Comparison of different methods for total RNA extraction from sclerotia of Rhizoctonia solani

Background Rhizoctonia solani (teleomorph: Thanatephorus cucumeris) is one of the most important pathogens of rice (Oryza sativa L.) that causes severe yield losses in all rice-growing regions. Sclerotia, formed from the aggregation of hyphae, are important structures in the life cycles of R. solani and contain a large quantity of polysaccharides, lipids, proteins and pigments. In order to extract high-quality total RNA from the sclerotia of R. solani, five methods, including E.Z.N.A.™ Fungal RNA Kit, sodium dodecyl sulfate (SDS)-sodium borate, SDS-polyvinylpyrrolidone (PVP), guanidinium thiocyanate (GTC) and modified Trizol, were compared in this study. Results The electrophoresis results showed that it failed to extract total RNA from the sclerotia using modified Trizol method, whereas it could extract total RNA from the sclerotia using other four methods. Further experiments confirmed that the total RNA extracted using SDS-sodium borate, SDS-PVP and E.Z.N.A.™ Fungal RNA Kit methods could be used for RT-PCR of the specific amplification of GAPDH gene fragments, and that extracted using GTC method did not fulfill the requirement for above-mentioned RT-PCR experiment. Conclusion It is concluded that SDS-sodium borate and SDS-PVP methods were the better ones for the extraction of high-quality total RNA that could be used for future gene cloning and expression studies, whereas E.Z.N.A.™ Fungal RNA Kit was not taken into consideration when deal with a large quantity of samples because it is expensive and relatively low yield.

Effect of denture cleansers on metal ion release and surface roughness of denture base materials

Chemical disinfectants are usually associated with mechanical methods to remove stains and reduce biofilm formation. This study evaluated the effect of disinfectants on release of metal ions and surface roughness of commercially pure titanium, metal alloys, and heat-polymerized acrylic resin, simulating 180 immersion trials. Disk-shaped specimens were fabricated with commercially pure titanium (Tritan), nickel-chromium-molybdenum-titanium (Vi-Star), nickel-chromium (Fit Cast-SB Plus), and nickel-chromium-beryllium (Fit Cast-V) alloys. Each cast disk was invested in the flasks, incorporating the metal disk to the heat-polymerized acrylic resin. The specimens (n=5) were immersed in these solutions: sodium hypochlorite 0.05%, Periogard, Cepacol, Corega Tabs, Medical Interporous, and Polident. Deionized water was used as a control. The quantitative analysis of metal ion release was performed using inductively coupled plasma mass spectrometry (ELAN DRC II). A surface analyzer (Surftest SJ-201P) was used to measure the surface roughness (µm). Data were recorded before and after the immersions and evaluated by two-way ANOVA and Tukey's test (α=0.05). The nickel release proved most significant with the Vi-Star and Fit Cast-V alloys after immersion in Medical Interporous. There was a significant difference in surface roughness of the resin (p=0.011) after immersion. Cepacol caused significantly higher resin roughness. The immersion products had no influence on metal roughness (p=0.388). It could be concluded that the tested alloys can be considered safe for removable denture fabrication, but disinfectant solutions as Cepacol and Medical Interporous tablet for daily denture immersion should be used with caution because it caused greater resin surface roughness and greater ion release, respectively.

Desinfetantes químicos são normalmente associados a métodos mecânicos para remover manchas e reduzir a formação do biofilme. Este estudo avaliou o efeito de desinfetantes na liberação de íons metálicos e na rugosidade superficial do titânio comercialmente puro, ligas metálicas e resina acrílica termopolimerizável, simulando 180 ensaios de imersões. Espécimes em formato de discos foram confeccionados com titânio comercialmente puro (Tritan), liga de níquel-cromo-molibdênio-titânio (Vi-Star), liga de níquel-cromo (Fit Cast-SB Plus) e liga de níquel-cromo-berílio (Fit Cast-V). Os espécimes (n=5) foram imersos nestas soluções: hipoclorito de sódio a 0,05%, Periogard, Cepacol, Corega Tabs, Medical Interporous e Polident. Como controle, foi utilizada a água deionizada. A análise quantitativa de liberação de íons metálicos foi realizada por meio de espectrometria de massa com plasma indutivamente acoplado (ELAN DRC II). O rugosímetro (Surftest SJ-201P) foi utilizado para medir a rugosidade superficial (µm). Os dados foram registrados antes e depois das imersões e avaliados por ANOVA com dois fatores e teste de Tukey (α=0,05). A liberação de níquel provou ser mais expressiva nas ligas Vi-Star e Fit Cast-V após a imersão em Medical Interporous. Houve diferença significante na rugosidade superficial da resina (p=0,011) após a imersão. O Cepacol causou maior rugosidade superficial de forma significativa. Os produtos de imersão não influenciaram nos resultados da rugosidade do metal (p=0,388). Pode-se concluir que as ligas metálicas testadas podem ser consideradas seguras para a fabricação de próteses removíveis, mas as soluções desinfetantes como o Cepacol e a pastilha Medical Interporous para a imersão diária da prótese devem ser utilizados com cautela, pois causaram maior rugosidade superficial da resina e maior liberação de íons, respectivamente.

Effect of different cleansers on the surface of removable partial denture

Removable partial dentures (RPD) demand specific hygienic cleaning and the combination of brushing with immersion in chemical solutions has been the most recommended method for control of biofilm. However, the effect of the cleansers on metallic components has not been widely investigated. This study evaluated the effect of different cleansers on the surface of RPD. Five disc specimens (12 mm x 3 mm metallic disc centered in a 38 x 18 x 4 mm mould filled with resin) were obtained for each experimental situation: 6 solutions [Periogard (PE), Cepacol (CE), Corega Tabs (CT), Medical Interporous (MI), Polident (PO), 0.05 percent sodium hypochlorite (NaOCl), and distilled water (DW) control] and 2 Co-Cr alloys [DeguDent (DD) and VeraPDI (VPDI)] were used for each experimental situation. A 180-day immersion was simulated and the measurements of roughness (Ra, µm) of metal and resin were analyzed using 2-way ANOVA and Tukey’s test. The surface changes and tarnishes were examined with a scanning electronic microscopy (SEM). In addition, energy dispersive x-ray spectrometry (EDS) analysis was carried out at representative areas. Visually, NaOCl and MI specimens presented surface tarnishes. The roughness of materials was not affected by the solutions (p>0.05). SEM images showed that NaOCl and MI provided surface changes. EDS analysis revealed the presence of oxygen for specimens in contact with both MI and NaOCl solutions, which might suggest that the two solutions promoted the oxidation of the surfaces, thus leading to spot corrosion. Within the limitations of this study, it may be concluded that the NaOCl and MI may not be suitable for cleaning of RPD.

As próteses parciais removíveis (PPR) exigem higienização específica e a associação da escovação com imersão em soluções químicas tem sido o método mais recomendado para controle do biofilme. Entretanto, os efeitos destas soluções não são amplamente reportados em componentes metálicos. Este estudo avaliou o efeito de diferentes agentes de higienização na superfície dos componentes de uma PPR. Foram confeccionados 5 espécimes (disco metálico de 12 x 3 mm centralizado em uma tira de resina com 38 x 18 x 4 mm) para cada situação experimental: 6 soluções [Periogard (PE), Cepacol (CE), Corega Tabs (CT), Medical Interporous (MI), Polident (PO), hipoclorito de sódio 0,05 por cento (HS) e água destilada (AD) como controle)] e 2 ligas de cobalto-cromo [DeguDent (DD) e Vera PDI (VPDI)] foram utilizadas para cada situação experimental. Foram simuladas imersões de 180 dias. As aferições de rugosidade (Ra, μm) tanto em porção metálica quanto em resina acrílica termopolimerizável foram submetidos ao ANOVA e ao teste de Tukey. As alterações superficiais e manchas foram examinadas por meio de microscopia eletrônica de varredura (MEV). Áreas de interesse foram submetidas à espectrometria por energia dispersiva por raios X (EDS). Visualmente, puderam ser verificadas manchas nas superfícies metálicas quando utilizados HS e MI. A rugosidade dos materiais não foi afetada pelas soluções (p>0,05). As fotomicrografias evidenciaram que HS e MI ocasionaram alterações superficiais. As análises de EDS revelaram a presença de oxigênio nos grupos HS e MI, o que pode sugerir que estas duas soluções causaram oxidação das superfícies, provocando pontos de corrosão. Dentre as limitações do presente estudo, pode-se concluir que estas soluções não são apropriadas para a higienização das PPR.

Evaluation of alternative methods for the disinfection of toothbrushes

The aim of this study was to evaluate alternative methods for the disinfection of toothbrushes considering that most of the previously proposed methods are expensive and cannot be easily implemented. Two-hundred toothbrushes with standardized dimensions and bristles were included in the study. The toothbrushes were divided into 20 experimental groups (n = 10), according to microorganism considered and chemical agent used. The toothbrushes were contaminated in vitro by standardized suspensions of Streptococcus mutans, Streptococcus pyogenes, Staphylococcus aureus or Candida albicans. The following disinfectants were tested: 0.12 percent chlorhexidine digluconate, 50 percent white vinegar, a triclosan-containing dentifrice solution, and a perborate-based tablet solution. The disinfection method was immersion in the disinfectant for 10 min. After the disinfection procedure, the number of remaining microbial cells was evaluated. The values of cfu/toothbrush of each group of microorganism after disinfection were compared by Kruskal-Wallis ANOVA and Dunn's test for multiple comparisons (5 percent). The chlorhexidine digluconate solution was the most effective disinfectant. The triclosan-based dentifrice solution promoted a significant reduction of all microorganisms' counts in relation to the control group. As to the disinfection with 50 percent vinegar, a significant reduction was observed for all the microorganisms, except for C. albicans. The sodium perborate solution was the less effective against the tested microorganisms. Solutions based on triclosan-containing dentifrice may be considered effective, nontoxic, cost-effective, and an easily applicable alternative for the disinfection of toothbrushes. The vinegar solution reduced the presence of S. aureus, S. mutans and S. pyogenes on toothbrushes.

Phosphate-borate ester preparation, structural elucidation and application as hydraulic brake fluid base

Hydraulic brake fluid is an area for considerable research effort. These fluids must possess high chemical and thermal stability, low corrosivity and high degree of water tolerance. These fluids are formulated from base, diluent and additives. Different structures, based on tri- and bis-borate as well as phosphate esters of hydraulic brake fluids are reported in the literature. [1] The incompatibility, deterioration, easy oxidation and instability are disadvantages of some of the above mentioned compounds. Their use is handicapped by the fact that a wide range of suitable properties cannot be attained.[1,2] Accordingly, the idea for the preparation ol triphosphate-borate ester was originated. We presumed that they may exhibit excellent properties of high degree of compatibility with other hydraulic brake fluids, high stability and are useful under a wide variety of climatic and operational conditions. We also predict that they meet the standard specifications