Role of Silver Nitrate Spray for Skin Wound Care in Patients with Toxic Epidermal Necrolysis: Our Experience in 4 Patients.

Stevens-Johnson syndrome (SJS) and toxic epidermal necrolysis (TEN) are examples of severe cutaneous adverse reactions to drugs (SCARs) with several international recommendations for global medical management, ranging from pharmacological systemic therapy to skin wound care. There is no defined best management of the skin wounds in SJS/TEN. The care of wounds is essential to initiate re-epithelialization. Our objective is to improve the cicatrization process, avoiding scarring due to deepening of the wounds, as well as prevent infections, achieve pain control, and avoid loss of serum proteins, fluids, and electrolytes. In this retrospective case series, we highlight the value of systemic therapy and the use of silver nitrate for wound management in four patients with TEN.

Silver Nitrate Staining of Nucleolar Organizer Regions (Ag-NORs) in Plant Chromosomes.

Silver nitrate staining to evidence the location of nucleolar organizer regions (Ag-NORs) in chromosomes is widely used as a classical method in plant cytogenetics. Here, we present the most used procedures and highlight some aspects in terms of their replicability by plant cytogeneticists. Some technical features described are materials and methods used, procedures, protocol modifications, and precautions in order to obtain positive signals. The methods to obtain Ag-NOR signals have different degrees of replicability, but do not require any sophisticated technology or equipment for their application.

Metabolic response induced by abiotic elicitors in Anacardium othonianum seedlings.

Applying elicitors in plants growing under natural conditions change the metabolic content of volatile compounds extracted from their tissues. Here, we have investigated how abiotic elicitors influence the volatile composition of the ethyl acetate extract of Anacardium othonianum seedlings propagated in vitro and ex-vitro cultures. Abiotic factors such as salicylic acid, silver nitrate, light quality, and cultivation mode affected the composition of volatile compounds in ethyl acetate extract of A. othonianum leaves seedlings. GC/MS analysis demonstrated that α-tocopherol was the main metabolite in cultures added with chemical elicitors, reaching 22.48 ± 4.06% in the case of 30 µM salicylic acid. Higher concentrations of salicylic acid decreased the amount of α-tocopherol. The addition of AgNO3 had a linear effect on α-tocopherol. Thus, the production of this metabolite indicates that salicylic acid and AgNO3 elicitors positively affect α-tocopherol production at specific concentrations helping to understand how plant culture can be better manipulated.

Multiplication, morphogenesis and foliar abscision of Schinopsis brasiliensis Engl. in vitro / Multiplicación, morfogénesis y abscisión foliar de Schinopsis brasiliensis Engl. in vitro

The objective was to evaluate plant growth regulators and ethylene inhibitors on the development and leaf abscission of Schinopsis brasiliensis Engl. Zeatin (ZEA) was evaluated in concentrations combined with concentrations of indolacetic acid (IAA), naphthalene acetic acid (NAA) and indolbutyric acid (IBA). ZEA and 6-benzylamino purine (BAP) were evaluated in concentrations plus a control. Ethylene inhibitors, silver nitrate and cobalt chloride were evaluated in four concentrations. The addition of 0.2 µL-1 of NAA to 0.4 µL-1 of ZEA promotes a greater number of baraúna sprouts. At concentrations of 5 and 10 µM, cobalt chloride is more efficient than silver nitrate for reducing leaf abscission in baraúna. IAA is the most suitable auxin to be associated with ZEA for higher shoot length and number of buds. Silver nitrate from a concentration of 20 µM completely avoids leaf abscission whilecobalt chloride has a maximum reduction in abscission at a concentration of 40 µM.

El objetivo fue evaluar reguladores de crecimiento e inhibidores de etileno sobre el desarrollo y abscisión foliar en Schinopsis brasiliensis Engl. La zeatina (ZEA) se evaluó en concentraciones combinadas con concentraciones de ácido indolacético (IAA), ácido naftaleno acético (NAA) y ácido indolbutírico (IBA). Se evaluaron ZEA y 6-bencilamino purina (BAP) en concentraciones más un control. Se evaluaron inhibidores de etileno, nitrato de plata y cloruro de cobalto, en cuatro concentraciones. La adición de 0.2 µL-1 de NAA a 0.4 µL-1 de ZEA promueve un mayor número de brotes de baraúna. A concentraciones de 5 y 10 µM, el cloruro de cobalto es más eficaz que el nitrato de plata para reducir la abscisión de las hojas en baraúna. IAA es la auxina más adecuada para asociar con ZEA para una mayor longitud de brotes y número de brotes. El nitrato de plata a partir de una concentración de 20 µM evita completamente la abscisión de las hojas, mientras que el cloruro de cobalto tiene una reducción máxima en la abscisión a una concentración de 40 µM.

Obtaining Ultrafine Dispersions of Silver Particles in Poly(vinyl Alcohol) Matrix Using Mechanical Alloying.

Mechanical alloying was performed to obtain a composite material with a homogeneous dispersion of silver particles in a poly(vinyl alcohol) (PVA) matrix. Silver is a bactericidal material, and PVA is a widely used biocompatible polymer. Therefore, this mix can lead to a potentially functional biomaterial. This study focuses on the combination of both materials, processed by mechanical alloying, which has a promising application potential. The silver (Ag) used was ultrafine, measuring between 200 and 400 nanometers, produced from silver nitrate (AgNO3) redox. The Attritor high-energy, water-cooled ball mill was used to mill PVA for 4 h, at 600 rpm speed rotation and 38:1 power milling. Mechanical alloying was demonstrated to cause particle refinement in PVA with a timespan of 1 h. A slight additional particle decrease occurred for long-time milling. A milling time of 4 h was used to disperse the silver particles in the polymer matrix homogeneously. Hot pressing films were produced from the obtained dispersion powders. The microstructural features were studied using several material characterization techniques. Antimicrobial Susceptibility Tests (AST), conducted in an in-vitro assay, showed a hydrophilic character of the films and a protection against bacterial growth, making the process a promising path for the production of surface protective polymeric films.

In vitro antileishmanial activity of sustainable anacardic acid and cardol based silver nanoparticles on L. braziliensis.

The search for effective and less toxic drugs for the treatment of Cutaneous Leishmaniasis (CL) is desirable due to the emergence of resistant parasites. The present study shows the preparation, characterization and in vitro antileishmanial activity of green-based silver nanoparticles (AgNPs) with Cashew Nutshell Liquid (CNSL, main constituents: anacardic acid (AA) and cardol (CD). The synthesis of silver nanoparticles was achieved by reduction with sodium borohydride in the presence of anacardic acid or cardol under microwave irradiation (400 W, 60 °C, 5 min) resulting in AgAA and AgCD. In vitro assay showed opposite effects for AgAA and AgCD. While AgAA is highly toxic to macrophages (CC50 = 6.910 µg mL-1) and almost non-toxic for L.braziliensis (IC50 = 86.61 µg mL-1), AgCD results very selective toward killing the parasite (CC50 = 195.0 µg mL-1, IC50 = 11.54 µg mL-1). AA's higher polarity and conical shape easily promote cell lysis by increasing cell permeability, while CD has a protective effect: for that reason, AA and AgAA were not further used for tests. CD (EC50 = 2.906 µg mL-1) had higher ability to kill intracellular amastigotes than AgCD (EC50 = 16.00 µg mL-1), however, less intact cells were seen on isolated CD tests. In addition, considering that NO is one of the critical molecular species for the intracellular control of Leishmania, we used Griess colorimetric test to analyze the effect of treatment with AgCD and CD. Overall, the in vitro antileishmanial tests indicate that AgCD should be further explored as a promising non-toxic treatment for CL.

Microleakage in class II restorations of two bulk fill resin composites and a conventional nanohybrid resin composite: an in vitro study at 10,000 thermocycles.

BACKGROUND: The contraction presented by resin composites causes an increase in stress at the tooth-resin interface, causing micro-gaps that allow microleakage. This study aims to evaluate the degree of in vitro marginal microleakage in class II restorations with two bulk fill resin composites compared to a conventional nanohybrid resin composite. METHODS: The present study was an in vitro experimental design. A total of 30 standardized class II cavities were prepared in 15 human molars (mesially and distally). These cavities were later distributed in 3 groups according to the type of resin. Groups A and B were restored with bulk fill resin composites (Filtek-3 M/ESPE and Tetric N-Ceram-Ivoclar/Vivadent respectively) in a single increment of 4 mm. Group C was restored with the Filtek Z350 XT - 3 M/ESPE resin composite and two increments of 2 mm. Later, the restorations were subjected to 10,000 thermocycles between 5 °C to 55 °C and immersed in a silver nitrate solution (1 M for 24 h). The crowns were then sectioned mesiodistally and observed under the stereomicroscope to determine the degree of marginal microleakage at the occlusal and cervical areas. The results were analyzed with the Kruskal-Wallis and the Mann-Whitney U statistical tests. RESULTS: There were no statistically significant differences regarding the degree of microleakage between the three types of resin composites in the occlusal and cervical areas (p > 0.05). Similarly, there were no significant differences after comparing each resin type in its occlusal and cervical area (p > 0.05). CONCLUSION: Filtek Bulk Fill and Tetric N-Ceram Bulk Fill resin composites showed no statistically significant differences with the conventional nanohybrid resin composite Filtek Z350XT at both occlusal and cervical areas.

Exogenous pigmentation by silver nitrate: Dermatological and toxicological aspects, case report / [Pigmentación exógena por nitrato de plata: aspectos dermatológicos y toxicológicos, a propósito de un caso].

Exogenous pigmentation by silver nitrate is a rare disease whose clinical manifestations appear even years after the contact, making its diagnosis difficult on occasions. It is characterized by the presence of blue-gray macules or plaques on the skin or mucosa in the contact area, sometimes very similar to melanocytic lesions and melanoma, which constitute the main differential diagnosis. We report the case of a male patient from Medellín, Colombia, with a family history of melanoma and the presence of these lesions throughout his body.

La pigmentación exógena por nitrato de plata es una enfermedad poco frecuente, cuyas manifestaciones clínicas pueden aparecer años después del contacto, lo que en ocasiones dificulta su diagnóstico. Se caracteriza por la presencia de máculas o placas azul-grisáceas en la piel o las mucosas de la zona de contacto que, en ocasiones, son muy similares a las lesiones melanocíticas y al melanoma, sus principales diagnósticos diferenciales. Se reporta el caso de un paciente de Medellín, Colombia, con antecedentes familiares de melanoma y presencia de estas lesiones en todo el cuerpo.

Pigmentación exógena por nitrato de plata: aspectos dermatológicos y toxicológicos, a propósito de un caso / Exogenous pigmentation by silver nitrate: Dermatological and toxicological aspects, case report

Resumen | La pigmentación exógena por nitrato de plata es una enfermedad poco frecuente, cuyas manifestaciones clínicas pueden aparecer años después del contacto, lo que en ocasiones dificulta su diagnóstico. Se caracteriza por la presencia de máculas o placas azul-grisáceas en la piel o las mucosas de la zona de contacto que, en ocasiones, son muy similares a las lesiones melanocíticas y al melanoma, sus principales diagnósticos diferenciales. Se reporta el caso de un paciente de Medellín, Colombia, con antecedentes familiares de melanoma y presencia de estas lesiones en todo el cuerpo.

Abstract | Exogenous pigmentation by silver nitrate is a rare disease whose clinical manifestations appear even years after the contact, making its diagnosis difficult on occasions. It is characterized by the presence of blue-gray macules or plaques on the skin or mucosa in the contact area, sometimes very similar to melanocytic lesions and melanoma, which constitute the main differential diagnosis. We report the case of a male patient from Medellín, Colombia, with a family history of melanoma and the presence of these lesions throughout his body.

Nanopartículas de prata em bases de próteses de PMMA para controle de atividade microbiana / Silver nanoparticles in PMMA prosthesis bases to control microbial activity

Objetivo:Avaliar in vitro a atividade de Staphylococcus aureus e Candida albicans em bases de próteses convencionais à base de polimetilmetacrilato de metila com nanopartículas de prata incorporadas a sua composição. Métodos: Foi realizado um estudo experimental laboratorial com resinas acrílicas autopolimerizáveis comercialmente disponíveis, Vipi Flash/VIPI e JET/Clássico. Foram confeccionados 80 corpos de prova, divididos em 16 grupos (n = 5), referentes ao tipo de resina, tratamento (incorporação e imersão na solução de nanopartículas de prata) e microrganismo inoculado. As nanopartículas foram sintetizadas com ácido polimetacrílico, nitrato de prata e irradiadas com luz ultravioleta de baixa potência (~8W) por 6 horas, e as suas concentrações idealizadas pelo método de microdiluição em placas para determinação da concentração mínima inibitória frente aos microrganismos selecionados. Verificou-se ação bactericida e fungicida com concentração inicial de 25% e após fator de diluição 12,5%. Resultados: Houve dificuldade de incorporação das nanopartículas na resina acrílica, que pode decorrer da alteração da proporção 3:1 recomendada pelo fabricante ou pela redução ou inativação da ação da nanopartícula de prata pela interação com o polimetilmetacrilato. VIPI com inclusão de nanopartícula obteve menor aderência de biofilme de Candida albicans. Conclusão:A nanopartícula de prata mostrou-se eficaz na sua ação de controle de Candida albicans e Staphylococcus aureus no método de imersão, entretanto, a sua ação antimicrobiana foi comprometida após inclusão nas resinas acrílicas.

Aim:To perform an in vitro evaluation of the activity of Staphylococcus aureus and Candida albicansin conventional prosthesis bases, based on methyl polymethylmethacrylate with silver nanoparticles incorporated into the composition. Methods: An experimental laboratory study was carried out using commercially available self-curing acrylic resins, Vipi Flash/VIPI and JET/Clássico. Eighty specimens were manufactured and divided into 16 groups (n = 5), referent to the resin brand, treatment (incorporation and immersion in the silver nanoparticle solution), and inoculated microorganism. The nanoparticles were synthesized with polymethacrylic acid and silver nitrate, and were irradiated with a low power (~ 8W) ultraviolet light for 6 hours. Their concentrations were idealized by the method of microplate dilution to determine the minimum inhibitory concentration when compared to the selected microorganisms. Bactericidal and fungicidal activities were identified with an initial concentration of 25% and a subsequent dilution factor of 12.5%. Results:It was difficult to incorporate the AgNPs into the acrylic resin, which may well have resulted from the change from the 3:1 proportion recommended by the manufacturer or by reducing or inactivating the action of the silver nanoparticle by interaction with polymethylmethacrylate. VIPI with the inclusion of nanoparticles obtained a lesser Candida albicans biofilm adherence. Conclusion: Silver nanoparticles were effective in controlling Candida albicans and Staphylococcus aureus in the immersion method; however, the antimicrobial activity was compromised after inclusion in acrylic resins.

Histological comparison of two special methods of staining melanin in human skin / Comparación histológica de dos métodos especiales de tinción de melanina en piel humana

SUMMARY: Human skin melanin was stained using the Fontana's silver nitrate method and Schmorl method. The results showed that, in the Fontana's silver nitrate method, melanin and silver-bound cells were black and other tissues were red. When stained using the Schmorl method, effects on melanin differed based on whether the nuclei were stained. When the nucleus was stained, melanin appeared blue-black or blue-green, and other tissue structures were purple. When the nucleus was not stained, melanin was orange and other structures were pink. Comparing the two staining methods, we concluded that Fontana's silver nitrate method takes a long time; in contrast, the Schmorl method showed two different types of results depending on whether the nucleus was stained, and it takes less time than Fontana staining, so we here consider the Schmorl method more suitable for special staining of melanin than Fontana's silver nitrate method.

RESUMEN: La melanina de la piel humana se tiñó utilizando el método del nitrato de plata de Fontana y el método Schmorl. Los resultados mostraron que, en el método del nitrato de plata de Fontana, la melanina y las células unidas a plata eran negras y otros tejidos eran rojos. Cuando se tiñó con el método de Schmorl, los efectos sobre la melanina difirieron en función de si se tiñeron los núcleos. Cuando se tiñó el núcleo, la melanina apareció de color azul-negro o azul-verde, y otras estructuras de tejido fueron de color púrpura. Cuando el núcleo no estaba teñido, la melanina era naranja y otras estructuras eran rosadas. Al comparar los dos métodos de tinción, llegamos a la conclusión de que el método del nitrato de plata de Fontana lleva mucho tiempo; por el contrario, el método Schmorl mostró dos tipos diferentes de resultados dependiendo de si el núcleo estaba teñido, y lleva menos tiempo que la tinción de Fontana, por lo que aquí consideramos que el método Schmorl es más adecuado para la tinción especial de melanina que el método del nitrato de plata de Fontana.

Antibacterial application of natural and carboxymethylated cashew gum-based silver nanoparticles produced by microwave-assisted synthesis.

This study presents a green synthesis route to silver nanoparticles (AgNPs) stabilized with cashew gum (CG) or carboxymethylated cashew gum (CCG) using microwave-assisted synthesis and evaluates their antibacterial activity. The antimicrobial activity was measured by determining the minimum inhibitory concentration (MIC) with Staphylococcus aureus and Escherichia coli. In both cases of the presence of CG and CCG, it was found that higher pH lead to more efficient conversion of silver nitrate to AgNPs with well dispersed, spherical and stable particles as well as low crystallinity. CCG-capped AgNPs were slightly smaller (137.0 and 96.3 nm) than those coated with non-modified gum (144.7 and 100.9 nm). The samples presented promising antibacterial activity, especially on Gram-negative bacteria, resulting in significant membrane damage on treated bacteria in comparison to the untreated control, observed by atomic force microscopy. Thus, a quick and efficient synthesis route was applied to produce CGAgNPs and CCGAgNPs with antimicrobial potential.

Local irrigation with silver nitrate, a non-surgical approach for persistent anal fistula / Irrigação local com nitrato de prata, uma abordagem não cirúrgica para fístula anal persistente

ABSTRACT Purpose: Treatment of persistent anal fistula implies a major challenge for surgeons, with risk of additional recurrence and potential continence impairment. We present a non-surgical treatment based on irrigation with silver nitrate 1% solution. Methods: This is a prospective study including patients with persistent anal fistula after surgery, who were treated with silver nitrate 1% solution irrigation from May 2015 to March 2017. Patients with evidence of abscess, presence of >1 external opening and those with bowel inflammatory disease were excluded. 3-5 cc of silver nitrate 1% solution were instilled through a catheter. The procedure was repeated on a weekly basis, conducting a maximum of 7 sessions per patient. Results: 18 patients (13 male, 72.2%) with a median age of 48 years old (IQR 41-55) were treated using silver nitrate 1% solution. A median of 5 sessions per patient was performed (IQR 3-6). The median follow-up period was 18 months (IQR 9-25). After the described period 8 patients' (44.4%) presented complete resolution of the fistula, 2 patients' (11.2%) were classified as partial healing and in 8 patients' (44.4%) the treatment was considered to fail. 6 patients' experienced self-limited pain during instillation, with persistence up to 24 h in 2 of them. Conclusions: Treatment with silver nitrate 1% solution is a minimally invasive procedure, with a favourable safety profile, that can be performed in an outpatients' basis achieving a complete healing rate of 44.4%. Therefore, this method should be considered for the treatment of recurrent or persistent anal fistula.

RESUMO Objetivo: O tratamento da fístula anal persistente é um grande desafio para os cirurgiões, com risco de recorrência adicional e potencial comprometimento da continência. Os autores apresentam um tratamento não cirúrgico baseado na irrigação com solução de nitrato de prata a 1%. Métodos: Este foi um estudo prospectivo incluindo pacientes com fístula anal persistente após a cirurgia que foram tratados com irrigação com solução de nitrato de prata a 1% entre maio de 2015 e março de 2017. Pacientes com evidência de abscesso, presença de mais de uma abertura externa e aqueles com doença inflamatória intestinal foram excluídos. Usando um cateter, instilou-se 3 a 5 cc. de solução de nitrato de prata a 1%. O procedimento foi repetido semanalmente, em um máximo de sete sessões por paciente. Resultados: Um total de 18 pacientes (13 homens, 72,2%) com idade mediana de 48 anos (IQR 41-55) foram tratados com solução de nitrato de prata a 1%. Uma mediana de cinco sessões por paciente foi realizada (IQR 3-6). A mediana do período de acompanhamento foi de 18 meses (IQR 9-25). Após o período descrito, oito pacientes (44,4%) apresentaram resolução completa da fístula, dois pacientes (11,2%) foram classificados como cicatrização parcial e em oito pacientes (44,4%) o tratamento falhou. Seis pacientes apresentaram dor autolimitada durante a instilação, que persistiu por até 24 horas em dois deles. Conclusões: O tratamento com solução de nitrato de prata a 1% é um procedimento minimamente invasivo, com perfil de segurança favorável, que pode ser realizado em regime ambulatorial, atingindo taxa de cura completa de 44,4%. Portanto, este método deve ser considerado para o tratamento da fístula anal recorrente ou persistente.

Effect of salicylic acid and silver nitrate on rutin production by Hyptis marrubioides cultured in vitro / Efeito do ácido salicílico e nitrato de prata na produção de rutina por Hyptis marrubioides cultivada in vitro

Hyptis marrubioides (Lamiaceae) is a medicinal plant that is native from Brazilian Cerrado. In vitro propagation techniques make use of elicitors to alter metabolic pathways, affecting how molecules are produced both qualitatively and quantitatively. This research aimed to evaluate how abiotic elicitors salicylic acid (SA) and silver nitrate (SN) at concentrations of 30µM or 60µM influence Hyptis marrubioides seedling growth by two different in vitro culture methods. The rutin content was quantified by HPLC-DAD. Compared to an untreated culture, the H. marrubioides methanolic extracts cultured in MS medium for 10 days followed by culture in MS medium containing SN (30µM) for 20 days had 1.28 times higher rutin content. In a second experiment, seedlings were cultured in MS medium for 20 days, and then the desired elicitor was added to the culture and allowed to remain in contact with the medium for three and six days. SA (30µM) gave the best results: rutin production was 16.56-foldhigher than the control after six days. SN (30µM) increased the rutin content by 1.17-fold. At the two concentrations evaluated during the elicitation experiments, neither SA nor SN altered the growth parameters shoot length, leaf number, and fresh and dry weight of H. marrubioides seedlings grown in vitro as compared to the control. Based on these results, the abiotic elicitors SA and SN successfully provide Hyptis marrubioides with increased rutin content in vitro.(AU)

RESUMO: Hyptis marrubioides (Lamiaceae) é uma planta medicinal nativa do Cerrado brasileiro. Técnicas de propagação in vitro fazem uso de elicitores para alterar as vias metabólicas, afetando a produção de moléculas qualitativa e quantitativamente. Este trabalho teve como objetivo avaliar como os elicitores abióticos ácido salicílico (SA) e nitrato de prata (SN) nas concentrações de 30µM ou 60µM influenciam no crescimento de plântulas de Hyptis marrubioides por dois diferentes métodos de cultivo in vitro. O teor de rutina foi quantificado por CLAE-DAD. Em comparação com uma cultura não tratada, os extratos metanólicos de H. marrubioides cultivados em meio MS por 10 dias, seguidos de cultura em meio MS contendo SN (30µM) por 20 dias, apresentaram 1,28 vezes maior teor de rutina. Em um segundo experimento, as plântulas foram cultivadas em meio MS por 20 dias, e então o elicitor desejado foi adicionado à cultura e deixado em contato com o meio por três e seis dias. SA (30µM) forneceu os melhores resultados na produção de rutina, sendo 16,56 vezes maior do que o controle após seis dias. SN (30µM) aumentou o teor de rutina em 1,17 vezes. Nas duas concentrações avaliadas durante os experimentos de elicitação, nem SA nem SN alteraram os parâmetros de crescimento, como comprimento da parte aérea, número de folhas ou peso fresco e seco das plântulas de H. marrubioides cultivadas in vitro em relação ao controle. Com base nestes resultados, os elicitores abióticos SA e SN forneceram com sucesso Hyptis marrubioides in vitro com maior conteúdo de rutina.(AU)

Influence of Silver Nitrate on Somatic Embryogenesis Induction in Arabica Coffee (Coffea arabica L)

Abstract The influence of silver nitrate (AgNO3), benzyladenine (BAP), and indole-3-acetic acid (IAA) on low frequency somatic embryogenesis (LFSE) induction in Caturra and Catuaí arabica coffee was evaluated. For the Caturra cultivar, the production of somatic embryos was significantly increased by adding AgNO3 to the semisolid culture medium. The highest average number of somatic embryos for this cultivar was obtained using 6.6 μM BAP, 2.85 μM IAA, and 40 μM AgNO3. In contrast, for the Catuaí cultivar, the highest average number of somatic embryos was obtained using semisolid medium supplemented with 8.8 μM BAP, and 2.85 μM IAA. Using these protocols, somatic embryos were directly induced using leaf sections of in vitro plants of both coffee cultivars within 8 weeks. The somatic embryos developed into rooted plants with a 100% survival rate upon transfer to the greenhouse.

Effect of salicylic acid and silver nitrate on rutin production by Hyptis marrubioides cultured in vitro / Efeito do ácido salicílico e nitrato de prata na produção de rutina por Hyptis marrubioides cultivada in vitro

ABSTRACT: Hyptis marrubioides (Lamiaceae) is a medicinal plant that is native from Brazilian Cerrado. In vitro propagation techniques make use of elicitors to alter metabolic pathways, affecting how molecules are produced both qualitatively and quantitatively. This research aimed to evaluate how abiotic elicitors salicylic acid (SA) and silver nitrate (SN) at concentrations of 30µM or 60µM influence Hyptis marrubioides seedling growth by two different in vitro culture methods. The rutin content was quantified by HPLC-DAD. Compared to an untreated culture, the H. marrubioides methanolic extracts cultured in MS medium for 10 days followed by culture in MS medium containing SN (30µM) for 20 days had 1.28 times higher rutin content. In a second experiment, seedlings were cultured in MS medium for 20 days, and then the desired elicitor was added to the culture and allowed to remain in contact with the medium for three and six days. SA (30µM) gave the best results: rutin production was 16.56-foldhigher than the control after six days. SN (30µM) increased the rutin content by 1.17-fold. At the two concentrations evaluated during the elicitation experiments, neither SA nor SN altered the growth parameters shoot length, leaf number, and fresh and dry weight of H. marrubioides seedlings grown in vitro as compared to the control. Based on these results, the abiotic elicitors SA and SN successfully provide Hyptis marrubioides with increased rutin content in vitro.

RESUMO: Hyptis marrubioides (Lamiaceae) é uma planta medicinal nativa do Cerrado brasileiro. Técnicas de propagação in vitro fazem uso de elicitores para alterar as vias metabólicas, afetando a produção de moléculas qualitativa e quantitativamente. Este trabalho teve como objetivo avaliar como os elicitores abióticos ácido salicílico (SA) e nitrato de prata (SN) nas concentrações de 30µM ou 60µM influenciam no crescimento de plântulas de Hyptis marrubioides por dois diferentes métodos de cultivo in vitro. O teor de rutina foi quantificado por CLAE-DAD. Em comparação com uma cultura não tratada, os extratos metanólicos de H. marrubioides cultivados em meio MS por 10 dias, seguidos de cultura em meio MS contendo SN (30µM) por 20 dias, apresentaram 1,28 vezes maior teor de rutina. Em um segundo experimento, as plântulas foram cultivadas em meio MS por 20 dias, e então o elicitor desejado foi adicionado à cultura e deixado em contato com o meio por três e seis dias. SA (30µM) forneceu os melhores resultados na produção de rutina, sendo 16,56 vezes maior do que o controle após seis dias. SN (30µM) aumentou o teor de rutina em 1,17 vezes. Nas duas concentrações avaliadas durante os experimentos de elicitação, nem SA nem SN alteraram os parâmetros de crescimento, como comprimento da parte aérea, número de folhas ou peso fresco e seco das plântulas de H. marrubioides cultivadas in vitro em relação ao controle. Com base nestes resultados, os elicitores abióticos SA e SN forneceram com sucesso Hyptis marrubioides in vitro com maior conteúdo de rutina.

Evaluation in vitro of the capacity of selection of the protetic canal with dentinary adhesive and different treatments of the dentinary remanescent / Avaliação in vitro da capacidade de selamento do canal protético com adesivo dentinário e diferentes tratamentos do remanescente dentinário

The evaluation of the in vitro evaluation, the ability to seal the prosthetic canal in bovine teeth with different treatments of the remaining dentin. Twenty-seven specifications were randomly assigned and divided into 6 groups: (A) without sealing of the prosthetic canal and filled with AH Plus; (B) without sealing of the prosthetic canal and filled with Endofill; (C) with sealing of the prosthetic canal and filled with AH Plus; (D) with sealing of the prosthetic canal and filled with Endofill; (E) with sealing of the prosthetic canal and filled with Endofill and dentin extension; (F) with sealing of the prosthetic canal and obturated with AH Plus and dental enlargement. After being immersed in silver nitrate solution, an evaluation of the sealing capacity of the prosthetic canal was performed by scanning electron microscopy by field emission. The results were experimental group A, B, C, D and E, degree of infiltration by means of silver nitrate; group E presented a higher infiltration score, different from group A, with statistically significant difference between groups E and F (p = 0.018). It was concluded that there was silver nitrate infiltration in the obturator material, to a greater or lesser degree, in five of the six experimental groups.

O objetivo deste estudo foi avaliar, in vitro, a capacidade de selamento do canal protético em dentes bovinos com diferentes tratamentos do remanescente dentinário. Vinte e sete espécimes foram distribuídos aleatoriamente e divididos em 6 grupos: (A) sem selamento do canal protético e obturadas com cimento AH Plus; (B) sem selamento do canal protético e obturadas com Endofill; (C) com selamento do canal protético e obturadas com AH Plus; (D) com selamento do canal protético e obturadas com Endofill; (E) com selamento do canal protético e obturadas com Endofill e alargamento dentinário; (F) com selamento do canal protético e obturadas com AH Plus e alargamento dentinário. Após serem imersos em solução de nitrato de prata, foi realizada a avaliação da capacidade de selamento do canal protético através da microscopia eletrônica de varredura por emissão de campo. Os resultados mostraram que os grupos experimentais A, B, C, D e E apresentaram grau de infiltração por meio do nitrato de prata; o grupo E apresentou maior escore de infiltração, diferente dos grupo A com o menor escore e o grupo F, que não apresentou infiltração quando comparado ao grupo E. Houve diferença estatisticamente significante apenas entre os grupos E e F (p=0,018). Concluiu-se que houve infiltração de nitrato de prata no material obturador, em maior ou menor grau, em cinco dos seis grupos experimentais.

Soluções de óxido de zinco e de nitrato de prata como alternativa para antissepsia de tetos de bovinos (teste in situ) / Solutions of Zinc oxide and silver nitrate as an alternative to bovine udder antisepsis

Este trabalho teve como objetivo analisar o potencial das soluções de óxido de zinco e de nitrato de prata como alternativa para antissepsia de tetos de bovinos (in situ), considerando a escassez de literatura a respeito do uso destes compostos na profilaxia de mastites. Primeiramente foram realizados ensaios de microdiluição e CBM (Concentração Bactericida Mínima) a fim de verificar a atividade antimicrobiana das soluções de óxido de zinco e de nitrato de prata sobre 30 isolados de Staphylococcus spp., obtidos de casos de mastite. Todos os isolados apresentaram sensibilidade às duas soluções testadas, mas a solução de nitrato de prata apresentou menores valores de CBMs (3,05 a 97,65 µg/mL), quando comparada as CBMs da solução de óxido de zinco (97,65 a 6.250 µg/mL). Posteriormente, foram conduzidos os ensaios de antissepsia dos tetos (in situ) através da imersão dos mesmos nas soluções antimicrobianas testadas. Para isso, foram utilizados 40 tetos (n=40) oriundos de vacas abatidas, os quais foram divididos em 4 grupos de 10 tetos, destinados ao teste das solução de óxido de zinco a 3% (30mg/mL), de nitrato e prata 1% (10mg/mL) e seus respectivos controles. Os tetos foram submetidos à contagem de microrganismos mesófilos na superfície dos tetos (UFC/cm2) antes e após a imersão nas soluções testadas. Como resultado, todas as soluções (de óxido de zinco e de nitrato de prata) apresentaram significativa redução de UFC/cm2 até aos 60 minutos após a imersão (M 0' a M 60'). A solução de nitrato de prata apresentou redução de UFC/cm2 significativamente maior, quando comparado ao grupo testado com solução de óxido de zinco. Tais resultados validam o potencial das soluções de óxido de zinco e nitrato de prata para utilização no pré e pós-dipping em vacas leiteiras.(AU)

This study aimed to analyze the zinc oxide and silver nitrate solutions as an alternative antiseptic for cow teats (in situ), considering the lack in the literature about these compounds uses in mastitis prophylaxis. First microdilution tests med and MBC (Minimal Bactericidal Concentration were perfor in order to determine the antimicrobial activity of zinc oxide and silver nitrate solutions over 30 Staphylococcus spp. isolates, obtained from cows with mastitis. All strains tested showed sensitivity to both solutions, but the silver nitrate solution had lower MBC values (3,05 to 97,65ug/ml) compared with zinc oxide solution MBCs (97,65 to 6,250 ug/ml). Subsequently, the antiseptic teat tests were conducted (in situ) by immersing the teats in same antimicrobial solutions tested in the first experiment. Therefore, 40 teats were used (n = 40) originating from slaughtered cows were divided into 4 groups of 10 teats, to test test zinc oxide 3% (30mg/mL) and silver nitrate 1% (10mg/mL) solutions and their respective controls. The teats were submitted to mesophilic count on the teat surface (CFU/cm2) before and after immersion in tested solutions. As a result, all the solutions (zinc oxide and silver nitrate) had a significant reduction in CFU/cm2 until 60 minutes after immersion (M 0' to 60 M'). Silver nitrate solution showed a reduction of CFU/cm2 significantly higher compared to the group treated with zinc oxide solution. These results validate the potential use of zinc oxide and silver nitrate solutions in dairy cows as a pre- and post-dipping antiseptic.(AU)

Soluções de óxido de zinco e de nitrato de prata como alternativa para antissepsia de tetos de bovinos (teste in situ) / Solutions of Zinc oxide and silver nitrate as an alternative to bovine udder antisepsis

Este trabalho teve como objetivo analisar o potencial das soluções de óxido de zinco e de nitrato de prata como alternativa para antissepsia de tetos de bovinos (in situ), considerando a escassez de literatura a respeito do uso destes compostos na profilaxia de mastites. Primeiramente foram realizados ensaios de microdiluição e CBM (Concentração Bactericida Mínima) a fim de verificar a atividade antimicrobiana das soluções de óxido de zinco e de nitrato de prata sobre 30 isolados de Staphylococcus spp., obtidos de casos de mastite. Todos os isolados apresentaram sensibilidade às duas soluções testadas, mas a solução de nitrato de prata apresentou menores valores de CBMs (3,05 a 97,65 µg/mL), quando comparada as CBMs da solução de óxido de zinco (97,65 a 6.250 µg/mL). Posteriormente, foram conduzidos os ensaios de antissepsia dos tetos (in situ) através da imersão dos mesmos nas soluções antimicrobianas testadas. Para isso, foram utilizados 40 tetos (n=40) oriundos de vacas abatidas, os quais foram divididos em 4 grupos de 10 tetos, destinados ao teste das solução de óxido de zinco a 3% (30mg/mL), de nitrato e prata 1% (10mg/mL) e seus respectivos controles. Os tetos foram submetidos à contagem de microrganismos mesófilos na superfície dos tetos (UFC/cm2) antes e após a imersão nas soluções testadas. Como resultado, todas as soluções (de óxido de zinco e de nitrato de prata) apresentaram significativa redução de UFC/cm2 até aos 60 minutos após a imersão (M 0' a M 60'). A solução de nitrato de prata apresentou redução de UFC/cm2 significativamente maior, quando comparado ao grupo testado com solução de óxido de zinco. Tais resultados validam o potencial das soluções de óxido de zinco e nitrato de prata para utilização no pré e pós-dipping em vacas leiteiras.(AU)

This study aimed to analyze the zinc oxide and silver nitrate solutions as an alternative antiseptic for cow teats (in situ), considering the lack in the literature about these compounds uses in mastitis prophylaxis. First microdilution tests med and MBC (Minimal Bactericidal Concentration were perfor in order to determine the antimicrobial activity of zinc oxide and silver nitrate solutions over 30 Staphylococcus spp. isolates, obtained from cows with mastitis. All strains tested showed sensitivity to both solutions, but the silver nitrate solution had lower MBC values (3,05 to 97,65ug/ml) compared with zinc oxide solution MBCs (97,65 to 6,250 ug/ml). Subsequently, the antiseptic teat tests were conducted (in situ) by immersing the teats in same antimicrobial solutions tested in the first experiment. Therefore, 40 teats were used (n = 40) originating from slaughtered cows were divided into 4 groups of 10 teats, to test test zinc oxide 3% (30mg/mL) and silver nitrate 1% (10mg/mL) solutions and their respective controls. The teats were submitted to mesophilic count on the teat surface (CFU/cm2) before and after immersion in tested solutions. As a result, all the solutions (zinc oxide and silver nitrate) had a significant reduction in CFU/cm2 until 60 minutes after immersion (M 0' to 60 M'). Silver nitrate solution showed a reduction of CFU/cm2 significantly higher compared to the group treated with zinc oxide solution. These results validate the potential use of zinc oxide and silver nitrate solutions in dairy cows as a pre- and post-dipping antiseptic.(AU)

Effect of thermocycling to adhesive interface of enamel analyzed by polarized light microscopy / Análise do efeito da termociclagem na interface adesivo do esmalte analisada por microscopia de luz polarizada

Objetivo: A desmineralização menos agressiva do esmalte por sistemas adesivos autocondicionantes resulta em maior descoloração, infiltração marginal e falhas em suas restaurações. Deste modo, este estudo teve como objetivo avaliar a microinfiltração por meio da técnica da infiltração de nitrato de prata e formação de tags na interface esmalte/adesivo. Material e Método: Duzentos fragmentos de esmalte foram divididos aleatoriamente em 10 grupos (n = 10) de acordo com o sistema adesivo (Single Bond Adper Plus- (SB), Clearfil Tri-S Bond- (CF), ou Scotchbond Universal- (SBU)) e a superfície do esmalte (lixada (GE), não lixada- (UE), condicionada com ácido fosfórico 37%- (PHA), ou sem condicionamento): (SB-UE), (SB-GE), (CF-UE), (CFGE), (CF-ue/pha), (CF-ge/pha), (SBU-ue), (SBU-ge), (SBU-ue/pha) e (SBU-ge/pha). Metade das amostras restauradas foram submetidas a 20.000 ciclos térmicos. Quatro fatias de 1,0 mm/amostra foram obtidas para avaliar a formação de tags e infiltração de nitrato de prata. Todas as amostras foram examinadas com microscopia de luz polarizada e a percentagem de infiltração foi quantificada. Resultados: Não foram observadas interações entre os três fatores. O sistema adesivo e envelhecimento exibiram uma interação. Foram encontradas diferenças significativas somente após termociclagem: os grupos SB e SBU condicionados apresentaram menor porcentagem de infiltração comparados aos outros grupos, independente do tipo de esmalte. Quanto à analise qualitativa, o comprimento dos tags após condicionamento ácido foi maior para o GE comparado ao UE, independente do sistema adesivo. Conclusão: A aplicação dos sistemas adesivos na técnica autocondicionante mostrou uma formação de tags significativamente menor comparada à técnica convencional. O condicionamento ácido do esmalte previamente aos sistemas adesivos multi-mode é fundamental para reduzir o grau de infiltração na interface adesiva após envelhecimento.

Objective: The less aggressive demineralization of enamel by self-etching systems results in greater staining, marginal leakage, and failure in their restorations, so this study aimed to assess the silver nitrate infiltration and tag formation of the enamel/adhesive interface. Material and Methods: Two hundred enamel fragments were randomly assigned into 10 groups according to the adhesive system (Single Bond Adper Plus- (SB), Clearfil Tri-S Bond-(CF), or Scotchbond Universal-(SBU)) and enamel surface (ground- (ge), unground-(ue), phosphoric acid etching- (pha), or none) (n=10): (SB-ue), (SB-ge), (CF-ue), (CF-ge), (CF-ue/pha), (CF-ge/pha), (SBU-ue), (SBU-ge), (SBU-ue/pha), and (SBUge/pha). Half of the restored samples were submitted to thermocycling. Four slices of 1.0mm/ sample were obtained to evaluate tag formation and silver nitrate infiltration. All of the specimens were examined with Polarized Light Microscopy, and the percentage of infiltration was quantified. Results: No interactions were found among the three factors. The adhesive and aging exhibited an interaction. Significant differences were found only after thermocycling: the SB and SBU-etched groups had decreased infiltration compared with the other groups. The tag length after etching was higher for ge compared with ue, regardless of the adhesive system. Conclusion: The selfetching techniques resulted in significantly less tag formation compared with the conventional technique. The acid pre-etching of enamel with the multi-mode adhesive was fundamental for reducing the degree of infiltration of the adhesive interface after aging