Effect of the anode material, applied current and reactor configuration on the atenolol toxicity during an electrooxidation process.

Atenolol (ATL) is a beta-blocker pharmaceutical product which is excreted mainly unchanged and may represent a long-term risk for organisms present in the sea and in fresh water. Due to its low biodegradation rate, electrochemical advanced oxidation processes (EAOPs) can be used to remove this compound. In this work, ATL ecotoxicity was analyzed in the presence of sodium sulfate (Na2SO4), which is widely used as supporting electrolyte in EAOPs. Ecotoxicity values were expressed as the pollutant concentration that leads to a 50% inhibition of the root elongation of Lactuca sativa seeds in relation to the control (EC50(5 days)). The obtained values for ATL showed an EC50(5 days) of 1377 mg L-1 towards Lactuca sativa. When Na2SO4 was added, the toxicity of the sample increased but no synergy was detected between both compounds. With 2 g L-1 Na2SO4, ATL showed an EC50(5 days) of 972 mg L-1; and with 4 g L-1 Na2SO4 and higher concentrations, EC50 value for ATL was 0 mg L-1. Statistical tools were used to obtain the zones of the [ATL]-[Na2SO4] plane which are toxic towards Lactuca sativa. Solutions containing ATL and Na2SO4 were treated by electrooxidation. Two anode materials (a boron-doped diamond electrode and a microporous Sb-doped SnO2 ceramic one); three operation currents (0.4, 0.6 and 1 A); and two reactor configurations (one-compartment reactor and two-compartment reactor separated by a cation exchange membrane) were used. Lactuca sativa seeds and Vibrio fischeri bacterium tests were employed to evaluate the toxicity of the solutions before and after applying the electrooxidation process. In all the tests, the ecotoxicity of the treated sample increased. This fact is owing to the persulfate presence in the solution due to the sulfate electrochemical oxidation. Nevertheless, none of the final samples were toxic towards Vibrio fischeri because ecotoxicity values were lower than 10 TU; and, in the case of the one-compartment reactor, practically all of them were also non-toxic towards Lactuca sativa. The toxicity of the treated samples increased when using the two-compartment reactor in the presence of the BDD anode, and when the operation current was increased. This is attributed to the highest formation of persulfates. Amongst all the tests performed in this work, the lowest toxicity value (i.e., 3 TU) together with the complete mineralization and degradation degrees was achieved with the two-compartment reactor using the BDD anode and operating at 0.6 A.

Novel Sb-doped SnO2 ceramic anode coated with a photoactive BiPO4 layer for the photoelectrochemical degradation of an emerging pollutant.

In the present work, a study about the electrochemical and photoelectrochemical degradation of an emerging pollutant using an Sb-doped SnO2 anode coated with a photocatalytic layer of BiPO4 has been performed. The electrochemical characterization of the material was carried out by means of linear sweep voltammetry, light-pulsed chronoamperometry and electrochemical impedance spectroscopy. These studies confirmed that the material is photoactive at intermediate potential values (around 2.5 V), and that the charge transfer resistance decreases in the presence of light. A positive effect of the illuminated area on the degradation degree of norfloxacin was observed: at 15.50 mA cm-2, the degradation rate was 83.37% in the absence of light, 92.24% with an illuminated area of 5.7 cm2, and it increased up to 98.82% with an illuminated area of 11.4 cm2. The kinetics of the process were evaluated, and the by-products of the degradation were identified by ion chromatography and HPLC. In the case of the mineralization degree, the effect of light is less significant, especially at higher current densities. The specific energy consumption of the process was lower in the photoelectrochemical experiments as compared to the experiments in dark conditions. At intermediate current densities (15.50 mA cm-2) a decrease in energy consumption of 53% was achieved by illuminating the electrode.

Norfloxacin mineralization under light exposure using Sb-SnO2 ceramic anodes coated with BiFeO3 photocatalyst.

Advanced Oxidation Processes have been proven to be an efficient way to remove organic pollutants from wastewaters. In this work, a ceramic electrode of Sb-SnO2 (BCE) with a layer of the photocatalytic material BiFeO3 (BFO-BCE), has been characterized electrochemically and further tested for norfloxacin photo-electrooxidation in the presence and absence of light. The electrode photoactivity was highly enhanced thanks to the presence of BiFeO3, as confirmed by Linear Sweep Voltammetry, chronoamperometry and potentiometry, and Electrochemical Impedance Spectroscopy. Additionally, working in galvanostatic mode, a high mineralization of norfloxacin was achieved after 240 min, reaching 62% at 25 mA cm-2 under light conditions. This value is comparatively higher than the 40% achieved with the BCE. The oxidation byproducts were followed by ionic chromatography and HPLC analysis, which also allowed us to propose an oxidation pathway of the norfloxacin molecule. Finally, some indicators of the reactor performance such as the Mineralization Current Efficiency and the specific energy consumption were analyzed, revealing that lower current densities (8.3 mA cm-2) led to higher current efficiencies, and that light improved both the current efficiency and energy consumption.

Study of the chlorfenvinphos pesticide removal under different anodic materials and different reactor configuration.

The present manuscript focuses on the study of the electrochemical oxidation of the insecticide Chlorfenvinphos (CVP). The assays were carried out under galvanostatic conditions using boron-doped diamond (BDD) and low-cost tin dioxide doped with antimony (Sb-doped SnO2) as anodes. The influence of the operating variables, such as applied current density, presence or absence of a cation-exchange membrane and concentration of supporting electrolyte, was discussed. The results revealed that the higher applied current density the higher degradation and mineralization of the insecticide for both anodes. The presence of the membrane and the highest concentration of Na2SO4 studied (0.1 M) as a supporting electrolyte benefited the oxidation process of CVP using the BDD electrode, while with the ceramic anode the elimination of CVP was lower under these experimental conditions. Although the BDD electrode showed the best performance, ceramic anodes appear as an interesting alternative as they were able to degrade CVP completely for the highest applied current density values. Toxicity tests revealed that the initial solution of CVP was more toxic than the samples treated with the ceramic electrode, while using the BDD electrode the toxicity of the sample increased.

A U-Net based framework to quantify glomerulosclerosis in digitized PAS and H&E stained human tissues.

Reliable counting of glomeruli and evaluation of glomerulosclerosis in renal specimens are essential steps to assess morphological changes in kidney and identify individuals requiring treatment. Because microscopic identification of sclerosed glomeruli performed under the microscope is labor intensive, we developed a deep learning (DL) approach to identify and classify glomeruli as normal or sclerosed in digital whole slide images (WSIs). The segmentation and classification of glomeruli was performed by the U-Net model. Subsequently, glomerular classifications were refined based on glomerular histomorphometry. The U-Net model was trained using patches from Periodic Acid-Schiff (PAS) stained WSIs (n=31) from the AIDPATH - a multi-center dataset, and then tested on an independent set of WSIs (n=20) including PAS (n=6), and hematoxylin and eosin (H&E) stained WSIs (n=14) from four other institutions. The training and test WSIs were obtained from formalin fixed and paraffin embedded blocks with of human kidney specimens each presenting various proportions of normal and sclerosed glomeruli. In the PAS stained WSIs, normal and sclerosed glomeruli were respectively classified with the F1-score of 97.5% and 68.8%. In the H&E stained WSIs, the F1-scores of 90.8% and 78.1% were achieved. Regardless the tissue staining, the glomeruli in the test WSIs were classified with the F1-score of 94.5% (n=923, normal) and 76.8% for (n=261, sclerosed). These results demonstrate for the first time that a framework based on the U-Net model trained with glomerular patches from PAS stained WSIs can reliably segment and classify normal and sclerosed glomeruli in PAS and also H&E stained WSIs. Our approach yielded higher accuracy of glomerular classifications than some of the recently published methods. Additionally, our test set of images with ground truth is publicly available.

Comparison between an electrochemical reactor with and without membrane for the nor oxidation using novel ceramic electrodes.

The electrochemical oxidation of the antibiotic Norfloxacin (NOR) in chloride media on different anodic materials was studied at two different electrochemical reactors. The results were compared with those obtained in sulphate media. The anodes under study were a commercial boron-doped diamond (BBD) and two different ceramic electrodes based on tin oxide doped with antimony oxide in the presence (CuO) and absence (BCE) of copper oxide as sintering aid. The reactors employed were a one-compartment reactor (OCR) and a two-compartment one with a membrane separating both electrodes (EMR). The use of the membrane clearly enhanced both NOR degradation and TOC mineralization for all the anodic materials studied since some parallel reactions were avoided. Additionally, two different pathways for NOR oxidation were observed as a function of the reactor employed. The EMR also favoured the ionic by-products generation and the electrolyte dechlorination. NO3- increased with the oxidation power of the anode employed and it was also enhanced by the EMR use. Chloride media favours ceramic electrodes performance independently of the reactor employed as they did not generate an excess of oxidants as BDD did. The BCE electrode is an interesting alternative to BDD since although its oxidative power was lower, it presented similar current efficiency with lower energy consumption.

Effect of the CuO addition on a Sb-doped SnO2 ceramic electrode applied to the removal of Norfloxacin in chloride media by electro-oxidation.

Norfloxacin is employed as in veterinary and human medicine against gram-positive and gram-negative bacteria. Due to the ineffective treatment at the wastewater treatment plants it becomes an emergent pollutant. Electro-oxidation appears as an alternative to its effective mineralization. This work compares Norfloxacin electro-oxidation on different anodic materials: two ceramic electrodes (both based on SnO2 + Sb2O3 with and without CuO, named as CuO and BCE, respectively) and a boron doped diamond (BDD). First, the anodes were characterized by cyclic voltammetry, revealing that NOR direct oxidation occurred at 1.30 V vs. Ag/AgCl. The higher the scan rate the higher both the current density and the anodic potential of the peak. This behavior was analyzed using the Randles-Sevcik equation to calculate the Norfloxacin diffusion coefficient in aqueous media, giving a value of D = 7.80 × 10-6 cm2 s-1 at 25 °C), which is close to the predicted value obtained using the Wilke-Chang correlation. The electrolysis experiments showed that both NOR and TOC decay increased with the applied current density, presenting a pseudo-first order kinetic. All the anodes tested achieved more than 90% NOR degradation at each current density. The CuO is not a good alternative to BCE because although it acts as a catalyst during the first use, it is lost from the anode surface in the subsequent uses. According to their oxidizing power, the anodes employed are ordered as follows: BDD > BCE > CuO.

Analysis of norfloxacin ecotoxicity and the relation with its degradation by means of electrochemical oxidation using different anodes.

In this work, ecotoxicological bioassays based on Lactuca sativa seeds and bioluminescent bacterium (Vibrio fischeri) have been carried out in order to quantify the toxicity of Norfloxacin (NOR) and sodium sulfate solutions, before and after treating them using electrochemical advanced oxidation. The effect of some process variables (anode material, reactor configuration and applied current) on the toxicity evolution of the treated solution has been studied. A NOR solution shows an EC50(5 days) of 336 mg L-1towards Lactuca sativa. This threshold NOR concentration decreases with sodium sulfate concentration, in solutions that contain simultaneously Norfloxacin and sodium sulfate. In every case considered in this work, the electrochemical advanced oxidation process increased the toxicity (towards both Lactuca sativa and Vibrio fischeri) of the solution. This toxicity increase is mainly due to the persulfate formation during the electrochemical treatment. From a final solution toxicity point of view, the best results were obtained using a BDD anode in a divided reactor applying the lowest current intensity.

Trypanosoma cruzi: A kinetoplast-associated protein of the photolyase/cryptochrome family.

A photolyase-like protein gene found in the Trypanosoma cruzi genome database was cloned and expressed in Escherichia coli resulting in the formation of inclusion bodies. Antibodies against this protein were used to determine expression of the protein in the different forms of the parasite. It was visualized in the epimastigote form but not in amastigote or trypomastigote forms obtained from culture in Vero cells. In epimastigotes, this protein is located at the level of the mitochondrion associated to both sides of the kinetoplast. Sequence analyses indicated that this protein, as well as other photolyases from Leishmania spp. and Trypanosoma brucei are related to single-stranded photolyases or cryptochromes DASH.

Effect of inflammatory stimuli on the silver staining pattern of the rat carotid endothelium.

Silver nitrate stains the intercellular junctions of the endothelium and other cytoplasmic or membrane components. Two protocols are described for the silver staining of rat carotid endothelium that exclude the use of pressurized fixatives and simplify the technique previously described for rat aorta. The entire surface of the carotid endothelium was examined and several parameters (stigmata, granularity, clustering of anionic sites, transversal lines, weakening of silver lines and leukocyte adhesion) were evaluated. We studied the pattern of silver staining in two situations: (1) endothelial activation and (2) neurogenic inflammation. Endothelial activation was produced by the intravenous administration of a proinflammatory albumin or polyinosinic acid. Both products cause a marked increase in leukocyte adhesion concomitant with a decrease in argyrophilia and a weakness or loss of silver lines. Neurogenic inflammation, which is mediated by substances released from sensory nerves, was induced by the intravenous administration of substance P or capsaicin. Both stimuli produced an increase in argyrophilia and weakness or loss of silver lines. Substance P caused a clustering of anionic sites, whereas this phenomenon was more discrete with capsaicin. Nearly 80% of all examined rats (controls and inflammatory stimuli treated) showed endothelial membrane disruptions formed by clusters of cells often in the shape of streaks aligned with the long axis of the vessel. The detection of these discontinuities is important, as loss of endothelial integrity is central in the initiation of pathological events.

Proinflammatory activity on rat carotid endothelium of albumins obtained by different procedures.

To investigate whether the proinflammatory activity of intravenously administered albumins is related to their preparation method, we compared bovine albumins obtained by alcohol precipitation and by heat shock. This last procedure produced a notable increase in monocyte adhesion/mm2 in rat carotid endothelium (48 +/- 31, n = 32 rats, vs 7 +/- 6, n = 24 rats, mean +/- S.D., P < 0.001). We found no changes in fluorescence emission in these two groups, and no correlation was observed between leukocyte adhesion and the contents of free fatty acids, free -SH groups, fructosamine, dimers and polymers. The inhibition profile of the proinflammatory activity by polyanionic molecules (fucoidan and dextran sulfate 500 kD) was compatible with the recognition of heat-shocked albumins by scavenger receptors. Thiol disulfide exchange in albumins was produced by ageing with cysteine and by incubation with protein disulfide isomerase. Albumins thus modified increased the adhesion of leukocytes to the endothelium, which indicates that some isomeric forms of albumin generated by thiol disulfide exchange may acquire proinflammatory behavior.

Effect of hyperlipidemic diets, proinflammatory agents and plasma expanders on leukocyte adhesion to the endothelium of aorta and carotid artery of rats.

A comparative study of leukocyte adhesion to the endothelium of the thoracic aorta and left carotid artery in rats has been performed after administration of two hyperlipidemic diets for 15 days, proinflammatory agents (thrombin, lipopolysaccharide and zymosan activated serum) and plasma expanders [dextran, polyvinylpyrrolidone (PVP), rat albumin and several bovine albumins from different sources]. Leukocytes adhered to the endothelium were demonstrated in surface preparations by esterase activity. Activation of circulating leukocytes was measured by nitroblue tetrazolium reduction and luminol enhanced chemiluminescence. Both hyperlipidemic diets produced, in all rats, more leukocyte adhesion in the aorta than in the carotid artery. All proinflammatory agents produced at 1 h, increases in leukocyte adhesion--which in all rats were greater in the carotid artery than in the aorta--and leukocyte activation, which was higher at 3 h than at 1 h. Dextran, PVP, bovine albumins 103700 and A-4503 at 18 h produced slight increases in leukocyte adhesion in the aorta but not in the carotid artery. Rat albumin and bovine albumin A-7906 determined an intense leukocyte adhesion at 18 h which was not preferential to either vessel. Adhesion produced by A-7906 was maximal at 12 h and partially inhibited by dexamethasone. This last albumin produced leukocyte activation at 3 h and was sequestered 5 min after administration, reaching normal values at 1 h. Albumins 103700 and A-4503 neither activated leukocytes nor were sequestered after administration.

Phagocytosis of intravenously administered particles by leukocytes adhered to the aortic endothelium of the rat.

Phagocytosis has been used to characterize on a functional basis leukocytes adhered to the aortic endothelium of the rat. After intravenous administration of particles, phagocytosis was observed microscopically in esterase-positive leukocytes adhered to the endothelium in whole mounts of aorta. Polybead blue and red, 0.5 and 1 microns particle size, were inadequate because they were insufficiently colored to be identified individually at 400 x. Fluoresbrite YG 0.25 and 0.50 micron at doses of 0.2 and 2 x 0.3 ml/100 g, respectively, produced endothelial lesions. The same occurred with Monastral blue B (MbB) at 0.3 ml/100 g, red iron at 2 x 16 mg/100 g and India ink at different concentrations depending on the supplier. At lower particle doses, lesions were not found. Deferoxamine mesylate 1.5 mg/100 g intravenous and allopurinol 5 mg/100 g intraperitoneal administered before the particles diminished the number and intensity of lesions. In none of the cases studied was the percentage of phagocytic cells greater than 50%. Clearance curves of MbB and Fluoresbrite indicated rapid disappearance of particles from the blood. Results indicate that administration of particulate suspensions is not a good method for characterizing the phagocytic leukocytes adhering to the aortic endothelium because low doses produce rapid clearance of particles, thus impeding sufficient leukocyte loading, and higher doses produce endothelial lesions that often impair reliable counting of the adhering leukocytes.

Body-image distortion among male and female American and Costa Rican students and female Japanese students.

395 American male and female high school and college students, 286 Costa Rican male and female high school students, and 127 Japanese female college students were tested for body-image distortion using a computer-based body-image distortion task. A reliable negative relationship between stature and body-image distortion was observed. No reliable differences in body-image distortion were observed between different sex or cultural groups.

Body-image dissatisfaction among American male and female college students: a computer-based graphical approach.

College students (56 women and 43 men) attending state colleges in the southwestern United States were tested for body-image dissatisfaction using a computer-based graphical body-image task. A reliable relationship between desired stature and desired body-image was observed for the women. Women of large stature showed a greater discrepancy between verbally reported desired stature and redrawn images of desired stature than women of average or smaller than average stature. No reliable discrepancy between desired body-image and verbally reported desired stature was shown by the men.

Effect of spontaneous pathology and thrombin on leukocyte adhesion to rat aortic endothelium.

Leukocyte adhesion and other injury parameters have been studied in the aortic endothelium of Sprague-Dawley rats in two situations: (1) spontaneous pathology in conventional rats with antibodies to Mycoplasma pulmonis and/or Kilham or Sendai viruses, and (2) intravascular coagulation by thrombin administration in SPF rats. Adhesion (esterase (+) leukocytes/mm2) in SPF rats was 8 +/- 5 (n = 12). Adhesion in 38% of the conventional rats was 54 +/- 27 (n = 8), half of them being non-analyzed and the rest having antibodies to M. pulmonis and/or Kilham rat virus. In 19 rats with antibodies to M. pulmonis and/or Kilham or Sendai viruses, AgNO3 and hematoxylin staining of the aortic endothelium showed an increase in leukocyte adhesion, and the presence of argyrophilic cells, stigmata and granularity--severe endothelial lesions being observed in some cases. Adhesion in rats after 0.25, 1, 3 and 6 h of thrombin administration (30 units/100 g) was not different from controls. Adhesion after 24 h was 108 +/- 53 (n = 10) and 60 +/- 59 (n = 10), and 22 +/- 20 (n = 10) in rats treated with thrombin plus heparin or hirudin, respectively. Thrombin produced endothelial lesions at all times studied, and these included membrane blebs, platelet and erythrocyte adhesion and alterations in the pattern of endothelial esterase activity.

Technical considerations in evaluating the endothelial integrity of rat aortic preparations with silver staining.

Assessment of endothelial integrity is an obligatory step in many pharmacological studies. Integrity of endothelium is affected by manipulations performed during the removal and cleaning of the vessel and by some of the silver-staining techniques utilized for demonstrating interendothelial junctions. When aortas were cleaned of periadventitial tissue in cold Tris-saline (once separated from the animal) by untrained personnel, only 45% of the endothelium was preserved. When cleaning was performed in situ by trained personnel while flushing with cold Krebs-Ringer-6% albumin, over 95% was left intact. AgNO3-staining performed before fixation produced a 50% loss of endothelium when using NH4Br and (NH4)2S as developers. AgNO3-staining performed after fixation produced over 95% recuperation of endothelium when 2% glutaraldehyde, 150 mM NaCl, 40 mM phosphate buffer, pH 7.4, were utilized as initial fixative, NH4Br and (NH4)2S being equally effective as developers. Chloride ions were necessary to intensify silver lines. Several patterns of deendothelization were produced by mechanical and chemical injury with saponin, NH4Br and (NH4)2S. In all cases, hematoxylin staining was employed as an auxiliary technique to interpret images of injured endothelium. Presence of albumin protected the endothelium from mechanical damage.

Technical considerations in evaluating the adhesion of leukocytes to aortic endothelium of the rat.

Comments on techniques for characterizing leukocytes adhered to the aortic endothelium of the rat are given. Alpha-naphthyl acetate esterase positive leukocytes were studied by optical microscopy of en face intima-media preparations. Results indicate 1) 1% paraformaldehyde-2% glutaraldehyde is a better fixative than formalin-calcium or 4% paraformaldehyde with or without 1.5 mM CaCl2; the latter produces distortion of leukocytes, endothelial desquamation and enzymate inhibition, 2) washing the aorta with phosphate-buffered saline for 90 sec prior to fixation-perfusion produces a notable decrease in the number of leukocytes adhered, 3) diazotized parasaniline is better than fast blue RR salt as coupling agent in the esterase reaction, and 4) counterstaining with 1% methyl green for 1 min, before or after the esterase reaction, is not adequate because of limited contrast and the heavy staining of smooth muscle. Counterstaining with Gill's hematoxylin No. 3 for 90 sec is adequate only when done before the esterase reaction. Inhibition of endothelial esterase activity by hematoxylin decreases background, favors contrast of adhered leukocytes and makes it possible to observe nucleus-cytoplasm relations.

Adhesion of leukocytes to the aortic endothelium of conventional, specific pathogen free (SPF) and hypercholesterolemic SPF rats.

Adhesion of leukocytes to the aortic endothelium was studied in specific pathogen-free (SPF) and conventional rats and in SPF rats with diet-induced hypercholesterolemia. Nonspecific esterase activity with alpha-naphthyl acetate as substrate was used to characterize the adhered cells. Phagocytic activity was determined by injecting i.v. 0.1-0.4 ml/100 g doses of Monastral blue B (MbB). Adhesion in SPF rats was 8 +/- 4 esterase (+) cells/mm2. Adhesion in conventional rats was of the same order except in 2 cases with antibodies to Mycoplasma pulmonis and Kilham rat virus, where adhesion was 44 and 68 esterase (+) cells/mm2, respectively. For all MbB doses studied, phagocytic activity arose in a percentage of the adherent cells, ranging from 5 to 85%. Rats fed the hyperlipidic diet for 15 days developed severe hypercholesterolemia and adhesion was drastically increased to 200-700 esterase (+) cells/mm2. Results indicate that: (1) spontaneous pathology in rats may produce an increased adhesion of leukocytes to the endothelium, and (2) phagocytic activity is only expressed in a fraction of the esterase (+) cells adhered to the endothelium.