared tin sulfide thin films, which are suitable for the absorption layer in solar cells because of its bandgap energy (1 eV). We first optimized pulse-form biasing for ECD by characterizing deposited samples with scanning electron microscope, Auger electron spectroscopy and X-ray diffraction measurements. Then, we investigated the electrical properties of deposited SnS thin films and the properties of contacts with several different metals. Furthermore, we observed the photoconductivity of the films by means of photoelectrochemical measurements. From these results, we confirmed that the SnS thin films show p-type conduction.     

Traversable Region Estimation for Mobile Robots in an Outdoor Image

This paper describes a novel method to estimate appropriate traversable regions from an outdoor scene image. The traversable regions output by the proposed method reflect the common sense of people. For example, a candidate traversable region is “a paved road somewhat distant from the side ditch.” The input to the traversable region estimation is one color image. First, category is assigned to each pixel in the image. The categorization result is then input to the region estimator. Finally, the traversable region are estimated on the input image. An important aspect of this method is the application of two score functions in region estimation process. One score function places high value on categories selected as traversable paths by subjects. The other function places high value on categories that are not selected as traversable regions but are adjacent to categories with traversable paths. A combination of these two functions produces feasible estimation results. The effectiveness of the combined score functions was evaluated by experiments and a questionnaire.