Fine mapping and genomic structure of ACTN2, the human gene coding for the sarcomeric isoform of alpha-actinin-2, expressed in skeletal and cardiac muscle.

The present paper reports on the fine mapping of the ACTN2 gene and on the reconstruction of its genomic structure. By radiation hybrid mapping, the gene was located about 912 cR from the 1p-telomere. ACTN2 was placed between the marker WI-9317 (alias D1S2421) and the marker AFMA045ZC5, within the chromosomal band 1q43. The gene was detected in YAC 955 c 12. This YAC was used as template DNA for long-distance and Alu-PCR, using a set of putative exonic primers, designed on the cDNA sequence of alpha-actinin-2, in order to characterize the ACTN2 intron-exon boundaries. The entire genomic structure of the gene was reconstructed. The ACTN2 gene contained 21 exons, in a segment spanning about 40 kb of genomic DNA. Only the proximal part of the gene shows a high conservation through evolution, whereas in the remaining part a divergence from the genomic organization of C. elegans and D. melanogaster was noticed. A series of intronic primers was specifically designed and produced, to amplify all the exons of ACTN2, directly from genomic DNA. This will enable mutation screening in patients affected with hereditary diseases linked to the marker CA4F/R, a polymorphism in the last intron of the alpha-actinin-2 gene.     

Inhibition of oxidative metabolism of tocopherols with omega-N-heterocyclic derivatives of vitamin E

The oxidative metabolism of tocopherols and tocotrienols by monooxygenases is a key factor in the plasma and tissue clearance of forms of vitamin E other than alpha-tocopherol. It is well known that a commonly ingested form of vitamin E, gamma-tocopherol, has greatly reduced plasma half-life (faster clearance) than alpha-tocopherol. The tocotrienols are metabolized even faster than gamma-tocopherol. Both gamma-tocopherol and alpha- and delta-tocotrienol possess intriguing biological activities that are different from alpha-tocopherol, making them potentially of interest for therapeutic use. Unfortunately, the fast clearance of non-alpha-tocopherols from animal tissues is a significant hurdle to maximizing their effect(s) as dietary supplements. We report here the design and synthesis of N-heterocycle-containing analogues of alpha-tocopherol that act as inhibitors of Cyp4F2, the key monooxygenase responsible for omega-hydroxylation of the side chain of tocols. In particular, an omega-imidazole containing compound, 1, [(R)-2-(9-(1H-imidazol-1-yl)nonyl)-2,5,7,8-tetramethylchroman-6-ol] had an ED(50) for inhibition of gamma-CEHC production from gamma-tocopherol of approximately 1 nM when tested in HepG2 cells in culture. Furthermore, feeding of 1 to mice along with rapidly metabolized delta-tocopherol, resulted in a doubling of the delta-tocopherol/alpha-tocopherol ratio in liver (P<0.05). Thus, 1 may be a useful adjuvant to the therapeutic use of non-alpha-tocopherols.     

Amblyomma boeroi n. sp. (Acari: Ixodidae), a parasite of the Chacoan peccary Catagonus wagneri (Rusconi) (Artiodactyla: Tayassuidae) in Argentina

All parasitic stages of Amblyomma boeroi n. sp. (Acari: Ixodidae) are described here from Catagonus wagneri (Rusconi) in Argentina. The diagnostic characters for the male are a combination of orbited eyes, a 2/2 dental formula, coxa IV considerably larger than coxae I–III and with a long, sickle-shaped, medially directed spur arising from its internal margin, a scutum which is light grey to very pale ivory in colour, and the absence of a postanal groove. The diagnostic characters for the females are a combination of orbited eyes, a central pair and two marginal pairs of short, coarse notal setae, a 2/2 dental formula, and the absence of a postanal groove. The nymph has short palpi and a 2/2 dental formula arranged in 6 rows, its eyes are convex and orbited, and it has no postanal groove. The dorsally rectangular basis capituli of the larva, its bulging eyes and slightly sinuous posterior scutal margin all serve to distinguish it from the larva of other species of the genus. The principal host for all parasitic stages is C. wagneri (Artiodactyla: Tayassuidae). Phylogenetically A. boeroi appears to represent an independent lineage within Amblyomma Koch, 1844.     

Preparation of fluorescent tocopherols for use in protein binding and localization with the alpha-tocopherol transfer protein

Sixteen fluorescent analogues of the lipid-soluble antioxidant vitamin alpha-tocopherol were prepared incorporating fluorophores at the terminus of omega-functionalized 2-n-alkyl-substituted chromanols (1a-d and 4a-d) that match the methylation pattern of alpha-tocopherol, the most biologically active form of vitamin E. The fluorophores used include 9-anthroyloxy (AO), 7-nitrobenz-2-oxa-1,3-diazole (NBD), N-methyl anthranilamide (NMA), and dansyl (DAN). The compounds were designed to function as fluorescent reporter ligands for protein-binding and lipid transfer assays. The fluorophores were chosen to maximize the fluorescence changes observed upon moving from an aqueous environment (low fluorescence intensity) to an hydrophobic environment such as a protein's binding site (high fluorescence intensity). Compounds 9d (anthroyloxy) and 10d (nitrobenzoxadiazole), having a C9-carbon chain between the chromanol and the fluorophore, were shown to bind specifically and reversibly to recombinant human tocopherol transfer protein (alpha-TTP) with dissociation constants of approximately 280 and 60 nM, respectively, as compared to 25 nM for the natural ligand 2R,4'R,8'R-alpha-tocopherol. Thus, compounds have been prepared that allow the investigation of the rate of alpha-TTP-mediated inter-membrane transfer of alpha-tocopherol and to investigate the mechanism of alpha-TTP function at membranes of different composition.     

3D Stationary electric current density in a spherical tumor treated with low direct current: an analytical solution

Electrotherapy with direct current delivered through implanted electrodes is used for local control of solid tumors in both preclinical and clinical studies. The aim of this research is to develop a solution method for obtaining a three-dimensional analytical expression for potential and electric current density as functions of direct electric current intensity, differences in conductivities between the tumor and the surrounding healthy tissue, and length, number and polarity of electrodes. The influence of these parameters on electric current density in both media is analyzed. The results show that the electric current density in the tumor is higher than that in the surrounding healthy tissue for any value of these parameters. The conclusion is that the solution method presented in this study is of practical interest because it provides, in a few minutes, a convenient way to visualize in 3D the electric current densities generated by a radial electrode array by means of the adequate selection of direct current intensity, length, number, and polarity of electrodes, and the difference in conductivity between the solid tumor and its surrounding healthy tissue.     

Natural infestation of Hydrochoerus hydrochaeris by Amblyomma dubitatum ticks

Natural infestation of Amblyomma dubitatum in relation to individual specific attributes of Hydrochoerus hydrochaeris such as sex, body mass and body condition was analyzed. The anatomical distribution of A. dubitatum on H. hyrochaeris was also evaluated. Prevalence of adults and nymphs were significantly higher than prevalence of larvae. Non-significant differences in the infestation levels were found among host sex. Multiple regression analysis did not show any statistically significant association among the level of infestation with ticks and body mass and body condition of the host. All parasitic tick stages were collected in all five anatomical areas of the host, but they exhibited significant differences in feeding site preference. Factors associated to the host which determine the high levels of infestation with A. dubitatum could be assigned to a combination of population-level properties of the host as abundance, ubiquity and aggregation, rather than individual specific attributes related to body condition, body mass or sex.