Manual and Rotary Instrumentation Ability to Reduce Enterococcus faecalis Associated with Photodynamic Therapy in Deciduous Molars

This aim of this study was to assess the ability of manual or rotary instrumentation associated with photodynamic therapy (PDT) to reduce Enterococcus faecalis using three combinations of light/photosensitizers: toluidine blue O/laser, fuchsin/halogen light and fuchsin/LED. Twenty deciduous molars were selected and contaminated with Enterococcus faecalis (McFarland 0.5 scale). Working length determination was performed by visual method. The teeth were randomly divided into two groups: G1 (n=10): manual instrumentation (Kerr-type files) and G2 (n=10): rotary instrumentation (ProTaper system). The bacteria were collected three times using sterile paper cones compatible with the anatomic diameter of the root canal for 30 s before and after instrumentation and after PDT. The samples were diluted in peptone water, seeded on blood agar plates and incubated in an oven at 37 °C for colony-forming units counting. The decrease of E. faecalis counts after instrumentation and after PDT was compared using the Wilcoxon test, t-test and Kruskal Wallis test. A significant reduction of E. faecalis occurred after manual and rotary instrumentation and after PDT using the three combinations of light/photosensitizer (p<0.05). It may be concluded that both rotary and manual instrumentation reduced E. faecalis. Fuchsin with halogen light or LED irradiation and toluidine blue O with laser irradiation can be used to reduce E. faecalis in root canals of primary molars. PDT can be used as an adjuvant to conventional endodontic treatment.

O objetivo do presente estudo foi avaliar a redução de Enterococcus faecalis após instrumentação manual ou rotatória associada à terapia fotodinâmica (PDT) utilizando 3 combinações luz/fotossensibilizante: azul de toluidina O/laser, fucsina/luz halógena e fucsina/LED. Foram selecionados 20 molares decíduos que foram contaminados com Enterococcus faecalis (escala 0,5 de McFarland). A odontometria foi feita através do método visual. Os dentes foram divididos aleatoriamente em dois grupos: G1 (n=10): instrumentação manual (limas tipo Kerr) e G2 (n=10): instrumentação rotatória (sistema ProTaper). Foram realizadas coletas com cone de papel estéril compatível com o diâmetro anatômico do canal durante 30 s antes e após a instrumentação e a PDT. As amostras foram diluídas em água peptonada, semeadas em placas de agar-sangue e incubadas em estufa a 37 °C para contagem das unidades formadoras de colônias. As comparações antes da redução de E. faecalis após a instrumentação e após a realização da PDT foram realizadas pelo teste de Wilcoxon, teste t e Kruskal Wallis. Houve redução significante de E. faecalis após a instrumentação manual ou rotatória e após realização da PDT com as três combinações de luz/fotossensibilizante (p<0,05). Pode-se concluir que a instrumentação rotatória e manual acarretou a redução de E. faecalis. A fucsina irradiada com luz halógena ou led e o azul de toluidina irradiado com laser podem ser utilizados para redução de E. faecalis do sistema de canais radiculares de molares decíduos. A terapia fotodinâmica pode ser utilizada como coadjuvante ao tratamento endodôntico convencional.

Efeito das vastatinas sobre a hidrólise hepática do colesterol esterificado, em células de hepatoblastoma humano HEP G2 / Effects of vastatins in the hepatic hidrolase of esterified cholesterol in human HEP G2 hepatoblastoma cells

O presente trabalho se valeu de células de hepatoma humano Hep G2 para estudar se existe algum efeito das vastatinas sobre a atividade das enzimas que governam a hidrólise hepática do coleterol: colesterol éster hidrolases lisossomal, citosólica e microssomal. As enzimas foram incubadas com ou sem amostras de vastatina, dissovidas em dimetilsulfóxido. A adiçao das vastatinas ofereceu distintos resultados, conforme a maneira que se apresentou o substrato às enzimas. No caso da colesterol éster hidrolase lisossomal, obteve-se inibiçao em substrato Hajjar, com sais biliares, e estimulaçao em substrato Brecher, desprovido de sais biliares. A hidrolase citosólica teve sua atividade inibida em substrato Hajjar e constante em substrato Brecher.

Subcellular distribution of oxygen free radical scavenging enzymes in goat ovary.

Oxygen free radical scavenging enzymes superoxide dismutase (SOD), peroxidase (POD) and catalase were heterogeneously distributed in goat ovary. Activities of SOD and catalase were predominantly located in cytosolic fractions compared to mitochondrial and microsomal fractions. Most of the peroxidase activity was observed in microsomal fractions with little activity in cytosolic and mitochondrial fractions. Higher activities of all these enzymes were in luteal phase as compared to follicular phase. Most of the activities of these enzymes in luteal phase were restricted to luteal cells while in follicular phase these were mainly present in the follicles of the ovary.

A brain soluble fraction inhibits synaptosomal membrane but not astrocyte ATPase activity

Since a brain soluble fraction (peak II) is known to be able to inhibit synaptosomal membrane Na+, K+-ATPase activity, here we attempted to compare its effect on cellular and subcellular brain components such as synaptosomal and astrocytic membranes, as well as mitochondrial preparations. Peak II highly inhibited total ATPase in synaptosomal membranes but failed to modify enzyme activity in astrocytic and mitochondrial preparations. Findings suggest cellular and subcellular specificity of peak II on brain ATPase activity.

Studies on cinnamic acid-4-hydroxylase from wounded potato tissue: isolation and characterization of cinnamic acid-4-hydroxylase activation factor.

Cinnamic acid-4-hydroxylase activation factor has been found to be located in the supernatant fraction of wounded potato tissue homogenate in phosphate buffer. The factor has been purified to homogeneity as judged by SDS polyacrylamide gel electrophoresis, by heat treatment on boiling water-bath for 7.5 min followed by dialysis with cut off limit of 8 kDa and final separation by gel filtration on Sephadex G-50 column. Gel filtration resolved this into three active fractions of molecular mass 12500, 10000 and 8500 Da conjugated to a fluorescent compound and subsequently identified as a folate derivative. The amino acid analysis of polypeptide chains of these fractions revealed that the polypeptides were rich in glutamic and aspartic acids. The fluorescent moiety of the complex released from polypeptide chain of molecular weight 10000 by mild acid hydrolysis was able to support the growth of Lactobacillus casei ATCC 7469 which requires folic acid for its growth. On storage, this compound degraded into a number of fluorescent products identified as p-amino benzoic acid, p-amino benzoyl glutamic acid, pteroic acid and 6-methyl pterin indicating that the activation factor is a folic acid derivative conjugated with the polypeptide chain.

Subcellular distribution of superoxide dismutase and catalase in human malarial parasite Plasmodium vivax.

Endogenous superoxide dismutase (SOD) has not been found to be present in P. vivax, a human malarial parasite and therefore it adopts and concentrates SOD from the host cell erythrocytes. It is demonstrated here that this adopted SOD from the host gets localized in lysosomes (10 k and 100 k fractions) of the malarial merozoites. P. vivax parasites were also found to contain very low levels of catalase, presumably as a result of contamination or adoption from the host red cell materials. It is therefore suggested that P. vivax merozoites are deficient in enzymes which are protective against the reactive oxygen species.

Identification of a non-microsomal cinnamic acid 4-hydroxylase from potato tuber (S. tuberosum) and its partial purification.

The cytoplasmic localisation of cinnamic acid 4-hydroxylase (CA4H) has been shown by isolation and subcellular fractionation of the enzyme in Hepes buffer. The enzyme was purified by ammonium sulphate fractionation followed by AcA-34 molecular sieve chromatography. The enzyme existed as a high molecular mass which dissociated to a lower form on dilution on the column. The pH optimum, sulphydryl requirement, molecular and preliminary kinetic characteristics were investigated.

Characterization of amyloplastic phosphohexose isomerase from immature wheat (Triticum aestivum L.) endosperm.

Phosphohexose isomerase from amyloplasts of immature wheat endosperm was purified 133-fold. The enzyme had a molecular weight of 130 kDa and maximum activity at pH 8.6. It showed normal hyperbolic kinetics for both fructose-6-P and glucose-6-P with Km of 0.12 mM and 0.44 mM, respectively. pH had a great influence on Km for fructose-6-P. Using glucose-6-P as the substrate, the equilibrium was reached at 23% fructose-6-P and 77% glucose-6-P and an equilibrium constant of about 3.0. The delta F calculated from the apparent equilibrium constant was +742 cal.mol-1. The activation energy calculated from the Arrhenius plot was 7450 cal.mol-1. None of the sulphydryl reagents at 2.5 mM concentration inactivated the enzyme. The enzyme was competitively inhibited by 6-phosphogluconate, ribose-5-P and ribulose-5-P with Ki values of 0.18, 0.14, and 0.13 mM, respectively. The probable role of the enzyme in starch biosynthesis in amyloplasts is discussed.

Proteolytic modification of growth hormone by a rat kidney lysosomal protease

Se detectó en riñon de rata una proteasa que hidroliza la hormona de crecimiento bovina (bGH) con cierto grado de especificidad. La enzima tiene un pH óptimo de 5.0, requiere la presencia de tioles en el medio de incubación y no es inhibida por EDTA o Aprotinina. Por digestión enzimática controlada de la bGH se obtuvo un derivado de la hormona, con propiedades fisicoquímicas diferentes a las de la proteína nativa, que retenía la actividad promotora de crecimiento en ratas hipofisoprivas. Los resultados sugieren que en el riñón, así como sucedería en otros tejidos, se originan formas activas de las hormonas de crecimiento

Effects of synthetic and natural food colourants on subcellular fraction enzymes in albino rats

The effect of synthetic food colorants as well as natural food colorants on subcellular fraction enzymes involving the pentose phosphate pathway, was studied. The results revealed that glucose-6 phosphate dehydrogenase [G-6-P.D] and 6-phosphogluconate dehydrogenase [6-PG.D] activities in cytoplasm and mitochondria of brain, liver and kidney were increased by single dose and daily dosage for 3 weeks. This increase was greater by synthetic food colorants than by natural food colorants. Also, the activity of G-6-P.D. was more affected than the activity of 6-P-G.D. particularly in cytoplasm. The effect of synthetic food colorants was continued for one week after termination of treatment which lasted for 3 weeks. Meanwhile, these enzyme activities returned to the normal control values when treatment with natural food colorants stopped

Sub-cellular distribution of monoamine oxidase and monoamine dehydrogenase.

MAO and MADH activities in different sub-cellular fractions of rat brain have been studied. The sub-cellular distribution pattern of MAO and MADH was found to be same, mostly concentrated in the mitochondrial fraction, although activities were also present in microsome and nucleus.