Development of a visible-light-active-NiTiO3 coating for the efficient removal of the persistent herbicide 2,6-dichlorobenzamide (BAM) from drinking water.

In the present research work, the photocatalytic evaluation of NiTiO3 nanoparticles immobilized on glass plates by the spin-coating procedure was carried out in the degradation of the recalcitrant herbicide 2,6-dichlorobenzamide (BAM). The concentrations of Ni employed to synthesize NiTiO3 nanoparticles were 1 wt% (1TESNi) and 2 wt% (2TESNi). The stability of coatings was evaluated by several washings and thermal treatments, which were verified by UV-vis analyses. The morphology of the coatings was studied by scanning electron microscopy (SEM-EDS). The coatings displayed thickness values of 1.35 and 2.56 µm for TiO2 and 1TESNi, respectively. The crystalline phases of the coatings were analyzed by X-ray diffraction (XRD), confirming the presence of NiTiO3 and other phases related to TiO2. The bandgap of 1TESNi, compared with the bare TiO2, was reduced from 2.96 to 2.40 eV as a consequence of Ni addition. The TiO2, 1TESNi and 2TESNi coatings were evaluated in the photodegradation of BAM using visible-light for 240 min. The highest effectiveness was displayed by the 1TESNi coating, obtaining degradation of 92.56% after 240 min. Also, the photocatalytic efficiency of the 1TESNi coating was only reduced 1.99% after 3 reuse cycles in the BAM degradation. The scavenger tests revealed that the main oxidizing species involved in the reaction were the •OH- and •O2- radicals. The 1TESNi coating showed the highest photocatalytic efficiency because of its absorption in the visible-light region, valuable surface area and electronic charge separation. Thus, these advantageous features guarantee that NiTiO3 coatings are an efficient method for degrading recalcitrant herbicides from drinking water using a practical way to recover and reuse photocatalysts.

Novel polypropylene-TiO2:Bi spherical floater for the efficient photocatalytic degradation of the recalcitrant 2,4,6-TCP herbicide.

In this work, spherical photocatalytic floaters were fabricated by depositing TiO2:Bi (TBi) particles on polypropylene (PP) spheres (recycled from beer cans). These particles were deposited on the sphere (TBi-sphere) by the spray coating technique and evaluated their performance for the photocatalytic degradation of 2,4,6-trichlorophenol (2,4,6-TCP) herbicide. SEM images demonstrated that the BTi powders consisted in conglomerated grains with sizes of 20-80 nm and the analysis by X-ray diffraction confirmed the presence of rutile and anatase phases in the BTi. The photocatalytic experiments showed that the TBi and TBi-sphere produced maximum degradation of 90 and 97% for 2,4,6-TCP, respectively, after 4 h under UV-Vis light. The photocatalytic powders/composites were reused 3 times and the loss of degradation efficiency was 3 and 16% for the TBi powder and TBi-sphere, respectively. This means that the TBi-sphere is more stable for the continuous degradation of the 2,4,6-TCP contaminant. The TiO2:Bi powder was compared with the commercial TiO2 (P25) and found that the TiO2:Bi powder had higher light absorption (≈42%) and higher surface area (≈105%) than the P25. Therefore, the degradation percentage for the 2,4,6-TCP was 52% higher in the sample doped with Bi. Also, scavenger experiments were carried out and found that the main oxidizing agents produced for the degradation of 2,4,6-TCP were •OH- radicals and •O2- anions. Other species such as h+ were also produced at lower amount. Hence, our results demonstrated that spherical/floatable photocatalytic composites are a viable option to remove herbicide residuals from the water, which is of interest in water-treatment-plants.

An eco-friendly and sustainable support of agave-fibers functionalized with graphene/TiO2:SnO2 for the photocatalytic degradation of the 2,4-D herbicide from the drinking water.

In this research, we evaluated the photocatalytic performance of biodegradable composites for the removal of the 2,4-Dichlorophenoxyacetic acid (2,4-D) herbicide. The composite was composed by agave fibers (AgF), graphene-microplates (GM) and titanium dioxide TiO2/SnO2 (TSn) nanoparticles (NPs) and was named TSn + AgF/GM. Both, the TSn NPs and the GM were deposited on the AgF using the Dip-coating method. According to the analysis by X-Ray Diffraction (XRD), the crystalline phase for the TiO2 and SnO2 was anatase and tetragonal-rutile, respectively. The Scanning Electron Microscopy (SEM) images demonstrated that the AgF were completely saturated by the GM (which had average dimensions of 15 µm × 22 µm) and by conglomerations of TSn NPs with average size of 642 nm. The TSn NPs and the TSn + AgF/GM composite were evaluated for the photocatalytic degradation of the 2,4-D herbicide under ultraviolet-visible (UV-Vis) light and found a maximum degradation of 98.4 and 93.7% (after 4 h) for the TSn NPs and the TSn + AgF/GM composite, respectively. Reuse cycles were also performed and the degradation percentage decreased by 13.1% and by 7.8% (after 3 cycles of reuse) when the TSn NPs and the TSn + AgF/GM composite are employed, respectively. Scavenger experiments were also carried out and found that the oxidizing agents are mainly produced in the order of: •OH>•O2- > h+; then, the main oxidizing agents generated during the photocatalytic reaction were the hydroxyl radicals. Thus, the photocatalytic system studied in this work for the degradation of 2,4-D could pave the way for the development of new eco-friendly/floatable photocatalysts, which can be applied in wastewater-treatment plants.

Hydrogen embrittlement through the formation of low-energy dislocation nanostructures in nanoprecipitation-strengthened steels.

Hydrogen embrittlement is shown to proceed through a previously unidentified mechanism. Upon ingress to the microstructure, hydrogen promotes the formation of low-energy dislocation nanostructures. These are characterized by cell patterns whose misorientation increases with strain, which concomitantly attracts further hydrogen up to a critical amount inducing failure. The appearance of the failure zone resembles the "fish eye" associated to inclusions as stress concentrators, a commonly accepted cause for failure. It is shown that the actual crack initiation is the dislocation nanostructure and its associated strain partitioning.

Replacing xylene with n-heptane for paraffin embedding.

In standard histological technique, aromatic solvents such as xylene and toluene are used as clearing agents between ethanol dehydration and paraffin embedding. In addition, these solvents are used for de-waxing paraffin sections. Unfortunately, these solvents are harmful and therefore adequate substitutes would be useful. We suggest the use of n-heptane as a convenient substitute for xylene. Paraffin sections of rat tissues processed with n-heptane and stained with hematoxylin-eosin or Masson's trichrome showed proper embedment, well preserved morphology and excellent staining.

Binding of cationic dyes to DNA: distinguishing intercalation and groove binding mechanisms using simple experimental and numerical models.

Simple methods for predicting intercalation or groove binding of dyes and analogous compounds with double stranded DNA are described. The methods are based on a quantitative assessment of the aspect (width to length) ratio of the dyes. The procedures were validated using a set of 38 cationic dyes of varied chemical structures binding to well oriented DNA fibers and assessing binding orientation by linear dichroism and polarized fluorescence. We demonstrated that low aspect ratio dyes bound by intercalation, whereas more rod-like dyes were groove binders. Some problems that result and possible applications are discussed briefly.

Cytological detection of Wolbachia in squashed and paraffin embedded insect tissues.

Wolbachia localization in situ is essential for accurate analysis of the infection and its consequences. Whole cell hybridization is proposed as an easy and rapid method for detecting Wolbachia cells in paraffin embedded tissues or testis squashes of Chorthippus parallelus (Orthoptera). Wolbachia is found in whole gonads and other adjacent tissues. A higher bacterial density, however, is observed in ovarioles and testis. Small independent bacteria with strictly cytoplasmic distribution are displayed. Bacterial density differences among individuals are also revealed.

Apoptosis and proliferation of fibroblasts during postnatal skin development and scleroderma in the tight-skin mouse.

Tight-skin (Tsk) is a dominant gene mutation that causes a fibrotic skin disease in mice, similar to human scleroderma. Both conditions are characterized by increased numbers of dermal fibroblasts containing high levels of procollagen mRNA. Whether this fibroblast population arises from fibroblast growth or fibroblast transcriptional activation is debated. Proliferation and apoptosis of fibroblasts of normal and Tsk mice were studied in skin sections before, at onset, and in established fibrosis. Tissues sections were immunostained with proliferating cell nuclear antigen (PCNA) as proliferation marker. Apoptosis was investigated by in situ end-labeling of fragmented DNA and nuclear staining with propidium iodide. The expression of the apoptosis inhibitor Bcl-2 was investigated by immunohistochemistry. We demonstrate differences in fibroblast proliferation and apoptosis related to postnatal skin growth and development. Neonatal skin exhibits the highest levels of proliferation and apoptosis in fibroblasts. In contrast, low proliferation and absence of apoptosis characterizes adult fibroblasts. Skin fibroblasts express Bcl-2 only in newborns, and at other ages Bcl-2 was restricted to epithelial cells. Our results also suggest that neither increased fibroblast proliferation nor defective apoptosis accounts for the fibrotic phenotype of Tsk. Therefore, transcriptional activation of extracellular matrix genes appears more relevant in the pathogenesis of Tsk fibrosis.

Influence of inorganic salts on the staining reaction of eosinophil leukocyte granules by anionic dyes.

The use of salts as competing agents in staining solutions allows to evaluate to what extent ionic interactions take place between microscopical substrates and cationic or anionic dyes. We have employed this method to study the staining reaction of horse eosinophil leucocyte granules by the anionic dyes nuclear fast red, naphthol yellow S, eosin Y, indigocarmin, acid fuchsin, alizarin red S, orange G, and Evans blue in the presence of NaCl (from 0.015 to 2 mol/l). Different values of "minimal electrolyte concentration" (the least amount of salt which reduces the staining intensity) were found for these dyes. Comparative observations using ammonium sulphate as competing salt showed that it is less effective than NaCl. Staining of eosinophil granules by anionic dyes could be not suppressed at 2 mol/l NaCl, which indicates that in addition to electrostatic forces, other non-ionic interactions are also responsible for some staining reaction of these acidophilic structures.

DNA fluorescence induced by polymethine cation pyrvinium binding.

Pyrvinium is a polymethine cation which shows interesting fluorescence emission and DNA binding properties. In diluted aqueous solution, pyrvinium pamoate induced a bright yellow fluorescence in kinetoplast DNA from Trypanosoma cruzi epimastigotes as well as in chicken erythrocyte nuclei under a wide range of excitations. No fading was observed after mounting in suitable media. Spectroscopic studies on pyrvinium solutions revealed bathochromic and hypochromic shifts in the absorption spectrum of its complex with DNA. A striking enhancement of pyrvinium fluorescence was found in solvents of high viscosity or after binding to DNA. Experimental results and the chemical structure of pyrvinium allow us to suggest that the minor groove of adenine-thymine DNA regions could be the specific binding site for this new DNA fluorochrome.

Fluram induces species-dependent C and G bands in mammalian chromosomes, revealing heterogeneous distribution of chromosomal proteins.

Fluram (Fluorescamine; 4-phenylspiro(furan-2(3H),1'-phthalan)-3,3'-dione) is a fluorogenic reagent, which permits the detection of primary amines by forming highly fluorescent pyrrolinone derivatives. This reagent has been used on methanol-acetic acid fixed metaphase chromosomes of mouse and man and proved to be very effective in differentiating chromosome regions in both genomes. Mouse centromeric heterochromatin is highly reactive, showing intense fluorescence in all centromeric regions, whereas human chromosomes show no fluorescence in such regions. In addition, a G-like banding pattern is also obtained in both types of chromosomes. The differential reactivity of each chromosome region showed by this method demonstrates a heterogeneous distribution of chromosome proteins, resulting in a chromosome banding pattern, which is in this case species dependent.

Alkaline hydrolysis/methylation-acetylation: a new technique for ultrastructural DNA cytochemistry.

A new technique for the visualization of DNA-containing structures in electron microscopy is described. Samples of glutaraldehyde-fixed bone marrow from rats were subjected to alkaline hydrolysis to remove RNA and the phosphate of phosphoproteins, followed by a combined blockage of protein carboxyl and amino groups through methylation-acetylation. After uranyl acetate staining of epoxy-embedded ultrathin sections, chromatin from all cell types showed a highly selective and intense electron opacity. Staining methods for DNA were also positive in semithin sections. This simple procedure could be very useful in ultrastructural cytochemistry of DNA and chromatin.

Indium (III)-hematoxylin as a staining and contrasting agent for light and electron microscopy.

A deeply violet indium (III)-hematoxylin complex is formed when indium trichloride is added to an aqueous solution of oxidized hematoxylin. Treatment of glutaraldehyde fixed and Araldite embedded sections of rat seminiferous tubules with indium-hematoxylin revealed a definite staining and contrasting pattern. Semithin sections showed chromatin and nucleoli in violet-blue. Under the electron microscope, chromatin, nucleoli, ribosomes, synaptonemal complexes, chromatoid bodies, membranous components, and microtubules from sperm tails presented high electron opacity, while the acrosome and basement membrane appeared with a lower contrast. This performed indium-hematoxylin complex, which shows an absorption peak at lambda = 560 nm with shoulders at about lambda = 440 and 400 nm, could be valuable as a new staining and electron contrasting agent.

X-ray microanalysis of toluidine blue stained chromosomes: a quantitative study of the metachromatic reaction of chromatin.

The stoichiometry of metachromatic staining of chromatin by toluidine blue was investigated in isolated metaphase chromosomes from L929 cells using X-ray microanalysis. Microspectrophotometric measurements revealed that a hypsochromic shift (from 595 to 570 nm) occurs in toluidine blue stained chromosomes in relation to the staining solution. Under the electron microscope, stained chromosomes. After toluidine blue staining, X-ray microanalysis of chromosomes revealed a large increase for sulphur counts and a considerable increase for Fe and Cu counts, while the signal of Mg, Ca, Cl, K and Zn was reduced. After subtraction of the intrinsic sulphur signal, S/P ratios of 0.82--for euchromatic arms--and 0.85--for centromeric heterochromatin--were obtained. They are considered representative of dye/DNA phosphate ratios. These results indicate the occurrence of a nearly stoichiometric binding of toluidine blue to chromatin DNA and suggest that an external dye stacking is responsible for the metachromatic staining of metaphase chromosomes.

Selective digestion of mouse chromosomes with restriction endonucleases. II. X-ray microanalysis of HaeIII-treated chromosomes.

We used X-ray microanalysis to study the changes induced in mouse metaphase chromosomes as a result of digestion with the restriction endonuclease HaeIII. The phosphorus X-ray signal was used as a marker for DNA and the sulfur signal for protein. Calcium, iron, copper, and zinc were also detected. HaeIII induced a loss of phosphorus from both the centromeres and chromosome arms, but the losses in the arms were much greater. These changes were accompanied by an increase in the electron density of the centromeres and a reduction in that of the arms. No reduction in the sulfur signal in either arms or centromeres occurred as a result of HaeIII digestion. Except for calcium, which showed only a moderate reduction, the inorganic ions exhibited very large losses as a result of HaeIII digestion. The differentiation of chromosome arms and centromeres as a result of HaeIII digestion is therefore not simply due to differential loss of DNA but also involves structural reorganization of the chromatin, as shown by electron microscopy. This reorganization does not involve loss of proteins but may be correlated with changes in the amounts of inorganic ions known to be involved in chromatin condensation.

New fluorescence reactions in DNA cytochemistry. 1. Microscopic and spectroscopic studies on nonrigid fluorochromes.

Some nonrigid DNA-binding antibiotics and fluorochromes that recognize adenine-thymine (AT) sequences are widely applied in biomedical research, but the microscopic use, spectral characteristics and DNA binding modes of other similar compounds have been overlooked or scarcely explored. After treatment with thioflavine T, auramine O and G, curcumin, bis-aminophenyl-oxadiazol, berenil and distamycin A, a bright DNA-dependent fluorescence reaction was found in the chromatin of interphase nuclei, meiotic and polytene chromosomes, spermatozoa heads and kinetoplasts of Trypanosoma cruzi epimastigotes. Nucleoli and basophilic cytoplasm showed low or no fluorescence; the highest emission occurred in the AT-rich kinetoplast DNA. When bound to DNA or in the presence of alpha-cyclodextrin and viscous solvents or cosolutes, nonrigid compounds revealed a striking enhancement of fluorescence. The results indicate that these new or poorly known fluorochromes bind selectively to DNA-containing structures and that the minor groove from AT-rich DNA regions could represent the specific and highly fluorescent binding site.

New fluorescence reactions in DNA cytochemistry. 2. Microscopic and spectroscopic studies on fluorescent aluminum complexes.

Metal-dye complexes are widely applied in light microscopic techniques for chromatin staining (e.g., hematoxylin and carmine), but fluorescent complexes between phosphate-binding cations and suitable ligands have been little used. Preformed and postformed Al complexes with different anionic dyes induced strong and selective fluorescence reactions in nuclei from chicken blood smears, frozen sections, paraffin-embedded sections and Epon-embedded sections of mouse and rat tissues, mitotic chromosomes, meiotic chromosomes and kinetoplasts of Trypanosoma cruzi epimastigotes. The DNA-dependent fluorescence of these structures showed a very low fading rate. The emission colors were related to the ligand. The most suitable compounds for forming fluorescent Al chelates were 8-hydroxyquinoline, morin, nuclear fast red and purpurin. Staining with diluted carmine solutions and InCl3 mordanting, followed by 8-hydroxyquinoline, also induced chromatin fluorescence. After treating isolated mouse chromosomes with the preformed complex Al-nuclear fast red, x-ray microanalysis indicated a P:Al:dye binding ratio of about 40:15:1. The selectivity, stability and easy formation of these fluorescent Al complexes are obvious advantages for their use as new cytochemical probes in cytologic studies.

Observations on the contrasting reaction of some electron dense stains applied on epoxy-embedded tissue sections.

Epon sections from glutaraldehyde-fixed rat bone marrow were treated with aqueous solutions of the following electron contrasting agents: uranyl acetate, ruthenium red, potassium permanganate, potassium dichromate, stannous chloride, palladium (II) chloride, sodium molybdate, phosphomolybdic acid, molybdenum heteropolyblue, phosphotungstic acid, iron(II)-phenanthroline, aluminium-hematoxylin, mercurochrome, cuprolinic blue, and sirius light turquoise blue. At the ultrastructural level, a high degree of electron opacity was always observed in mast cell granules and the crystalline inclusion (internum) of eosinophil granules. The chromatin revealed a somewhat lower and variable contrasting reaction, while the matrix (externum) of eosinophil granules appeared with scarce or no contrast. This pattern of electron opacity showed no correlation with the type of agent used; therefore, it can be assumed that binding processes based on the own chemical reactivity of the compounds are rather of secondary importance. The differential epoxy resin embedding of cell structures and the variable access of aqueous reagents through the non-polar plastic could be the predominant factors which account for these contrasting reactions.

Chromatin fluorescence after carmine staining.

After staining with dilute solutions (0.1 mg/ml in distilled water) of commercial carmine, a strong reddish orange fluorescence was observed in nuclei from cell smears and frozen and paraffin tissue sections. Optimal exciting light was 436 nm (violet-blue) or 450-490 nm (blue). Compact chromatin from interphase nuclei, mitotic and meiotic chromosomes and the kinetoplast of Trypanosoma cruzi showed the highest fluorescence, while the basophilic cytoplasm appeared weakly fluorescent. No emission was observed in cartilage matrix, mast cell granules or goblet cell mucin. This selective method could be valuable in microscopic and cytochemical studies on chromatin because the carmine fluorescence is stable and preparations can be dehydrated and mounted permanently without changes in the fluorescence pattern.