RESUMO
The investigation was carried out by evaluating the microbiological characteristics of the water before and after treatment with Er:YAG laser and turbine. The study was carried out in 2 dental surgeries. In both cases the laser and dental units were served by two independent circuits, fed by the same potable tap water. Samples were taken from the water supplying and the water leaving the turbine and laser before and after treatment on the same patient. Total heterotrophic plate count was measured at 36 degrees C and at 22 degrees C, and the presence of Staphylococcus species and non-fermenting Gram negative bacteria was investigated. Bacterial contamination was found within the circuit, especially in the laser device. Pseudomonas aeruginosa was detected in only 1 sample of supply water, in 11.1 % and in 19.4 % of the samples from the turbine and the laser respectively. No evidence of Staphylococcus aureus was found. The contamination of supply water was low, whereas that of the water leaving the handpieces of the 2 devices was high, especially in the laser. Attention should be paid to the control of the water leaving laser devices, given the increasingly wide use of such instruments in dental treatment exposed to risk of infection.
Assuntos
Equipamentos Odontológicos de Alta Rotação/microbiologia , Desinfecção/métodos , Contaminação de Equipamentos/prevenção & controle , Lasers , Purificação da Água/métodos , Descontaminação/métodos , Humanos , Controle de Infecções/instrumentação , Itália , Estatísticas não Paramétricas , Microbiologia da ÁguaRESUMO
The mechanisms that determine how folding attempts are interrupted to target folding-incompetent proteins for endoplasmic reticulum-associated degradation (ERAD) are poorly defined. Here the alpha-mannosidase I-like protein EDEM was shown to extract misfolded glycoproteins, but not glycoproteins undergoing productive folding, from the calnexin cycle. EDEM overexpression resulted in faster release of folding-incompetent proteins from the calnexin cycle and earlier onset of degradation, whereas EDEM down-regulation prolonged folding attempts and delayed ERAD. Up-regulation of EDEM during ER stress may promote cell recovery by clearing the calnexin cycle and by accelerating ERAD of terminally misfolded polypeptides.
Assuntos
Ácido Aspártico Endopeptidases/metabolismo , Calnexina/metabolismo , Retículo Endoplasmático/metabolismo , Glicoproteínas/metabolismo , Proteínas de Membrana/metabolismo , Ácido Aspártico Endopeptidases/química , Linhagem Celular , Regulação para Baixo , Eletroforese em Gel de Poliacrilamida , Glicoproteínas/química , Glicosilação , Humanos , Cinética , Peso Molecular , Polissacarídeos/metabolismo , Conformação Proteica , Dobramento de Proteína , Interferência de RNA , Transfecção , Regulação para CimaRESUMO
OBJECTIVE: Iron deficiency is estimated to affect about 30% of the world population. Iron supplementation in the form of tablets and food fortification has not been successful in developing countries, and iron deficiency is still the most important deficiency related to malnutrition. Here we present experiments that aim to increase the iron content in rice endosperm and to improve its absorption in the human intestine by means of genetic engineering. METHODS: We first introduced a ferritin gene from Phaseolus vulgaris into rice grains, increasing their iron content up to twofold. To increase iron bioavailability, we introduced a thermo-tolerant phytase from Aspergillus fumigatus into the rice endosperm. In addition, as cysteine peptides are considered major enhancers of iron absorption, we over-expressed the endogenous cysteine-rich metallothionein-like protein. RESULTS: The content of cysteine residues increased about sevenfold and the phytase level in the grains about one hundred and thirtyfold, giving a phytase activity sufficient to completely degrade phytic acid in a simulated digestion experiment. CONCLUSIONS: This rice, with higher iron content, rich in phytase and cysteine-peptide has a great potential to substantially improve iron nutrition in those populations where iron deficiency is so widely spread.
Assuntos
Anemia Ferropriva/dietoterapia , Anemia Ferropriva/metabolismo , Ferro da Dieta/metabolismo , Oryza/genética , Oryza/metabolismo , Plantas Geneticamente Modificadas/metabolismo , Sementes/genética , Sementes/metabolismo , 6-Fitase/genética , Cromatografia Líquida de Alta Pressão , Ferritinas/genética , Humanos , Metaloproteínas/genética , Proteínas de Plantas/genética , Espectrofotometria AtômicaRESUMO
To obtain a functioning provitamin A (beta-carotene) biosynthetic pathway in rice endosperm, we introduced in a single, combined transformation effort the cDNA coding for phytoene synthase (psy) and lycopene beta-cyclase (beta-lcy) both from Narcissus pseudonarcissus and both under the control of the endosperm-specific glutelin promoter together with a bacterial phytoene desaturase (crtI, from Erwinia uredovora under constitutive 35S promoter control). This combination covers the requirements for beta-carotene synthesis and, as hoped, yellow beta-carotene-bearing rice endosperm was obtained in the T(0)-generation. Additional experiments revealed that the presence of beta-lcy was not necessary, because psy and crtI alone were able to drive beta-carotene synthesis as well as the formation of further downstream xanthophylls. Plausible explanations for this finding are that these downstream enzymes are constitutively expressed in rice endosperm or are induced by the transformation, e.g., by enzymatically formed products. Results using N. pseudonarcissus as a model system led to the development of a hypothesis, our present working model, that trans-lycopene or a trans-lycopene derivative acts as an inductor in a kind of feedback mechanism stimulating endogenous carotenogenic genes. Various institutional arrangements for disseminating Golden Rice to research institutes in developing countries also are discussed.