Silver nanoparticles modified sulfur-containing POSS polymer membrane substrate for adsorption and surface-enhanced Raman scattering analysis of chrysoidine in food samples.

In analysis of complex samples, the stability and sensitivity of surface-enhanced Raman scattering (SERS) substrates may be compromised by matrix interference. To address this issue, a membrane substrate was prepared for fast enrichment, separation, and detection of chrysoidine all-in-one. The silver nanoparticles modified sulfur-containing POSS polymer (AgNPs/POSS-P-S) SERS membrane substrate was fabricated using polyhedral oligomeric silsesquioxane (POSS) as support materials. Through in-situ growth, AgNPs were uniformly modified on POSS-P-S to ensure the stability and SERS activity of the membrane substrate. The enhancement factor of the malachite green was up to 5.3 × 105. By loading the AgNPs/POSS-P-S on membrane, on the other hand, the SERS membrane substrate can also serve as an adsorption medium for separating chrysoidine from sample matrix. Furthermore, the specific sensing mechanism of AgNPs/POSS-P-S for chrysoidine was investigated and a fast, sensitive, and selective method for its quantification was established, with a linear range of 0.010-2.0 mg/L and the limits of detection at 3.7 µg/L. In addition, the SERS method was successfully applied for the analysis of chrysoidine in beverages and chili products with the recoveries in the range of 83.5%-113.4 % and the relative standard deviations in 3.2%-9.0 %. The proposed AgNPs/POSS-P-S membrane based SRES method has great potential for rapid chrysoidine analysis in food samples.

Fgf8 dynamics and critical slowing down may account for the temperature independence of somitogenesis.

Somitogenesis, the segmentation of the antero-posterior axis in vertebrates, is thought to result from the interactions between a genetic oscillator and a posterior-moving determination wavefront. The segment (somite) size is set by the product of the oscillator period and the velocity of the determination wavefront. Surprisingly, while the segmentation period can vary by a factor three between 20 °C and 32 °C, the somite size is constant. How this temperature independence is achieved is a mystery that we address in this study. Using RT-qPCR we show that the endogenous fgf8 mRNA concentration decreases during somitogenesis and correlates with the exponent of the shrinking pre-somitic mesoderm (PSM) size. As the temperature decreases, the dynamics of fgf8 and many other gene transcripts, as well as the segmentation frequency and the PSM shortening and tail growth rates slows down as T-Tc (with Tc = 14.4 °C). This behavior characteristic of a system near a critical point may account for the temperature independence of somitogenesis in zebrafish.

Somite morphogenesis is required for axial blood vessel formation during zebrafish embryogenesis.

Angioblasts that form the major axial blood vessels of the dorsal aorta and cardinal vein migrate toward the embryonic midline from distant lateral positions. Little is known about what controls the precise timing of angioblast migration and their final destination at the midline. Using zebrafish, we found that midline angioblast migration requires neighboring tissue rearrangements generated by somite morphogenesis. The somitic shape changes cause the adjacent notochord to separate from the underlying endoderm, creating a ventral midline cavity that provides a physical space for the angioblasts to migrate into. The anterior to posterior progression of midline angioblast migration is facilitated by retinoic acid-induced anterior to posterior somite maturation and the subsequent progressive opening of the ventral midline cavity. Our work demonstrates a critical role for somite morphogenesis in organizing surrounding tissues to facilitate notochord positioning and angioblast migration, which is ultimately responsible for creating a functional cardiovascular system.

Binding of azobenzene and p-diaminoazobenzene to the human voltage-gated sodium channel Nav1.4.

The activity of voltage-gated ion channels can be controlled by the binding of photoswitches inside their internal cavity and subsequent light irradiation. We investigated the binding of azobenzene and p-diaminoazobenzene to the human Nav1.4 channel in the inactivated state by means of Gaussian accelerated molecular dynamics simulations and free-energy computations. Three stable binding pockets were identified for each of the two photoswitches. In all the cases, the binding is controlled by the balance between the favorable hydrophobic interactions of the ligands with the nonpolar residues of the protein and the unfavorable polar solvation energy. In addition, electrostatic interactions between the ligand and the polar aminoacids are also relevant for p-diaminoazobenzene due to the presence of the amino groups on the benzene moieties. These groups participate in hydrogen bonding in the most favorable binding pocket and in long-range electrostatic interactions in the other pockets. The thermodinamically preferred binding sites found for both photoswitches are close to the selectivity filter of the channel. Therefore, it is very likely that the binding of these ligands will induce alterations in the ion conduction through the channel.

A highly sensitive poly(chrysoidine G)-gold nanoparticle composite based nitrite sensor for food safety applications.

This work demonstrated the development of conducting poly(chrysoidine G) (PCG)-gold nanoparticle (AuNP)-modified fluorine-doped tin oxide (F : SnO2, FTO) film-coated glass electrodes for the sensitive electrochemical detection of nitrite (NO2-). The homogeneously distributed PCG nanoparticle layer was deposited onto the FTO electrode by cyclic voltammetry sweeping. AuNPs were then anchored onto the PCG/FTO electrode by the chemical reduction of pre-adsorbed Au3+ ions. The as-prepared AuNP/PCG/FTO electrode exhibited excellent electrocatalytic activity for the oxidation of NO2- with high sensitivity (approximately 0.63 µA cm-2µM-1) and a low limit of detection (0.095 µM), which is relevant within the normal concentration range of NO2- in human bodily fluids. The AuNP/PCG/FTO sensor showed sufficient reproducibility, repeatability, low interference, and strong recovery for NO2- detection in food samples. These results indicate that the AuNP/PCG nanocomposites have immense potential for the electrochemical detection of other biologically important compounds.

Chitosan/Al2O3-HA nanocomposite beads for efficient removal of estradiol and chrysoidin from aqueous solution.

Alumina, as a support material, was loaded together with chitosan and hydroxyapatite to form chitosan/Al2O3-HA composite beads and was used for estradiol and chrysoidin removal from aqueous solution in the present work. The physicochemical properties of the beads were studied with Scanning Electron Microscopy (SEM), Fourier Transform Infrared Spectrometry (FTIR), thermogravimetric analysis (TGA) and Brunauer-Emmett-Teller (BET) surface area analysis. FTIR spectra confirmed that the chitosan was loaded successfully on Al2O3-HA, and functional groups were immobilized onto the surface of the beads after the synthesis. The adsorption condition including pH, the amount of adsorbent, initial concentration and time were evaluated during the batch experiments. Isotherm data best matched the Langmuir model and the pseudo-second-order model best described the adsorption kinetics. The maximum adsorption capacity was found to be 39.78 mg/g and 23.26 mg/g for estradiol and chrysoidine, respectively. The adsorbed estradiol and chrysoidin were completely eluted from the composite beads with the eluent of 0.1 M H2SO4/MeOH and the regenerated material was used in several cycles without deterioration in its initial performances. This study suggests that the developed composite beads have high potential for the efficient removal estradiol and chrysoidin from aqueous solution.

Standing on the shoulders of giants: two centuries of struggle against meningococcal disease.

Meningococcal disease was first clinically characterised by Gaspard Vieusseux in 1805, and its causative agent was identified by Anton Weichselbaum in 1887, who named it Diplococcus intracellularis menigitidis. From the beginning, the disease was dreaded because of its epidemic nature, predilection for previously healthy children and adolescents, and high mortality. In the last decade of the 19th century, the concept of serum therapy for toxin-related bacterial diseases was identified. This concept was applied to meningococcal disease therapy, in an independent way, by Wilhelm Kolle, August von Wasserman, and Georg Jochmann in Germany, and Simon Flexner in the USA, resulting in the first successful approach for the treatment of meningococcal disease. During the first three decades of the 20th century, serum therapy was the standard treatment for meningococcal disease. With the advent of sulphamides first and then antibiotics, serum therapy was abandoned. The great challenges that infectious diseases medicine is facing and the awaiting menaces in the future in terms of increasing antibiotic resistance, emergence of new pathogens, and re-emergence of old ones without effective therapy, make passive immunotherapy a promising tool. Acknowledging the achievements of our predecessors might teach us some lessons to bring light to our future.

ALDH1A inhibition sensitizes colon cancer cells to chemotherapy.

BACKGROUND: Recent evidence in cancer research, developed the notion that malignant tumors consist of different subpopulations of cells, one of them, known as cancer stem cells, being attributed many important properties such as enhanced tumorigenicity, proliferation potential and profound multidrug resistance to chemotherapy. Several key stem cells markers were identified in colon cancer. In our study we focused on the aldehyde dehydrogenase type 1 (ALDH1) expression in colon cancer-derived cell lines HT-29/eGFP, HCT-116/eGFP and LS-180/eGFP, and its role in the chemoresistance and tumorigenic potential. METHODS: The effect of pharmacological inhibition of ALDH activity by diethylaminobenzaldehyde (DEAB) and also effect of molecular inhibition by specific siRNA was evaluated in vitro in cultures of human colorectal cell lines. The expression level of different isoenzymes of aldehyde dehydrogenase was determined using qPCR. Changes in cell biology were evaluated by expression analysis, western blot and apoptosis assay. The efficiency of cytotoxic treatment in the presence of different chemotherapeutic drugs was analyzed by fluorimetric assay. Tumorigenicity of cells with specific ALDH1A1 siRNA was tested in xenograft model in vivo. RESULTS: Treatment by DEAB partially sensitized the tested cell lines to chemotherapeutics. Subsequently the molecular inhibition of specific isoforms of ALDH by ALDH1A1 or ALDH1A3 siRNA led to sensitizing of cell lines HT-29/eGFP, HCT-116/eGFP to capecitabine and 5-FU. On the model of athymic mice we observed the effect of molecular inhibition of ALDH1A1 in HT-29/eGFP cells by siRNA. We observed inhibition of proliferation of subcutaneous xenografts in comparison to control cells. CONCLUSION: This research, verifies the significance of the ALDH1A isoforms in multidrug resistance of human colorectal cancer cells and its potential as a cancer stem cell marker. This provides the basis for the development of new approaches regarding the treatment of patients with colorectal adenocarcinoma and potentially the treatment of other tumor malignancies.

Interactions of 4,4'-diaminoazobenzene derivatives with telomeric G-quadruplex DNA.

The development of small molecules to stabilize the G-quadruplex structure has garnered significant attention for anticancer drug discovery. Herein, we report the synthesis of several 4,4'-diaminoazobenzene derivatives containing different substituent groups and their ability to bind and stabilize telomeric G-quadruplex DNA. Circular dichroism (CD) spectroscopy was performed to characterize the quadruplex topologies, measure stabilization effects, and evaluate their capabilities for conformational photoregulation. 4,4'-Diaminoazobenzene derivatives were found to moderately stabilize quadruplex structures but not affect conformational photoregulation. This work further develops the design and general understanding of the stabilization effects of small molecules with telomeric G-quadruplex DNA.

Intermittent apnea elicits inactivity-induced phrenic motor facilitation via a retinoic acid- and protein synthesis-dependent pathway.

Respiratory motoneuron pools must provide rhythmic inspiratory drive that is robust and reliable, yet dynamic enough to respond to respiratory challenges. One form of plasticity that is hypothesized to contribute to motor output stability by sensing and responding to inadequate respiratory neural activity is inactivity-induced phrenic motor facilitation (iPMF), an increase in inspiratory output triggered by a reduction in phrenic synaptic inputs. Evidence suggests that mechanisms giving rise to iPMF differ depending on the pattern of reduced respiratory neural activity (i.e., neural apnea). A prolonged neural apnea elicits iPMF via a spinal TNF-α-induced increase in atypical PKC activity, but little is known regarding mechanisms that elicit iPMF following intermittent neural apnea. We tested the hypothesis that iPMF triggered by intermittent neural apnea requires retinoic acid and protein synthesis. Phrenic nerve activity was recorded in urethane-anesthetized and -ventilated rats treated intrathecally with an inhibitor of retinoic acid synthesis (4-diethlyaminobenzaldehyde, DEAB), a protein synthesis inhibitor (emetine), or vehicle (artificial cerebrospinal fluid) before intermittent (5 episodes, ~1.25 min each) or prolonged (30 min) neural apnea. Both DEAB and emetine abolished iPMF elicited by intermittent neural apnea but had no effect on iPMF elicited by a prolonged neural apnea. Thus different patterns of reduced respiratory neural activity elicit phenotypically similar iPMF via distinct spinal mechanisms. Understanding mechanisms that allow respiratory motoneurons to dynamically tune their output may have important implications in the context of respiratory control disorders that involve varied patterns of reduced respiratory neural activity, such as central sleep apnea and spinal cord injury.NEW & NOTEWORTHY We identify spinal retinoic acid and protein synthesis as critical components in the cellular cascade whereby repetitive reductions in respiratory neural activity elicit rebound increases in phrenic inspiratory activity.

Development and optimization of a novel sample preparation method cored on functionalized nanofibers mat-solid-phase extraction for the simultaneous efficient extraction of illegal anionic and cationic dyes in foods.

A simple and efficient three-step sample preparation method was developed and optimized for the simultaneous analysis of illegal anionic and cationic dyes (acid orange 7, metanil yellow, auramine-O, and chrysoidine) in food samples. A novel solid-phase extraction (SPE) procedure based on nanofibers mat (NFsM) was proposed after solvent extraction and freeze-salting out purification. The preferred SPE sorbent was selected from five functionalized NFsMs by orthogonal experimental design, and the optimization of SPE parameters was achieved through response surface methodology (RSM) based on the Box-Behnken design (BBD). Under the optimal conditions, the target analytes could be completely adsorbed by polypyrrole-functionalized polyacrylonitrile NFsM (PPy/PAN NFsM), and the eluent was directly analyzed by high-performance liquid chromatography-diode array detection (HPLC-DAD). The limits of detection (LODs) were between 0.002 and 0.01 mg kg-1, and satisfactory linearity with correlation coefficients (R > 0.99) for each dye in all samples was achieved. Compared with the Chinese standard method and the published methods, the proposed method was simplified greatly with much lower requirement of sorbent (5.0 mg) and organic solvent (2.8 mL) and higher sample preparation speed (10 min/sample), while higher recovery (83.6-116.5%) and precision (RSDs < 7.1%) were obtained. With this developed method, we have successfully detected illegal ionic dyes in three common representative foods: yellow croaker, soybean products, and chili seasonings. Graphical abstract Schematic representation of the process of the three-step sample preparation.

Aldehyde dehydrogenase activity plays no functional role in stem cell-like properties in anaplastic thyroid cancer cell lines.

Recent studies have revealed that aldehyde dehydrogenase (ALDH) is a candidate marker for thyroid cancer stem cells, although its activity is flexible. The goal of this study is to clarify the functional significance of ALDH enzymatic activity on thyroid cancer stem cells properties in anaplastic thyroid cancer cell lines. In vitro sphere formation assay was used to judge the stemness of 4 anaplastic thyroid cancer cell lines (FRO, ACT1, 8505C, and KTC3). Two well-known ALDH inhibitors, N,N-diethylaminobenzaldehyde (DEAB) and disulfiram (DS), were first used. DEAB (50 µM) almost completely suppressed ALDH activity without affecting cell proliferation or spherogenicity. Lack of effect of ALDH suppression on spherogenicity was confirmed using shRNA for ALDH1A3, an ALDH isozyme predominantly expressed in anaplastic thyroid cancer cell lines. In contrast, an ALDH2 inhibitor DS (1 µM) inhibited spherogenicity but did not inhibit ALDH1A3 activity. Based on the recent article from another group reporting the importance of sonic hedgehog (Shh) signaling in ALDH activity and spherogenicity in thyroid cancer, the effects of the Shh inhibitor cyclopamine were also studied. Like DS, cyclopamine (1 µM) decreased spherogenicity but not ALDH activity. Finally, exogenous expression of ALDH1A3 in otherwise ALDH- TPC1 cells (a papillary thyroid cancer cell line) revealed no effect on spherogenicity. In conclusion, we here show no functional role for ALDH activity in thyroid thyroid cancer stem cells properties. That is, ALDH activity and spherogenicity are clearly dissociable. Further understanding of thyroid cancer stem cells biology in thyroid cancers remains necessary for the future development of thyroid thyroid cancer stem cells-targeted therapies.

Transcriptional regulation of genes involved in retinoic acid metabolism in Senegalese sole larvae.

The aim of this study was the characterization of transcriptional regulatory pathways mediated by retinoic acid (RA) in Senegalese sole larvae. For this purpose, pre-metamorphic larvae were treated with a low concentration of DEAB, an inhibitor of RALDH enzyme, until the end of metamorphosis. No differences in growth, eye migration or survival were observed. Nevertheless, gene expression analysis revealed a total of 20 transcripts differentially expressed during larval development and only six related with DEAB treatments directly involved in RA metabolism and actions (rdh10a, aldh1a2, crbp1, igf2r, rarg and cyp26a1) to adapt to a low-RA environment. In a second experiment, post-metamorphic larvae were exposed to the all-trans RA (atRA) observing an opposite regulation for those genes involved in RA synthesis and degradation (rdh10a, aldh1a2, crbp1 and cyp26a1) as well as other related with thyroid- (dio2) and IGF-axes (igfbp1, igf2r and igfbp5) to balance RA levels. In a third experiment, DEAB-pretreated post-metamorphic larvae were exposed to atRA and TTNPB (a specific RAR agonist). Both drugs down-regulated rdh10a and aldh1a2 and up-regulated cyp26a1 expression demonstrating their important role in RA homeostasis. Moreover, five retinoic receptors that mediate RA actions, the thyroid receptor thrb, and five IGF binding proteins changed differentially their expression. Overall, this study demonstrates that exogenous RA modulates the expression of some genes involved in the RA synthesis, degradation and cellular transport through RAR-mediated regulatory pathways establishing a negative feedback regulatory mechanism necessary to balance endogenous RA levels and gradients.

Single and combined genotoxicity effects of six pollutants on THP-1 cells.

The objective of this study was to evaluate the single and combined genotoxic effects of six food pollutants (Chrysoidine G, Sudan I, acid orange II, malachite green, acrylamide, and potassium bromate) on THP-1 cells through comet assay. The results of the single tests indicated that the pollutants increased the percentage of tail DNA (% tail DNA) in a dose-dependent manner. Moreover, the % tail DNA values induced by synthetic colorants (Chrysoidine G, Sudan I, acid orange II, and malachite green) were significantly higher than those by acrylamide or potassium bromate at most concentrations. In the combined tests, Chrysoidine G (422 µmol/L) or acrylamide (400 µmol/L) was mixed with different concentrations of the other five pollutants respectively. In the first combined tests, most mixtures significantly increased the % tail DNA values with the exception of Chrysoidine G and acid orange II. In the second tests, there were no significant differences in the % tail DNA values between the single and combined tests at most cases.

A highly facile and selective Chemo-Paper-Sensor (CPS) for detection of strontium.

Chemosensors have attracted increasing attention for their usefulness on-site detection and monitoring. In this study, we elucidated a novel, facile, and highly selective Chemo-Paper-Sensor (CPS) for detection and monitoring of strontium (Sr(2+)) ions, which means a potent colorimetric sensor based on a Chrysoidine G (CG)-coated paper strip. The CPS for highly selective colorimetric detection of strontium ion was handily analyzed to determine the red-green-blue (RGB) value using portable devices such as desktop digital scanner and mobile phone camera, quantitatively. Interestingly, an orange to dark orange color transition was observed when the aqueous and solid paper colorimetric sensor was introduced to Sr(2+) ion, respectively. It was demonstrated that the value of the signal has a linear relationship with concentrations of the strontium in the 500 ppm to 100 ppb range with a detection limit of 200 ppb. We believe that a newly developed Chemo-Paper-Sensor will be useful in a wide range of sensing applications.

Synergistic action of thermoresponsive and hygroresponsive elements elicits rapid and directional response of a bilayer actuator.

A bilayer actuator composed of thermoresponsive and thermo/hygroresponsive elements is developed, which undergoes fast, directional and autonomous curling with a speed of up to 0.7 m s(-1) and recovers its shape by hydration. In situ tensile testing of the thermal response of individual layers provided insights into the mechanism of actuation of thermo/hygromorphic bilayers.

CdTe quantum dots@luminol as signal amplification system for chrysoidine with chemiluminescence-chitosan/graphene oxide-magnetite-molecularly imprinting sensor.

A sensitive chemiluminescence (CL) sensor based on chemiluminescence resonance energy transfer (CRET) in CdTe quantum dots@luminol (CdTe QDs@luminol) nanomaterials combined with chitosan/graphene oxide-magnetite-molecularly imprinted polymer (Cs/GM-MIP) for sensing chrysoidine was developed. CdTe QDs@luminol was designed to not only amplify the signal of CL but also reduce luminol consumption in the detection of chrysoidine. On the basis of the abundant hydroxy and amino, Cs and graphene oxide were introduced into the GM-MIP to improve the adsorption ability. The adsorption capacities of chrysoidine by both Cs/GM-MIP and non-imprinted polymer (Cs/GM-NIP) were investigated, and the CdTe QDs@luminol and Cs/GM-MIP were characterized by UV-vis, FTIR, SEM and TEM. The proposed sensor can detect chrysoidine within a linear range of 1.0×10(-7) - 1.0×10(-5) mol/L with a detection limit of 3.2×10(-8) mol/L (3δ) due to considerable chemiluminescence signal enhancement of the CdTe quantum dots@luminol detector and the high selectivity of the Cs/GM-MIP system. Under the optimal conditions of CL, the CdTe QDs@luminol-Cs/GM-MIP-CL sensor was used for chrysoidine determination in samples with satisfactory recoveries in the range of 90-107%.

Toxic effects of chrysoidine on human serum albumin: isothermal titration calorimetry and spectroscopic investigations.

Chrysoidine is widely used in industry as a type of azo dye, and is sometimes used illegally as a food additive despite its potential toxicity. Human serum albumin (HSA) is one of the most important proteins in blood plasma and possesses major physiological functions. In the present study, the conformational and functional effects of chrysoidine on HSA were investigated by isothermal titration calorimetry (ITC), multiple spectroscopic methods, a molecular docking study and an esterase activity assay. Based on the ITC results, the binding stoichiometry of chrysoidine to HSA was estimated to be 1.5:1, and was a spontaneous process via a single hydrogen bond. The binding of chrysoidine to HSA induced dynamic quenching in fluorescence, and changes in secondary structure and in the microenvironment of the Trp-214 residue. In addition, the hydrogen bond (1.80 Å) formed between the chrysoidine molecule and the Gln-211 residue. The esterase activity of HSA decreased following the addition chrysoidine due to the change in protein structure. This study details the direct interaction between chrysoidine and HSA at the molecular level and the mechanism for toxicity as a result of the functional changes induced by HSA structural variation upon binding to chrysoidine in vitro. This study provides useful information towards detailing the transportation mechanism and toxicity of chrysoidine in vivo.

[Mutagenic Activity of Four Aminoazo Compounds with Different Carcinogenicity for Rat Liver in the Ames Test].

In this paper in the bacterial Ames test we compared the mutagenicity of four aminoazo compounds, previously studied by other researchers and used for activation of rat liver enzymes, with the carcinogenicity in the rat liver. It was found that in the Ames test they have mutagenic activity, however, this activity does not correlate quantitatively with rat sensitivity to their hepatocarcinogenic action. Thus, the most active carcinogen 3'-methyl-4-dimethylaminoazobenzene causes mutations almost 2.5 times less than weakly carcinogenic ortho-aminoazotoluene, and exactly the same number of mutations as non-carcinogenic N,N-diethyl-4-aminoazobenzene.

[Determination of Microbial Susceptibility to Sulfanilamides by Electrooptic Analysis].

The effect of sulfanilamides (soluble streptocid as an example) on changing of the electrophysical properties (EP) of microbial cells of Escherichia coli XL-1, BL-Ril, Pseudomonasputida C-11 and BA-11 was studied. It was shown that significant changes in the orientation spectra (OS) of the cell suspensions incubated at various concentrations of the sulfanilamide resulted in changing of the electrooptic (EO) signal of the cell suspension at the first five frequencies of the orientation electric field (10-1000 Hz) with the use of soluble streptocid in a concentration of 0.3 mcg/ml. The dynamics of the drug effect on the microbial cells demonstrated a decrease of the EO signal value 5 minutes after the exposure by -59% vs. the control (the cells not exposed to the drug). During the following exposure the EO signal value practically did not change (within 5%). The changes of the OS of the cell suspensions exposed to soluble streptocid significantly differed for the susceptible and resistant strains. Determination of the activity of sulfanilamides by electrooptic analysis of microbial cell suspensions was considered possible. Changing of the microbial suspencion OS under the effect of sulfanilamides can be used as a test on the microbial cell susceptibility to drugs.