Analysis of LINC01314 and miR-96 Expression in Colorectal Cancer Patients via Tissue Microarray-Based Fluorescence In Situ Hybridization.

Methods: A tissue microarray (TMA) containing 76 individual colorectal tumor samples and 28 adjacent normal samples was constructed, and the expression levels of LINC01314 and miR-96 were detected by fluorescence in situ hybridization. Results: The expression levels of both LINC01314 and miR-96 were upregulated in CRC tissues and were associated with vascular metastasis (p < 0.05). A significantly positive correlation was observed between LINC01314 and miR-96 expression in tumor tissues (p < 0.001, r = 0.870). Dominant expression of LINC01314 was a risk factor for both blood vessel invasion (p < 0.05) and poor 5-year survival (p = 0.001, hazard ratio = 4.144). The Kaplan-Meier analysis indicated that patients with LINC01314-dominant expression exhibited worse 5-year survival rates than those with miR-96-dominant expression (p < 0.05). Conclusion: The expression patterns of both LINC01314 and miR-96 may be diagnostic of, and prognostic for, CRC. These findings will facilitate further exploration of the molecular mechanism of lncRNAs in CRC.

Selenomethionine-Dominated Selenium-Enriched Peanut Protein Ameliorates Alcohol-Induced Liver Disease in Mice by Suppressing Oxidative Stress.

Numerous natural compounds are considered as potential therapeutic agents against alcohol-induced liver disease (ALD). Research shows that selenium (Se) has a variety of bioactivities, including liver protecting ability. The present study based on in vitro cell culture models and in vivo mouse models was aimed at examining the contribution of selenomethionine (SeMet)-dominated Se-enriched peanut protein (SePP) to liver protection. SeMet and especially SePP reversed cell viability and cell death, inhibited ethanol induced CYP2E1 activation, decreased reactive oxygen species level, and restored GSH level. Hence, SeMet-dominated SePP alleviates alcohol-induced AML-12 cytotoxicity by suppressing oxidative stress. The p38-dependent mechanism was found to be responsible for SePP-induced Nrf-2 activation. Furthermore, supplementation with SePP and SeMet regulated lipid metabolism and reduced oxidative stress, minimizing liver damage in mice. Selenomethionine-dominated SePP possesses potential therapeutic properties and can be used to treat ALD through the suppression of oxidative stress.

Advantages of Restoring miR-205-3p Expression for Better Prognosis of Gastric Cancer via Prevention of Epithelial-mesenchymal Transition.

PURPOSE: miR-205 is a tumor suppressor and plays an important role in tumor invasiveness. However, the role of miR-205 in human gastric cancer (GC) epithelial-mesenchymal transition (EMT) remains unclear. The aim of this study was to investigate the molecular mechanism of miR-205 in the regulation of EMT in GC invasion. MATERIALS AND METHODS: Quantitative polymerase chain reaction (qPCR) was used to detect the expression of miR-205 in GC. Further, the correlation between the pathological parameters and prognosis of GC was statistically analyzed. A transwell model was used to evaluate the effect of miR-205-3p on the invasion and migration of GC cells. qPCR, western blotting, and luciferase assay were performed to analyze the relationship and target effects between miR-205-3p and the expression of zinc finger electron box binding homologous box 1 (ZEB1) and 2 (ZEB2). RESULTS: We found that the levels of miR-205-3p were significantly lower (P<0.05) in GC tissues than in matched normal tissues. Additionally, the expression of miR-205-3p was related to the tumor invasion depth, lymph node metastasis, lymph node invasion, and tumor, node, metastasis stage. Patients with lower miR-205-3p expression levels in the tumors had a poorer prognosis. The in vitro assays indicated that miR-205-3p could affect the invasion ability and EMT of GC cells by targeting the expression of both ZEB1 and ZEB2. CONCLUSIONS: miR-205-3p promotes GC progression and affects the prognosis of patients by targeting both ZEB1 and ZEB2 to directly influence EMT.

Direct Electrophysiological Mapping of Shape-Induced Affective Perception.

Visual information may convey different affective valences and induce our brain into different affective perceptions. Many studies have found that unpleasant stimuli could produce stronger emotional effects than pleasant stimuli could. Although there has been a notion that triangle is perceived as negative and circle as positive, there has been no systematic study to map the degrees of valence of shapes with different affective perceptions. Here, we employed four shapes (ellipse, triangle, and line-drawn happy and angry faces) to investigate the behavior and electrophysiological responses, in order to systematically study shape-induced affective perception. The reaction time delay and the event-related potential (ERP), particularly the early ERP component, were applied to find the associations with different affective perceptions. Our behavioral results showed that reaction time for angry face was significantly shorter than those for the other three types of stimuli (p < 0.05). In the ERP results, P1, N1, P2, and N2 amplitudes for angry face were significantly larger than those for happy face. Similarly, P1, N1, P2, and N2 amplitudes for triangle were significantly larger than those for ellipse. Particularly, P1 amplitude in the parietal lobe for angry face was the strongest, followed by happy face, triangle, and ellipse. Hence, this work found distinct electrophysiological evidence to map the shape-induced affective perception. It supports the hypothesis that affective strain would induce larger amplitude than affective ease does and strong affective stimuli induce larger amplitude than mild affective stimuli do.

Electrochemical determination of dopamine using a glassy carbon electrode modified with a nanocomposite consisting of nanoporous platinum-yttrium and graphene.

A nanoporous platinum-yttrium alloy (NP-PtY) was fabricated by dealloying ribbons of a PtYAl alloy. Owing to the high porosity and the synergistic effect of Y in the Pt backbone, the NP-PtY exhibits superior structural stability, reproducibility and electrocatalytic activity. An electrochemical sensor was developed for the highly sensitive and selective detection of dopamine (DA) based on the use of a glassy carbon electrode modified with NP-PtY alloy and graphene. The sensor, best operated at 0.16 V vs. SCE, has a linear range covering the 0.9 to 82 µM concentration range, a 0.36 µM detection limit (at S/N = 3), and good selectivity over tyramine, tryptamine, phenethylamine, uric acid, and ascorbic acid. It gave satisfactory results in the determination of DA in spiked samples of urine. Graphical abstract Nanoporous platinum-yttrium alloy (NP-PtY) was fabricated by means of a one-step dealloying process. A glassy carbon electrode modified with the NP-PtY and graphene nanocomposite exhibits a wide linear range and a low detection limit towards dopamine. The sensor has remarkable reproducibility, stability and selectivity.

Facile Synthesis of Nanoporous Pt-Y alloy with Enhanced Electrocatalytic Activity and Durability.

Recently, Pt-Y alloy has displayed an excellent electrocatalytic activity for oxygen reduction reaction (ORR), and is regarded as a promising cathode catalyst for fuel cells. However, the bulk production of nanoscaled Pt-Y alloy with outstanding catalytic performance remains a great challenge. Here, we address the challenge through a simple dealloying method to synthesize nanoporous Pt-Y alloy (NP-PtY) with a typical ligament size of ~5 nm. By combining the intrinsic superior electrocatalytic activity of Pt-Y alloy with the special nanoporous structure, the NP-PtY bimetallic catalyst presents higher activity for ORR and ethanol oxidation reaction, and better electrocatalytic stability than the commercial Pt/C catalyst and nanoporous Pt alloy. The as-made NP-PtY holds great application potential as a promising electrocatalyst in proton exchange membrane fuel cells due to the advantages of facile preparation and excellent catalytic performance.

Phosphorus-doped helical carbon nanofibers as enhanced sensing platform for electrochemical detection of carbendazim.

A combined chemical vapor deposition with high-pressure annealing has been developed for the production of phosphorus-doped helical carbon nanofibers (P-HCNFs). The resulting P-HCNFs have a large specific surface area, well-defined three-dimensional hierarchical helical structure and rapid apparent heterogeneous electron transfer. Based on the high electrocatalytic activity, the P-HCNFs were used to develop an amperometric sensor for carbendazim detection. The experimental results demonstrated that the sensor is promising for the determination of carbendazim in food samples due to the high sensitivity, wide linear range and low detection limit.

Nanocomposite of Au Nanoparticles/Helical Carbon Nanofibers and Application in Hydrogen Peroxide Biosensor.

A combined sol-gel/hydrogen reduction method has been developed for the mass production of helical carbon nanofibers (HCNFs) by the pyrolysis of acetylene at 425 degrees C in the presence of NiO nanoparticles. The synthesized HCNFs were characterized with scanning electron microscopy (SEM), X-ray diffraction (XRD) and high resolution transmission electron microscopy (HRTEM). The helical-structured carbon nanofibers have a large specific surface area and excellent biocompatibility. A novel enzymatic hydrogen peroxide sensor was then successfully fabricated based on the nanocomposites containing HCNFs and gold nanoparticles (AuNPs). The results indicated that the Au/HCNFs nanocomposites exhibited excellent electrocatalytic activity to the reduction of H2O2, offering a wide linear range from 1.0 µM to 3157 µM with a detection limit as low as 0.46 µM. The apparent Michaelis-Menten constant of the biosensor was 0.61 mM. The as-fabricated biosensor showed a rapid and sensitive amperometric response to hydrogen peroxide with acceptable preparation reproducibility and excellent stability. Because of their low cost and high stability, these novel HCNFs represent seem to be a kind of promising biomaterial and may find wide new applications in scopes such as biocatalysis, immunoassay, environmental monitoring and so on.

Visual influence of shapes and semantic familiarity on human sweet sensitivity.

Vision influences taste. It is known that color plays an important role in flavor perception. However, the effect of other features of visual information such as shapes and semantic familiarity of words on the taste perception, particularly on taste sensitivity, is not clear yet. Here we study whether the sweet taste sensitivity of the subjects is affected by such visual inputs. By displaying basic geometric patterns or words with different degrees of semantic familiarity as visual inputs, the subjects rate the hedonic and semantic familiar scores, and taste a series of sucrose solutions, and their sweet sensitivities are accordingly analyzed. Our results show (1) shapes with curvature like circle and ellipse, with higher hedonic scores, increase the sweet sensitivity, whereas angular shapes like square, rectangle, triangle and pentagram do not affect sweet sensitivity; (2) semantic familiar words, with higher hedonic ratings as well, increase sweet sensitivity, whereas unfamiliar words do not affect or even reduce sweet sensitivities.

Heterotrimeric G protein subunit Gγ13 is critical to olfaction.

The activation of G-protein-coupled olfactory receptors on the olfactory sensory neurons (OSNs) triggers a signaling cascade, which is mediated by a heterotrimeric G-protein consisting of α, ß, and γ subunits. Although its α subunit, Gαolf, has been identified and well characterized, the identities of its ß and γ subunits and their function in olfactory signal transduction, however, have not been well established yet. We, and others, have found the expression of Gγ13 in the olfactory epithelium, particularly in the cilia of the OSNs. In this study, we generated a conditional gene knock-out mouse line to specifically nullify Gγ13 expression in the olfactory marker protein-expressing OSNs. Immunohistochemical and Western blot results showed that Gγ13 subunit was indeed eliminated in the mutant mice's olfactory epithelium. Intriguingly, Gαolf, ß1 subunits, Ric-8B and CEP290 proteins, were also absent in the epithelium whereas the presence of the effector enzyme adenylyl cyclase III remained largely unaltered. Electro-olfactogram studies showed that the mutant animals had greatly reduced responses to a battery of odorants including three presumable pheromones. Behavioral tests indicated that the mutant mice had a remarkably reduced ability to perform an odor-guided search task although their motivation and agility seemed normal. Our results indicate that Gαolf exclusively forms a functional heterotrimeric G-protein with Gß1 and Gγ13 in OSNs, mediating olfactory signal transduction. The identification of the olfactory G-protein's ßγ moiety has provided a novel approach to understanding the feedback regulation of olfactory signal transduction pathways as well as the control of subcellular structures of OSNs.

Sweet preference modified by early experience in mice and the related molecular modulations on the peripheral pathway.

The sweet taste is of immense interest to scientists and has been intensively studied during the last two decades. However, the sweet preference modification and the related mechanisms are still unclear. In this study, we try to establish a mice model with manipulated sweet taste preference and explore the involved possible molecular mechanisms. The animals were exposed to acesulfame-K via maternal milk during lactation and the sweet preference tests were carried out when they grew to adulthood. Our results showed that the preference thresholds for sweet taste were increased in adults by early acesulfame-K exposure and the preference ratios for sweet tastants at low or preferred concentrations were decreased. Moreover, by means of qRT-PCR and Western blot, we observed the increased expression of leptin receptor Ob-Rb and downregulation of Gα-gustducin protein in the soft palate. Thereby, the sweet taste sensitivity may be modified by early sweetener experience during lactation. Along the peripheral sweet sensory pathway, the sweet regulator receptors Ob-Rb, CB1 and components of sweet transduction signal Gα-gustducin and T1R2 in both the soft palate and tongue may be cooperatively involved in the plastic development of sweet taste.

Effects of early intraoral acesulfame-K stimulation to mice on the adult's sweet preference and the expression of α-gustducin in fungiform papilla.

Exposure to artificial sweetener acesulfame-K (AK) at early development stages may influence the adult sweet preference and the periphery gustatory system. We observed that the intraoral AK stimulation to mice from postnatal day 4 (P4) to weaning decreased the preference thresholds for AK and sucrose solutions in adulthood, with the preference pattern unchanged. The preference scores were increased in the exposure group significantly when compared with the control group at a range of concentrations for AK or sucrose solution. Meanwhile, more α-Gustducin-labeled fungiform taste buds and cells in a single taste bud were induced from week 7 by the early intraoral AK stimulation. However, the growth in the number of α-Gustducin-positive taste bud or positive cell number per taste bud occurred only in the anterior region, the rostral 1-mm part, but not in the intermediate region, the caudal 4-mm part, of the anterior two-third of the tongue containing fungiform papillae. This work extends our previous observations and provides new information about the developmental and regional expression pattern of α-Gustducin in mouse fungiform taste bud under early AK-stimulated conditions.

Development of an enhanced chemiluminescence ELISA for the rapid detection of acrylamide in food products.

In this work, a polyclonal antibody for acrylamide (AA) was obtained by immunization of rabbits with N-acryloxysuccinimide (NAS) and keyhole limpet hemocyanin (KLH) conjugate. A direct enzyme-linked immunosorbent assay (ELISA) based on this antibody was developed with enhanced chemiluminescent (ECL) detection of AA in food samples. Assay conditions, such as concentrations of antibody and enzyme conjugate and competition time, were optimized. The effects of ionic strength and pH value were investigated. The optimized ECL-ELISA system allowed AA determination in a linear working range of 26.3-221.1 ng mL(-1) with an IC(50) value of 60.6 ng mL(-1) and a limit of detection of 18.6 ng mL(-1). Good recoveries with spiked food samples were obtained with a recovery range from 74.4 to 98.1%, and these results correlated well with those obtained using an HPLC method. This indicates that ECL-ELISA is applicable to the specific detection and routine monitoring of AA in food samples.

Effects of mother's dietary exposure to acesulfame-K in Pregnancy or lactation on the adult offspring's sweet preference.

This study investigates whether mother's exposure to the artificial sweetener acesulfame-K (AK) during pregnancy or lactation affected her adult offspring's sweet preference. It was found that mother's dietary exposure to AK in pregnancy or lactation decreased the preference thresholds for AK and sucrose solutions in the adult offspring, whereas the preference pattern and the most preferred concentration for AK or sucrose solution were unchanged. Furthermore, the preference scores in the exposure groups were increased significantly when compared with the control group at a range of concentrations for AK or sucrose solution. The existence of AK and its dynamic changes within 24 h in amniotic fluid during pregnancy or in mother's milk during lactation after a single oral infusion of AK solution were revealed by the methods of reversed-phase high-performance liquid chromatography and mass spectrometry. Our data suggest that AK can be ingested by the prenatal or postnatal mice through their mother's amniotic fluid or breast milk, producing a long-dated function on the adult's sweet preference.

Facilitation of the development of fungiform taste buds by early intraoral acesulfame-K stimulation to mice.

The gustatory system is susceptible to anatomical modification by postnatal taste stimulations. This study investigated the effects of early intraoral infusion of acesulfame-K solution on the development of fungiform taste buds in mice. It was found that the acesulfame-K infusion increased the number, promoted the maturation, and enlarged the size of taste bud during the postnatal stages, compared with the age-matched controls. This provides fundamental and new information about the development of taste bud under normal and early acesulfame-K-stimulated conditions.

A reliable method to obtain cells of taste buds from fungiform papillae of mice.

Taste buds consist of four kinds of cells which have distinct characteristics and play different roles in recognizing chemical compounds contained in foodstuffs. In this study we describe a procedure for separating viable taste bud cells from the fungiform papillae in mice. After sacrifice with CO(2), the mouse tongue was excised and immediately incubated in collagenase II and dispase II. The epithelium with fungiform papillae was then peeled away from underlying tissue and the anterior one-third region was incubated in a solution of 0.25% trypsin and 0.02M ethylene-diamine-tetraacetic acid (EDTA) for 8-12min. Following incubation, a cell suspension was obtained by mechanical dissociation. Cells in suspension were identified as taste bud cells by their morphology and by immunofluorescence. A 0.25% trypan blue staining demonstrated that nearly 90% of these cells remained viable. Micrographs from scanning electron microscopy illustrated that taste buds were dissociated from the fungiform papillae, while maintaining the integrity of the other part of the dissociated lingual epithelium during incubation. Such a method allows acquisition of viable taste cells and will aid further research in the study of gustatory characteristics.

The relationship between fungiform papillae density and detection threshold for sucrose in the young males.

The aim of this study was to investigate the relationship of fungiform papillae density with taste detection thresholds for sucrose of young male adults. One hundred and eighty two subjects aged 18-23 years (mean age: 21.9 +/- 1.2 years) were included. The densities of fungiform papillae were recorded with the aid of the digital camera, and the taste detection thresholds for sucrose were detected using a modified forced-choice triangle test. The mean density of papillae within all 170 statistic participants was 92.43 +/- 2.64/cm(2), for the 6-mm-diameter stained section of the tongue tip. The average detection threshold was 10.83 +/- 0.24 mmol/l, and the highest and lowest detection thresholds were 19.88 +/- 1.31 and 5.85 +/- 0.43 mmol/l, respectively. Also, an inverse correlation between the fungiform papillae density and the detection threshold was observed.

Quantitative study of taste bud distribution within the oral cavity of the postnatal mouse.

The aim of this study was to investigate the age-related developmental changes of taste bud distribution within the subpopulations at different postnatal ages in the mouse oral cavity. Developmental changes of taste bud distribution on the soft palate, fungiform, foliate and circumvallate papillae in the mouse oral cavity were examined histologically at different postnatal ages. After paraffin embedding, complete serial sections at 10mum thickness were made and stained by routine hematoxylin-eosin staining methods. Digitised images for each section were examined carefully. The existence of a taste pore was used to identify mature taste buds. A two-way analysis of variance (group versus age) was used to analyse differences in taste bud number and characteristics for each of the developmental changes. An independent measures t-test was used to compare two means. No taste buds with pores were observed at birth within circumvallate and foliate papillae. However, 61% of the circumvallate and 58% of the foliate taste buds contained taste pores at 2 weeks after birth. In contrast, at birth, 55% of the taste buds on the soft palate and only 22% of the taste buds within fungiform papillae contained taste pores. Then, the number of mature taste buds (taste buds with pores) increased rapidly 1 week after birth, resulting in 90% of soft palate taste buds and 32% of fungiform taste buds containing taste pores. These results suggests that the earlier maturation of soft palate taste buds compared with the other populations in the oral cavity raises evidence of their significant role in the taste mechanism, especially in the early life of the mouse.

Regional differences in taste bud distribution and alpha-gustducin expression patterns in the mouse fungiform papilla.

The regional differences between distribution patterns and alpha-gustducin expression patterns of the fungiform (FF) taste buds were investigated in the adult mouse, using hematoxylin-eosin staining and immunofluorescence histochemistry on the most anterior region of the tongue (the first millimeter) through the intermediate region of the tongue (the last 1-4 mm). Paraffin sections were prepared from the tip to posterior regions (anterior and intermediate region containing the FF taste buds) of the adult mouse tongue. Results indicate that there were significant regional differences in size and density of taste buds, the cell counts of the single taste bud, and the alpha-gustducin-immunoreactive taste buds between the 2 regions. The taste bud had a characteristic onion-like appearance, and the alpha-gustducin-immunoreactive cell was spindle shaped with elongated processes extending from the base to the pore of the taste buds. These results provide a detailed insight to better understand regional descriptions of mouse taste bud density and size and alpha-gustducin expression with the mouse model.

Differentiation of alpha-gustducin in taste buds of the mouse soft palate and fungiform papillae.

We used alpha-gustducin, a type II taste-cell-specific G protein, to investigate the onset of taste transduction and its relation to the development of the soft palate (SP) and fungiform (FF) papillae taste buds in the mouse. Paraffin wax embedded sections were prepared from the SP and anterior region of the tongue of the mouse from birth until postnatal day (PD) 63. No alpha-gustducin-immunoreactive cells were observed on the day of birth. One day later, alpha-gustducin was immunolocalised in taste buds with pores with a relatively higher frequency recorded in the SP as compared with the FF papillae. The immunoreactive cells were spindle shaped with elongated processes extending from the base to the pore of the taste buds. On PD 7, the number of taste buds containing alpha-gustducin-immunoreactive cells in the SP was three times greater than that of FF papillae. Our results indicate that taste transduction is essentially acquired from the time of birth. Moreover, the onset of taste transduction by the SP taste buds developed earlier than that achieved by taste buds in the FF papillae.