Optimal tolerability of ultra-low-dose continuous combined 17beta-estradiol and norethisterone acetate: laboratory and safety results.

OBJECTIVE: To evaluate the influence of two ultra-low doses of oral continuous combined hormone therapy and placebo on metabolic parameters, and to assess safety endpoints and overall tolerability in healthy postmenopausal women. DESIGN: In a subpopulation of the Clinical study on Hormone dose Optimisation In Climacteric symptoms Evaluation (CHOICE) trial, lipids and parameters of glucose metabolism and hemostasis were analyzed in Nordic women (n = 158) at baseline and after 12 and 24 weeks of treatment with 0.5 mg 17beta-estradiol (E2) + 0.25 mg norethisterone acetate (NETA), 0.5 mg E2 + 0.1 mg NETA or placebo. Adverse events occurring from the first trial-related activity, whether related or not related to the study medication, were recorded for the entire population (n = 575) of the trial. The seriousness, relationship to treatment and the reason for withdrawal were reported. RESULTS: Both ultra-low-dose combinations were neutral to changes in lipid and lipoprotein, hemostasis parameters and carbohydrate metabolism during the trial. The incidence of serious adverse events was only 1% in respective treatment groups. Adverse events were the reason for withdrawal in only 2% and 6% of women in the 0.5 mg E2 + 0.25 mg and 0.1 mg NETA groups, and in 8% in the placebo group. No weight gain or change in blood pressure was reported during the trial in any of the study groups. CONCLUSION: The treatments had neutral effects on metabolic parameters in the study population. Excellent tolerability of both ultra-low doses resulted in high completion rates.

Modelling the effect of climate change on recovery of acidified freshwaters: relative sensitivity of individual processes in the MAGIC model.

The MAGIC model was used to evaluate the relative sensitivity of several possible climate-induced effects on the recovery of soil and surface water from acidification. A common protocol was used at 14 intensively studied sites in Europe and eastern North America. The results show that several of the factors are of only minor importance (increase in pCO(2) in soil air and runoff, for example), several are important at only a few sites (seasalts at near-coastal sites, for example) and several are important at nearly all sites (increased concentrations of organic acids in soil solution and runoff, for example). In addition changes in forest growth and decomposition of soil organic matter are important at forested sites and sites at risk of nitrogen saturation. The trials suggest that in future modelling of recovery from acidification should take into account possible concurrent climate changes and focus specially on the climate-induced changes in organic acids and nitrogen retention.

Biochemical properties of a hamster syncytium-forming ("foamy") virus.

A hamster syncytium-forming ("foamy") virus (HFV) was characterized. The HFV sedimented in isopyknic sucrose density gradients at 1.16-1.165 g/ml. It had RNA but no DNA, its replication was inhibited by actinomycin D, and it contained virion-associated, RNA-dependent DNA polymerase. Analysis of the RNA from purified virus showed several species: 62S, 40S, 28-30S, 18-20S, and 4-7S.

Modelling inorganic nitrogen in runoff: Seasonal dynamics at four European catchments as simulated by the MAGIC model.

Nitrogen (N) deposition is globally considered as a major threat to ecosystem functioning with important consequences for biodiversity, carbon sequestration and N retention. Lowered N retention as manifested by elevated concentrations of inorganic N in surface waters indicates ecosystem N saturation. Nitrate (NO3) concentrations in runoff from semi-natural catchments typically show an annual cycle, with low concentrations during the summer and high concentrations during the winter. Process-oriented catchment-scale biogeochemical models provide tools for simulation and testing changes in surface water and soil chemistry in response to changes in sulphur (S) and N deposition and climate. Here we examine the ability of MAGIC to simulate the observed monthly as well as the long-term trends over 10-35 years of inorganic N concentrations in streamwaters from four monitored headwater catchments in Europe: Certovo Lake in the Czech Republic, Afon Gwy at Plynlimon, UK, Storgama, Norway and G2 NITREX at Gårdsjön, Sweden. The balance between N inputs (mineralization+deposition) and microbial immobilization and plant uptake defined the seasonal pattern of NO3 leaching. N mineralization and N uptake were assumed to be governed by temperature, described by Q10 functions. Seasonality in NO3 concentration and fluxes were satisfactorily reproduced at three sites (R2 of predicted vs. modelled concentrations varied between 0.32 and 0.47 and for fluxes between 0.36 and 0.88). The model was less successful in reproducing the observed NO3 concentrations and fluxes at the experimental N addition site G2 NITREX (R2=0.01 and R2=0.19, respectively). In contrast to the three monitored sites, Gårdsjön is in a state of change from a N-limited to N-rich ecosystem due to 20 years of experimental N addition. At Gårdsjön the measured NO3 seasonal pattern did not follow typical annual cycle for reasons which are not well understood, and thus not simulated by the model. CAPSULE: The MAGIC model is able to simulate NO3 leaching on a monthly as well as an annual basis, and thus to reproduce the seasonal and short-term variations in N dynamics.

Modeling and monitoring of long-term acidification in an upland catchment of the Western Ore Mountains, SE Germany.

Modeling and monitoring of acidification patterns in a limed forest catchment in the Ore Mountains, SE Germany are presented. A lumped-parameter model, MAGIC, satisfactorily reproduced the main parameters of stream water chemistry. Despite repeated whole-catchment liming, between 1993 and 1999 stream water pH increased from 4.3 to only 4.6, while calcium concentrations declined slightly. Stream water sulfate concentration declined from 687 to 396 microeq l(-1), and the pronounced effect of deposition decline during the 1990s was probably more important than liming.

Buffering processes in a boreal dissolved organic carbon-rich stream during experimental acidification.

The role of organic acids on surface water acidity as well as their buffering during anthropogenic acidification and subsequent recovery was studied in a field experiment on a total organic carbon (TOC)-rich stream draining the Svartberget catchment in northern Sweden. H(2)SO(4) was added to the stream to increase SO(4)(2-) concentration by 90 microeq l(-1) for 30 h. About 60% of the added H(+) was buffered by protonation of organic acids, another 20% was buffered by base cations released from the surface of the stream channel and only ca. 20% of the added acid remained unbuffered. TOC concentrations (27 mg l(-1)), and site density of carboxylic groups--8.6 microeq (mg TOC)(-1)--remained stable during the experiment. Two models of organic acid dissociation (a triprotic model and a monoprotic pH-dependent pKa model) were fitted to the experimental results. These models explained the observed variations in organic anions, but the model parameters were quite different from those reported by studies in Northern America and Central Europe. This experiment had substantially more buffering effect of TOC between pH 4.4 and 5.3, which is an environmentally important pH range.

[Sentinel node biopsy in our department]. / Biopsie sentinelové uzliny na nasem pracovisti.

The author presents a group of two female patients with breast cancer and a male patient with malignant melanoma where mainly due to a borrowed scintillation tube combined with preoperative administration of methylene blue a sentinel node was successfully extirpated. The author analyzes the technical aspect of the biopsy. He is convinced that by combining the two methods its is within the powers of a district department to implement the method successfully.

Modelling soil nitrogen: the MAGIC model with nitrogen retention linked to carbon turnover using decomposer dynamics.

We present a new formulation of the acidification model MAGIC that uses decomposer dynamics to link nitrogen (N) cycling to carbon (C) turnover in soils. The new model is evaluated by application to 15-30 years of water chemistry data at three coniferous-forested sites in the Czech Republic where deposition of sulphur (S) and N have decreased by >80% and 40%, respectively. Sulphate concentrations in waters have declined commensurately with S deposition, but nitrate concentrations have shown much larger decreases relative to N deposition. This behaviour is inconsistent with most conceptual models of N saturation, and with earlier versions of MAGIC which assume N retention to be a first-order function of N deposition and/or controlled by the soil C/N ratio. In comparison with earlier versions, the new formulation more correctly simulates observed short-term changes in nitrate leaching, as well as long-term retention of N in soils. The model suggests that, despite recent deposition reductions and recovery, progressive N saturation will lead to increased future nitrate leaching, ecosystem eutrophication and re-acidification.

Anthropogenic acidification effects in primeval forests in the Transcarpathian Mts., western Ukraine.

The precipitation chemistry, deposition, nutrient pools and composition of soils and soil water, as well as an estimate of historical deposition of sulphur (S) and inorganic nitrogen (N) for the period 1860-2008, were determined in primeval deciduous and coniferous forests at the sites Javornik and Pop Ivan, respectively. Measured S throughfall inputs of 10 kg ha(-1)year(-1) in 2008 were similar to those estimated for the period 1900-1950 at both sites. The highest estimated S inputs were in the 1980s. Measured bulk deposition of N in 2008 was lower at Pop Ivan (5.6 kg ha(-1)year(-1)) compared to Javornik (12 kg ha(-1)year(-1)). Significantly lower NO(3) deposition was both estimated and measured at Pop Ivan. Higher soil base cation concentrations were observed at well-buffered Javornik underlain by flysch (Ca pool of 2046 kg ha(-1) and base saturation of 29%) compared to Pop Ivan underlain by crystalline schist (Ca pool of 186 kg ha(-1) and base saturation of 6.5%). The soil pool of organic carbon (C) was higher at Pop Ivan (212 t ha(-1)) compared to Javornik (127 t ha(-1)). The C concentration was positively correlated with organic N in the soil (p<0.001) at both sites, but the mass average C/N ratio in the forest floor was lower at Javornik (22) than at Pop Ivan (26). High N leaching of 17 kg ha(-1)year(-1) at the 90 cm depth was measured in the soil water at Javornik, suggesting high mineralization and nitrification rates in old growth deciduous forests in the area. Despite relatively low Al concentrations in the soil water, a low soil water Bc/Al ratio (0.9) (Bc=Ca+Mg+K) was found in the upper mineral soil at Pop Ivan. This suggests that the spruce forest ecosystems in the area are vulnerable to anthropogenic acidification and to the adverse effects of Al on forest root systems.

Retrospective analyses and future predictions of snowmelt-induced acidification: example from a heavily impacted stream in the Czech Republic.

We have combined a long-term hydrochemistry model (MAGIC) with a model that predicts short-term transient changes in hydrochemistry (pBDM) during hydrological events in order to improve the temporal resolution of retrospective analyses and future predictions of streamwater acidification. The model has been applied to a heavily impacted catchment in the Czech Republic. Spring flood acid-neutralizing capacity (ANC), pH, and inorganic monomeric aluminum (Ali(n+)) were simulated for the years of 1860, 1900, 1930, 1950, 1965, and 1985, measured in 1999, and predicted for 2030 using two different emission control scenarios. If the emission reduction according to the current legislation scenario is implemented, the model predicts that the spring flood pH, ANC, and Ali(n+) will recover close to the level of the 1950s by 2030. This will occur despite the annual average chemistry being farfrom having recovered to that level. The results suggest that the recovery of spring flood events is faster then the recovery of annual average chemistry and that much of what is won by further emission reduction will not be fully realized on an annual time scale.

Aberrant forms of Escherichia coli in blood cultures: in vitro reproduction of an in vivo observation.

Aberrant filamentous forms of Escherichia coli were observed on direct Gram stain of blood cultures from a patient being treated with the beta-lactam antibiotic cephalexin. After the institution of an alternative antibiotic regimen which included a different cell wall-active agent, E. coli of normal morphology was detected in blood cultures for an additional 48 h. Filamentous forms of E. coli could be reproduced reliably in vitro by incubating the organism in Mueller-Hinton broth containing various concentrations of cephalexin. Both supra- and subinhibitory concentrations of cephalexin resulted in filament formation after 4 h of incubation, whereas 24 h of incubation yielded intact filaments at only a narrow range of subinhibitory concentrations of cephalexin. In vitro comparison of the ability of cephalexin, cephalothin, ampicillin, and gentamicin to cause filamentous forms of E. coli showed that cephalexin and cephalothin produced pure filament formation after 4 h of incubation at subinhibitory concentrations of as low as one-fourth the minimum inhibitory concentration of the antibiotic. Ampicillin was not associated with pure filament formation at concentrations below the minimum inhibitory concentration, and gentamicin produced no filaments at any concentration. The effect of preincubation of E. coli with subinhibitory concentrations of cephalexin on subsequent minimum inhibitory concentrations of ampicillin was examined in an effort to develop an explanation for the persistent sepsis exhibited by the patient. No diminution of the activity of ampicillin by preincubation with cephalexin could be demonstrated. Other possible clinical implications of filamentous forms of gram-negative bacilli are discussed.

Characterization of RNA polymerase products of Nebraska calf diarrhea virus and SA11 rotavirus.

The endogenous RNA polymerase of the calf rotavirus was shown to synthesize single-stranded RNA transcripts of one polarity which were identical in size to the denatured parental double-stranded RNA segments. The transcripts were notable in their absence of polyadenylate sequences. The polymerase activity associated with the cores of calf rotavirus was minimally altered by the reaction, since calf rotavirus cores could be reused after purification, whereas the RNA polymerase products appeared to be released from the virion.

Monoclonal antibodies to respiratory syncytial virus proteins: identification of the fusion protein.

Six monoclonal antibodies directed against respiratory syncytial virus proteins were produced. Each was characterized by immunoprecipitation and indirect immunofluorescence. One was directed against the nucleocapsid protein. NP 44, two were directed against a 37,000-dalton protein, two were directed against the major envelope glycoprotein, GP 90, and one was directed against the 70,000-dalton envelope protein, VP 70. Indirect immunofluorescence stain patterns of infected HEp-2 cells defined GP 90 and VP 70 as viral proteins expressed on the cell surface, whereas NP 44 and the 37,000-dalton protein were detected as intracytoplasmic inclusions. One of the anti-GP 90 antibodies neutralized virus only in the presence of complement but did not inhibit cell-cell fusion. The anti-VP 70 antibody neutralized virus without complement and inhibited cell-cell fusion of previously infected HEp-2 cells, thus identifying VP 70 as the fusion protein.

Respiratory syncytial virus proteins: identification by immunoprecipitation.

The proteins of respiratory syncytial virus have not been clearly identified due to the lability of the virus and difficulties in its purification. We have pulse-labeled respiratory syncytial virus with [35S]methionine and [35S]cysteine and analyzed cell lysates by polyacrylamide gel electrophoresis. Five 35S-labeled viral proteins ranging in molecular weight from 21,000 to 73,000 (VP73, VP44, VP35, VP28, and VP21) were easily discernable above background cellular proteins. Treatment of the infected cells with 0.15 M NaCl before labeling suppressed host cell protein synthesis and allowed clearer visualization of the five viral proteins by polyacrylamide gel electrophoresis. Three glycoproteins (VGP 92, VGP 50, and VGP 17) were also identified after labeling with [3H]glucosamine. Five of these polypeptides (VP51, VP44, VP35, VP28, and VGP92) were shown to be antigenically active because they could be immunoprecipitated with anti-respiratory syncytial virus antibody produced in New Zealand white rabbits, cotton rats, and humans before analysis by polyacrylamide gel electrophoresis.

Identification of the virus-specific proteins of respiratory syncytial virus temperature-sensitive mutants by immunoprecipitation.

The proteins of Long strain RSV and three temperature-sensitive (ts) mutants of the A2 strain were compared by pulse labeling virus-infected cells with [35S]methionine and [3H]glucosamine followed by analysis of the cell lysates by polyacrylamide gel electrophoresis. At the permissive temperature (30 degrees) proteins ranging in molecular weight from 24,000 to 50,000 (VP24, VP27, VP33, VP44) could be identified. Immunoprecipitation of viral lysates by immune rabbit serum demonstrated antigenic similarity with VP27, VP44, VP50, and VP67 in all ts mutants and Long strain RSV. [3H]Glucosamine labeling demonstrated glycoproteins of 90,000 (GP90) and 50,000 (GP50) in Long strain and GP90 in the ts mutants.