Epifluorescence and light microscopy evidencing structural and functional polymorphism of ribosomal DNA in fish (Teleostei: Astyanax fasciatus)

New karyotypic data are presented for the Astyanax fasciatus species complex from four different locations on the Upper Tibagi River in the state of Paraná, Brazil. Chromosome markers were analyzed using conventional (Ag-NOR) and molecular (FISH with 18S biotinylated probes) methods. Two cytotypes were found in cell counts with diploid number 2n = 48 chromosomes and 2n = 50 chromosomes, previously denominated Cytotype A and B, respectively. Two specific patterns of Ag-NORs markers (ribosomal gene activity) were found, with intra-population and inter-population variations. Cytotype A exhibited two to three chromosomes with NOR sites in the metaphases analyzed. In Cytotype B specimens, up to three markers were found, demonstrating greater intra-population and inter-population variation. All individuals with only one chromosome pair with NORs were located in the telomeric region of the short arm of Chromosome 5. This characteristic was interpreted as ancestral for the species. Another identified pattern revealed a site in the telomeric region probably in the long arm of Chromosome 4 and another submetacentric chromosome with bitelomeric marks exclusively in specimens with 2n = 50 chromosomes. In the FISH analysis (ribosomal gene structure), five to seven markers were identified in Cytotype A and three to seven markers were identified in Cytotype B. Structural chromosome events and/or transposable elements are required to explain the ribosomal gene location diversity in these organisms. The results of the present study corroborate the hypothesis that the A. fasciatus of the Upper Tibagi River region constitute a species complex.     

Direct determination of Cr and Cu in urine samples by electrothermal atomic absorption spectrometry using ruthenium as permanent modifier (R1)

In this study Ru, deposited thermally on an integrated platform pyrolytic graphite tube, is proposed as a permanent modifier for the determination of Cu and Cr in urine samples by electrothermal atomic absorption spectrometry. The samples were diluted 1:1 with nitric acid (1% v/v). Pyrolysis and atomization temperatures for spiked urine samples were 1,100 degrees C and 1,900 degrees C respectively for Cu, and 1,400 degrees C and 2,500 degrees C respectively for Cr. For comparison purposes, the conventional modification with Pd+Mg was also studied. The sensitivity for Ru as permanent modifier was higher for the two analytes. The characteristic masses were 7.3 and 17.7 for Cr and Cu. The detection limits (3sigma) were 0.22 and 0.32 microg/L, for Cr and Cu, respectively. Good agreement was obtained with certified urine samples for the two elements.     

Molecular imprinting of β-cyclodextrin/cholesterol template into a silica polymer for cholesterol separation

The molecular assembly formed by the inclusion complex of cholesterol in β-cyclodextrin (β-CD:chol) was used as a template for the molecular imprinting of a sol–gel polymer (MIP/β-CD:chol), produced with tetraethoxysilane (TEOS) as precursor. The MIP/β-CD:chol and pure silica matrix (PSM) were tested for the efficiency of cholesterol removal from solutions at different cholesterol concentrations (1–10 mg/mL). The adsorption tests were run at 25°C using 1% (w/v) solid/liquid suspensions during 24 h. The MIP/β-CD:chol data on cholesterol adsorption was fitted by the Langmüir isotherm model, giving a maximum adsorption capacity of 76.5 mg cholesterol/g-adsorbent. The PSM data did conform to the Langmüir model. The maximum cholesterol adsorption achieved with the PSM was higher, 251 mg/g, probably due to multilayer adsorption. The hydrophobic silica matrix, imprinted with the inclusion complex of β-CD and a target molecule, has the potential of being used as an adsorbent for other organic molecules.     

Rapid and sensitive method for the determination of acetaldehyde in fuel ethanol by high-performance liquid chromatography with UV–Vis detection

A high-performance liquid chromatography (HPLC) method for the determination of acetaldehyde in fuel ethanol was developed. Acetaldehyde was derivatized with 0.900 mL 2,4-dinitrophenylhydrazine (DNPHi) reagent and 50 L phosphoric acid 1 mol L^–1 at a controlled room temperature of 15°C for 20 min. The separation of acetaldehyde-DNPH (ADNPH) was carried out on a Shimadzu Shim-pack C₁₈ column, using methanol/LiCl_(aq) 1.0 mM (80/20, v/v) as a mobile phase under isocratic elution and UV–Vis detection at 365 nm. The standard curve of ADNPH was linear in the range 3–300 mg L^–1 per injection (20 L) and the limit of detection (LOD) for acetaldehyde was 2.03 g L^–1, with a correlation coefficient greater than 0.999 and a precision (relative standard deviation, RSD) of 5.6% (n=5). Recovery studies were performed by fortifying fuel samples with acetaldehyde at various concentrations and the results were in the range 98.7–102%, with a coefficient of variation (CV) from 0.2% to 7.2%. Several fuel samples collected from various gas stations were analyzed and the method was successfully applied to the analysis of acetaldehyde in fuel ethanol samples.     

Microwave-assisted synthesis of borneol esters and their antimicrobial activity

Seventeen borneol esters (1–17) were synthesised by conventional and microwave-assisted methodology using DIC/DMAP, and seven are described for the first time (8, 9, 10, 12, 13, 16 and 17). The microwave-assisted methodology was carried out without use of solvents, displayed short reaction times, and showed equal or higher yields for all the long-chain esters and three aromatic compounds (11, 12 and 14) when compared to the conventional approach. All the borneol esters were evaluated against the bacteria Streptococcus sanguinis, Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa and the fungus Candida albicans. Compounds 12, 13 and 14 displayed promising antibacterial activity with a MIC equal to ampicilin (62.5 mg mL^?1) for some microorganisms. In fact, bornyl 3′,4′-dimethoxybenzoate (13) was active against all tested bacteria and fungus.     

Isolation and structure determination of bioactive isoflavones from callus culture of Dipteryx odorata

Crude extracts of callus and roots of Dipteryx odorata were analyzed by HPLC to detect and quantify isoflavone contents. Based on spectroscopic and X-ray crystallography data the structures of two isoflavones were elucidated as 7-hydroxy-4',6-dimethoxyisoflavone and 3',7-dihydroxy-4',6-dimethoxyisoflavone. The production of dry biomass of 7-hydroxy-4',6-dimethoxyisoflavone in cultured callus was 4.12 mg/g, approximately eleven fold higher than the amount accumulated in roots of D. odorata wild-growing plants. The 7-hydroxy-4',6-dimethoxyisoflavone was effective against glyceraldehyde-3-phosphate dehydrogenase from Trypanosoma cruzi. This is the first report on those bioactive isoflavones accumulated in callus of D. odorata.     

Aqueous solution behavior of thermosensitive (N-isopropylacrylamide-acrylic acid-ethyl methacrylate) terpolymers

A series of N-isopropylacrylamide (NIPAM)-acrylic acid–ethyl methacrylate terpolymers with varied monomer compositions was prepared by radical polymerization. The solution behavior of these polymers was studied in dilute aqueous solution using spectrophotometry, fluorescence spectroscopy and high-sensitivity differential scanning calorimetry. The results obtained revealed that the lower critical solution temperatures depend strongly on the copolymer composition, solution pH and ionic strength. At a high pH, the ionization of acrylic acid (AA) units leads to an increase in solution cloud points (T_c). Solutions of polymers containing 10% or less of AA display a constant T_c for pH above 5.5, with 15% there is a continuous increase in T_c with pH and, for higher AA contents, no clouding was observed within the studied temperature range. Fluorescence probe studies were conducted by following the I ₁/I ₃ ratio of pyrene vibronic bands and the emission of anilinonaphtalene sulfonic acid, sodium salt (ANS), both approaches revealing the existence of hydrophobic domains for polymers with higher ethyl methacrylate content at temperatures lower than T_c, suggesting some extent of aggregation and/or a coil-to-globule transition. Scanning calorimetry measurements showed an endothermic transition at temperatures agreeing with the previously detected cloud points. Moreover, the transition curves became broader and with a smaller transition enthalpy, as both the AA content and the solution pH were increased. These broader transitions were interpreted to be the result of a wider molecular distribution upon polymer ionization, hence, displaying varied solution properties. The decrease in transition enthalpy was rationalized as a consequence of reminiscent hydration of NIPAM units, even after phase separation, owing to the presence of electric charges along the polymer chain.     

Photobiomodulation Therapy on the Guided Bone Regeneration Process in Defects Filled by Biphasic Calcium Phosphate Associated with Fibrin Biopolymer

The aim is to evaluate the effects of photobiomodulation therapy (PBMT) on the guided bone regeneration process (GBR) in defects in the calvaria of rats filled with biphasic calcium phosphate associated with fibrin biopolymer. Thirty male Wistar rats were randomly separated: BMG (n = 10), defects filled with biomaterial and covered by membrane; BFMG (n = 10), biomaterial and fibrin biopolymer covered by membrane; and BFMLG (n = 10), biomaterial and fibrin biopolymer covered by membrane and biostimulated with PBMT. The animals were euthanized at 14 and 42 days postoperatively. Microtomographically, in 42 days, there was more evident bone growth in the BFMLG, limited to the margins of the defect with permanence of the particles. Histomorphologically, an inflammatory infiltrate was observed, which regressed with the formation of mineralized bone tissue. In the quantification of bone tissue, all groups had a progressive increase in new bone tissue with a significant difference in which the BFMLG showed greater bone formation in both periods (10.12 ± 0.67 and 13.85 ± 0.54), followed by BFMG (7.35 ± 0.66 and 9.41 ± 0.84) and BMG (4.51 ± 0.44 and 7.11 ± 0.44). Picrosirius-red staining showed greater birefringence of collagen fibers in yellow-green color in the BFMLG, showing more advanced bone maturation. PBMT showed positive effects capable of improving and accelerating the guided bone regeneration process when associated with biphasic calcium phosphate and fibrin biopolymer.     

A new high-performance liquid chromatography assay for the determination of sesquiterpene lactone 15-deoxygoyazensolide in rat plasma

A simple, rapid and sensitive high-performance liquid chromatography method was developed for the analysis of the sesquiterpene lactone 15-deoxygoyazensolide (LAC15-D) in rat plasma samples. The chromatographic separation was achieved on a LiChrospher^® RP18 column using methanol:water (50:50, v/v) containing 0.6% acetic acid as mobile phase, at a flow rate of 0.7 mL min⁻¹. UV detection was carried out at 270 nm. Phenytoin was used as internal standard. Prior to the analysis, the rat plasma samples were submitted to liquid-liquid extraction with dichloromethane. The mean absolute recoveries were 73% with R.S.D. values lower than 3.5. The method was linear over the 6.0-2000 ng mL⁻¹ concentration range and the quantification limit was 6.0 ng mL⁻¹. Within-day and between-day assay precision and accuracy were studied at three concentration levels (15, 300 and 480 ng mL⁻¹) and were lower than 15%. The validated method was used to measure the plasmatic concentration of LAC15-D in rats that received a single intraperitoneal dose of 30 mg kg⁻¹.