Production of Bovine Hand-Made Cloned Embryos by Zygote-Oocyte Cytoplasmic Hemi-complementation

The aim of this study was to evaluate the effect of the cytoplast type and activation process on development of cloned embryos. Bovine oocytes (MII) or zygotes at the one-cell stage (IVF) were manually bisected and segregated in MII or IVF hemi-cytoplasts or hemi-karyoplasts. Adult skin cells from a bovine female were used as nucleus donors (SC). Experimental groups were composed of IVF embryos; parthenogenetic embryos; handmade cloned (HMC) embryos; and reconstructed HMC embryos using IVF hemi-cytoplast + MII hemi-cytoplast + SC (G-I); IVF hemi-cytoplast + IVF hemi-cytoplast + SC (G-II); MII hemi-cytoplast + IVF hemi-karyoplast (G-III); and IVF hemi-cytoplast + IVF hemi-karyoplast (G-IV). Embryos from G-I to G-IV were allocated to subgroups as sperm-activated (SA) or were further chemically activated (SA + CA). Embryos from all groups and subgroups were in vitro cultured in the WOW system. Blastocyst development in subgroup G-I SA (28.2%) was similar to IVF (27.0%) and HMC (31.4%) controls, perhaps due to a to a more suitable activation process and/or better complementation of cytoplasmic reprogramming factors, with the other groups and subgroups having lower levels of development. No blastocyst development was observed when using IVF hemi-karyoplasts (G-III and G-IV), possibly due to the manipulation process during a sensitive biological period. In summary, the presence of cytoplasmic factors from MII hemi-oocytes and the sperm activation process from hemi-zygotes appear to be necessary for adequate in vitro development, as only the zygote-oocyte hemi-complementation was as efficient as controls for the generation of bovine cloned blastocysts.

A novel locus for split-hand/foot malformation associated with tibial hemimelia (SHFLD syndrome) maps to chromosome region 17p13.1-17p13.3

Split-hand/foot malformation (SHFM) associated with aplasia of long bones, SHFLD syndrome or Tibial hemimelia-ectrodactyly syndrome is a rare condition with autosomal dominant inheritance, reduced penetrance and an incidence estimated to be about 1 in 1,000,000 liveborns. To date, three chromosomal regions have been reported as strong candidates for harboring SHFLD syndrome genes: 1q42.2-q43, 6q14.1 and 2q14.2. We characterized the phenotype of nine affected individuals from a large family with the aim of mapping the causative gene. Among the nine affected patients, four had only SHFM of the hands and no tibial defects, three had both defects and two had only unilateral tibial hemimelia. In keeping with previous publications of this and other families, there was clear evidence of both variable expression and incomplete penetrance, the latter bearing hallmarks of anticipation. Segregation analysis and multipoint Lod scores calculations (maximum Lod score of 5.03 using the LINKMAP software) using all potentially informative family members, both affected and unaffected, identified the chromosomal region 17p13.1-17p13.3 as the best and only candidate for harboring a novel mutated gene responsible for the syndrome in this family. The candidate gene CRK located within this region was sequenced but no pathogenic mutation was detected.

On the release of metronidazole from natural rubber latex membranes

The controlled release of drugs can be efficient if a suitable encapsulation procedure is developed, which requires biocompatible materials to hold and release the drug. In this study, a natural rubber latex (NRL) membrane is used to deliver metronidazole (MET), a powerful antiprotozoal agent. MET was found to be adsorbed on the NRL membrane, with little or no incorporation into the membrane bulk, according to energy dispersive X-ray spectroscopy. X-ray diffraction and FTIR spectroscopy data indicated that MET retained its structural and spectroscopic properties upon encapsulation in the NRL membrane, with no molecular-level interaction that could alter the antibacterial activity of MET. More importantly, the release time of MET in a NRL membrane in vitro was increased from the typical 6-8 h for oral tablets or injections to ca. 100 h. The kinetics of the drug release could be fitted with a double exponential function, with two characteristic times of 3.6 and 29.9 h. This is a demonstration that the induced angiogenesis known to be provided by NRL membranes can be combined with a controlled release of drugs, whose kinetics can be tailored by modifying experimental conditions of membrane fabrication for specific applications. (C) 2010 Elsevier B.V. All rights reserved.

Elastomeric Composites Based on Ethylene-Propylene-Diene Monomer Rubber and Conducting Polymer-Modified Carbon Black

Thermally stable elastomeric composites based on ethylene-propylene-diene monomer (EPDM) and conducting polymer-modified carbon black (CPMCB) additives were produced by casting and crosslinked by compression molding. CPMCB represent a novel thermally stable conductive compound made via ""in situ"" deposition of intrinsically conducting polymers (ICP) such as polyaniline or polypyrrole on carbon black particles. Thermogravimetric analysis showed that the composites are thermally stable with no appreciable degradation at ca. 300 degrees C. Incorporating CPMCB has been found to be advantageous to the processing of composites, as the presence of ICP lead to a better distribution of the filler within the rubber matrix, as confirmed by morphological analysis. These materials have a percolation threshold range of 5-10 phr depending on the formulation and electrical dc conductivity values in the range of 1 x 10(-3) to 1 x 10(-2) S cm(-1) above the percolation threshold. A less pronounced reinforcing effect was observed in composites produced with ICP-modified additives in relation to those produced only with carbon black. The results obtained in this study show the feasibility of this method for producing stable, electrically conducting composites with elastomeric characteristics. POLYM. COMPOS., 30:897-906, 2009. (C) 2008 Society of Plastics Engineers

Extraction of cellulose whiskers from cassava bagasse and their applications as reinforcing agent in natural rubber

In this work cassava bagasse, a by-product of cassava starch industrialization was investigated as a new raw material to extract cellulose whiskers. This by-product is basically constituted of cellulose fibers (17.5 wt%) and residual starch (82 wt%). Therefore, this residue contains both natural fibers and a considerable quantity of starch and this composition suggests the possibility of using cassava bagasse to prepare both starch nanocrystals and cellulose whiskers. In this way, the preparation of cellulose whiskers was investigated employing conditions of sulfuric acid hydrolysis treatment found in the literature. The ensuing materials were characterized by transmission electron microscopy (TEM) and X-ray diffraction experiments. The results showed that high aspect ratio cellulose whiskers were successfully obtained. The reinforcing capability of cellulose whiskers extracted from cassava bagasse was investigated using natural rubber as matrix. High mechanical properties were observed from dynamic mechanical analysis. (C) 2010 Elsevier B.V. All rights reserved.

Phenolic matrices and sisal fibers modified with hydroxy terminated polybutadiene rubber: Impact strength, water absorption, and morphological aspects of thermosets and composites

The aim of the present work was to investigate the toughening of phenolic thermoset and its composites reinforced with sisal fibers, using hydroxyl-terminated polybutadiene rubber (HTPB) as both impact modifier and coupling agent. Substantial increase in the impact strength of the thermoset was achieved by the addition 10% of HTPB. Scanning electron microscopy (SEM) images of the material with 15% HTPB content revealed the formation of some rubber aggregates that reduced the efficiency of the toughening mechanism. In composites, the toughening effect was observed only when 2.5% of HTPB was added. The rubber aggregates were found located mainly at the matrix-fiber interface suggesting that HTPB could be used as coupling agent between the sisal fibers and the phenolic matrix. A composite reinforced with sisal fibers pre-impregnated with HTPB was then prepared; its SEM images showed the formation of a thin coating of HTPB on the surface of the fibers. The ability of HTBP as coupling agent between sisal fibers and phenolic matrix was then investigated by preparing a composite reinforced with sisal fibers pre-treated with HTPB. As revealed by its SEM images, the HTPB pre-treatment of the fibers resulted on the formation of a thin coating of HTPB on the surface of the fibers, which provided better compatibility between the fibers and the matrix at their interface, resulting in a material with low water absorption capacity and no loss of impact strength. (C) 2009 Elsevier B.V. All rights reserved.

Graphite-silicone rubber composite electrode: Preparation and possibilities of analytical application

Composite electrodes were prepared using graphite powder and silicone rubber in different compositions. The use of such hydrophopic materials interned to diminish the swallowing observed in other cases when the electrodes are used in aqueous solutions for a long time. The composite was characterized for the response reproducibility, ohmic resistance, thermal behavior and active area. The voltammetric response in relation to analytes with known voltammetric behavior was also evaluated, always in comparison with the glassy carbon. The 70% (graphite, w/w) composite electrode was used in the quantitative determination of hydroquinone (HQ) in a DPV procedure in which a detection limit of 5.1 x 10(-8) mol L-1 was observed. HQ was determined in a photographic developer sample with errors lower then 1% in relation to the label value. (c) 2007 Elsevier B.V. All rights reserved.

THE POTENTIALITIES OF USING A GRAPHITE-SILICONE RUBBER COMPOSITE ELECTRODE IN THE DETERMINATION OF PROPRANOLOL

A graphite silicone-rubber composite electrode (GSR) was used for the determination of propranolol in drug formulation. Cyclic voltammetry (CV) at the GSR presented an irreversible oxidation peak at + 0.8V vs. SCE, in Britton Robinson (B-R) buffer pH 7.4. The quantitative determination was carried out using differential pulse voltammetry (DPV). Under optimized parameters a linear dynamic range from 5.0 to 80.6 mu mol L(-1) with a detection limit of 1.1 mu mol L(-1) was observed. A repeatability of 4.5 +/- 0.1 mu A (n = 10) peak current was found after 10 successive DPV voltammograms of propranolol in the same solution after surface renovations. Using the proposed electrode, propranolol was quantified in a pharmaceutical formulation with results that agreed within 95% confidence level (t-test) with those from an official method.

Analytical Potentialities of Carbon Nanotube/Silicone Rubber Composite Electrodes: Determination of Propranolol

A new composite electrode based on multiwall carbon nanotubes (MWCNT) and silicone-rubber (SR) was developed and applied to the determination of propranolol in pharmaceutical formulations. The effect of using MWCNT/graphite mixtures in different proportions was also investigated. Cyclic voltammetry and electrochemical impedance spectroscopy were used for electrochemical characterization of different electrode compositions. Propranolol was determined using MWCNT/SR 70% (m/m) electrodes with linear dynamic ranges up to 7.0 mu molL(-1) by differential pulse and up to 5.4 mu molL(-1) by square wave voltammetry, with LODs of 0.12 and 0.078 mu molL(-1), respectively. Analysis of commercial samples agreed with that obtained by the official spectrophotometric method. The electrode is mechanically robust and presented reproducible results and a long useful life.