A sample purification method for rugged and high-performance DNA sequencing by capillary electrophoresis using replaceable polymer solutions. A. Development of the cleanup protocol

A method for the cleanup of Sanger DNA sequencing reaction products for capillary electrophoresis analysis with replaceable polymer solutions has been developed. A poly(ether sulfone) ultrafiltration membrane pretreated with linear polyacrylamide was first used to remove template DNA from the sequencing samples. Then, gel filtration in a spin column format (two columns per sample) was employed to decrease the concentration of salts below 10 microM in the sample solution. The method was very reproducible and increased the injected amount of the sequencing fragments 10-50-fold compared to traditional cleanup protocols. Using M13mp18 as template, the resulting cleaned-up single DNA sequencing fragments could routinely be separated to more than 1000 bases with a base-calling accuracy of at least 99% for 800 bases. The method is simple and universal and can be easily automated. In the following paper, a systematic study to determine quantitatively the effects of the sample solution components such as high-mobility ions (e.g., chloride and dideoxynucleotides) and template DNA on the injected amount and separation efficiency of the sequencing fragments is presented.     

The new form of the Netherlands patent bulletin De Industriële Eigendom

The official patent bulletin published by the Netherlands Patent Office, which is called De Industriële Eigendom (Industrial Property), used to appear in two separate parts, one dealing with published applications as filed, and the other with applications published after examination and patents granted. Since 1982 these two parts are published together and a third part dealing with European patents granted and valid for The Netherlands was added.Also the timing of the publication and the lay-out of a number of headings in the bulletin was modified and its outward appearance was modernized. The accessibility of the bulletin to foreigners was increased by the addition of explanatory notes in English.After some historic background information, the nature of the modifications is described and some information is given about the experiences gained in effectuating the changes.     

Primary pericardial mesothelioma

This study reports the results of analysis concerning some chemical characteristics of 63 samples of tomato juice and paste. Copper was determined by atomic absorption spectrometry and its average content was 32 ppm. This is not a critical value for human health and results below the upper limit provided by law of several european and not european countries.     

Hormone replacement therapy and the risk of stroke. Follow-up of a population-based cohort in Sweden

BACKGROUND: The protective effect of postmenopausal estrogen replacement therapy on coronary heart disease has been shown in several studies. However, the effect on stroke is more controversial, and data on estrogen-progestin combinations are sparse. METHODS: A total of 23,088 women living in the Uppsala (Sweden) Health Care Region were identified from pharmacy records as having been prescribed noncontraceptive estrogens during 1977 through 1980. They were followed up from 1977 to 1983 for admissions to the hospital because of a first stroke (International Classification of Diseases, Eighth Revision, codes 430 through 438 and 344). The mean observation time was 5.8 years. The expected number was based on person-years in the cohort and incidence rates in the population of the region. RESULTS: Overall, 361 cases of first stroke were observed in the cohort, as compared with 403.2 expected (relative risk [RR], 0.90; 95% confidence limits, 0.81, 0.99). The RR for acute stroke (International Classification of Diseases, Eighth Revision, codes 431 through 436) was 0.85 (0.75, 0.97). In women younger than 60 years at entry who were prescribed estradiol compounds (1 to 2 mg) or conjugated equine estrogens (0.625 to 1.25 mg), the risk of any stroke was reduced by almost 30% (RR, 0.72; 0.58, 0.88) and the risk of acute stroke was reduced by 40% (RR, 0.61; 0.46, 0.79). Women prescribed a combined estradiol-levonorgestrel brand also had a lowered risk of stroke (RR, 0.61; 0.40, 0.88). Weak compounds (mainly estriol) showed no stroke-protective effect, nor was there any relationship between hormone replacement and risk of subarachnoid hemorrhage. CONCLUSION: Hormone replacement therapy with potent estrogens alone or cyclically combined with progestins can, particularly when started shortly after menopause, reduce the risk of stroke.     

Physiological response of Lactobacillus plantarum to salt and nonelectrolyte stress

In this report, we compared the effects on the growth of Lactobacillus plantarum of raising the medium molarity by high concentrations of KCl or NaCl and iso-osmotic concentrations of nonionic compounds. Analysis of cellular extracts for organic constituents by nuclear magnetic resonance spectroscopy showed that salt-stressed cells do not contain detectable amounts of organic osmolytes, whereas sugar-stressed cells contain sugar (and some sugar-derived) compounds. The cytoplasmic concentrations of lactose and sucrose in growing cells are always similar to the concentrations in the medium. By using the activity of the glycine betaine transport system as a measure of hyperosmotic conditions, we show that, in contrast to KCl and NaCl, high concentrations of sugars (lactose or sucrose) impose only a transient osmotic stress because external and internal sugars equilibrate after some time. Analysis of lactose (and sucrose) uptake also indicates that the corresponding transport systems are neither significantly induced nor activated directly by hyperosmotic conditions. The systems operate by facilitated diffusion and have very high apparent affinity constants for transport (>50 mM for lactose), which explains why low sugar concentrations do not protect against hyperosmotic conditions. We conclude that the more severe growth inhibition by salt stress than by equiosmolal concentrations of sugars reflects the inability of the cells to accumulate K+ (or Na+) to levels high enough to restore turgor as well as deleterious effects of the electrolytes intracellularly.     

Solid state 13C-NMR of cellulose. A relaxation study

Summary The ¹³C spin-lattice relaxation times T₁ of hydrolysed cotton cellulose have been determined in the solid state. T₁ is between 202 and 266 seconds in the crystalline regions of the sample for all the carbons in the anhydroglucose unit and less than 15 seconds in the amorphous part. Based on the relaxation data, spectral assignment of the C2-C3-C5 region is proposed.     

Thermal conductivity and Lorenz number of high-purity praseodymium

These are the first thermal conductivity and Lorenz number measurements to be reported for praseodymium over the temperature range 2–40K. The measurements made on a well-annealed dhcp polycrystalline specimen reveal behavior which is more typical of free-electron-like metals than of rare-earth metals. However, before the specimen was adequately annealed the Lorenz numbers were abnormally high in a manner which resembled those published for many other rare earths.Research supported by the National Research Council of Canada and Artetech International of France.     

Sediment dilution method to determine sorption coefficients of hydrophobic organic chemicals

Sorption coefficients of hydrophobic organic chemicals (HOC) to sediments and soils can easily be underestimated in traditional batch experiments, especially because analysis of the aqueous concentration often includes compounds sorbed to colloidal organic matter. In this work, a "sediment dilution approach" has been combined with measurements of freely dissolved concentrations to determine sorption coefficients of five chlorobenzenes and two chloroanilines in spiked sediment and of two unknown chemicals in field-contaminated sediment. A range of sediment suspensions with different sediment-water ratios was made. Freely dissolved concentrations in these suspensions were measured by negligible depletion solid-phase microextraction (nd-SPME). Sediment-water sorption coefficients (KD) were derived from the decrease of the freely dissolved concentrations as a function of the "dilution factor" (DF = volume water/mass sediment). The determined sorption coefficients were very similar to literature values. The experimental setup provides sorption coefficients without the need for total extractions, and the negligible depletion SPME technique does not require phase separation. The proposed method might be an alternative for batch equilibrium experiments to determine sorption coefficients.