Bud source, asepsis and benzylaminopurine (BAP) effect on yacon (Polymnia sonchifolia) micropropagation

The yacon (Polymnia sonchifolia) is used largely for the high fructan content of its tubers; consequently, it is a good alternative for diabetics. One of the more important restricting factors of the commercial production of yacon is its susceptibility to nematode attack. This, as well as germplasm bank maintenance, justifies the importance of in vitro propagation of this species. In this way, our work aimed to verify the best asepsis method for yacon for the in vitro establishment from the rhizophore and the axillary buds of the aerial parts, and the effect of benzylaminopurine (BAP) addition to the culture medium. The number of contaminated cultures, the occurrence of phenolic oxidation and the occurrence of a vitreous aspect, showed differences with bud source, immersion time for asepsis, and BAP use. The results contribute to establishing a yacon micro propagation procedure.

Purification and characterization of xylanases from Aspergillus giganteus

A strain of Aspergillus giganteus cultivated in a medium with xylan produced two xylanases (xylanase I and II) which were purified to homogeneity. Their molar mass, estimated by SDS-PAGE, were 21 and 24 kDa, respectively. Both enzymes are glycoproteins with 50°C temperature optimum; optimum pH was 6.0-6.5 for xylanase I and 6.0 for xylanase II. At 50°C xylanase I exhibited higher thermostability than xylanase II. Hg2+, Cu 2+ and SDS were strong inhibitors, 1,4-dithiothreitol stimulated the reaction of both enzymes. Both xylanases are xylan-specific; kinetic parameters indicated higher efficiency in the hydrolysis of oat spelts xylan. In hydrolysis of this substrate, xylotriose, xylotetraose and larger xylooligosaccharides were released and hence the enzymes were classified as endoxylanases.

Cytotoxicity of denture base resins: Effect of water bath and microwave postpolymerization heat treatments

Purpose: This study compared the effect of two postpolymerization heat treatments on the cytotoxicity of three denture base resins on L929 cells using 3H-thymidine incorporation and MTT assays. Materials and Methods: Sample disks of Lucitone 550, QC 20, and Acron MC resins were fabricated under aseptic conditions and stored in distilled water at 37°C for 48 hours. Specimens were then divided into three groups: (1) heat treated in microwave oven for 3 minutes at 500 W; (2) heat treated in water bath at 55°C for 60 minutes; and (3) no heat treatment. Eluates were prepared by placing three disks into a sterile glass vial with 9 mL of Eagle's medium and incubating at 37°C for 24 hours. The cytotoxic effect from the eluates was evaluated using the 3H-thymidine incorporation and MTT assays, which reflect DNA synthesis levels and cell metabolism, respectively. Results: The components leached from the resins were cytotoxic to L929 cells when 3H- thymidine incorporation assay was employed. In contrast, eluates from all resins revealed noncytotoxic effects as measured by MTT assay. For both MTT assay and 3H-thymidine incorporation, the heat treatments did not decrease the cytotoxicity of the materials tested. Conclusion: Resins were graded by 3H-thymidine incorporation assay as slightly cytotoxic and by MTT assay as noncytotoxic. Cytotoxicity of the denture base materials was not influenced by microwave or water bath heat treatment.

Avaliação da formação do biofilme de Candida albicans sobre superfície de titânio revestida com hidroxiapatita por meio de microscopia eletrônica de varredura

In this study Candida albicans biofilm formation on the surface of commercially pure titanium (cp-Ti) coated with hydroxyapatite was observed by means of scanning electron microscope. The biofilm was formed after 45 days of incubation of the samples in liquid culture medium inoculated with fungus cells in a tube of polystyrene with screw cap and sterilized. After the biofilm removal with 10% EDTA solution was observed pitting on the surface of cp-Ti coated.

Diffusion ability of endotoxin through dentinal tubules

The aim of this study was to evaluate the ability of endotoxin to diffuse through dentinal tubules towards the cement and to observe the period of time needed for it to reach the external root surface. Thirty single-rooted human teeth had their crowns and apices removed in order to standardize the root length to 15 mm. Teeth were instrumented until #30 K-file and made externally impermeable with epoxy adhesive, leaving 10 mm of the exposed root (middle third). The specimens were placed in plastic vials and irradiated (60Co gamma-rays). Then, they were divided into 2 groups (n = 15): G1) Escherichia coli endotoxin was inoculated into the root canal of the specimens and 1 ml of pyrogen-free water was put in the tubes; G2) (control): pyrogen-free water was inoculated into the root canals and 1 ml of pyrogen-free water was put in each tube. After 30 min, 2 h, 6 h, 12 h, 24 h, 48 h, 72 h and 7 days, the water of the tubes was removed and replaced. The removed aliquot was tested for the presence of endotoxin. Considering that the endotoxin is a B-lymphocyte polyclonal activator, at each experimental period, B-lymphocyte culture was stimulated with a sample of water removed from each tube and antibody (IgM) production was detected by ELISA technique. The results of IgM production were higher in groups of 24 h, 48 h, 72 h and 7 days in relation to the other studied groups, with statistically significant differences (ANOVA and Tukey's test p < 0.05). Endotoxin was able to diffuse through the dentinal tubules towards the cement, reaching the external root surface after the period of 24 h.

Synthesis, characterization, and biological activity of a new palladium(II) complex with deoxyalliin

Synthesis, characterization, and biological activity of a new water-soluble Pd(II)-deoxyalliin (S-allyl-L-cysteine) complex are described in this article. Elemental and thermal analysis for the complex are consistent with the formula [Pd(C6H10NO2S)2]. 13C NMR, 1H NMR, and IR spectroscopy show coordination of the ligand to Pd(II) through S and N atoms in a square planar geometry. Final residue of the thermal treatment was identified as a mixture of PdO and metallic Pd. Antiproliferative assays using aqueous solutions of the complex against HeLa and TM5 tumor cells showed a pronounced activity of the complex even at low concentrations. After incubation for 24 h, the complex induced cytotoxic effect over HeLa cells when used at concentrations higher than 0.40 mmol/L. At lower concentrations, the complex was nontoxic, indicating its action is probably due to cell cycle arrest, rather than cell death. In agreement with these results, the flow cytometric analysis indicated that after incubation for 24 h at low concentrations of the complex cells are arrested in G0/G1. © 2005 NRC Canada.

Inhibitory effect of deferoxamine on Paracoccidioides brasiliensis survival in human monocytes: Reversal by holotransferrin not by apotransferrin

The mechanisms used by Paracoccidioides brasiliensis to survive into phagocytic cells are not clear. Cellular iron metabolism is of critical importance to the growth of several intracellular pathogens whose capacity to multiply in mononuclear phagocytes is dependent on the availability of intracellular iron. Thus, the objective of this paper was to investigate the role of intracellular iron in regulating the capacity of P. brasiliensis yeast cells to survive within human monocytes. Treatment of monocytes with deferoxamine, an iron chelator, suppressed the survival of yeasts in a concentration-dependent manner. The effect of deferoxamine was reversed by iron-saturated transferrin (holotransferrin) but not by nonsaturated transferrin (apotransferrin). These results strongly suggest that P. brasiliensis survival in human monocytes is iron dependent.

Evaluation of serum- and animal protein-free media for the production of infectious bronchitis virus (M41) strain in a continuous cell line

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Micropropagação de Calendula officinalis L

Micropropagation of Calendula officinalis L. is usually propagated through seeds and therefore shows high diversity in flower size and colour, what causes quantitative and qualitative chemical variability. A micropropagation protocol was established for clonal propagation of this species to achieve homogeneous biomass, more appropriate for the production of phytotherapics. Explants harvested from capitula were the most appropriate for the micropropagation process. MS culture medium supplemented with 1.0 mgL-1 BAP and 6.0 gL-1 Phytagel™ enhanced shoot proliferation, while MS medium supplemented with 0.5 mgL-1 Kinetin and 6.0 gL-1 Phytagel™, increased shoot elongation. Plantlets (80%) cultured in MS/2 medium supplemented with 1.0 mgL-1 de IBA rooted.

Effectiveness of microwave irradiation on the disinfection of complete dentures

Purpose: The purpose of this study was to evaluate the effectiveness of microwave irradiation on the disinfection of simulated complete dentures. Materials and Methods: Eighty dentures were fabricated in a standardized procedure and subjected to ethylene oxide sterilization. The dentures were individually inoculated (10 7 cfu/mL) with tryptic soy broth (TSB) media containing one of the tested microorganisms (Candida albicans, Streptoccus aureus, Bacillus subtilis, and Pseudomonas aeruginosa). After 48 hours of incubation at 37°C, 40 dentures were individually immersed in 200 mL of water and submitted to microwave irradiation at 650 W for 6 minutes. Forty nonirradiated dentures were used as positive controls. Replicate aliquots (25 μL) of suspensions were plated at dilutions of 10 -3 to 10 -6 on plates of selective media appropriate for each organism. All plates were incubated at 37°C for 48 hours. TSB beakers with the microwaved dentures were incubated at 37°C for 7 more days. After incubation, the number of colony-forming units was counted and the data were statistically analyzed by Kruskal-Wallis test (α = .05). Results: No evidence of growth was observed at 48 hours for S aureus, B subtilis, and C albicans. Dentures contaminated with P aeruginosa showed small growth on 2 plates. After 7 days incubation at 37°C, no growth was visible in the TSB beakers of S aureus and C albicans. Turbidity was observed in 3 broth beakers, 2 from P aeruginosa and 1 from B subtilis. Conclusion: Microwave irradiation for 6 minutes at 650 W produced sterilization of complete dentures contaminated with S aureus and C albicans and disinfection of those contaminated with P aeruginosa and B subtilis.

Efeito da sacarose e sorbitol na conservação in vitro de Passiflora giberti N. E. Brown

This work objectified the study of sucrose and sorbitol effect in the in vitro conservation for Passiflora giberti N. E. Brown, access. Therefore, an experiment was conducted in a completely randomized design to compare control treatment (standard MS) to MS medium supplemented with three sucrose concentrations (0, 15 and 30 g L -1) combined with three sorbitol concentrations (10, 20 and 40 g L -1), in a total of 10 treatments with 20 replicas. The experiment evaluation was carried out at 30, 60, 90 and 120 days of incubation, whereas the height of shoots (cm), number of roots, number and color of leaves were observed. The results showed the possibility to maintain passion-fruit microplants for a four months period under slow growth in MS medium supplemented with 10 or 20 g L -1 of sorbitol, without sucrose, and kept under 16 hours photoperiod (22 μ E m -2 s -1) and temperature of 27 ± 1°C. Sucrose sustained the longest development of the microplants. Root formation was affected by the sorbitol in the concentration of 40 g L -1 and by the absence of sucrose in the culture medium.

Aspectos toxicológicos do bissulfito e outros agentes tóxicos sobre a síntese de glicerol-3-fosfato desidrogenase citoplasmá tica de levedura de panificação

The synthesis of intracellular glycerol-3-phosphate dehydrogenase (EC 1.1.1.8) in baker's yeast was investigated in submerged culture supplied with glucose or glycerol as sole carbon sources. Inhibitors of the glycolytic pathway, Krebs cycle and respiratory chain did not stimulate glycerol-3-phosphate dehydrogenase synthesis when added in low concentrations in up 7.5 × 10 -5 mol/L. The repression exercised by glucose on the synthesis of glycerol-3-phosphate dehydrogenase in YP-glucose medium was reduced by the addition of fermentation products and of sodium bisulfite. Synthesis of the enzyme was raised 22-110%. However, in YP-glycerol medium, the addition of 0.06% (w/v) sodium bisulfite reduced (29%) the synthesis of the enzyme, while 0.012% (v/v) acetaldehyde stimulated the synthesis of glycerol-3-phosphate dehydrogenase by 12%.

A specific short dextrin-hydrolyzing extracellular glucosidase from the thermophilic fungus Thermoascus aurantiacus 179-5

The thermophilic fungus Thermoascus aurantiacus 179-5 produced large quantities of a glucosidase which preferentially hydrolyzed maltose over starch. Enzyme production was high in submerged fermentation, with a maximal activity of 30 U/ml after 336 h of fermentation. In solid-state fermentation, the activity of the enzyme was 22 U/ml at 144 h in medium containing wheat bran and 5.8 U/ml at 48 h when cassava pulp was used as the culture medium. The enzyme was specific for maltose, very slowly hydrolyzed starch, dextrins (2-7G) and the synthetic substrate (α-PNPG), and did not hydrolyze sucrose. These properties suggest that the enzyme is a type II α-glucosidase. The optimum temperature of the enzyme was 70°C. In addition, the enzyme was highly thermostable (100% stability for 10 h at 60°C and a half-life of 15 min at 80°C), and stable within a wide pH range. Copyright © 2006, The Microbiological Society of Korea.

Espermidina e espermina exogena na micropropagação de Aloe vera (L.) Burm

The lateral shoots of the Aloe vera L. Burm have been inoculated in culture medium. The work has been divided in two phases. In the phase I different concentrations of NAA and BAP were added to the MS . In the phase II, plants proceeding of the MS were inoculated in medium containing or not spermidine and/or spermine. In the phase I the best results for mass production and shooting came out with the use of MS + 8,88 m mol L -1 BAP + 5,36 m mol L -1 NAA and these treatments didn't induce the rooting. Plants submitted to treatments with polyamines presented emission of roots during the phase II, suggesting possible inductor effect when used by isolated means during the rooting. No traces of oxidation were noted in the culture medium containing the polyamines. The use of spermidine in the culture-medium promoted a greater mass increasing and the most number of shoots was obtained with the use of the combination of polyamines. © 2008 1999, Editorial Ciencias Médicas.