Copper sulfate and arginine supplements on performance, thyroid hormones and blood biochemistry of broilers fed with canola meal-based diets / Avaliação da suplementação com sulfato de cobre e arginina sobre o desempenho, hormônios tireoidianos e bioquímica sanguínea de frangos alimentados com dietas à base de canola

The present evaluated the effects of copper sulfate solution (CSS) and arginine powder (Arg) supplements on performance, thyroid hormones and blood biochemistry of broiler chickens fed with canola meal (CM)-based diets. The experimental design was completely randomized with a 3 × 3 factorial and 9 treatments, corresponding to 3 levels of CSS (0, 125 and 250 mg/kg) and 3 levels of Arg (0, 0.1 and 0.2%) (n = 45 per treatment). Feeds were offered ad libitum for 21 days, from 22 to 42 days of age. Feed efficiency was significantly affected by the dietary addition of 250 mg/kg CSS and 0.2% Arg, and by the CSS × Arg interaction. CM supplemented with CSS improved the thyroid gland status and increased the plasma levels of triiodothyronine and thyroxine. Birds fed diets supplemented with 0.2% Arg had lower blood glucose level than the other treatments. The addition of 250 mg/kg CSS and 0.2% Arg reduced the stress caused by the rapid growth of broilers, also increasing the overall bird welfare.

O objetivo do presente trabalho foi avaliar os efeitos da suplementação com solução de sulfato de cobre (SSC) e arginina em pó (Arg) sobre o desempenho, hormônios tireoidianos e bioquímica sanguínea de frangos de corte alimentados com dietas à base de canola DC. O desenho experimental foi completamente casualizado com fatorial 3 × 3 e nove tratamentos correspondentes a três níveis de inclusão de SSC (0, 125 e 250 mg/kg) e três níveis de Arg (0, 0,1 e 0,2%) (n = 45 para cada tratamento). As rações foram oferecidas ad libitum por 21 dias, de 22 até 42 dias de idade. A eficiência alimentar foi significativamente afetada pela adição de 250 mg/kg de SSC e 0,2% de Arg, assim como pela interação SSC × Arg. A suplementação da DC com SSC melhorou os parâmetros da glândula tireoide e aumentou os níveis plasmáticos de triiodotironina e tiroxina. As aves alimentadas com dietas suplementadas com 0,2% de Arg apresentaram menor nível de glicose sanguínea do que as dos demais tratamentos. A adição de 250 mg/kg de SSC e 0,2% de Arg reduz o estresse causado pelo rápido crescimento dos frangos, além de melhorar as condições gerais de bem estar das aves.

Efficacy of copper sulphate on candida albicans on heat-polymerized acrylic resin / Eficacia del sulfato de cobre sobre candida albicans en resinas acrílicas de termocurado

The ageing of population is increasing, and a great percentage of these patients wear removable prostheses, and can suffer denture stomatitis, a condition that has been associated with candidiasis. Aims: To evaluate in vitro the effectiveness of Copper Sulfate against Candida albicans in samples of heat-polymerized acrylic resin, compared to nystatin, sodium hypochlorite and chlorhexidine. Materials and Methods: Initially, the minimum inhibitory concentration (MIC) of copper sulfate for Candida albicans was determined by microdilution. Then, 54 resin samples were divided into 6 treatment groups corresponding to Nystatin 100.000 UI, Sodium Hypochlorite 0.5%, chlorhexidine 0.12%, Copper Sulfate 4.7µg/ml, Copper Sulfate 9.4µg/ml and physiological saline solution, in which samples were submerged for 6 hours. Resin samples were then washed and cultured on solid media at 37°C for 72 hours. The number of colony-forming units was determined using a Quebec colony counter. The statistical analysis was performed using the Kruskal-Wallis test and the Mann-Whitney U test. Results: Copper sulfate at a concentration of 9.4µg/ml presented a similar effectiveness as the other control products regarding the reduction in the number of colonies of Candida albicans post-treatment. Conclusion: The effectiveness of copper sulfate against Candida albicans on acrylic resin was similar to that of nystatin, sodium hypochlorite and chlorhexidine.

En las últimas décadas se ha observado un aumento de la población de adultos mayores, de los cuales un gran porcentaje es portador de prótesis removible, y dos tercios pueden sufrir estomatitis subprotésica, enfermedad que es asociada a infecciones como candidiasis. Objetivo: Evaluar la efectividad antimicótica in vitro del sulfato de cobre en placas de resinas acrílicas de termocurado inoculadas con Candida albicans, frente a Nistatina, Hipoclorito de Sodio y Clorhexidina. Material y Métodos: Inicialmente, y mediante microdilución del sulfato de cobre, se determinó la concentración mínima inhibitoria (CMI) para Candida albicans. En la fase experimental, 54 muestras de resina se dividieron en 6 grupos correspondientes a Nistatina 100.000 UI, Hipoclorito 0.5%, Clorhexidina 0.12%, Sulfato de Cu 4.7µg/ml, Sufato de Cu 9.4 µg/ml y suero fisiológico. Las muestras fueron sumergidas en estos agentes por 6 horas, para posteriormente ser lavadas y cultivada en medios solidos a 37°C por 72 horas. Luego se realizó el conteo de unidades formadoras de colonias mediante contador tipo Quebec. El análisis estadístico se realizó mediante la prueba de Kruskal-Wallis y la prueba U de Mann-Whitney. Resultado: El sulfato de cobre a una concentración de 9.4µg/ ml presentó una efectividad similar a los otros productos, en la reducción de colonias de Candida albicans. Conclusión: La efectividad del sulfato de cobre contra Candida albicans fue semejante a la de Nistatina, Hipoclorito y Clorhexidina.

Improvement of anther cultures conditions using the Taguchi method in three cereal crops

Background: Plant tissue cultures have the potential to reprogram the development of microspores from normal gametophytic to sporophytic pathway resulting in the formation of androgenic embryos. The efficiency of this process depends on the genotype, media composition and external conditions. However, this process frequently results in the regeneration of albino instead of green plants. Successful regeneration of green plants is affected by the concentration of copper sulfate (CuSO4) and silver nitrate (AgNO3) and the length of induction step. In this study, we aimed at concurrent optimization of these three factors in barley (Hordeum vulgare L.), wheat (Triticum aestivum L.), and triticale (x Triticosecale spp. Wittmack ex A. Camus 1927) using the Taguchi method. We evaluated uniform donor plants under varying experimental conditions of in vitro anther culture using the Taguchi approach, and verified the optimized conditions. Results: Optimization of the regeneration conditions resulted in an increase in the number of green regenerants compared with the control. Statistic Taguchi method for optimization of the in vitro tissue culture plant regeneration via anther cultures allowed reduction of the number of experimental designs from 27 needed if full factorial analysis is used to 9. With the increase in the number of green regenerants, the number of spontaneous doubled haploids decreased. Moreover, in barley and triticale, the number of albino regenerants was reduced. Conclusion: The statistic Taguchi approach could be successfully used for various factors (here components of induction media, time of incubation on induction media) at a one time, that may impact on cereals anther cultures to improve the regeneration efficiency

Efecto del sulfato de cobre en el daño pulmonar inducido por la exposición crónica intermitente a ozono / Effect of copper sulphate on the lung damage induced by chronic intermittent exposure to ozone

Background: Ozone exposure could increase lung damage induced by airborne particulate matter. Particulate matter lung toxicity has been attributed to its metallic content. Aim: To evaluate the acute effect of intratracheal administration of copper sulfate (CuSO4) on rat lungs previously damaged by a chronic intermittent ozone exposure. Material and Methods: Two-months-old male Sprague-Dawley rats were exposed to 0.5 ppm ozone four h per day, five days a week, during two months. CuSO4 was intratracheally instilled 20 h after ozone exposure. Controls breathed filtered air or were instilled with 0.9% NaCl or with CuSO4 or were only exposed to ozone. We evaluated lung histopathology. F2 isoprostanes were determined in plasma. Cell count, total proteins, γ glutamyl-transpeptidase (GGT) and alkaline phosphatases (AP) were determined in bronchoalveolar lavage fluid (BALF). Results: Ozone increased total cell count, macrophages, proteins and AP in BALF (p < 0.05), and induced pulmonary neutrophil inflammation. CuSO4 plus air increased plasma F2 isoprostane levels and total cell count, neutrophils and proteins in BALF (p < 0.05). Histopathology showed foamy macrophages. Ozone plus CuSO4 exposed animals showed a neutrophil inflammatory lung response and an increase in total cell count, proteins, GGT and AP in BALF (p < 0.05). Foamy and pigmented alveolar macrophages were detected in all lungs of these animals (p < 0.001). Conclusions: Intratracheal instillation of a single dose of CuSO4 in rats previously subjected to a chronic and intermittent exposure to ozone induces a neutrophil pulmonary inflammatory response and cytoplasmic damage in macrophages.

Physiological Regulation of an Alkaline-Resistant Laccase Produced by Perenniporia tephropora and Efficiency in Biotreatment of Pulp Mill Effluent

Regulation of alkaline-resistant laccase from Perenniporia tephropora KU-Alk4 was proved to be controlled by several factors. One important factor was the initial pH, which drove the fungus to produce different kinds of ligninolytic enzymes. P. tephropora KU-Alk4 could grow at pH 4.5, 7.0, and 8.0. The fungus produced laccase and MnP at pH 7.0, but only laccase at pH 8.0. The specific activity of laccase in the pH 8.0 culture was higher than that in the pH 7.0 culture. At pH 8.0, glucose was the best carbon source for laccase production but growth was better with lactose. Low concentrations of glucose at 0.1% to 1.0% enhanced laccase production, while concentrations over 1% gave contradictory results. Veratryl alcohol induced the production of laccase. A trace concentration of copper ions was required for laccase production. Biomass increased with an increasing rate of aeration of shaking flasks from 100 to 140 rpm; however, shaking at over 120 rpm decreased laccase quantity. Highest amount of laccase produced by KU-Alk4, 360 U/mL, was at pH 8.0 with 1% glucose and 0.2 mM copper sulfate, unshaken for the first 3 days, followed by addition of 0.85 mM veratryl alcohol and shaking at 120 rpm. The crude enzyme was significantly stable in alkaline pH 8.0~10.0 for 24 hr. After treating the pulp mill effluent with the KU-Alk4 system for 3 days, pH decreased from 9.6 to 6.8, with reduction of color and chemical oxygen demand at 83.2% and 81%, respectively. Laccase was detectable during the biotreatment process.

Ex vivo effect of copper sulfate on Enterococcus faecalis in root canals / Efecto ex vivo del sulfato de cobre sobre Enterococcus faecalis en canales radiculares

The incomplete disinfection of root canal system has been reported as the main cause of post-treatment disease, due to the persistence of bacteria. For over thirty years Enterococcus faecalis has been considered the most common bacterial species isolated from persistent root canal infections, resisting antibacterial agents, such as chlorhexidine and calcium hydroxide. Several studies have indicated that copper has optimal disinfecting capacities in a hospital environment. Aimed to know the ex vivo effect of copper sulfate (CuSO4) on E. faecalis. Thirty-six extracted human tooth root canals were inoculated with E. faecalis ATCC 29212. These root canals had undergone endondontic procedures with a rotatory system. The effect of CuSO4 was determined by plate count method of E. faecalis obtained from one sample of each tooth under three incubation times (4th, 7th and 10th day). The canals medicated with CuSO4, the bacterial count decreased 6 log after 4 days and remained as such without statistically significant change until the tenth day. It is an indisputable fact of its antibacterial action. The bacterial persistence may be due to the ability of E. faecalis to remain viable in root canals up to 12 months without additional nutrients. These preliminary results couldbe used for further scientific work assessing the potential for the use of cooper in dentistry.

La desinfección incompleta del sistema de canales radiculares ha sido reportada como la principal causa de infección post tratamiento, debido a la persistencia bacteriana. Por más de 30 años, Enterococcus faecalis se ha considerado una de las especies bacterianas más comunes aisladas de infecciones persistentes del canal radicular, resistiendo a agentes antibacterianos, como clorhexidina e hidróxido de calcio. Varios estudios han indicado que el cobre tiene una capacidad de desinfección óptima en un entorno hospitalario. Con el objetivo de conocer el efecto ex vivo de sulfato de cobre (CuSO4) sobre E. faecalis. Treinta y seis canales radiculares de dientes humanos extraídos fueron inoculados con E. faecalis ATCC 29212. Estos canales radiculares fueron previamente sometidos a instrumentación con un sistema de rotación. El efecto de CuSO4 se determinó por el método de recuento en placa de E. faecalis a partir de una muestra de cada diente en tres tiempos de incubación (4, 7 y 10 días). En los canales medicados con CuSO4, el recuento de bacterias se redujo 6 log después de 4 días y se mantuvo como tal sin cambio estadísticamente significativo hasta el día 10. Esto demuestra su poder antibacteriano. La persistencia bacteriana pudo ser deberse a la capacidad de E. faecalis para seguir siendo viable en canales hasta 12 meses sin nutrientes adicionales. Estos resultados preliminares podrían ser utilizados para otros trabajos científicos que evaluen el potencial uso de cobre en odontología.

2000 CDC or 2007 WHO – What is the most sensitive anthropometric reference for determination of overweight and cardio-metabolic risk in children aged 6–10 years?

Objective To compare the two anthropometric standards for screening of overweight and cardio-metabolic risk in 6–10-year-old children.Subjects and methods This cross-sectional study included 175 subjects attending the Referral Center for the Treatment of Children and Adolescents in Campos, Rio de Janeiro. They were classified according to CDC and WHO BMI z scores as normal-weight (z-score > –1 and < 1), overweight (z-score ≥ 1 and < 2) or obese (z-score ≥ 2). Sensitivities and specificities in predicting systolic (SBP), diastolic (DBP) blood pressure and homeostatic model assessment insulin resistance index (HOMA-IR) alterations were calculated.Results There was a major difference in 11 children who rated overweight by the CDC but were reclassified as obese by the WHO. Their mean z-scores for SBP (1.71 ± 1.54), DBP (2.64 ± 1.83) and HOMA-IR (1.84 ± 0.98) were higher than those classified as overweight by both references (SBP = 0.49 ± 1.34, p < 0.023, DBP = 1.45 ± 0.97, p < 0.04 and HOMA = 1.24 ± 0.67, p < 0.04), but were similar to those classified as obese by both criteria (SBP = 1.25 ± 2.04, p = 0.60, DBP = 1.94 ± 1.19, p = 0.50 and HOMA = 2.09 ± 1.12, p = 0.76).Conclusion the 2007 WHO reference was the most sensitive in screening for overweight and alterations in blood pressure and HOMA-IR in 6–10-year-old children. Arch Endocrinol Metab. 2015;59(3):220-5.

Acute Respiratory Failure due to Fatal Acute Copper Sulfate Poisoning: A Case Report

Copper sulfate is a copper compound used widely in the chemical and agriculture industries. Most intoxication occurs in developing countries of Southeast Asia particularly India, but rarely occurs in Western countries. The early symptoms of intoxication are nausea, vomiting, diarrhea, and abdominal cramps, and the most distinguishable clue is bluish vomiting. The clinical signs of copper sulfate intoxication can vary according to the amount ingested. A 75-year old man came to our emergency room because he had taken approximately 250 ml copper sulfate per oral. His Glasgow Coma Scale (GCS) score was 14 and vital signs were blood pressure 173/111 mmHg, pulse rate 24 bpm, respiration rate 24 bpm, and body temperature 36.1degrees.... Arterial blood gas analysis (ABGa) showed mild hypoxemia and just improved after 2 L/min oxygen supply via nasal cannula. Other laboratory tests and chest CT scan showed no clinical significance. Three hours later, the patient's mental status showed sudden deterioration (GCS 11), and ABGa showed hypercarbia. He was arrested and his spontaneous circulation returned after 8 minutes CPR. However, 22 minutes later, he was arrested again and returned after 3 minutes CPR. The family did not want additional resuscitation, so that he died 5 hours after ED visit. In my knowledge, early deaths are the consequence of shock, while late mortality is related to renal and hepatic failure. However, as this case shows, consideration of early definite airway preservation is reasonable in a case of supposed copper sulfate intoxication, because the patients can show rapid deterioration even when serious clinical manifestation are not presented initially.

Utilización de sulfato de cobre para la elaboración de un jabón líquido antiséptico / Use of cooper sulphate for production of antiseptic liquid soap

INTRODUCCIÓN: las manos poseen un papel importante en la transmisión de infecciones en instituciones de salud, entornos industriales como la industria alimentaria y en toda la comunidad, por lo que su higiene no puede pasar desapercibida. Los jabones líquidos son actualmente una herramienta fundamental para la limpieza de las manos y así también evitar la transmisión de agentes patógenos. OBJETIVO: evaluar la efectividad del sulfato de cobre como agente antiséptico en un jabón líquido y caracterizar el producto mediante controles de calidad. MÉTODOS: se evaluó la capacidad antiséptica del jabón líquido mediante el método de dilución en agar y la disminución del crecimiento bacteriano a través de una variación del recuento por microgota, a tiempos 1, 5 y 10 min. Posteriormente se realizaron controles fisicoquímicos, microbiológicos, prueba de parche y estudios de estabilidad en estantería y acelerada durante 6 meses. RESULTADOS: las cepas Staphylococcus aureus, Staphyloccus epidermidis, Escherichia coli, Pseudomona aeruginosay Klebsiella pneumoniae fueron sensibles a una concentración de 0,5 por ciento de sulfato de cobre. El crecimiento bacteriano en las cepas S. aureus y S. epidermidis mostró una reducción considerable en los tiempos 5 y 10 min a concentraciones de 0,3 y 0,5 por ciento de sulfato de cobre. El producto elaborado cumplió con los criterios de calidad establecidos, el cual mantuvo su estabilidad durante el tiempo de estudio. CONCLUSIONES: es posible elaborar un jabón líquido en base a sulfato de cobre como agente antiséptico, al ser eficaz frente a las cepas estudiadas(AU)

INTRODUCTION: the hands play a significant role in the transmission of infections at health institutions, industrial environments such as the food industry and at the community; therefore, hand hygiene cannot be ignored. Liquid soaps are now a fundamental tool for hand cleaning and prevention of pathogen transmission. OBJECTIVE: to evaluate the effectiveness of copper sulfate as an antiseptic agent in a liquid soap and to characterize the product by using quality controls. METHODS: aliquid soap was developed, in which its antiseptic capacity was evaluated through the agar dilution method and the reduction of bacterial growth by using a variant of microdrop counting at 1, 5 and 10 minutes. Then physicochemical and microbiological controls were performed, as well as patch test and shelf life and accelerated stability studies for six months. RESULTS: Staphylococcus aureus, Staphyloccus epidermidis, Escherichia coli, Pseudomona aeruginosa and Klebsiella pneumoniae were susceptible to 0.5 percent copper sulfate concentration. Bacterial growth showed considerable reduction in S. aureus and S. epidermidis strains at 5 and 10 minutes at copper sulphate concentrations of 0.3 and 0.5 percent. The finished product met the quality criteria for this type of product, maintaining its stability during the study period. CONCLUSIONS: it is possible to produce a copper sulfate-based liquid soap as an antiseptic since it was effective to control the studied strains(AU)

Chloroplast ultra structure, photosynthesis and enzyme activities in regenerated plants of Stevia rebaudiana (Bert.) Bertoni as influenced by copper sulphate in the medium.

Stevia rebaudiana (Bert.) Bertoni is an important medicinal plant used as noncaloric commercial sweetener. Plants regenerated with higher levels of copper sulphate in the medium exhibited enhanced activity of peroxidase and polyphenoloxidase (PPO) enzymes. Transmission electron microscopy (TEM) revealed increase in size and number of electron dense inclusions in the chloroplasts of plants regenerated at optimised level of copper sulphate (0.5µM) in the medium. There was decrease in chlorogenic acid (CGA) content. Chl-a-fluorescence transient pattern (OJIP) showed that the photosynthesis process was more efficient at 0.5µM CuSO4 in the medium. 

Cytotoxic effect of double emulsion (w/o/w) cuso4 loaded pla nanoparticles on mkn-45 gastric adenocarcinoma cell line / Efecto citotóxico de nanopartículas de pla de doble emulsión (w/o/w) cargadas con cuso4 sobre la línea célular de adenocarcinoma gástrico mkn-45

Poly (L-lactic acid) (PLA) nanoparticles have the approval of the main institutions for drugs administration and therapeutics. However, the use of lactic acid polymer is controversial because lactic acid has been proposed as an energy source for cancer cells. The aim of this study was to evaluate the cytotoxic, apoptotic and cell cycle properties of PLA and CuSO4-loaded PLA biodegradable nanoparticles on MKN-45 gastric adenocarcinoma cell line. PLA nanoparticles for the delivery of the anticancer active principle CuSO4 were obtained using the double emulsion method. PLA and CuSO4 loaded PLA nanoparticles were morphologically characterized and their size determined using transmission electron microscopy (TEM). The cytotoxicity of this drug delivery system was evaluated using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay; apoptosis was evaluated using YO-PRO-1/Propidium Iodide and cell cycle analysis throughout flow cytometry. CuSO4-loaded PLA nanoparticles were effective inhibitors of MKN45 cancer cell growth. They increased cytotoxicity and apoptosis, and induced G1/Go cell cycle arrest;whereas the anticancer activity was increased using a 96 h treatment of a minimal (1mM) concentration of CuSO4 loaded in 40 µM PLA nanoparticles. The treatment with 40 µM lactic acid and PLA (40 µM) did not increase the rate of cell survival assays related to the control, which indicate that PLA use as a polymer carrier not induce proliferation of MKN-45 cancer cells. Our research presents novel data about the effect of PLA nanoparticles and CuSO4 on gastric cancer cell line MKN45.

Las nanopartículas de ácido poli L-láctico (PLA) tienen la aprobación de las principales instituciones de administración de medicamentos y terapéutica. Sin embargo, el uso de polímero de ácido láctico es controvertido ya que el ácido láctico se ha propuesto como una fuente de energía para las células cancerosas. El objetivo de este estudio fue evaluar las propiedades citotóxicas, la apoptosis y sobre el ciclo celular de las nanoparticulas de PLA biodegradable y de estas PLA nanopartículas cargadas con CuSO4 en la línea celular de adenocarcinoma gástrico MKN-45. Las nanopartículas de PLA para la administración del principio activo CuSO4 contra el cáncer se obtuvieron utilizando el método de doble emulsión. Las nanopartículas de PLA y PLA cargadas con CuSO4 se caracterizan morfológicamente y su tamaño fue determinaron usando microscopía electrónica de transmisión (TEM). Se evaluó la citotoxicidad de este sistema de administración de fármacos utilizando la 3 - (4,5-dimetiltiazol-2-il) -2,5-ensayo difeniltetrazolio (MTT); la apoptosis se evaluó usando yoduro de propidio/YO-PRO-1 y el análisis de ciclo celular por citometría de flujo. Las nanopartículas cargadas con CuSO4-PLA fueron eficaces inhibidores del crecimiento de las células MKN-45 cancerosas. Aumentaron citotoxicidad y la apoptosis, e inducen la detención del ciclo celular en G1/Go, mientras que la actividad contra el cáncer se incrementó con el uso de un tratamiento de 96 horas con una concentración mínima (1 mM) de CuSO4 cargado en nanopartículas con 40 µM de PLA. El tratamiento con 40 µM de ácido láctico y 40 µM PLA no aumentó la tasa de supervivencia de células en relación con el control, lo que indica que el uso de PLA como un polímero portador que no induce la proliferación de células de cáncer MKN-45. Nuestro estudio presenta nuevos datos sobre el efecto de las nanopartículas de PLA con CuSO4 en la línea celular de cáncer gástrico MKN-45.

Genome-Wide Identification and Characterization of Novel Laccase Genes in the White-Rot Fungus Flammulina velutipes

The aim of this study was to identify and characterize new Flammulina velutipes laccases from its whole-genome sequence. Of the 15 putative laccase genes detected in the F. velutipes genome, four new laccase genes (fvLac-1, fvLac-2, fvLac3, and fvLac-4) were found to contain four complete copper-binding regions (ten histidine residues and one cysteine residue) and four cysteine residues involved in forming disulfide bridges, fvLac-1, fvLac-2, fvLac3, and fvLac-4, encoding proteins consisting of 516, 518, 515, and 533 amino acid residues, respectively. Potential N-glycosylation sites (Asn-Xaa-Ser/Thr) were identified in the cDNA sequence of fvLac-1 (Asn-454), fvLac-2 (Asn-437 and Asn-455), fvLac-3 (Asn-111 and Asn-237), and fvLac4 (Asn-402 and Asn-457). In addition, the first 19~20 amino acid residues of these proteins were predicted to comprise signal peptides. Laccase activity assays and reverse transcription polymerase chain reaction analyses clearly reveal that CuSO4 affects the induction and the transcription level of these laccase genes.

Antimicrobial evaluation of copper sulfate (II) on strains of Enterococcus faecalis: in vitro study / Evaluación antimicrobiana del sulfato de cobre (II) sobre cepas de Enterococcus faecalis: estudio in vitro

Introduction: Controlling Enterococcus faecalis is of vital importance in endodontics, as this pathogen is associated with endodontic failure. Experimental evidence has shown that copper has antibacterial activity against other pathogens with similar characteristics. The objective of this study was to determine the antimicrobial activity of copper (II) or cupric (SC-II) sulfate on strains of Enterococcus faecalis and to compare it with the most commonly used antimicrobials. Methodology: We used 33 strains of Enterococcus faecalis isolated from different clinical pictures in different Chilean hospitals. The minimum inhibitory concentration (MIC) of SC-II, chlorhexidine and calcium hydroxide was determined by the broth microdilution technique, following the recommendations given by the Clinical and Laboratory Standards Institute. Results: The MIC for CHX varied in the range of 5-10 ug/ml; SC-II from 1.5 to 12 ug/ml, and HC was >32 mg/ml. The geometric mean of SCII was 6 ug/ml, lower than that of CHX, which was 7.29 ug/ml. Conclusions: SCII showed antimicrobial activity at lower concentrations than CHX. HC (which could have been affected by the buffer effect of the broth microdilution technique) showed high values, not comparable to other compounds. We suggest carrying out further studies on the properties of SC-II, such assessing its biocompatibility and reaction with other materials to be used clinically in endodontic therapy.

Introducción: El control de Enterococcus faecalis es de vital importancia en endodoncia, ya que este patógeno está asociado al fracaso endodóntico. Evidencias experimentales que han demostrado que el cobre presenta actividad antibacteriana en otros patógenos de similares características. El objetivo de este estudio es determinarla actividad antimicrobiana del sulfato de cobre (II) o cúprico (SC-II) sobre cepas de Enterococcus faecalis y compararla con los antimicrobianos más usados en la actualidad. Metodología: Estudio in vitro. Se utilizó la técnica de microdilución en caldo según lineamientos del Clinical and Laboratory Standards Institute, incluyendo 33 cepas de Enterococcus faecalis obtenidas desde hospitales chilenos, para cada una de las cuales se determinó las concentraciones mínimas inhibitorias (CMI) de: SC-II, Clorhexidina (CHX) e Hidróxido de calcio (HC). Resultados: La CMI para CHX tuvo un rango de 5-10 ug/ml, el SC-II de 1,5-12 mM y el HC fue >32 mg/ml. Estas diferencias fueron estadísticamente significativas entre los 3 antimicrobianos utilizados (p<0,001).Conclusiones: El SC-II mostró actividad antimicrobiana a bajas concentraciones, superiores a CHX, pero menores a HC (que pudo ser afectado por el efecto tampón de la técnica de microdilución en caldo). Se sugiere seguir con los estudios de las propiedades del SC-II, como evaluación de biocompatibilidad y reacción con otros materiales para ser utilizados clínicamente en la terapia endodóntica.

Sanitizer residues in the milk of cows which had passed through footbaths / Resíduos de sanitizantes em vacas de leite após passagem por pedilúvio

Copper sulfate and sodium hypochlorite are used in footbath solutions for the prevention and treatment of bovine digital diseases; however, data on the residues of such elements in milk are sparse in Brazil. This study evaluated the cost of applying the footbath treatment and the total amount of copper and chlorite residues in the milk of healthy cows after they had passed through these footbath solutions. Two groups of 7 cows each (GI and GII) were studied. In the case of GI, 1% sodium hypochlorite was used and for GII 5% copper sulfate was employed in the footbath. The milk samples were collected before the 7-day footbath treatment period (M0) and 24 h (M1), 48 h (M2), 72 h (M3) and 15 days (M15) after the last footbath. Statistical analysis to compare the different samples within each group was carried out by applying Friedman's test, followed by Dunn's test (p<0.05). It was concluded that the amount of total chlorites and copper in the milk of healthy cattle after routine daily footbaths for a period of 7 days presented some variations. However, the concentrations observed were considered insufficient to represent a risk to human health. The cost of the footbath solutions was found to be reasonable.

O sulfato de cobre e o hipoclorito de sódio são empregados na prevenção e tratamento das enfermidades digitais dos bovinos, mas os valores residuais desses elementos foram pouco estudados. Neste estudo, avaliou-se a presença de resíduos de cobre e cloretos totais no leite de vacas saudáveis após passagens dos animais em pedilúvio contendo soluções formuladas com estas substâncias e estimou-se os custos das soluções. Utilizou-se 14 vacas saudáveis distribuídas em dois grupos (GI e GII) de sete animais cada. Em GI, empregou-se solução de hipoclorito de sódio a 1% e, em GII, sulfato de cobre a 5%. As amostras de leite foram colhidas antes da passagem pelo pedilúvio (M0), após 24 (M1), 48 (M2) e 72 (M3) horas, além de 15 dias (M15) subsequentes à última passagem. Na análise estatística, a comparação entre momentos dentro de cada grupo foi realizada com teste de Friedman, seguido pelo teste de Dunn's (p<0,05). Concluiu-se que os valores de cloretos totais e de cobre no leite de bovinos saudáveis, após passagens diárias dos animais em pedilúvio por um período de sete dias, apresentaram algumas variações consideradas insuficientes para provocarem danos à saúde humana e as soluções medicamentosas não apresentaram custos exorbitantes.

Efeitos do tratamento com formalina e sulfato de cobre sobre os parâmetros hematológicos e parasitos monogenéticos em juvenis de Hemigrammus sp. (Osteichthyes: Characidae) / Evaluation of formalin and copper sulfate on monogenetic parasite (Urocleidoides sp.) of Hemigrammus sp. (Osteichthyes: Characidae) juveniles

The objective of this study was to evaluate the effectiveness of formalin and copper sulphate to controlm onogeneans injuvenile Hemigrammus spand its effect on leukocyte count. We used acompletely randomized design with five treatments and three replications. All treatments lasted for three days and consisted of: T1, control; T2 and T3 immersion bath for one hour in formalin, an of two concentrations of 0.25ml formalin and the other 0.1mlL-1; the T4 bath for 24 hours with 0,025 ml of formalin-L -1; and T5, bath 24 hours with copper sulphate (0.3 mgL-1). After the experimental period was observed that T2, 100% mortality. The T3 and T5 provided 100% efficiency, however, fish presented mortalities and intoxication of the 66 and 80% respectively. T4 showed an efficacy of 77.7% and decrease in parasitological indices: total number of parasites of 11, 3 ± 9, mean infection intensity of 3.2 ± 2 and a prevalence of 47.1%. As the leukocytes was observed that at T3, T4 and T5 occurred differences in the proportionof lymphocytes, monocytes and neutrophilsas compared to control. The CuSO4 and formalin reduce the parasite load, but presented to be toxicat higher concentrations.

O objetivo deste trabalho foi avaliar a eficácia da formalina e sulfato de cobre no controle de monogenéticos em juvenis de Hemigrammus sp e seu efeito na contagem diferencial de leucócitos. Utilizou-se um delineamento inteiramente casualizado com 5 tratamentos e 3 repetições. Todos os tratamentos tiveram duração de três dias e corresponderam a: T1, controle; T2 e T3 banho por imersão de uma hora em formalina, em duas concentrações uma de 0,25 ml e outra 0,1 ml formalina L-1; o T4, banho de 24 horas com 0,025 ml de formalina L-1; e o T5, banho de 24 horas com sulfato de cobre (0,3 mg L-1). Após o período experimental observou-se que em T2 ocorreu 100% de mortalidade. O T3 e T5 proporcionaram eficácia de 100%, porém os peixes apresentaram intoxicação e mortalidades de 66 e 80%, respectivamente. T4 apresentou eficácia de 77,7% e diminuição dos índices parasitológicos: número total de parasitos de 11,3± 9, intensidade média de infecção de 3,2± 2 e prevalência de 47,1%. Quanto aos leucócitos observou-se que no T3, T4 e T5 ocorreram diferenças na proporção de linfócitos, monócitos e neutrófilos em relação ao controle. O CuSO4 e a formalina reduzem a carga parasitária, mas apresentaram-se tóxicos nas maiores concentrações.

La intoxicación con cobre disminuye la sobrevida e induce alteraciones neurológicas en Drosophila melanogaster / Copper intoxication decreases lifespan and induces neurologic alterations in Drosophila melanogaster

La enfermedad de Wilson, es un trastorno hereditario autosómico recesivo causado por mutaciones del gen de la trifosfatasa de adenosina (ATP7B). Dicha mutación ocasiona intoxicación con cobre, generando manifestaciones clínicas por los efectos tóxicos del metal, principalmente a nivel del hígado y el encéfalo. Recientemente se han desarrollado modelos genéticos de la enfermedad para su estudio clínico. Sin embargo, la utilidad de los mismos es limitada por el hecho de que en tales modelos no se observan manifestaciones neurológicas. El presente estudio tuvo como objetivo desarrollar un modelo de la enfermedad de Wilson en Drosophila melanogaster. Inicialmente se evaluó el efecto de la suplementación con concentraciones de 31 µM y 47 µM de cobre en la sobrevida. Posteriormente se realizaron estudios de conducta para determinar si existían alteraciones en el desempeño motor asociadas al tratamiento con la dosis de 47 µM de cobre. Los resultados obtenidos sugieren que el tratamiento con cobre disminuye la viabilidad de la Drosophila. La disminución de la sobrevida estuvo asociada a un aumento y una disminución de los registros de actividad motora en las etapas tempranas y tardías de la intoxicación respectivamente. Por último, se evaluó el papel del sistema de neurotransmisión dopaminérgico sobre las alteraciones conductuales inducidas por el cobre. El tratamiento con el precursor de la dopamina, L-dopa, indujo un aumento de la actividad motora similar al inducido por el cobre. Por el contrario, el tratamiento con Flufenazina, un antagonista de los receptores dopaminérgicos D2, fue capaz de impedir las alteraciones conductuales en todas las edades evaluadas. Estos resultados sugieren que la Drosophila melanogaster podría ser empleada como modelo para el estudio de posibles intervenciones con potencial terapéutico en la enfermedad de Wilson.

Wilson disease is a hereditary disorder caused by mutations of the ATP7B gene, which leads to intoxication with copper as a result of an unbalance of copper homeostasis. The clinical manifestations resulting from this intoxication are related to the affectation of liver and the encephalon in most cases. Several animal models are currently available for the study of the malady. However, in such models no neurological symptoms are observed, which limits their use for the study of pathogenic effects of this disease on the central nervous system. The aim of the present study was to evaluate if copper feeding could induce a disease state in Drosophila melanogaster to model Wilson disease. The effect of the feeding of copper at the doses of 31 µM and 47 µM on the survival was initially evaluated. Next, behavioral experiments were conducted to determine whether the motor performance was altered by the 47 µM concentration. The results suggest that copper treatment decreases the viability of the flies. In addition, the decrease of viability was associated to an increase and decrease of spontaneous motor activity at early and late stages of the intoxication, respectively. Finally, the role of the dopaminergic neurotransmission system on the observed motor alterations was evaluated. The dopamine precursor L-dopa increased motor activity. In contrast, D2 receptor antagonist, Fluphenazine, was able to block both the increase and decrease of motor activity scores induced by copper. These results suggest that Drosophila melanogaster could be used as a model organism for the study of possible interventions with potential neuroprotective effects in Wilson disease.

Zinc therapy improves adverse effects of long term administration of copper on epididymal sperm quality of rats

Industrial copper ingest is a common form of poisoning in animals. Zinc has an important role in the physiology of spermatozoa, in sperm production and viability. This study was set to investigate whether the adverse effects of long term copper consumption on quality of rat spermatozoa could be prevented by zinc therapy. Forty eight mature [6-8 weeks old] male rats were randomly allocated to either control [Cont, n=12] or three treatment groups each containing twelve animals. Animals in the first treatment group was gavaged with copper sulfate, the second treatment group was injected with zinc sulfate, and the third treatment group was given combined treatment of copper and zinc. Control animals received normal saline using the same volume and similar methods. Six rats from each group were sacrificed on day 28 and 56 after treatments for sperm quality evaluations. In spite of testicular weight reduction 56 days after copper consumption in comparison to the control group [p=0.002], there was not a significant difference between the control and combined treatment of copper and zinc group [31.40 +/- 0.55 vs. 28.63 +/- 0.55, p=0.151]. Administration of copper caused a significant decrease in the sperm count, viability and motility after 56 days compared to the control group. However, a complete recovery in sperm count was seen in combined treatment of copper and zinc group after 56 days compared to the control group [p=0.999] and a partial improvement was seen about the percentage of viability and motility [p<0.001]. Adverse effects of long term consumption of copper on sperm quality could be prevented by zinc therapy in rats

Antioxidant properties and inhibitory effects of Satureja khozestanica essential oil on LDL oxidation induced-CuSO(4) in vitro

<p><b>OBJECTIVE</b>To assess various antioxidative activities of Satureja khozestanica essential oil (SKE) and its effect on oxidation of low density lipoprotein (LDL) induced by CuSO4 in vitro by monitoring the formation of conjugated dienes and malondialdehyde (MDA).</p><p><b>METHODS</b>The formation of conjugated dienes, lag time and MDA were measured. Inhibition of this Cu-induced oxidation was studied in the presence of several concentrations of SKE. Also total antioxidant activity and free radical scavenging of SKE were determinated.</p><p><b>RESULTS</b>It was demonstrated that SKE was able to inhibit LDL oxidation and decrease the resistance of LDL against oxidation. The inhibitory effects of SKE on LDL oxidation were dose-dependent at concentrations ranging from 50 to 200 µg/mL. Total antioxidant capacity of SKE was (3.20±0.40) nmol of ascorbic acid equivalents/g SKE. The SKE showed remarkable scavenging activity on 2, 2-diphenyl-picrylhydrazyl, IC50 (5.30±0.11) ng/mL.</p><p><b>CONCLUSIONS</b>This study shows that SKE is a source of potent antioxidants and prevents the oxidation of LDL in vitro and it may be suitable for use in food and pharmaceutical applications.</p>

Actividad de cobre sobre bacilos gramnegativos multi-resistentes aislados en hospitales chilenos / Copper activity against multiresistant Gram negative bacilli isolated from Chilean hospitals

Introduction: Multiresistant nosocomial pathogens, especially Gram-negative bacilli (GNB), are a serious problem for public health systems worldwide. Due to their antimicrobial properties, copper alloys have been suggested as an alternative for the control of bacterial burden in surfaces in hospital environment. However, antibiotic multiresistance and copper resistance could be associated in GNB, and there is evidence that both kind of resistance genes (antibiotic and copper) can be located on the same genetic structures. For this reason antibiotic-multiresistant strains could survive in the presence of copper, selecting for bacterial phenotypes resistant to both antibacterial agents. Aim: To evaluate antibacterial activity of copper against nosocomial extended-spectrum β-lactamases (ESBL) (+) and ESBL (-) GNB, and carbapenems resistant or susceptible strains. Material and Method: This study included 390 strains of GNB isolated from Chilean hospitals: Acinetobacter baumannii and Pseudomonas aeruginosa resistant (CAR R) and susceptible (CAR S) to carbapenem antibiotics, and Klebsiella pneumoniae and Escherichia coli producers and non-producers of ESBL. Susceptibility levels to cupric sulphate were determined by agar dilution method and statistical analysis were used to determine the significance of the differences in the copper tolerance levels between the strains groups. Results: Statistically superior copper tolerance levels were found in the CAR R and ESBL producing strains of A. baumannii and K. pneumoniae, in relation with the CAR S and ESBL not-producing strains. Conclusion: A relation between a diminished susceptibility to ionic copper and to recent generation antimicrobial agents was observed in K. pneumoniae y A. baumannii strains.

Introducción: Los patógenos intrahospitalarios multi-resistentes constituyen un grave problema mundial de salud pública, especialmente los bacilos gramnegativos (BGN). El uso de cobre como antimicrobiano de superficie en hospitales se postula como una alternativa para el control de microorganismos en estos ambientes. Sin embargo, la multi-resistencia a antimicrobianos en BGN hospitalarios puede asociarse con la tolerancia a cobre, ya que existe evidencia que genes que codifican tolerancia a este metal pueden encontrarse en elementos genéticos que confieren resistencia a antimicrobianos. Por esta razón, cepas multi-resistentes a antimicrobianos podrían sobrevivir en presencia de cobre, seleccionando bacterias resistentes a ambos agentes antibacterianos. Objetivo: Investigar la actividad de cobre sobre BGN hospitalarios productores y no productores de β-lactamasas de espectro extendido (BLEE), y resistentes o susceptibles a antimicrobianos carbapenémicos. Material y Métodos: Se estudió 390 cepas de BGN aisladas en hospitales chilenos: Acinetobacter baumannii y Pseudomonas aeruginosa resistentes (CAR R) y susceptibles (CAR S) a carbapenémicos y Klebsiella pneumoniae y Escherichia coli productoras y no productoras de BLEE. Se investigó los niveles de susceptibilidad a sulfato cúprico, mediante dilución seriada en agar y se evaluó la significancia estadística de la diferencia de estos niveles entre los distintos grupos de cepas. Resultados: Se encontraron niveles de tolerancia a cobre superiores en cepas de A. baumannii y K. pneumoniae, CAR R y productoras de BLEE respectivamente, con respecto a sus pares CAR S y no productoras de BLEE. Conclusión: Observamos una relación entre la disminución de la susceptibilidad a cobre iónico y a antimicrobianos de última generación en K. pneumoniae y A. baumannii.

Effects of olive leaves extract on LDL oxidation induced-CUSO[4] in vitro

Oxidation of low-density lipoprotein [LDL] has been strongly implicated in the pathogenesis of atherosclerosis. The use of some natural antioxidant and herbal medicine may lead to the inhibition of production of oxidized LDL and may decrease both the development and the progression of atherosclerosis. The aim of this study was to investigate the effects of Olive leaves ethanol extract [OLE] on LDL oxidation induced-CuSO[4] quantitatively in vitro. Low-density lipoprotein was incubated with CuSO[4] and the formation of conjugated dienes and thiobarbituric acid reactive substances [TBARS]. Inhibition of this Cu-induced oxidation was studied in the presence of vitamin E and various concentration of OLE. It was demonstrated that OLE reduced the formation of conjugated dienes and TBARS of LDL against oxidation in vitro [P<0.05]. The inhibitory effects of the OLE on LDL oxidation were dose-dependent at concentrations ranging from [2microg/ml] to [200microg/ml]. Moreover, we compared effects of OLE on LDL oxidation with vitamin E as positive control. This study showed that OLE is a source of potent antioxidants and prevented the oxidation of LDL in vitro and it may be suitable for use in food and pharmaceutical applications