Phosphorothioate antisense oligonucleotides induce the formation of nuclear bodies

Antisense oligonucleotides are powerful tools for the in vivo regulation of gene expression. We have characterized the intracellular distribution of fluorescently tagged phosphorothioate oligodeoxynucleotides (PS-ONs) at high resolution under conditions in which PS-ONs have the potential to display antisense activity. Under these conditions PS-ONs predominantly localized to the cell nucleus where they accumulated in 20-30 bright spherical foci designated phosphorothioate bodies (PS bodies), which were set against a diffuse nucleoplasmic population excluding nucleoli. PS bodies are nuclear structures that formed in cells after PS-ON delivery by transfection agents or microinjection but were observed irrespectively of antisense activity or sequence. Ultrastructurally, PS bodies corresponded to electron-dense structures of 150-300 nm diameter and resembled nuclear bodies that were found with lower frequency in cells lacking PS-ONs. The environment of a living cell was required for the de novo formation of PS bodies, which occurred within minutes after the introduction of PS-ONs. PS bodies were stable entities that underwent noticeable reorganization only during mitosis. Upon exit from mitosis, PS bodies were assembled de novo from diffuse PS-ON pools in the daughter nuclei. In situ fractionation demonstrated an association of PS-ONs with the nuclear matrix. Taken together, our data provide evidence for the formation of a nuclear body in cells after introduction of phosphorothioate oligodeoxynucleotides

Seismic evaluation of commercial plutonium fabrication plants in the United States

This report is an overview of Lawrence Livermore National Laboratory's seismic assessment of six commercial plutonium fabrication plants licensed by the US Nuclear Regulatory Commission (NRC) before September 2, 1971. The seismic assessment generally has three parts: (1) documentation of the structural condition of each facility and its critical equipment; (2) characterization of the seismic hazard (i.e., determination of peak ground acceleration vs return period for each site); and (3) evaluation of seismic capacity to determine ground motion levels at which critical structures and equipment fail. The failure evaluation used structural capacities of median-centered strength characteristics of the as-built configurations from (1) and seismic hazard input from (2). Results of the assessment were partial input for an overall natural risks study by the NRC

The Relationship of the Erythrocyte Sedimentation Rate to Inflammatory Cytokines and Survival in Patients With Chronic Heart Failure Treated With Angiotensin-Converting Enzyme Inhibitors

Objectives. The object of the study was to assess the relationship between erythrocyte sedimentation rate (ESR) and inflammatory cytokine production in chronic heart failure (CHF). Our findings lead us to re-evaluate the prognostic value of the ESR in assessing patients with CHF. Background. The search for simple prognostic markers in CHF that can be assessed anywhere at low cost is important. Increases in ESR are related to the acute phase response in states of inflammation and infection. Methods. Initially, we studied ESR in relation to plasma levels of inflammatory cytokines in 58 CHF patients. The findings prompted us to analyze the mortality predictive power of ESR compared with established risk factors in these patients and (retrospectively) in a second group of 101 clinically stable CHF patients who had ESR measured. Results. In all 159 CHF patients (age 62 ± 2 years, New York Heart Association [NYHA] class 2.7 ± 0.1), ESR ranged from 1 to 96 mm/h (median 14 mm/h). The ESR was correlated with tumor necrosis factor (TNF)-alpha (r = 0.31, p < 0.05), soluble TNF receptor-1 (r = 0.48, p < 0.0005), soluble TNF receptor-2 (r = 0.39, p < 0.005) and interleukin 6 (r = 0.45, p < 0.005) levels. High ESR levels indicated a poor prognosis (p < 0.0001), and this was independent of age, NYHA class, ejection fraction and peak oxygen consumption (p < 0.005). Patients with ESR above median (≥15 mm/h) compared with patients with ESR <15 mm/h had an impaired survival (hazard ratio 2.62, 95% confidence interval 1.58-4.36, p < 0.0001). Conclusions. Our study demonstrates that in CHF a high ESR is an unfavorable prognostic sign, independent of patients' symptomatology and ventricular function. These results are in diametrical contrast to previous results. This may reflect a change in the underlying pathophysiology due to today's treatment with angiotensin-converting enzyme inhibitors

Seasonal variations of dissolved nitrogen and DOC:DON ratios in an intermittent Mediterranean stream

22 páginas, 6 figuras, 4 tablas.Seasonal variations of dissolved inorganic nitrogen (DIN) (NO3–N and NH4–N) and dissolved organic nitrogen (DON) were determined in Fuirosos, an intermittent stream draining an unpolluted Mediterranean forested catchment (10.5 km2) in Catalonia (Spain). The influence of flow on streamwater concentrations and seasonal differences in quality and origin of dissolved organic matter, inferred from dissolved organic carbon to nitrogen ratios (DOC:DON ratios), were examined. During baseflow conditions, nitrate and ammonium had opposite behaviour, probably controlled by biological processes such as vegetation uptake and mineralization activity. DON concentrations did not have a seasonal trend. During storms, nitrate and DON increased by several times but discharge was not a good predictor of nutrient concentrations. DOC:DON ratios in streamwater were around 26, except during the months following drought when DOC:DON ratios ranged between 42 and 20 during baseflow and stormflow conditions, respectively. Annual N export during 2000–2001 was 70 kg km 1 year 1, of which 75% was delivered during stormflow. The relative contribution of nitrogen forms to the total annual export was 57, 35 and 8% as NO3–N, DON and NH4–N, respectively.This study was supported by a Formacio´n de Personal Investigador (FPI) grant and funds provided by the Comisio´ n Interministerial de Ciencia y Tecnologı´a (CICT, reference REN2001-3327).Peer reviewe