Toluene permeabilization differentially affects F- and P-type ATPase activities present in the plasma membrane of Streptococcus mutans

Streptococcus mutans membrane-bound P- and F-type ATPases are responsible for H+ extrusion from the cytoplasm thus keeping intracellular pH appropriate for cell metabolism. Toluene-permeabilized bacterial cells have long been used to study total membrane-bound ATPase activity, and to compare the properties of ATPase in situ with those in membrane-rich fractions. The aim of the present research was to determine if toluene permeabilization can significantly modify the activity of membrane-bound ATPase of both F-type and P-type. ATPase activity was assayed discontinuously by measuring phosphate release from ATP as substrate. Treatment of S. mutans membrane fractions with toluene reduced total ATPase activity by approximately 80 percent and did not allow differentiation between F- and P-type ATPase activities by use of the standard inhibitors vanadate (3 µM) and oligomycin (4 µg/mL). Transmission electron microscopy shows that, after S. mutans cells permeabilization with toluene, bacterial cell wall and plasma membrane are severely injured, causing cytoplasmic leakage. As a consequence, loss of cell viability and disruption of H+ extrusion were observed. These data suggest that treatment of S. mutans with toluene is an efficient method for cell disruption, but care should be taken in the interpretation of ATPase activity when toluene-permeabilized cells are used, because results may not reflect the real P- and F-type ATPase activities present in intact cell membranes. The mild conditions used for the preparation of membrane fractions may be more suitable to study specific ATPase activity in the presence of biological agents, since this method preserves ATPase selectivity for standard inhibitors.

Potencial tóxico dos alginatos para uso odontológico / Potencial toxicity of dental alginates

O alginato ou hidrocolóide irreversível é um dos materiais de moldagem mais aceitos e utilizados na Odontologia. Muitas substâncias como zinco, cádmio, silicato de chumbo e fluoretos foram adicionadas em algumas marcas de alginatos, com o objetivo de melhorar suas propriedades físicas, químicas, mecânicas e se tornaram causa de preocupação no que se refere à toxicidade desses materiais. Em algumas marcas de alginatos relatou-se a presença de fluoretos, cádmio, silicatos de chumbo e zinco potencialmente tóxicos, isoladamente ou em conjunto, consequentemente, cuidados especiais devem ser tomados na preparação desses materiais. É necessário que haja um controle contínuo de metais e substâncias potencialmente tóxicas nos alginatos para se evitar a contaminação dos profissionais da área odontológica e pacientes. Nesta revisão analisou-se o potencial tóxico de alginatos usados em odontologia.

Análise quantitativa de fluoretos nos alginatos para uso odontológico / Quantitative analysis of fluorides in alginates for dental use

Este trabalho teve como proposta analisar quantitativamente o conteúdo de fluoretos nos alginatos para uso odontológico e a liberação de fluoretos de moldes desses alginatos em água milliQ,saliva artificial e ácido clorídrico 0,1 mol/l.Foram investigadas sete marcas de alginatos disponíveis comercialmente no Brasil, sendo analisados dois lotes de cada material.As concentrações de fluoretos nas diferentes amostras foram determinadas por potenciometria direta, utilizando o eletrodo seletivo combinado de fluoreto.Os materiais que apresentaram maiores concentrações médias de fluoreto total foram Hydrogum (7052,87μg/g), Jeltrate Plus (6519,68μg/g) e Orthoprint (6218,18μg/g).Apenas os materiais das marcas Hydrogum e Jeltrate apresentaram diferenças nas concentrações de fluoretos entre os lotes um e dois.Os materiais apresentaram diferenças na liberação de fluoretos dos moldes, cujas maiores concentrações médias foram liberadas pelas marcas Hydrogum e Orthoprint.O meio influenciou na liberação de fluoreto, sendo que na saliva foi menor que na água e nesta foi inferior ao ácido.Os moldes dos materiais que mais liberaram fluoretos nos três meios (saliva, água e ácido) foram os do Hydrogum e Orthoprint.Considerando que as concentrações de fluoretos encontradas nos alginatos são altas e que existem diferentes fontes de exposição aos fluoretos, há necessidade de constante monitoramento dos alginatos para uso odontológico

Quantitative analysis of potentially toxic metals in alginates for dental use

Alginate is one de materials most employed in practice to make dental impressions. Substances like zinc, cadmium and lead silicate, which are included in several alginate brands with the aim of improving their physical, chemical and mechanical properties, are a source of serious concern regards their toxicity. The most serious chronic effect of oral exposure to cadmium is renal toxicity. Assimilation of lead has deleterious effects on the gastrointestinal tract, hematopoietic system, cardiovascular system, central and peripheral nervous systems, kidneys, immune system, and reproductive system. Chronic oral exposures to zinc have resulted in hypochromic and microcyte anemia in some individuals. The aim of the present study was to measure the cadmium, lead and zinc contents of seven brands of alginate for dental use on sale in Brazil. The samples were weighed and placed in the Teflon cups of a closed-system microwave oven. Aqua regia (4 mL concentrated HCl:HNO3, 3:1 v/v) and hydrofluoric acid (2mL concentrated HF) were added to the samples, which were then subjected to heating. The samples were then cooled to room temperature and diluted to 25 mL in deionized water in a volumetric glass flask. The samples were diluted in duplicate and analyzed against a reagent blank. The analyses were performed in an atomic absorption flame spectrophotometer. Neither lead nor cadmium was detected. Zinc contents ranged from 0.001% to 1.36% by weight. The alginates exhibited low contents of the metals under study and gave no cause for concern regarding toxicity; even so, it is advisable to monitor potentially toxic materials continually and to analyze their plasmatic levels in the professionals working with them