Sequence Features and Transcriptional Stalling within Centromere DNA Promote Establishment of CENP-A Chromatin

Centromere sequences are not conserved between species, and there is compelling evidence for epigenetic regulation of centromere identity, with location being dictated by the presence of chromatin containing the histone H3 variant CENP-A. Paradoxically, in most organisms CENP-A chromatin generally occurs on particular sequences. To investigate the contribution of primary DNA sequence to establishment of CENP-A chromatin in vivo, we utilised the fission yeast Schizosaccharomyces pombe. CENP-ACnp1 chromatin is normally assembled on ∼10 kb of central domain DNA within these regional centromeres. We demonstrate that overproduction of S. pombe CENP-ACnp1 bypasses the usual requirement for adjacent heterochromatin in establishing CENP-ACnp1 chromatin, and show that central domain DNA is a preferred substrate for de novo establishment of CENP-ACnp1 chromatin. When multimerised, a 2 kb sub-region can establish CENP-ACnp1 chromatin and form functional centromeres. Randomization of the 2 kb sequence to generate a sequence that maintains AT content and predicted nucleosome positioning is unable to establish CENP-ACnp1 chromatin. These analyses indicate that central domain DNA from fission yeast centromeres contains specific information that promotes CENP-ACnp1 incorporation into chromatin. Numerous transcriptional start sites were detected on the forward and reverse strands within the functional 2 kb sub-region and active promoters were identified. RNAPII is enriched on central domain DNA in wild-type cells, but only low levels of transcripts are detected, consistent with RNAPII stalling during transcription of centromeric DNA. Cells lacking factors involved in restarting transcription-TFIIS and Ubp3-assemble CENP-ACnp1 on central domain DNA when CENP-ACnp1 is at wild-type levels, suggesting that persistent stalling of RNAPII on centromere DNA triggers chromatin remodelling events that deposit CENP-ACnp1. Thus, sequence-encoded features of centromeric DNA create an environment of pervasive low quality RNAPII transcription that is an important determinant of CENP-ACnp1 assembly. These observations emphasise roles for both genetic and epigenetic processes in centromere establishment

High eccentric strength training reduces heart rate variability in healthy older men

Background: Evaluation of non-pharmacological therapies that improve autonomic control of the heart rate in older subjects has a clinical significance, because reduced heart rate variability (HRV) can be associated with higher cardiovascular morbidity and mortality rates. Objective: To investigate if strength training improves cardiac autonomic control in healthy older men. Methods: The HRV of nine older healthy men (mean age 62 (2.0) years) was evaluated before and after 12 weeks of isokinetic eccentric strength training (2 days/week, 2-4 sets of 8-12 repetitions at 75-80% peak torque, involving knee flexion and extension. Electrocardiogram was continuously recorded for 15 min at rest, in supine and seated positions, before and after the strength training period. To estimate strength gains, the eccentric peak torque of the dominant leg was measured at 60 degrees/s by the same isokinetic dynamometer. Results: Mean systolic blood pressure decreased (123.78 (8.3) to 117.67 (10.2) mmHg, p < 0.05) and peak torque increased ( extension 210.02 (38.5) to 252.71 (60.9) N. m; flexion: 117.56 (25.1) to 132.96 (27.3) N. m, p < 0.05) after the strength training. The frequency domain indices showed a significant training effect (p < 0.05), since low frequency in normalised units and low frequency/ high frequency ratio increased (supine, 57 (14) to 68 (14), 1.56 (0.85) to 2.35 (1.48); seated, 65 (15) to 74 (8.0), 2.48 (1.09) to 3.19 (1.31), respectively), and high frequency in normalised units decreased (supine, 43 (14) to 32 (14); seated, 35 (15) to 26 (8)) after the training period. Conclusion: The results of the present investigation suggest that high eccentric strength training performed by healthy older men increases peak torque and reduces systolic blood pressure. However, an autonomic imbalance towards sympathetic modulation predominance was induced by an unknown mechanism.42

Economic viability of the piaucu Leporinus macrocephalus (Garavello & Britski, 1988) production

Brazilian fish farms presented an accelerated development during the early 90's, mainly because of the increase in fee-fishing operations. To meet the demand of this market, fish production and supply became excessive and, as a consequence, the number of fee-fishing operations, farmers and the final selling price, decreased. This study analyzes the technical aspects, production cost, profitability and economic viability of the production of piaucu (L. macrocephalus) in ponds, based on information from a rural property. Feeding and fingerling costs amount to approximately 47.1 % of the total production cost, representing together with the final selling price the most important factor affecting profitability. The payback period was 8.3 years, the liquid present value US$ 291.07, the internal return margin 9%, and the income-outcome ratio was 1.01, which represents an unattractive investment as a projection based on current conditions. The improvement in productive efficiency enhances the economic valuation index, and that the relative magnitude of cost and income are the most important points for the economic viability of the studied farm